WO2012170827A2 - Pyrazolopyrimidines and related heterocycles as ck2 inhibitors - Google Patents

Pyrazolopyrimidines and related heterocycles as ck2 inhibitors Download PDF

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WO2012170827A2
WO2012170827A2 PCT/US2012/041567 US2012041567W WO2012170827A2 WO 2012170827 A2 WO2012170827 A2 WO 2012170827A2 US 2012041567 W US2012041567 W US 2012041567W WO 2012170827 A2 WO2012170827 A2 WO 2012170827A2
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optionally substituted
alkyl
compound
aryl
heteroaryl
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PCT/US2012/041567
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French (fr)
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WO2012170827A3 (en
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Mustapha Haddach
Joe A. Tran
Fabrice Pierre
Collin F. Regan
Nicholas B. Raffaele
Suchitra Ravula
David M. Ryckman
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Cylene Pharmaceuticals, Inc.
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Publication of WO2012170827A2 publication Critical patent/WO2012170827A2/en
Publication of WO2012170827A3 publication Critical patent/WO2012170827A3/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/53—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with three nitrogens as the only ring hetero atoms, e.g. chlorazanil, melamine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca

Definitions

  • the invention relates in part to molecules having certain biological activities that include, but are not limited to, inhibiting cell proliferation, and modulating certain protein kinase activities.
  • Molecules of the invention modulate, e.g. , Protein Kinase CK2 (called CK2 herein) and are useful to treat conditions associated directly or indirectly with CK2 activities, e.g. , cancers, inflammatory conditions, infectious disorders, pain, immunological disorders, a neurodegenerative disorder (such as Alzheimer's disease and Parkinson's disease), etc.
  • the invention also relates in part to methods for using such compounds, and pharmaceutical compositions containing these compounds.
  • Protein kinase CK2 (formerly called Casein kinase II, referred to herein as "CK2") is a ubiquitous and highly conserved protein serine/threonine kinase.
  • the holoenzyme is typically found in tetrameric complexes consisting of two catalytic (alpha and/or alpha') subunits and two regulatory (beta) subunits.
  • CK2 has a number of physiological targets and participates in a complex series of cellular functions including the maintenance of cell viability.
  • the level of CK2 in normal cells is tightly regulated, and it has long been considered to play a role in cell growth and proliferation.
  • Inhibitors of CK2 that are useful for treating certain types of cancers are described in PCT/US2007/077464, PCT/US2008/074820, PCT/US2009/35609.
  • CK2 is an ancient enzyme on the evolutionary scale; its longevity may explain why it has become important in so many biochemical processes, and why CK2 from hosts have even been co-opted by infectious pathogens (e.g., viruses, protozoa) as an integral part of their survival and life cycle biochemical systems. These same characteristics explain why inhibitors of CK2 are believed to be useful in a variety of medical treatments as discussed herein. Because CK2 is central to many biological processes, as summarized by Guerra & Issinger, Curr. Med. Chem. , 2008, 15: 1870-1886, inhibitors of CK2, including the compounds described herein, should be useful in the treatment of a variety of diseases and disorders.
  • infectious pathogens e.g., viruses, protozoa
  • CK2 has been shown to be associated with acute and chronic myelogenous leukemia, lymphoma and multiple myeloma.
  • enhanced CK2 activity has been observed in solid tumors of the colon, rectum and breast, squamous cell carcinomas of the lung and of the head and neck (SCCHN), adenocarcinomas of the lung, colon, rectum, kidney, breast, and prostate.
  • elevated C 2 has been found to be highly correlated with aggressiveness of neoplasias, and treatment with a CK2 inhibitor of the invention should thus reduce tendency of benign lesions to advance into malignant ones, or for malignant ones to metastasize.
  • C 2 activity level appears to be generally caused by upregulation or overexpression of the active protein rather than by changes that affect activation levels. Guerra and Issinger postulate this may be due to regulation by aggregation, since activity levels do not correlate well with mRNA levels.
  • Excessive activity of CK2 has been shown in many cancers, including SCCHN tumors, lung tumors, breast tumors, and others. Id. Elevated CK2 activity in colorectal carcinomas was shown to correlate with increased malignancy. Aberrant expression and activity of CK2 have been reported to promote increase nuclear levels of NF-kappaB in breast cancer cells.
  • C 2 activity is markedly increased in patients with AML and CML during blast crisis, indicating that an inhibitor of C 2 should be particularly effective in these conditions.
  • Multiple myeloma cell survival has been shown to rely on high activity of CK2, and inhibitors of CK2 were cytotoxic to MM cells.
  • CK2 correlates with efficacy against tumor cells.
  • a CK2 inhibitor inhibited growth of murine pi 90 lymphoma cells. Its interaction with Bcr/Abl has been reported to play an important role in proliferation of Bcr/Abl expressing cells, indicating inhibitors of C 2 may be useful in treatment of Bcr/Abl- positive leukemias.
  • Inhibitors of C 2 have been shown to inhibit progression of skin papillomas, prostate and breast cancer xenografts in mice, and to prolong survival of transgenic mice that express prostate-promoters. Id.
  • C 2 The role of C 2 in various non-cancer disease processes has been recently reviewed. See Guerra & Issinger, Curr. Med. Chem. , 2008, 15: 1870-1886.
  • CK2 is involved in critical diseases of the central nervous system, including, for example, Alzheimer's disease, Parkinson's disease, and rare neurodegenerative disorders such as Guam- Parkinson dementia, chromosome 18 deletion syndrome, progressive supranuclear palsy, Kuf s disease, or Pick's disease. It is suggested that selective CK2-mediated phosphorylation of tau proteins may be involved in progressive neurodegeneration of Alzheimer's disease.
  • recent studies suggest that CK2 plays a role in memory impairment and brain ischemia, the latter effect apparently being mediated by C 2's regulatory effect on the PI3K survival pathways.
  • C 2 has also been shown to be involved in the modulation of inflammatory disorders, for example, acute or chronic inflammatory pain, glomerulonephritis, and autoimmune diseases, including, e.g., multiple sclerosis (MS), systemic lupus erythematosus, rheumatoid arthritis, and juvenile arthritis. It positively regulates the function of the serotonin 5-HT3 receptor channel, activates heme oxygenase type 2, and enhances the activity of neuronal nitric oxide synthase. A selective CK2 inhibitor was reported to strongly reduce pain response of mice when
  • CK2 a nuclear DNA-binding protein
  • Protein kinase C 2 has also been shown to play a role in disorders of the vascular system, such as, e.g., atherosclerosis, laminar shear stress, and hypoxia.
  • CK2 has also been shown to play a role in disorders of skeletal muscle and bone tissue, such as cardiomyocyte hypertrophy, impaired insulin signaling and bone tissue mineralization. In one study, inhibitors of CK2 were effective at slowing angiogenesis induced by growth factor in cultured cells.
  • CK2 inhibitor combined with octreotide (a somatostatin analog) reduced neovascular tufts; thus, the CK2 inhibitors described herein would be effective in combination with a somatostatin analog to treat retinopathy.
  • CK2 has also been shown to phosphorylate GSK, troponin and myosin light chain; thus, C 2 is important in skeletal muscle and bone tissue physiology, and is linked to diseases affecting muscle tissue.
  • CK2 is also involved in the development and life cycle regulation of protozoal parasites, such as, for example, Theileria parva, Trypanosoma cruzi, Leishmania donovani, Herpetomonas muscarum muscarum, Plasmodium falciparum, Trypanosoma brucei, Toxoplasma gondii and Schistosoma mansoni. Numerous studies have confirmed the role of CK.2 in regulation of cellular motility of protozoan parasites, essential to invasion of host cells. Activation of CK2 or excessive activity of CK2 has been shown to occur in hosts infected with Leishmania donovani, Herpetomonas muscarum muscarum, Plasmodium falciparum,
  • C 2 has also been shown to interact with and/or phosphorylate viral proteins associated with human immunodeficiency virus type 1 (HIV-1 ), human papilloma virus, and herpes simplex virus, in addition to other virus types (e.g. , human cytomegalovirus, hepatitis C and B viruses, Borna disease virus, adenovirus, coxsackievirus, coronavirus, influenza, and varicella zoster virus).
  • C 2 phosphorylates and activates HIV- 1 reverse transcriptase and proteases in vitro and in vivo, and promotes pathogenicity of simian-human immunodeficiency virus (SHIV), a model for HIV.
  • HIV human immunodeficiency virus
  • Inhibitors of CK2 are thus able to reduce pathogenic effects of a model of HIV infection.
  • C 2 also phosphorylates numerous proteins in herpes simplex virus and numerous other viruses, and some evidence suggests viruses have adopted CK2 as a phosphorylating enzyme for their essential life cycle proteins. Inhibition of CK2 is thus expected to deter infection and progression of viral infections, which rely upon the host's CK2 for their own life cycles.
  • CK2 is unusual in the diversity of biological processes that it affects, and it differs from most kinases in other ways as well: it is constitutively active, it can use ATP or GTP, and it is elevated in most tumors and rapidly proliferating tissues.
  • CK2's unique structural features enable discovery of highly CK2- specific inhibitors. For all of these reasons, CK2 is a particularly interesting target for drug development, and the invention provides highly effective inhibitors of C 2 that are useful in treating a variety of different diseases and disorders mediated by or associated with excessive, aberrant or undesired levels of CK2 activity.
  • the current invention provides novel compounds of Formula (I), (II), (Ila), (lib), (lie), (lid), (III), (Ilia), (Illb), (IV), (V), (VI), and (Via) including any compound species thereof, as well as the salt, solvate, and/or prodrug thereof, and pharmaceutical compositions containing these compounds or their salt, solvate, and/or prodrugs thereof.
  • the compounds of the present invention show surprisingly greater activity on CK2 and reduced Pirn activity, and thus are advantageously used to treat conditions sensitive to C 2 inhibition such as those described herein.
  • the present compounds are therefore useful to treat conditions mediated by or associated with excessive activity of CK2, with reduced likelihood of off-target effects caused by inhibition of other kinases.
  • the present invention in part provides chemical compounds having certain biological activities that include, but are not limited to, inhibiting cell proliferation, inhibiting angiogenesis, and modulating protein kinase activities.
  • These molecules such as compounds of Formula (I), (II), (Ila), (lib), (lie), (lid), (III), (Ilia), (Illb), (IV), (V), (VI), or (Via) including any compound species thereof, modulate protein kinase CK2 (CK2) and/or PIM activity, and are typically more selective for CK2 activity over other kinases.
  • compositions comprising these molecules in combination with other materials, including other therapeutic agents, and methods for using such compositions.
  • compositions comprising a compound of the present invention as described herein and at least one pharmaceutically acceptable carrier or excipient, or two or more pharmaceutically acceptable carriers and/or excipients.
  • compositions comprising at least one of these compounds can be utilized in methods of treatment such as those described herein.
  • the proteins of the present invention as described herein bind to and inhibit certain kinase proteins, which is believed to be the basis for their pharmaceutical activity.
  • the protein is a C 2 protein, such as a CK2 protein comprising the amino acid sequence of SEQ ID NO: l , 2 or 3 or a substantially identical variant thereof, for example.
  • Substantially identical variants of these include proteins having at least 90% sequence homology with one of these, in one embodiment at least 90% sequence identity; and having at least 50% of the level of in vitro kinase activity of the specified sequence under typical assay conditions.
  • the invention includes methods to modulate the activity of CK2 protein, either in vitro or ex vivo. Suitable methods comprise contacting a system comprising the protein with a compound described herein in an amount effective for modulating the activity of the protein. In certain embodiments the activity of the protein is inhibited, and sometimes the protein is a CK2 protein comprising the amino acid sequence of SEQ ID NO: l , SEQ ID NO:2 or SEQ ID NO:3 or a substantially identical variant thereof, for example. In certain embodiments the C 2 is in a cell or tissue; in other embodiments, it can be in a cell-free system.
  • the cells sometimes are in a cell line, such as a cancer cell line (e.g., breast cancer, prostate cancer, pancreatic cancer, lung cancer, hemopoietic cancer, colorectal cancer, skin cancer, ovary cancer cell line), for example.
  • the cancer cell line is a breast cancer, prostate cancer or pancreatic cancer cell line.
  • the cells sometimes are in a tissue, can be in a subject, at times are in a tumor, and sometimes are in a tumor in a subject.
  • the method further comprises inducing cell apoptosis. Cells sometimes are from a subject having macular degeneration.
  • the cell proliferative condition is a tumor-associated cancer, e.g., a solid or circulating tumor.
  • the cancer sometimes is cancer of the breast, prostate, pancreas, lung, colorectum, skin, or ovary.
  • the cell proliferative condition is a non-tumor cancer, such as a
  • hematopoietic cancer for example, including leukemias, e.g., multiple myeloma and lymphomas.
  • leukemias e.g., multiple myeloma and lymphomas.
  • the cell proliferative condition is macular degeneration in some embodiments.
  • the invention also includes methods for treating cancer or an inflammatory disorder or other disorders described herein that are mediated by excessive activity of one or more of these kinases, in a subject in need of such treatment, comprising: administering to the subject a therapeutically effective amount of a therapeutic agent useful for treating such disorder; and administering to the subject a molecule described herein, e.g., a compound inhibits CK2 in an amount that is effective to enhance a desired effect of the therapeutic agent.
  • the molecule that inhibits CK2 is a compound of Formula (I), (II), (Ila), (lib), (lie), (lid), (III), (Ilia), (Illb), (IV), (V), (VI), (Via), or any compound species thereof, or a pharmaceutically acceptable salt, solvate, and/or prodrug thereof.
  • the desired effect of the therapeutic agent that is enhanced by the molecule that inhibits CK2 is an increase in apoptosis in at least one type of cell.
  • the cell is a cancer cell and the compound is a compound of Formula (I), (II), (Ila), (lib), (lie), (lid), (III), (Ilia), (Illb), (IV), (V), (VI), (Via), or any compound species thereof, that is a potent inhibitor (IC-50 less than about 100 nM, for example) of C 2.
  • the compound has an IC-50 on Pim of less than about 30 nM, and is selective for CK2 over Pim kinases.
  • the IC-50 for inhibition of C 2 is lower by at least a factor of ten than activity on Pim; in preferred embodiments, the compound has an IC-50 for CK2 that is lower than its IC-50 for at least one of Pim-1 , Pim-2 and Pim-3 by about 100-fold or more.
  • the therapeutic agent and the molecule that inhibits C 2 are administered at substantially the same time.
  • the therapeutic agent and molecule that inhibits C 2 sometimes are used concurrently by the subject.
  • the therapeutic agent and the molecule that inhibits C 2 can be combined into one pharmaceutical composition in certain embodiments; in other embodiments they are admistered as separate compositions.
  • compositions of matter comprising a compound described herein and an isolated protein.
  • the protein sometimes is a C 2 protein, such as a C 2 protein comprising the amino acid sequence of SEQ ID NO: l, SEQ ID NO:? nr SEQ ID NO:3 or a substantially identical variant thereof, for example.
  • the protein is a Pirn protein.
  • compositions comprise a compound described herein in combination with a cell.
  • the cell may be from a cell line, such as a cancer cell line.
  • the cancer cell line is sometimes a breast cancer, prostate cancer, pancreatic cancer, lung cancer, hematopoietic cancer, colorectal cancer, skin cancer, or ovary cancer cell line.
  • Compounds of the present invention exert biological activities that include, but are not limited to, inhibiting cell proliferation, reducing angiogenesis, preventing or reducing inflammatory responses and pain, and modulating certain immune responses. Such compounds modulate CK2 activity, as demonstrated by the data herein. Such compounds therefore can be utilized in multiple applications by a person of ordinary skill in the art.
  • compounds described herein can be used, for example, for (i) modulation of protein kinase activity (e.g., CK2 activity), (ii) modulation of cell proliferation, (Hi) modulation of apoptosis, and/or (iv) treatments of cell proliferation related disorders (e.g. , administration alone or co-administration with another molecule).
  • the compounds of Formula (I), (II), (Ha), (lib), (lie), (lid), (III), (Ilia), (Illb), (IV), (V), (VI), or (Via) including any compound species thereof can be used to modulate CK2 activity, in vitro or in vivo, and to treat disorders associated with excessive or undesirable levels of CK2 activity, including cancers, certain inflammatory disorders, vascular disorders, certain skeletal and muscle disorders, and infections such as protozoal parasite infections and some viral infections.
  • compound(s) of the invention refer to compounds encompassed by structural formulae disclosed herein, e.g., Formula (I), (II), (Ila), (lib), (lie), (lid), (III), (Ilia), (Illb), (IV), (V), (VI), and (Via), and include any specific compounds within these formulae whose structure is disclosed herein.
  • Compounds may be identified either by their chemical structure and/or chemical name. When the chemical structure and chemical name conflict, the chemical structure is determinative of the identity of the compound.
  • the present compounds can modulate, i.e., inhibit or enhance, the biological activity of a C 2 protein, a Pim protein or both, and thereby is also referred to herein as a "modulator(s)" or "CK2 and/or Pim modulator(s)".
  • modulator(s) or "CK2 and/or Pim modulator(s)”.
  • Compounds of Formula (I), (II), (Ila), (lib), (lie), (lid), (III), (Ilia), (Illb), (IV), (V), (VI), and (Via), including any specific compounds, i.e., species, described herein are exemplary "modulators".
  • the compounds described herein may contain one or more chiral centers and/or double bonds and therefore, may exist as stereoisomers, such as double-bond isomers (i. e. , geometric isomers such as E and Z), enantiomers or diastereomers.
  • stereoisomers such as double-bond isomers (i. e. , geometric isomers such as E and Z), enantiomers or diastereomers.
  • the invention includes each of the isolated stereoisomeric forms (such as the enantiomerically pure isomers, the E and Z isomers, etc.) as well as mixtures of stereoisomers in varying degrees of chiral purity or percetange of E and Z, including racemic mixtures, mixtures of diastereomers, and mixtures of E and Z isomers.
  • the chemical structures depicted herein encompass all possible enantiomers and stereoisomers of the illustrated compounds including the stereoisomerically pure form (e.g. , geometrically pure, enantiomerically pure or diastereomerically pure) and enantiomeric and stereoisomeric mixtures.
  • Enantiomeric and stereoisomeric mixtures can be resolved into their component enantiomers or stereoisomers using separation techniques or chiral synthesis techniques well known to the skilled artisan.
  • the invention includes each of the isolated stereoisomeric forms as well as mixtures of stereoisomers in varying degrees of chiral purity, including racemic mixtures. It also encompasses the various diastereomers.
  • the compounds may also exist in several tautomeric forms, and the depiction herein of one tautomer is for convenience only, and is also understood to encompass other tautomers of the form shown. Accordingly, the chemical structures depicted herein encompass all possible tautomeric forms of the illustrated compounds.
  • tautomer refers to isomers that change into one another with great ease so that they can exist together in equilibrium: For example, ketone and enol are two tautomeric forms of one compound.
  • a substituted 1 ,2,4-triazole derivative may exist in at least three tautomeric forms as shown below:
  • R T1 is H or optionally substituted alkyl
  • R T2 is an optionally substituted aryl.
  • structures depicted herein are also meant to include compounds which differ only in the presence of one or more isotopically enriched atoms.
  • compounds having the present structures except for the replacement of a hydrogen by a deuterium or tritium, or the replacement of a carbon by 13 C- or 1 C-enriched carbon are within the scope of this invention.
  • the compounds of the present invention may also contain unnatural proportions of atomic isotopes at one or more of atoms that constitute such compounds.
  • the compounds may be radiolabeled with radioactive isotopes, such as for example tritium ( 3 H), iodine-125 ( l25 I) or carbon- 14 ( 14 C). All isotopic variations of the compounds of the present invention, whether radioactive or not, are encompassed within the scope of the present invention.
  • the compounds of the invention often have ionizable groups so as to be capable of preparation as salts.
  • a pharmaceutically acceptable salt may also be used.
  • These salts may be acid addition salts involving inorganic or organic acids or the salts may, in the case of acidic forms of the compounds of the invention be prepared from inorganic or organic bases.
  • the compounds are prepared or used as pharmaceutically acceptable salts prepared as addition products of pharmaceutically acceptable acids or bases.
  • Suitable pharmaceutically acceptable acids and bases are well-known in the art, such as hydrochloric, sulphuric, hydrobromic, acetic, lactic, citric, or tartaric acids for forming acid addition salts, and potassium hydroxide, sodium hydroxide, ammonium hydroxide, caffeine, various amines, and the like for forming basic salts. Methods for preparation of the appropriate salts are well-established in the art.
  • the compounds may contain both an acidic and a basic functional group, in which case they may have two ionized groups and yet have no net charge.
  • solute means a compound formed by solvation (the combination of solvent molecules with molecules or ions of the solute), or an aggregate that consists of a solute ion or molecule, i.e., a compound of the invention, with one or more solvent molecules.
  • solvate When water is the solvent, the corresponding solvate is "hydrate". Examples of hydrates include, but are not limited to, hemihydrate, monohydrate, dihydrate, trihydrate, hexahydrate, etc. It should be understood by one of ordinary skill in the art that the pharmaceutically acceptable salt, and/or prodrug of the present compound may also exist in a solvate form.
  • the solvate is typically formed via hydration which is either part of the preparation of the present compound or through natural absorption of moisture by the anhydrous compound of the present invention.
  • ester means any ester of a present compound in which any of the -COOH functions of the molecule is replaced by a -COOR function, in which the R moiety of the ester is any carbon-containing group which forms a stable ester moiety, including but not limited to alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, » ⁇ -vlalkyl, heterocyclyl, heterocyclylalkyl and substituted derivatives thereof.
  • the hydrolysable esters of the present compounds are the compounds whose carboxyls are present in the form of hydrolysable ester groups.
  • esters are pharmaceutically acceptable and can be hydrolyzed to the corresponding carboxyl acid in vivo.
  • These esters may be conventional ones, including lower alkanoyloxyalkyl esters, e.g., pivaloyloxymethyl and 1-pivaloyloxyethyl esters; lower alkoxycarbonylalkyl esters, e.g., methoxycarbonyloxymethyl, 1 -ethoxycarbonyloxyethyl, and 1- isopropylcarbonyloxyethyl esters; lower alkoxymethyl esters, e.g., methoxymethyl esters, lactonyl esters, benzofuran keto esters, thiobenzofuran keto esters; lower alkanoylaminomethyl esters, e.g., acetylaminomethyl esters.
  • lower alkanoyloxyalkyl esters e.g., pivaloyloxymethyl and 1-pivaloyloxye
  • esters can also be used, such as benzyl esters and cyano methyl esters.
  • Other examples of these esters include: (2,2-dimethyl-l- oxypropyloxy)methyl esters; (lRS)-l-acetoxy ethyl esters, 2-[(2-methylpropyloxy)carbonyl]-2- pentenyl esters, l-[[(l-methylethoxy)carbonyl]- oxy]ethyl esters; isopropyloxycarbonyloxyethyl esters, (5-methy l-2-oxo-l,3- dioxole-4-yl) methyl esters, l-[[(cyclohexyloxy)carbonyl]oxy]ethyl esters; 3,3-dimethyl-2-oxobutyI esters.
  • esters of the compounds of the present invention can be formed at free carboxyls of said compounds by using conventional methods.
  • Representative esters include pivaloyloxymethyl esters, isopropyloxycarbonyloxyethyl esters and (5-methyl-2-oxo-l ,3-dioxole-4-yl)methyl esters.
  • prodrug refers to a precursor of a pharmaceutically active compound wherein the precursor itself may or may not be pharmaceutically active but, upon administration, will be converted, either metabolically or otherwise, into the pharmaceutically active compound or drug of interest.
  • a prodrug can be an ester, ether, or amide form of a pharmaceutically active compound.
  • Various types of prodrugs have been prepared and disclosed for a variety of pharmaceuticals. See, for example, Bundgaard, H. and Moss, J., J. Pharm. Sci. 78: 122-126 (1989). Thus, one of ordinary skill in the art knows how to prepare these prodrugs with commonly employed techniques of organic synthesis.
  • Protecting group refers to a grouping of atoms that when attached to a reactive functional group in a molecule masks, reduces or prevents reactivity of the functional group. Examples of protecting groups can be found in Greene & Wuts, "Protective Groups in Organic Synthesis,” Wiley Interscience, 1999 and Harrison et al., “Compendium of Synthetic Organic Methods", Vols. 1 -8 (John Wiley and Sons, 1971-1996).
  • Representative amino protecting groups include, but are not limited to, formyl, acetyl, trifl"r>roacetyl, benzyl, benzyloxycarbonyl ("CBZ”), tert-butoxycarbonyl (“Boc”), trimethylsilyl (“TMS”), 2-trimethylsilyl-ethanesulfonyl (“SES”), trityl and substituted trityl groups, allyloxycarbonyl, 9-fluorenylmethyloxycarbonyl (“FMOC”), nitro-veratryloxycarbonyl (“NVOC”) and the like.
  • hydroxy protecting groups include, but are not limited to, those where the hydroxy group is either acylated or alkylated such as benzyl, and trityl ethers as well as alkyl ethers, tetrahydropyranyl ethers, trialkylsilyl ethers and allyl ethers.
  • pharmaceutically acceptable means suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended use within the scope of sound medical judgment.
  • Excipient refers to a diluent, adjuvant, vehicle, or carrier with which a compound is administered.
  • an “effective amount” or “therapeutically effective amount” is the quantity of the present compound in which a beneficial outcome is achieved when the compound is administered to a patient or alternatively, the quantity of compound that possesses a desired activity in vivo or in vitro.
  • a beneficial clinical outcome includes reduction in the extent or severity of the symptoms associated with the disease or disorder and/or an increase in the longevity and/or quality of life of the patient compared with the absence of the treatment.
  • a "beneficial clinical outcome” includes a reduction in tumor mass, a reduction in the rate of tumor growth, a reduction in metastasis, a reduction in the severity of the symptoms associated with the cancer and/or an increase in the longevity of the subject compared with the absence of the treatment.
  • the precise amount of compound administered to a subject will depend on the type and severity of the disease or condition and on the characteristics of the patient, such as general health, age, sex, body weight and tolerance to drugs. It will also depend on the degree, severity and type of proliferative disorder. The skilled artisan will be able to determine appropriate dosages depending on these and other factors.
  • alkyl straight-chain, branched-chain and cyclic monovalent hydrocarbyl radicals, and combinations of these, which contain only C and H when they are unsubstituted. Examples include methyl, ethyl, isobutyl, cyclohexyl, cyclopentylethyl, 2-propenyl, 3-butynyl, and the like.
  • the total number of carbon atoms in each such group is sometimes described herein, e. v., when the group can contain up to ten carbon atoms it can be represented as.l-lOC or as C1-C10 or Cl-10.
  • heteroatoms N, O and S typically
  • the numbers describing the group though still written as e.g., C1-C6, represent the sum of the number of carbon atoms in the group plus the number of such heteroatoms that are included as replacements for carbon atoms in the backbone of the ring or chain being described.
  • a ring is included, it is understood that the group contains at least three carbon atoms as a 3- membered ring is the smallest size for a ring.
  • the alkyl, alkenyl and alkynyl substituents of the invention contain 1 -lOC (alkyl) or 2-10C (alkenyl or alkynyl), or 3- IOC when a ring is included. In one embodiment, they contain 1-8C (alkyl) or 2-8C (alkenyl or alkynyl) or 3-8C when a ring is included.
  • a single group can include more than one type of multiple bond, or more than one multiple bond; such groups are included within the definition of the term "alkenyl” when they contain at least one carbon-carbon double bond, and are included within the term "alkynyl” when they contain at least one carbon-carbon triple bond; provided, however, that the presence of multiple bonds does not produce an aromatic ring.
  • Alkyl, alkenyl and alkynyl groups are often optionally substituted to the extent that such substitution makes sense chemically.
  • Substituted when used to modify a specified group or radical, means that one or more hydrogen atoms of the specified group or radical are each, independently of one another, replaced with the same or different substituent(s).
  • each R b is independently hydrogen or R a ; and each R c is independently R b or alternatively, the two R°s may be taken together with the nitrogen atom to which they are bonded form a 4-, 5-, 6- or
  • cycloheteroalkyl which may optionally include from 1 to 4 of the same or different additional heteroatoms selected from the group consisting of O, N and S.
  • -NR C R C is meant to include -NH 2 , -NH-alkyl, N-pyrrolidinyl and N-morpholinyl.
  • a substituted alkyl is meant to include -alkylene-O-alkyl, -alkylene-heteroaryl, -alkylene-cycloheteroalkyl, -alkylene-C(0)OR b , -alkylene-C(0)NR b R b , and -CH 2 -CH 2 -C(0)- C3 ⁇ 4.
  • the one or more substituent groups, taken together with the atoms to which they are bonded, may form a cyclic ring including cycloalkyl and cycloheteroalkyl.
  • substituent groups useful for substituting unsaturated carbon atoms in the specified group or radical include, but are not limited to, -R a , halo, -O " , -OR b , -SR b , -S " , -NR C R C , trihalomethyl, -CF 3 , -CN, -OCN, -SCN, -NO, -N0 2 , -N 3 , -S(0) 2 R b , -S(0) 2 0 ⁇ -S(0) 2 OR b , -OS(0) 2 R b , -OS(0) 2 0 " , -OS(0) 2 OR b , -P(0)(Cr) 2!
  • cycloheteroalkyl groups include, but are not limited to, -R a , -O " , -OR b , -SR b , -S " , -NR C R C , trihalomethyl, -CF 3 , -CN, -NO, -N0 2 , -S(0) 2 R b , -S(0) 2 0-, -S(0) 2 OR b , -OS(0) 2 R b , -OS(0) 2 0 ⁇ -OS(0) 2 OR b , -P(0)(0 " ) 2 , -P(0)(OR b )(0-), -P(0)(OR b )(OR b ), -C(0)R b , -C(S)R b , -C(NR b )R b , -C(0)OR b , -C(0)NR c R c , -C(NR b
  • R a of -C ⁇ C-R a is H or Me.
  • Heteroalkyl “heteroalkenyl”, and “heteroalkynyl” and the like are defined similarly to the corresponding hydrocarbyl (alkyl, alkenyl and alkynyl) groups, but the 'hetero' terms refer to groups that contain 1 -3 O, S or N heteroatoms or combinations thereof within the backbone residue; thus at least one carbon atom of a corresponding alkyl, alkenyl, or alkynyl group is replaced by one of the specified heteroatoms to form, respectively, a heteroalkyl, heteroalkenyl, or heteroalkynyl group.
  • heteroforms of alkyl, alkenyl and alkynyl groups are generally the same as for the corresponding hydrocarbyl groups, and the substituents that may be present on the heteroforms are the same as those described above for the hydrocarbyl groups.
  • substituents that may be present on the heteroforms are the same as those described above for the hydrocarbyl groups.
  • such groups do not include more than two contiguous heteroatoms except where an oxo group is present on N or S as in a nitro or sulfonyl group.
  • alkyl as used herein includes cycloalkyl and cycloalkylalkyl groups
  • cycloalkyl may be used herein to describe a carbocyclic non-aromatic group that is connected via a ring carbon atom
  • cycloalkylalkyl may be used to describe a carbocyclic non- aromatic group that is connected to the molecule through alkyl linker.
  • heterocyclyl may be used to describe a non-aromatic cyclic group that contains at least one heteroatom (typically selected from N, O and S) as a ring member and that is connected to the molecule via a ring atom, which may be C (carbon-linked) or N (nitrogen- linked); and “heterocyclylalkyl” may be used to describe such a group that is connected to another molecule through a linker.
  • the heterocyclyl can be fully saturated or partially saturated, but non-aromatic.
  • the sizes and substituents that are suitable for the cycloalkyl, cycloalkylalkyl, heterocyclyl, and heterocyclylalkyl groups are the same as those described above for alkyl groups.
  • the heterocyclyl groups typically contain 1 , 2 or 3 heteroatoms, selected from N, O and S as ring members; and the N or S can be substituted with the groups commonly found on these atoms in heterocyclic systems. As used herein, these terms also include rings that contain a double bond or two, as long as the ring that is attached is not aromatic.
  • the substituted cycloalkyl and heterocyclyl groups also include cycloalkyl or heterocyclic rings fused to an aromatic ring or heteroaromatic ring, provided the point of attachment of the group is to the cycloalkyl or heterocyclyl ring rather than to the aromatic / heteroaromatic ring.
  • cycloalkyl and heterocyclyl groups described herein can be substituted to the extent permitted by their valence and stability considerations, which are well understood by those of skill in the art.
  • Substituents for the cycloalkyl and heterocyclyl rings or ring systems include those described herein as suitable for placement on alkyl groups.
  • acyl encompasses groups comprising an alkyl, alkenyl, alkynyl, aryl or arylalkyl radical attached at one of the two available valence positions of a carbonyl carbon atom
  • heteroacyl refers to the corresponding groups wherein at least one carbon other than the carbonyl carbon has been replaced by a heteroatom chosen from N, O and S.
  • Acyl and heteroacyl groups are bonded to any group or molecule to which they are attached through the open valence of the carbonyl carbon atom. Typically, they are C1 -C8 acyl groups, which include formyl, acetyl, pivaloyl, and benzoyl, and C2-C8 heteroacyl groups, which include methoxyacetyl, ethoxycarbonyl, and 4-pyridinoyl.
  • the hydrocarbyl groups, aryl groups, and heteroforms of such groups that comprise an acyl or heteroacyl group can be substituted with the substituents described herein as generally suitable substituents for each of the corresponding component of the acyl or heteroacyl group.
  • Aromaatic moiety or aryl moiety refers to a monocyclic or fused bicyclic moiety having the well-known characteristics of aromaticity; examples include phenyl and naphthyl.
  • heteroaryl refers to such monocyclic or fused bicyclic ring systems which contain as ring members one or more heteroatoms selected from O, S and N. The inclusion of a heteroatom permits aromaticity in 5-membered rings as well as 6-membered rings.
  • Typical heteroaromatic systems include monocyclic C5-C6 aromatic groups such as pyridyl, pyrimidyl, pyrazinyl, thienyl, furanyi, pyrrolyi, pyrazoiyl, thiazolyi, oxazoiyl, triazolyl, triazinyl, tetrazolyl, tetrazinyl, and imidazolyl and the fused bicyclic moieties formed by fusing one of these monocyclic groups with a phenyl ring or with any of the heteroaromatic monocyclic groups to form a C8-C 10 bicyclic group such as indolyl, benzimidazolyl, indazolyl, benzotriazolyl, isoquinolyl, quinolyl, benzothiazolyl, benzofuranyl, pyrazolopyridyl, quinazolinyl, quinoxalinyl, cinnolinyl, and
  • Any monocyclic or fused ring bicyclic system which has the characteristics of aromaticity in terms of electron distribution throughout the ring system is included in this definition. It also includes bicyclic groups where at least the ring which is directly attached to the remainder of the molecule has the characteristics of aromaticity.
  • the ring systems typically contain 5-12 ring member atoms and up to four heteroatoms selected from N, O and S.
  • the monocyclic heteroaryls contain 5-6 ring members and up to three such heteroatoms
  • the bicyclic heteroaryls contain 8- 10 ring members and up to four such heteroatoms.
  • the number and placement of heteroatoms in such rings is in accordance with the well-known limitations of aromaticity and stability, where stability requires the
  • heteroaromatic group to be stable enough to be exposed to water without rapid degradation.
  • an arylalkyl substituent may be substituted on the aryl portion with substituents described herein as typical for aryl groups, and it may be further substituted on the alkyl portion with substituents described herein as typical or suitable for alkyl groups.
  • a substituent group contains two R groups on the same or adjacent atoms (e.g., -NR2, or -NR-C(O)R)
  • the two R groups can optionally be taken together with the atom(s) in the substituent group to which the are attached to form a ring having 5-8 ring members, which can be substituted as allowed for the R itself, and can contain an additional heteroatom (N, O or S) as a ring member.
  • arylalkyl and heteroarylalkyl refer to aromatic and heteroaromatic ring systems which are bonded to their attachment point through a linking group such as an alkylene, including substituted or unsubstituted, saturated or unsaturated, cyclic or acyclic linkers.
  • linker is C1 -C8 alkyl or a hetero form thereof.
  • linkers may also include a carbonyl group, thus making them able to provide substituents as an acyl or heteroacyl moiety.
  • An aryl or heteroaryl ring in an arylalkyl or heteroarylalkyl group may be substituted with the same substituents described above for aryl groups.
  • an arylalkyl group includes a phenyl ring optionally substituted with the groups defined above for aryl groups and a C 1 -C4 alkylene that is unsubstituted or is substituted with one or two C1 -C4 alkyl groups or heteroalkyl groups, where the alkyl or heteroalkyl groups can optionally cyclize to form a ring such as cyclopropane, dioxolane, or oxacyclopentane.
  • a heteroarylalkyl group preferably includes a C5-C6 monocyclic heteroaryl group that is optionally substituted with the groups described above as substituents typical on aryl groups and a C1 -C4 alkylene that is unsubstituted or is substituted with one or two C 1 -C4 alkyl groups or heteroalkyl groups, or it includes an optionally substituted phenyl ring or C5-C6 monocyclic heteroaryl and a C1 -C4 heteroalkylene that is unsubstituted or is substituted with one or two C 1 -C4 alkyl or heteroalkyl groups, where the alkyl or heteroalkyl groups can optionally cyclize to form a ring such as cyclopropane, dioxolane, or oxacyclopentane.
  • substituents may be on either the alkyl or heteroalkyl portion or on the aryl or heteroaryl portion of the group.
  • the substituents optionally present on the a' ⁇ vl or heteroalkyl portion are the same as those described above for alkyl groups generally; the substituents optionally present on the aryl or heteroaryl portion are the same as those described above for aryl groups generally.
  • Arylalkyl groups as used herein are hydrocarbyl groups if they are unsubstituted, and are described by the total number of carbon atoms in the ring and alkylene or similar linker.
  • a benzyl group is a C7-arylalkyl group
  • phenylethyl is a C8-arylalkyl.
  • Heteroarylalkyl refers to a moiety comprising an aryl group that is attached through a linking group, and differs from “arylalkyl” in that at least one ring atom of the aryl moiety or one atom in the linking group is a heteroatom selected from N, O and S.
  • the heteroarylalkyl groups are described herein according to the total number of atoms in the ring and linker combined, and they include aryl groups linked through a heteroalkyl linker; heteroaryl groups linked through a hydrocarbyl linker such as an alkylene; and heteroaryl groups linked through a heteroalkyl linker.
  • C7-heteroarylalkyl would include
  • Alkylene refers to a divalent hydrocarbyl group; because it is divalent, it can link two other groups together. Typically it refers to - ⁇ CH2) n - where n is 1 -8 and preferably n is 1 -4, though where specified, an alkylene can also be substituted by other groups, and can be of other lengths, and the open valences need not be at opposite ends of a chain. Thus -CH(Me)- and -C(Me)2- may also be referred to as alkylenes, as can a cyclic group such as
  • substituents include those typically present on alkyl groups as described herein.
  • any alkyl, alkenyl, alkynyl, acyl, or aryl or arylalkyl group or any heteroform of one of these groups that is contained in a substituent may itself optionally be substituted by additional substituents.
  • the nature of these substituents is similar to those recited with regard to the primary substituents themselves if the substituents are not otherwise described.
  • R" is alkyl
  • this alkyl may optionally be substituted by the remaining substituents listed as embodiments for R x where this makes chemical sense, and where this does not undermine the size limit provided for the alkyl per se; e.g.
  • alkyl substituted by alkyl or by alkenyl would simply extend the upper limit of carbon atoms for these embodiments, and is not included.
  • each such alkyl, alkenyl, alkynyl, acyl, or aryl group may be substituted with a number of substituents according to its available valences; in particular, any of these groups may be substituted with fluorine atoms at any or all of its available valences, for example.
  • Heteroform refers to a derivative of a group such as an alkyl, aryl, or acyl, wherein at least one carbon atom of the designated carbocyclic group has been replaced by a heteroatom selected from N, O and S.
  • the heteroforms of alkyl, alkenyl, alkynyl, acyl, aryl, and arylalkyl are heteroalkyl, heteroalkenyl, heteroalkynyl, heteroacyl, heteroaryl, and heteroarylalkyl, respectively. It is understood that no more than two N, O or S atoms are ordinarily connected sequentially, except where an oxo group is attached to N or S to form a nitro or sulfonyl group.
  • Halo as used herein includes fluoro, chloro, bromo and iodo. Fluoro and chloro are often preferred.
  • Amino refers to NH2, but where an amino is described as “substituted” or “optionally substituted”, the term includes NR'R" wherein each R' and R" is independently H, or is an alkyl, alkenyl, alkynyl, acyl, aryl, or arylalkyl group or a heteroform of one of these groups, and each of the alkyl, alkenyl, alkynyl, acyl, aryl, or arylalkyl groups or heteroforms of one of these groups is optionally substituted with the substituents described herein as suitable for the corresponding group.
  • R' and R" are taken together with the N to which they are attached to form a 3-8 membered ring which may be saturated, unsaturated or aromatic and which contains 1 -3 heteroatoms independently selected from N, O and S as ring members, and which is optionally substituted with the substituents described as suitable for alkyl groups or, if NR'R" is an aromatic group, it is optionally substituted with the substituents described as typical for heteroaryl groups.
  • the term “carbocycle”, “carbocyclyl”, or “carbocyclic” refers to a cyclic ring containing only carbon atoms in the ring, whereas the term “heterocycle” or “heterocyclic” refers to a ring comprising a heteroatom.
  • the carbocyclyl can be fully saturated or partially saturated, but non-aromatic.
  • the carbocyclyl encompasses cycloalkyl.
  • the carbocyclic and heterocyclic structures encompass compounds having monocyclic, bicyclic or multiple ring systems; and such systems may mix aromatic, heterocyclic, and carbocyclic rings.
  • heteroatom refers to any atom that is not carbon or hydrogen, such as nitrogen, oxygen or sulfur. When it is part of the backbone or skeleton of a chain or ring, a heteroatom must be at least divalent, and will typically be selected from N, O, P, and S.
  • heterocycles and heteroaryls include but are not limited to tetrahydrofuran, 1 ,3-dioxolane, 2,3-dihydrofuran, pyran, tetrahydropyran, benzofuran, isobenzofuran, 1 ,3-dihydro-isobenzofuran, isoxazole, 4,5-dihydroisoxazole, piperidine, pyrrolidine,- pyrrolidin-2-one, pyrrole, pyridine, pyrimidine, octahydro-pyrrolo[3,4-Z>]pyridine, piperazine, pyrazine, morpholine, thiomorpholine, imidazole, imidazolidine 2,4-dione, 1 ,3- dihydrobenzimidazol-2-one, indole, thiazole, benzothiazole, thiadiazole, thiophene, tetrahydr
  • the present invention provides compounds having the general formula (1):
  • Z 4 is N or CR 5 ;
  • R 5 is halo, -CN, -R,-OR, (0) n R, -COOR, -CONR 2 , -NR 2 , optionally substituted aryl, or optionally substituted heteroaryl;
  • R 2 , R 3 and R 4 are each independently H or optionally substituted CI -CIO alkyl
  • X is O, S, or NR 6 ;
  • Y is O or S or NR 10 ;
  • R 6 and R 10 are each independently selected from H, CN, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C1-C4 alkoxy, optionally substituted C6-C10 aryl, optionally substituted heteroaryl, and NR 7 R 8 ;
  • Z is O or S
  • L is a covalent bond, -NR 7 -, -0-, -S(0) n -, -(CR 7 R 8 ) m -NR 9 -, -(CR 7 R 8 ) m -0-, or -(CR 7 R 8 ) m -
  • W is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, or optionally substituted carbocyclyl;
  • each R 7 and R 8 and R 9 is independently selected from H, optionally substituted CI -CIO alkyl, optionally substituted heteroalkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl;
  • R 7 and R 8 taken together on a single carbon atom or on adjacent connected carbon atoms of (CR 7 R 8 ) m whether alone or as part of another group, form a 3 to 8 membered carbocyclic ring or heterocyclic ring;
  • R 7 and R 8 taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 to 10 membered heterocyclic or heteroaryl ring that optionally contains one or more additional heteroatom selected from N, O and S as a ring member;
  • each m is independently 1 , 2, 3 or 4;
  • G is NR 1A R IB , OR 1A , or SR 1A ;
  • R 1A is H or optionally substituted C1-C10 alkyl
  • R IB is H, optionally substituted C I -C I O alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, or optionally substituted heteroarylalkyl; each R is independently H or optionally substituted C 1 -C4 alkyl; or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member; and
  • each n is independently 0, 1 , or 2.
  • Z 4 is N or CH.
  • R 3 and R 4 are both H.
  • R 2 is H, -CH 3 , halo, -OCH 3 , or
  • Y is O or S.
  • Z is O.
  • X is selected from the group consisting of NH, O, and S.
  • L is a covalent bond; and W is optionally substituted aryl, optionally substituted heteroaryl, or optionally substituted heterocyclyl.
  • L is -NR 7 -, -0-, -S(0) n -, -(CR 7 R 8 ) m -NR 9 -, -(CR 7 R 8 ) m -0-, or -(CR 7 R 8 ) m -S(0) n -;
  • R 7 and R 8 and R 9 are each independently H or C1 -C 10 alkyl; and W is optionally substituted aryl or optionally substituted carbocyclyl.
  • W-L- is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, (optionally substituted aryl)-NR 7 -, (optionally substituted heteroaryl)-NR 7 -, (optionally substituted aryl)-CH 2 -NR 7 -, (optionally substituted heteroaryl)-CH 2 -NR 7 -, (optionally substituted aryl)-0-, (optionally substituted heteroaryl)-0-, (optionally substituted aryl)-S-, (optionally substituted heteroaryl)-S-, (optionally substituted aryl)-CH 2 -0-, or (optionally substituted heteroaryl)-CH 2 -0-; and R 7 is each independently H or C I -C I O alkyl.
  • Z 4 is N or CR 5 ;
  • R 5 is -R, -CN, -OR, -Ar, COOR, - NR 2 ;
  • Z and Y are both O;
  • R 2 and R 4 are both H;
  • R 3 is H, optionally substituted CI -CI O alkyl, or optionally substituted C1 -C 10 heteroalkyl;
  • X is NH;
  • R IB is optionally substituted
  • L is a covalent bond, -NR 7 -, or -CH 2 -NR 7 -;
  • R 7 is H or C 1 -C4 alkyl; and
  • W is optionally substituted aryl or optionally substituted heteroaryl.
  • the one or more substituents are selected from the group consisting of optionally substituted CI -CI O alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted CI -C I O heteroalkyl, -OR, halo, -S(0)R, -S(0) 2 R, -CN, -NR 2 , -SR, -N0 2 , -C(0)R, - C(0)OR, -NHC(0)R, -NRC(0)R, -C(0)NR 2 , -CF 3 , optionally substituted aryl, and optionally substituted heteroaryl; and each R is independently selected from H and and optionally substituted C 1-C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member
  • the one or more substitutents are selected from the group consisting of phenyl, thiophene, pyridine, pyrimidine, pyrazine, pyridazine, pyrrole, furan, thiofuran, oxazole, isooxazole, thiazole, quinoline, isoquinoline, indole, and pyrazole, wherein each substitutent is optionally substituted with one or more substitutents selected from the group consisting of optionally substituted C1 -C10 alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted CI -C I O heteroalkyl, -OR, halo, -S(0)R, -S(0) 2 R, -CN, -NR 2 , -SR, -N0 2 , -C(0)R, -C(0)OR, -NHC(0)R, -NRC(0)R, -C(0)OR, -NH
  • the one or more substitutents are selected from the group consisting of aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine, tetrahydrofuran, pyran, thiopyran, thiomorpholine, thiomorpholine S-oxide, thiomorpholine S-dioxide, oxazoline, tetrahydrothiophene, piperidine, tetrahydropyran, thiane, imidazolidine, oxazolidine, thiazolidine, dioxolane, dithiolane, piperazine, oxazine, dithiane, morpholine, and dioxane, wherein each substitutent is optionally substituted with one or more substitutents selected from the group consisting of optionally substituted C1 -C10 alkyl, optionally substituted carbocycly
  • the one or more substitutents are selected from the group consisting of CI, Br, F, -OCF3, -CF 3 , -CH3, -C 2 H 5 , -C3H7, -OCH 2 0-, OCH 2 CH 2 0-, -SCH 3 , -OCH3, -NHCOCH3, - NH 2 , -COCH 3 , -N0 2 , -0(CH 2 )i. 6 -COOR, and -S0 2 CH 3 .
  • W can be an optionally substituted phenyl or an optionally substituted pyridine.
  • p 0, 1 , 2 or 3;
  • Z 4 is N or CR 5 ;
  • R 5 is H or C l -C4 alkyl
  • R 3 is H, C1 -C 10 alkyl, or C 1 -C10 heteroalkyl
  • R 7 is H or C l -C4 alkyl
  • Ar is aryl or heteroaryl which is optionally substituted by one or more substituents selected from the group consisting of optionally substituted C I -C I O alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted C2-C 10 alkenyl, optionally substituted C2-C10 alkynyl, -NR 2 ; -OR, S(0) n R; halo, -CN, -N0 2 , -C(0)OR, - C(0)NR 2 , or -C(0)R;
  • n 0, 1 , or 2;
  • each R is independently H or optionally substituted C 1 -C4 alkyl; or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member; and
  • R IB is selected from H, optionally substituted C I -C I O alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylaikyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, or optionally substituted heteroarylalkyl.
  • Ar is optionally substituted phenyl or pyridinyl.
  • p is 0 or 1 .
  • the compound has structural Formula (Ila), (lib), (H
  • Z 4 is N or CR 5 ;
  • R 5 is H or C l -C4 alk l
  • X is NH, or S
  • R 3 is H, C1 -C10 alkyl or Cl -CI O heteroalkyl
  • R 7 is H or C l -C4 alkyl
  • q 0, 1 , 2, 3, 4, or 5;
  • t 0, 1 , 2, 3, or 4;
  • A is optionally substituted C1 -C10 alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted C2-C 10 alkenyl, optionally substituted C2-C10 alkynyl, -NR 2 ; -OR, S(0) n R; halo, -CN, -N0 2 , -C(0)OR, -C(0)NR 2 , or -C(0)R;
  • n 0, 1 , or 2;
  • each R is independently H or optionally substituted C 1 -C4 alkyl; or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member; and
  • R 1B is cyclopropyl or cyclopropylmethylene.
  • A is selected from the group consisting of phenyl, thiophene, pyridine, pyrimidine, pyrazine, pyridazine, pyrrole, furan, thiofuran, oxazole, isooxazole, thiazole, quinoline, isoquinoline, indole, and pyrazole, each of which is optionally substituted with one or more substitutents selected from the group consisting of optionally substituted C 1 -C10 alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted C I -C I O heteroalkyl, -OR, halo, -S(0)R, -S(0) 2 R, -CN, -NR 2 , -SR, -N0 2 , -C(0)R, -C(0)OR, -NHC(0)R, -NRC(0)R,
  • A is selected from the group consisting of aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine,
  • A is selected from the group consisting of CI, Br, F, -OCF3, -CF 3 , -CH 3 , -C 2 H 5 , -C 3 H 7 , -OCH 2 0-, OCH 2 CH 2 0-, -SCH 3 , - OCH 3 , -NHCOCH 3 , -NH 2 , -COCH 3 , -N0 2 , -0(CH 2 ) 1-6 -COOR, and -S0 2 CH 3 .
  • the compound has a structural Formula (III):
  • Z 4 is N or CR 5 ;
  • R 5 is halo, -CN, -R,-OR, -S(0) n R, -COOR, -CONR 2 , -NR 2 , optionally substituted aryl, or optionally substituted heteroaryl;
  • R 2 , R 3 and R 4 are each independently is H or optionally substituted C 1 -C10 alkyl;
  • X is O, S, or R 6 ;
  • Y is O, S, or NR 10 ;
  • R 6 and R 10 are each independently selected from H, CN, optionally substituted C 1 -C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C1 -C4 alkoxy, optionally substituted C6-C 10 aryl, and optionally substituted heteroaryl;
  • Z is O or S
  • hCy is an optionally substituted 5 to 10 membered heterocyclic or heteroaryl ring that contains one or more heteroatom selected from N, O and S as a ring member;
  • G is NR 1A R 1B , OR 1A , or SR 1A ;
  • R ,A is H or optionally substituted C1 -C 10 alkyl
  • R IB is selected from H, optionally substituted C I -CI O alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, or optionally substituted heteroarylalkyl;
  • each R is independently H or optionally substituted C 1 -C4 alkyl; or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member; and
  • n 0, 1 , or 2.
  • hCy is selected from the group consisting of thiophene, pyridine, pyrimidine, pyrazine, pyridazine, pyrrole, furan, thiofuran, oxazole, isooxazole, thiazole, quinoline, isoquinoline, indole, and pyrazole, each of which is optionally substituted with one or more substitutents selected from the group consisting of optionally substituted C I -C I O alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted C I -C I O heteroalkyl, -OR, halo, -S(0)R, -S(0) 2 R, -CN, -NR 2 , -SR, -N0 2 , - C(0)R, -C(0)OR, -NHC(0)R, -NRC(0)R, -C(0)NR 2 , -CF 3
  • the compound has a structural Formula (Ilia):
  • each A is independently optionally substituted C I -CI O alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted C2-C10 alkenyl, optionally substituted C2-C 10 alkynyl, -NR 2 ; -OR, S(0) n R; halo, -CN, -N0 2 , -C(0)OR, - C(0)NR 2 , or -C(0)R;
  • c 0, 1 , 2, or 3;
  • n 0, 1 , or 2;
  • each R is independently H or optionally substituted C 1 -C4 alkyl; or alternatively, the two
  • R groups taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member.
  • Z 4 is CR 5 ; R 2 and R 4 are H; Z and Y are O; X is NH or S; G is NR IA R I B ; R 1A is H or methyl; R 1B is optionally substituted C I -C I O alkyl, optionally substituted CI -CI O cylcoalkyl, optionally substituted heterocyclyl, optionally substituted C 1 -C 10 alkylether, optionally substituted C1 -C 10 alkylaminoalkyl, optionally substituted C I -C I O alkoxyalkyl, optionally substituted heterocyclylalkyl, alkylaryl, aralkyl, alkylheteroaryl, heteroarylalkyl, or optionally substituted aryl.
  • R 1B is cyclopropyl, cyclopropylmethylene, cyclobutyl, cyclohexyl, optionally substituted morpholinylalkyl, or optionally substituted phenyl.
  • A is -CN, C 1 -C4 alkyl, - C(0)OR, or -C(0)NR 2 .
  • hCy is selected from the group consisting of aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine, tetrahydrofuran, pyran, thiopyran, thiomorpholine, thiomorpholine S-oxide, thiomorpholine S-dioxide, oxazoline, tetrahydrothiophene, piperidine, tetrahydropyran, thiane, imidazolidine, oxazolidine, thiazolidine, dioxolane, dithiolane, piperazine, oxazine, dithiane, morpholine, and dioxane, each of which is optionally substituted with one or more substitutents selected from the group consisting of optionally substituted C1 -C10 alkyl, optionally substituted carbocyclyl, optionally substituted
  • the compound has a structural Formula (Illb):
  • D is N or CH; a and b are independently 1 , 2, or 3;
  • Z 4 is 1M or CR 5 ;
  • R 5 is halo, -CN, -R,-OR, -S(0) n R, -COOR, -CONR 2 , -NR 2 , optionally substituted aryl, or optionally substituted heteroaryl;
  • R 2 , R 3 and R 4 are each independently selected from H and optionally substituted CI -C I O alkyl;
  • X is O, S, or NR 6 ;
  • Y is O or S or NR 10 ;
  • Z is O or S
  • R 6 and R 10 are each independently selected from H, CN, optionally substituted C 1 -C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C1 -C4 alkoxy, optionally substituted C6-C 10 aryl, and optionally substituted heteroaryl;
  • L 2 is optionally substituted CI -CIO alkyl, optionally substituted CI -CIO heteroalkyl, optionally substituted C2-C 10 alkenyl, optionally substituted C2-C10 alkynyl, optionally substituted aryl, optionally substituted heteroaryl, -C(O)-, -S(0) n -, or -CR 2 -;
  • E is -OR, -NR, optionally substituted C 1 -C10 alkyl, optionally substituted C1 -C 10 heteroalkyl, optionally substituted aryl, or optionally substituted heteroaryl;
  • R IB is H, optionally substituted C I -C I O alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, or optionally substituted heteroarylalkyl; and
  • each R is independently selected from H and optionally substituted C 1 -C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member; and
  • n 0, 1 , or 2.
  • Z 4 is N or CH; R 2' , R 3 , and R 4 are H; Z and Y are O; X is NH; and R I B is cyclopropyl or cyclopropylmethylene.
  • a and b are hr>th 2.
  • L 2 is -C(O)- or -S(0)2-; and E is optionally substituted aryl or optionally substituted heteroaryl.
  • L 2 is -CH 2 -; and E is optionally substituted C 1 -C4 alkyl, optionally substituted aryl, or optionally substituted heteroaryl.
  • the optionally substituted aryl is optionally substituted phenyl
  • the optionally substituted heteroaryl is optionally substituted pyridyl.
  • the optionally substituted aryl or the optionally substituted heteroaryl is independently optionally substituted by one or more substituents selected from the group consisting of optionally substituted C l -CIO alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted Cl - CI O heteroalkyl, -OR, halo, -S(0)R, -S(0) 2 R, -CN, -NR 2) -SR, -N0 2 , -C(0)R, -C(0)OR, - NHC(0)R, -NRC(0)R, -C(0)NR 2 , optionally substituted aryl, and optionally substituted heteroaryl; and each R is independently selected from H and and optionally substituted C1 -C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic
  • L 1 is O or S
  • L 3 is a covalent bond or an optionally substituted C 1 -C4 alkylene;
  • Z 4 each independently represent N or CR 5 ;
  • R 5 is halo, -CN, -R,-OR, -S(0) n R, -COOR, -CONR 2 , -NR 2 , optionally substituted aryl, or optionally substituted heteroaryl;
  • each R is independently selected from H and optionally substituted C 1 -C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member;
  • n 0, 1 , or 2;
  • R 2 , R 3 and R 4 are each independently selected from H and optionally substituted CI -CI O alkyl;
  • X is O, S, or NR 6 ;
  • Y is O or S or NR 10 ;
  • R 6 and R 10 are each independently selected from H, CN, optionally substituted C 1 -C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C 1 -C4 alkoxy, optionally substituted C6-C 10 aryl, and optionally substituted heteroaryl;
  • Z is O or S
  • Ar is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, or optionally substituted heterocyclyl;
  • R IB is selected from H, optionally substituted C I -CI O alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl.
  • Z 4 is CH; R 2 , R 3 , and R 4 are H; Z and Y are O; X is
  • R 1 is cyclopropyl
  • L 3 is a covalent bond or an optionally substituted C1 -C4 alkylene
  • Ar is optionally substituted aryl or heteroaryl.
  • L 3 is a covalent bond or methylene.
  • Ar is optionally substituted phenyl, optionally substituted pyridyl, or optionally substituted indolyl.
  • the compound has a structural Formula (V):
  • Z 4 is N or CR 5 ;
  • R 5 is halo, -CN, -R,-OR, -S(0) n R, -COOR, -CONR 2 , -NR 2 , optionally substituted aryl, or optionally substituted heteroaryl;
  • R 2 , R 3 and R 4 are each independently selected from H and optionally substituted CI -CI O alkyl;
  • X is O, S, or NR 6 ;
  • Y is O or S or NR 10 ;
  • Z is O or S
  • R 6 and R 10 are each independently selected from H, CN, optionally substituted C 1 -C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C1 -C4 alkoxy, optionally substituted C6-C10 aryl, and optionally substituted heteroaryl;
  • cCy is a carbocyclic ring which is optionally further substituted
  • L 4 is -S(0)-NR-, -S(0) 2 -NR-, -S(0)-0-, -S(0) 2 -0-, -C(0)-NR-, -C(0)-O, -NR-S(O)-, - NR-S(0) 2 -, -O-S(O)-, -0-S(0) 2 -, -NR-C(O)-, or -O-C(O)-;
  • Ar is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, or optionally substituted heterocyclyl;
  • R 7 is H, or optionally substituted C1 -C4 alkyl
  • R 1 is selected from H, optionally substituted C 1 -C10 alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; and
  • each R is independently selected from H and optionally substituted C1 -C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member; and
  • n 0, 1 , or 2;
  • Z 4 is N; R 2 , R 3 , and R 4 are H; Z and Y are O; X is NH; R 1 is cyclopropyl; L 4 is -S(0)-NR-, -S(0) 2 -NR-, -S(0)-0-, -S(0) 2 -0-, -C(0)-NR-, -C(O)- 0-; Ar is optionally substituted aryl or optionally substituted heteroaryl; R 7 is H; and cCy is a cyclohexyl ring.
  • Ar-L 4 - is Ar-S(0)2-NR-.
  • Ar is optionally substituted phenyl.
  • Z 4 is N or CR 5 ;
  • R 5 is halo, -CN, -R, -OR, -S(0) n R, -C(0)OR, -C(0)NR 2 , -NR 2 or -Ar;
  • each R is independently selected from H and optionally substituted C1-C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member, n is 0, 1 , or 2;
  • R 2 , R 3 and R 4 are each independently selected from H and optionally substituted CI -CI O alkyl;
  • X is O, S, or NR 6 ;
  • Y is O, S, or NR 10 ;
  • R 6 and R 10 are each independently selected from H, CN, optionally substituted C 1 -C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C 1 -C4 alkoxy, optionally substituted C6-C 10 aryl, and optionally substituted heteroaryl;
  • Z is O or S
  • Ar is an optionally substituted aryl
  • R 1B is selected from H, optionally substituted C1 -C10 alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, or optionally substituted heteroarylalkyl.
  • each A is independently optionally substituted C1 -C 10 alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted C2-C10 alkenyl, optionally substituted C2-C10 alkynyl, -NR 2 ; -OR, -S(0) n R, -NR-S(0) n R, halo, -CN, -N0 2 , - C(0)OR, -0-C(0)-R, -C(0)NR 2 , -NR-C(0)R, or -C(0)R;
  • c 0, 1 , 2, 3, 4, or 5;
  • each n is independently 0, 1 , or 2;
  • each R is independently H or optionally substituted C1-C4 alkyl; or alternatively, the two
  • R groups taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member.
  • R 3 is -alkylene-0-C(0)-R 3a , wherein R 3a is optionally substituted C1 -C4 alkyl, optionally substituted C1-C4 heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, or optionally substituted heterocyclyl.
  • -C(0)-R 3a is a carboxyl terminus of an amino acid.
  • One embodiment of the amino acid is L- Alanine.
  • the present compound is selected from the species disclosed in the specification.
  • the compounds of Formula (I) or any of its subgenera do not include any of the-compound species, i.e., specific compounds, described in U.S. Patent Application No. 12/946,759.
  • the compounds of Formula (I) do not include any of the compounds listed in Table X below:
  • the invention provides a pharmaceutical composition comprising any of the above-described compounds, admixed with a pharmaceutically acceptable excipient.
  • the invention provides a method to treat cancer, a vascular disorder, inflammation, infection, pain, or an immunological disorder comprising administering to a subject in need of such treatment, an effective amount of any of the above-described compounds.
  • the compounds of the invention are useful as medicaments, and are useful for the manufacture of medicaments, including medicaments to treat conditions disclosed herein, such as cancers, inflammatory conditions, infections, pain, and immunological disorders.
  • treat and “treating” as used herein refer to ameliorating, alleviating, lessening, and removing symptoms of a disease or condition.
  • a candidate molecule or compound described herein may be in a therapeutically effective amount in a formulation or medicament, which is an amount that can lead to a biological effect, such as apoptosis of certain cells (e.g., cancer cells), reduction of proliferation of certain cells, or lead to ameliorating, alleviating, lessening, or removing symptoms of a disease or condition, for example.
  • the terms also can refer to reducing or stopping a cell proliferation rate (e.g., slowing or halting tumor growth) or reducing the number of proliferating cancer cells (e.g. , removing part or all of a tumor).
  • a titre of a microorganism in a system i.e., cell, tissue, or subject
  • a system i.e., cell, tissue, or subject
  • reducing the rate of microbial propagation reducing the number of symptoms or an effect of a symptom associated with the microbial infection, and/or removing detectable amounts of the microbe from the system.
  • microorganisms include but are not limited to virus, bacterium and fungus.
  • the compounds of the invention have activities to modulate protein kinases, in particular CK2 activity and/or Pim activity.
  • the compounds of the invention specifically inhibit the activity of C 2, but not Pim, e.g., more than 100, 90, 80, 70, 60, 50, 40, 30, 20, or 10 fold difference between C 2 inhibition vs. Pim inhibition.
  • the compounds of the invention specifically inhibit the acitivity of Pim, but not CK.2, e.g., more than 100, 90, 80, 70, 60, 50, 40, 30, 20, or 10 fold difference between Pim inhibition vs. C 2 inhibition.
  • the compounds of the invention inhibit the activity of C 2 as well as Pim.
  • the compounds of the invention can be used to modulate the activity of C 2 and/or Pim, e.g. , inhibit the activity of C 2 and/or Pim in a cell, e.g. , in vivo or in vitro.
  • compounds of the invention can be used to modulate the activity of C 2, e.g., inhibit the activity of CK2 without substantially interfering or changing the activity of Pim.
  • compounds of the invention can be used to modulate the activity of Pim, e.g., inhibit the activity of Pim without substantially interfering or changing the activity of C 2.
  • compounds of the invention can be used to modulate the activity of CK2 and Pim, e.g., inhibit the activity of C 2 and Pim.
  • the compounds of the invention are thus useful to treat infections by certain pathogens, including protozoans and viruses.
  • the invention thus provides methods for treating protozoal disorders such as protozoan parasitosis, including infection by parasitic protozoa responsible for neurological disorders such as schizophrenia, paranoia, and encephalitis in immunocompromised patients, as well as Chagas' disease.
  • HIV-1 human immunodeficiency virus type 1
  • HPVs human papilloma viruses
  • HSV herpes simplex virus
  • EBV Epstein-Barr virus
  • human cytomegalovirus hepatitis C and B viruses
  • influenza virus Borna disease virus, adenovirus, coxsackievirus, coronavirus and varicella zoster virus.
  • the methods for treating these disorders comprise administering to a subject in need thereof an effective amount of a compound of the present invention.
  • apoptosis refers to an intrinsic cell self-destruction or suicide program.
  • cells undergo a cascade of events including cell shrinkage, blebbing of cell membranes and chromatic condensation and fragmentation. These events culminate in cell conversion to clusters of membrane-bound particles (apoptotic bodies), which are thereafter engulfed by macrophages.
  • the invention in part provides pharmaceutical compositions comprising at least one compound within the scope of the invention as described herein, and methods of using compounds described herein.
  • the invention in part provides methods for identifying a candidate molecule that interacts with a CK2, which comprises contacting a composition containing a CK.2 protein and a molecule described herein with a candidate molecule and determining whether the amount of the molecule described herein that interacts with the protein is modulated, whereby a candidate molecule that modulates the amount of the molecule described herein that interacts with the protein is identified as a candidate molecule that interacts with the protein.
  • Protein kinases catalyze the transfer of a gamma nhosphate from adenosine triphosphate to a serine or threonine amino acid (serine/threonine protein kinase), tyrosine amino acid (tyrosine protein kinase), tyrosine, serine or threonine (dual specificity protein kinase) or histidine amino acid (histidine protein kinase) in a peptide or protein substrate.
  • methods which comprise contacting a system comprising a protein kinase protein with a compound described herein in an amount effective for modulating (e.g., inhibiting) the activity of the protein kinase.
  • the activity of the protein kinase is the catalytic activity of the protein (e.g., catalyzing the transfer of a gamma phosphate from adenosine triphosphate to a peptide or protein substrate).
  • methods for identifying a candidate molecule that interacts with a protein kinase which comprise:
  • a composition containing a protein kinase and a compound described herein with a candidate molecule under conditions in which the compound and the protein kinase interact, and determining whether the amount of the compound that interacts with the protein kinase is modulated relative to a control interaction between the compound and the protein kinase without the candidate molecule, whereby a candidate molecule that modulates the amount of the compound interacting with the protein kinase relative to the control interaction is identified as a candidate molecule that interacts with the protein kinase.
  • Systems in such embodiments can be a cell-free system or a system comprising cells (e.g., in vitro).
  • the protein kinase, the compound or the molecule in some embodiments is in association with a solid phase.
  • the interaction between the compound and the protein kinase is detected via a detectable label, where in some embodiments the protein kinase comprises a detectable label and in certain embodiments the compound comprises a detectable label.
  • the interaction between the compound and the protein kinase sometimes is detected without a detectable label.
  • compositions of matter comprising a protein kinase and a compound described herein.
  • the protein kinase in the composition is a serine- threonine protein kinase.
  • the protein kinase in the composition is, or contains a subunit (e.g. , catalytic subunit, SH2 domain, SH3 domain) of, C 2.
  • the composition is cell free and sometimes the protein kinase is a recombinant protein.
  • the protein kinase can be from any source, such as cells from a mammal, ape or human, for example.
  • a serine-threonine protein kinase sometimes is a member of a subfamily containing one or more of the following amino acids at positions corresponding to those listed in human CK2: leucine at position 45, methionine at position 163 and isoleucine at position 174.
  • Nucleotide and amino acid sequences for protein kinases and reagents are publicly available (e.g., World Wide Web URLs ncbi.nlm.nih.gov/sites/entrez and Invitrogen.com, each last visited December 2, 2009).
  • the invention also in part provides methods for treating a condition related to aberrant cell proliferation.
  • methods of treating a cell proliferative condition in a subject which comprises administering a compound described herein to a subject in need thereof in an amount effective to treat the cell proliferative condition.
  • the subject may be a research animal (e.g., rodent, dog, cat, monkey), optionally containing a tumor such as a xenograft tumor (e.g., human tumor), for example, or may be a human.
  • a cell proliferative condition sometimes is a tumor, e.g., solid or circulating tumor or non-tumor cancer, including but not limited to, cancers of the colorectum, breast, lung, liver, pancreas, lymph node, colon, prostate, brain, head and neck, skin, liver, kidney, blood, ovary and heart (e.g., leukemia, lymphoma, carcinoma).
  • a tumor e.g., solid or circulating tumor or non-tumor cancer, including but not limited to, cancers of the colorectum, breast, lung, liver, pancreas, lymph node, colon, prostate, brain, head and neck, skin, liver, kidney, blood, ovary and heart (e.g., leukemia, lymphoma, carcinoma).
  • compositions of the invention may be used alone or in combination with anticancer or other agents, such as a palliative agents, that are typically administered to a patient being treated for cancer, as further described herein.
  • methods for treating a condition related to inflammation or pain are provided.
  • methods for treating pain in a subject which comprise administering a compound described herein to a subject in need thereof in an amount effective to treat the pain.
  • methods of treating inflammation in a subject which comprise administering a compound described herein to a subject in need thereof in an amount effective to treat the inflammation.
  • the subject may be a research animal (e.g., rodent, dog, cat, monkey), for example, or may be a human.
  • Conditions associated with inflammation and pain include without limitation acid reflux, heartburn, acne, allergies and allergen sensitivities, Alzheimer's disease, asthma, atherosclerosis, bronchitis, carditis, celiac disease, chronic pain, Crohn's disease, cirrhosis, colitis, dementia, dermatitis, diabetes, dry eyes, edema, emphysema, eczema, fibromyalgia, gastroenteritis, gingivitis, heart disease, hepatitis, high blood pressure, insulin resistance, interstitial cystitis, joint pain/arthritis/rheuma tr > i arthritis, metabolic syndrome (syndrome X), myositis, nephritis, obesity, osteopenia, glomerulonephritis (GN), juvenile cystic kidney disease, and type 1 nephronophthisis (NPHP), osteoporosis, Parkinson's disease, Guam- Parkinson dementia, supranuclear palsy, Ku
  • Methods for determining and monitoring effects of compounds herein on pain or inflammation are known. For example, formalin-stimulated pain behaviors in research animals can be monitored after administration of a compound described herein to assess treatment of pain (e.g. , Li et al., Pain 115(1-2): 182-90 (2005)). Also, modulation of pro-inflammatory molecules ⁇ e.g., IL-8, GRO-alpha, MCP- 1 , TNFalpha and iNOS) can be monitored after administration of a compound described herein to assess treatment of inflammation (e.g., Parhar et al., Int J Colorectal Dis. 22(6): 601 -9 (2006)), for example.
  • methods for determining whether a compound herein reduces inflammation or pain which comprise contacting a system with a compound described herein in an amount effective for modulating (e.g., inhibiting) the activity of a pain signal or inflammation signal.
  • identifying a compound that reduces inflammation or pain comprise: contacting a system with a compound as described herein; and detecting a pain signal or inflammation signal, whereby a compound that modulates the pain signal relative to a control molecule is identified as a compound that reduces inflammation of pain.
  • pain signals are formalin-stimulated pain behaviors and examples of inflammation signals include without limitation a level of a pro-inflammatory molecule.
  • the invention thus in part pertains to methods for modulating angiogenesis in a subject, and methods for treating a condition associated with aberrant angiogenesis in a subject.
  • CK2 has also been shown to play a role in the pathogenesis of atherosclerosis, and may prevent atherogenesis by maintaining laminar shear stress flow.
  • C 2 plays a role in
  • HDACs histone deacetylases
  • the invention provides methods to treat each of these conditions, comprising administering to a subject in need of such treatment an effect amount of a CK2 inhibitor, such as a compound as described herein, such as a compound of Formula (I), (II), (Ila), (lib), (lie), (lid), (III), (Ilia), (Illb), (IV), (V), (VI), or (Via) .
  • a CK2 inhibitor such as a compound as described herein, such as a compound of Formula (I), (II), (Ila), (lib), (lie), (lid), (III), (Ilia), (Illb), (IV), (V), (VI), or (Via) .
  • the invention also in part pertains to methods for modulating an immune response in a subject, and methods for treating a condition associated with an aberrant immune response in a subject.
  • methods for determining whether a compound herein modulates an immune response which comprise contacting a system with a compound described herein in an amount effective for modulating (e.g., inhibiting) an immune response or a signal associated with an immune response.
  • Signals associated with immunomodulatory activity include, e.g., stimulation of T-cell proliferation, suppression or induction of cytokines, including, e.g., interleukins, interferon- ⁇ and TNF.
  • Also provided are methods for treating a condition associated with an aberrant immune response in a subject which comprise administering a compound described herein to a subject in need thereof in an amount effective to treat the condition.
  • Conditions characterized by an aberrant immune response include without limitation, organ transplant rejection, asthma, autoimmune disorders, including rheumatoid arthritis, multiple sclerosis, myasthenia gravis, systemic lupus erythematosus, scleroderma, polymyositis, mixed connective tissue disease (MCTD), Crohn's disease, and ulcerative colitis.
  • an immune response may be modulated by administering a compound herein in combination with a molecule that modulates (e.g., inhibits) the biological activity of an mTOR pathway member or member of a related pathway (e.g., mTOR, PI3 kinase, AKT).
  • a molecule that modulates e.g., inhibits
  • the biological activity of an mTOR pathway member or member of a related pathway e.g., mTOR, PI3 kinase, AKT.
  • the molecule that modulates the biological activity of an mTOR pathway member or member of a related pathway is rapamycin.
  • provided herein is a composition comprising a compound described herein in combination with a molecule that modulates the biological activity of an mTOR pathway member or member of a related pathway, such as rapamycin, for example.
  • the invention provides pharmaceutical compositions (i.e.,
  • compositions can comprise a compound of any of Formula (I), (II), (Ha), (lib), (lie), (lid), (III), (Ilia), (Illb), (IV), (V), (VI), and (Via), as described herein which is admixed with at least one pharmaceutically acceptable excipient or carrier. Frequently, the composition comprises at least two pharmaceutically acceptable excipients or carriers.
  • compositions and methods of the present invention will typically be used in therapy for human patients, they may also be used in veterinary medicine to treat similar or identical diseases.
  • the compositions may, for example, be used to treat mammals, including, but not limited to, primates and domesticated mammals.
  • the compositions may, for example be used to treat herbivores.
  • the compositions of the present invention include geometric and optical isomers of one or more of the drugs, wherein each drug is a racemic mixture of isomers or one or more purified isomers.
  • compositions suitable for use in the present invention include
  • compositions wherein the active ingredients are contained in an effective amount to achieve the intended purpose. Determination of the effective amounts is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein.
  • the compounds of the present invention may exist as pharmaceutically acceptable salts.
  • the present invention includes such salts.
  • pharmaceutically acceptable salts is meant to include salts of active compounds which are prepared with relatively nontoxic acids or bases, depending on the particular substituent moieties found on the compounds described herein.
  • base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Included are base addition salts such as sodium, potassium, calcium, ammonium, organic amino, or magnesium salt, or a similar salt.
  • acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent.
  • acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids, for example, acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p- tolylsulfonic, citric, tartaric, methanesulfonic, and the lik p A ⁇ so included are salts of amino acids such as arginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like (see, for example, Berge et al., "Pharmaceutical Salts", Journal of
  • Examples of applicable salt forms include hydrochlorides, hydrobromides, sulfates, methanesulfonates, nitrates, maleates, acetates, citrates, fumarates, tartrates (eg (+)-tartrates, (-)- tartrates or mixtures thereof, including racemic mixtures), succinates, benzoates and salts with amino acids such as glutamic acid.
  • These salts may be prepared by methods known to those skilled in art.
  • the neutral forms of the compounds are preferably regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner.
  • the parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents.
  • esters in the present invention refer to non-toxic esters, preferably the alkyl esters such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl or pentyl esters, of which the methyl ester is preferred.
  • esters such as phenyl-Ci-5 alkyl may be employed if desired.
  • Ester derivatives of certain compounds may act as prodrugs which, when absorbed into the bloodstream of a warm-blooded animal, may cleave in such a manner as to release the drug form and permit the drug to afford improved therapeutic efficacy.
  • Certain compounds of the present invention can exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to unsolvated forms and are encompassed within the scope of the present invention. Certain compounds of the present invention may exist in multiple crystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated by the present invention and are intended to be within the scope of the present invention.
  • a physiologically acceptable carrier is a formulation to which the compound can be added to dissolve it or otherwise facilitate its administration.
  • physiologically acceptable carriers include, but are not limited to, water, saline, and physiologically buffered saline.
  • the present invention provides compounds that are in a prodrug form.
  • Prodrugs of the compounds described herein are those compounds that readily undergo chemical changes under physiological conditions to provide the compounds of the present invention.
  • prodrugs can be converted to the compounds of the present invention by chemical or biochemical methods in an ex vivo environment. For example, prodrugs can be slowly converted to the compounds of the present invention when placed in a transdermal patch reservoir with a suitable enzyme or chemical reagent.
  • a compound of the present invention can be formulated as a pharmaceutical composition.
  • Such a pharmaceutical composition can then be administered orally, parenterally, by inhalation spray, rectally, or topically in dosage unit formulations containing conventional nontoxic pharmaceutically acceptable carriers, adjuvants, and vehicles as desired.
  • the amount of active ingredient that can be combined with the carrier materials to produce a single dosage form varies depending upon the mammalian host treated and the particular mode of administration.
  • Topical administration can also involve the use of transdermal administration such, as transdermal patches or iontophoresis devices.
  • parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrasternal injection, or infusion techniques. Formulation of drugs is discussed in, for example, Hoover, John E., REMINGTON'S PHARMACEUTICAL
  • sterile injectable preparations for example, sterile injectable aqueous or oleaginous suspensions can be formulated according to the known art using suitable dispersing or wetting agents and suspending agents.
  • the sterile injectable preparation can also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1 ,3-butanediol.
  • acceptable vehicles and solvents that can be employed are water, Ringer's solution, and isotonic sodium chloride solution.
  • sterile, fixed oils are conventionally employed as a solvent or suspending medium.
  • any bland fixed oil can be employed including synthetic mono- or diglycerides.
  • fatty acids such as oleic acid find use in the preparation of injectables.
  • Suppositories for rectal administration of the drug can be prepared by mixing the drug with a suitable nonirritating excipient such as cocoa butter, synthetic mono- di- or triglycerides, fatty acids and polyethylene glycols that are sold at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum and release the drug.
  • Solid dosage forms for oral administration can include capsules, tablets, pills, powders, and granules.
  • the compounds of this invention are ordinarily combined with one or more adjuvants appropriate to the indicated route of administration.
  • a compound of the invention can be admixed with lactose, sucrose, starch powder, cellulose esters of alkanoic acids, cellulose alkyl esters, talc, stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric and sulfuric acids, gelatin, acacia gum, sodium alginate, polyvinylpyrrolidone, and/or polyvinyl alcohol, and then tableted or encapsulated for convenient administration.
  • Such capsules or tablets can contain a controlled- release formulation as can be provided in a dispersion of active compound in
  • the dosage forms can also comprise buffering agents such as sodium citrate, magnesium or calcium carbonate or bicarbonate. Tablets and pills can additionally be prepared with enteric coatings.
  • formulations for parenteral administration can be in the form of aqueous or non-aqueous isotonic sterile injection solutions or suspensions. These solutions and suspensions can be prepared from sterile powders or granules having one or more of the carriers or diluents mentioned for use in the formulations for oral administration.
  • a compound of the invention can be dissolved in water, polyethylene glycol, propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride, and/or various buffers.
  • Other adjuvants and modes of administration are well and widely known in the pharmaceutical art.
  • Liquid dosage forms for oral administration can include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art, such as water.
  • Such compositions can also comprise adjuvants, such as wetting agents, emulsifying and suspending agents, and sweetening, flavoring, and perfuming agents.
  • the dosage regimen utilizing the compounds of the present invention in combination with an anticancer agent is selected in accordance with a variety of factors including type, species, age, weight, sex and medical condition of the pa ⁇ ipnt ; the severity of the condition to be treated; the route of administration; the renal and hepatic function of the patient; and the particular compound or salt or ester thereof employed. A consideration of these factors is well within the purview of the ordinarily skilled clinician for the purpose of determining the therapeutically effective dosage amounts to be given to a person in need of the instant combination therapy.
  • Systemic administration may also include relatively noninvasive methods such as the use of suppositories, transdermal patches, transmucosal delivery and intranasal administration.
  • Oral administration is also suitable for compounds of the invention. Suitable forms include syrups, capsules, tablets, as is understood in the art.
  • a suitable dosage of the compound of the invention for an adult patient will be between 1 and 1000 mg per dose, frequently between 10 and 300 mg, and the dosage may be administered 1 -4 times per day. Dosage levels are dependent on the nature of the condition, drug efficacy, the condition of the patient, the judgment of the practitioner, and the frequency and mode of administration; optimization of such parameters is within the ordinary level of skill in the art.
  • Compounds of the invention may be used alone or in combination with another therapeutic agent.
  • the invention provides methods to treat conditions such as cancer, inflammation and immune disorders by administering to a subject in need of such treatment a therapeutically effective amount of a therapeutic agent useful for treating said disorder and administering to the same subject a therapeutically effective amount of a modulator of the present invention, i.e., a compound of the invention.
  • the therapeutic agent and the modulator may be "co-administered", i.e, administered together, either as separate pharmaceutical compositions or admixed in a single pharmaceutical composition. By “administered together", the therapeutic agent and the modulator may also be administered separately, including at different times and with different frequencies.
  • the modulator may be administered by any known route, such as orally, intravenously, intramuscularly 1 , nasally, and the like; and the therapeutic agent may also be administered by any conventional route. In many embodiments, at least one and optionally both of the modulator and the therapeutic agent may be administered orally.
  • the modulator is an inhibitor, and it may inhibit either one of C 2 and Pirn, or both of them to provide the treatment effects described herein.
  • a "modulator" as described above may be used in combination with a therapeutic agent that can act by binding to regions of DNA that can form certain quadruplex structures.
  • the therapeutic agents have anticancer activity on their own, but their activity is enhanced when they are used in combination with a modulator. This synergistic effect allows the therapeutic agent to be administered in a lower dosage while achieving equivalent or higher levels of at least one desired effect.
  • a modulator may be separately active for treating a cancer.
  • the dosage of a modulator when used in combination with a therapeutic agent, will frequently be two-fold to ten-fold lower than the dosage required when the modulator is used alone to treat the same condition or subject. Determination of a suitable amount of the modulator for use in combination with a therapeutic agent is readily determined by methods known in the art.
  • anticancer agents include, e.g. , classic chemotherapeutic agents, as well as molecular targeted therapeutic agents, biologic therapy agents, and radiotherapeutic agents.
  • the present invention provides, for example, simultaneous, staggered, or alternating treatment.
  • the compound of the invention may be administered at the same time as an anticancer agent, in the same pharmaceutical composition; the compound of the invention may be administered at the same time as the anticancer agent, in separate pharmaceutical compositions; the compound of the invention may be administered before the anticancer agent, or the anticancer agent may be administered before the compound of the invention, for example, with a time difference of seconds, minutes, hours, days, or weeks.
  • a course of therapy with the compound of the invention may be administered, followed by a course of therapy with the anticancer agent, or the reverse order of treatment may be used, and more than one series of treatments with each component may also be used.
  • one component for example, the compound of the invention or the anticancer agent
  • the present compound is administered while the anticancer agent or its derivative products remains in the bloodstream, or the anticancer agent may be administered while the present compound or its derivatives remains in the bloodstream.
  • the second component is administered after all, or most of the first component, or its derivatives, have left the bloodstream of the mammal.
  • the compound of the invention and the anticancer agent may be administered in the same dosage form, e.g., both administered as intravenous solutions, or they may be administered in different dosage forms, e.g. , one compound may be administered topically and the other orally.
  • a person of ordinary skill in the art would be able to discern which combinations of agents would be useful based on the particular characteristics of the drugs and the cancer involved.
  • Anticancer agents useful in combination with the compounds of the present invention may include agents selected from any of the classes known to those of ordinary skill in the art, including, but not limited to, antimicrotubule agents such as diterpenoids and vinca alkaloids; platinum coordination complexes; alkylating agents such as nitrogen mustards,
  • oxazaphosphorines alkylsulfonates, nitrosoureas, and triazenes
  • antibiotic agents such as anthracyclins, actinomycins and bleomycins
  • topoisomerase II inhibitors such as
  • epipodophyllotoxins such as purine and pyrimidine analogues and anti-folate compounds; topoisomerase I inhibitors such as camptothecins; hormones and hormonal analogues; signal transduction pathway inhibitors; nonreceptor tyrosine kinase angiogenesis inhibitors; immunotherapeutic agents; pro-apoptotic agents; and cell cycle signaling inhibitors; and other agents described below.
  • Anti-microtubule or anti-mitotic agents are phase specific agents that are typically active against the microtubules of tumor cells during M or the mitosis phase of the cell cycle.
  • anti-microtubule agents include, but are not limited to, diterpenoids and vinca alkaloids.
  • Plant alkaloid and terpenoid derived agents include mitotic inhibitors such as the vinca alkaloids vinblastine, vincristine, vindesine, and vinorelbine; and microtubule polymer stabilizers such as the taxanes, including, but not limited to paclitaxel, docetaxel, larotaxel, ortataxel, and tesetaxel.
  • Diterpenoids which are derived from natural sources, are phase specific anti - cancer agents that are believed to operate at the G2/ phases of the cell cycle. It is believed that the diterpenoids stabilize the p-tubulin subunit of the microtubules, by binding with this protein. Disassembly of the protein appears then to be inhibited with mitosis being arrested and cell death following.
  • diterpenoids examples include, but are not limited to, taxanes such as paclitaxel, docetaxel, larotaxel, ortataxel, and tesetaxel.
  • Paclitaxel is a natural diterpene product isolated from the Pacific yew tree Taxus brevifolia and is commercially available as an injectable solution TAXOL®.
  • Docetaxel is a semisynthetic derivative of paclitaxel q. v. , prepared using a natural precursor, 10-deacetyl-baccatin III, extracted from the needle of the European Yew tree.
  • Docetaxel is commercially available as an injectable solution as TAXOTERE®.
  • Vinca alkaloids are phase specific anti-neoplastic agents derived from the periwinkle plant. Vinca alkaloids that are believed to act at the M phase (mitosis) of the cell cycle by binding specifically to tubulin. Consequently, the bound tubulin molecule is unable to polymerize into microtubules. Mitosis is believed to be arrested in metaphase with cell death following. Examples of vinca alkaloids include, but are not limited to, vinblastine, vincristine, vindesine, and vinorelbine. Vinblastine, vincaleukoblastine sulfate, is commercially available as VELBAN® as an injectable solution.
  • Vincristine vincaleukoblastine 22-oxo-sulfate
  • ONCOVIN® an injectable solution
  • Vinorelbine is commercially available as an injectable solution of vinorelbine tartrate (NAVELBINE®), and is a
  • Platinum coordination complexes are non-phase specific anti-cancer agents, which are interactive with DNA.
  • the platinum complexes are believed to enter tumor cells, undergo, aquation and form intra- and interstrand crosslinks with DNA causing adverse biological effects to the tumor.
  • Platinum-based coordination complexes include, but are not limited to cisplatin, carboplatin, nedaplatin, oxaliplatin, satraplatin, and (SP-4-3)-(cis)- amminedichloro-[2- methylpyridine] platinum(II).
  • Cisplatin, cis-diamminedic > oplatinum is commercially available as PLATINOL® as an injectable solution.
  • Carboplatin, platinum, diammine [1 , 1 - cyclobutane-dicarboxylate(2-)-0,0'] is commercially available as PARAPLATIN® as an injectable solution.
  • Alkylating agents are generally non-phase specific agents and typically are strong electrophiles. Typically, alkylating agents form covalent linkages, by alkylation, to DNA through nucleophilic moieties of the DNA molecule such as phosphate, amino, sulfhydryl, hydroxyl, carboxyl, and imidazole groups. Such alkylation disrupts nucleic acid function leading to cell death.
  • alkylating agents include, but are not limited to, alkyl sulfonates such as busulfan; ethyleneimine and methylmelamine derivatives such as altretamine and thiotepa; nitrogen mustards such as chlorambucil, cyclophosphamide, estramustine, ifosfamide, mechlorethamine, melphalan, and uramustine; nitrosoureas such as carmustine, lomustine, and streptozocin; triazenes and imidazotetrazines such as dacarbazine, procarbazine, temozolamide, and temozolomide.
  • alkyl sulfonates such as busulfan
  • ethyleneimine and methylmelamine derivatives such as altretamine and thiotepa
  • nitrogen mustards such as chlorambucil, cyclophosphamide, estramustine, ifosfamide, mechlorethamine,
  • Cyclophosphamide 2-[bis(2-chloroethyl)-amino]tetrahydro-2H- 1 ,3,2- oxazaphosphorine 2-oxide monohydrate, is commercially available as an injectable solution or tablets as CYTOXAN®.
  • Melphalan 4-[bis(2-chloroethyl)amino]-L-phenylalanine, is commercially available as an injectable solution or tablets as AL ERAN®.
  • Chlorambucil 4- [bis(2-chloroethyl)amino]-benzenebutanoic acid, is commercially available as LEUKERAN® tablets.
  • Busulfan 1 ,4-butanediol dimethanesulfonate, is commercially available as
  • Carmustine, l ,3-[bis(2-chloroethyl)-l -nitrosourea, is commercially available as single vials of lyophilized material as BiCNU®, 5-(3,3-dimethyl- l -triazeno)- imidazole-4-carboxamide, is commercially available as single vials of material as DTIC-Dome®.
  • alkylating agents include (a) alkylating-like platinum-based chemotherapeutic agents such as cisplatin, carboplatin, nedaplatin, oxaliplatin, satraplatin, and (SP-4-3)-(cis)- amminedichloro-[2-methylpyridine] platinum(II); (b) alkyl sulfonates such as busulfan;
  • ethyleneimine and methylmelamine derivatives such as altretamine and thiotepa
  • nitrogen mustards such as chlorambucil, cyclophosphamide, estramustine, ifosfamide, mechlorethamine, trofosamide, prednimustine, melphalan, and uramustine
  • nitrosoureas such as carmustine, lomustine, fotemustine, nimustine, ranimustine and streptozocin
  • imidazotetrazines such as dacarbazine, procarbazine, temozolamide, and temozolomide.
  • Anti-tumor antibiotics are non-phase specific agents which are believed to bind or intercalate with DNA. This may result in stable DNA com p lexes or strand breakage, which disrupts ordinary function of the nucleic acids, leading to cell death.
  • anti-tumor antibiotic agents include, but are not limited to, anthracyclines such as daunorubicin (including liposomal daunorubicin), doxorubicin (including liposomal doxorubicin), epirubicin, idarubicin, and valrubicin; streptomyces-related agents such as bleomycin, actinomycin, mithramycin, mitomycin, porfiromycin; and mitoxantrone.
  • anthracyclines such as daunorubicin (including liposomal daunorubicin), doxorubicin (including liposomal doxorubicin), epirubicin, idarubicin, and valrubicin
  • Dactinomycin also know as Actinomycin D
  • Daunorubicin (8S-cis-)-8-acetyl- 10-[(3-amino-2,3,6-trideoxy-a-L-lyxohexopyranosy l)oxy]-7,8,9, 10-tetrahydro-6,8, 1 1 - trihydroxy- l-methoxy-5, 12-naphthacenedione hydrochloride, is commercially available as a liposomal injectable form as DAU OXOME® or as an injectable as CERUB1DINE®.
  • Bleomycin a mixture of cytotoxic glycopeptide antibiotics isolated from a strain of Streptomyces verticillus, is commercially available as BLENOXANE®.
  • Topoisomerase inhibitors include topoisomerase I inhibitors such as camptothecin, topotecan, irinotecan, rubitecan, and belotecan; and topoisomerase II inhibitors such as etoposide, teniposide, and amsacrine.
  • Topoisomerase II inhibitors also include, but are not limited to, epipodophyllotoxins, which are phase specific anti-neoplastic agents derived from the mandrake plant.
  • Epipodophyllotoxins typically affect cells in the S and G2 phases of the cell cycle by forming a ternary complex with topoisomerase II and DNA causing DNA strand breaks. The strand breaks accumulate and cell death follows.
  • Examples of epipodophyllotoxins include, but are not limited to, etoposide, teniposide, and amsacrine.
  • Etoposide, 4'-demethyl-epipodophyllotoxin 9[4,6-0- (R)-ethylidene- -D-. glucopyranoside] is commercially available as an injectable solution or capsules as VePESID® and is commonly known as VP- 16.
  • Teniposide, 4'-demethyl- epipodophyllotoxin 9[4,6-0-(R )-thenylidene-P-D-glucopyranoside], is commercially available as an injectable solution as VUMON® and is commonly known as VM-26.
  • topoisomerase I inhibitors include, but are not limited to camptothecin and camptothecin derivatives, topotecan, irinotecan, rubitecan, belotecan and the various optical forms (i.e., (R), (S) or (R,S)) of 7-(4-methylpiperazino-methylene)- 10, 1 1 - ethylenedioxy-camptothecin, as described in U.S. Patent 6,063,923; 6, 100,273; 5,342,947; 5,559,235; and 5,491 ,237.
  • Irinotecan is a derivative of camptothecin which binds, along with its active metabolite 8N-38, to the topoisomerase I-DNA complex.
  • Topotecan HC1 (S)-10-[(dimethylamino)methyl]-4-ethyl- 4,9-dihydroxy-l H-pyrano[3',4',6,7]indolizino[l ,2-b]quinoIine-3, 14-(4H, 12H)-dione monohydrochloride, is commercially available as the injectable solution HYCA TIN®.
  • Anti-metabolites include (a) purine analogs such as fludarabine, cladribine,
  • pyrimidine analogs such as fluorouracil, gemcitabine, capecitabine, cytarabine, azacitidine, edatrexate, floxuridine, and troxacitabine;
  • antifolates such as methotrexate, pemetrexed, raltitrexed, and trimetrexate.
  • Anti-metabolites also include thymidylate synthase inhibitors, such as fluorouracil, raltitrexed, capecitabine, floxuridine and pemetrexed; and ribonucleotide reductase inhibitors such as claribine, clofarabine and fludarabine.
  • Antimetabolite neoplastic agents are phase specific anti-neoplastic agents that typically act at S phase (DNA synthesis) of the cell cycle by inhibiting DNA synthesis or by inhibiting purine or pyrimidine base synthesis and thereby limiting DNA synthesis. Consequently, S phase does not proceed and cell death follows.
  • Antimetabolites include purine analogs, such as fludarabine, cladribine, chlorodeoxyadenosine, clofarabine, mercaptopurine, pentostatin, erythrohydroxynonyladenine, fludarabine phosphate and thioguanine; pyrimidine analogs such as fluorouracil, gemcitabine, capecitabine, cytarabine, azacitidine, edatrexate, floxuridine, and troxacitabine; antifolates, such as methotrexate, pemetrexed, raltitrexed, and trimetrexate.
  • purine analogs such as fludarabine, cladribine, chlorodeoxyadenosine, clofarabine, mercaptopurine, pentostatin, erythrohydroxynonyladenine, fludarabine phosphate and thioguanine
  • pyrimidine analogs such as fluorouracil, gem
  • Cytarabine 4-amino- l -p-D-arabinofuranosyl-2 ( 1 H)- pyrimidinone, is commercially available as CYTOSAR-U® and is commonly known as Ara-C.
  • Mercaptopurine 1 ,7-dihydro-6H-purine-6-thione monohydrate, is commercially available as PURINETHOL®.
  • Thioguanine, 2-amino-l , 7-dihydro-6H-purine-6-thione is commercially available as TABLOID®.
  • Gemcitabine 2'-deoxy-2', 2'-difluorocytidine monohydrochloride (p- isomer), is commercially available as GE ZAR®.
  • Hormonal therapies include (a) androgens such as fluoxymesterone and testolactone; (b) antiandrogens such as bicalutamide, cyproterone, flutamide, and nilutamide; (c) aromatase inhibitors such as aminoglutethimide, anastrozole, exemestane, formestane, and letrozole;
  • corticosteroids such as dexamethasone and prednison e p
  • estrogens such as diethylstilbestrol
  • antiestrogens such as fulvestrant, raloxifene, tamoxifen, and toremifine;
  • LHRH agonists and antagonists such as buserelin, goserelin, leuprolide, and triptorelin;
  • progestins such as medroxyprogesterone acetate and megestrol acetate
  • thyroid hormones such as levothyroxine and liothyronine.
  • Hormones and hormonal analogues are useful compounds for treating cancers in which there is a relationship between the hormone(s) and growth and/or lack of growth of the cancer.
  • hormones and hormonal analogues useful in cancer treatment include, but are not limited to, androgens such as fluoxymesterone and testolactone; antiandrogens such as bicalutamide, cyproterone, flutamide, and nilutamide;
  • aromatase inhibitors such as aminoglutethimide, anastrozole, exemestane, formestane, vorazole, and letrozole; corticosteroids such as dexamethasone, prednisone and prednisolone; estrogens such as diethylstilbestrol; antiestrogens such as fulvestrant, raloxifene, tamoxifen, toremifine, droloxifene, and iodoxyfene, as well as selective estrogen receptor modulators (SERMS) such those described in U.S. Patent Nos.
  • SERMS selective estrogen receptor modulators
  • 5a-reductases such as finasteride and dutasteride
  • progestins such as medroxyprogesterone acetate and megestrol acetate
  • thyroid hormones such as levothyroxine and liothyronine.
  • Signal transduction pathway inhibitors are those inhibitors, which block or inhibit a chemical process which evokes an intracellular change, such as cell proliferation or
  • Signal trahduction inhibitors useful in the present invention include, e.g., inhibitors of receptor tyrosine kinases, non-receptor tyrosine kinases, SH2/SH3 domain blockers, serine/threonine kinases, phosphotidyl inositol-3 kinases, myo-inositol signaling, and Ras oncogenes.
  • Molecular targeted agents include (a) receptor tyrosine kinase ('RTK') inhibitors, such as inhibitors of EGFR, including erlotinib, gefitinib, and neratinib; inhibitors of VEGFR including vandetanib, semaxinib, and cediranib; and inhibitors of PDGFR; further included are RTK inhibitors that act at multiple receptor sites such as lapatinib, which inhibits both EGFR and HER2, as well as those inhibitors that act at each of C-kit, PDGFR and VEGFR, including but not limited to axitinib, sunitinib, sorafenib and toceranib; also included are inhibitors of BCR- ABL, c-kit and PDGFR, such as imatinib; (b) FKBP bindin g agents, such as an immunosuppressive macrolide antibiotic, including bafilomycin, rapamycin (sirolimus) and everolimus;
  • adapalene bexarotene, trans-retinoic acid, 9-cis-retinoic acid, and N-(4-hydroxyphenyl)retinamide
  • phenotype-directed therapy agents including monoclonal antibodies such as alemtuzumab, bevacizumab, cetuximab, ibritumomab tiuxetan, rituximab, and trastuzumab
  • immunotoxins such as gemtuzumab ozogamicin
  • radioimmunoconjugates such as 1311-tositumomab
  • cancer vaccines such as 1311-tositumomab
  • Protein tyrosine kinases catalyse the phosphorylation of specific tyrosyl residues in various proteins involved in the regulation of cell growth.
  • Such protein tyrosine kinases can be broadly classified as receptor or non-receptor kinases.
  • Receptor tyrosine kinases are transmembrane proteins having an extracellular ligand binding domain, a transmembrane domain, and a tyrosine kinase domain.
  • Receptor. tyrosine kinases are involved in the regulation of cell growth and are sometimes termed growth factor receptors.
  • Growth factor receptors include, for example, epidermal growth factor receptor (EGFr), platelet derived growth factor receptor (PDGFr), erbB2, erbB4, vascular endothelial growth factor receptor (VEGFr), tyrosine kinase with immunoglobulin-like and epidermal growth factor homology domains (TIE-2), insulin growth factor -I (IGFI) receptor, macrophage colony stimulating factor (cfms), BTK, ckit, cmet, fibroblast growth factor (FGF) receptors, Trk receptors (TrkA, TrkB, and TrkC), ephrin (eph) receptors, and the RET protooncogene.
  • EGFr epidermal growth factor receptor
  • PDGFr platelet derived growth factor receptor
  • erbB2 erbB4
  • VEGFr vascular endothelial growth factor receptor
  • TIE-2 vascular endothelial growth factor receptor
  • TIE-2 t
  • inhibitors of growth receptors include ligand antagonists, antibodies, tyrosine kinase inhibitors and anti-sense oligonucleotides.
  • Growth factor receptors and agents that inhibit growth factor receptor function are described, for instance, in Kath, John C, Exp. Opin. Ther. Patents (2000) 10(6):803-818; Shawver et al., Drug Discov. Today ( 1997), 2(2):50-63; and Lofts, F. J. et a!., "Growth factor receptors as targets", New Molecular Targets for Cancer Chemotherapy, ed. Workman, Paul and Kerr, David, CRC press 1994, London.
  • receptor tyrosine kinase inhibitors include, but are not limited to, sunitinib, erlotinib, gefitinib, and imatinib.
  • Tyrosine kinases which are not growth factor receptor kinases are termed non-receptor tyrosine kinases.
  • Non-receptor tyrosine kinases useful in the present invention, which are targets or potential targets of anti-cancer drugs include cSrc, Lck, Fyn, Yes, Jak, cAbl, FAK (Focal adhesion kinase), Brutons tyrosine kinase, and Bcr-Abl.
  • non-receptor kinases and agents which inhibit non-receptor tyrosine kinase function are described in Sinh, S. and Corey, S.J., J. Hematotherapy & Stem Cell Res. (1999) 8(5): 465 - 80; and Bolen, J.B., Brugge, J.S., Annual Review of Immunology. (1997) 15: 371-404.
  • SH2/SH3 domain blockers are agents that disrupt SH2 or SH3 domain binding in a variety of enzymes or adaptor proteins including, PI3-K p85 subunit, Src family kinases, adaptor molecules (She, Crk, Nek, Grb2) and Ras-GAP.
  • SH2/SH3 domains as targets for anti-cancer drugs are discussed in Smithgall, T.E., J Pharmacol. Toxicol. Methods. (1995), 34(3): 125-32.
  • Inhibitors of Serine/Threonine Kinases including MAP kinase cascade blockers which include blockers of Raf kinases (rafk), Mitogen or Extracellular Regulated Kinase (MEKs), and
  • Extracellular Regulated Kinases ERKs
  • Protein kinase C family member blockers including blockers of PKCs (alpha, beta, gamma, epsilon, mu, lambda, iota, zeta).
  • IkB kinase family IKKa, IKKb
  • PKB family kinases PKB family kinases
  • AKT kinase family members zeta
  • TGF beta receptor kinases TGF beta receptor kinases.
  • Serine/Threonine kinases and inhibitors thereof are described in Yamamoto, T., Taya, S., Kaibuchi, K., J. Biochemistry.
  • Myo-inositol signaling inhibitors such as phospholipase C blockers and Myoinositol analogues.
  • signal inhibitors are described in Powis, G., and Kozikowski A, (1994) New Molecular Targets for Cancer
  • Ras Oncogene inhibitors include inhibitors of farnesyltransferase, geranyl-geranyl transferase, and CAAX proteases as well as anti-sense oligonucleotides, ribozymes and immunotherapy. Such inhibitors have been shown to block ras activation in cells containing wild type mutant ras, thereby acting as antiproliferation agents. Ras oncogene inhibition is discussed in Scharovsky, O.G., Rozados, V.R, Gervasoni, SI, Matar, P., J. Biomed. Sci.
  • antibody antagonists to receptor kinase ligand binding may also serve as signal transduction inhibitors.
  • This group of signal transduction pathway inhibitors includes the use of humanized antibodies to the extracellular ligand binding domain of receptor tyrosine kinases.
  • Imclone C225 EGFR specific antibody ⁇ see Green, M.C. et al., Cancer Treat. Rev., (2000) 26(4): 269-286
  • Herceptin® erbB2 antibody ⁇ see Stern, DF, Breast Cancer Res. (2000) 2(3): 176-183
  • 2CB VEGFR2 specific antibody see Brekken, R.A. et al., Cancer Res. (2000) 60(18):51 17-24).
  • Non-receptor kinase angiogenesis inhibitors may also find use in the present invention.
  • Inhibitors of angiogenesis related VEGFR and TIE2 are discussed above in regard to signal transduction inhibitors (both receptors are receptor tyrosine kinases).
  • Angiogenesis in general is linked to erbB2/EGFR signaling since inhibitors of erbB2 and EGFR have been shown to inhibit angiogenesis, primarily VEGF expression.
  • the combination of an erbB2/EGFR inhibitor with an inhibitor of angiogenesis makes sense.
  • non-receptor tyrosine kinase inhibitors may be used in combination with the EGFR/erbB2 inhibitors of the present invention.
  • anti-VEGF antibodies which do not recognize VEGFR (the receptor tyrosine kinase), but bind to the ligand; small molecule inhibitors of integrin (alphav beta3) that will inhibit angiogenesis; endostatin and angiostatin (non-RT ) may also prove useful in combination with the disclosed erb family inhibitors.
  • VEGFR the receptor tyrosine kinase
  • small molecule inhibitors of integrin alphav beta3
  • endostatin and angiostatin non-RT
  • erb family inhibitors See Bruns, CJ et al., Cancer Res. (2000), 60(1 1 ): 2926-2935; Schreiber AB, Winkler ME, & Derynck R., Science (1986) 232(4755): 1250- 53; Yen L. et al., Oncogene (2000) 19(31 ): 3460-9).
  • Agents used in immunotherapeutic regimens may also be useful in combination with the compounds of formula (I).
  • immunologic strategies to generate an immune response against erbB2 or EGFR. These strategies are generally in the realm of tumor vaccinations.
  • the efficacy of immunologic approaches r" av be greatly enhanced through combined inhibition of erbB2/EGFR signaling pathways using a small molecule inhibitor. Discussion of the immunologic/tumor vaccine approach against erbB2/EGFR are found in Reilly RT, et a!., Cancer Res. (2000) 60( 13):3569-76; and Chen Y, et al., Cancer Res. (1998)
  • Agents used in pro-apoptotic regimens may also be used in the combination of the present invention.
  • Members of the Bcl-2 family of proteins block apoptosis. Upregulation of bcl-2 has therefore been linked to chemoresistance.
  • Cell cycle signalling inhibitors inhibit molecules involved in the control of the cell cycle.
  • a family of protein kinases called cyclin dependent kinases (CDKs) and their interaction with a family of proteins termed cyclins controls progression through the eukaryotic cell cycle. The coordinate activation and inactivation of different cyclin/CDK complexes is necessary for normal progression through the cell cycle.
  • CDKs cyclin dependent kinases
  • Several inhibitors of cell cycle signalling are under development. For instance, examples of cyclin dependent kinases, including CDK2, CDK4, and CDK6 and inhibitors for the same are described in, for instance, RosaniaGR & Chang Y-T., Exp. Opin. Ther. Patents (2000) 10(2):215-30.
  • FKBP binding agents such as the FKBP binding agents
  • immunosuppressive macrolide antibiotic rapamycin
  • gene therapy agents antisense therapy agents, and gene expression modulators
  • retinoids and rexinoids e.g. , adapalene, bexarotene, trans-retinoic acid, 9-cisretinoic acid, and N-(4 hydroxyphenyl)retinamide
  • retinoids and rexinoids e.g. , adapalene, bexarotene, trans-retinoic acid, 9-cisretinoic acid, and N-(4 hydroxyphenyl)retinamide
  • phenotype-directed therapy agents including: monoclonal antibodies such as alemtuzumab, bevacizumab, cetuximab, ibritumomab tiuxetan, rituximab, and trastuzumab; immunotoxins such as gemtuzumab ozogamicin, radioimmunoconjugates such as 131 -tositumomab; and cancer vaccines.
  • Anti-tumor antibiotics include (a) anthracyclines such as daunorubicin (including liposomal daunorubicin), doxorubicin (including liposorr* 1 d oxorubicin), epirubicin, idarubicin, and valrubicin; (b) streptomyces-related agents such as bleomycin, actinomycin, mithramycin, mitomycin, porfiromycin; and (c) anthracenediones, such as mitoxantrone and pixantrone.
  • anthracyclines such as daunorubicin (including liposomal daunorubicin), doxorubicin (including liposorr* 1 d oxorubicin), epirubicin, idarubicin, and valrubicin
  • streptomyces-related agents such as bleomycin, actinomycin, mithramycin, mitomycin, porfiro
  • Anthracyclines have three mechanisms of action: intercalating between base pairs of the DNA/RNA strand; inhibiting topoiosomerase II enzyme; and creating iron-mediated free oxygen radicals that damage the DNA and cell membranes.
  • Anthracyclines are generally characterized as topoisomerase II inhibitors.
  • Monoclonal antibodies include, but are not limited to, murine, chimeric, or partial or fully humanized monoclonal antibodies.
  • Such therapeutic antibodies include, but are not limited to antibodies directed to tumor or cancer antigens either on the cell surface or inside the cell.
  • Such therapeutic antibodies also include, but are not limited to antibodies directed to targets or pathways directly or indirectly associated with C 2.
  • Therapeutic antibodies may further include, but are not limited to antibodies directed to targets or pathways that directly interact with targets or pathways associated with the compounds of the present invention.
  • therapeutic antibodies include; but are not limited to anticancer agents such as Abagovomab, Adecatumumab, Afutuzumab, Alacizumab pegol, Alemtuzumab, Altumomab pentetate, Anatumomab mafenatox, Apolizumab, Bavituximab, Belimumab, Bevacizumab, Bivatuzumab mertansine, Blinatumomab, Brentuximab vedotin, Cantuzumab mertansine, Catumaxomab, Cetuximab, Citatuzumab communicatingox, Cixutumumab, Clivatuzumab tetraxetan, Conatumumab, Dacetuzumab, Detumomab, Ecromeximab, Edrecolomab, Elotuzumab, Epratuzumab
  • Lucatumumab Lumiliximab, Mapatumumab, Matuzumab, Milatuzumab, Mitumomab, Nacolomab tafenatox, Naptumomab estafenatox, Necitumumab, Nimotuzumab, Ofatumumab, Olaratumab, Oportuzumab monatox, Oregovomab, Panitumumab, Pemtumomab, Pertuzumab, Pintumomab, Pritumumab, Ramucirumab, Rilotumumab, Rituximab, Robatumumab,
  • such therapeutic antibodies include, alemtuzumab, bevacizumab, cetuximab, daclizumab, gemtuzumab, ibritumomab tiuxetan, pantitu m, ,r nab, rituximab, tositumomab, and trastuzumab; in other embodiments, such monoclonal antibodies include alemtuzumab, bevacizumab, cetuximab, ibritumomab tiuxetan, rituximab, and trastuzumab; alternately, such antibodies include daclizumab, gemtuzumab, and pantitumumab.
  • therapeutic antibodies useful in the treatment of infections include but are not limited to Afelimomab, Efungumab, Exbivirumab, Felvizumab, Foravirumab, Ibalizumab, Libivirumab, Motavizumab, Nebacumab, Pagibaximab, Palivizumab, Panobacumab, Rafivirumab,
  • therapeutic antibodies can be useful in the treatment of inflammation and/or autoimmune disorders, including, but are not limited to, Adalimumab, Atlizumab,
  • Atorolimumab Aselizumab, Bapineuzumab, Basiliximab, Benralizumab, Bertilimumab,
  • Otelixizumab Pascolizumab, Priliximab, Reslizumab, Rituximab, Rontalizumab, Rovelizumab, Ruplizumab, Sifalimumab, Siplizumab, Solanezumab, Stamulumab, Talizumab, Tanezumab, Teplizumab, Tocilizumab, Toralizumab, Ustekinumab, Vedolizumab, Vepalimomab,
  • such therapeutic antibodies include, but are not limited to adalimumab, basiliximab, certolizumab pegol, eculizumab, efalizumab, infliximab, muromonab-CD3, natalizumab, and omalizumab.
  • the therapeutic antibody can include abciximab or ranibizumab.
  • a therapeutic antibody is non-conjugated, or is conjugated with a radionuclide, cytokine, toxin, drug-activating enzyme or a drug-filled liposome.
  • Akt inhibitors include l L6-Hydroxymethyl-chiro-inositol-2-(R)-2-0-methyl-3-0- octadecyl-stt-glycerocarbonate, SH-5 (Calbiochem Cat. No. 124008), SH-6 (Calbiochem Cat. No. Cat. No. 124009), Calbiochem Cat. No. 12401 1 , Triciribine (NSC 154020, Calbiochem Cat. No.
  • PX-866 (acetic acid (1 S,4E, 10R, 1 l R, 13S, 14R)-[4- diallylaminomethylene-6-hydroxy-l -methoxymethyl-10, 13-dimethy 1-3,7, 17-trioxo-
  • inhibitors such as, for example, BEZ-235, PX-866, D 106669, CAL- 101 , GDC0941 , SF 1 126, SF2523 are also identified in the art as PI3 /mTOR inhibitors; additional examples, such as PI- 103 [3-[4-(4- morpholinylpyrido[3',2':4,5]furo[3,2-d]pyrimidin-2-yl]phenol hydrochloride] are well-known to those of skill in the art. Additional well-known PI3K inhibitors include LY294002 [2-(4- morpholinyl)-8-phenyl-4H-l -benzopyran-4-one] and wortmannin.
  • mTOR inhibitors known to those of skill in the art include temsirolimus, deforolimus, sirolimus, everolimus, zotarolimus, and biolimus A9.
  • a representative subset of such inhibitors includes temsirolimus, deforolimus, zotarolimus, and biolimus A9.
  • HDAC inhibitors include (i) hydroxamic acids such as Trichostatin A, vorinostat (suberoylanilide hydroxamic acid (SAHA)), panobinostat (LBH589) and belinostat (PXD101 ) (ii) cyclic peptides, such as trapoxin B, and depsipeptides, such as romidepsin (NSC 630176),
  • benzamides such as MS-275 (3-pyridylmethyl-N- ⁇ 4-[(2-aminophenyl)-carbamoyl]-benzyl ⁇ - carbamate), CI994 (4-acetylamino-N-(2aminophenyl)-benzamide) and MGCD0103 (N-(2- aminophenyl)-4-((4-(pyridin-3-yl)pyrimidin-2-ylamino)methyl)benzamide), (iv) electrophilic ketones, (v) the aliphatic acid compounds such as phenylbutyrate and valproic acid.
  • Hsp90 inhibitors include benzoquinone ansamycins such as geldanamycin, 17-DMAG
  • Miscellaneous agents include altretamine, arsenic trioxide, gallium nitrate, hydroxyurea, levamisole, mitotane, octreotide, procarbazine, suramin, thalidomide, lenalidomide, photodynamic compounds such as methoxsalen and sodium porfimer, and proteasome inhibitors such as bortezomib.
  • Biologic therapy agents include: interferons such as interferon-a2a and interferon-a2b, and interleukins such as aldesleukin, denileukin diftitox, and oprelvekin.
  • combination therapies including the use of protective or adjunctive agents, including: cytoprotective agents such as armifostine, dexrazonxane, and mesna, phosphonates such as peridronate and zoledronic acid, and stimulating factors such as epoetin, darbepoetin, filgrastim, PEG-filgrastim, and sargramostim, are also envisioned.
  • cytoprotective agents such as armifostine, dexrazonxane, and mesna
  • phosphonates such as mbodronate and zoledronic acid
  • stimulating factors such as epoetin, darbepoetin, filgrastim, PEG-filgrastim, and sargramostim
  • the compounds of the present invention can be synthesized using methods, techniques, and materials known to those of skill in the art, such as described, for example, in March, ADVANCED ORGANIC CHEMISTRY 4 th ed., (Wiley 1992); Carey and Sundberg,
  • Preparation of the present compounds may include one or more steps of protection and deprotection (e.g. , the formation and removal of acetal groups).
  • Guidance for selecting suitable protecting groups can be found, for example, in Greene & Wuts, "Protective Groups in Organic Synthesis," Wiley Interscience, 1999.
  • the preparation may include various purifications, such as column chromatography, flash chromatography, thin-layer chromatography (TLC), recrystallization, distillation, high-pressure liquid chromatography (HPLC) and the like.
  • Table 1 A The molecules described in the following table (Table 1 A) were prepared using the procedures similar to those exemplified in Example 1. All compounds were characterized by LCMS.
  • Table I B shows the biological activities of the compounds listed in Table 1A as well as Examples 1 , 2, 4, 6, 7, 10, and 12. That is, Compounds 1A to IO in Table I B are the compounds listed in Table 1 A and Examples 1 , 2, 4, 6, 7, 10, and 12.
  • Hydantoin (20 mg, 0.2 mmol) and 3-chloro-4-(7-(cyclopropylamino)-3-formyl-6- methylpyrazolo[l ,5-a]pyrimidin-5-ylamino)benzonitrile (30 mg, 0.1 mmol) were dissolved in ethanol (0.5 mL) along with piperidine (20 uL, 0.2 mmol). The reaction was heated at 85 °G in the microwave for 2 hours. The reaction was cooled to r.t., diluted with water, and the precipitate was collected and washed with water, (1 : 1 ) ethanol/water, then ethanol.
  • reaction mixture was heated in microwave for 10 minutes at 1 10 °C, diluted with 1 : 1 mixture of MeOH and dichloromethane and purified using preparative TLC to provide (Z)-5-((5-((2'-chloro-4-fluorobiphenyl-2-yl)methylamino)-7- (cyclopropylamino)pyrazolo[l ,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione as solid.
  • LCMS (ES) > 95% pure, m/z 518 [M+H]+.
  • Table 2A The compounds described in the following Table 2A were prepared using procedures similar to those exemplified in Example 17. All compounds were characterized by LCMS.
  • Table 2B shows the biological activities of the compounds listed in Table 2A and Examples 14 to 18. That is, Compounds I P to 1 Z are compounds listed in Table 2A and Examples 14 to 18. Table 2A.
  • Table 3 A The compounds described in the following Table 3 A were prepared using procedures similar to those exemplified in the above examples. All compounds were characterized by LCMS.
  • Table 3B shows the biological activities of the compounds listed in Table 3A and Examples 20 to 23. That is, Compounds I P to 1Z are compounds listed in Table 3A and Examples 20 to 23.
  • Table 4A The compounds described in the following Table 4A were prepared using procedures similar to those exemplified in Examples 26 and 27. All compounds were characterized by LCMS. Table 4B shows the biological activities of the compounds listed in Table 4A as well as Examples 25 to 27.
  • Table 5A The compounds described in the following Table 5A were prepared using procedures similar to those exemplified in Example 31 . All compounds were characterized by LCMS. Table 5D shows the biological activities of the compounds listed in Table 5A, 5B, and 5C as well as Examples 29 to 31 , 32, and 33.
  • Example 32 Similar to those exemplified in Example 32. All compounds were characterized by LCMS. Table 5D shows the biological activities of the compounds listed in Table 5B.
  • 3-Methylimidazolidine-2,4-dione was prepared according to the literature procedure setforth in Eur. JOC 2002, 1763.
  • 5-Chloro-6-(7-(cyclopropylamino)-3-forrnylpyrazolo[l ,5- o]pyrimidin-5-ylamino)nicotinonitrile 2,2,2-trifluoroacetate 38 mg, 0.1 mmol
  • EtOH mL
  • 3-methylimidazolidine-2,4-dione 17 mg, 0.15 mmol
  • piperidine 25 iL, 0.25 mmol
  • Table 6 shows the biological activities of certain selected compounds of Exampl and 42 to 48.
  • Table 7 shows the biological activity of Example 51 . That is, Compound 3U in Table 7 is Example 51 .
  • Table 7A The compounds in the following Table 7A were prepared using procedures described in Example 52 by using the appropriate starting boronic acids. Compounds were prepared by HPLC purification and concentrated down in Genevac. Some required further TLC purification (2% MeOH in DCM).
  • Table 7B shows the biological activities of the compounds listed in Table 7A. That is, Compounds 3V to 41 in Table 7B are the compounds listed in Table 7A.
  • Example 54 by using the appropriate starting boronic acids. Compounds were prepared by HPLC purification and concentrated down in Genevac. Some required further TLC purification (2% MeOH in DCM). Table 8B shows the biological activities of the compounds listed in Table 8A and Example 54.
  • Table 9A The compounds in the following Table 9A were prepared using procedures described in Example 56 by using the appropriate starting boronic acids. Compounds were prepared by HPLC purification and concentrated down in Genevac. Some required further TLC purification (2% MeOH in DCM). Table 9B shows the biological activities of the compounds listed in Table 9A.
  • Genevac evaporation provided (Z)-5-((4- (cyclopropylamino)-2-((5-ethylpyridin-2-yl)methoxy)pyrazolo[l ,5-a][l ,3,5]triazin-8- yl)methylene)imidazolidine-2,4-dione as a solid (12 mg).
  • reaction mixture was stirred at rt for 1 hour, filtered through a PTFE filter, and purified by mass- directed LC/MS to provide (Z)-5-((4-(cyclopropylamino)pyrazolo[l ,5-a][l ,3,5]triazin-8- yl)methylene)imidazolidine-2,4-dione as the TFA salt.
  • Table 1 1 A The compounds in the following Table 1 1 A were prepared using procedures described in Example 59 by using the appropriate starting boronic acids. Compounds were prepared by HPLC purification and concentrated down in Genevac. Table 1 IB shows the biological activities of the compounds listed in Table 1 1 A as well as Examples 59 to 62. That is, Compounds 5R to 5X in Table 1 I B are the compounds listed in Table 1 1 A and Examples 59 to 62. Table 1 1 A.
  • Example 60 Synthesis of (ZVmethyl 2-(4-r4-(cvclopropylaminoV8-((2,5-dioxoimidazolidin-4- ylidene ' )methyl ' )pyrazolo 1.5-airi ,3,5]triazin-2-ylamino ' )phenoxy)acetate
  • Example 63 Synthesis of N-fnr ⁇ - ⁇ -rcvclopro ylamino S-frZ t ' .S-dioxoimidazolidin ⁇ - ylidene ' )methyl ' )pyrazolori ,5-a ri .3.51triazin-2-ylamino ' )cvclohexyl ' )-4- fluorobenzenesulfonamide
  • Example 64 Synthesis of 4-cvano-N-(4- ⁇ 4-cyclopropylamino-8- 2,5-dioxo-imidazolidin-(4Z)- ylidenemethyll-pyrazolo K5-al l ,3,5]triazin-2-ylamino)-cyclohexyO-benzenesulfonamide
  • N, N-Dimethyl-5-(5-methylthiophen-2-yl)pyrazolo[l ,5-a]pyrimidin-7-amine (66 mg, 0.26 mmol) was dissolved in anhydrous DMF (3.3 mL) and the solution was cooled to 0 °C.
  • POC (30 ⁇ , 0.33 mmol) was added dropwise at a rate such that the internal temperature was maintained ⁇ 5 °C.
  • the solution immediately turned yellow and after the addition a precipitate formed.
  • the reaction was gradually warmed to 21 °C by removal of the ice bath.
  • the solution became homogeneous after ⁇ 4 h.
  • the solution was added to ice water (30 mL), and 6 ⁇ NaOH was added to pH 1 1.
  • PdC dppf-ChhC 43 mg, 0.05 mmol were added sequentially. The reaction was placed in an 85 °C oil bath. After 6 h, the solution was diluted with brine (50 mL) and extracted with EtOAc (3 x 50 mL). The organics were dried over MgSC>4, filtered, and concentrated in vacuo.
  • Trifluoroacetic acid (1 mL) was added and the solution changed color from yellow to orange. After 1 h, the solution was concentrated under a stream of air. Trituration with Et20 (5 mL) gave 7-(methylamino)-5-(5-methylthiophen-2-yl)pyrazolo[l ,5-a]pyrimidine-3-carbaldehyde (44 mg, 88%) as an off white solid.
  • the reaction was gradually warmed to 21 °C by removal of the ice bath. After 22 h, the solution was added to ice water (20 mL), and 6N NaOH was added to pH 11. After 1 h of stirring the solution was filtered. The filter cake was purified via flash column chromatography (10-15% EtOAc/hexanes) to yield /er/-butyl 3-formyl-5-(5-methylthiophen-2-yl)pyrazolo[l,5- o]pyrimidin-7-yl(isopropyl)carbamate (50 mg, 51%) as a yellow solid.
  • Trifluoroacetic acid (0.6 mL) was added and the solution changed color from bright yellow to dark orange/red. After 1 h, the solution was concentrated under a stream of air to an orange oil. Trituration with Et20 (3 mL) gave 7-(isopropylamino)-5-(5-methylthiophen-2-yl)pyrazolo[l ,5- a]pyrimidine-3-carbaldehyde (28 mg, 77%) as a tan solid.
  • reaction mixture was stirred under microwave heating at 1 10°C for 2 hours, filtered through a PTFE filter , and purified by mass-directed LC/MS to provide (Z)-5-(f7-f4-fluorophenylamino)-5-(5- methylthiophen-2-yl)pyrazolo[l ,5-a] as the
  • DME and 2M a2CC>3 were degassed with a stream of nitrogen for 10 minutes prior to use.
  • 7e -butyl 5-chloro-3-formylpyrazolo[l ,5-cr]pyrimidin-7-yl(cyclopropyl)carbamate (672 mg, 2 mmol) and thiophene-2-boronic acid (384 mg, 3 mmol) were dissolved in DME (12 mL) under an atmosphere of nitrogen.
  • Pd(PPli 3 )4 (231 mg, 0.2 mmol

Abstract

The invention provides compounds that inhibit protein kinase CK2 activity (CK2 activity), and compositions containing such compounds. These compounds and compositions are useful for treating proliferative disorders such as cancer, as well as other kinase-associated conditions including inflammation, pain, and certain immunological disorders, and have the following general formula:

Description

PYRAZOLOPYRIMIDINES AND RELATED HETEROCYCLES AS CK2 INHIBITORS
CROSS-REFERENCE TO RELATED APPLICATIONS
This application claims priority to U.S. Provisional Patent Application No. 61 /494,660, filed June 8, 201 1 , which is incorporated by reference in its entirety. This application is related to U.S. Patent Application No. 12/946,759, filed November 15, 2010 and entitled
"PYRAZOLOPYRIMIDINES AND RELATED HETEROCYCLES AS KINASE
INHIBITORS", the contents of which are hereby incorporated by references in their entireties for all purposes.
FIELD OF THE INVENTION
The invention relates in part to molecules having certain biological activities that include, but are not limited to, inhibiting cell proliferation, and modulating certain protein kinase activities. Molecules of the invention modulate, e.g. , Protein Kinase CK2 (called CK2 herein) and are useful to treat conditions associated directly or indirectly with CK2 activities, e.g. , cancers, inflammatory conditions, infectious disorders, pain, immunological disorders, a neurodegenerative disorder (such as Alzheimer's disease and Parkinson's disease), etc. The invention also relates in part to methods for using such compounds, and pharmaceutical compositions containing these compounds.
BACKGROUND
Protein kinase CK2 (formerly called Casein kinase II, referred to herein as "CK2") is a ubiquitous and highly conserved protein serine/threonine kinase. The holoenzyme is typically found in tetrameric complexes consisting of two catalytic (alpha and/or alpha') subunits and two regulatory (beta) subunits. CK2 has a number of physiological targets and participates in a complex series of cellular functions including the maintenance of cell viability. The level of CK2 in normal cells is tightly regulated, and it has long been considered to play a role in cell growth and proliferation. Inhibitors of CK2 that are useful for treating certain types of cancers are described in PCT/US2007/077464, PCT/US2008/074820, PCT/US2009/35609.
The prevalence and importance of CK2, as well as an evolutionary analysis of its sequence, suggest it is an ancient enzyme on the evolutionary scale; its longevity may explain why it has become important in so many biochemical processes, and why CK2 from hosts have even been co-opted by infectious pathogens (e.g., viruses, protozoa) as an integral part of their survival and life cycle biochemical systems. These same characteristics explain why inhibitors of CK2 are believed to be useful in a variety of medical treatments as discussed herein. Because CK2 is central to many biological processes, as summarized by Guerra & Issinger, Curr. Med. Chem. , 2008, 15: 1870-1886, inhibitors of CK2, including the compounds described herein, should be useful in the treatment of a variety of diseases and disorders.
Cancerous cells show an elevation of CK2, and recent evidence suggests that CK2 exerts potent suppression of apoptosis in cells by protecting regulatory proteins from caspase-mediated degradation. The anti-apoptotic function of CK2 may contribute to its ability to participate in transformation and tumorigenesis. In particular, CK2 has been shown to be associated with acute and chronic myelogenous leukemia, lymphoma and multiple myeloma. In addition, enhanced CK2 activity has been observed in solid tumors of the colon, rectum and breast, squamous cell carcinomas of the lung and of the head and neck (SCCHN), adenocarcinomas of the lung, colon, rectum, kidney, breast, and prostate. Inhibition of C 2 by a small molecule is reported to induce apoptosis of pancreatic cancer cells, and hepatocellular carcinoma cells (HegG2, Hep3, HeLa cancer cell lines); and CK2 inhibitors dramatically sensitized RMS (Rhabdomyosarcoma) tumors toward apoptosis induced by TRAIL. Thus an inhibitor of CK2 alone, or in combination with TRAIL or a ligand for the TRAIL receptor, would be useful to treat RMS, the most common soft-tissue sarcoma in children. In addition, elevated C 2 has been found to be highly correlated with aggressiveness of neoplasias, and treatment with a CK2 inhibitor of the invention should thus reduce tendency of benign lesions to advance into malignant ones, or for malignant ones to metastasize.
Unlike other kinases and signaling pathways, where mutations are often associated with structural changes that cause loss of regulatory control, increased C 2 activity level appears to be generally caused by upregulation or overexpression of the active protein rather than by changes that affect activation levels. Guerra and Issinger postulate this may be due to regulation by aggregation, since activity levels do not correlate well with mRNA levels. Excessive activity of CK2 has been shown in many cancers, including SCCHN tumors, lung tumors, breast tumors, and others. Id. Elevated CK2 activity in colorectal carcinomas was shown to correlate with increased malignancy. Aberrant expression and activity of CK2 have been reported to promote increase nuclear levels of NF-kappaB in breast cancer cells. C 2 activity is markedly increased in patients with AML and CML during blast crisis, indicating that an inhibitor of C 2 should be particularly effective in these conditions. Multiple myeloma cell survival has been shown to rely on high activity of CK2, and inhibitors of CK2 were cytotoxic to MM cells.
The literature provides clear evidence that inhibition of CK2 correlates with efficacy against tumor cells. For example, a CK2 inhibitor inhibited growth of murine pi 90 lymphoma cells. Its interaction with Bcr/Abl has been reported to play an important role in proliferation of Bcr/Abl expressing cells, indicating inhibitors of C 2 may be useful in treatment of Bcr/Abl- positive leukemias. Inhibitors of C 2 have been shown to inhibit progression of skin papillomas, prostate and breast cancer xenografts in mice, and to prolong survival of transgenic mice that express prostate-promoters. Id.
The role of C 2 in various non-cancer disease processes has been recently reviewed. See Guerra & Issinger, Curr. Med. Chem. , 2008, 15: 1870-1886. Increasing evidence indicates that CK2 is involved in critical diseases of the central nervous system, including, for example, Alzheimer's disease, Parkinson's disease, and rare neurodegenerative disorders such as Guam- Parkinson dementia, chromosome 18 deletion syndrome, progressive supranuclear palsy, Kuf s disease, or Pick's disease. It is suggested that selective CK2-mediated phosphorylation of tau proteins may be involved in progressive neurodegeneration of Alzheimer's disease. In addition, recent studies suggest that CK2 plays a role in memory impairment and brain ischemia, the latter effect apparently being mediated by C 2's regulatory effect on the PI3K survival pathways.
C 2 has also been shown to be involved in the modulation of inflammatory disorders, for example, acute or chronic inflammatory pain, glomerulonephritis, and autoimmune diseases, including, e.g., multiple sclerosis (MS), systemic lupus erythematosus, rheumatoid arthritis, and juvenile arthritis. It positively regulates the function of the serotonin 5-HT3 receptor channel, activates heme oxygenase type 2, and enhances the activity of neuronal nitric oxide synthase. A selective CK2 inhibitor was reported to strongly reduce pain response of mice when
administered to spinal cord tissue prior to pain testing. It phosphorylates secretory type IIA phospholipase A2 from synovial fluid of RA patients, and modulates secretion of DE (a nuclear DNA-binding protein), which is a proinflammato™ molecule found in synovial fluid of patients with juvenile arthritis. Thus, inhibition of CK2 is expected to control progression of inflammatory pathologies such as those described here, and the inhibitors disclosed herein have been shown to effectively treat pain in animal models.
Protein kinase C 2 has also been shown to play a role in disorders of the vascular system, such as, e.g., atherosclerosis, laminar shear stress, and hypoxia. CK2 has also been shown to play a role in disorders of skeletal muscle and bone tissue, such as cardiomyocyte hypertrophy, impaired insulin signaling and bone tissue mineralization. In one study, inhibitors of CK2 were effective at slowing angiogenesis induced by growth factor in cultured cells.
Moreover, in a retinopathy model, a CK2 inhibitor combined with octreotide (a somatostatin analog) reduced neovascular tufts; thus, the CK2 inhibitors described herein would be effective in combination with a somatostatin analog to treat retinopathy.
CK2 has also been shown to phosphorylate GSK, troponin and myosin light chain; thus, C 2 is important in skeletal muscle and bone tissue physiology, and is linked to diseases affecting muscle tissue.
Evidence suggests that CK2 is also involved in the development and life cycle regulation of protozoal parasites, such as, for example, Theileria parva, Trypanosoma cruzi, Leishmania donovani, Herpetomonas muscarum muscarum, Plasmodium falciparum, Trypanosoma brucei, Toxoplasma gondii and Schistosoma mansoni. Numerous studies have confirmed the role of CK.2 in regulation of cellular motility of protozoan parasites, essential to invasion of host cells. Activation of CK2 or excessive activity of CK2 has been shown to occur in hosts infected with Leishmania donovani, Herpetomonas muscarum muscarum, Plasmodium falciparum,
Trypanosoma brucei, Toxoplasma gondii and Schistosoma mansoni. Indeed, inhibition of CK2 has been shown to block infection by T. cruzi.
C 2 has also been shown to interact with and/or phosphorylate viral proteins associated with human immunodeficiency virus type 1 (HIV-1 ), human papilloma virus, and herpes simplex virus, in addition to other virus types (e.g. , human cytomegalovirus, hepatitis C and B viruses, Borna disease virus, adenovirus, coxsackievirus, coronavirus, influenza, and varicella zoster virus). C 2 phosphorylates and activates HIV- 1 reverse transcriptase and proteases in vitro and in vivo, and promotes pathogenicity of simian-human immunodeficiency virus (SHIV), a model for HIV. Inhibitors of CK2 are thus able to reduce pathogenic effects of a model of HIV infection. C 2 also phosphorylates numerous proteins in herpes simplex virus and numerous other viruses, and some evidence suggests viruses have adopted CK2 as a phosphorylating enzyme for their essential life cycle proteins. Inhibition of CK2 is thus expected to deter infection and progression of viral infections, which rely upon the host's CK2 for their own life cycles.
CK2 is unusual in the diversity of biological processes that it affects, and it differs from most kinases in other ways as well: it is constitutively active, it can use ATP or GTP, and it is elevated in most tumors and rapidly proliferating tissues. In addition, while many kinase inhibitors affect multiple kinases, increasing the likelihood of off-target effects or variability between individual subjects, CK2's unique structural features enable discovery of highly CK2- specific inhibitors. For all of these reasons, CK2 is a particularly interesting target for drug development, and the invention provides highly effective inhibitors of C 2 that are useful in treating a variety of different diseases and disorders mediated by or associated with excessive, aberrant or undesired levels of CK2 activity.
The current invention provides novel compounds of Formula (I), (II), (Ila), (lib), (lie), (lid), (III), (Ilia), (Illb), (IV), (V), (VI), and (Via) including any compound species thereof, as well as the salt, solvate, and/or prodrug thereof, and pharmaceutical compositions containing these compounds or their salt, solvate, and/or prodrugs thereof. The compounds of the present invention show surprisingly greater activity on CK2 and reduced Pirn activity, and thus are advantageously used to treat conditions sensitive to C 2 inhibition such as those described herein. The present compounds are therefore useful to treat conditions mediated by or associated with excessive activity of CK2, with reduced likelihood of off-target effects caused by inhibition of other kinases.
DISCLOSURE OF THE INVENTION
The present invention in part provides chemical compounds having certain biological activities that include, but are not limited to, inhibiting cell proliferation, inhibiting angiogenesis, and modulating protein kinase activities. These molecules, such as compounds of Formula (I), (II), (Ila), (lib), (lie), (lid), (III), (Ilia), (Illb), (IV), (V), (VI), or (Via) including any compound species thereof, modulate protein kinase CK2 (CK2) and/or PIM activity, and are typically more selective for CK2 activity over other kinases. These compounds affect biological functions that include but are not limited to, inhibiting gamma phosphate ransfer from ATP to a protein or peptide substrate, inhibiting angiogenesis, inhibiting cell proliferation and inducing cell apoptosis, for example. The present invention also in part provides methods for preparing chemical compounds, and analogs thereof, and methods of using these compounds. Also provided are compositions comprising these molecules in combination with other materials, including other therapeutic agents, and methods for using such compositions.
Also provided herein are pharmaceutical compositions comprising a compound of the present invention as described herein and at least one pharmaceutically acceptable carrier or excipient, or two or more pharmaceutically acceptable carriers and/or excipients.
Pharmaceutical compositions comprising at least one of these compounds can be utilized in methods of treatment such as those described herein.
The compounds of the present invention as described herein bind to and inhibit certain kinase proteins, which is believed to be the basis for their pharmaceutical activity. In certain embodiments, the protein is a C 2 protein, such as a CK2 protein comprising the amino acid sequence of SEQ ID NO: l , 2 or 3 or a substantially identical variant thereof, for example.
SEQ ID NO: l fNP 001886; casein kinase II alpha 1 subunit isoform a iHomo sapiens!)
msgpvpsrar vytdvnthrp reywdyeshv vewgnqddyq lvrklgrgky sevfeainit nnekvvvkil kpvkkkkikr eikilenlrg gpniitladi vkdpvsrtpa lvfehvnntd 121 fkqlyqtltd ydirfymyei lkaldychsm gimhrdvkph nvmidhehrk lrlidwglae 181 fyhpgqeynv rvasryfkgp ellvdyqmyd ysldmwslgc mlasmifrke pffhghdnyd 241 qlvriakvlg tedlydyidk ynieldprfn dilgrhsrkr werfvhsenq hlvspealdf 301 ldkllrydhq srltareame hpyfytvvkd qarmgsssmp ggstpvssan mmsgissvpt 361 psplgplags pviaaanplg mpvpaaagaq q SEQ ID NO:2 ΓΝΡ 808227; casein kinase II alpha 1 subunit isoform a fHomo sapiens!)
msgpvpsrar vytdvnthrp reywdyeshv vewgnqddyq lvrklgrgky sevfeainit nnekvvvkil kpvkkkkikr eikilenlrg gpniitladi vkdpvsrtpa lvfehvnntd 121 fkqlyqtltd ydirfymyei lkaldychsm gimhrdvkph nvmidhehrk lrlidwglae 181 fyhpgqeynv rvasryfkgp ellvdyqmyd ysldmwslgc mlasmifrke pffhghdnyd 241 qlvriakvlg tedlydyidk ynieldprfn dilgrhsrkr werfvhsenq hlvspealdf 301 ldkllrydhq srltareame hpyfytvvkd qarmgsssmp ggstpvssan mmsgissvpt 361 psplgplags pviaaanplg mpvpaaagaq q SEQ ID NO:3 ΝΡ 808228; casein kinase II alpha 1 subunit isoform b THomo sapiensT)
myeilkaldy chsmgimhrd vkphnvmidh ehrklrlidw glaefyhpgq eynvrvasry fkgpellvdy qmydysldmw slgcmlasmi frkepffhgh dnydqlvria kvlgtedlyd 121 yidkynield prfndilgrh srkrwerfvh senqhlvspe aldfldkllr ydhqsrltar 181 eamehpyfyt vvkdqarmgs ssmpggstpv ssanmmsgis svptpsplgp lagspviaaa 241 nplgmpvpaa agaqq
Substantially identical variants of these include proteins having at least 90% sequence homology with one of these, in one embodiment at least 90% sequence identity; and having at least 50% of the level of in vitro kinase activity of the specified sequence under typical assay conditions.
The invention includes methods to modulate the activity of CK2 protein, either in vitro or ex vivo. Suitable methods comprise contacting a system comprising the protein with a compound described herein in an amount effective for modulating the activity of the protein. In certain embodiments the activity of the protein is inhibited, and sometimes the protein is a CK2 protein comprising the amino acid sequence of SEQ ID NO: l , SEQ ID NO:2 or SEQ ID NO:3 or a substantially identical variant thereof, for example. In certain embodiments the C 2 is in a cell or tissue; in other embodiments, it can be in a cell-free system.
Provided also are methods for inhibiting cell proliferation, which comprise contacting cells with a compound described herein in an amount effective to inhibit proliferation of the cells. The cells sometimes are in a cell line, such as a cancer cell line (e.g., breast cancer, prostate cancer, pancreatic cancer, lung cancer, hemopoietic cancer, colorectal cancer, skin cancer, ovary cancer cell line), for example. In some embodiments, the cancer cell line is a breast cancer, prostate cancer or pancreatic cancer cell line. The cells sometimes are in a tissue, can be in a subject, at times are in a tumor, and sometimes are in a tumor in a subject. In certain embodiments, the method further comprises inducing cell apoptosis. Cells sometimes are from a subject having macular degeneration.
Also provided are methods for treating a condition related to aberrant cell proliferation, which comprise administering a compound described herein to a subject in need thereof in an amount effective to treat the cell proliferative condition. In certain embodiments the cell proliferative condition is a tumor-associated cancer, e.g., a solid or circulating tumor. The cancer sometimes is cancer of the breast, prostate, pancreas, lung, colorectum, skin, or ovary. In some embodiments, the cell proliferative condition is a non-tumor cancer, such as a
hematopoietic cancer, for example, including leukemias, e.g., multiple myeloma and lymphomas. The cell proliferative condition is macular degeneration in some embodiments.
The invention also includes methods for treating cancer or an inflammatory disorder or other disorders described herein that are mediated by excessive activity of one or more of these kinases, in a subject in need of such treatment, comprising: administering to the subject a therapeutically effective amount of a therapeutic agent useful for treating such disorder; and administering to the subject a molecule described herein, e.g., a compound inhibits CK2 in an amount that is effective to enhance a desired effect of the therapeutic agent. In certain embodiments, the molecule that inhibits CK2 is a compound of Formula (I), (II), (Ila), (lib), (lie), (lid), (III), (Ilia), (Illb), (IV), (V), (VI), (Via), or any compound species thereof, or a pharmaceutically acceptable salt, solvate, and/or prodrug thereof. In certain embodiments, the desired effect of the therapeutic agent that is enhanced by the molecule that inhibits CK2 is an increase in apoptosis in at least one type of cell. In certain embodiments, the cell is a cancer cell and the compound is a compound of Formula (I), (II), (Ila), (lib), (lie), (lid), (III), (Ilia), (Illb), (IV), (V), (VI), (Via), or any compound species thereof, that is a potent inhibitor (IC-50 less than about 100 nM, for example) of C 2. In one embodiment, the compound has an IC-50 on Pim of less than about 30 nM, and is selective for CK2 over Pim kinases. In certain embodiments, the IC-50 for inhibition of C 2 is lower by at least a factor of ten than activity on Pim; in preferred embodiments, the compound has an IC-50 for CK2 that is lower than its IC-50 for at least one of Pim-1 , Pim-2 and Pim-3 by about 100-fold or more.
In some embodiments, the therapeutic agent and the molecule that inhibits C 2 are administered at substantially the same time. The therapeutic agent and molecule that inhibits C 2 sometimes are used concurrently by the subject. The therapeutic agent and the molecule that inhibits C 2 can be combined into one pharmaceutical composition in certain embodiments; in other embodiments they are admistered as separate compositions.
Also provided are compositions of matter comprising a compound described herein and an isolated protein. The protein sometimes is a C 2 protein, such as a C 2 protein comprising the amino acid sequence of SEQ ID NO: l, SEQ ID NO:? nr SEQ ID NO:3 or a substantially identical variant thereof, for example. In some embodiments, the protein is a Pirn protein.
Certain compositions comprise a compound described herein in combination with a cell. The cell may be from a cell line, such as a cancer cell line. In the latter embodiments, the cancer cell line is sometimes a breast cancer, prostate cancer, pancreatic cancer, lung cancer, hematopoietic cancer, colorectal cancer, skin cancer, or ovary cancer cell line.
These and other embodiments of the invention are described in the description that follows.
MODES OF CARRYING OUT THE INVENTION
Compounds of the present invention exert biological activities that include, but are not limited to, inhibiting cell proliferation, reducing angiogenesis, preventing or reducing inflammatory responses and pain, and modulating certain immune responses. Such compounds modulate CK2 activity, as demonstrated by the data herein. Such compounds therefore can be utilized in multiple applications by a person of ordinary skill in the art. For example, compounds described herein can be used, for example, for (i) modulation of protein kinase activity (e.g., CK2 activity), (ii) modulation of cell proliferation, (Hi) modulation of apoptosis, and/or (iv) treatments of cell proliferation related disorders (e.g. , administration alone or co-administration with another molecule). In particular, the compounds of Formula (I), (II), (Ha), (lib), (lie), (lid), (III), (Ilia), (Illb), (IV), (V), (VI), or (Via) including any compound species thereof, can be used to modulate CK2 activity, in vitro or in vivo, and to treat disorders associated with excessive or undesirable levels of CK2 activity, including cancers, certain inflammatory disorders, vascular disorders, certain skeletal and muscle disorders, and infections such as protozoal parasite infections and some viral infections.
Definitions:
The terms "a" and "an" do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced item. The terms "a" and "an" are used interchangeably with "one or more" or "at least one". The term "or" or "and/or" is used as a function word to indicate that two words or expressions are to be taken together or individually. The terms "comprising", "having", "including", and "containing" are to be construed as open-ended terms (i.e., meaning "including, but not limited to"). The endpoints of all ranges directed to the same component or property are inclusive and independently combinable.
The terms "compound(s) of the invention", "these compounds", "such compound(s)", "the compound(s)", and "the present compound(s)" refer to compounds encompassed by structural formulae disclosed herein, e.g., Formula (I), (II), (Ila), (lib), (lie), (lid), (III), (Ilia), (Illb), (IV), (V), (VI), and (Via), and include any specific compounds within these formulae whose structure is disclosed herein. Compounds may be identified either by their chemical structure and/or chemical name. When the chemical structure and chemical name conflict, the chemical structure is determinative of the identity of the compound. Furthermore, the present compounds can modulate, i.e., inhibit or enhance, the biological activity of a C 2 protein, a Pim protein or both, and thereby is also referred to herein as a "modulator(s)" or "CK2 and/or Pim modulator(s)". Compounds of Formula (I), (II), (Ila), (lib), (lie), (lid), (III), (Ilia), (Illb), (IV), (V), (VI), and (Via), including any specific compounds, i.e., species, described herein are exemplary "modulators".
The compounds described herein may contain one or more chiral centers and/or double bonds and therefore, may exist as stereoisomers, such as double-bond isomers (i. e. , geometric isomers such as E and Z), enantiomers or diastereomers. The invention includes each of the isolated stereoisomeric forms (such as the enantiomerically pure isomers, the E and Z isomers, etc.) as well as mixtures of stereoisomers in varying degrees of chiral purity or percetange of E and Z, including racemic mixtures, mixtures of diastereomers, and mixtures of E and Z isomers. Accordingly, the chemical structures depicted herein encompass all possible enantiomers and stereoisomers of the illustrated compounds including the stereoisomerically pure form (e.g. , geometrically pure, enantiomerically pure or diastereomerically pure) and enantiomeric and stereoisomeric mixtures. Enantiomeric and stereoisomeric mixtures can be resolved into their component enantiomers or stereoisomers using separation techniques or chiral synthesis techniques well known to the skilled artisan. The invention includes each of the isolated stereoisomeric forms as well as mixtures of stereoisomers in varying degrees of chiral purity, including racemic mixtures. It also encompasses the various diastereomers. Other structures may appear to depict a specific isomer, but that is merely for convenience, and is not intended to limit the invention to the depicted isomer unless otherwise noted. When the chemical name does not specify the isomeric form of the compound, it denotes any one of the possible isomeric forms or a mixtures of those isomeric forms of the compound.
The compounds may also exist in several tautomeric forms, and the depiction herein of one tautomer is for convenience only, and is also understood to encompass other tautomers of the form shown. Accordingly, the chemical structures depicted herein encompass all possible tautomeric forms of the illustrated compounds. The term "tautomer" as used herein refers to isomers that change into one another with great ease so that they can exist together in equilibrium: For example, ketone and enol are two tautomeric forms of one compound. In another example, a substituted 1 ,2,4-triazole derivative may exist in at least three tautomeric forms as shown below:
RT1 is H or optionally substituted alkyl, RT2 is an optionally substituted aryl.
Figure imgf000012_0001
The descriptions of compounds of the present invention are limited by principles of chemical bonding known to those skilled in the art. Accordingly, where a group may be substituted by one or more of a number of substituents, such substitutions are selected so as to comply with principles of chemical bonding and to give compounds which are not inherently unstable and/or would be known to one of ordinary skill in the art as likely to be unstable under ambient conditions, such as aqueous, neutral, and several known physiological conditions. For example, a heterocycloalkyl or heteroaryl is attached to the remainder of the molecule via a ring heteroatom in compliance with principles of chemical bonding known to those skilled in the art thereby avoiding inherently unstable compounds.
Unless otherwise stated, structures depicted herein are also meant to include compounds which differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures except for the replacement of a hydrogen by a deuterium or tritium, or the replacement of a carbon by 13C- or 1 C-enriched carbon are within the scope of this invention. The compounds of the present invention may also contain unnatural proportions of atomic isotopes at one or more of atoms that constitute such compounds. For example, the compounds may be radiolabeled with radioactive isotopes, such as for example tritium (3H), iodine-125 (l25I) or carbon- 14 (14C). All isotopic variations of the compounds of the present invention, whether radioactive or not, are encompassed within the scope of the present invention.
The compounds of the invention often have ionizable groups so as to be capable of preparation as salts. In that case, wherever reference is made to the compound, it is understood in the art that a pharmaceutically acceptable salt may also be used. These salts may be acid addition salts involving inorganic or organic acids or the salts may, in the case of acidic forms of the compounds of the invention be prepared from inorganic or organic bases. Frequently, the compounds are prepared or used as pharmaceutically acceptable salts prepared as addition products of pharmaceutically acceptable acids or bases. Suitable pharmaceutically acceptable acids and bases are well-known in the art, such as hydrochloric, sulphuric, hydrobromic, acetic, lactic, citric, or tartaric acids for forming acid addition salts, and potassium hydroxide, sodium hydroxide, ammonium hydroxide, caffeine, various amines, and the like for forming basic salts. Methods for preparation of the appropriate salts are well-established in the art. In some cases, the compounds may contain both an acidic and a basic functional group, in which case they may have two ionized groups and yet have no net charge. Standard methods for the preparation of pharmaceutically acceptable salts and their formulations are well known in the art, and are disclosed in various references, including for example, "Remington: The Science and Practice of Pharmacy", A. Gennaro, ed., 20th edition, Lippincott, Williams & Wilkins, Philadelphia, PA.
"Solvate", as used herein, means a compound formed by solvation (the combination of solvent molecules with molecules or ions of the solute), or an aggregate that consists of a solute ion or molecule, i.e., a compound of the invention, with one or more solvent molecules. When water is the solvent, the corresponding solvate is "hydrate". Examples of hydrates include, but are not limited to, hemihydrate, monohydrate, dihydrate, trihydrate, hexahydrate, etc. It should be understood by one of ordinary skill in the art that the pharmaceutically acceptable salt, and/or prodrug of the present compound may also exist in a solvate form. The solvate is typically formed via hydration which is either part of the preparation of the present compound or through natural absorption of moisture by the anhydrous compound of the present invention.
The term "ester" means any ester of a present compound in which any of the -COOH functions of the molecule is replaced by a -COOR function, in which the R moiety of the ester is any carbon-containing group which forms a stable ester moiety, including but not limited to alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, »<-vlalkyl, heterocyclyl, heterocyclylalkyl and substituted derivatives thereof. The hydrolysable esters of the present compounds are the compounds whose carboxyls are present in the form of hydrolysable ester groups. That is, these esters are pharmaceutically acceptable and can be hydrolyzed to the corresponding carboxyl acid in vivo. These esters may be conventional ones, including lower alkanoyloxyalkyl esters, e.g., pivaloyloxymethyl and 1-pivaloyloxyethyl esters; lower alkoxycarbonylalkyl esters, e.g., methoxycarbonyloxymethyl, 1 -ethoxycarbonyloxyethyl, and 1- isopropylcarbonyloxyethyl esters; lower alkoxymethyl esters, e.g., methoxymethyl esters, lactonyl esters, benzofuran keto esters, thiobenzofuran keto esters; lower alkanoylaminomethyl esters, e.g., acetylaminomethyl esters. Other esters can also be used, such as benzyl esters and cyano methyl esters. Other examples of these esters include: (2,2-dimethyl-l- oxypropyloxy)methyl esters; (lRS)-l-acetoxy ethyl esters, 2-[(2-methylpropyloxy)carbonyl]-2- pentenyl esters, l-[[(l-methylethoxy)carbonyl]- oxy]ethyl esters; isopropyloxycarbonyloxyethyl esters, (5-methy l-2-oxo-l,3- dioxole-4-yl) methyl esters, l-[[(cyclohexyloxy)carbonyl]oxy]ethyl esters; 3,3-dimethyl-2-oxobutyI esters. It is obvious to those skilled in the art that hydrolysable esters of the compounds of the present invention can be formed at free carboxyls of said compounds by using conventional methods. Representative esters include pivaloyloxymethyl esters, isopropyloxycarbonyloxyethyl esters and (5-methyl-2-oxo-l ,3-dioxole-4-yl)methyl esters.
The term "prodrug" refers to a precursor of a pharmaceutically active compound wherein the precursor itself may or may not be pharmaceutically active but, upon administration, will be converted, either metabolically or otherwise, into the pharmaceutically active compound or drug of interest. For example, a prodrug can be an ester, ether, or amide form of a pharmaceutically active compound. Various types of prodrugs have been prepared and disclosed for a variety of pharmaceuticals. See, for example, Bundgaard, H. and Moss, J., J. Pharm. Sci. 78: 122-126 (1989). Thus, one of ordinary skill in the art knows how to prepare these prodrugs with commonly employed techniques of organic synthesis.
"Protecting group" refers to a grouping of atoms that when attached to a reactive functional group in a molecule masks, reduces or prevents reactivity of the functional group. Examples of protecting groups can be found in Greene & Wuts, "Protective Groups in Organic Synthesis," Wiley Interscience, 1999 and Harrison et al., "Compendium of Synthetic Organic Methods", Vols. 1 -8 (John Wiley and Sons, 1971-1996). Representative amino protecting groups include, but are not limited to, formyl, acetyl, trifl"r>roacetyl, benzyl, benzyloxycarbonyl ("CBZ"), tert-butoxycarbonyl ("Boc"), trimethylsilyl ("TMS"), 2-trimethylsilyl-ethanesulfonyl ("SES"), trityl and substituted trityl groups, allyloxycarbonyl, 9-fluorenylmethyloxycarbonyl ("FMOC"), nitro-veratryloxycarbonyl ("NVOC") and the like. Representative hydroxy protecting groups include, but are not limited to, those where the hydroxy group is either acylated or alkylated such as benzyl, and trityl ethers as well as alkyl ethers, tetrahydropyranyl ethers, trialkylsilyl ethers and allyl ethers.
As used herein, "pharmaceutically acceptable" means suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended use within the scope of sound medical judgment.
"Excipient" refers to a diluent, adjuvant, vehicle, or carrier with which a compound is administered.
An "effective amount" or "therapeutically effective amount" is the quantity of the present compound in which a beneficial outcome is achieved when the compound is administered to a patient or alternatively, the quantity of compound that possesses a desired activity in vivo or in vitro. In the case of proliferative disorders, a beneficial clinical outcome includes reduction in the extent or severity of the symptoms associated with the disease or disorder and/or an increase in the longevity and/or quality of life of the patient compared with the absence of the treatment. For example, for a subject with cancer, a "beneficial clinical outcome" includes a reduction in tumor mass, a reduction in the rate of tumor growth, a reduction in metastasis, a reduction in the severity of the symptoms associated with the cancer and/or an increase in the longevity of the subject compared with the absence of the treatment. The precise amount of compound administered to a subject will depend on the type and severity of the disease or condition and on the characteristics of the patient, such as general health, age, sex, body weight and tolerance to drugs. It will also depend on the degree, severity and type of proliferative disorder. The skilled artisan will be able to determine appropriate dosages depending on these and other factors.
As used herein, the terms "alkyl," "alkenyl" and "alkynyl" include straight-chain, branched-chain and cyclic monovalent hydrocarbyl radicals, and combinations of these, which contain only C and H when they are unsubstituted. Examples include methyl, ethyl, isobutyl, cyclohexyl, cyclopentylethyl, 2-propenyl, 3-butynyl, and the like. The total number of carbon atoms in each such group is sometimes described herein, e. v., when the group can contain up to ten carbon atoms it can be represented as.l-lOC or as C1-C10 or Cl-10. When heteroatoms (N, O and S typically) are allowed to replace carbon atoms as in heteroalkyl groups, for example, the numbers describing the group, though still written as e.g., C1-C6, represent the sum of the number of carbon atoms in the group plus the number of such heteroatoms that are included as replacements for carbon atoms in the backbone of the ring or chain being described. Where a ring is included, it is understood that the group contains at least three carbon atoms as a 3- membered ring is the smallest size for a ring.
Typically, the alkyl, alkenyl and alkynyl substituents of the invention contain 1 -lOC (alkyl) or 2-10C (alkenyl or alkynyl), or 3- IOC when a ring is included. In one embodiment, they contain 1-8C (alkyl) or 2-8C (alkenyl or alkynyl) or 3-8C when a ring is included.
Sometimes they contain 1-4C (alkyl) or 2-4C (alkenyl or alkynyl). A single group can include more than one type of multiple bond, or more than one multiple bond; such groups are included within the definition of the term "alkenyl" when they contain at least one carbon-carbon double bond, and are included within the term "alkynyl" when they contain at least one carbon-carbon triple bond; provided, however, that the presence of multiple bonds does not produce an aromatic ring.
Alkyl, alkenyl and alkynyl groups are often optionally substituted to the extent that such substitution makes sense chemically.
"Optionally substituted" as used herein indicates that the particular group or groups being described may have no non-hydrogen substituents, or the group or groups may have one or more non-hydrogen substituents. If not otherwise specified, the total number of such substituents that may be present is equal to the number of H atoms present on the unsubstituted form of the group being described. Where an optional substituent is attached via a double bond, such as a carbonyl oxygen (=0), the group takes up two available alences, so the total number of substituents that may be included is reduced according to the number of available valences.
"Substituted," when used to modify a specified group or radical, means that one or more hydrogen atoms of the specified group or radical are each, independently of one another, replaced with the same or different substituent(s).
Substituent groups useful for substituting saturated carbon atoms in the specified group or radical include, but are not limited to -Ra, halo, -O", =0, -ORb, -SRb, -S", =S, -NRCRC, =NRb, =N-ORb, trihalomethyl, -CF3, -CN, -OCN, -SCN, -NO, -NfY,, =N2, -N3, -S(0)2Rb, -S(0)2NRb, -S(0)2Cr, -S(0)2ORb, -OS(0)2Rb, -OS(0)20-, -OS(0)2ORb, -P(0)(0")2> -P(0)(ORb)(0-), -P(0)(ORb)(ORb), -C(0)Rb, -C(S)Rb, -C(NRb)Rb, -C(0)0", -C(0)ORb, -C(S)ORb, -C(0)NRcRc, -C(NRb)NRcRc, -OC(0)Rb, -OC(S)Rb, -OC(0)0", -OC(0)ORb, -OC(S)ORb, -NRbC(0)Rb, -NRbC(S)Rb, -NRbC(0)0", -NRbC(0)ORb, -NRbC(S)ORb, -NRbC(0)NRcRc, -NRbC(NRb)Rb and -NRbC(NRb)NRcRc, where Ra is selected from the group consisting of alkyl, cycloalkyl, . heteroalkyl, cycloheteroalkyl, aryl, arylalkyl, heteroaryl and heteroarylalkyl; each Rb is independently hydrogen or Ra; and each Rc is independently Rb or alternatively, the two R°s may be taken together with the nitrogen atom to which they are bonded form a 4-, 5-, 6- or
7-membered cycloheteroalkyl which may optionally include from 1 to 4 of the same or different additional heteroatoms selected from the group consisting of O, N and S. As specific examples, -NRCRC is meant to include -NH2, -NH-alkyl, N-pyrrolidinyl and N-morpholinyl. As another specific example, a substituted alkyl is meant to include -alkylene-O-alkyl, -alkylene-heteroaryl, -alkylene-cycloheteroalkyl, -alkylene-C(0)ORb, -alkylene-C(0)NRbRb, and -CH2-CH2-C(0)- C¾. The one or more substituent groups, taken together with the atoms to which they are bonded, may form a cyclic ring including cycloalkyl and cycloheteroalkyl.
Similarly, substituent groups useful for substituting unsaturated carbon atoms in the specified group or radical include, but are not limited to, -Ra, halo, -O", -ORb, -SRb, -S", -NRCRC, trihalomethyl, -CF3, -CN, -OCN, -SCN, -NO, -N02, -N3, -S(0)2Rb, -S(0)20\ -S(0)2ORb, -OS(0)2Rb, -OS(0)20", -OS(0)2ORb, -P(0)(Cr)2! -P(D)(ORb)(0"), -P(0)(ORb)(ORb), -C(0)Rb, -C(S)Rb, -C(NRb)Rb, -C(0)0", -C(0)ORb, -C(S)ORb, -C(0)NRcRc, -C(NRb)NRcRc, -OC(0)Rb, -OC(S)Rb, -OC(0)0", -OC(0)ORb, -OC(S)ORb, -NRbC(0)Rb, -NRbC(S)Rb, -NRbC(0)0-, -NRbC(0)ORb, -NRbC(S)ORb, -NRbC(0)NRcRc, -NRbC(NRb)Rb and -NRbC(NRb)NRcR°, where Ra, Rb and Rc are as previously defined.
Substituent groups useful for substituting nitrogen atoms in heteroalkyl and
cycloheteroalkyl groups include, but are not limited to, -Ra, -O", -ORb, -SRb, -S", -NRCRC, trihalomethyl, -CF3, -CN, -NO, -N02, -S(0)2Rb, -S(0)20-, -S(0)2ORb, -OS(0)2Rb, -OS(0)20\ -OS(0)2ORb, -P(0)(0")2, -P(0)(ORb)(0-), -P(0)(ORb)(ORb), -C(0)Rb, -C(S)Rb, -C(NRb)Rb, -C(0)ORb, -C(S)ORb, -C(0)NRcRc, -C(NRb)NR°Rc, -OC(0)Rb, -OC(S)Rb, -OC(0)ORb, -OC(S)ORb, -NRbC(0)Rb, -NRbC(S)Rb, -NRbC(0)ORb, -NRbC(S)ORb, -NRbC(0)NRcRc, -NRbC(NRb)Rb and -N RbC(N Rb)N RCRC, where Ra, Rb and Rc are as previously defined. Alkyl, alkenyl and alkynyl groups can alternatively or in addition be substituted by Cl - C8 acyl, C2-C8 heteroacyl, C6-C 10 aryl, C3-C8 cycloalkyl, C3-C8 heterocyclyl, or C5-C10 heteroaryl, each of which can be substituted by one or more R, halo, =0, =N-CN, =N-OR, =NR, OR, NR2, SR, S02R, S02NR2, NRS02R, NRCONR2, NRCSNR2, NRC(=NR)NR2, NRCOOR, NRCOR, CN, C≡CR, COOR, CONR2, OOCR, COR, and N02, wherein each R is independently H, C1 -C8 alkyl, C2-C8 heteroalkyl, C1 -C8 acyl, C2-C8 heteroacyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C3-C8 heterocyclyl, C4-C10
heterocyclylalkyl, C6-C10 aryl, or C5-C10 heteroaryl, and each R is optionally substituted with one or more (typically up to three) halo, =0, =N-CN, =N-OR', =NR', OR', NR'2, SR', S02R', S02NR'2, NR'S02R', NR'CONR'2, NR'CSNR'2, NR'C(=NR')NR'2, NR'COOR', NR'COR', CN, C≡CR', COOR', CONR'2, OOCR', COR', and N02, wherein each R' is independently H, C1 -C8 alkyl, C2-C8 heteroalkyl, C1 -C8 acyl, C3-C8 heterocyclyl, C2-C8 heteroacyl, C6-C 10 aryl or C5-C10 heteroaryl.
Where any of these substituents contains two R or R' groups on the same or adjacent atoms (e.g., -NR2, or -NR-C(O)R), the two R or R' groups can optionally be taken together with the atom(s) in the substituent group to which they are attached to form a ring having 5-8 ring members, which can include another heteroatom as a ring member (N, O or S) and can be substituted with one or more halo, =0, =N-CN, =N-OR, =NR, OR, NR2, SR, S02R, S02NR2, . NRS02R, NRCONR2, NRCSNR2, NRC(=NR)NR2, NRCOOR, NRCOR, CN, C≡CR, COOR, CONR2, OOCR, COR, and N02, wherein each R is independently H, C1 -C8 alkyl, C2-C8 heteroalkyl, C1 -C8 acyl, C2-C8 heteroacyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C3-C8 heterocyclyl, C4-C10 heterocyclylalkyl, C6-C10 aryl, or C5-C10 heteroaryl, and each R is optionally substituted with halo, =0, =N-CN, =N-OR', =NR\ OR', NR'2, SR', S02R', S02NR'2, NR'S02R', NR'CONR'2, NR'CSNR'2, NR'C(=NR')NR'2, NR'COOR', NR'COR', CN, C≡CR', COOR', CONR'2, OOCR', COR', and N02, wherein each R' is independently H, C1 -C8 alkyl, C2-C8 heteroalkyl, C1 -C8 acyl, C3-C8 heterocyclyl, C2-C8 heteroacyl, C6-C 10 aryl or C5-C10 heteroaryl, and each of the substitutable groups on R' can be substituted with one or more (e.g., up to three) halo, piperidinyl, pyrrolidinyl, piperazinyl, morpholinyl, CN, C1 -C4 alkoxy, OH, OAc, NH2, C 1 -C4 alkyl amine, di(Cl -C4 alkyl)amine, NHAc, NHCOOMe, NHCOOEt, NHCOOtBu, NHS02Me, SMe, S02Me, S02NH2, S02NMe2, COOH, CONH2, COOMe, COOEt, CONHMe, or CONM^ "Acetylene" substituents are 2-1 OC alkynyl groups that contain at least one carbon- carbon triple bond and are optionally substituted with the groups described herein as suitable for alkyl groups; in some embodiments, the alkynyl groups are of the formula -C≡C-Ra, wherein Ra is H or C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C1 -C8 acyl, C2-C8 heteroacyi, C6-C10 aryl, C5-C10 heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkyl.
Each Ra group is optionally substituted with one or more substituents selected from halo, =0, =N-CN, =N-OR', =NR\ OR', NR'2, SR', S02R\ S02NR'2, NR'S02R\ NR'CONR'2, NR'CSNR'2, NR'C(=NR')NR'2, NR'COOR', NR'COR', CN, COOR', CONR'2) OOCR', COR', and N02, wherein each R' is independently H, C1-C6 alkyl, C2-C6 heteroalkyl, C1-C6 acyl, C2-C6 heteroacyi, C6-C10 aryl, C5-C10 heteroaryl, C7-12 arylalkyl, or C6-12
heteroarylalkyl, each of which is optionally substituted with one or more groups selected from halo, CN, C1 -C4 alkyl, C2-C4 heteroalkyl, C1-C6 acyl, C1 -C6 heteroacyi, C1-C4 alkoxy, C1-C4 alkylamino, di(Cl -C4 alkyl)amino, hydroxy, amino, and =0; and wherein two R' can be linked to form a 3-7 membered ring optionally containing up to three heteroatoms selected from N, O and S. In some embodiments, Ra of -C≡C-Ra is H or Me.
"Heteroalkyl", "heteroalkenyl", and "heteroalkynyl" and the like are defined similarly to the corresponding hydrocarbyl (alkyl, alkenyl and alkynyl) groups, but the 'hetero' terms refer to groups that contain 1 -3 O, S or N heteroatoms or combinations thereof within the backbone residue; thus at least one carbon atom of a corresponding alkyl, alkenyl, or alkynyl group is replaced by one of the specified heteroatoms to form, respectively, a heteroalkyl, heteroalkenyl, or heteroalkynyl group. The typical and preferred sizes for heteroforms of alkyl, alkenyl and alkynyl groups are generally the same as for the corresponding hydrocarbyl groups, and the substituents that may be present on the heteroforms are the same as those described above for the hydrocarbyl groups. For reasons of chemical stability, it is also understood that, unless otherwise specified, such groups do not include more than two contiguous heteroatoms except where an oxo group is present on N or S as in a nitro or sulfonyl group.
While "alkyl" as used herein includes cycloalkyl and cycloalkylalkyl groups, the term "cycloalkyl" may be used herein to describe a carbocyclic non-aromatic group that is connected via a ring carbon atom, and "cycloalkylalkyl" may be used to describe a carbocyclic non- aromatic group that is connected to the molecule through alkyl linker. Similarly, "heterocyclyl" may be used to describe a non-aromatic cyclic group that contains at least one heteroatom (typically selected from N, O and S) as a ring member and that is connected to the molecule via a ring atom, which may be C (carbon-linked) or N (nitrogen- linked); and "heterocyclylalkyl" may be used to describe such a group that is connected to another molecule through a linker. The heterocyclyl can be fully saturated or partially saturated, but non-aromatic. The sizes and substituents that are suitable for the cycloalkyl, cycloalkylalkyl, heterocyclyl, and heterocyclylalkyl groups are the same as those described above for alkyl groups. The heterocyclyl groups typically contain 1 , 2 or 3 heteroatoms, selected from N, O and S as ring members; and the N or S can be substituted with the groups commonly found on these atoms in heterocyclic systems. As used herein, these terms also include rings that contain a double bond or two, as long as the ring that is attached is not aromatic. The substituted cycloalkyl and heterocyclyl groups also include cycloalkyl or heterocyclic rings fused to an aromatic ring or heteroaromatic ring, provided the point of attachment of the group is to the cycloalkyl or heterocyclyl ring rather than to the aromatic / heteroaromatic ring.
Like alkyl groups, the cycloalkyl and heterocyclyl groups described herein can be substituted to the extent permitted by their valence and stability considerations, which are well understood by those of skill in the art. Substituents for the cycloalkyl and heterocyclyl rings or ring systems include those described herein as suitable for placement on alkyl groups.
Any undefined valency on an atom of a structure shown in this application implicitly represents a hydrogen atom bonded to the atom. As used herein, "acyl" encompasses groups comprising an alkyl, alkenyl, alkynyl, aryl or arylalkyl radical attached at one of the two available valence positions of a carbonyl carbon atom, and heteroacyl refers to the corresponding groups wherein at least one carbon other than the carbonyl carbon has been replaced by a heteroatom chosen from N, O and S. Thus heteroacyl includes, for example, -C(=0)OR and - C(=0)NR2 as well as -C(=0)-heteroaryl.
Acyl and heteroacyl groups are bonded to any group or molecule to which they are attached through the open valence of the carbonyl carbon atom. Typically, they are C1 -C8 acyl groups, which include formyl, acetyl, pivaloyl, and benzoyl, and C2-C8 heteroacyl groups, which include methoxyacetyl, ethoxycarbonyl, and 4-pyridinoyl. The hydrocarbyl groups, aryl groups, and heteroforms of such groups that comprise an acyl or heteroacyl group can be substituted with the substituents described herein as generally suitable substituents for each of the corresponding component of the acyl or heteroacyl group.
"Aromatic" moiety or "aryl" moiety refers to a monocyclic or fused bicyclic moiety having the well-known characteristics of aromaticity; examples include phenyl and naphthyl. Similarly, "heteroaromatic" and "heteroaryl" refer to such monocyclic or fused bicyclic ring systems which contain as ring members one or more heteroatoms selected from O, S and N. The inclusion of a heteroatom permits aromaticity in 5-membered rings as well as 6-membered rings. Typical heteroaromatic systems include monocyclic C5-C6 aromatic groups such as pyridyl, pyrimidyl, pyrazinyl, thienyl, furanyi, pyrrolyi, pyrazoiyl, thiazolyi, oxazoiyl, triazolyl, triazinyl, tetrazolyl, tetrazinyl, and imidazolyl and the fused bicyclic moieties formed by fusing one of these monocyclic groups with a phenyl ring or with any of the heteroaromatic monocyclic groups to form a C8-C 10 bicyclic group such as indolyl, benzimidazolyl, indazolyl, benzotriazolyl, isoquinolyl, quinolyl, benzothiazolyl, benzofuranyl, pyrazolopyridyl, quinazolinyl, quinoxalinyl, cinnolinyl, and the like. Any monocyclic or fused ring bicyclic system which has the characteristics of aromaticity in terms of electron distribution throughout the ring system is included in this definition. It also includes bicyclic groups where at least the ring which is directly attached to the remainder of the molecule has the characteristics of aromaticity.
Typically, the ring systems contain 5-12 ring member atoms and up to four heteroatoms selected from N, O and S. Frequently, the monocyclic heteroaryls contain 5-6 ring members and up to three such heteroatoms, and the bicyclic heteroaryls contain 8- 10 ring members and up to four such heteroatoms. The number and placement of heteroatoms in such rings is in accordance with the well-known limitations of aromaticity and stability, where stability requires the
heteroaromatic group to be stable enough to be exposed to water without rapid degradation.
Aryl and heteroaryl moieties may be substituted with a variety of substituents including C1 -C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C5-C12 aryl, C1 -C8 acyl, and heteroforms of these, each of which can itself be further substituted; other substituents for aryl and heteroaryl moieties include halo, OR, NR2, SR, S02R, S02NR2, NRS02R, NRCONR2, NRCSNR2, NRC(=NR)NR2, NRCOOR, NRCOR, CN, OCR, COOR, CONR2, OOCR, COR, and N02, wherein each R is independently H, C1 -C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C3-C8 heterocyclyl, C4-C 10 heterocyclylalkyl, C6-C 10 aryl, C5- C10 heteroaryl, C7-C12 arylalkyl, or C6-C 12 heteroaryla'kvl, and each R is optionally substituted as described above for alkyl groups. The substituent groups on an aryl or heteroaryl group may of course be further substituted with the groups described herein as suitable for each type of such substituents or for each component of the substituent. Thus, for example, an arylalkyl substituent may be substituted on the aryl portion with substituents described herein as typical for aryl groups, and it may be further substituted on the alkyl portion with substituents described herein as typical or suitable for alkyl groups. Where a substituent group contains two R groups on the same or adjacent atoms (e.g., -NR2, or -NR-C(O)R), the two R groups can optionally be taken together with the atom(s) in the substituent group to which the are attached to form a ring having 5-8 ring members, which can be substituted as allowed for the R itself, and can contain an additional heteroatom (N, O or S) as a ring member.
Similarly, "arylalkyl" and "heteroarylalkyl" refer to aromatic and heteroaromatic ring systems which are bonded to their attachment point through a linking group such as an alkylene, including substituted or unsubstituted, saturated or unsaturated, cyclic or acyclic linkers.
Typically the linker is C1 -C8 alkyl or a hetero form thereof. These linkers may also include a carbonyl group, thus making them able to provide substituents as an acyl or heteroacyl moiety. An aryl or heteroaryl ring in an arylalkyl or heteroarylalkyl group may be substituted with the same substituents described above for aryl groups. Preferably, an arylalkyl group includes a phenyl ring optionally substituted with the groups defined above for aryl groups and a C 1 -C4 alkylene that is unsubstituted or is substituted with one or two C1 -C4 alkyl groups or heteroalkyl groups, where the alkyl or heteroalkyl groups can optionally cyclize to form a ring such as cyclopropane, dioxolane, or oxacyclopentane. Similarly, a heteroarylalkyl group preferably includes a C5-C6 monocyclic heteroaryl group that is optionally substituted with the groups described above as substituents typical on aryl groups and a C1 -C4 alkylene that is unsubstituted or is substituted with one or two C 1 -C4 alkyl groups or heteroalkyl groups, or it includes an optionally substituted phenyl ring or C5-C6 monocyclic heteroaryl and a C1 -C4 heteroalkylene that is unsubstituted or is substituted with one or two C 1 -C4 alkyl or heteroalkyl groups, where the alkyl or heteroalkyl groups can optionally cyclize to form a ring such as cyclopropane, dioxolane, or oxacyclopentane.
Where an arylalkyl or heteroarylalkyl group is described as optionally substituted, the substituents may be on either the alkyl or heteroalkyl portion or on the aryl or heteroaryl portion of the group. The substituents optionally present on the a'^vl or heteroalkyl portion are the same as those described above for alkyl groups generally; the substituents optionally present on the aryl or heteroaryl portion are the same as those described above for aryl groups generally.
"Arylalkyl" groups as used herein are hydrocarbyl groups if they are unsubstituted, and are described by the total number of carbon atoms in the ring and alkylene or similar linker. Thus a benzyl group is a C7-arylalkyl group, and phenylethyl is a C8-arylalkyl.
"Heteroarylalkyl" as described above refers to a moiety comprising an aryl group that is attached through a linking group, and differs from "arylalkyl" in that at least one ring atom of the aryl moiety or one atom in the linking group is a heteroatom selected from N, O and S. The heteroarylalkyl groups are described herein according to the total number of atoms in the ring and linker combined, and they include aryl groups linked through a heteroalkyl linker; heteroaryl groups linked through a hydrocarbyl linker such as an alkylene; and heteroaryl groups linked through a heteroalkyl linker. Thus, for example, C7-heteroarylalkyl would include
pyridylmethyl, phenoxy, and N-pyrrolylmethoxy.
"Alkylene" as used herein refers to a divalent hydrocarbyl group; because it is divalent, it can link two other groups together. Typically it refers to -{CH2)n- where n is 1 -8 and preferably n is 1 -4, though where specified, an alkylene can also be substituted by other groups, and can be of other lengths, and the open valences need not be at opposite ends of a chain. Thus -CH(Me)- and -C(Me)2- may also be referred to as alkylenes, as can a cyclic group such as
cyclopropan-l , l -diyl. Where an alkylene group is substituted, the substituents include those typically present on alkyl groups as described herein.
In general, any alkyl, alkenyl, alkynyl, acyl, or aryl or arylalkyl group or any heteroform of one of these groups that is contained in a substituent may itself optionally be substituted by additional substituents. The nature of these substituents is similar to those recited with regard to the primary substituents themselves if the substituents are not otherwise described. Thus, where an embodiment of, for example, R" is alkyl, this alkyl may optionally be substituted by the remaining substituents listed as embodiments for Rx where this makes chemical sense, and where this does not undermine the size limit provided for the alkyl per se; e.g. , alkyl substituted by alkyl or by alkenyl would simply extend the upper limit of carbon atoms for these embodiments, and is not included. However, alkyl substituted by aryl, amino, alkoxy, =0, and the like would be included within the scope of the invention, and the atoms of these substituent groups are not counted in the number used to describe the alkyl, alkenyl etr.. group that is being described. Where no number of substituents is specified, each such alkyl, alkenyl, alkynyl, acyl, or aryl group may be substituted with a number of substituents according to its available valences; in particular, any of these groups may be substituted with fluorine atoms at any or all of its available valences, for example.
"Heteroform" as used herein refers to a derivative of a group such as an alkyl, aryl, or acyl, wherein at least one carbon atom of the designated carbocyclic group has been replaced by a heteroatom selected from N, O and S. Thus the heteroforms of alkyl, alkenyl, alkynyl, acyl, aryl, and arylalkyl are heteroalkyl, heteroalkenyl, heteroalkynyl, heteroacyl, heteroaryl, and heteroarylalkyl, respectively. It is understood that no more than two N, O or S atoms are ordinarily connected sequentially, except where an oxo group is attached to N or S to form a nitro or sulfonyl group.
"Halo", as used herein includes fluoro, chloro, bromo and iodo. Fluoro and chloro are often preferred.
"Amino" as used herein refers to NH2, but where an amino is described as "substituted" or "optionally substituted", the term includes NR'R" wherein each R' and R" is independently H, or is an alkyl, alkenyl, alkynyl, acyl, aryl, or arylalkyl group or a heteroform of one of these groups, and each of the alkyl, alkenyl, alkynyl, acyl, aryl, or arylalkyl groups or heteroforms of one of these groups is optionally substituted with the substituents described herein as suitable for the corresponding group. The term also includes forms wherein R' and R" are taken together with the N to which they are attached to form a 3-8 membered ring which may be saturated, unsaturated or aromatic and which contains 1 -3 heteroatoms independently selected from N, O and S as ring members, and which is optionally substituted with the substituents described as suitable for alkyl groups or, if NR'R" is an aromatic group, it is optionally substituted with the substituents described as typical for heteroaryl groups.
As used herein, the term "carbocycle", "carbocyclyl", or "carbocyclic" refers to a cyclic ring containing only carbon atoms in the ring, whereas the term "heterocycle" or "heterocyclic" refers to a ring comprising a heteroatom. The carbocyclyl can be fully saturated or partially saturated, but non-aromatic. For example, the carbocyclyl encompasses cycloalkyl. The carbocyclic and heterocyclic structures encompass compounds having monocyclic, bicyclic or multiple ring systems; and such systems may mix aromatic, heterocyclic, and carbocyclic rings.
Mixed ring systems are described according to the ring that is attached to the rest of the compound being described; for example, where W represents 1 ,2,3,4-tetrahydronaphth-l -yl, the group would be encompassed by an optionally substituted cycloalkyl or carbocyclic group, while the group l ,2,3,4-tetrahydronaphth-6-yl would be included within optionally substituted aromatic groups.
As used herein, the term "heteroatom" refers to any atom that is not carbon or hydrogen, such as nitrogen, oxygen or sulfur. When it is part of the backbone or skeleton of a chain or ring, a heteroatom must be at least divalent, and will typically be selected from N, O, P, and S.
Illustrative examples of heterocycles and heteroaryls include but are not limited to tetrahydrofuran, 1 ,3-dioxolane, 2,3-dihydrofuran, pyran, tetrahydropyran, benzofuran, isobenzofuran, 1 ,3-dihydro-isobenzofuran, isoxazole, 4,5-dihydroisoxazole, piperidine, pyrrolidine,- pyrrolidin-2-one, pyrrole, pyridine, pyrimidine, octahydro-pyrrolo[3,4-Z>]pyridine, piperazine, pyrazine, morpholine, thiomorpholine, imidazole, imidazolidine 2,4-dione, 1 ,3- dihydrobenzimidazol-2-one, indole, thiazole, benzothiazole, thiadiazole, thiophene, tetrahydro thiophene 1 , 1 -dioxide, diazepine, triazole, guanidine, diazabicyclo[2.2.1]heptane, 2,5- diazabicyclo[2.2.1 ]heptane, 2,3,4,4a,9,9a-hexahydro- l H-P-carboline, oxirane, oxetane, tetrahydropyran, dioxane, lactones, aziridine, azetidine, piperidine, lactams, and may also encompass heteroaryls. Other illustrative examples of heteroaryls include but are not limited to furan, pyrrole, pyridine, pyrimidine, imidazole, benzimidazole and triazole.
Embodiments of the Compounds:
In one embodiment, the present invention provides compounds having the general formula (1):
Figure imgf000025_0001
or a pharmaceutically acceptable salt, solvate, and/or prodrug thereof; wherein:
Z4 is N or CR5;
R5 is halo, -CN, -R,-OR, (0)nR, -COOR, -CONR2, -NR2, optionally substituted aryl, or optionally substituted heteroaryl;
R2, R3 and R4 are each independently H or optionally substituted CI -CIO alkyl;
X is O, S, or NR6;
Y is O or S or NR10;
R6 and R10 are each independently selected from H, CN, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C1-C4 alkoxy, optionally substituted C6-C10 aryl, optionally substituted heteroaryl, and NR7R8;
Z is O or S;
L is a covalent bond, -NR7-, -0-, -S(0)n-, -(CR7R8)m-NR9-, -(CR7R8)m-0-, or -(CR7R8)m-
S(0)„-;
W is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, or optionally substituted carbocyclyl;
where each R7 and R8 and R9 is independently selected from H, optionally substituted CI -CIO alkyl, optionally substituted heteroalkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl;
or R7 and R8, taken together on a single carbon atom or on adjacent connected carbon atoms of (CR7R8)m whether alone or as part of another group, form a 3 to 8 membered carbocyclic ring or heterocyclic ring;
or R7 and R8, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 to 10 membered heterocyclic or heteroaryl ring that optionally contains one or more additional heteroatom selected from N, O and S as a ring member;
each m is independently 1 , 2, 3 or 4;
G is NR1ARIB, OR1A, or SR1A;
R1A is H or optionally substituted C1-C10 alkyl; RIB is H, optionally substituted C I -C I O alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, or optionally substituted heteroarylalkyl; each R is independently H or optionally substituted C 1 -C4 alkyl; or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member; and
each n is independently 0, 1 , or 2.
In any one of the preceding embodiments of Formula (I), Z4 is N or CH.
In any one of the preceding embodiments of Formula (I), R3 and R4 are both H.
In any one of the preceding embodiments of Formula (I), R2 is H, -CH3, halo, -OCH3, or
-CF3.
In any one of the preceding embodiments of Formula (I), Y is O or S.
In any one of the preceding embodiments of Formula (I), Z is O.
In any one of the preceding embodiments of Formula (I), X is selected from the group consisting of NH, O, and S.
In any one of the preceding embodiments of Formula (I), L is a covalent bond; and W is optionally substituted aryl, optionally substituted heteroaryl, or optionally substituted heterocyclyl.
In any one of the preceding embodiments of Formula (I), L is -NR7-, -0-, -S(0)n-, -(CR7R8)m-NR9-, -(CR7R8)m-0-, or -(CR7R8)m-S(0)n-; R7 and R8 and R9 are each independently H or C1 -C 10 alkyl; and W is optionally substituted aryl or optionally substituted carbocyclyl.
In any one of the preceding embodiments of Formula (I), W-L- is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, (optionally substituted aryl)-NR7-, (optionally substituted heteroaryl)-NR7-, (optionally substituted aryl)-CH2-NR7-, (optionally substituted heteroaryl)-CH2-NR7-, (optionally substituted aryl)-0-, (optionally substituted heteroaryl)-0-, (optionally substituted aryl)-S-, (optionally substituted heteroaryl)-S-, (optionally substituted aryl)-CH2-0-, or (optionally substituted heteroaryl)-CH2-0-; and R7 is each independently H or C I -C I O alkyl. In one embodiment of Formula (I), Z4 is N or CR5; R5 is -R, -CN, -OR, -Ar, COOR, - NR2; Z and Y are both O; R2 and R4 are both H; R3 is H, optionally substituted CI -CI O alkyl, or optionally substituted C1 -C 10 heteroalkyl; X is NH; RIB is optionally substituted
carbocyclylalkyl; L is a covalent bond, -NR7-, or -CH2-NR7-; R7 is H or C 1 -C4 alkyl; and W is optionally substituted aryl or optionally substituted heteroaryl. In one embodiment of W, the one or more substituents are selected from the group consisting of optionally substituted CI -CI O alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted CI -C I O heteroalkyl, -OR, halo, -S(0)R, -S(0)2R, -CN, -NR2, -SR, -N02, -C(0)R, - C(0)OR, -NHC(0)R, -NRC(0)R, -C(0)NR2, -CF3, optionally substituted aryl, and optionally substituted heteroaryl; and each R is independently selected from H and and optionally substituted C 1-C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member, and wherein Ar is optionally substituted aryl or optionally substituted heteroaryl. In another embodiment of W, the one or more substitutents are selected from the group consisting of phenyl, thiophene, pyridine, pyrimidine, pyrazine, pyridazine, pyrrole, furan, thiofuran, oxazole, isooxazole, thiazole, quinoline, isoquinoline, indole, and pyrazole, wherein each substitutent is optionally substituted with one or more substitutents selected from the group consisting of optionally substituted C1 -C10 alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted CI -C I O heteroalkyl, -OR, halo, -S(0)R, -S(0)2R, -CN, -NR2, -SR, -N02, -C(0)R, -C(0)OR, -NHC(0)R, -NRC(0)R, -C(0)NR2, -CF3, optionally substituted aryl, and optionally substituted heteroaryl; and each R is independently selected from H and and optionally substituted C 1 -C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member, and wherein Ar is optionally substituted aryl or optionally substituted heteroaryl. In yet another embodiment of W, the one or more substitutents are selected from the group consisting of aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine, tetrahydrofuran, pyran, thiopyran, thiomorpholine, thiomorpholine S-oxide, thiomorpholine S-dioxide, oxazoline, tetrahydrothiophene, piperidine, tetrahydropyran, thiane, imidazolidine, oxazolidine, thiazolidine, dioxolane, dithiolane, piperazine, oxazine, dithiane, morpholine, and dioxane, wherein each substitutent is optionally substituted with one or more substitutents selected from the group consisting of optionally substituted C1 -C10 alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted Cl - C10 heteroalkyl, -OR, halo, -S(0)R, -S(0)2R, -CN, -NR2, -SR, -N02, -C(0)R, -C(0)OR, - NHC(0)R, -NRC(0)R, -C(0)NR2, -CF3, optionally substituted aryl, and optionally substituted heteroaryl; and each R is independently selected from H and and optionally substituted C1 -C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member, and wherein Ar is optionally substituted aryl or optionally substituted heteroaryl. In yet another embodiment of W, the one or more substitutents are selected from the group consisting of CI, Br, F, -OCF3, -CF3, -CH3, -C2H5, -C3H7, -OCH20-, OCH2CH20-, -SCH3, -OCH3, -NHCOCH3, - NH2, -COCH3, -N02, -0(CH2)i.6-COOR, and -S02CH3. In any one of the preceding embodiments, W can be an optionally substituted phenyl or an optionally substituted pyridine.
In one embo ula (II):
Figure imgf000029_0001
wherein:
p is 0, 1 , 2 or 3;
Z4 is N or CR5;
R5 is H or C l -C4 alkyl;
is H or S;
R3 is H, C1 -C 10 alkyl, or C 1 -C10 heteroalkyl;
R7 is H or C l -C4 alkyl; Ar is aryl or heteroaryl which is optionally substituted by one or more substituents selected from the group consisting of optionally substituted C I -C I O alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted C2-C 10 alkenyl, optionally substituted C2-C10 alkynyl, -NR2; -OR, S(0)nR; halo, -CN, -N02, -C(0)OR, - C(0)NR2, or -C(0)R;
n is 0, 1 , or 2;
each R is independently H or optionally substituted C 1 -C4 alkyl; or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member; and
RIB is selected from H, optionally substituted C I -C I O alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylaikyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, or optionally substituted heteroarylalkyl.
In one embodiment of Formula (II), Ar is optionally substituted phenyl or pyridinyl. In one embodiment of Formula (II), p is 0 or 1 .
In one embodiment of Formula (II), the compound has structural Formula (Ila), (lib), (H
Figure imgf000030_0001
wherein:
Z4 is N or CR5;
R5 is H or C l -C4 alk l;
X is NH, or S;
R3 is H, C1 -C10 alkyl or Cl -CI O heteroalkyl;
R7 is H or C l -C4 alkyl;
q is 0, 1 , 2, 3, 4, or 5;
t is 0, 1 , 2, 3, or 4;
A is optionally substituted C1 -C10 alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted C2-C 10 alkenyl, optionally substituted C2-C10 alkynyl, -NR2; -OR, S(0)nR; halo, -CN, -N02, -C(0)OR, -C(0)NR2, or -C(0)R;
n is 0, 1 , or 2;
each R is independently H or optionally substituted C 1 -C4 alkyl; or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member; and
R1B is cyclopropyl or cyclopropylmethylene.
In one embodiment of Formula (11a), (lib), (lie) or (lid), A is selected from the group consisting of phenyl, thiophene, pyridine, pyrimidine, pyrazine, pyridazine, pyrrole, furan, thiofuran, oxazole, isooxazole, thiazole, quinoline, isoquinoline, indole, and pyrazole, each of which is optionally substituted with one or more substitutents selected from the group consisting of optionally substituted C 1 -C10 alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted C I -C I O heteroalkyl, -OR, halo, -S(0)R, -S(0)2R, -CN, -NR2, -SR, -N02, -C(0)R, -C(0)OR, -NHC(0)R, -NRC(0)R, -C(0)NR2, -CF3, optionally substituted aryl, and optionally substituted heteroaryl; and each R is independently selected from H and and optionally substituted C I -C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member, and wherein Ar is optionally substituted aryl or optionally substituted heteroaryl. In one embodiment of Formula (lla), (lib), (lie) or (lid), A is selected from the group consisting of aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine,
tetrahydrofuran, pyran, thiopyran, thiomorpholine, thiomqrpholine S-oxide, thiomorpholine S- dioxide, oxazoline, tetrahydrothiophene, piperidine, tetrahydropyran, thiane, imidazolidine, oxazolidine, thiazolidine, dioxolane, dithiolane, piperazine, oxazine, dithiane, morpholine, and dioxane, each of which is optionally substituted with one or more substitutents selected from the group consisting of optionally substituted CI -C I O alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted CI -C I O heteroalkyl, -OR, halo, - S(0)R, -S(0)2R, -CN, -NR2, -SR, -N02, -C(0)R, -C(0)OR, -NHC(0)R, -NRC(0)R, -C(0)NR2, -CF3, optionally substituted aryl, and optionally substituted heteroaryl; and each R is independently selected from H and and optionally substituted C1 -C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member, and wherein Ar is optionally substituted aryl or optionally substituted heteroaryl.
In one embodiment of Formula (Ha), (lib), (lie) or (lid), A is selected from the group consisting of CI, Br, F, -OCF3, -CF3, -CH3, -C2H5, -C3H7, -OCH20-, OCH2CH20-, -SCH3, - OCH3, -NHCOCH3, -NH2, -COCH3, -N02, -0(CH2)1-6-COOR, and -S02CH3.
In one embodiment of Formula (I), the compound has a structural Formula (III):
Figure imgf000032_0001
wherein:
Z4 is N or CR5;
R5 is halo, -CN, -R,-OR, -S(0)nR, -COOR, -CONR2, -NR2, optionally substituted aryl, or optionally substituted heteroaryl; R2, R3 and R4 are each independently is H or optionally substituted C 1 -C10 alkyl;
X is O, S, or R6;
Y is O, S, or NR10;
R6 and R10 are each independently selected from H, CN, optionally substituted C 1 -C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C1 -C4 alkoxy, optionally substituted C6-C 10 aryl, and optionally substituted heteroaryl;
Z is O or S;
hCy is an optionally substituted 5 to 10 membered heterocyclic or heteroaryl ring that contains one or more heteroatom selected from N, O and S as a ring member;
G is NR1AR1B, OR1A, or SR1A;
R,A is H or optionally substituted C1 -C 10 alkyl;
RIB is selected from H, optionally substituted C I -CI O alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, or optionally substituted heteroarylalkyl;
each R is independently H or optionally substituted C 1 -C4 alkyl; or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member; and
n is 0, 1 , or 2.
In one embodiment of Formula (III), hCy is selected from the group consisting of thiophene, pyridine, pyrimidine, pyrazine, pyridazine, pyrrole, furan, thiofuran, oxazole, isooxazole, thiazole, quinoline, isoquinoline, indole, and pyrazole, each of which is optionally substituted with one or more substitutents selected from the group consisting of optionally substituted C I -C I O alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted C I -C I O heteroalkyl, -OR, halo, -S(0)R, -S(0)2R, -CN, -NR2, -SR, -N02, - C(0)R, -C(0)OR, -NHC(0)R, -NRC(0)R, -C(0)NR2, -CF3, optionally substituted aryl, and optionally substituted heteroaryl; and each R is independently selected from H and and optionally substituted C1 -C4 alkyl, or alternatively, the tw groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, 0, and S as a ring member, and wherein Ar is optionally substituted aryl or optionally substituted heteroaryl.
In one embodiment of Formula (III), the compound has a structural Formula (Ilia):
Figure imgf000034_0001
wherein:
each A is independently optionally substituted C I -CI O alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted C2-C10 alkenyl, optionally substituted C2-C 10 alkynyl, -NR2; -OR, S(0)nR; halo, -CN, -N02, -C(0)OR, - C(0)NR2, or -C(0)R;
c is 0, 1 , 2, or 3;
n is 0, 1 , or 2; and
each R is independently H or optionally substituted C 1 -C4 alkyl; or alternatively, the two
R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member.
In one embodiment of Formula (Ilia), Z4 is CR5; R2 and R4 are H; Z and Y are O; X is NH or S; G is NRIARI B; R1A is H or methyl; R1B is optionally substituted C I -C I O alkyl, optionally substituted CI -CI O cylcoalkyl, optionally substituted heterocyclyl, optionally substituted C 1 -C 10 alkylether, optionally substituted C1 -C 10 alkylaminoalkyl, optionally substituted C I -C I O alkoxyalkyl, optionally substituted heterocyclylalkyl, alkylaryl, aralkyl, alkylheteroaryl, heteroarylalkyl, or optionally substituted aryl. In one embodiment of Formula (Ilia), R1B is cyclopropyl, cyclopropylmethylene, cyclobutyl, cyclohexyl, optionally substituted morpholinylalkyl, or optionally substituted phenyl.
In any one of preceding embodiments of Formula (Ilia), A is -CN, C 1 -C4 alkyl, - C(0)OR, or -C(0)NR2.
In one embodiment of Formula (III), hCy is selected from the group consisting of aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine, tetrahydrofuran, pyran, thiopyran, thiomorpholine, thiomorpholine S-oxide, thiomorpholine S-dioxide, oxazoline, tetrahydrothiophene, piperidine, tetrahydropyran, thiane, imidazolidine, oxazolidine, thiazolidine, dioxolane, dithiolane, piperazine, oxazine, dithiane, morpholine, and dioxane, each of which is optionally substituted with one or more substitutents selected from the group consisting of optionally substituted C1 -C10 alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted CI -C IO heteroalkyl, -OR, halo, -S(0)R, -S(0)2R, -CN, -NR2, -SR, -N02; -C(0)R, -C(0)0R, -NHC(0)R, -NRC(0)R, -C(0)NR2, -CF3, optionally substituted aryl, and optionally substituted heteroaryl; and each R is independently selected from H and and optionally substituted C1 -C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member, and wherein Ar is optionally substituted aryl or optionally substituted heteroaryl.
In one embodiment of Formula (III), the compound has a structural Formula (Illb):
Figure imgf000035_0001
wherein
D is N or CH; a and b are independently 1 , 2, or 3;
Z4 is 1M or CR5;
R5 is halo, -CN, -R,-OR, -S(0)nR, -COOR, -CONR2, -NR2, optionally substituted aryl, or optionally substituted heteroaryl;
R2, R3 and R4 are each independently selected from H and optionally substituted CI -C I O alkyl;
X is O, S, or NR6;
Y is O or S or NR10;
Z is O or S;
R6 and R10 are each independently selected from H, CN, optionally substituted C 1 -C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C1 -C4 alkoxy, optionally substituted C6-C 10 aryl, and optionally substituted heteroaryl;
L2 is optionally substituted CI -CIO alkyl, optionally substituted CI -CIO heteroalkyl, optionally substituted C2-C 10 alkenyl, optionally substituted C2-C10 alkynyl, optionally substituted aryl, optionally substituted heteroaryl, -C(O)-, -S(0)n-, or -CR2-;
E is -OR, -NR, optionally substituted C 1 -C10 alkyl, optionally substituted C1 -C 10 heteroalkyl, optionally substituted aryl, or optionally substituted heteroaryl;
RIB is H, optionally substituted C I -C I O alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, or optionally substituted heteroarylalkyl; and
each R is independently selected from H and optionally substituted C 1 -C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member; and
n is 0, 1 , or 2.
In one embodiment of Formula (Illb), Z4 is N or CH; R2', R3, and R4 are H; Z and Y are O; X is NH; and RI B is cyclopropyl or cyclopropylmethylene.
In one embodiment of Formula (Illb), a and b are hr>th 2. In any one of the preceding embodiments of Formula (111b), L2 is -C(O)- or -S(0)2-; and E is optionally substituted aryl or optionally substituted heteroaryl.
In any one of the preceding embodiments of Formula (Illb), L2 is -CH2-; and E is optionally substituted C 1 -C4 alkyl, optionally substituted aryl, or optionally substituted heteroaryl.
In any one of the preceding embodiments of Formula (Illb), the optionally substituted aryl is optionally substituted phenyl, and the optionally substituted heteroaryl is optionally substituted pyridyl.
In any one of the preceding embodiments of Formula (Illb), the optionally substituted aryl or the optionally substituted heteroaryl is independently optionally substituted by one or more substituents selected from the group consisting of optionally substituted C l -CIO alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted Cl - CI O heteroalkyl, -OR, halo, -S(0)R, -S(0)2R, -CN, -NR2) -SR, -N02, -C(0)R, -C(0)OR, - NHC(0)R, -NRC(0)R, -C(0)NR2, optionally substituted aryl, and optionally substituted heteroaryl; and each R is independently selected from H and and optionally substituted C1 -C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member.
In one embo Formula (IV):
Figure imgf000037_0001
V), wherein:
L1 is O or S;
L3 is a covalent bond or an optionally substituted C 1 -C4 alkylene; Z4 each independently represent N or CR5;
R5 is halo, -CN, -R,-OR, -S(0)nR, -COOR, -CONR2, -NR2, optionally substituted aryl, or optionally substituted heteroaryl;
each R is independently selected from H and optionally substituted C 1 -C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member;
n is 0, 1 , or 2;
R2, R3 and R4 are each independently selected from H and optionally substituted CI -CI O alkyl;
X is O, S, or NR6;
Y is O or S or NR10;
R6 and R10 are each independently selected from H, CN, optionally substituted C 1 -C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C 1 -C4 alkoxy, optionally substituted C6-C 10 aryl, and optionally substituted heteroaryl;
Z is O or S;
Ar is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, or optionally substituted heterocyclyl; and
RIB is selected from H, optionally substituted C I -CI O alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl.
In one embodiment of Formula (IV), Z4 is CH; R2, R3, and R4 are H; Z and Y are O; X is
NH; R1 is cyclopropyl; L3 is a covalent bond or an optionally substituted C1 -C4 alkylene; and Ar is optionally substituted aryl or heteroaryl.
In one embodiment of of Formula (IV), L3 is a covalent bond or methylene.
In any one of the preceding embodiments of Formula (IV), Ar is optionally substituted phenyl, optionally substituted pyridyl, or optionally substituted indolyl.
In one embodiment of Formula (I), the compound has a structural Formula (V):
Figure imgf000039_0001
wherein:
Z4 is N or CR5;
R5 is halo, -CN, -R,-OR, -S(0)nR, -COOR, -CONR2, -NR2, optionally substituted aryl, or optionally substituted heteroaryl;
R2, R3 and R4 are each independently selected from H and optionally substituted CI -CI O alkyl;
X is O, S, or NR6;
Y is O or S or NR10;
Z is O or S;
R6 and R10 are each independently selected from H, CN, optionally substituted C 1 -C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C1 -C4 alkoxy, optionally substituted C6-C10 aryl, and optionally substituted heteroaryl;
cCy is a carbocyclic ring which is optionally further substituted;
L4 is -S(0)-NR-, -S(0)2-NR-, -S(0)-0-, -S(0)2-0-, -C(0)-NR-, -C(0)-O, -NR-S(O)-, - NR-S(0)2-, -O-S(O)-, -0-S(0)2-, -NR-C(O)-, or -O-C(O)-;
Ar is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, or optionally substituted heterocyclyl;
R7 is H, or optionally substituted C1 -C4 alkyl;
R1 is selected from H, optionally substituted C 1 -C10 alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; and
each R is independently selected from H and optionally substituted C1 -C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member; and
n is 0, 1 , or 2;
In one embodiment of Formula (V), Z4 is N; R2, R3, and R4 are H; Z and Y are O; X is NH; R1 is cyclopropyl; L4 is -S(0)-NR-, -S(0)2-NR-, -S(0)-0-, -S(0)2-0-, -C(0)-NR-, -C(O)- 0-; Ar is optionally substituted aryl or optionally substituted heteroaryl; R7 is H; and cCy is a cyclohexyl ring.
In one embodiment of Formula (V), Ar-L4- is Ar-S(0)2-NR-.
In any one of the preceding embodiments of Formula (V), Ar is optionally substituted phenyl.
In one embodim ctural Formula (VI):
Figure imgf000040_0001
(VI), wherein:
Z4 is N or CR5;
R5 is halo, -CN, -R, -OR, -S(0)nR, -C(0)OR, -C(0)NR2, -NR2 or -Ar;
each R is independently selected from H and optionally substituted C1-C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member, n is 0, 1 , or 2;
R2, R3 and R4 are each independently selected from H and optionally substituted CI -CI O alkyl;
X is O, S, or NR6;
Y is O, S, or NR10;
R6 and R10 are each independently selected from H, CN, optionally substituted C 1 -C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C 1 -C4 alkoxy, optionally substituted C6-C 10 aryl, and optionally substituted heteroaryl;
Z is O or S;
Ar is an optionally substituted aryl; and
R1B is selected from H, optionally substituted C1 -C10 alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, or optionally substituted heteroarylalkyl.
In one emb ral Formula (Via):
Figure imgf000041_0001
(Via), wherein
each A is independently optionally substituted C1 -C 10 alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted C2-C10 alkenyl, optionally substituted C2-C10 alkynyl, -NR2; -OR, -S(0)nR, -NR-S(0)nR, halo, -CN, -N02, - C(0)OR, -0-C(0)-R, -C(0)NR2, -NR-C(0)R, or -C(0)R;
c is 0, 1 , 2, 3, 4, or 5;
each n is independently 0, 1 , or 2; and
each R is independently H or optionally substituted C1-C4 alkyl; or alternatively, the two
R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member.
In one embodiment of Formula (VI) or (Via), R3 is -alkylene-0-C(0)-R3a, wherein R3a is optionally substituted C1 -C4 alkyl, optionally substituted C1-C4 heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, or optionally substituted heterocyclyl.
In one embodiment of Formula (Via), -C(0)-R3a is a carboxyl terminus of an amino acid. One embodiment of the amino acid is L- Alanine.
In certain specific embodiments, the present compound is selected from the species disclosed in the specification.
In one embodiment of the present invention, the compounds of Formula (I) or any of its subgenera do not include any of the-compound species, i.e., specific compounds, described in U.S. Patent Application No. 12/946,759. In another embodiment, the compounds of Formula (I) do not include any of the compounds listed in Table X below:
Figure imgf000042_0001
Figure imgf000043_0001
Figure imgf000044_0001
Figure imgf000045_0001
Figure imgf000046_0001
Figure imgf000047_0001
Figure imgf000048_0001
Figure imgf000049_0001
Figure imgf000050_0001
Figure imgf000051_0001
Figure imgf000052_0001
Figure imgf000053_0001
52
Figure imgf000054_0001
Figure imgf000055_0001
Figure imgf000056_0001
Figure imgf000057_0001
Figure imgf000058_0001
Figure imgf000059_0001
Figure imgf000060_0001
Figure imgf000061_0001
Figure imgf000062_0001
Figure imgf000063_0001
Figure imgf000064_0001
63
Figure imgf000065_0001
TABLE X
Figure imgf000066_0001
Figure imgf000067_0001
Figure imgf000068_0001
Figure imgf000069_0001
Figure imgf000070_0001
TABLE X
Figure imgf000071_0001
Figure imgf000072_0001
Figure imgf000073_0001
Figure imgf000074_0001
Figure imgf000075_0001
Figure imgf000076_0001
TABLE X
Figure imgf000077_0001
Figure imgf000078_0001
Figure imgf000079_0001
Figure imgf000080_0001
Figure imgf000081_0001
Figure imgf000082_0001
Figure imgf000083_0001
TABLE X
Figure imgf000084_0001
Figure imgf000085_0001
Figure imgf000086_0001
Figure imgf000087_0001
Figure imgf000088_0001
Figure imgf000089_0001
Figure imgf000090_0001
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Utilities of the Compounds:
In another aspect, the invention provides a pharmaceutical composition comprising any of the above-described compounds, admixed with a pharmaceutically acceptable excipient. In another aspect, the invention provides a method to treat cancer, a vascular disorder, inflammation, infection, pain, or an immunological disorder comprising administering to a subject in need of such treatment, an effective amount of any of the above-described compounds.
The compounds of the invention are useful as medicaments, and are useful for the manufacture of medicaments, including medicaments to treat conditions disclosed herein, such as cancers, inflammatory conditions, infections, pain, and immunological disorders.
The terms "treat" and "treating" as used herein refer to ameliorating, alleviating, lessening, and removing symptoms of a disease or condition. A candidate molecule or compound described herein may be in a therapeutically effective amount in a formulation or medicament, which is an amount that can lead to a biological effect, such as apoptosis of certain cells (e.g., cancer cells), reduction of proliferation of certain cells, or lead to ameliorating, alleviating, lessening, or removing symptoms of a disease or condition, for example. The terms also can refer to reducing or stopping a cell proliferation rate (e.g., slowing or halting tumor growth) or reducing the number of proliferating cancer cells (e.g. , removing part or all of a tumor).
These terms also are applicable to reducing a titre of a microorganism in a system (i.e., cell, tissue, or subject) infected with a microorganism, reducing the rate of microbial propagation, reducing the number of symptoms or an effect of a symptom associated with the microbial infection, and/or removing detectable amounts of the microbe from the system.
Examples of microorganisms include but are not limited to virus, bacterium and fungus.
The compounds of the invention have activities to modulate protein kinases, in particular CK2 activity and/or Pim activity. In some embodiments, the compounds of the invention specifically inhibit the activity of C 2, but not Pim, e.g., more than 100, 90, 80, 70, 60, 50, 40, 30, 20, or 10 fold difference between C 2 inhibition vs. Pim inhibition. In some embodiments, the compounds of the invention specifically inhibit the acitivity of Pim, but not CK.2, e.g., more than 100, 90, 80, 70, 60, 50, 40, 30, 20, or 10 fold difference between Pim inhibition vs. C 2 inhibition. In some embodiments, the compounds of the invention inhibit the activity of C 2 as well as Pim.
The compounds of the invention can be used to modulate the activity of C 2 and/or Pim, e.g. , inhibit the activity of C 2 and/or Pim in a cell, e.g. , in vivo or in vitro. In some embodiments, compounds of the invention can be used to modulate the activity of C 2, e.g., inhibit the activity of CK2 without substantially interfering or changing the activity of Pim. In some embodiments, compounds of the invention can be used to modulate the activity of Pim, e.g., inhibit the activity of Pim without substantially interfering or changing the activity of C 2. In some embodiments, compounds of the invention can be used to modulate the activity of CK2 and Pim, e.g., inhibit the activity of C 2 and Pim.
The compounds of the invention are thus useful to treat infections by certain pathogens, including protozoans and viruses. The invention thus provides methods for treating protozoal disorders such as protozoan parasitosis, including infection by parasitic protozoa responsible for neurological disorders such as schizophrenia, paranoia, and encephalitis in immunocompromised patients, as well as Chagas' disease. It also provides methods to treat various viral diseases, including human immunodeficiency virus type 1 (HIV-1 ), human papilloma viruses (HPVs), herpes simplex virus (HSV), Epstein-Barr virus (EBV), human cytomegalovirus, hepatitis C and B viruses, influenza virus, Borna disease virus, adenovirus, coxsackievirus, coronavirus and varicella zoster virus. The methods for treating these disorders comprise administering to a subject in need thereof an effective amount of a compound of the present invention.
As used herein, the term "apoptosis" refers to an intrinsic cell self-destruction or suicide program. In response to a triggering stimulus, cells undergo a cascade of events including cell shrinkage, blebbing of cell membranes and chromatic condensation and fragmentation. These events culminate in cell conversion to clusters of membrane-bound particles (apoptotic bodies), which are thereafter engulfed by macrophages.
The invention in part provides pharmaceutical compositions comprising at least one compound within the scope of the invention as described herein, and methods of using compounds described herein.
In addition, the invention in part provides methods for identifying a candidate molecule that interacts with a CK2, which comprises contacting a composition containing a CK.2 protein and a molecule described herein with a candidate molecule and determining whether the amount of the molecule described herein that interacts with the protein is modulated, whereby a candidate molecule that modulates the amount of the molecule described herein that interacts with the protein is identified as a candidate molecule that interacts with the protein.
Also provided by the invention are methods for modulating certain protein kinase activities. Protein kinases catalyze the transfer of a gamma nhosphate from adenosine triphosphate to a serine or threonine amino acid (serine/threonine protein kinase), tyrosine amino acid (tyrosine protein kinase), tyrosine, serine or threonine (dual specificity protein kinase) or histidine amino acid (histidine protein kinase) in a peptide or protein substrate. Thus, included herein are methods which comprise contacting a system comprising a protein kinase protein with a compound described herein in an amount effective for modulating (e.g., inhibiting) the activity of the protein kinase. In some embodiments, the activity of the protein kinase is the catalytic activity of the protein (e.g., catalyzing the transfer of a gamma phosphate from adenosine triphosphate to a peptide or protein substrate). In certain embodiments, provided are methods for identifying a candidate molecule that interacts with a protein kinase, which comprise:
contacting a composition containing a protein kinase and a compound described herein with a candidate molecule under conditions in which the compound and the protein kinase interact, and determining whether the amount of the compound that interacts with the protein kinase is modulated relative to a control interaction between the compound and the protein kinase without the candidate molecule, whereby a candidate molecule that modulates the amount of the compound interacting with the protein kinase relative to the control interaction is identified as a candidate molecule that interacts with the protein kinase. Systems in such embodiments can be a cell-free system or a system comprising cells (e.g., in vitro). The protein kinase, the compound or the molecule in some embodiments is in association with a solid phase. In certain embodiments, the interaction between the compound and the protein kinase is detected via a detectable label, where in some embodiments the protein kinase comprises a detectable label and in certain embodiments the compound comprises a detectable label. The interaction between the compound and the protein kinase sometimes is detected without a detectable label.
Provided also are compositions of matter comprising a protein kinase and a compound described herein. In some embodiments, the protein kinase in the composition is a serine- threonine protein kinase. In some embodiments, the protein kinase in the composition is, or contains a subunit (e.g. , catalytic subunit, SH2 domain, SH3 domain) of, C 2. In certain embodiments the composition is cell free and sometimes the protein kinase is a recombinant protein.
The protein kinase can be from any source, such as cells from a mammal, ape or human, for example. Examples of serine-threonine protein kinases that can be inhibited, or may potentially be inhibited, by compounds disclosed herein '"f 'ude without limitation human versions of C 2, or CK2a2. A serine-threonine protein kinase sometimes is a member of a subfamily containing one or more of the following amino acids at positions corresponding to those listed in human CK2: leucine at position 45, methionine at position 163 and isoleucine at position 174. Nucleotide and amino acid sequences for protein kinases and reagents are publicly available (e.g., World Wide Web URLs ncbi.nlm.nih.gov/sites/entrez and Invitrogen.com, each last visited December 2, 2009).
The invention also in part provides methods for treating a condition related to aberrant cell proliferation. For example, provided are methods of treating a cell proliferative condition in a subject, which comprises administering a compound described herein to a subject in need thereof in an amount effective to treat the cell proliferative condition. The subject may be a research animal (e.g., rodent, dog, cat, monkey), optionally containing a tumor such as a xenograft tumor (e.g., human tumor), for example, or may be a human. A cell proliferative condition sometimes is a tumor, e.g., solid or circulating tumor or non-tumor cancer, including but not limited to, cancers of the colorectum, breast, lung, liver, pancreas, lymph node, colon, prostate, brain, head and neck, skin, liver, kidney, blood, ovary and heart (e.g., leukemia, lymphoma, carcinoma).
Compounds and compositions of the invention may be used alone or in combination with anticancer or other agents, such as a palliative agents, that are typically administered to a patient being treated for cancer, as further described herein.
Also provided are methods for treating a condition related to inflammation or pain. For example, methods are provided for treating pain in a subject, which comprise administering a compound described herein to a subject in need thereof in an amount effective to treat the pain. Provided also are methods of treating inflammation in a subject, which comprise administering a compound described herein to a subject in need thereof in an amount effective to treat the inflammation. The subject may be a research animal (e.g., rodent, dog, cat, monkey), for example, or may be a human. Conditions associated with inflammation and pain include without limitation acid reflux, heartburn, acne, allergies and allergen sensitivities, Alzheimer's disease, asthma, atherosclerosis, bronchitis, carditis, celiac disease, chronic pain, Crohn's disease, cirrhosis, colitis, dementia, dermatitis, diabetes, dry eyes, edema, emphysema, eczema, fibromyalgia, gastroenteritis, gingivitis, heart disease, hepatitis, high blood pressure, insulin resistance, interstitial cystitis, joint pain/arthritis/rheumatr>i arthritis, metabolic syndrome (syndrome X), myositis, nephritis, obesity, osteopenia, glomerulonephritis (GN), juvenile cystic kidney disease, and type 1 nephronophthisis (NPHP), osteoporosis, Parkinson's disease, Guam- Parkinson dementia, supranuclear palsy, Kuf s disease, and Pick's disease, as well as memory impairment, brain ischemia, and schizophrenia, periodontal disease, polyarteritis, polychondritis, psoriasis, scleroderma, sinusitis, Sjogren's syndrome, spastic colon, systemic candidiasis, tendonitis, urinary track infections, vaginitis, inflammatory cancer (e.g., inflammatory breast cancer) and the like.
Methods for determining and monitoring effects of compounds herein on pain or inflammation are known. For example, formalin-stimulated pain behaviors in research animals can be monitored after administration of a compound described herein to assess treatment of pain (e.g. , Li et al., Pain 115(1-2): 182-90 (2005)). Also, modulation of pro-inflammatory molecules {e.g., IL-8, GRO-alpha, MCP- 1 , TNFalpha and iNOS) can be monitored after administration of a compound described herein to assess treatment of inflammation (e.g., Parhar et al., Int J Colorectal Dis. 22(6): 601 -9 (2006)), for example. Thus, also provided are methods for determining whether a compound herein reduces inflammation or pain, which comprise contacting a system with a compound described herein in an amount effective for modulating (e.g., inhibiting) the activity of a pain signal or inflammation signal.
Provided also are methods for identifying a compound that reduces inflammation or pain, which comprise: contacting a system with a compound as described herein; and detecting a pain signal or inflammation signal, whereby a compound that modulates the pain signal relative to a control molecule is identified as a compound that reduces inflammation of pain. Non-limiting examples of pain signals are formalin-stimulated pain behaviors and examples of inflammation signals include without limitation a level of a pro-inflammatory molecule. The invention thus in part pertains to methods for modulating angiogenesis in a subject, and methods for treating a condition associated with aberrant angiogenesis in a subject.
CK2 has also been shown to play a role in the pathogenesis of atherosclerosis, and may prevent atherogenesis by maintaining laminar shear stress flow. C 2 plays a role in
vascularization, and has been shown to mediate the hypoxia-induced activation of histone deacetylases (HDACs). CK2 is also involved in diseases relating to skeletal muscle and bone tissue, including, e.g. , cardiomyocyte hypertrophy, heart failure, impaired insulin signaling and insulin resistance, hypophosphatemia and inadequate bon^ matrix mineralization. Thus in one aspect, the invention provides methods to treat each of these conditions, comprising administering to a subject in need of such treatment an effect amount of a CK2 inhibitor, such as a compound as described herein, such as a compound of Formula (I), (II), (Ila), (lib), (lie), (lid), (III), (Ilia), (Illb), (IV), (V), (VI), or (Via) .
The invention also in part pertains to methods for modulating an immune response in a subject, and methods for treating a condition associated with an aberrant immune response in a subject. Thus, provided are methods for determining whether a compound herein modulates an immune response, which comprise contacting a system with a compound described herein in an amount effective for modulating (e.g., inhibiting) an immune response or a signal associated with an immune response. Signals associated with immunomodulatory activity include, e.g., stimulation of T-cell proliferation, suppression or induction of cytokines, including, e.g., interleukins, interferon-γ and TNF. Methods of assessing immunomodulatory activity are known in the art.
Also provided are methods for treating a condition associated with an aberrant immune response in a subject, which comprise administering a compound described herein to a subject in need thereof in an amount effective to treat the condition. Conditions characterized by an aberrant immune response include without limitation, organ transplant rejection, asthma, autoimmune disorders, including rheumatoid arthritis, multiple sclerosis, myasthenia gravis, systemic lupus erythematosus, scleroderma, polymyositis, mixed connective tissue disease (MCTD), Crohn's disease, and ulcerative colitis. In certain embodiments, an immune response may be modulated by administering a compound herein in combination with a molecule that modulates (e.g., inhibits) the biological activity of an mTOR pathway member or member of a related pathway (e.g., mTOR, PI3 kinase, AKT). In certain embodiments the molecule that modulates the biological activity of an mTOR pathway member or member of a related pathway is rapamycin. In certain embodiments, provided herein is a composition comprising a compound described herein in combination with a molecule that modulates the biological activity of an mTOR pathway member or member of a related pathway, such as rapamycin, for example.
Compositions and Routes of Administration
In another aspect, the invention provides pharmaceutical compositions (i.e.,
formulations). The pharmaceutical compositions can comprise a compound of any of Formula (I), (II), (Ha), (lib), (lie), (lid), (III), (Ilia), (Illb), (IV), (V), (VI), and (Via), as described herein which is admixed with at least one pharmaceutically acceptable excipient or carrier. Frequently, the composition comprises at least two pharmaceutically acceptable excipients or carriers.
While the compositions and methods of the present invention will typically be used in therapy for human patients, they may also be used in veterinary medicine to treat similar or identical diseases. The compositions may, for example, be used to treat mammals, including, but not limited to, primates and domesticated mammals. The compositions may, for example be used to treat herbivores. The compositions of the present invention include geometric and optical isomers of one or more of the drugs, wherein each drug is a racemic mixture of isomers or one or more purified isomers.
Pharmaceutical compositions suitable for use in the present invention include
compositions wherein the active ingredients are contained in an effective amount to achieve the intended purpose. Determination of the effective amounts is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein.
The compounds of the present invention may exist as pharmaceutically acceptable salts.
The present invention includes such salts. The term "pharmaceutically acceptable salts" is meant to include salts of active compounds which are prepared with relatively nontoxic acids or bases, depending on the particular substituent moieties found on the compounds described herein. When compounds of the present invention contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Included are base addition salts such as sodium, potassium, calcium, ammonium, organic amino, or magnesium salt, or a similar salt. When compounds of the present invention contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids, for example, acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p- tolylsulfonic, citric, tartaric, methanesulfonic, and the likp A\so included are salts of amino acids such as arginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like (see, for example, Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977, 66, 1 -19). Certain specific compounds of the present invention contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.
Examples of applicable salt forms include hydrochlorides, hydrobromides, sulfates, methanesulfonates, nitrates, maleates, acetates, citrates, fumarates, tartrates (eg (+)-tartrates, (-)- tartrates or mixtures thereof, including racemic mixtures), succinates, benzoates and salts with amino acids such as glutamic acid. These salts may be prepared by methods known to those skilled in art.
The neutral forms of the compounds are preferably regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents.
The pharmaceutically acceptable esters in the present invention refer to non-toxic esters, preferably the alkyl esters such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl or pentyl esters, of which the methyl ester is preferred. However, other esters such as phenyl-Ci-5 alkyl may be employed if desired. Ester derivatives of certain compounds may act as prodrugs which, when absorbed into the bloodstream of a warm-blooded animal, may cleave in such a manner as to release the drug form and permit the drug to afford improved therapeutic efficacy.
Certain compounds of the present invention can exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to unsolvated forms and are encompassed within the scope of the present invention. Certain compounds of the present invention may exist in multiple crystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated by the present invention and are intended to be within the scope of the present invention.
When used as a therapeutic the compounds described herein often are administered with a physiologically acceptable carrier. A physiologically acceptable carrier is a formulation to which the compound can be added to dissolve it or otherwise facilitate its administration.
Examples of physiologically acceptable carriers include, but are not limited to, water, saline, and physiologically buffered saline. In addition to salt forms, the present invention provides compounds that are in a prodrug form. Prodrugs of the compounds described herein are those compounds that readily undergo chemical changes under physiological conditions to provide the compounds of the present invention. Additionally, prodrugs can be converted to the compounds of the present invention by chemical or biochemical methods in an ex vivo environment. For example, prodrugs can be slowly converted to the compounds of the present invention when placed in a transdermal patch reservoir with a suitable enzyme or chemical reagent.
A compound of the present invention can be formulated as a pharmaceutical composition. Such a pharmaceutical composition can then be administered orally, parenterally, by inhalation spray, rectally, or topically in dosage unit formulations containing conventional nontoxic pharmaceutically acceptable carriers, adjuvants, and vehicles as desired.. The amount of active ingredient that can be combined with the carrier materials to produce a single dosage form varies depending upon the mammalian host treated and the particular mode of administration. Topical administration can also involve the use of transdermal administration such, as transdermal patches or iontophoresis devices. The term parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrasternal injection, or infusion techniques. Formulation of drugs is discussed in, for example, Hoover, John E., REMINGTON'S PHARMACEUTICAL
SCIENCES, Mack Publishing Co., Easton, Pa.; 1975. Other examples of drug formulations can be found in Liberman, H. A. and Lachman, L., Eds., PHARMACEUTICAL DOSAGE FORMS, Marcel Decker, New York, N.Y., 1980.
Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions can be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation can also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1 ,3-butanediol. Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables. Dimethyl acetamide, surfactants including ionic and non-ionic detergents, polyethylene glycols can be used. Mixtures of solvents and wetting agents such as those discussed above are also useful . Suppositories for rectal administration of the drug can be prepared by mixing the drug with a suitable nonirritating excipient such as cocoa butter, synthetic mono- di- or triglycerides, fatty acids and polyethylene glycols that are sold at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum and release the drug.
Solid dosage forms for oral administration can include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the compounds of this invention are ordinarily combined with one or more adjuvants appropriate to the indicated route of administration. If administered per os, a compound of the invention can be admixed with lactose, sucrose, starch powder, cellulose esters of alkanoic acids, cellulose alkyl esters, talc, stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric and sulfuric acids, gelatin, acacia gum, sodium alginate, polyvinylpyrrolidone, and/or polyvinyl alcohol, and then tableted or encapsulated for convenient administration. Such capsules or tablets can contain a controlled- release formulation as can be provided in a dispersion of active compound in
hydroxypropylmethyl cellulose. In the case of capsules, tablets, and pills, the dosage forms can also comprise buffering agents such as sodium citrate, magnesium or calcium carbonate or bicarbonate. Tablets and pills can additionally be prepared with enteric coatings.
For therapeutic purposes, formulations for parenteral administration can be in the form of aqueous or non-aqueous isotonic sterile injection solutions or suspensions. These solutions and suspensions can be prepared from sterile powders or granules having one or more of the carriers or diluents mentioned for use in the formulations for oral administration. A compound of the invention can be dissolved in water, polyethylene glycol, propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride, and/or various buffers. Other adjuvants and modes of administration are well and widely known in the pharmaceutical art.
Liquid dosage forms for oral administration can include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art, such as water. Such compositions can also comprise adjuvants, such as wetting agents, emulsifying and suspending agents, and sweetening, flavoring, and perfuming agents.
The dosage regimen utilizing the compounds of the present invention in combination with an anticancer agent is selected in accordance with a variety of factors including type, species, age, weight, sex and medical condition of the pa†ipnt; the severity of the condition to be treated; the route of administration; the renal and hepatic function of the patient; and the particular compound or salt or ester thereof employed. A consideration of these factors is well within the purview of the ordinarily skilled clinician for the purpose of determining the therapeutically effective dosage amounts to be given to a person in need of the instant combination therapy.
Various sustained release systems for drugs have also been devised, and can be applied to compounds of the invention. See, for example, U.S. Patent No. 5,624,677, the methods of which are incorporated herein by reference.
Systemic administration may also include relatively noninvasive methods such as the use of suppositories, transdermal patches, transmucosal delivery and intranasal administration. Oral administration is also suitable for compounds of the invention. Suitable forms include syrups, capsules, tablets, as is understood in the art.
For administration to animal or human subjects, the appropriate dosage of a compound described above often is 0.01 -15 mg/kg, and sometimes 0.1 -10 mg/kg. In some embodiments, a suitable dosage of the compound of the invention for an adult patient will be between 1 and 1000 mg per dose, frequently between 10 and 300 mg, and the dosage may be administered 1 -4 times per day. Dosage levels are dependent on the nature of the condition, drug efficacy, the condition of the patient, the judgment of the practitioner, and the frequency and mode of administration; optimization of such parameters is within the ordinary level of skill in the art.
Therapeutic Combinations
Compounds of the invention may be used alone or in combination with another therapeutic agent. The invention provides methods to treat conditions such as cancer, inflammation and immune disorders by administering to a subject in need of such treatment a therapeutically effective amount of a therapeutic agent useful for treating said disorder and administering to the same subject a therapeutically effective amount of a modulator of the present invention, i.e., a compound of the invention. The therapeutic agent and the modulator may be "co-administered", i.e, administered together, either as separate pharmaceutical compositions or admixed in a single pharmaceutical composition. By "administered together", the therapeutic agent and the modulator may also be administered separately, including at different times and with different frequencies. The modulator may be administered by any known route, such as orally, intravenously, intramuscularly1, nasally, and the like; and the therapeutic agent may also be administered by any conventional route. In many embodiments, at least one and optionally both of the modulator and the therapeutic agent may be administered orally. Preferably, the modulator is an inhibitor, and it may inhibit either one of C 2 and Pirn, or both of them to provide the treatment effects described herein.
In certain embodiments, a "modulator" as described above may be used in combination with a therapeutic agent that can act by binding to regions of DNA that can form certain quadruplex structures. In such embodiments, the therapeutic agents have anticancer activity on their own, but their activity is enhanced when they are used in combination with a modulator. This synergistic effect allows the therapeutic agent to be administered in a lower dosage while achieving equivalent or higher levels of at least one desired effect.
A modulator may be separately active for treating a cancer. For combination therapies described above, when used in combination with a therapeutic agent, the dosage of a modulator will frequently be two-fold to ten-fold lower than the dosage required when the modulator is used alone to treat the same condition or subject. Determination of a suitable amount of the modulator for use in combination with a therapeutic agent is readily determined by methods known in the art.
Compounds and compositions of the invention may be used in combination with anticancer or other agents, such as palliative agents, that are typically administered to a patient being treated for cancer. Such "anticancer agents" include, e.g. , classic chemotherapeutic agents, as well as molecular targeted therapeutic agents, biologic therapy agents, and radiotherapeutic agents.
When a compound or composition of the invention is used in combination with an anticancer agent to another agent, the present invention provides, for example, simultaneous, staggered, or alternating treatment. Thus, the compound of the invention may be administered at the same time as an anticancer agent, in the same pharmaceutical composition; the compound of the invention may be administered at the same time as the anticancer agent, in separate pharmaceutical compositions; the compound of the invention may be administered before the anticancer agent, or the anticancer agent may be administered before the compound of the invention, for example, with a time difference of seconds, minutes, hours, days, or weeks. In examples of a staggered treatment, a course of therapy with the compound of the invention may be administered, followed by a course of therapy with the anticancer agent, or the reverse order of treatment may be used, and more than one series of treatments with each component may also be used. In certain examples of the present invention, one component, for example, the compound of the invention or the anticancer agent, is administered to a mammal while the other component, or its derivative products, remains in the bloodstream of the mammal. For example, the present compound may be administered while the anticancer agent or its derivative products remains in the bloodstream, or the anticancer agent may be administered while the present compound or its derivatives remains in the bloodstream. In other examples, the second component is administered after all, or most of the first component, or its derivatives, have left the bloodstream of the mammal.
The compound of the invention and the anticancer agent may be administered in the same dosage form, e.g., both administered as intravenous solutions, or they may be administered in different dosage forms, e.g. , one compound may be administered topically and the other orally. A person of ordinary skill in the art would be able to discern which combinations of agents would be useful based on the particular characteristics of the drugs and the cancer involved.
Anticancer agents useful in combination with the compounds of the present invention may include agents selected from any of the classes known to those of ordinary skill in the art, including, but not limited to, antimicrotubule agents such as diterpenoids and vinca alkaloids; platinum coordination complexes; alkylating agents such as nitrogen mustards,
oxazaphosphorines, alkylsulfonates, nitrosoureas, and triazenes; antibiotic agents such as anthracyclins, actinomycins and bleomycins; topoisomerase II inhibitors such as
epipodophyllotoxins; antimetabolites such as purine and pyrimidine analogues and anti-folate compounds; topoisomerase I inhibitors such as camptothecins; hormones and hormonal analogues; signal transduction pathway inhibitors; nonreceptor tyrosine kinase angiogenesis inhibitors; immunotherapeutic agents; pro-apoptotic agents; and cell cycle signaling inhibitors; and other agents described below.
Anti-microtubule or anti-mitotic agents are phase specific agents that are typically active against the microtubules of tumor cells during M or the mitosis phase of the cell cycle.
Examples of anti-microtubule agents include, but are not limited to, diterpenoids and vinca alkaloids. Plant alkaloid and terpenoid derived agents include mitotic inhibitors such as the vinca alkaloids vinblastine, vincristine, vindesine, and vinorelbine; and microtubule polymer stabilizers such as the taxanes, including, but not limited to paclitaxel, docetaxel, larotaxel, ortataxel, and tesetaxel.
Diterpenoids, which are derived from natural sources, are phase specific anti - cancer agents that are believed to operate at the G2/ phases of the cell cycle. It is believed that the diterpenoids stabilize the p-tubulin subunit of the microtubules, by binding with this protein. Disassembly of the protein appears then to be inhibited with mitosis being arrested and cell death following.
Examples, of diterpenoids include, but are not limited to, taxanes such as paclitaxel, docetaxel, larotaxel, ortataxel, and tesetaxel. Paclitaxel is a natural diterpene product isolated from the Pacific yew tree Taxus brevifolia and is commercially available as an injectable solution TAXOL®. Docetaxel is a semisynthetic derivative of paclitaxel q. v. , prepared using a natural precursor, 10-deacetyl-baccatin III, extracted from the needle of the European Yew tree.
Docetaxel is commercially available as an injectable solution as TAXOTERE®.
Vinca alkaloids are phase specific anti-neoplastic agents derived from the periwinkle plant. Vinca alkaloids that are believed to act at the M phase (mitosis) of the cell cycle by binding specifically to tubulin. Consequently, the bound tubulin molecule is unable to polymerize into microtubules. Mitosis is believed to be arrested in metaphase with cell death following. Examples of vinca alkaloids include, but are not limited to, vinblastine, vincristine, vindesine, and vinorelbine. Vinblastine, vincaleukoblastine sulfate, is commercially available as VELBAN® as an injectable solution. Vincristine, vincaleukoblastine 22-oxo-sulfate, is commercially available as ONCOVIN® as an injectable solution. Vinorelbine, is commercially available as an injectable solution of vinorelbine tartrate (NAVELBINE®), and is a
semisynthetic vinca alkaloid derivative.
Platinum coordination complexes are non-phase specific anti-cancer agents, which are interactive with DNA. The platinum complexes are believed to enter tumor cells, undergo, aquation and form intra- and interstrand crosslinks with DNA causing adverse biological effects to the tumor. Platinum-based coordination complexes include, but are not limited to cisplatin, carboplatin, nedaplatin, oxaliplatin, satraplatin, and (SP-4-3)-(cis)- amminedichloro-[2- methylpyridine] platinum(II). Cisplatin, cis-diamminedic > oplatinum, is commercially available as PLATINOL® as an injectable solution. Carboplatin, platinum, diammine [1 , 1 - cyclobutane-dicarboxylate(2-)-0,0'], is commercially available as PARAPLATIN® as an injectable solution.
Alkylating agents are generally non-phase specific agents and typically are strong electrophiles. Typically, alkylating agents form covalent linkages, by alkylation, to DNA through nucleophilic moieties of the DNA molecule such as phosphate, amino, sulfhydryl, hydroxyl, carboxyl, and imidazole groups. Such alkylation disrupts nucleic acid function leading to cell death. Examples of alkylating agents include, but are not limited to, alkyl sulfonates such as busulfan; ethyleneimine and methylmelamine derivatives such as altretamine and thiotepa; nitrogen mustards such as chlorambucil, cyclophosphamide, estramustine, ifosfamide, mechlorethamine, melphalan, and uramustine; nitrosoureas such as carmustine, lomustine, and streptozocin; triazenes and imidazotetrazines such as dacarbazine, procarbazine, temozolamide, and temozolomide. Cyclophosphamide, 2-[bis(2-chloroethyl)-amino]tetrahydro-2H- 1 ,3,2- oxazaphosphorine 2-oxide monohydrate, is commercially available as an injectable solution or tablets as CYTOXAN®. Melphalan, 4-[bis(2-chloroethyl)amino]-L-phenylalanine, is commercially available as an injectable solution or tablets as AL ERAN®. Chlorambucil, 4- [bis(2-chloroethyl)amino]-benzenebutanoic acid, is commercially available as LEUKERAN® tablets. Busulfan, 1 ,4-butanediol dimethanesulfonate, is commercially available as
MYLERAN® TABLETS. Carmustine, l ,3-[bis(2-chloroethyl)-l -nitrosourea, is commercially available as single vials of lyophilized material as BiCNU®, 5-(3,3-dimethyl- l -triazeno)- imidazole-4-carboxamide, is commercially available as single vials of material as DTIC-Dome®. Furthermore, alkylating agents include (a) alkylating-like platinum-based chemotherapeutic agents such as cisplatin, carboplatin, nedaplatin, oxaliplatin, satraplatin, and (SP-4-3)-(cis)- amminedichloro-[2-methylpyridine] platinum(II); (b) alkyl sulfonates such as busulfan;
(c) ethyleneimine and methylmelamine derivatives such as altretamine and thiotepa; (d) nitrogen mustards such as chlorambucil, cyclophosphamide, estramustine, ifosfamide, mechlorethamine, trofosamide, prednimustine, melphalan, and uramustine; (e) nitrosoureas such as carmustine, lomustine, fotemustine, nimustine, ranimustine and streptozocin; (f) triazenes and
imidazotetrazines such as dacarbazine, procarbazine, temozolamide, and temozolomide.
Anti-tumor antibiotics are non-phase specific agents which are believed to bind or intercalate with DNA. This may result in stable DNA complexes or strand breakage, which disrupts ordinary function of the nucleic acids, leading to cell death. Examples of anti-tumor antibiotic agents include, but are not limited to, anthracyclines such as daunorubicin (including liposomal daunorubicin), doxorubicin (including liposomal doxorubicin), epirubicin, idarubicin, and valrubicin; streptomyces-related agents such as bleomycin, actinomycin, mithramycin, mitomycin, porfiromycin; and mitoxantrone. Dactinomycin, also know as Actinomycin D, is commercially available in injectable form as COSMEGEN®. Daunorubicin, (8S-cis-)-8-acetyl- 10-[(3-amino-2,3,6-trideoxy-a-L-lyxohexopyranosy l)oxy]-7,8,9, 10-tetrahydro-6,8, 1 1 - trihydroxy- l-methoxy-5, 12-naphthacenedione hydrochloride, is commercially available as a liposomal injectable form as DAU OXOME® or as an injectable as CERUB1DINE®.
Doxorubicin, (8S, 10S)- 10-[(3-amino-2,3,6-trideoxy-a-L-lyxohexopyranosyl)oxy]-8-glycoloyl, 7,8,9, 1 0-tetrahydro-6,8, 1 l -trihydroxy- l -methoxy-5, 12-naphthacenedione hydrochloride, is commercially available in an injectable form as RUBEX® or ADRIAMYC1N RDF®.
Bleomycin, a mixture of cytotoxic glycopeptide antibiotics isolated from a strain of Streptomyces verticillus, is commercially available as BLENOXANE®.
Topoisomerase inhibitors include topoisomerase I inhibitors such as camptothecin, topotecan, irinotecan, rubitecan, and belotecan; and topoisomerase II inhibitors such as etoposide, teniposide, and amsacrine.
Topoisomerase II inhibitors also include, but are not limited to, epipodophyllotoxins, which are phase specific anti-neoplastic agents derived from the mandrake plant.
Epipodophyllotoxins typically affect cells in the S and G2 phases of the cell cycle by forming a ternary complex with topoisomerase II and DNA causing DNA strand breaks. The strand breaks accumulate and cell death follows. Examples of epipodophyllotoxins include, but are not limited to, etoposide, teniposide, and amsacrine. Etoposide, 4'-demethyl-epipodophyllotoxin 9[4,6-0- (R)-ethylidene- -D-. glucopyranoside], is commercially available as an injectable solution or capsules as VePESID® and is commonly known as VP- 16. Teniposide, 4'-demethyl- epipodophyllotoxin 9[4,6-0-(R )-thenylidene-P-D-glucopyranoside], is commercially available as an injectable solution as VUMON® and is commonly known as VM-26.
Further examples of topoisomerase I inhibitors include, but are not limited to camptothecin and camptothecin derivatives, topotecan, irinotecan, rubitecan, belotecan and the various optical forms (i.e., (R), (S) or (R,S)) of 7-(4-methylpiperazino-methylene)- 10, 1 1 - ethylenedioxy-camptothecin, as described in U.S. Patent 6,063,923; 6, 100,273; 5,342,947; 5,559,235; and 5,491 ,237. Irinotecan HC1, (4S)-4, 1 l -diethyl-4-hydroxy-9-[(4- piperidinopiperidino)-carbonyloxy]-l H-pyrano[3',4',6,7]indolizino[l ,2-b]quinoline-3, 14(4H, 12H)-dione hydrochloride, is commercially available as the injectable solution CA PTOSA ®. Irinotecan is a derivative of camptothecin which binds, along with its active metabolite 8N-38, to the topoisomerase I-DNA complex. Topotecan HC1, (S)-10-[(dimethylamino)methyl]-4-ethyl- 4,9-dihydroxy-l H-pyrano[3',4',6,7]indolizino[l ,2-b]quinoIine-3, 14-(4H, 12H)-dione monohydrochloride, is commercially available as the injectable solution HYCA TIN®.
Anti-metabolites include (a) purine analogs such as fludarabine, cladribine,
chlorodeoxyadenosine, clofarabine, mercaptopurine, pentostatin, and thioguanine; (b) pyrimidine analogs such as fluorouracil, gemcitabine, capecitabine, cytarabine, azacitidine, edatrexate, floxuridine, and troxacitabine; (c) antifolates, such as methotrexate, pemetrexed, raltitrexed, and trimetrexate. Anti-metabolites also include thymidylate synthase inhibitors, such as fluorouracil, raltitrexed, capecitabine, floxuridine and pemetrexed; and ribonucleotide reductase inhibitors such as claribine, clofarabine and fludarabine. Antimetabolite neoplastic agents are phase specific anti-neoplastic agents that typically act at S phase (DNA synthesis) of the cell cycle by inhibiting DNA synthesis or by inhibiting purine or pyrimidine base synthesis and thereby limiting DNA synthesis. Consequently, S phase does not proceed and cell death follows. Antimetabolites, include purine analogs, such as fludarabine, cladribine, chlorodeoxyadenosine, clofarabine, mercaptopurine, pentostatin, erythrohydroxynonyladenine, fludarabine phosphate and thioguanine; pyrimidine analogs such as fluorouracil, gemcitabine, capecitabine, cytarabine, azacitidine, edatrexate, floxuridine, and troxacitabine; antifolates, such as methotrexate, pemetrexed, raltitrexed, and trimetrexate. Cytarabine, 4-amino- l -p-D-arabinofuranosyl-2 ( 1 H)- pyrimidinone, is commercially available as CYTOSAR-U® and is commonly known as Ara-C. Mercaptopurine, 1 ,7-dihydro-6H-purine-6-thione monohydrate, is commercially available as PURINETHOL®. Thioguanine, 2-amino-l , 7-dihydro-6H-purine-6-thione, is commercially available as TABLOID®. Gemcitabine, 2'-deoxy-2', 2'-difluorocytidine monohydrochloride (p- isomer), is commercially available as GE ZAR®.
Hormonal therapies include (a) androgens such as fluoxymesterone and testolactone; (b) antiandrogens such as bicalutamide, cyproterone, flutamide, and nilutamide; (c) aromatase inhibitors such as aminoglutethimide, anastrozole, exemestane, formestane, and letrozole;
(d) corticosteroids such as dexamethasone and prednisone p) estrogens such as diethylstilbestrol; (f) antiestrogens such as fulvestrant, raloxifene, tamoxifen, and toremifine;
(g) LHRH agonists and antagonists such as buserelin, goserelin, leuprolide, and triptorelin;
(h) progestins such as medroxyprogesterone acetate and megestrol acetate; and (i) thyroid hormones such as levothyroxine and liothyronine. Hormones and hormonal analogues are useful compounds for treating cancers in which there is a relationship between the hormone(s) and growth and/or lack of growth of the cancer. Examples of hormones and hormonal analogues useful in cancer treatment include, but are not limited to, androgens such as fluoxymesterone and testolactone; antiandrogens such as bicalutamide, cyproterone, flutamide, and nilutamide;
aromatase inhibitors such as aminoglutethimide, anastrozole, exemestane, formestane, vorazole, and letrozole; corticosteroids such as dexamethasone, prednisone and prednisolone; estrogens such as diethylstilbestrol; antiestrogens such as fulvestrant, raloxifene, tamoxifen, toremifine, droloxifene, and iodoxyfene, as well as selective estrogen receptor modulators (SERMS) such those described in U.S. Patent Nos. 5,681 ,835, 5,877,219, and 6,207,716; 5a-reductases such as finasteride and dutasteride; gonadotropin-releasing hormone (GnRH) and analogues thereof which stimulate the release of leutinizing hormone (LH) and/or follicle stimulating hormone (FSH), for example LHRH agonists and antagonists such as buserelin, goserelin, leuprolide, and triptorelin; progestins such as medroxyprogesterone acetate and megestrol acetate; and thyroid hormones such as levothyroxine and liothyronine.
Signal transduction pathway inhibitors are those inhibitors, which block or inhibit a chemical process which evokes an intracellular change, such as cell proliferation or
differentiation. Signal trahduction inhibitors useful in the present invention include, e.g., inhibitors of receptor tyrosine kinases, non-receptor tyrosine kinases, SH2/SH3 domain blockers, serine/threonine kinases, phosphotidyl inositol-3 kinases, myo-inositol signaling, and Ras oncogenes.
Molecular targeted agents include (a) receptor tyrosine kinase ('RTK') inhibitors, such as inhibitors of EGFR, including erlotinib, gefitinib, and neratinib; inhibitors of VEGFR including vandetanib, semaxinib, and cediranib; and inhibitors of PDGFR; further included are RTK inhibitors that act at multiple receptor sites such as lapatinib, which inhibits both EGFR and HER2, as well as those inhibitors that act at each of C-kit, PDGFR and VEGFR, including but not limited to axitinib, sunitinib, sorafenib and toceranib; also included are inhibitors of BCR- ABL, c-kit and PDGFR, such as imatinib; (b) FKBP binding agents, such as an immunosuppressive macrolide antibiotic, including bafilomycin, rapamycin (sirolimus) and everolimus; (c) gene therapy agents, antisense therapy agents, and gene expression modulators such as the retinoids and rexinoids, e.g. , adapalene, bexarotene, trans-retinoic acid, 9-cis-retinoic acid, and N-(4-hydroxyphenyl)retinamide; (d) phenotype-directed therapy agents, including monoclonal antibodies such as alemtuzumab, bevacizumab, cetuximab, ibritumomab tiuxetan, rituximab, and trastuzumab; (e) immunotoxins such as gemtuzumab ozogamicin;
(f) radioimmunoconjugates such as 1311-tositumomab; and (g) cancer vaccines.
Several protein tyrosine kinases catalyse the phosphorylation of specific tyrosyl residues in various proteins involved in the regulation of cell growth. Such protein tyrosine kinases can be broadly classified as receptor or non-receptor kinases. Receptor tyrosine kinases are transmembrane proteins having an extracellular ligand binding domain, a transmembrane domain, and a tyrosine kinase domain. Receptor. tyrosine kinases are involved in the regulation of cell growth and are sometimes termed growth factor receptors.
Inappropriate or uncontrolled activation of many of these kinases, for example by over- expression or mutation, has been shown to result in uncontrolled cell growth. Accordingly, the aberrant activity of such kinases has been linked to malignant tissue growth. Consequently, inhibitors of such kinases could provide cancer treatment methods.
Growth factor receptors include, for example, epidermal growth factor receptor (EGFr), platelet derived growth factor receptor (PDGFr), erbB2, erbB4, vascular endothelial growth factor receptor (VEGFr), tyrosine kinase with immunoglobulin-like and epidermal growth factor homology domains (TIE-2), insulin growth factor -I (IGFI) receptor, macrophage colony stimulating factor (cfms), BTK, ckit, cmet, fibroblast growth factor (FGF) receptors, Trk receptors (TrkA, TrkB, and TrkC), ephrin (eph) receptors, and the RET protooncogene.
Several inhibitors of growth receptors are under development and include ligand antagonists, antibodies, tyrosine kinase inhibitors and anti-sense oligonucleotides. Growth factor receptors and agents that inhibit growth factor receptor function are described, for instance, in Kath, John C, Exp. Opin. Ther. Patents (2000) 10(6):803-818; Shawver et al., Drug Discov. Today ( 1997), 2(2):50-63; and Lofts, F. J. et a!., "Growth factor receptors as targets", New Molecular Targets for Cancer Chemotherapy, ed. Workman, Paul and Kerr, David, CRC press 1994, London. Specific examples of receptor tyrosine kinase inhibitors include, but are not limited to, sunitinib, erlotinib, gefitinib, and imatinib. Tyrosine kinases which are not growth factor receptor kinases are termed non-receptor tyrosine kinases. Non-receptor tyrosine kinases useful in the present invention, which are targets or potential targets of anti-cancer drugs, include cSrc, Lck, Fyn, Yes, Jak, cAbl, FAK (Focal adhesion kinase), Brutons tyrosine kinase, and Bcr-Abl. Such non-receptor kinases and agents which inhibit non-receptor tyrosine kinase function are described in Sinh, S. and Corey, S.J., J. Hematotherapy & Stem Cell Res. (1999) 8(5): 465 - 80; and Bolen, J.B., Brugge, J.S., Annual Review of Immunology. (1997) 15: 371-404.
SH2/SH3 domain blockers are agents that disrupt SH2 or SH3 domain binding in a variety of enzymes or adaptor proteins including, PI3-K p85 subunit, Src family kinases, adaptor molecules (She, Crk, Nek, Grb2) and Ras-GAP. SH2/SH3 domains as targets for anti-cancer drugs are discussed in Smithgall, T.E., J Pharmacol. Toxicol. Methods. (1995), 34(3): 125-32. Inhibitors of Serine/Threonine Kinases including MAP kinase cascade blockers which include blockers of Raf kinases (rafk), Mitogen or Extracellular Regulated Kinase (MEKs), and
Extracellular Regulated Kinases (ERKs); and Protein kinase C family member blockers including blockers of PKCs (alpha, beta, gamma, epsilon, mu, lambda, iota, zeta). IkB kinase family (IKKa, IKKb), PKB family kinases, AKT kinase family members, and TGF beta receptor kinases. Such Serine/Threonine kinases and inhibitors thereof are described in Yamamoto, T., Taya, S., Kaibuchi, K., J. Biochemistry. (1999) 126 (5): 799-803; Brodt, P, Samani, A, & Navab, R, Biochem. Pharmacol. (2000) 60: 1 101 -1 107; Massague, J., Weis-Garcia, F., Cancer Surv. (1996) 27:41 -64; Philip, P.A, and Harris, AL, Cancer Treat. Res. (1995) 78: 3-27; Lackey, K. et al. Bioorg. Med. Chem. Letters, (2000) 10(3): 223-226; U.S. Patent No. 6,268,391 ; and
Martinez-Lacaci, I., et al., Int. J. Cancer (2000), 88(1): 44-52. Inhibitors of Phosphotidyl inositol-3 Kinase family members including blockers of PI3- kinase, ATM, DNA-PK, and Ku are also useful in the present invention. Such kinases are discussed in Abraham, RT. Current Opin. Immunol. (1996), 8(3): 412-8; Canman, C.E., Lim, D.S., Oncogene (1998) 17(25): 3301 -8; Jackson, S.P., Int. J. Biochem. Cell Biol. (1997) 29(7):935-8; and Zhong, H. et al., Cancer Res. (2000) 60(6): 1541 -5. Also useful in the present invention are Myo-inositol signaling inhibitors such as phospholipase C blockers and Myoinositol analogues. Such signal inhibitors are described in Powis, G., and Kozikowski A, (1994) New Molecular Targets for Cancer
Chemotherapy, ed., Paul Workman and David Kerr, CRC Press 1994, London. Another group of signal transduction pathway inhibitors are inhibitors of Ras Oncogene. Such inhibitors include inhibitors of farnesyltransferase, geranyl-geranyl transferase, and CAAX proteases as well as anti-sense oligonucleotides, ribozymes and immunotherapy. Such inhibitors have been shown to block ras activation in cells containing wild type mutant ras, thereby acting as antiproliferation agents. Ras oncogene inhibition is discussed in Scharovsky, O.G., Rozados, V.R, Gervasoni, SI, Matar, P., J. Biomed. Sci. (2000) 7(4): 292-8; Ashby, M.N., Curr. Opin. Lipidol. (1998) 9(2): 99 - 102; and Oliff, A., Biochim. Biophys. Acta, (1999) 1423(3):C19-30.
As mentioned above, antibody antagonists to receptor kinase ligand binding may also serve as signal transduction inhibitors. This group of signal transduction pathway inhibitors includes the use of humanized antibodies to the extracellular ligand binding domain of receptor tyrosine kinases. For example Imclone C225 EGFR specific antibody {see Green, M.C. et al., Cancer Treat. Rev., (2000) 26(4): 269-286); Herceptin® erbB2 antibody {see Stern, DF, Breast Cancer Res. (2000) 2(3): 176-183); and 2CB VEGFR2 specific antibody {see Brekken, R.A. et al., Cancer Res. (2000) 60(18):51 17-24).
Non-receptor kinase angiogenesis inhibitors may also find use in the present invention.
Inhibitors of angiogenesis related VEGFR and TIE2 are discussed above in regard to signal transduction inhibitors (both receptors are receptor tyrosine kinases). Angiogenesis in general is linked to erbB2/EGFR signaling since inhibitors of erbB2 and EGFR have been shown to inhibit angiogenesis, primarily VEGF expression. Thus, the combination of an erbB2/EGFR inhibitor with an inhibitor of angiogenesis makes sense. Accordingly, non-receptor tyrosine kinase inhibitors may be used in combination with the EGFR/erbB2 inhibitors of the present invention. For example, anti-VEGF antibodies, which do not recognize VEGFR (the receptor tyrosine kinase), but bind to the ligand; small molecule inhibitors of integrin (alphav beta3) that will inhibit angiogenesis; endostatin and angiostatin (non-RT ) may also prove useful in combination with the disclosed erb family inhibitors. (See Bruns, CJ et al., Cancer Res. (2000), 60(1 1 ): 2926-2935; Schreiber AB, Winkler ME, & Derynck R., Science (1986) 232(4755): 1250- 53; Yen L. et al., Oncogene (2000) 19(31 ): 3460-9).
Agents used in immunotherapeutic regimens may also be useful in combination with the compounds of formula (I). There are a number of immunologic strategies to generate an immune response against erbB2 or EGFR. These strategies are generally in the realm of tumor vaccinations. The efficacy of immunologic approaches r"av be greatly enhanced through combined inhibition of erbB2/EGFR signaling pathways using a small molecule inhibitor. Discussion of the immunologic/tumor vaccine approach against erbB2/EGFR are found in Reilly RT, et a!., Cancer Res. (2000) 60( 13):3569-76; and Chen Y, et al., Cancer Res. (1998)
58(9): 1965-71.
Agents used in pro-apoptotic regimens (e.g., bcl-2 antisense oligonucleotides) may also be used in the combination of the present invention. Members of the Bcl-2 family of proteins block apoptosis. Upregulation of bcl-2 has therefore been linked to chemoresistance. Studies have shown that the epidermal growth factor (EGF) stimulates anti-apoptotic members of the bcl-2 family. Therefore, strategies designed to downregulate the expression of bcl-2 in tumors have demonstrated clinical benefit and are now in Phase II/III trials, namely Genta's G3139 bcl- 2 antisense oligonucleotide. Such pro-apoptotic strategies using the antisense oligonucleotide strategy for bcl-2 are discussed in Waters JS, et al., J. Clin. Oncol. (2000) 18(9): 1812-23; and Kitada S, et al. Antisense Res. Dev. (1994) 4(2): 71 -9.
Cell cycle signalling inhibitors inhibit molecules involved in the control of the cell cycle. A family of protein kinases called cyclin dependent kinases (CDKs) and their interaction with a family of proteins termed cyclins controls progression through the eukaryotic cell cycle. The coordinate activation and inactivation of different cyclin/CDK complexes is necessary for normal progression through the cell cycle. Several inhibitors of cell cycle signalling are under development. For instance, examples of cyclin dependent kinases, including CDK2, CDK4, and CDK6 and inhibitors for the same are described in, for instance, RosaniaGR & Chang Y-T., Exp. Opin. Ther. Patents (2000) 10(2):215-30.
Other molecular targeted agents include FKBP binding agents, such as the
immunosuppressive macrolide antibiotic, rapamycin; gene therapy agents, antisense therapy agents, and gene expression modulators such as the retinoids and rexinoids, e.g. , adapalene, bexarotene, trans-retinoic acid, 9-cisretinoic acid, and N-(4 hydroxyphenyl)retinamide;
phenotype-directed therapy agents, including: monoclonal antibodies such as alemtuzumab, bevacizumab, cetuximab, ibritumomab tiuxetan, rituximab, and trastuzumab; immunotoxins such as gemtuzumab ozogamicin, radioimmunoconjugates such as 131 -tositumomab; and cancer vaccines.
Anti-tumor antibiotics include (a) anthracyclines such as daunorubicin (including liposomal daunorubicin), doxorubicin (including liposorr*1 doxorubicin), epirubicin, idarubicin, and valrubicin; (b) streptomyces-related agents such as bleomycin, actinomycin, mithramycin, mitomycin, porfiromycin; and (c) anthracenediones, such as mitoxantrone and pixantrone. Anthracyclines have three mechanisms of action: intercalating between base pairs of the DNA/RNA strand; inhibiting topoiosomerase II enzyme; and creating iron-mediated free oxygen radicals that damage the DNA and cell membranes. Anthracyclines are generally characterized as topoisomerase II inhibitors.
Monoclonal antibodies include, but are not limited to, murine, chimeric, or partial or fully humanized monoclonal antibodies. Such therapeutic antibodies include, but are not limited to antibodies directed to tumor or cancer antigens either on the cell surface or inside the cell. Such therapeutic antibodies also include, but are not limited to antibodies directed to targets or pathways directly or indirectly associated with C 2. Therapeutic antibodies may further include, but are not limited to antibodies directed to targets or pathways that directly interact with targets or pathways associated with the compounds of the present invention. In one variation, therapeutic antibodies include; but are not limited to anticancer agents such as Abagovomab, Adecatumumab, Afutuzumab, Alacizumab pegol, Alemtuzumab, Altumomab pentetate, Anatumomab mafenatox, Apolizumab, Bavituximab, Belimumab, Bevacizumab, Bivatuzumab mertansine, Blinatumomab, Brentuximab vedotin, Cantuzumab mertansine, Catumaxomab, Cetuximab, Citatuzumab bogatox, Cixutumumab, Clivatuzumab tetraxetan, Conatumumab, Dacetuzumab, Detumomab, Ecromeximab, Edrecolomab, Elotuzumab, Epratuzumab, Ertumaxomab, Etaracizumab, Farletuzumab, Figitumumab, Fresolimumab, Galiximab, Glembatumumab vedotin, Ibritumomab tiuxetan, Intetumumab, Inotuzumab ozogamicin, Ipilimumab, Iratumumab, Labetuzumab, Lexatumumab, Lintuzumab,
Lucatumumab, Lumiliximab, Mapatumumab, Matuzumab, Milatuzumab, Mitumomab, Nacolomab tafenatox, Naptumomab estafenatox, Necitumumab, Nimotuzumab, Ofatumumab, Olaratumab, Oportuzumab monatox, Oregovomab, Panitumumab, Pemtumomab, Pertuzumab, Pintumomab, Pritumumab, Ramucirumab, Rilotumumab, Rituximab, Robatumumab,
Sibrotuzumab, Tacatuzumab tetraxetan, Taplitumomab paptox, Tenatumomab, Ticilimumab, Tigatuzumab, Tositumomab, Trastuzumab, Tremelimumab, Tucotuzumab celmoleukin, Veltuzumab, Volociximab, Votumumab, Zalutumumab, and Zanolimumab. In some embodiments, such therapeutic antibodies include, alemtuzumab, bevacizumab, cetuximab, daclizumab, gemtuzumab, ibritumomab tiuxetan, pantitum, ,rnab, rituximab, tositumomab, and trastuzumab; in other embodiments, such monoclonal antibodies include alemtuzumab, bevacizumab, cetuximab, ibritumomab tiuxetan, rituximab, and trastuzumab; alternately, such antibodies include daclizumab, gemtuzumab, and pantitumumab. In yet another embodiment, therapeutic antibodies useful in the treatment of infections include but are not limited to Afelimomab, Efungumab, Exbivirumab, Felvizumab, Foravirumab, Ibalizumab, Libivirumab, Motavizumab, Nebacumab, Pagibaximab, Palivizumab, Panobacumab, Rafivirumab,
Raxibacumab, Regavirumab, Sevirumab, Tefibazumab, Tuvirumab, and Urtoxazumab. In a further embodiment, therapeutic antibodies can be useful in the treatment of inflammation and/or autoimmune disorders, including, but are not limited to, Adalimumab, Atlizumab,
Atorolimumab, Aselizumab, Bapineuzumab, Basiliximab, Benralizumab, Bertilimumab,
Besilesomab, Briakinumab, Canakinumab, Cedelizumab, Certolizumab pegol, Clenoliximab, Daclizumab, Denosumab, Eculizumab, Edobacomab, Efalizumab, Erlizumab, Fezakinumab, Fontolizumab, Fresolimumab, Gantenerumab, Gavilimomab, Golimumab, Gomiliximab, Infliximab, Inolimomab, Keliximab, Lebrikizumab, Lerdelimumab, Mepolizumab,
Metelimumab, Muromonab-CD3, Natalizumab, Ocrelizumab, Odulimomab, Omalizumab,
Otelixizumab, Pascolizumab, Priliximab, Reslizumab, Rituximab, Rontalizumab, Rovelizumab, Ruplizumab, Sifalimumab, Siplizumab, Solanezumab, Stamulumab, Talizumab, Tanezumab, Teplizumab, Tocilizumab, Toralizumab, Ustekinumab, Vedolizumab, Vepalimomab,
Visilizumab, Zanolimumab, and Zolimomab aritox. In yet another embodiment, such therapeutic antibodies include, but are not limited to adalimumab, basiliximab, certolizumab pegol, eculizumab, efalizumab, infliximab, muromonab-CD3, natalizumab, and omalizumab. Alternately the therapeutic antibody can include abciximab or ranibizumab. Generally a therapeutic antibody is non-conjugated, or is conjugated with a radionuclide, cytokine, toxin, drug-activating enzyme or a drug-filled liposome.
Akt inhibitors include l L6-Hydroxymethyl-chiro-inositol-2-(R)-2-0-methyl-3-0- octadecyl-stt-glycerocarbonate, SH-5 (Calbiochem Cat. No. 124008), SH-6 (Calbiochem Cat. No. Cat. No. 124009), Calbiochem Cat. No. 12401 1 , Triciribine (NSC 154020, Calbiochem Cat. No. 124012), 10-(4'-(N-diethylamino)butyl)-2-chlorophenoxazine, Cu(II)Cl2(3-Formylchromone thiosemicarbazone), l ,3-dihydro- l -( l -((4-(6-phenyl-l H-imidazo[4,5-g]quinoxalin- 7-yl)phenyl)methyl)-4-piperidinyl)-2H-benzimidazol-2-one, GSK690693 (4-(2-(4-amino- 1 ,2,5- oxadiazol-3-yl)- l -ethyl-7-{ [(3S)-3-piperidinylmethyl]oxv 1 H-imidazo[4,5-c]pyridin-4-yl)-2- methyl-3-butyn-2-ol), SRI 3668 ((2, 10-dicarbethoxy-6-methoxy-5,7-dihydro-indolo[2,3-b] carbazole), GSK2141795, Perifosine, GSK21 1 10183, XL418, XL 147, PF-04691502, BEZ-235 [2-Methyl-2-[4-(3-methyl-2-oxo-8-quinolin-3-yl-2,3-dih^
phenyl]-propionitrile], PX-866 ((acetic acid (1 S,4E, 10R, 1 l R, 13S, 14R)-[4- diallylaminomethylene-6-hydroxy-l -methoxymethyl-10, 13-dimethy 1-3,7, 17-trioxo-
1 ,3,4,7, 10, 1 1 , 12, 13, 14, 15, 16, 17-dodecahydro-2-oxa-cyclopenta[a]phenanthren- l 1 -yl ester)), D-106669, CAL-101 , GDC0941 (2-( l H-indazol-4-yl)-6-(4-methanesulfonyl-piperazin-l - ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine), SF 1 126, SF 1 188, SF2523, TG I 00- 1 15 [3- [2,4-diamino-6-(3-hydroxyphenyl)pteridin-7-yl]phenol]. A number of these inhibitors, such as, for example, BEZ-235, PX-866, D 106669, CAL- 101 , GDC0941 , SF 1 126, SF2523 are also identified in the art as PI3 /mTOR inhibitors; additional examples, such as PI- 103 [3-[4-(4- morpholinylpyrido[3',2':4,5]furo[3,2-d]pyrimidin-2-yl]phenol hydrochloride] are well-known to those of skill in the art. Additional well-known PI3K inhibitors include LY294002 [2-(4- morpholinyl)-8-phenyl-4H-l -benzopyran-4-one] and wortmannin. mTOR inhibitors known to those of skill in the art include temsirolimus, deforolimus, sirolimus, everolimus, zotarolimus, and biolimus A9. A representative subset of such inhibitors includes temsirolimus, deforolimus, zotarolimus, and biolimus A9.
HDAC inhibitors include (i) hydroxamic acids such as Trichostatin A, vorinostat (suberoylanilide hydroxamic acid (SAHA)), panobinostat (LBH589) and belinostat (PXD101 ) (ii) cyclic peptides, such as trapoxin B, and depsipeptides, such as romidepsin (NSC 630176),
(iii) benzamides, such as MS-275 (3-pyridylmethyl-N-{4-[(2-aminophenyl)-carbamoyl]-benzyl}- carbamate), CI994 (4-acetylamino-N-(2aminophenyl)-benzamide) and MGCD0103 (N-(2- aminophenyl)-4-((4-(pyridin-3-yl)pyrimidin-2-ylamino)methyl)benzamide), (iv) electrophilic ketones, (v) the aliphatic acid compounds such as phenylbutyrate and valproic acid.
Hsp90 inhibitors include benzoquinone ansamycins such as geldanamycin, 17-DMAG
(17-Dimethylamino-ethylamino-17-demethoxygeldanamycin), tanespimycin (17-AAG, 17- allylamino- 17-demethoxygeldanamycin), EC5, retaspimycin (IPI-504, 1 8,21 -didehydro-17- demethoxy-18,21 -dideoxo- 18,21 -dihydroxy-17-(2-propenylamino)-geldanamycin), and herbimycin; pyrazoles such as CCT 018159 (4-[4-(2,3-dihydro- l ,4-benzodioxin-6-yl)-5-methyl- l H-pyrazol-3-yl]-6-ethyl- l ,3-benzenediol); macrolides, such as radicocol; as well as BIIB021 (CNF2024), SNX-5422, STA-9090, and AUY922. Miscellaneous agents include altretamine, arsenic trioxide, gallium nitrate, hydroxyurea, levamisole, mitotane, octreotide, procarbazine, suramin, thalidomide, lenalidomide, photodynamic compounds such as methoxsalen and sodium porfimer, and proteasome inhibitors such as bortezomib.
Biologic therapy agents include: interferons such as interferon-a2a and interferon-a2b, and interleukins such as aldesleukin, denileukin diftitox, and oprelvekin.
In addition to these anticancer agents intended to act against cancer cells, combination therapies including the use of protective or adjunctive agents, including: cytoprotective agents such as armifostine, dexrazonxane, and mesna, phosphonates such as parmidronate and zoledronic acid, and stimulating factors such as epoetin, darbepoetin, filgrastim, PEG-filgrastim, and sargramostim, are also envisioned.
Preparation and Examples
The compounds of the present invention can be synthesized using methods, techniques, and materials known to those of skill in the art, such as described, for example, in March, ADVANCED ORGANIC CHEMISTRY 4th ed., (Wiley 1992); Carey and Sundberg,
ADVANCED ORGANIC CHEMISTY 3rd ed., Vols. A and B (Plenum 1992), and Greene & Wuts, "Protective Groups in Organic Synthesis," Wiley Interscience, 1999. Starting materials useful for preparing compounds of the invention and intermediates thereof are commercially available from sources, such as Aldrich Chemical Co. (Milwaukee, Wis.), Sigma Chemical Co. (St. Louis, Mo.), Maybridge (Cornwall, England), Asinex (Winston-Salem, NC), ChemBridge (San Diego, CA ), ChemDiv (San Diego, CA), SPECS (Delft, The Netherlands), Timtec (Newark, DE), or alternatively can be prepared by well-known synthetic methods (see, e.g. , Harrison et al., "Compendium of Synthetic Organic Methods", Vols. 1 -8 (John Wiley and Sons, 1971 - 1996); "Beilstein Handbook of Organic Chemistry," Beilstein Institute of Organic Chemistry, Frankfurt, Germany; Feiser et al., "Reagents for Organic Synthesis," Volumes 1 -21 , Wiley Interscience; Trost et al., "Comprehensive Organic Synthesis," Pergamon Press, 1991 ; "Theilheimer's Synthetic Methods of Organic Chemistry," Volumes 1 -45, Karger, 1991 ; March, "Advanced Organic Chemistry," Wiley Interscience, 1991 ; Larock "Comprehensive Organic Transformations," VCH Publishers, 1989; Paquette, "Encyclopedia of Reagents for Organic Synthesis," 3rd edition, John Wiley & Sons, 1995). Other methods for synthesis of the present compounds and/or starting materials thereof are either described in the art or will be readily apparent to the skilled artisan. Alternatives to the reagents and/or protecting groups may be found in the references provided above and in other compendiums well known to one skilled in the art.
Preparation of the present compounds may include one or more steps of protection and deprotection (e.g. , the formation and removal of acetal groups). Guidance for selecting suitable protecting groups can be found, for example, in Greene & Wuts, "Protective Groups in Organic Synthesis," Wiley Interscience, 1999. In addition, the preparation may include various purifications, such as column chromatography, flash chromatography, thin-layer chromatography (TLC), recrystallization, distillation, high-pressure liquid chromatography (HPLC) and the like. Also, various techniques well known in the chemical arts for the identification and quantification of chemical reaction products, such as proton and carbon-13 nuclear magnetic resonance(Ή and l3C NMR), infrared and ultraviolet spectroscopy (IR and UV), X-ray crystallography, elemental analysis (EA), HPLC and mass spectroscopy (MS) can be used as well. The preparation may also involve any other methods of protection and deprotection, purification and identification and quantification that are well known in the chemical arts.
The following examples illustrate and do not limit the invention.
Example 1. Synthesis of (ZV 5-((5-((3-chlorophenyiymethyl')amino')-7- ("cyclopropylamino'tpyrazolori .S-alpyrimidin^-ynmethylenelimidazolidine^^-dione
Figure imgf000211_0001
(Z)-5-((5-Chloro-7-(cyclopropylamino)pyrazolo[l ,5-a]pyrimidin-3- yl)methylene)imidazolidine-2,4-dione (75 mg, 0.23 mmol) was suspended in 1 ,4-dioxane (1 .6 mL). 3-Chloro-N-methylaniline (49 mg, 0.35 mmol), Cs2C03 (105 mg, 0.32 mmol), (±)-ΒΓΝΑΡ (9 mg, 0.06 mmol) and palladium(II) acetate (7 mg, 0.04 mmol) were then added. The mixture was sealed and irradiated at 130 °C for 40 min in the microwave. H2O (8 mL) was added and the precipitate was filtered off and dried. The crude residue was purified via flash column chromatography (2.5% MeOH/dichloromethane) to yield (Z)-5-((5-((3- chlorophenyl)(methyl)amino)-7-(cyclopropylamino)pyrazolo[l ,5-a]pyrimidin-3- yl)methylene)imidazolidine-2,4-dione (8 mg, 8%) as a bright yellow solid. LCMS (ES): >90% pure, m/z 424 [M+H]+.
The molecules described in the following table (Table 1 A) were prepared using the procedures similar to those exemplified in Example 1. All compounds were characterized by LCMS. Table I B shows the biological activities of the compounds listed in Table 1A as well as Examples 1 , 2, 4, 6, 7, 10, and 12. That is, Compounds 1A to IO in Table I B are the compounds listed in Table 1 A and Examples 1 , 2, 4, 6, 7, 10, and 12.
Table 1 A.
Figure imgf000212_0001
Figure imgf000213_0001
Example 2. Synthesis of (Z)-5-((5-(3-Chloro-4-Cpiperazin-l -yl)phenvh
('cvclopropylamino')pyrazolo[1.5-a]pyrimidin-3-yl')methylene)imidazolidine-2,4-dione 2,2,2- trifluoroacetate
Figure imgf000213_0002
(Z)-7¾/Y-butyl 4-(2-chloro-4-(7-(cyclopropylamino)-3-((2,5-dioxoimidazoIidin-4- ylidene)methyl)pyrazolo[l ,5-a]pyrimidin-5-ylamino)phenyl)piperazine-l -carboxylate (22 mg, 0.037 mmol) was dissolved in dichloromethane (0.5 mL) and trifluoroacetic acid (0.5 mL). After one hour, the solution was concentrated under a stream of air. The crude residue was triturated with Et20 (4 mL) and filtered to yield (Z)-5-((5-(3-chloro-4-(piperazin-l-yl)phenylamino)-7- (cyclopropyIamino)pyrazolo[l ,5-a]pyrimidm^ 2,2,2- trifluoroacetate (18 mg, 80%). LCMS (ES): >90% pure, m/z 494 [M+H]+.
Example 3. Synthesis of 7-(cyclopropylamino)-5-(2,5-dichlorophenylamino")pyrazolon .5- alpyrimidine-3-carbaldehyde
Figure imgf000214_0001
In a microwave reaction tube, 2,4-dichloroaniline (190 mg, 1.2 mmol), tert-butyl 5- chloro-3-formylpyrazolo[l ,5-a]pyrimidin-7-yl(cyclopropyl)carbamate (340 mg, 1 mmol), tris(dibenzylideneacetone)dipalladium(0), X-Phos Ligand (1 1 mg, 0.02 mmol), and LiHMDS (1M in THF, 2.2 mL, 2.2 mmol) were added. The reaction was heated at 65°C in the microwave for 1 hour then solvent was removed by evaporation under a stream of nitrogen. The residue was taken up in DCM, filtered, and concentrated under vacuum. The crude material was then dissolved in (1 : 1 ) TFA/DCM (3 mL) and stirred at room temperature for 1 hour. Excess solvent and TFA were removed by evaporation under a stream of nitrogen. The residue was taken up in DCM, washed with saturated NaHCC solution (3x) and saturated NaCl solution. The organic layer was isolated, dried over anhydrous MgSC , filtered, and evaporated to dryness. The residue was purified by column chromatography on silica using 30% EtOAc/hexanes as the eluent. The pure fractions were combined and the solvent was removed. This material was washed with 20% MeOH/DCM to provide 7-(cyclopropylamino)-5-(2,5- dichlorophenylamino)pyrazolo[l ,5-a]pyrimidine-3-carbaldehyde (67 mg, 18% yield). LCMS (M+ 1 =362) Example 4. Synthesis of 5-((7-(cvclopropylaminoy5-(2,5-dichlorophenylamino pyrazolo[1.5- alpyrimidin-3-vDmethylene)imidazolidine-2,4-dione
Figure imgf000215_0001
Hydantoin (20 mg, 0.2 mmol) and 3-chloro-4-(7-(cyclopropylamino)-3-formyl-6- methylpyrazolo[l ,5-a]pyrimidin-5-ylamino)benzonitrile (30 mg, 0.1 mmol) were dissolved in ethanol (0.5 mL) along with piperidine (20 uL, 0.2 mmol). The reaction was heated at 85 °G in the microwave for 2 hours. The reaction was cooled to r.t., diluted with water, and the precipitate was collected and washed with water, (1 : 1 ) ethanol/water, then ethanol. The solid was dried in vacuo to give 5-((7-(cyclopropylamino)-5-(2,5-dichlorophenylamino)pyrazolo[l ,5- a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (35 mg, 94% yield). LCMS (M=444)
Example 5. Synthesis of 4-(7-(cvclopropylamino)-3-formylpyrazolori ,5-alpyrimidin-5- ylamino>3-methylbenzonitrile
Figure imgf000215_0002
Same procedure as Example 3. LCMS m/z 333 [M+H]+
Example 6. Synthesis of 4-(7-(cyclopropylamino)-3-((2,5-dioxoimidazolidin-4- ylidene")methynpyrazolo[l ,5-alpyrimidin-5-ylamino)-3-methylbenzonitrile
Figure imgf000216_0001
Same procedure as Example 4. LCMS m/z 415 [M+H]+ Example 7. Synthesis of (Z)-5-('('5-('4-aminophenylaminoV7-(cvclopropylamino')pyrazolon ,5- fllpyrimidin-3-y0methylene)imidazolidine-2,4-dione
Figure imgf000216_0002
(Z)-5-((7-(Cyclopropylamino)-5-(4-hitrophenylamino)pyrazolo[l ,5-o]pyrimidin-3- yl)methylene)imidazolidine-2,4-dione (1 1 mg, 0.026 mmol) was suspended in MeOH (0.5 mL) and Pd/C (10%, Degussa type, 10 mg) was added. ¾ was bubbled through the solution for ~10 seconds, and the reaction was kept under a balloon of ¾. After 1 .5 h, the solution was filtered over a pad of celite eluting with 50% MeOH/dichloromethane. The filtrate was concentrated in vacuo and the residue was purified via preparative TLC (5% MeOH/dichloromethane) to provide (Z)-5-((7-(cyclopropylamino)-5-(4-nitrophenylamino)pyrazolo[l ,5-a]pyrimidin-3- yl)methylene)imidazolidine-2,4-dione (8 mg, 78%) as a bright yellow solid. LCMS (ES): >90% pure, m/z 391 [M+H]+. Example 8. Synthesis of /e -butyl cyclopropyl(3-formyl-5-(3-(trifluoromethyl")pyridin-4- ylamino)pyrazolon ,5-alpyrimidin-7-yl)carbamate
Figure imgf000217_0001
7e /-butyl 5-chloro-3-foirnylpyrazolo[l ,5-a]pyrimidin-7-yl(cyclopropyl)carbarnate (1 12 mg, 0.33 mmol) was suspended in 1 ,4-dioxane (2.2 mL). 4-amino-3-(trifluromethyl)pyridine (80 mg, 0.50 mmol), Cs2C03 (151 mg, 0.46 mmol), (±)-ΒΓΝΑΡ (12 mg, 0.02 mmol) and palladium(II) acetate (9 mg, 0.013 mmol) were then added. The mixture was sealed and irradiated at 1 10 °C for 20 min in the microwave. Et20 (8 mL) was added and the filtrate was concentrated in vacuo to a brown foam. The crude residue was purified via flash column chromatography (45-60% EtOAc/hexanes) to yield teri-butyl cyclopropyl(3-formyl-5-(3-
(trifluoromethyl)pyridin-4-ylamino)pyrazolo[l ,5-a]pyrimidin-7-yl)carbamate (72 mg, 47%) as a yellow foam. LCMS (ES): >90% pure, m/z 463 [M+H]+.
Example 9. Synthesis of 7-(cvclopropylamino)-5-(3-('trifluoromethyl')pyridin-4- ylamino)pyrazolo[1 ,5-fl1pyrimidine-3-carbaldehyde 2,2,2-trifluoroacetate
7¾r/-butyl cyclopropyl(3-formyl-5-(3-(trifluoromethyl)pyridin-4-ylamino)pyrazolo[l ,5- a]pyrimidin-7-yl)carbamate (72 mg, 0.16 mmol) was dissolved in dichloromethane (0.7 mL) and trifluoroacetic acid (0.7 mL). After one hour, the solution was concentrated under a stream of air. The crude residue was triturated with Et20 (4 mL) and filtered to furnish 7- (cyclopropylamino)-5-(3-(trifluoromethyl)pyridin-4-ylamino)pyrazolo[l ,5-a]pyrimidine-3- carbaldehyde 2,2,2-trifluoroacetate (41 mg, 55%). LCMS (ES): >90% pure, m/z 363 [M+H]+.
Figure imgf000218_0001
7-(Cyclopropylamino)-5-(3-(trifluoromethyl)pyridin-4-ylamino)pyrazolo[ l ,5- a]pyrimidine-3-carbaldehyde 2,2,2-trifluoroacetate (20 mg, 0.055 mmol) was suspended in EtOH (0.5 mL). Hydantoin (7 mg, 0.07 mmol) and piperidine (18 μί, 0.18 mmol) were added and the reaction was heated to 80 °C. After 18 h, the solution was filtered while warm and the filter cake was washed with warm EtOH (3 mL) to afford (Z)-5-((7-(cyclopropylamino)-5-(3- (trifluoromethyl)pyridin-4-ylamino)pyrazolo[l ,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4- dione (9 mg, 50%) as a bright yellow solid. LCMS (ES): >95% pure, m/z 445 [M+H]+. Example 1 1 . Synthesis of ethyl 7-('2-chloro-4-(7-('cvclopropylamino')-3-formylpyrazolo[1.5-
Figure imgf000218_0002
To 5-(3-chloro-4-hydroxyphenylamino)-7-(cyclopropylamino)pyrazolo[l ,5-a]pyrimidine- 3-carbaldehyde (300 mg, 0.87 mmol) in DMF (8 mL) was added K2C03 (301 mg, 2.18 mmol). The mixture was cooled to 0°C and ethyl-7-bromoheptanoate (217 mg, 0.92 mmol) was added slowly and stirred while warming to rt overnight. Diluted with EtOAc and washed I X with H2O, 3X with brine. The organic layer was dried with MgS0 , filtered, and the solvent was removed by rotary evaporation. Dried under vacuum to provide 440 mg (100%) of ethyl 7-(2-chloro-4-(7- (cyclopropylamino)-3-formylpyrazolo[ l ,5-a]pyrimidin-5-ylamino)phenoxy)heptanoate. LCMS (ES): >95% pure, m/z 500 [M+l ]+. Example 12. Synthesis of ("ZVethyl 7-f2-chloro-4-('7-(cvclopropylamino')-3-('('2.5- dioxoimidazolidin-4-ylidene methvnpyrazolo[1.5-alpyrimidin-5-ylamino)phenoxy)heptanoate
Figure imgf000219_0001
To ethyl 7-(2-chloro-4-(7-(cyclopropylamino)-3-formylpyrazolo[l ,5-a]pyrimidin-5- ylamino)phenoxy)heptanoate (440 mg, 0.88 mmol) in EtOH (10 mL) was added hydantoin (132 mg, 1.32 mmol) followed by piperidine (130 xL, 1.32 mmol). The reaction mixture was stirred at 80°C for 3h, then cooled to rt and diluted with 5 mL of H2O. The resulting precipitate was filtered off, rinsed with H20 followed by 1 : 1 H20/EtOH, and dried under vacuum to provide 190 mg of (Z)-ethy 1 7-(2-chloro-4-(7-(cyclopropylamino)-3-((2,5-dioxoimidazolidin-4- ylidene)methyl)pyrazolo[l ,5-a]pyrimidin-5-ylamino)phenoxy)heptanoate. (LCMS (ES): >95% pure, m/z 582 [M+H]+.
Table I B.
Figure imgf000219_0002
Figure imgf000220_0001
Example 13. Synthesis of (E)-N5-('2-bromo-5-fluorobenzyl')-3-(('2-bromo-5- fluorobenzylimino')methvn-N7-cvclopropylpyrazolofl ,5-alpyrimidine-5.7-diamine
Figure imgf000220_0002
To a suspension of tert-butyl 5-chloro-3-formylpyrazolo [1 , 5-a] pyrimidin-7-yl
(cyclopropyl) carbamate (1.0 g, 3.0 mmol) in isopropanol (5 mL), was added 2-bromo 5-fluoro benzyl amine hydrochloride (1.4 g, 6.0 mmol) and DIPEA (0.6 mL). The reaction mixture was stirred at 80°C for 48 hours, then cooled to room temperature and diluted with water. The yellow precipitate was stirred in ethyl acetate for 30 minutes, filtered, and dried under vacuum. To the crude product, was added HCl/dioxane (3.0 mL) and.the mixture was stirred at 80°C for overnight. The resulting brown solid was filtered and dried under vacuum to provide, (E)-N5-(2- bromo-5-fluorobenzyl)-3-((2-bromo-5-fluoroben2ylimino)methyl)-N7-cyclopropylpyrazolo [1 , 5-a] pyrimidine-5, 7-diamine ( 1 .2 g). (LCMS (ES): >90% pure, m/z 591 [M+H]+.
Example 14. Synthesis of (Z)-5-(f5-(2-bromo-5-fluorobenzylarnino)-7- (cyclopropylamino')pyrazolori .5-a1pyrimidin-3-yl')methylene)imidazolidine-2.4-dione
Figure imgf000221_0001
To (E)-N5-(2-bromo-5-fluorobenzyl)-3-((2-bromo-5-fluorobenzylimino) methyl)-N7- cyclopropylpyrazolo [1 , 5-a] pyrimidine-5, 7-diamine ( 1 .2 g, 2.0 mmol) in EtOH (4 mL), was added hydantoin (384 mg, 4.0 mmol) and pyrrolidine (0.4 mL, 4.0 mmol). The reaction mixture was stirred at 80°C for three hours, then cooled to room temperature and the resulting yellow precipitate was filtered, washed with ethanol to provide (Z)-5-((5-(2-bromo-5- fluorobenzylamino)-7-(cyclopropylamino)pyrazolo[l ,5-a]pyrimidin-3-yl)methylene) imidazolidine-2,4-dione (785 mg, 90% yield). (LCMS (ES): >95% pure, m z 486 [M+H]+.
Example 1 5. Synthesis of (Z)-5-((7-(cvclopropylamino)-5-(5-fluoro-2-(thiophen-2- yl)benzylamino)pyrazolori .5-alpyrimidin-3-yl)methylene)imidazolidine-2.4-dione
Figure imgf000221_0002
To a suspension of (Z)-5-((5-(2-bromo-5-fIuorobenzylamino)-7-(cyclopropylamino) pyrazolo [1 , 5-a] pyrimidin-3-yl) methylene) imidazolidine-2, 4-dione (20 mg, 0.05 mmol) in a mixture of DMF/water (1 mL), were added thiophene-2-boronic acid (8.0 mg, 0.06 mmol), cesium carbonate (40 mg, 0.12 mmol) and PdC dppf (2.0 mg, O.01 mmol). The mixture was sealed and heated at 120 °C for 15 minutes in the microwave. The reaction mixture was cooled to room temperature and diluted with water. The resulting brown solid was then diluted with 1 : 1 mixture of MeOH and dichloromethane and was purified using preparative TLC to provide (Z)- 5-((7-(cyclopropylamino)-5-(5-fluoro-2-(thiophen-2-yl)benzylamino)pyrazolo[l ,5-a]pyrimidin- 3-yl)methy lene)imidazolidine-2,4-dione. LCMS (ES): >95% pure, m/z 490 [M+H]+.
Figure imgf000222_0001
Same procedure as Example 3. LCMS (M+ 1 =485)
Example 17. Synthesis of (Z)-5-((5-((2'-chloro-4-fluorobiphenyl-2-yl)methylamino')-7- (cyclopropylamino pyrazolori ,5-alpyrimidin-3-l)methylene')imidazolidine-2,4-dione
Figure imgf000223_0001
To a suspension of (Z)-5-((5-(2-bromo-5-fluorobenzylamino)-7-(cyclopropylamino) pyrazolo [1 , 5-a] pyrimidin-3-yl) methylene) imidazolidine-2, 4-dione (15 mg, 0.03 mmol) in DMF (0.2 mL), was added 2-chlorophenylboronic acid (7 mg, 0.05 mmol) and sodium acetate (12.5 mg, 0.1 mmol). The mixture was degassed under nitrogen for 3 minutes, then PdC dppf (1.3 mg, <0.01 mmol) was added. The reaction mixture was heated in microwave for 10 minutes at 1 10 °C, diluted with 1 : 1 mixture of MeOH and dichloromethane and purified using preparative TLC to provide (Z)-5-((5-((2'-chloro-4-fluorobiphenyl-2-yl)methylamino)-7- (cyclopropylamino)pyrazolo[l ,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione as solid. LCMS (ES) :> 95% pure, m/z 518 [M+H]+.
Figure imgf000223_0002
Same procedure as Example 5. LCMS (M+l =484)
The compounds described in the following Table 2A were prepared using procedures similar to those exemplified in Example 17. All compounds were characterized by LCMS. Table 2B shows the biological activities of the compounds listed in Table 2A and Examples 14 to 18. That is, Compounds I P to 1 Z are compounds listed in Table 2A and Examples 14 to 18. Table 2A.
Figure imgf000224_0001
Table 2B. AB: BxPC3 IC50
Compound CK2: IC50 (μΜ) AB: MDAMB453 IC50 (μΜ)
(μΜ)
IP >1
iQ <0.1
1R <1
IS <1
IT <1
1U <1
IV >1
1W >1
IX <1
1Y <0.1
1Z <1
Example 19. Synthesis of (EVN5-r2-bromo-4-fluorobenzylV3-((2-bromo-4- fluorobenzylimino')methvn-N7-cvclopropylpyrazolon,5-alpyrimidine-5.7-diamine
Figure imgf000225_0001
To a suspension of tert-butyl 5-chloro-3-formylpyrazolo [1, 5-a] pyrimidin-7-yl (cyclopropyl) carbamate (2.0 g, 6.0 mmol) in isopropanol (5 mL), was added 2-bromo 4-fluoro benzyl amine hydrochloride (3.0g, 12.0 mmol) and DIPEA (1.2 mL). The reaction mixture was stirred at 80°C for 48 hours, then cooled to room temperature and diluted with water. The yellow precipitate was stirred in ethyl acetate for 30 minutes, filtered, and dried under vacuum. To the crude product, was added HCl/dioxane (3 mL) and the mixture was stirred at 80°C for overnight. The resulting white precipitate was filtered and dried under vacuum to provide, (E)- N5-(2-bromo-4-fluorobenzyl)-3-((2-bromo-4-fluorobenzylimino)methyl)-N7- cyclopropylpyrazolo[l ,5-a]pyrimidine-5,7-diamine (771 mg). (LCMS (ES): >90% pure, m/z 591 [M+H]+.
Example 20. Synthesis of (Z)-5-((5-(2-bromo-4-fluorobenzylaminoV7-
(cyclopropylamino')pyrazolo l ,5-alpyrimidin-3-yl')methylene)imidazolidine-2.4-dione
Figure imgf000226_0001
To (E)-N5-(2-bromo-4-fluorobenzyl)-3-((2-bromo-4-fluorobenzylimino) methyl)-N7- cyclopropylpyrazolo [1 , 5-a] pyrimidine-5, 7-diamine (771 mg, 1.3 mmol) in EtOH (2 mL), was added hydantoin (261 mg, 3.0 mmol) and pyrrolidine (0.3 mL, 3.0 mmol). The reaction mixture was stirred at 80°C for overnight, then cooled to room temperature and the resulting yellow precipitate was filtered, washed with ethanol to provide, (Z)-5-((5-(2-bromo-4- fluorobenzylamino)-7-(cyclopropylamino)pyrazolo[l ,5-a] pyrimidin-3-yl)methylene) imidazolidine-2,4-dione (548 mg, 90% yield). (LCMS (ES): >90% pure, m/z 486 [M+H]+.
Example 21 . Synthesis of (Z")-5-((7-(cvclopropylaminoV5-(4-fluoro-2-Cthiophen-3- vDbenzylamino'lpyrazolof KS-alpyrimidin-S-ynmethylene^imidazolidine^^-dione
Figure imgf000227_0001
To a suspension of (Z)-5-((5-(2-bromo-4-fluorobenzylamino)-7-(cyclopropylamino) pyrazolo [1 , 5-a] pyrimidin-3-yl) methylene) imidazolidine-2, 4-dione (10 mg, 0.02 mmol) in DMF (0.2 mL), was added thiophene-3-boronic acid (4 mg, 0.03 mmol) and sodium acetate (9 mg, 0.1 mmol). The mixture was degassed under nitrogen for 3 minutes, then PdC^dppf (0.8 mg, <0.01 mmol) was added. The reaction mixture was heated in microwave for 10 minutes at 120 °C, diluted with 1 : 1 mixture MeOH and dichloromethane and purified using preparative TLC to provide (Z)-5-((7-(cyclopropylamino)-5-(4-fluoro-2-(thiophen-3-yl)benzylamino)pyrazolo[ l ,5- a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione. LCMS (ES) :> 95% pure, m/z 490
[M+H]+.
Figure imgf000227_0002
Same procedure as Example 3. LCMS (M+l =490) The molecules described in the following table were prepared using chemistries similar to those exemplified in Example 21 . All compounds were characterized by LCMS. Table 3B shows the biological activities of the compounds listed in Table 3A.
Example 23. Synthesis of (Z)-5-((5-(2-(2-aminopyrimidin-5-yl)-4-fluorobenzylamino)-7- ('cvclopropylamino')pyrazolo 1.5-alpyrimidin-3-yl')methylene")imidazolidine-2.4-dione
Figure imgf000228_0001
To (Z)-5-((5-(2-bromo-4-fluorobenzylamino)-7-(cyclopropylamino)pyrazolo[l ,5-a] pyrimidin-3-yl)methylene) imidazolidine-2,4-dione (15 mg, 0.03 mmol) in DME/Ethanol (2: 1 ) was added 2-Aminopyrimidine-5-boronic acid (7 mg, 0.05 mmol) and 2M Na2 CO3 (2 drops). The mixture was degassed'under nitrogen for 3 minutes and then PdCl2dppf (0.8 mg, O.01 mmol) was added. The mixture was heated in microwave for 10 minutes at 120 °C. The reaction was then partitioned between ethyl acetate and water, the organic layer was dried under sodium sulfate concentrated, diluted with 1 : 1 mixture of MeOH and dichloromethane and was purified using preparative TLC to provide (Z)-5-((5-(2-(2-aminopyrimidin-5-yl)-4-fluorobenzylamino)-7- (cyclopropylamino)pyrazolo[l ,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione. LCMS (ES) :> 95% pure, m/z 501 [M+H]+. The compounds described in the following Table 3 A were prepared using procedures similar to those exemplified in the above examples. All compounds were characterized by LCMS. Table 3B shows the biological activities of the compounds listed in Table 3A and Examples 20 to 23. That is, Compounds I P to 1Z are compounds listed in Table 3A and Examples 20 to 23.
Table 3A.
Figure imgf000229_0001
Figure imgf000230_0001
Table 3B.
Figure imgf000230_0002
Example 24. Synthesis of (EVN5-(2-bromobenzylV3-(f2-bromobenzylimino)methy0-N7- cyclopropylpyrazolof l ,5-a1pyrimidine-5.7-diamine
Figure imgf000231_0001
To a suspension of tert-butyl 5-chloro-3-formylpyrazolo [1 , 5-a] pyrimidin-7-yl (cyclopropyl) carbamate (1.0 g, 3.0 mmol) in isopropanol (5 mL), was added 2-bromo benzyl amine hydrochloride (1.3 g, 6.0 mmol) and DIPEA (0.6 mL). The reaction mixture was heated at 80°C for 48 hours, then cooled to room temperature and diluted with water. The yellow precipitate was stirred in ethyl acetate for 30 minutes, filtered, and dried under vacuum. To the crude product, was added HCl/dioxane (3.0 mL) and the mixture was stirred at 80°C for overnight. The resulting brown precipitate was filtered and dried under vacuum to provide (E)- N5-(2-bromobenzyl)-3-((2-bromobenzylimino)methyl)-N7-cyclopropylpyrazolo[l,5-a] pyrimidine-5,7-diamine (743mg). (LCMS (ES): >90% pure, m/z 555 [M+H]+.
Example 25. Synthesis of (Z)-5-f(5-(2-bromobenzylamino)-7-(cvclopropylamino)pyrazolon ,5- alpyrimidin-3-yl")methylene imidazolidine-2.4-dione
Figure imgf000231_0002
To a suspension of (E)-N5-(2-bromobenzyl)-3-((2-bromobenzylimino) methyl)-N7- cyclopropylpyrazolo [1 , 5-a] pyrimidine-5, 7-diamine (743mg, 1.4 mmol) in EtOH (2 mL), was added hydantoin (260 mg, 3.0 mmol) and pyrrolidine (0.3 mL). The reaction mixture was stirred at 80°C for overnight, then cooled to room temperature and the resulting yellow precipitate was filtered, washed with ethanol to provide, (Z)-5-((5-(2-bromobenzylamino)-7-(cyclopropylamino) pyrazolo[l ,5-a] pyrimidin-3-yl)methylene) imidazolidine-2, 4-dione (498 mg, 80% yield). (LCMS (ES): >90% pure, m z 469 [M+H]+.
Figure imgf000232_0001
To a suspension of (Z)-5-((5-(2-bromobenzyIamino)-7-(cyclopropylamino)pyrazolo[ l ,5- a] pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (8 mg, 0.02 mmol) in DMF (0.2 mL), was added 1 -H pyrazole-4-boronic acid (4 mg, 0.03 mmol) and sodium acetate (9 mg, 0.1 mmol). The mixture was degassed under nitrogen for 3 minutes, and then PdC^dppf (0.8 mg, <0.01 mmol) was added. The reaction mixture was heated in the microwave for 10 minutes at 120 °C, diluted with 1 : 1 mixture of MeOH and dichloromethane and purified using preparative TLC to provide (Z)-5-((5-(2-(l H-pyrazol-4-yl)benzylamino)-7-(cyclopropylamino)pyrazolo[l ,5- a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione. (LCMS (ES): >95% pure, m/z 456
[M+H]+.
Example 27. Synthesis of (Z)-5-('(7-('cvcloproDylamino)-5-r(3'-('trifluoromethyl')biphenyl-2- ynmethylamino)pyrazolo|" 1.5-a1pyrimidin-3-vnmethylene')imidazolidine-2,4-dione
Figure imgf000233_0001
To (Z)-5-((5-(2-bromo-4-fluorobenzylamino)-7-(cyclopropylamino) pyrazolo [1 , 5-a] pyrimidin-3-yl) methylene) imidazolidine-2, 4-dione (10 mg, 0.02 mmol) in DME: Ethanol (2: 1 ), was added 3-trifluoromethyphenyllboronic acid (6 mg, 0.03 mmol) and 2M Na2 CO3 (2 drops). The mixture was degassed under nitrogen for 3 minutes and then PdC dppf (0.8 mg, O.01 mmol) was added. The mixture was heated in microwave for 10 minutes at 1 10 °C. The reaction was then partitioned between ethyl acetate and water, the organic layer was dried under sodium sulfate concentrated, diluted with 1 : 1 mixture of MeOH and dichloromethane and was purified using preparative TLC to yield (Z)-5-((7-(cyclopropylamino)-5-((3'-(trifluoromethyl)biphenyl-2- yl)methylamino)pyrazolo[l ,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione. (LCMS (ES): >95% pure, m/z 534 [M+H]+.
The compounds described in the following Table 4A were prepared using procedures similar to those exemplified in Examples 26 and 27. All compounds were characterized by LCMS. Table 4B shows the biological activities of the compounds listed in Table 4A as well as Examples 25 to 27.
Table 4A.
Figure imgf000233_0002
Figure imgf000234_0001
Figure imgf000235_0001
Table 4B.
Figure imgf000235_0003
Figure imgf000235_0002
alpyrimidine-3-carbaldehyde
Figure imgf000236_0001
To a suspension of 5-chloro-7-(cyclopropylamino) pyrazolo [1 , 5-a] pyrimidine-3- carbaldehyde (500 mg, 2.0 mmol) in DMF (4 mL), was added 1 -Boc-piperazine (1.2 g, 6.0 mmol), potassium carbonate (0.5 g, 4.0 mmol) and DIPEA (0.4 mL). The reaction mixture was stirred at 80°C for overnight, then cooled to room temperature and diluted with water. The resulting white precipitate was filtered and dried under vacuum to provide tert-butyl 4-(7- (cyclopropylamino)-3-formylpyrazolo [1 , 5-a] pyrimidin-5-yl) piperazine-l -carboxylate (655mg). LCMS (ES): >90% pure, m/z 387 [M+H]+.
Example 29. Synthesis of (ZV5-((7-(cvclopropylamino)-5-(piperazin-l -vnpyrazolo l ,5- alpyrimidin-3-yl")methylene)imidazolidine-2,4-dione
Figure imgf000236_0002
To a suspension of tert-butyl 4-(7-(cyclopropylamino)-3-formylpyrazolo [1 , 5-a] pyrimidin-5-yl) piperazine-l -carboxylate (403 mg, 1 .0 mmol) in EtOH (2 mL), was added hydantoin (208 mg, 2.1 mmol) and piperdine (0.2 mL). The reaction mixture was heated to 80°C for 3 hours, then cooled to room temperature and the resulting yellow precipitate was filtered, washed with ethanol to provide (Z)-5-((7-(cyclopropylamino)-5-(piperazin- l -yl) pyrazolo [1 , 5- a] pyrimidin-3-yl) methylene) imidazolidine-2, 4-dione (410 mg, 95% yield). LCMS (ES): >95% pure, m/z 369 [M+H]+.
Example 30. Synthesis of (Z)-methyl 4-(7-("cvclopropylamino)-3-("("2.5-dioxoimidazolidin-4- ylidene)methvnpyrazolori .5-a1pyrimidin-5-yl")piperazine-l -carboxylate
Figure imgf000237_0001
To a suspension of (Z)-5-((7-(cyclopropylamino)-5-(piperazin- l -yl)pyrazolo [1 ,5-a] pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (10 mg, 0.02 mmol) in THF (0.4 mL), was added methyl chloro formate (4 μί, 0.03 mmol) and DIPEA (5.6 μΕ). The reaction mixture was stirred for half hour at room temperature and the resulting white precipitate was filtered, washed with ethyl acetate to provide (Z)-methyl 4-(7-(cyclopropylamino)-3-((2,5-dioxoimidazolidin-4- ylidene)methyl)pyrazolo[l ,5-a]pyrimidin-5-yl)piperazine- l -carboxylate. LCMS (ES): >95% pure, m/z 427 [M+H]+.
Example 31 . Synthesis of (Z)-5-((7-(cvclopropylamino)-5-(4-(2-fluorobenzoyl)piperazin-l - vnpyrazolorKS-alpyrimidin-S-vnmethylene midazolidine^^-dione
Figure imgf000237_0002
To a suspension of (Z)-5-((7-(cyclopropylamino)-5-(piperazin- l -yl)pyrazolo [1 ,5-a] pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (10 mg, 0.02 mmol) in THF (0.4 mL), added 2-fluoro benzoyl chloride (8.5 mg, 0.05 mmol) and DIPEA (5.6 D L). The reaction mixture was stirred for half hour at room temperature and the resulting white precipitate was filtered, washed with ethyl acetate to provide (Z)-5-((7-(cyclopropylamino)-5-(4-(2-fluorobenzoyl)piperazin-l - yl)pyrazolo[l ,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (410 mg, 95% yield). LCMS (ES): >95% pure, m/z 491 [M+H]+.
The compounds described in the following Table 5A were prepared using procedures similar to those exemplified in Example 31 . All compounds were characterized by LCMS. Table 5D shows the biological activities of the compounds listed in Table 5A, 5B, and 5C as well as Examples 29 to 31 , 32, and 33.
Table 5A.
Figure imgf000238_0001
Example 32. Synthesis of (Zy5-f(7-(cyclopropylamino)-5-(4-isobutylpiperazin- l - vOpyrazolon ,5-alpyrimidin-3-yl)methylene)imidazolidine-2,4-dione
Figure imgf000239_0001
To a suspension of (Z)-5-((7-(cyclopropylamino)-5-(piperazin- l -yI)pyrazolo [1 ,5-a] pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (10 mg, 0.03 mmol) in THF (0.2 mL), was added acetic acid (2 drops), isobuteraldehyde (9.4 μΐ,, 0.1 mmol) and sodium
triacetoxyborohydride (25 mg, 0.12 mmol). The reaction mixture was then stirred at room temperature for half hour, diluted with MeOH, and prepared by HPLC purification to provide (Z)-5-((7-(cyclopropylamino)-5-(4-isobutylpiperazin-l -yl) pyrazolo [1 , 5-a] pyrimidin-3-yl) methylene) imidazolidine-2, 4-dione. LCMS (ES): >95% pure, m z 425 [M+H]+.
The molecules described in the following table were prepared using chemistries
similar to those exemplified in Example 32. All compounds were characterized by LCMS. Table 5D shows the biological activities of the compounds listed in Table 5B.
Table 5B.
Figure imgf000239_0002
Figure imgf000240_0001
Example 33. Synthesis of (Z)-5-((7-(cyclopropylamino)-5-(4-(2,4- difluorophenylsulfonvDpiperazin- l -vDpyrazolori .S-alpyrimidin^-ynmethylene^imidazolidine- 2,4-dione
Figure imgf000240_0002
To a suspension of (Z)-5-((7-(cyclopropylamino)-5-(piperazin-l -yl) pyrazolo [1 , 5-a] pyrimidin-3-yl) methylene) imidazolidine-2, 4-dione (10 mg, 0.03 mmol) in THF (0.2 mL), was added 2, 4 difluoro benzene sulfonyl chloride (1 1 mg, 0.05 mmol) and triethylamine (0.5 mL). The mixture was stirred at room temperature for half hour under nitrogen, diluted with DMSO and prepared by HPLC purification to provide (Z)-5-((7-(cyclopropylamino)-5-(4-(2,4- difluorophenylsulfonyl)piperazin-l -yl)pyrazoIo[l ,5-a]pyrimidin-3-yl)methylene)imidazolidine- 2,4-dione. LCMS (ES): >95% pure, m/z 545 [M+H]+.
The molecules described in the following table were prepared using chemistries similar to those exemplified in Example 33. All compounds were characterized by LCMS. Table 5D shows the biological activities of the compounds listed in Table 5C. Table 5C.
Figure imgf000241_0001
Figure imgf000241_0002
Example 34. Synthesis of fer/-butyl 5-(3-chloro-5-cvanopyridin-2-ylamino)-3- formylpyrazolo[K5-a1pyrimidin-7-yl(cyclopropyl)carbamate
Figure imgf000242_0001
7¾AY-butyl 5-chloro-3-formylpyrazolo[l ,5-a]pyrimidin-7-yl(cyclopropyl)carbamate (0.5 g, 1 .5 mmol) and 2-amino-3-chloro-5-cyanopyridine (340 mg, 2.3 mmol) were dissolved in anhydrous THF (4.5 mL). Sodium /er/-butoxide (220 mg, 2.3 mmol) was added in one portion. After 1 h, additional 2-amino-3-chloro-5-cyanopyridine (230 mg, 1.5 mmol) and sodium tert- butoxide ( 140 mg, 1.5 mmol) were added. After 4 hours, the reaction was poured into cold H2O (25 mL). The orange precipitate was filtered off and the filtrate was extracted with EtOAc (3 x 50 mL). The organics were washed with brine (1 x 100 mL), dried over MgSC>4, filtered and concentrated in vacuo to provide te -butyl 5-(3-chloro-5-cyanopyridin-2-ylamino)-3- formylpyrazolo[l ,5-a]pyrimidin-7-yl(cyclopropyl)carbamate (670 mg, 99%) as a tan solid. LCMS (ES): >90% pure, m/z 454 [M+H]+.
Example 35. Synthesis of 5-chloro-6-(7-(cvclopropylamino)-3-formylpyrazolo[l .5-a]pyrimidin-
5-ylamino)nicotinonitrile 2,2,2-trifluoroacetate
Figure imgf000242_0002
Tert-buXy\ 5-(2-bromo-4-cyanophenylamino)-3-formylpyrazolo[l ,5-a]pyrimidin-7- yl(cyclopropyl)carbamate (250 mg, 0.55 mmol) was dissolved in dichloromethane (2.5 mL) and trifluoroacetic acid (2.5 mL) was added. After 1 h, the reaction was concentrated under a stream of air and the residue was triturated with Et20 (10 mL). After 30 minutes of stirring, the tan solid was collected and dried to give 5-chIoro-6-(7-(cyclopropylamino)-3-formylpyrazolo[l ,5- a]pyrimidin-5-ylamino)nicotinonitrile 2,2,2-trifluoroacetate (132 mg, 51 %). LCMS (ES): >90% pure, m/z 354 [M+H]+. Example 36. Synthesis of (Z)-5-chloro-6-(7-(cvclopropylaminoV3-((l -methyI-2,5- dioxoimidazolidin-4-ylidene methvnpyrazolof l ,5-a1pyrimidin-5-ylamino)nicotinonitrile
Figure imgf000243_0001
3-Methylimidazolidine-2,4-dione was prepared according to the literature procedure setforth in Eur. JOC 2002, 1763. 5-Chloro-6-(7-(cyclopropylamino)-3-forrnylpyrazolo[l ,5- o]pyrimidin-5-ylamino)nicotinonitrile 2,2,2-trifluoroacetate (38 mg, 0.1 mmol) was suspended in EtOH (1 mL). 3-methylimidazolidine-2,4-dione ( 17 mg, 0.15 mmol) and piperidine (25 iL, 0.25 mmol) were added and the reaction was heated to 80 °C. After 15 h, the solution was filtered while it was still warm and the filter cake was washed with warm EtOH (3 mL) to provide (Z)-5- chloro-6-(7-(cyclopropylamino)-3-((l -methyl-2,5-dioxoimidazolidin-4- ylidene)methyl)pyrazolo[l ,5-a]pyrimidin-5-ylamino)nicotinonitrile (35 mg, 94%) as a bright yellow solid . LCMS (ES): >90% pure, m/z 450 [M+H]+. Example 37. Synthesis of 7-N-cvclopropyl-5-N-(4-morpholinophenv0pyrazolon .5- fllpyrimidine-5 7-diamine hydrochloride
Figure imgf000243_0002
5-Chloro-N-cyclopropylpyrazolo[l ,5-a]pyrimidin-7-amine (20.24 g, 96.9 mmol) was suspended in EtOH (100 mL). 4-Morpholinoaniline (25.93 g, 145 mmol) was added.
Concentrated HCI (9.6 mL, 1 16 mmol) was added and the solution was placed in a 95 °C oil bath. After 72 h, the solution was cooled to 21 °C. The faint purple precipitate was collected and wash with EtOH ( 100 mL). The solid was triturated with EtOAc (200 mL) and further washed with EtOAc (100 mL). Overnight drying in a vacuum oven (30 mmHg, 50 °C) afforded 7-N-cyclopropyl-5-N-(4-morpholinophenyl)pyrazolo[l ,5-a]pyrimidine-5,7-diamine
hydrochloride (25.4 g, 68%) as an off white solid. LCMS (ES): >95% pure, m/z 351 [M+H]+.
Example 38. Synthesis of 7- cvclopropylamino')-5-(4-moφholinophenylamino)pyrazoloΓ l ,5- q]pyrimidine-3-carbaldehvde
Figure imgf000244_0001
7-N-Cyclopropyl-5-N-(4-morpholinophenyl)pyrazolo[ l ,5- ]pyrimidine-5,7-diamine hydrochloride (7.5 g, 19.4 mmol) was suspended in anhydrous DMF (38 mL). The solution was cooled to 0 °C and then POCI3 (2.3 mL, 25.2 mmol) was added dropwise at a rate such that the internal temperature was maintained <5 °C. After POC addition, the solution was dark green and homogeneous. The solution was allowed to warm to 21 °C. After ~1 h, a green precipitate started to form. After 24 h, the solution was added to cold ¾0 (300 mL) and the pH was adjusted to 1 1 by addition of 6Ν NaOH. After stirring for an additional hour, the precipitate was collected by filtration. The solid was partitioned between dichloromethane (1 50 mL) and H20 (150 mL). The aqueous layer was further extracted with dichloromethane (2 x 150 mL). The organics were washed with brine (1 x 500 mL), dried over MgS0 , filtered, and concentrated in vacuo. The solid was triturated with dichloromethane and the filtrate was concentrated in vacuo. The residue was subjected to this protocol two additional times to yield three crops of 7- (c clopropylamino)-5-(4-moφholinophenyta
(3.0 g, 41 %). LCMS (ES): >95% pure, m/z 379 [M+H]+.
Example 39. Synthesis of (Z)-5-((7-(cvclopropylaminoV5-(4- morpholinophenylamino')pyrazolo l ,5-a]pyrimidin-3-ynmethylene)-3-methylimidazolidine-2,4- dione
Figure imgf000245_0001
7-(Cyclopropylamino)-5-(4-moφholinophenylamino)pyrazolo[l ,5-α]pyrimidine-3- carbaldehyde (25 mg, 0.066 mmol) was suspended in EtOH (0.5 mL). 3-Methylimidazolidine- 2,4-dione (12 mg, 0.1 mmol) and piperidine (10 μί, 0.1 mmol) were added and the reaction was heated to 80 °C. After 18 h, the solution was filtered while it was still warm and the filter cake was washed with warm EtOH (3 mL) to give (Z)-5-((7-(cyclopropylamino)-5-(4- morpholinophenylamino)pyrazolo[l ,5-a]pyrimidin-3-yl)methylene)-3-methylimidazolidine-2,4- dione (30 mg, 96%) as a bright yellow solid . LCMS (ES): >95% pure, m/z 475 [M+H]+.
Example 40. Synthesis of 7-N-cvclopropyl-5-N- 2-I^uoro-6-methoxyphenyl pyrazolo[h5- fll yrimidine-5,7-diamine
Figure imgf000245_0002
5-Chloro-N-cyclopropylpyrazolo[l ,5-a]pyrimidin-7-amine (6.5 g, 31 .1 mmol) was suspended in EtOH (33 mL). 2-Fluoro-6-methoxyaniline (8.77 g, 62.2 mmol) was added and the solution became homogeneous. Concentrated HC1 (3.9 mL, 46.7 mmol) was added and the solution was placed in a 95°C oil bath. After 72 h, the volatiles were removed in vacuo. The purple oil was partitioned between dichloromethane (250 mL) and IN NaOH (250 mL) and the organic layer was concentrated in vacuo. The resulting solid was triturated with EtOAc (100 mL) and collected by filtration to afford 7-N-cyclopropyl-5-N-(2-fluoro-6- methoxyphenyl)pyrazolo[l ,5-a]pyrimidine-5,7-diamine (7.82 g, 80%) as a white solid. LCMS (ES): >95% pure, m/z 314 [M+H]+.
Example 41 . Synthesis of 7-(cyclopropylamino>5-(2-fluoro-6- methoxyphenylamino)pyrazolo[l ,5-a]pyrimidine-3-carbaldehyde
Figure imgf000246_0001
7-N-Cyclopropyl-5-N-(2-fluoro-6-methoxyphenyl)pyrazolo[l ,5-a]pyrimidine-5,7- diamine (7.79 g, 24.86 mmol) was suspended in anhydrous DMF (40 mL). The solution was cooled to 0 °C and then POCI3 (3.0 mL, 32.3 mmol) was added dropwise at a rate such that the internal temperature was maintained <5 °C. After POCI3 addition, the solution became progressively more yellow and homogeneous. The solution was allowed to warm to 21 °C. After 24 h, the solution was added to cold H20 (400 mL) and the pH was adjusted to 1 1 by addition of 6Ν NaOH. After stirring for an additional hour, the precipitate was collected by filtration. Trituration of the solid with 30% dichloromethane/hexanes (200 mL), then dichloromethane (100 mL) yielded 7-(cyclopropylamino)-5-(2-fluoro-6- methoxyphenylamino)pyrazolo[l ,5-a]pyrimidine-3-carbaldehyde (3.54 g, 42%) as a white solid. LCMS (ES): >95% pure, m/z 342 [M+H]+. Example 42. Synthesis of (Z)-5-((7-(cyclopropylamino)-5-(2-fluoro-6- methoxyphenylamino)pyrazolori ,5-alpyrimidin-3-vnmethylene")-3-methylimidazolidine-2,4- dione
Figure imgf000247_0001
7-(Cyclopropylamino)-5-(2-fluoro-6-methoxyphenylamino)pyrazolo[l ,5-a]pyrimidine-3- carbaldehyde (75 mg, 0.22 mmol) was suspended in EtOH (2.2 mL). 3-Methylimidazolidine- 2,4-dione (30 mg, 0.26 mmol) and piperidine (26 μΐ^, 0.26 mmol) were added and the reaction was heated to 80 °C. After ~15 min, the reaction became homogeneous. After 18 h, the solution was filtered while it was still warm and the filter cake was washed with warm EtOH (3 mL) to give (Z)-5-((7-(cyclopropylamino)-5-(2-fluoro-6-methoxyphenylamino)pyrazolo[l ,5- fl]pyrimidin-3-yl)methylene)-3-methylimi azolidine-2,4-dione (78 mg, 81 %) as a bright yellow solid . LCMS (ES): >95% pure, m/z 438 [M+H]+.
Example 43. Synthesis of (Z)-4H7-(cyclopropylamino)-3-(n -fhydroxymethvO^.S- dioxoimidazolidin^-ylidene^ethyOpyrazolof 1 ,5-alpyrimidin^^^
Figure imgf000247_0002
Same procedure as Example 1 1. LCMS (ES):>90% pure, m/z 449 [M+H]+. Example 44. Synthesis of (S,ZH4-((5-f4-cvano-2-fluorophenylamino^
('cyclopropylamino')pyrazolori ,5-alpyrimidin-3-yl methylene)-2.5-dioxoimidazolidin- l - yQmethyl 2-("tert-butoxycarbonylamino')propanoate
Figure imgf000248_0001
To (Z)-4-(7-(cyclopropylamino)-3-((l -(hydroxymethyl)-2,5-dioxoimidazolidin-4- ylidene)methyl)pyrazolo[l ,5-a]pyrimidin-5-ylamino)-3-fluorobenzonitrile ( 160 mg, 0.36 mmol) in DMF (5 mL) was added Boc-L-Ala-OH (135 mg, 0.71 mmol), EDCI ( 137 mg, 0.71 mmol), and DMAP (13 mg, 0.1 1 mmol). After stirring at rt for 30 min, additional Boc-L-Ala-OH (135 mg, 0.71 mmol) and EDCI (137 mg, 0.71 mmol) was added and continued to stir at rt for 2h. The reaction mixture was diluted with 3 volumes of water while stirring and the resulting precipitate was filtered off and washed with water. The material was purified by silica gel chromatography eluting with 20%-40% EtOAc/CI- Ch gradient. The pure fractions were combined to provide 213 mg of (S,Z)-(4-((5-(4-cyano-2-fluorophenylamino)-7-(cyclopropylamino)pyrazolo[l ,5- a]pyrimidin-3-yl)methylene)-2,5-dioxoimidazolidin-l -yl)methyl 2-(tert- butoxycarbonylamino)propanoate (LCMS (ES): >95% pure, m/z 620 [M+H]+. Example 45. Synthesis of fS,Z)-f4-((5-(4-cyano-2-fluorophenylamino)-7-
("cvclopropylamino pyrazolo|" 5-alpyrimidin-3-yl)methylene)-2,5-dioxoimidazolidin- l - vOmethyl 2-aminopropanoate hydrogen chloride
Figure imgf000249_0001
To (S,Z)-(4-((5-(4-cyano-2-fluorophenylamino)-7-(cyclopropylamino)pyrazolo[l ,5- a]pyrimidin-3-yl)methylene)-2,5-dioxoirnidazolidin-l -yl)methyl 2-(tert- butoxycarbonylamino)propanoate (213 mg, 0.34 mmol) was added 2M HCl/ether (6 mL) and the reaction mixture was stirred at rt for 2h. The solid was filtered, washed with ether, and dried under vacuum to provide (S,Z)-(4-((5-(4-cyano-2-fluorophenylamino)-7- (cyclopropylamino)pyrazolo[l ,5-a]pyrimidin-3-yl)methylene)-2,5-dioxoimidazolidin- l - yl)methyl 2-aminopropanoate hydrogen chloride. (LCMS (ES): >95% pure, m/z 520 [M+H]+.
Example 46. Synthesis of (ZV5-((5-(3-chloropyridin-4-ylamino')-7-
Ccvclopropylamino')pyrazolo 1.5-alpyrimidin-3-yl')methylene)-3-(hydroxymethyl')imidazolidine- 2,4-dione
Figure imgf000249_0002
Same procedure as Example 1 1 . LCMS (ES):>90% pure, m/z 441 [M+H]+. Example 47. Synthesis of (S,Z)-(4-f(5-(3-chloropyridin-4-ylaminoV7- (cyclopropylamino'lpyrazolori .S-a pyrimidin-S-vnmethylene^^.S-dioxoimidazolidin- l - ynmethyl 2-ftert-butoxycarbonylamino")propanoate
Figure imgf000250_0001
To (Z)-5-((5-(3-chloropyridin-4-ylamino)-7-(cyclopropylamino)pyrazolo[ l ,5- a]pyrimidin-3-yl)methylene)-3-(hydroxymethyl)imidazolidine-2,4-dione (74 mg, 0.17 mmol) in DMF (2 mL) was added Boc-L-Ala-OH (64 mg, 0.34 mmol), EDCI (64 mg, 0.34 mmol), and DMAP (6 mg, 0.05 mmol). After stirring at rt for 30 min, additional Boc-L-Ala-OH (32 mg,
0.17 mmol) and EDCI (32 mg, 0.17 mmol) was added and continued to stir at rt for 30 min. The reaction mixture was diluted with 3 volumes of water while stirring and the resulting precipitate was filtered off and washed with water. The solid was dried under vacuum to provide 100 mg of (S,Z)-(4-((5-(3-chloropyridin-4-ylamino)-7-(cyclopropylamino)pyrazolo[l ,5-a]pyrimidin-3- yl)methylene)-2,5-dioxoimidazolidin-l -yl)methyl 2-(tert-butoxycarbonylamino)propanoate (LCMS (ES): >95% pure, m/z 612 [M+H]+.
Example 48. Synthesis of ("S,Z)-(4-(Y5-i3-chloropyridin-4-ylamino")-7-
(cyclopropylamino)pyrazolori ,5-alpyrimidin-3-yl)methylene)-2,5-dioxoimidazolidin- l - vDmethyl 2-aminopropanoate hydrogen chloride
Figure imgf000251_0001
Same procedure as Example 45. LCMS (ES):>90% pure, m/z 512 [M+H]+.
Table 6 shows the biological activities of certain selected compounds of Exampl and 42 to 48.
Biological Activities of Selected Exemplary Compounds
Figure imgf000251_0003
Example 49. Synthesis of fe/7-butyl 5-(3-chloropyridin-4-ylaminoV3-formylpyrazolo[l ,5- alpyrimidin-7-yl("cyclopropylmethv0carbamate
Figure imgf000251_0002
7er/-butyl 5-chloro-3-formylpyrazolo[l ,5-a]pyrimidin-7- yl(cyclopropylmethyl)carbamate (75 mg, 0.21 mmol) was dissolved in 1 ,4-dioxane ( 1.5 mL). 4- Amino-3-chloropyridine (41 mg, 0.32 mmol), Cs2C03 (98 mg, 0.30 mmol), (±)-BlNAP (9 mg, 0.015 mmol) and palladium(II) acetate (7 mg, 0.01 mmol) were then added. The mixture was sealed and irradiated at 1 10 °C for 20 min in the microwave. Et20 (8 mL) was added and the filtrate was concentrated in vacuo to a yellow/orange foam. The crude residue was purified via flash column chromatography (30-60% EtOAc/hexanes) to yield /er/-butyl 5-(3-chloropyridin-4- ylamino)-3-formylpyrazolo[l ,5-a]pyrimidin-7-yl(cyclopropylmethyl)carbamate (42 mg, 44%) as a yellow solid. LCMS (ES): >90% pure, m/z 443 [M+H]+.
Example 50. Synthesis of 5-(3-chloropyridin-4-ylaminoV7-
(cvclopropylmethylamino)pyrazolo[K5- lpyrimidine-3-carbaldehyde 2,2,2-trifluoroacetate
Figure imgf000252_0001
Tert-buty] 5-(3-chloropyridin-4-ylamino)-3-formylpyrazolo[l ,5-a]pyrimidin-7- yl(cyclopropylmethyl)carbamate (40 mg, 0.09 mmol) was dissolved in dichloromethane (1 mL) and trifluoroacetic acid ( 1 mL). After one hour, the solution was concentrated under a stream of air. The crude residue was triturated with hexanes (4 mL) and filtered to furnish 5-(3- chloropyridin-4-ylamino)-7-(cyclopropylmethylamino)pyrazolo[l ,5-o]pyrimidine-3- carbaldehyde 2,2,2-trifluoroacetate (28 mg, 68%) as an off white solid. LCMS (ES): >95% pure, m/z 343 [M+H]+. Example 51 . Synthesis of ("Z)-5-((5-(3-chloropyridin-4-ylaminoV7-
(cvclopropylmethylamino)pyrazolori ,5-a1pyrimidin-3-yl')methylene')imidazolidine-2,4-dione
Figure imgf000253_0001
5-(3-Chloropyridin-4-ylamino)-7-(cycto
3-carbaldehyde 2,2,2-trifluoroacetate (28 mg, 0.06 mmol) was suspended in EtOH (1 .2 mL). Hydantoin (9 mg, 0.09 mmol) and piperidine (22 μί, 0.23 mmol) were added and the reaction was heated to 80 °C. After 18 h, the solution was filtered while warm and the filter cake was washed with warm EtOH (3 mL) to afford (Z)-5-((5-(3-chloropyridin-4-ylamino)-7- (cyclopropylmethylamino)pyrazolo[l ,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione ( 16 mg, 61 %) as a yellow solid. LC S (ES): >95% pure, m/z 425 [M+H]+.
Table 7 shows the biological activity of Example 51 . That is, Compound 3U in Table 7 is Example 51 .
Table 7.
Figure imgf000253_0002
Example 52. Synthesis of (Z)-5-ff2-fO'-chloro-4-fluorobiphenyl-2-yl)methylamino')-4- (cyclopropylamino)pyrazolon ,5-a][ l .3,51triazin-8-yOmethylene)imidazolidine-2,4-dione
Figure imgf000254_0001
To the reaction flask, (Z)-5-((2-(2-bromo-5-fluorobenzylamino)-4- (cyclopropylamino)pyrazolo[l ,5-a][l )3,5]triazin-8-yl)rnethylene)imidazolidine-2,4-dione (10 mg, 0.02 mmol) was suspended in DMF. To the mixture was added NaOAc (8.4 mg, 0.1 mmol) and 3-chlorophenylboronic acid (4.8 mg, 0.03 mmol). The mixture was degassed under nitrogen for 3 minutes, then PdC^dppf (0.8 mg, <0.01 mmol) was added. The reaction mixture was then heated in microwave for 10 minutes at 120°C, diluted with NMP, and prepared by HPLC purification to provide (Z)-5-((2-((3'-chloro-4-fluorobiphenyl-2-yl)methylamino)-4- (cyclopropylamino)pyrazolo[l ,5-a][l,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione as a solid (2.1 mg). LCMS (ES):>95% pure, m/z 519 [M+H]+.
The compounds in the following Table 7A were prepared using procedures described in Example 52 by using the appropriate starting boronic acids. Compounds were prepared by HPLC purification and concentrated down in Genevac. Some required further TLC purification (2% MeOH in DCM). Table 7B shows the the biological activities of the compounds listed in Table 7A. That is, Compounds 3V to 41 in Table 7B are the compounds listed in Table 7A.
Table 7A.
Figure imgf000254_0002
Figure imgf000255_0001
Figure imgf000256_0001
Table 7B.
Figure imgf000256_0002
Figure imgf000257_0001
Example 53. Synthesis of (ZV5-((2-(2-bromobenzylamino)-4-(cvclopropylamino pyr*azolo[l ,5- airi 3,51triazin-8-y0methylene)imidazolidine-2,4 lione
Figure imgf000257_0002
To the reaction flask, a mixture of (1 : 1 ) (Z)-5-((4-(cyclopropylamino)-2- (methylsulfinyl)pyrazolo[l ,5-a][l ,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione and (Z)-5- ((4-(cyclopropylamino)-2-(methylsulfonyl)pyrazolo[l ,5-a][l ,3,5]triazin-8- yl)methylene)imidazolidine-2,4-dione (500 mg, 1 .4 mmol) was suspended in isopropanol. To the mixture was added DIEA (450 iL, 2.8 mmol) and (2-bromophenyl)methanamine HC1 (610 mg, 2.8 mmol). The mixture was stirred at 90°C for five and a half hours. The mixture was cooled to room temperature and resulting precipitate was filtered and washed with isopropanol to provide (Z)-5-((2-(2-bromobenzylamino)-4-(cyclopropylamino)pyrazolo[l ,5-a][l ,3,5]triazin-8- yl)methylene)imidazolidine-2,4-dione as a solid (610 mg). LCMS (ES):>95% pure, m z 471 [M+2H]+. Example 54. Synthesis of (Z)-5-f(2-(Y3'-chlorobiphenyl-2-yl)methylamino)-4- (cyclopropylamino'lpyrazolor S-airU^ltriazin-S-vnmethylene midazolidine^^-dione
Figure imgf000258_0001
To the reaction flask, (Z)-5-((2-(2-bromobenzylamino)-4- (cyclopropylamino)pyrazolo[l,5-a][l ,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione (10 mg, 0.02 mmol) was suspended in DMF. To the mixture was added NaOAc (8.7 mg, 0.1 mmol) and 3-chlorophenylboronic acid (5 mg, 0.03 mmol). The mixture was degassed under nitrogen for three minutes, then PdC^dppf (0.8 mg, <0.01 mmol) was added. The mixture was heated in microwave at 120°C for 10 minutes. The mixture was diluted with NMP and subjected to preparative HPLC purification to provide (Z)-5-((2-((3'-chlorobiphenyl-2-yl)methylamino)-4- (cyclopropylamino)pyrazolo[l,5-a][l ,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione as a solid (1.5 mg). LCMS (ES):>95% pure, m/z 501 [M+H]+
The compounds in the following Table 8 A were prepared using procedures described in
Example 54 by using the appropriate starting boronic acids. Compounds were prepared by HPLC purification and concentrated down in Genevac. Some required further TLC purification (2% MeOH in DCM). Table 8B shows the biological activities of the compounds listed in Table 8A and Example 54.
Table 8A.
Figure imgf000258_0002
Figure imgf000259_0001
Figure imgf000260_0001
Table 8B.
Figure imgf000260_0002
Figure imgf000261_0001
Example 55. Synthesis of ("ZV5-C('2-('2-bromo-4-fluorobenzylamino')-4- ('cvclopropylamino')pyrazolo[K5-al[l ,3,51triazin-8-yl')methylene')imidazolidine-2,4-dione
Figure imgf000261_0002
To the reaction flask, a mixture of (1 : 1 ) (Z)-5-((4-(cyclopropylamino)-2- (methylsulfinyl)pyrazolo[l ,5-a][l ,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione and (Z)-5- ((4-(cyclopropylamino)-2-(methylsulfonyl)pyrazolo[l ,5-a][l ,3,5]triazin-8- yl)methylene)imidazolidine-2,4-dione (500 mg, 1 .4 mmol) was suspended in isopropanol. To the mixture was added DIEA (450 μί, 2.8 mmol) and (2-bromo-4-fluorophenyl)methanamine (660 mg, 2.8 mmol). The mixture was stirred at 90°C for two days. The mixture was then let to cool to room temperature and solid filtered, then washed with isopropanol to provide (Z)-5-((2- (2-bromo-4-fluorobenzylamino)-4-(cyclopropylamino)pyrazolo[l ,5-a][l ,3,5]triazin-8- yl)methylene)imidazolidine-2,4-dione as a solid (610 mg). LCMS (ES):>95% pure, m/z 487 [M+H]+.
Example 56. Synthesis of (Z)-5-((2-((3'-chloro-5-fluorobiphenyl-2-vnmethylamino)-4- (cyclopropylamino^pyrazolorKS-alf U^ltriazin-S-vDmethylene midazolidine^^-dione
Figure imgf000262_0001
To the reaction flask, (Z)-5-((2-(2-bromo-4-fluorobenzylamino)-4- (cyclopropylamino)pyrazolo[l ,5-a][l,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione (10 mg, 0.02 mmol) was suspended in DMF. To the mixture was added NaOAc (8.4 mg, 0.1 mmol) and 3-chlorophenylboronic acid (4.8 mg, 0.03 mmol). The mixture was degassed under nitrogen for 3 minutes, then PdC dppf (0.8 mg, <0.01 mmol) was added. The reaction mixture was then heated in microwave for 10 minutes at 120°C, diluted with NMP, and prepared by HPLC purification to provide (Z)-5-((2-((3'-chloro-5-fluorobiphenyl-2-yl)methylamino)-4- (cyclopropylamino)pyrazolo[l ,5-a][l ,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione as a solid (3.8 mg). LCMS (ES):>95% pure, m/z 519 [M+H]+.
The compounds in the following Table 9A were prepared using procedures described in Example 56 by using the appropriate starting boronic acids. Compounds were prepared by HPLC purification and concentrated down in Genevac. Some required further TLC purification (2% MeOH in DCM). Table 9B shows the biological activities of the compounds listed in Table 9A.
Table 9A.
Figure imgf000262_0002
Figure imgf000263_0001
Figure imgf000264_0001
Table 9B.
Figure imgf000264_0002
Example 57. Synthesis of (ZV5-rC4-(cvclopropylamino')-2-('('5-ethylpyridin-2- yl')methoxy pyrazolon .5-ain .3,51triazin-8-yl')methylene')imidazolidine-2.4-dione
Figure imgf000265_0001
To the reaction flask, (5-ethylpyridin-2-yl)methanol ( 19 mg, 0.1 mmol) and sodium hydride (5.5 mg, 0.1 mmol of 60%wt) was suspended in NMP. The mixture was stirred at room temperature for seventy five minutes. A mixture of (1 : 1 ) (Z)-5-((4-(cyclopropylamino)-2-
(methylsulfinyl)pyrazolo[l ,5-a][l ,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione and (Z)-5- ((4-(cyclopropylamino)-2-(methylsulfonyl)pyrazolo[l ,5-a][l ,3,5]triazin-8- yl)methylene)imidazolidine-2,4-dione (10 mg, 0.03 mmol) was added and the mixture was stirred at room temperature for sixteen hours. The mixture was diluted with water and NMP then subjected to preparative HPLC purification. Genevac evaporation provided (Z)-5-((4- (cyclopropylamino)-2-((5-ethylpyridin-2-yl)methoxy)pyrazolo[l ,5-a][l ,3,5]triazin-8- yl)methylene)imidazolidine-2,4-dione as a solid (12 mg). LCMS (ES): 100% pure, m/z 421 [M+H]+.
The compounds in the following table were prepared using procedures described in Example 57 by using the appropriate starting alcohols. Compounds were prepared by HPLC purification and concentrated down in Genevac. Table 10B shows the biological activities of the compounds listed in Table 10A and Example 57. That is, Compounds 5M to 5P are the compounds listed in Table 10A and Example 57. Table 10A.
Figure imgf000265_0002
Figure imgf000266_0001
Table 10B.
Figure imgf000266_0003
Example 58. Synthesis of (ZVS-W-fcvclopropylamino'lpyrazolo l .S-airU.Sltnazin-S- yDmethylene)imidazolidine-2,4-dione
Figure imgf000266_0002
To a mixture of (Z)-5-((4-(cyclopropylamino)-2-(methylsulfinyl)pyrazolo[l,5- a][l ,3,5]triazin-8-yl)methyIene)imidazolidine-2,4-dione and (Z)-5-((4-(cyclopropylamino)-2- (methylsulfonyl)pyrazolo[l ,5-a][l ,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione (25 mg, 0.07 mmol) in isopropanol (2 mL) was added sodium borohydride (6 mg, 0.17 mmol). The reaction mixture was stirred at rt for 1 hour, filtered through a PTFE filter, and purified by mass- directed LC/MS to provide (Z)-5-((4-(cyclopropylamino)pyrazolo[l ,5-a][l ,3,5]triazin-8- yl)methylene)imidazolidine-2,4-dione as the TFA salt. (LCMS (ES): >95% pure, m/z 466
[M+H]+.
Table 1 1. Biological Activity of Example
Figure imgf000267_0002
Example 59. Synthesis of (Z)-5-((2-(5-chloro-2-methylphenylamino)-4- (cvclopropylamino)pyrazolori ,5-airi ,3.51triazin-8-vnmethylene)imidazolidine-2.4-dione
Figure imgf000267_0001
To the reaction flask, a mixture of (1 : 1) (Z)-5-((4-(cyclopropylamino)-2- (methylsulfinyl)pyrazolo[l ,5-a][l ,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione and (Z)-5- ((4-(cyclopropylamino)-2-(methylsulfonyl)pyrazolo[l ,5-a][l ,3,5]triazin-8- yl)methylene)imidazolidine-2,4-dione (10 mg, 0.03 mmol) was suspended in isopropanol. To the mixture was added PTSA (7 mg, 0.04 mmol) and 5-chloro-2-methylaniline (39 mg, 0.3 mmol). The mixture was stirred at 90°C for seven days. The mixture was diluted with NMP then subjected to preparative HPLC purification. Genevac evaporation provided (Z)-5-((2-(5-chloro- 2-methylphenylamino)-4-(cyclopropylamino)pyrazolo[l ,5-a][l ,3,5]triazin-8- yl)methylene)imidazolidine-2,4-dione as a solid (1.7 mg). LCMS (ES):>90% pure, m/z 425 [M+H]+.
The compounds in the following Table 1 1 A were prepared using procedures described in Example 59 by using the appropriate starting boronic acids. Compounds were prepared by HPLC purification and concentrated down in Genevac. Table 1 IB shows the biological activities of the compounds listed in Table 1 1 A as well as Examples 59 to 62. That is, Compounds 5R to 5X in Table 1 I B are the compounds listed in Table 1 1 A and Examples 59 to 62. Table 1 1 A.
Figure imgf000268_0001
Example 60. Synthesis of (ZVmethyl 2-(4-r4-(cvclopropylaminoV8-((2,5-dioxoimidazolidin-4- ylidene')methyl')pyrazolo 1.5-airi ,3,5]triazin-2-ylamino')phenoxy)acetate
Figure imgf000269_0001
To a mixture of (Z)-5-((4-(cyclopropylamino)-2-(methylsulfinyl)pyrazolo[l ,5- a][l ,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione and (Z)-5-((4-(cyclopropylamino)-2- (methylsulfonyl)pyrazolo[l ,5-a][l ,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione (25 mg, 0.07 mmol) in isopropanol (2 mL) was added -toluenesulfonic acid (13 mg, 0.07 mmol) and methyl 2-(4-aminophenoxy)acetate (62 mg, 0.35 mmol). The reaction mixture was stirred under microwave heating at 1 10°C for 20 min. The reaction mixture was cooled to 0°C and the resulting solid was filtered off, washed with isopropanol and dried under vacuum to provide (Z)- methyl 2-(4-(4-(cyclopropylamino)-8-((2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[l ,5- a][l ,3,5]triazin-2-ylamino)phenoxy)acetate. (LCMS (ES): >95% pure, m/z 465 [M+H]+.
Example 61 . Synthesis of (Z)-ethyl 6-(4-(4-(cvclopropylamino)-8-((2,5-dioxoimidazolidin-4- ylidene)methyl")pyrazolo[l ,5-al|" 1.3.51triazin-2-ylamino)phenoxy)hexanoate
Figure imgf000269_0002
Same procedure as Example 60. LCMS (ES):>90% pure, m/z 535 [M+H]+. Example 62. Synthesis of (Z)-2-("4-(4-(cyclopropylaminoV8-((2,5-dioxoimidazolidin-4-
Figure imgf000269_0003
Figure imgf000270_0001
To (Z)-methyl 2-(4-(4-(cyclopropylamino)-8-((2,5-dioxoimidazolidin-4- ylidene)rnethyl)pyrazolo[l ,5-a][l ,3,5]triazin-2-ylarnino)phenoxy)acetate (20 mg, 0.04 mmol) in methanol (1 mL) was added hydroxy lamine ((2 mL of the supernatent) which was previously prepared by adding powdered KOH (320 mg, 5.70 mmol) to hydroxylamine hydrogen chloride (250 mg, 3.62 mmol) in methanol (3 mL) and stirring at rt for 30 min). The reaction mixture was stirred at rt for 30 min. The crude material was filtered through a PTFE filter and purified by mass-directed LC/MS to provide (Z)-2-(4-(4-(cyclopropylamino)-8-((2,5-dioxoimidazolidin- 4-ylidene)methyl)pyrazolo[l ,5-a][l ,3,5]triazin-2-ylamino)phenoxy)-N-hydroxyacetamide as the TFA salt. (LCMS (ES): >95% pure, mix 466 [M+H]+.
Table 1 1 B.
Figure imgf000270_0002
Example 63. Synthesis of N-fnr^^-^-rcvclopro ylamino S-frZ t' .S-dioxoimidazolidin^- ylidene')methyl')pyrazolori ,5-a ri .3.51triazin-2-ylamino')cvclohexyl')-4- fluorobenzenesulfonamide
Figure imgf000271_0001
To the reaction flask, a mixture of (1 : 1 ) (Z)-5-((4-(cyclopropylamino)-2- (methylsulfinyl)pyrazolo[l ,5-a][l ,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione and (Z)-5- ((4-(cyclopropylamino)-2-(methylsulfonyl)pyrazolo[l ,5-a][l ,3,5]triazin-8- yl)methylene)imidazolidine-2,4-dione (10 mg, 0.03 mmol) was suspended in isopropanol. N- ((lr,4r)-4-aminocyclohexyl)-4-fluorobenzenesulfonamide (22 mg, 0.1 mmol) and DIEA (14 μΐ,, 0.1 mmol) were added and the mixture was heated at 90°C for seven days. The mixture was diluted with NMP and subjected to preparative HPLC purification to yield N-((l r,4r)-4-(4- (cyclopropylamino)-8-((Z)-(2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[l ,5- a][l ,3,5]triazin-2-ylamino)cyclohexyl)-4-fluorobenzenesulfonamide as a solid (2 mg). LCMS (ES):>90% pure, mix 556 [M+H]+.
Example 64. Synthesis of 4-cvano-N-(4-{4-cyclopropylamino-8- 2,5-dioxo-imidazolidin-(4Z)- ylidenemethyll-pyrazolo K5-al l ,3,5]triazin-2-ylamino)-cyclohexyO-benzenesulfonamide
Figure imgf000271_0002
Same procedure as Example 64. LCMS (M+ 1 =563) Table 12. Biological Activities of Examples 63 and 64.
Figure imgf000272_0003
Example 65. Synthesis of 5-chloro-N. N-dimethylpyrazolo[h5-q]pyrimidin-7-amine
Figure imgf000272_0001
5,7-Dichloropyrazolo[l ,5-a]pyrimidine (5 g, 26.6 mmol) was suspended in 2-propanol (65 mL). Triethylamine (9.3 mL, 66.5 mmol) was added and then dimethylamine hydrochloride (2.6 g, 31.9 mmol). After the amine addition, a slow exotherm of ~10 °C occurred over 10 minutes. After 2 h, the reaction was poured into water (200 mL) and stirred for 0.5 h. The precipitate was filtered off and washed liberally with water (200 mL). The filter cake was dried overnight in a vacuum oven (50 °C, 30 mmHg) to provide 5-chloro-N, N-dimethylpyrazolo[l ,5- a]pyrimidin-7-amine (3.83 g, 74%) as a white solid. Ή NMR (CDC13, 400 MHz) δ: 7.99 (d, lH, J= 2.4 Hz), 6.44 (d, 1 H, J = 2.4 Hz), 5.94 (s, 1H), 3.40 (s, 6H). LCMS (ES): >95% pure, m/z 197 [M+H]+.
Example 66. Synthesis of N. N-dimethyl-5-("5-methylthiophen-2-ynpyrazolo[1.5-olpyrimidin-7- amine
Figure imgf000272_0002
In a reaction flask, 5-chloro-N, N-dimethylpyrazolo[l ,5-cr]pyrimidin-7-amine (200 mg, 1.02 mmol) and 5-methylthiophene-2-boronic acid (188 mg, 1.33 mmol) were dissolved in toluene (10 mL) under an atmosphere of nitrogen. Ethanol (2 mL), 2M Na2CC>3 (1.5 mL), and PdC dppf CHaC (83 mg, 0.1 mmol) were sequentially added to the starting material. The reaction was placed in an 80°C oil bath. Additional 5-methylthiophene-2-boronic acid (100 mg) and PdCl2dppf CH2Cl2 (50 mg) were added after 1 h. After 22 h, the solution was diluted with brine (25 mL) and extracted with EtOAc (2 x 25 mL). The organics were dried over MgSC>4, filtered, and concentrated in vacuo. The dark brown residue was purified via preparative TLC (20% EtOAc/hexanes) to provide N, N-dimethyl-5-(5-methylthiophen-2-yl)pyrazolo[l ,5- a]pyrimidin-7-amine (66 mg, 25%) as a tan solid. Ή NMR (CDC13, 400 MHz) δ: 8.00 (d, 1H, J = 2.4 Hz), 7.46 (d, 1H, J= 3.6 Hz), 6.79 (d, 1H, J= 3.6 Hz), 6.51 (d, 1 H, J= 2.4 Hz), 6.30 (s, 1 H), 3.36 (s, 6H), 2.54 (s, 3H). LCMS (ES): >75% pure, m/z 259 [M+H]+.
Example 67. Synthesis of 7-(dimethylamino)-5-(5-methylthiophen-2-yl)pyrazolorK5- alpyrimidine-3-carbaldehyde
Figure imgf000273_0001
N, N-Dimethyl-5-(5-methylthiophen-2-yl)pyrazolo[l ,5-a]pyrimidin-7-amine (66 mg, 0.26 mmol) was dissolved in anhydrous DMF (3.3 mL) and the solution was cooled to 0 °C. POC (30 μί, 0.33 mmol) was added dropwise at a rate such that the internal temperature was maintained <5 °C. The solution immediately turned yellow and after the addition a precipitate formed. The reaction was gradually warmed to 21 °C by removal of the ice bath. The solution became homogeneous after <4 h. After 22 h, the solution was added to ice water (30 mL), and 6Ν NaOH was added to pH 1 1. After 1 h of stirring, the solution was filtered and the filter cake was triturated with MeOH (2 x 5 mL) to provide 7-(dimethylamino)-5-(5-methylthiophen-2- yl)pyrazolo[l ,5-a]pyrimidine-3-carbaldehyde (21 mg, 27%) as an off-white solid. "H MR (DMSO-D6, 400 MHz) δ: 10.03 (s, 1 H), 8.47 (s, 1H), 7.93 (d, 1H, J = 3.6 Hz), 6.94 (dd, 1 H, J = 3.6, 1.0 Hz), 6.83 (s, 1 H), 3.42 (s, 6H), 2.52 (s, 3H). LCMS (ES): >95% pure, m/z 287 [M+H]4
Example 68. Synthesis of (Z)-5-((7-(dimethylamino)-5-(5-methylthiophen-2-yl)pyrazolori ,5- fl1pyrimidin-3-yDmethylene)irnidazolidine-2,4-dione
Figure imgf000274_0001
7-(Dimethylamino)-5-(5-methylthiophen-2-yl)pyrazolo[l ,5-a]pyrimidine-3-carbaldehyde (13 mg, 0.045 mmol) was suspended in EtOH (0.5 mL). Hydantoin (7 mg, 0.07 mmol) and piperidine (15 μί, 0.07 mmol) were added and the reaction was heated to 80 °C. After 18 h, the solution was filtered while warm and the filter cake was washed with warm EtOH (3 mL) to afford (Z)-5-((7-(dimethylamino)-5-(5-methylthiophen-2-yl)pyrazolo[l ,5-a]pyrimidin-3- yl)methylene)imidazolidine-2,4-dione (13 mg, 81%) as a bright yellow solid. Ή NMR (DMSO- D6, 400 MHz) δ: 1 1.04 (bs, 1H), 10.82 (bs, 1 H), 8.43 (s, 1H), 7.87 (d, 1H, J = 3.6 Hz), 6.95 (d, 1H, J= 3.6 Hz), 6.67 (s, 1H), 6.54 (s, 1H), 3.40 (s, 6H), 2.52 (s, 3H). LCMS (ES): >95% pure, m/z 369 [M+H]+.
Example 69. Synthesis of 5-chloro-N-methylpyrazolo l ,5-a"|pyrimidin-7-amine
Figure imgf000274_0002
5,7-Dichloropyrazolo[l ,5-a]pyrimidine (5 g, 26.6 mmol) was suspended in 2-propanol (65 mL). Triethylamine (9.3 mL, 66.5 mmol) was added and then methylamine hydrochloride (2.2 g, 31.9 mmol). After the amine addition, a slow exotherm of ~3 °C occurred over 10 minutes. After 2 h, the reaction was poured into water (200 mL) and stirred for 0.5 h. The precipitate was filtered off and washed liberally with water (200 mL). The filter cake was dried overnight in a vacuum oven (50 °C, 30 mmHg) to provide 5-chloro-N-methylpyrazolo[l ,5- a]pyrimidin-7-amine (3.6 g, 74%) as a white solid. Ή NMR (CDC , 400 MHz) δ: 7.97 (d, 1 H, J= 2.0 Hz), 6.49 (bs, 1 H), 6.43 (d, 1 H, J= 2.0 Hz), 5.94 (s, 1 H), 3.12 (d, 3H, J= 5.2 Hz). LCMS (ES): >95% pure, m/z 183 [M+H]+.
Example 70. Synthesis of tert-Buty] 5-chloropyrazolo[L5-a1pyrimidin-7-yl(methyl)carbamate
Figure imgf000275_0001
5-Chloro-N-methylpyrazolo[l ,5-a]pyrimidin-7-amine (1.09 g, 6 mmol) was dissolved in dichloromethane. Di-tert-butyl dicarbonate (1.70 g, 7.8 mmol), triethylamine (1.1 mL, 7.8 mmol), and DMAP (37 mg, 0.3 mmol) were added sequentially. Vigorous bubbling was observed within 5 minutes, After 15 h, the solution was diluted with dichloromethane (20 mL) and water (30 mL). The aqueous layer was further extracted with dichloromethane (2 x 20 mL). The organics were washed with 0.5 N HC1 (1 x 50 mL), then dried over MgS04, filtered, and concentrated in vacuo. The residue was purified via filtration over a plug of silica (20% EtOAc/hexanes), and concentration of the filtrate in vacuo to furnish te -butyl 5- chloropyrazolo[l ,5-a]pyrimidin-7-yl(methyl)carbamate (605 mg, 36%) as a pale yellow solid. Ή NMR (CDCb, 400 MHz) δ: 8.13 (d, 1 H, J= 2.0 Hz), 6.76 (s, 1 H), 6.67 (d, 1 H, J= 2.0 Hz), 3.40 (s, 3H), 1.42 (s, 9H). LCMS (ES): >95% pure, m/z 183 [M+H-Boc]+. Example 71. Synthesis of fe -Butyl methyl(5-(5-methylthiophen-2-vDpyrazolo[l ,5- d\ py ri m i d i n-7-v Dcarbamate
Figure imgf000276_0001
7er/-butyl 5-chloropyrazolo[l ,5-a]pyrimidin-7-yl(methyl)carbamate (300 mg, 1.06 mmol) and 5-methylthiophene-2-boronic acid (227 mg, 1.60 mmol) were dissolved in DME (5 mL) under an atmosphere of nitrogen. Ethanol (2.5 mL), 2M a2CC>3 (1.6 mL), and
PdC dppf-ChhC (43 mg, 0.05 mmol) were added sequentially. The reaction was placed in an 85 °C oil bath. After 6 h, the solution was diluted with brine (50 mL) and extracted with EtOAc (3 x 50 mL). The organics were dried over MgSC>4, filtered, and concentrated in vacuo. The dark brown residue was purified via flash column chromatography (5-10% EtOAc/hexanes) to provide tert-Butyl methyl(5-(5-methylthiophen-2-yl)pyrazolo[l ,5-a]pyrimidin-7-yl)carbamate (136 mg, 37%) as a foamy oil which gradually solidified under high vacuum to a pale yellow solid. Ή NMR (CDCI3, 400 MHz) δ: 8.07 (d, 1 H, J = 2.4 Hz), 7.48 (d, 1 H, J = 3.6 Hz), 7.02 (s, 1 H), 6.82 (dd, 1H, J= 3.6, 1.0 Hz), 6.66 (d, 1 H, J= 2.4 Hz), 3.41 (s, 3H), 2.56 (d, 3H, J= 1.0 Hz ), 1.40 (s, 9H). LCMS (ES): >95% pure, m/z 245 [M+H-Boc]+.
Example 72. Synthesis of /erf-Butyl 3-formyl-5-(5-methylthiophen-2-yl)pyrazolo|"l ,5- fllpyrimidin-7-yl(methyl)carbamate
Figure imgf000276_0002
Terf-butyl methyl(5-(5-methylthiophen-2-yl)pyrazolo[l ,5-a]pyrimidin-7-yl)carbamate (95 mg, 0.28 mmol) was dissolved in anhydrous DMF (1.5 mL) and the solution was cooled to 0 °C. POCI3 (76 μL·, 0.83 mmol) was added dropwise at a rate such that the internal temperature was maintained <5 °C. The reaction was warmed to 21 °C by removal of the ice bath. After 20 h, the solution was added to ice water (25 mL), and the pH was adjusted to 1 1 by addition of 6N NaOH. The precipitate was collected by filtration after 1 h of stirring. The filter cake was dried under high vacuum to yield terf-butyl 3-formyl-5-(5-methylthiophen-2-yl)pyrazolo[l ,5- a]pyrimidin-7-yl(methyl)carbamate (84 mg, 82%) as a pale yellow solid. Ή NMR (CDC13, 400 MHz) δ: 10.31 (s, 1 H), 8.53 (s, 1 H), 7.62 (d, lH, J= 3.6 Hz), 7.18 (s, 1 H), 6.87 (dd, 1H, J= 3.6, 1.0 Hz), 3.43 (s, 3H), 2.59 (s, 3H), 1.41 (s, 9H). LCMS (ES): >95% pure, m/z 273 [M+H-Boc]+.
Example 73. Synthesis of 7-("methylamino)-5-('5-methylthiophen-2-vnpyrazolori .5- a]pyrimidine-3-carbaldehyde
Figure imgf000277_0001
7¾ -butyl 3-formyl-5-(5-methylthiophen-2-yl)pyrazolo[l ,5-o]pyrimidin-7- yl(methyl)carbamate (69 mg, 0.19 mmol) was dissolved in dichloromethane (1 mL).
Trifluoroacetic acid (1 mL) was added and the solution changed color from yellow to orange. After 1 h, the solution was concentrated under a stream of air. Trituration with Et20 (5 mL) gave 7-(methylamino)-5-(5-methylthiophen-2-yl)pyrazolo[l ,5-a]pyrimidine-3-carbaldehyde (44 mg, 88%) as an off white solid. Ή NMR (DMSO-D6, 400 MHz) δ: 10.01 (s, 1H), 8.50 (s, 1H), 8.41 (q, l H, J= 4.8 Hz), 7.91 (d, l H, J= 3.6 Hz), 6.94 (d, l H, J= 3.6 Hz), 6.83 (s, 1H), 3.07 (d, 3H, J = 4.8 Hz), 2.52 (s, 3H). LCMS (ES): >95% pure, m/z 273 [M+H]+.
Example 74. Synthesis of (Z)-5-((7-(methylaminoV5-('5-methylthiophen-2-ynpyrazolo['l ,5- fllpyrimidin-3-vQmethylene)imidazolidine-2,4-dione
Figure imgf000277_0002
7-(Methylamino)-5-(5-methylthiophen-2-yl)pyrazolo[l,5-«]pyri^
(30 mg, 0.1 1 mmol) was suspended in EtOH (1.1 mL). Hydantoin (16 mg, 0.16 mmol) and piperidine (16 ί, 0.16 mmol) were added and the reaction was heated to 80 °C. After 22 h, the solution was filtered while warm and the filter cake was washed with warm EtOH (3 mL) to afford compound (Z)-5-((7-(methylamino)-5-(5-methylthiophen-2-yl)pyrazolo[l ,5-a]pyrimidin- 3-yl)methylene)imidazolidine-2,4-dione (28 mg, 72%) as a bright yellow solid. Ή NMR (DMSO-D6, 400 MHz) δ: 1 1.01 (bs, 2H), 8.42 (s, l H), 8.37 (q, l H, J= 4.8 Hz), 7.87 (d, 1 H, J = 3.6 Hz), 6.96 (dd, 1 H, 7= 3.6, 1.0 Hz), 6.69 (s, 1 H), 6.55 (s, 1 H), 3.07 (d, 3H, J= 4.8 Hz ), 2.52 (s, 3H). LCMS (ES): >90% pure, m z 355 [M+H]+.
Example 75. Synthesis of 5-chloro-N-isopropylpyrazolo[l ,5-alpyrimidin-7-amine
Figure imgf000278_0001
5,7-Dichloropyrazolo[l ,5-a]pyrimidine (5 g, 26.6 mmol) was suspended in 2-propanol (65 mL). Triethylamine (9.3 mL, 66.5 mmol) was added and then isopropylamine (2.74 mL, 31.9 mmol). After the amine addition, a slow exotherm of ~10 °C occurred over 10 minutes. After 15 h, the reaction was poured into water (200 mL) and stirred for 0.5 h. The precipitate was filtered off and washed liberally with water (200 mL). The filter cake was dried overnight in a vacuum oven (50 °C, 30 mmHg) to provide 5-chloro-N-isopropylpyrazolo[l ,5-a]pyrimidin-7- amine (4.17 g, 75%) as a white solid. Ή NMR (CDC13, 400 MHz) δ: 7.96 (d, 1 H, J= 2.4 Hz), 6.42 (d, 1H, J= 2.4 Hz), 6.24-6.38 (m, 1 H), 5.95 (s, 1 H), 3.79-3.92 (m, 1 H), 1 .41 (s, 3H), 1.40 (s, 3H). LCMS (ES): >95% pure, m/z 21 1 [M+H]+.
Example 76. Synthesis of ½r/-butyl S-chloropyrazolon .S-alpyrimidin^-yKisopropyl'toarbamate
Figure imgf000279_0001
5-Chloro-N-isopropylpyrazolo[l ,5-o]pyrimidin-7-amine ( 1.26 g, 6 mmol) was dissolved in dichloromethane (10 mL). Di-/er/-butyl dicarbonate (1.70 g, 12.4 mmol), triethylamine (1 .7 mL, 12.4 mmol), and DMAP (37 mg, 0.3 mmol) were added sequentially. The reaction was placed in a 55°C oil bath. After 24 h, the solution was diluted with dichloromethane (20 mL) and water (30 mL). The aqueous layer was further extracted with dichloromethane (2 x 20 mL). The organics were washed with 0.5 Ν HC1 (1 x 50 mL), then dried over MgS0 , filtered, and concentrated in vacuo. The residue was purified via flash column chromatography (10% EtOAc hexanes) to provide tert-buty\ 5-chloropyrazolo[l ,5- ]pyrimidin-7- yl(isopropyl)carbamate [558 mg, 30% (67% BRSM)] as a white solid. 'H NMR (CDCI3, 400 MHz) δ: 8.1 1 (d, 1 H, 7= 2.4 Hz), 6.66 (s, 1 H), 6.65 (d, 1 H, J = 2.4 Hz), 4.64 (sep, 1 H, J = 6.8 Hz), 1 .28 (s, 9H), 1 .26 (s, 3H), 1 .24 (s, 3H). LCMS (ES): >95% pure, m/z 21 1 [M+H-Boc]+. Example 77. Synthesis of fe -butyl isopropyl("5-f5-methylthiophen-2-yOpyrazolo|T ,5- fllpyrimidin-7-vQcarbamate
Figure imgf000279_0002
7er/-butyl 5-chloropyrazolo[l ,5-a]pyrimidin-7-yl(isopropyl)carbamate (310 mg, 1 mmol) and 5-methylthiophene-2-boronic acid (213 mg, 1 .50 mmol) were dissolved in DME (5 mL) under an atmosphere of nitrogen. Ethanol (2.5 mL), 2M a2C03 (1.5 mL), and
PdChdppf CHaCb (41 mg, 0.05 mmol) were added sequentially. .The reaction was placed in an
85 °C oil bath. After 18 h, the solution was diluted with brine (50 mL) and extracted with EtOAc (3 x 50 mL). The organics were dried over MgSO,), filter*^ and concentrated in vacuo. The dark brown residue was purified via flash column chromatography (5-10% EtOAc/hexanes) to provide tert-buty\ isopropyl(5-(5-methylthiophen-2-yl)pyrazolo[l ,5- ]pyrimidin-7-yl)carbamate (93 mg, 25%) as a yellow solid. Ή NMR (CDC13, 400 MHz) δ: 8.05 (d, 1H, J= 2.4 Hz), 7.49 (d, lH,J=3.6Hz), 6.92 (s, 1H), 6.82 (dd, 1H,J=3.6, 1.0 Hz), 6.64 (d, 1H,J=2.4 Hz), 4.68 (sep, 1H, J =6.8 Hz), 2.56 (d, 3H,J= 1.0 Hz), 1.29 (s,9H), 1.26 (s,3H), 1.25 (s,3H). LCMS (ES): >95% pure, m/z 372 [M+H]+.
Example 78. Synthesis of fe -butyl 3-foimyl-5-f5-methylthiophen-2-vOpyrazolo|"l .5- alpyrimidin-7-yl('isopropyl')carbamate
Figure imgf000280_0001
Terr-butyl isopropyl(5-(5-methylthiophen-2-yl)pyrazolo[l,5-c]pyrimidin-7-yl)carbamate (90 mg, 0.24 mmol) was dissolved in anhydrous DMF (0.5 mL) and the solution was cooled to 0 °C. POCI3 (67 μΐ^, 0.73 mmol) was added dropwise keeping the internal temperature <5 °C.
The reaction was gradually warmed to 21 °C by removal of the ice bath. After 22 h, the solution was added to ice water (20 mL), and 6N NaOH was added to pH 11. After 1 h of stirring the solution was filtered. The filter cake was purified via flash column chromatography (10-15% EtOAc/hexanes) to yield /er/-butyl 3-formyl-5-(5-methylthiophen-2-yl)pyrazolo[l,5- o]pyrimidin-7-yl(isopropyl)carbamate (50 mg, 51%) as a yellow solid. Ή NMR (CDC13, 400 MHz) δ: 10.30 (s, 1H), 8.51 (s, 1H), 7.63 (d, 1H, 7=3.6 Hz), 7.08 (s, 1H),6.88 (dd, 1H,J=3.6, 1.0 Hz), 4.68 (sep, 1H, J= 6.8 Hz), 2.60 (s, 3H), 1.30 (s,9H), 1.28(s,3H), 1.26 (s,3H). LCMS (ES): >95% pure, m/z 301 [M+H-Boc]+.
Example 79. Synthesis of fe -butyl 3-formyl-5-("5-methylthiophen-2-vnpyrazolo[l,5- alpyrimidin-7-yinsopropyncarbamate
Figure imgf000281_0001
Tert-buty\ 3-formyl-5-(5-rnethylthiophen-2-yl)pyrazolo[l ,5-a]pyrirnidin-7- yl(isopropyl)carbamate (48 mg, 0.12 mmol) was dissolved in dichloromethane (0.6 mL).
Trifluoroacetic acid (0.6 mL) was added and the solution changed color from bright yellow to dark orange/red. After 1 h, the solution was concentrated under a stream of air to an orange oil. Trituration with Et20 (3 mL) gave 7-(isopropylamino)-5-(5-methylthiophen-2-yl)pyrazolo[l ,5- a]pyrimidine-3-carbaldehyde (28 mg, 77%) as a tan solid. Ή NMR (DMSO-D6, 400 MHz) δ: 10.01 (s, 1 H), 8.50 (s, 1H), 8.12 (d, 1 H, J= 8.8 Hz), 7.94 (d, 1H, J= 3.6 Hz), 6.95 (s, 1H), 6.92 (dd, 1 H, J= 3.6, 1.0 Hz), 4.14-4.28 (m, 1H), 2.52 (s, 3H), 1.33 (s, 3H), 1.31 (s, 3H). LCMS (ES): >95% pure, m/z 301 [M+H]+.
Example 80. Synthesis of (Z)-5-((7-(isopropylamino)-5-(5-methylthiophen-2-yl)pyrazolo[L5- alpyrimidin-3-yl)methylene)imidazolidine-2,4-dione
Figure imgf000281_0002
7-(Isopropylamino)-5-(5-methylthiophen-2-yl)pyrazolo[l ,5-a]pyrimidine-3-carbaldehyde (20 mg, 0.066 mmol) was suspended in EtOH (0.7 mL). Hydantoin (10 mg, 0.1 mmol) and piperidine (17 μί, 0.165 mmol) were added and the reaction was heated to 80 °C. After 22 h, the solution was filtered while warm and the filter cake was washed with warm EtOH (3 mL) to afford compound (Z)-5-((7-(isopropylamino)-5-(5-methylthiophen-2-yl)pyrazolo[l ,5- a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (22 mg, 88%) as a bright yellow solid. Ή NMR (DMSO-D6, 400 MHz) δ: 1 1.04 (bs, I H), 10.99 (bs, IH), 8.44 (s, IH), 8.05 (d, 1 H, J= 8.8 Hz), 7.90 (d, 1 H, J= 3.6 Hz), 6.95 (d, I H, 7= 3.6), 6.82 (s, 1H), 6.55 (s, I H), 4.14-4.27 (m, IH), 2.53 (s, 3H), 1.33 (s, 3H), 1.32 (s, 3H). LCMS (ES): >95% pure, m/z 383 [M+H]+.
Figure imgf000282_0001
To ethyl 3-(5-methylthiophen-2-yl)-3-oxopropanoate (4.5 g, 21.2 mmol) in AcOH (1 1 mL) was added 3-aminopyrazole (3.25 mL, 21.2 mmol). The reaction mixture was stirred at reflux temperature for 2 hours then cooled to 0°C. The resulting solid was filtered off, washed with EtOAc, and dried under vacuum to provide 3.30 g (67%) of 5-(5-methylthiophen-2- yl)pyrazolo[l ,5-a]pyrimidin-7-ol. (LCMS (ES): >95% pure, m/z 232 [M+H]+. Ή NMR (400 MHz, DMSO-cfe): δ 2.54 (s, 3H), 5 89 (s, I H), 6.19 (d, J=2 Hz, I H), 7.01 (d, J=4 Hz, IH), 7.70 (d, J=4 Hz, IH), 7.88 (s, I H), 12.44 (br s, I H).
Example 82. Synthesis of 7-chloro-5-(5-methylthiophen-2-yl)pyrazolo l ,5-alpyrimidine
Figure imgf000282_0002
To 5-(5-methylthiophen-2-yl)pyrazolo[l ,5-a]pyrimidin-7-ol (3.2 g, 13.8 mmol) in ACN (70 mL) was added phosphorous oxychloride (7.73 mL, 83.0 mmol) and the reaction mixture was heated to reflux temperature. After 6 hours, the solvent was removed by rotary evaporation followed by coevaporation with toluene. The residue was taken up in EtOAc and washed 2x with saturated NaHCC>3. The organic layer was dried with MgSC"4, filtered and the solvent removed to provide 2.88g (83%) of 7-chloro-5-(5-methylthiophen-2-yl)pyrazolo[l ,5- a]pyrimidine. (LC S (ES): >95% pure, m/z 250 [M+H]+. Ή NMR (400 MHz, DMSO-cfe): δ 2.51 (s, 3H), 6.79 (d, J=2 Hz, 1 H), 6.94-6.95 (m, 1H), 7.92 (d, J=6 Hz, 1 H), 7.97 (s, 1 H), 8.27 (d, J=2 Hz, 1 H). Example 83. Synthesis of 7-(benzylthioV5-(5-rnethylthiopheri-2-vnpyrazolori .5-alpyrimidine
Figure imgf000283_0001
To 7-chloro-5-(5-methylthiophen-2-yl)pyrazolo[l ,5-a]pyrimidine (2.88 g, 1 1.5 mmol) in ACN (40 mL) was added triethylamine (2.07 mL, 14.9 mmol) and phenylmethanethiol (1.93 mL, 14.9 mmol). The reaction mixture was stirred at room temperature. After 3 hours, the solvent was removed by rotary evaporation. The residue was taken up in CH2CI2 and washed 2x with water. The organic layer was dried with MgSC , filtered, and the solvent removed to provide 3.25g (84%) of 7-(benzylthio)-5-(5-methylthiophen-2-yl)pyrazolo[l ,5-a]pyrimidine. (LCMS (ES): >95% pure, m/z 338 [M+H]+. Ή NMR (400 MHz, DMSO- 6): 6 2.51 (s, 3H), 4.68 (s, 2H), 6.64 (d, J=2 Hz, 1 H), 6.95-6.96 (m, 1 H), 7.32 (d, J=7Hz, 1 H), 7.37-7.41 (m, 2H), 7.48 (s, 1 H), 7.58 (d, J=7Hz, 2H), 7.89 (d, J=4Hz, 1 H), 8.17 (d, J=2Hz, 1 H).
Example 84. Synthesis of 7-(benzylthioV5-(5-methylthiophen-2-yl)pyrazolo[l ,5-alpyrimidirie- 3-carbaldehyde
Figure imgf000283_0002
To 7-(benzylthio)-5-(5-methylthiophen-2-yl)pyrazolo[l ,5-a]pyrimidine (610 mg, 1.81 mmol) in DMF (4 mL) cooled to 0°C was added phosphorous oxychloride (337 DL, 3.62 mmol). The reaction mixture was stirred while warming to room temperature. After 5 hours, the mixture was cooled to 0°C and 2M NaOH (20 mL) was added slowly while stirring. After stirring for 1 h while warming to rt, the resulting solid was filtered off, washed with water, and dried under vacuum to provide 462 mg (70%) of 7-(benzylthio)-5-(5-methylthiophen-2-yl)pyrazolo[l ,5- a]pyrimidine-3-carbaldehyde. (LCMS (ES): >95% pure, m/z 366 [M+H]+. Ή NMR (400 MHz, DMSO-<¾: δ 2.55 (s, 3H), 4.74 (s, 2H), 7.02-7.03 (m, 1 H), 7.32-7.34 (m, 1 H), 7.38-7.42 (m, 2H), 7.37-7.41 (m, 2H), 7.59 (d, J=8Hz, 2H), 7.73 (s, 2H), 8.06 (d, J=4Hz, 1 H), 8.63 (s, 1 H), · 10.07 (br s, 1 H). Example 85. Synthesis of (Z")-5-((7-(benzylthio")-5-(5-methylthiophen-2-yl)pyrazolori .5- alpyrimidin-3-yDmethylene imidazolidine-2.4-dione
Figure imgf000284_0001
To 7-(benzylthio)-5-(5-methylthiophen-2-yl)pyrazolo[ 1 ,5-a]pyrimidine-3-carbaldehyde
(462 mg, 1.26 mmol) in EtOH (15 mL) was added hydantoin (189 mg, 1.89 mmol), and piperidine (250 uL, 2.52 mmol). The reaction mixture was stirred at 90°C for 5h. The flask was cooled to 0°C and the resulting solid was filtered off and washed with water followed by 1 : 1 EtOH/water. The solid was dried under vacuum to provide 462 mg (70%) of 7-(benzylthio)-5- (5-methylthiophen-2-yl)pyrazolo[l ,5-a]pyrimidine-3-carbaldehyde. (LCMS (ES): >95% pure, m/z 450 [M+H]+. Ή NMR (400 MHz, DMSO-c/6): δ 2.55 (s, 3H), 4.73 (s, 2H), 6.59 (s, 1 H), 7.03 (d, J=4 Hz, 1H), 7.30-7.34 (m, 1H), 7.38-7.42 (m, 2H), 7.58-7.60 (m, 3H), 8.00 (d, J=4Hz, 1 H), 8.63 (s, 1 H), 10.54 (br s, 1 H), 1 1.14 (br s, 1 H).
Figure imgf000284_0002
a]pyrimidin-3-yDmethylene irnidazolidine-2,4-dione
Figure imgf000285_0001
To (Z)-5-((7-(benzylthio)-5-(5-methylthiophen-2-yl)pyrazolo[l ,5-a]pyrirnidin-3- yl)methylene)imidazolidine-2,4-dione (440 mg, 0.98 mmol) in CH2CI2 (20 mL) was added meta- chloroperoxybenzoic acid (848 mg, 4.91 mmol). The reaction mixture was stirred at rt for 16 h. The solid was filtered off and washed with CH2CI2. The material was dried under vacuum to provide 225 mg (49%) of (Z)-5-((7-(benzylsulfinyl)-5-(5-methylthiophen-2-yl)pyrazolo[l ,5- a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione. (LCMS (ES): >95% pure, m/z 464
[M+H]+. Ή NMR (400 MHz, DMSO-i/6): δ 2.53 (s, 3H), 4.67 (dd, J=100,13 Hz, 2H), 6.66 (s, I H), 6.93-6.96 (m, 3H), 7.24-7.25 (m, 4H), 7.81 (d, J=3Hz, I H), 8.84 (s, IH), 10.51 (br s, I H), 1 1.24 (br s, I H).
Example 87. Synthesis of (ZV5-((7-(cvclobutylamino)-5-(5-methylthiophen-2-yl")pyrazolo l .5- alpyrimidin-3-yl')methylene)imidazolidine-2.4-dione
Figure imgf000285_0002
To (Z)-5-((7-(benzylsulfinyl)-5-(5-methylthiophen-2-yl)pyrazolo[l ,5-a]pyrimidin-3- yl)methylene)imidazolidine-2,4-dione (20 mg, 0.04 mmol) in NMP (1.2 mL) was added cyclobutanamine (12 mg, 0.17 mmol). The reaction mixture was stirred at 50°C. After 16 hours, the crude material was filtered through a PTFE filter and purified by mass-directed LC MS to provide (Z)-5-((7-(cyclobutylamino)-5-(5-methylthiophen-2-yl)pyrazolo[l ,5-a]pyrimidin-3- yl)methylene)imidazolidine-2,4-dione as the TFA salt. (LCMS (ES): >95% pure, m/z 395
[M+H]+. Ή NMR (400 MHz, DMSO-<4): δ 1.70-1.80 (m, 2H), 2.30-2.40 (m, 2H), 2.40-2.50 (m, 2H), 2.52 (s, 3H), 4.38-4.50 (m, 1 H), 6.55 (s, 1 H), 6.71 (s, 1H), 6.96 (d, J=4Hz, 1H), 7.89 (d, J=4Hz, 1 H), 8.45 (s, 1 H), 8.57 (d, J=8Hz, 1 H), 10.96 (br s, 1H), 1 1.04 (br s, 1 H).
Example 88. Synthesis of (ZV5-((7-(cvclohexylamino')-5-(5-methylthiophen-2-vnpyrazolo)'1.5- a1pyrimidin-3-yQmethylene)imidazolidine-2,4-dione
Figure imgf000286_0001
Same procedure as Example 87. LCMS (ES):>90% pure, m/z 423 [M+H]+. Ή NMR (400 MHz, DMSO-c/e): δ 1.10-1.20 (m, 1 H), 1.40-1.60 (m, 4H), 1.60-1.70 (m, 1 H), 1.70-1.80 (m, 2H), 1.90-2.00 (m, 2H), 2.52 (s, 3H), 3.80-3.90 (m, 1H), 6.55 (s, 1 H), 6.83 (s, 1H), 6.96 (dd, J=4,l Hz, 1 H), 7.93 (d, J=4Hz, 1 H), 8.00 (d, J=8Hz, 1 H), 8.43 (s, 1 H), 10.98 (br s, 1 H), 1 1.03 (br s, 1 H). Example 89. Synthesis of (Z)-5-((5-(5-methylthiophen-2-vQ-7-(2-morpholino
propylamino')pyrazolori .5-alpyrimidin-3-ynmethylene imidazolidine-2,4-dione
Same procedure as Example 87. LCMS (ES):>90% pure, m/z 468 [M+H]+. Ή NMR (400 MHz, DMSO-i e): δ 1.37 (d, J=6Hz, 3H), 2.54 (s, 3H), 3.20-3.40 (m, 2H), 3.40-3.60 (m, 2H), 3.70-3.90 (m, 4H), 3.90-4.00 (m, I H), 4.00-4.20 (m, 2H), 6.57 (s, I H), 6.85 (s, I H), 6.99 (dd, J=4,l Hz, I H), 7.91 (d, J=4Hz, IH), 8.51 (s, IH), 8.58 (br s, I H), 9:80 (br s, I H), 10.90 (d, J=l Hz, IH), 1 1.08 (br s, IH).
Example 90. Synthesis of (Zy5-(f7-f2-ethoxyethylaminoV5-(5-methylthiophen-2- vDpyrazolofl .S-alpyrimidin^-vnmethylene midazolidine^^-dione
Figure imgf000287_0002
Same procedure as Example 87. LCMS (ES):>90% pure, m/z 413 [M+H]+. Ή NMR (400 MHz, DMSO-i/e): δ 1.09 (t, J=7Hz, 3H), 2.55 (s, 3H), 3.49 (q, J=7Hz, 2H), 3.60-3.80 (m, 4H), 6.55 (s, I H), 6.86 (s, IH), 6.96 (d, J=4Hz, IH), 7.87 (d, J=4Hz, I H), 8.24 (t, J=6Hz, I H), 8.44 (s, I H), 10.99 (s, IH), 1 1.04 (s, IH).
Example 91 . Synthesis of (ZVS-^-^-fdimethylaminolethylaminoVS-CS-methylthiophen - 2- yl pyrazolon ,5-a1pyrimidin-3-yl)methylene)imidazolidine-2,4-dione
Figure imgf000288_0001
Same procedure as Example 87. LCMS (ES):>90% pure, m/z 412 [M+H]+. Ή NMR (400 MHz, DMSO- ): δ 2.55 (s, 3H), 2.89 (s, 6H), 3.40-3.50 (m, 2H), 3.80-3.90 (m, 2H), 6.57 (s, IH), 6.88 (s, IH), 6.99 (dd, J=4, IHz, IH), 7.91 (d, J=4Hz, IH), 8.49 (t, J=4Hz, IH), 9.42 (br s, IH), 10.91 (s, IH), 11.07 (s, IH).
Example 92. Synthesis of (Zy5-((5-(5-methylthiophen-2-y0-7-(2-morpholino
ethylamino")pyrazolo[1.5-alpyrimidin-3-yl")methylene imidazolidine-2,4-dione
Figure imgf000288_0002
Same procedure as Example 87. LCMS (ES):>90% pure, m/z 454 [M+H]+. Ή NMR (400 MHz, DMSO-c4): δ 2.53 (s, 3H), 3.10-3.30 (m, 2H), 3.49 (br s, 2H), 3.60-3.80 (m, 4H), 3.90-4.00 (m, 2H), 4.00-4.10 (m, 2H), 6.57 (s, IH), 6.89 (s, IH), 6.99 (dd, J=4, IHz, IH), 7.91 (d,J=4Hz, IH), 8.53 (t, J=8Hz, IH), 9.78 (br s, IH), 10.91 (s, IH), 11.07 (s, IH).
Example 93. Synthesis of (ZV5-((7-(cvclopropylmethylaminoV5-(5-methylthiophen-2- vnpyrazolori.S-alpyrimidin-S-ynmethylene^midazolidine^^-dione
Figure imgf000289_0001
Same procedure as Example 87. LCMS (ES):>90% pure, m/z 395 [M+H]+. Ή NMR (400 MHz, DMSO-cfe): δ 0.23-0.31 (m, 2H), 0.40-0.50 (m, 2H), 2.53 (s, 3H), 3.40-3.50 (m, 2H), 6.56 (s, I H), 6.84 (s, I H), 6.96 (dd, J=4, I Hz, I H), 7.91 (d, J=4Hz, I H), 8.42 (t, J=6Hz, I H), 1 1.00 (s, IH), 1 1.03 (s, IH).
Example 94. Synthesis of ("ZV5-('('7-(benzylamino -5-('5-methylthiophen-2-ynpyrazolo[l ,5- a1pyrimidin-3-yl")methylene)imidazolidine-2,4-dione
Figure imgf000289_0002
Same procedure as Example 87. LCMS (ES):>90% pure, m/z 431 [M+H]+. Ή NMR (400 MHz, DMSO-i/e): δ 2.53 (s, 3H), 4.74 (d, J=6Hz, 2H), 6.55 (s, 1 H), 6.75 (s, 1 H), 6.93 (dd, J=4, I Hz, I H), 7.20-7.30 (m, I H), 7.30-7.40 (m, 2H), 7.40-7.50 (m, 2H), 7.76 (d, J=3Hz, I H), 9.00 (t, J=6Hz, I H), 10.95 (s, I H), 1 1.03 (s, I H).
Example 95. Synthesis of (ZV5-(Y7-(2-hydroxyethylaminoy5-(5-methylthiophen-2- ynpyrazolori ,5-alpyrimidin-3-vnmethylene)imidazolidine-2,4-dione
Figure imgf000290_0001
Same procedure as Example 87. LCMS (ES):>90% pure, m/z 385 [M+H]+. Ή NMR (400 MHz, DMSO-c/6): δ 2.52 (s, 3H), 3.50-3.60 (m, 2H), 3.60-3.70 (m, 2H), 6.55 (s, I H), 6.83 (s, I H), 6.96 (dd, J=4, I Hz, I H), 7.85 (d, J=4Hz, IH), 8.17 (t, J=6Hz, 2H), 8.43 (s, I H), 1 1.00 (s, I H), 1 1.03 (s, I H).
Example 96. Synthesis of (Z)-5-((7-(2-(2-.6-dimethylmorpholino')ethylamino')-5-(5- methylthiophen-2-yl)pyrazolo[l ,5-a1pyrimidin-3-ynmethylene)imidazolidine-2,4-dione
Figure imgf000290_0002
Same procedure as Example 87. LCMS (ES):>90% pure, m/z 482 [M+H]+. Ή NMR (400 MHz, DMSO-efe): δ 1.15 (d, J=6Hz, 6H), 2.53 (s, 3H), 2.60-2.80 (m, 2H), 3.20-3.40 (m, 2H), 3.54 (br s, 2H), 3.82 (br s, 2H), 3.91 (br s, 2H), 6.57 (s, I H), 6.89 (s, I H), 6.99 (d, J=4Hz, 1 H), 7.92 (d, J=4Hz, 1 H), 8.50 (s, 1 H), 9.80 (br s, 1 H), 10.90 (s, 1 H), 1 1 .07 (s, 1 H). Example 97. Synthesis of ("ZV5-(("5-(5-methylthiophen-2-yl")-7-(3-(pyrrolidin-l - vnpropylamino'lpyrazolof l .S-alpyrimidin-S-vnmethylene midazolidine^^-dione
Figure imgf000291_0001
Same procedure as Example 87. LCMS (ES):>90% pure, m/z 452 [M+H]+. Ή NMR (400 MHz, DMSO-c¼): δ 1 .83 (br s, 2H), 2.00 (br s, 4H), 2.54 (s, 3H), 3.00 (br s, 2H), 3.22 (br s, 2H), 3.50-3.70 (m, 4H), 6.56 (s, I H), 6.81 (s, I H), 6.98 (dd, J=4, I Hz, I H), 7.89 (d, J=4Hz, I H), 8.46 (s, 1 H), 8.49 (s, 1 H), 9.50 (br s, 1 H), 10.96 (s, 1 H), 1 1.05 (s, 1 H).
Example 98. Synthesis of (ZV5-('('7-(4-fluorophenylaminoV5-('5-methylthiophen-2- vnpyrazolori ,5-alpyrimidin-3-vnmethylene)imidazolidine-2.4-dione
Figure imgf000291_0002
To (Z)-5-((7-(benzylsulfinyl)-5-(5-methylthiophen-2-yl)pyrazolo[l ,5-a]pyrimidin-3- yl)methylene)imidazolidine-2,4-dione (20 mg, 0.04 mmol) in isopropanol (2 mL) was added p- toluenesulfonic acid (8 mg, 0.04 mmol) and 4-fluoroaniline (22 mg, 0.2 mmol). The reaction mixture was stirred under microwave heating at 1 10°C for 2 hours, filtered through a PTFE filter , and purified by mass-directed LC/MS to provide (Z)-5-(f7-f4-fluorophenylamino)-5-(5- methylthiophen-2-yl)pyrazolo[l ,5-a] as the
TFA salt. (LCMS (ES): >95% pure, m/z 435 [M+H]+. Ή NMR (400 MHz, DMSO-</6): δ 2.53 (s, 3H), 6.59 (s, 1 H), 6.60 (s, 1 H), 6.89 (dd, J=4, lHz, 1H), 7.30-7.40 (m, 2H), 7.50-7.70 (m, 3H), 8.58 (s, 1H), 10.32 (s, 1 H), 10.92 (s, 1 H), 1 1.08 (s, 1 H).
Example 99. Synthesis of ethyl-6-(4-nitrophenoxy)hexanoate
Figure imgf000292_0001
To 4-nitrophenol (2 g, 14.4 mmol) in DMF (25 mL) was added ethyl-6-bromohexyanoate
(3.21 g, 14.4 mmol) and 2C03 (3.97 g, 28.8 mmol). The reaction mixture was stirred at 50°C. After 3h, the flask was cooled to 0°C and the solid was filtered off and washed with water. The material was dried under vacuum to provide 3.6g (89%) of ethyl-6-(4-nitrophenoxy)hexanoate. Example 100. Synthesis of ethyl 6-(4-aminophenoxy')hexanoate
Figure imgf000292_0002
To ethyl 6-(4-nitrophenoxy)hexanoate (3.63 g, 12.9 mmol) in EtOH (50 mL) was added 10% palladium on carbon (600 mg). The reaction mixture was stirred at rt for 16h under 1 atm of hydrogen. The material was filtered through a pad of celite and the filtrate was concentrated under rotary evaporation to provide 3.2g (100%) of ethyl 6-(4-aminophenoxy)hexanoate.
Example 101. Synthesis of (Z)-ethyl 6-(4-(3-((2.5-dioxoimidazolidin-4-ylidene)methyl")-5-C5- methylthiophen-2-vnpyrazolo 1.5-a1pyrimidin-7-ylamino)phenoxy')hexanoate
Figure imgf000293_0001
To (Z)-5-((7-(benzylsulfmyl)-5-(5-methylthiophen-2-yl)pyrazolo[l ,5-a]pyrimidin-3- yl)methylene)imidazolidine-2,4-dione (20 mg, 0.04 mmol) in isopropanol (1 mL) was added p- toluenesulfonic acid (8 mg, 0.04 mmol) and ethyl 6-(4-aminophenoxy)hexanoate (54 mg, 0.21 mmol). The reaction mixture was stirred under microwave heating at 1 10°C for 20 min. The reaction mixture was cooled to 0°C and the resulting solid was filtered off and washed with water followed by isopropanol. The material was dried under vacuum to provide (Z)-ethyl 6-(4-(3- ((2,5-dioxoimidazolidin-4-ylidene)methyl)-5-(5-methylthiophen-2-yl)pyrazolo[l ,5-a]pyrimidin- 7-ylamino)phenoxy)hexanoate. (LCMS (ES): >95% pure, m/z 575 [M+H]+.
Example 102. Synthesis of methyl 2-(4-nitrophenoxy)acetate
Figure imgf000293_0002
Same procedure as Example 99. LCMS (ES):>90% pure, m/z 212 [M+H]+.
Example 103. Synthesis of /e -butyl cvclopropyl(3-formyl-5-(thiophen-2-yl)pyrazolo 1.5- <7]pyrimidin-7-yl)carbamate
Figure imgf000294_0001
DME and 2M a2CC>3 were degassed with a stream of nitrogen for 10 minutes prior to use. 7e -butyl 5-chloro-3-formylpyrazolo[l ,5-cr]pyrimidin-7-yl(cyclopropyl)carbamate (672 mg, 2 mmol) and thiophene-2-boronic acid (384 mg, 3 mmol) were dissolved in DME (12 mL) under an atmosphere of nitrogen. 2M Na2C03 (3 mL), and Pd(PPli3)4 (231 mg, 0.2 mmol) were added sequentially, and the reaction was placed in a 90°C oil bath. After 18 h, the solution was partitioned between EtOAc (75 mL) and 0.5N HC1 (75 mL). The aqueous layer was again extracted with EtOAc (75 mL) and the organics were washed with brine (150 mL), dried over MgSC>4, filtered, and concentrated in vacuo. The brown residue was purified via flash column chromatography (10-20% EtOAc/hexanes) to provide teri-butyl cyclopropyl(3-formyl-5- (thiophen-2-yl)pyrazolo[l ,5-a]pyrimidin-7-yl)carbamate (646 mg, 84%) as a yellow solid. Ή NMR (CDC13, 400 MHz) δ: 10.34 (s, 1H), 8.53 (s, 1H), 7.82 (dd, 1 H, J= 4.0, 1.2 Hz), 7.63 (dd, 1H, J= 5.2, 1.2 Hz), 7.21 (dd, 1H, J= 5.2, 4.0 Hz), 7.18 (s, 1 H), 3.30 (dddd, 1 H, 7= 7.2, 7.2, 3.2, 3.2 Hz), 1.41 (s, 9H), 0.81 -0.95 (m, 2H), 0.61-0.74 (m, 2H). LCMS (ES): >95% pure, m/z 485 [M+H]+.
Example 104. Synthesis of 7-(cvclopropylamino)-5-(thiophen-2-ynpyrazolo[K5-q]pyrimidine- 3-carbaldehyde
Figure imgf000294_0002
7¾r/-butyl cyclopropyl(3-formyl-5-(thiophen-2-yl)pyrazolo[l ,5-a]pyrimidin-7- yl)carbamate (457 mg, 1.19 mmol) was dissolved in dichloromethane (6 mL). Trifluoroacetic acid (6 mL) was added and after 1 h, the solution was concentrated under a stream of air to a dark red/brown oil. Trituration with Et20 (20 mL) gave 7-(cyclopropylamino)-5-(thiophen-2- yl)pyrazolo[l ,5-a]pyrimidine-3-carbaldehyde (248 mg, 73%) as a light orange solid. 'H NMR (DMSO-D6, 400 MHz) δ: 10.04 (s, 1 H), 8.83 (d, 1H, J= 2.0 Hz), 8.53 (s, 1H), 8.06 (dd, 1H, J = 4.0, 0.8 Hz), 7.82 (dd, 1 H, J= 5.2, 0.8 Hz), 7.25 (dd, 1 H, J= 5.2, 4.0 Hz), 7.05 (s, 1 H), 2.80-2.88 (m, 1 H), 0.87-1.00 (m, 2H), 0.75-0.86 (m, 2H). LCMS (ES): >95% pure, m/z 285 [M+H]+.
Example 105. Synthesis of (Z)-5-((7-(cvclopropylaminoV5-(thiophen-2-vnpyrazolor i .5- q pyrimidin-3-yl")methylene)imidazolidine-2,4-dione
Figure imgf000295_0001
(Z)-5-((7-(Cyclopropylamino)-5-(thiophen-2-yl)pyrazolo[l ,5-cr]pyrimidin-3- yl)methylene)thiazolidine-2,4-dione (35 mg, 0.12 mmol) was suspended in EtOH (1.2 mL). Hydantoin (19 mg, 0.18 mmol) and piperidine (19 μΐ^, 0.18 mmol) were added and the reaction was heated to 80 °C. After 16 h, the solution was filtered while warm and the filter cake was washed with warm EtOH (4 mL) to afford compound (Z)-5-((7-(cyclopropylamino)-5-(thiophen- 2-yl)pyrazolo[l ,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (40 mg, 88%) as a bright yellow solid. Ή NMR (DMSO-D6, 400 MHz) δ: 1 1.05 (bs, 1 H), 1 1.02 (bs, 1 H), 8.76 (bs, 1 H), 8.44 (s, 1 H), 8.03 (dd, 1 H, J= 3.8, 1.0 Hz), 7.87 (dd, 1 H, 7= 4.8, 1.0 Hz), 7.27 (dd, 1 H, 7 = 4.8, 3.8 Hz), 6.93 (s, 1 H), 6.56 (s, 1 H), 2.81 -2.89 (m, 1 H), 0.87-1.00 (m, 2H), 0.73-0.85 (m, 2H). LCMS (ES): >95% pure, m/z 367 [M+H]+.
Table 13. Biological Activities of Examples 68, 74, 80, 85 to 98, 101 , and 105.
Figure imgf000295_0002
Figure imgf000296_0001
Example 106. Synthesis of 5,7-dichloro-6-methylpyrazolo[l .5-alpyrimidine
Figure imgf000296_0002
Under nitrogen gas atmosphere, sodium (3.5 g, 150 mmol) was added to ethanol (125 mL) in small portions and stirred at room temperature until all the sodium had dissolved. A solution of 3-aminopyrazole (13 g, 150 mmol) in ethanol (20 mL) and diethyl methylmalonate (26 mL, 150 mmol) were dropped, successively, to the above solution. The mixture was refluxed at 90°C for 10 hours, cooled to room temperature, and filtered under vacuum. To the solid, cold 5N HC1 was added and the resulting solid was collected by filtration under vacuum. The intermediate, 6-methylpyrazolo[l ,5-a]pyrimidine-5,7-diol, was recovered as an off-white solid in 72% yield (18 g). This material was used for the next step without further purification. LCMS (M+l=166)
Under nitrogen gas atmosphere, phosphorous oxychloride (160 mL, 1 .7 mol) and dimethylanihne (16 mL, 130 mmol) was added successively to the intermediate prepared above (16 g, 100 mmol). The mixture was heated at 1 10°C for 4 hours then excess POC was removed under vacuum. The residue was made basic with 3N NaOH solution (pH = 9-10) and extracted with ethyl acetate (3x). The combined organic layers were dried over anhydrous Na2SC>4, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (100% DCM) to provide 16 grams of the solid yellow product, 5,7-dichloro-6-methylpyrazolo[l ,5- a]pyrimidine (81 % yield). LCMS (M+l =203)
Example 107. Synthesis of 5-chloro-7-(cvclopropylamino)-6-methylpyrazolo l ,5-alpyrimidine- 3-carbaldehyde
Figure imgf000297_0001
To the reaction flask, 5,7-dichloro-6-methylpyrazolo[l ,5-a]pyrimidine (5 g, 25 mmol) was added along with cyclopropyl amine ( 1 .8 mL, 25 mmol), triethylamine (3.5 mL, 25 mmol), and acetonitrile (90 mL). The reaction was stirred at room temperature for 3 hours then heated at 85°C for an additional 6 hours. The mixture was cooled to room temperature, diluted with water, filtered and washed with water. The intermediate, 5-chloro-N-cyclopropyl-6- methylpyrazolo[l ,5-a]pyrimidin-7-amine, was further purified by silica gel chromatography (10% ethyl acetate/hexanes) to provide 4.8 grams of a white solid (86% yield). LCMS (M+l = 223) To the intermediate (3.6 g, 16 mmol) isolated above in DMF (60 mL) was added phosphorous oxychloride (9 mL, 100 mmol) slowly at room temperature. The reaction mixture was allowed to stir at room temperature for 10 hours then quenched by addition to 6N NaOH solution. The pH of the mixture was adjusted with 6N HC1 to pH = 7-9. The solid was recovered by filtration and washed with water. The product, 5-chloro-7-(cyclopropylamino)-6- methylpyrazolo[l,5-a]pyrimidine-3-carbaldehyde, was purified by recrystallization from ethyl acetate/hexanes to yield a white solid in 73% yield (2.9 g). LCMS (M+l= 251 )
Example 108. Synthesis of tert-butyl 5-chloro-3-formyl-6-methylpyrazolon ,5-alpyrimidin-7- vKcyclopropyPcarbamate
Figure imgf000298_0001
To 5-chloro-7-(cyclopropylamino)-6-methylpyrazolo[l ,5-a]pyrimidine-3-carbaldehyde (2.9 g, 12 mmol) in methylene chloride (22 mL) was added triethylamine (2 mL, 14 mmol), dimethylaminopyridine (100 mg, 0.8 mmol), and di-i-butyldicarbonate (3.1 g, 14 mmol). The mixture was stirred at room temperature for 10 hours. The reaction mixture was transferred to a separatory funnel, washed IX with H2O, 2X with brine, dried over MgSC^, filtered, and evaporated to dryness to provide an oily residue. The crude material was purified by silica gel chromatography (25% ethyl acetate/hexanes) to yield a light orange solid (3.6 g, 88% yield), tert- butyl 5-chloro-3-formyl-6-methylpyrazolo[l ,5-a]pyrimidin-7-yl(cyclopropyl)carbamate. LCMS (M+l = 351)
Example 109. Synthesis of 5-f7-(cvclopropylamino)-3-formyl-6-methylpyrazolon ,5- alpyrimidin-5-yDthiophene-2-carbonitrile
Figure imgf000299_0001
Tert-buty\ 5-chloro-3-formyl-6-methylpyrazolo[l,5-a]pyrimidin-7- yl(cyclopropyl)carbamate (420 mg, 1.2 mmol) was mixed with 5-cyanothiophen-2-ylboronic acid (280 mg, 2 mmol), triphenylphosphine (69 mg, 0.06 mmol), and NaaCC (2N in water, 1.8 mL, 3.6 mmol) in (2: 1 ) DME/EtOH (12 mL). The reaction was heated at 85°C overnight then cooled to room temperature. The reaction was evaporated to dryness, dissolved in DCM, filtered and evaporated to dryness. The residue was then dissolved in (1 : 1) TFA/DCM (4 mL) and stirred at room temperature for 1 hour. Excess solvent and TFA were removed by evaporation under a stream of nitrogen. The residue was dissolved in DCM, washed with saturated NaHCC^ solution (3x) and saturated NaCl solution. The organic layer was isolated, dried over anhydrous MgS04, filtered and concentrated by rotary evaporation The residue was purified by silica gel chromatography (25% EtOAc/hexanes) to provide 5-(7-(cyclopropylamino)-3-formyl-6- methylpyrazolo[l ,5-a]pyrimidin-5-yl)thiophene-2-carbonitrile (47 mg, 12% yield). LCMS (M+ 1=324)
Example 1 10. Synthesis of 5-(7-('cvclopropylamino)-3-((2,5-dioxoimidazolidin-4- ylidene)methyl)-6-methylpyrazolo["l ,5-a1pyrimidin-5-yl)thiophene-2-carbonitrile
Figure imgf000299_0002
Same procedure as Example 4. LCMS m/z 406 [M+H] Example 1 1 1. Synthesis of (Z)-5-((7-(cyclopropylamino)-5-(thiophen-2-vQpyrazolon ,5- a]pyrimidin-3-yl)methylene)thiazolidine-2.4-dione
Figure imgf000300_0001
7-(Cyclopropylamino)-5-(thiophen-2-yl)pyrazolo[l ,5-a]pyrimidine-3-carbaldehyde (35 mg, 0.12 mmol) was suspended in EtOH (1.2 mL). 2,4-thazolidinedione (21 mg, 0.18 mmol) and piperidine (19 μί, 0.18 mmol) were added and the reaction was heated to 80 °C. After 16 h, the solution was filtered while warm and the filter cake was washed with warm EtOH (4 mL) to afford compound (Z)-5-((7-(cyclopropylamino)-5-(thiophen-2-yl)pyrazolo[l ,5-a]pyrimidin-3- yl)methylene)thiazolidine-2,4-dione (32 mg, 68%) as a bright yellow solid. Ή NMR (DMSO- D6, 400 MHz) δ: 12.25 (bs, 1H), 8.75 (d, 1 H, J= 2.4 Hz), 8.33 (s, 1H), 8.04 (d, 1 H, J= 3.2 Hz), 7.89 (s, 1 H), 7.87 (d, 1H, J= 5.2 Hz), 7.25 (dd, 1H, J= 5.2, 3.2 Hz), 6.99 (s, 1 H), 2.80-2.88 (m, 1H), 0.90-1.00 (m, 2H), 0.75-0.84 (m, 2H). LCMS (ES): >95% pure, m/z 384 [M+H]+.
Example 1 12. Synthesis of fe -butyl 5-(5-cyanothiophen-2-vD-3-formylpyrazolo|"l .5- alpyrimidin-7-yl(cvclopropyl)carbamate
Figure imgf000300_0002
DME and 2M Na2C03 were degassed with a stream of nitrogen for 10 minutes prior to use. tert-butyl 5-chloro-3-formylpyrazolo[l ,5-a]pyrimidin-7-yl(cyclopropy!)carbamate (445 mg, 1.32 mmol) and 5-cyanothiophene-2-boronic acid (302 mg, 1.98 mmol) were dissolved in DME (12 mL) under an atmosphere of nitrogen. 2M Na2CC>3 (2 mL), and Pd(PPli3)4 (152 mg, 0.13 mmol) were added sequentially. The reaction was placed in a 90 °C oil bath. After 18 h, the solution was partitioned between EtOAc (75 mL) and 0.5N HC1 (75 mL). The aqueous layer was again extracted with EtOAc (75 mL) and the organics were washed with brine (150 mL), dried over MgSC>4, filtered, and concentrated in vacuo. The dark red/brown residue was purified via flash column chromatography (10-30% EtOAc/hexanes) to provide tert-butyl 5-(5- cyanothiophen-2-yl)-3-formylpyrazolo[l,5-o]pyrimidin-7-yl(cyclopropyl)carbamate (224 mg, 41%) as a foamy yellow solid. Ή NMR (CDC13, 400 MHz) δ: 10.34 (s, 1H), 8.59 (s, 1H), 7.76 (d, l H, J= 3.8 Hz), 7.69 (d, 1 H, J = 3.8 Hz), 7.24 (s, 1 H), 3.34 (dddd, 1H, J= 7.2, 7.2, 3.2, 3.2 Hz), 1.44 (s, 9H), 0.87-0.95 (m, 2H), 0.59-0.72 (m, 2H). LCMS (ES): >95% pure, m/z 410 [M+H]+.
Example 1 13. Synthesis of (Z)-fert-butyl 5-(5-cyanothiophen-2-yl)-3-((2,4-dioxothiazolidin-5- ylidene')methvnpyrazolori ,5-alpyrimidin-7-yl(cyclopropyl')carbamate
Figure imgf000301_0001
7¾r/-butyl 5-(5-cyanothiophen-2-yl)-3-formylpyrazolo[l ,5-a]pyrimidin-7- yl(cyclopropyl)carbamate (102 mg, 0.25 mmol) was suspended in EtOH (2.5 mL). 2,4- thazolidinedione (44 mg, 0.38 mmol) and piperidine (37 μί, 0.38 mmol) were added and the reaction was heated to 80 °C. After 18 h, the solution was diluted with ¾0 (50 mL) and extracted with dichioromethane (3 x 50 mL). The organics were washed with brine (1 x 200 mL), dried over MgSC>4, filtered and concentrated in vacuo to a brown oil. The residue was purified via preparative TLC (45% EtOAc/hexanes) to afford compound (Z)-tert-buty\ 5-(5- cyanothiophen-2-yl)-3-((2,4-dioxothiazolidin-5-ylidene)methyl)pyrazolo[l ,5-a]pyrimidin-7- yl(cyclopropyl)carbamate (47 mg, 37%) as a bright orange solid. Ή NMR (CDC13, 400 MHz) δ: 8.30 (bs, I H), 8.20 (s, IH), 7.74 (d, IH, J= 4.0 Hz), 7.68 (d, 1H, J = 4.0 Hz), 7.17 (s, I H), 3.33 (dddd, I H, J= 6.8, 6.8, 3.6, 3.6 Hz), 1.46 (s, 9H), 0.83-0.94 (m, 2H), 0.61 -0.70 (m, 2H). LCMS (ES): >90% pure, m/z 509 [M+H]+.
Example 1 14. Synthesis of CZ)-5-(7-(cvclopropylamino)-3-((2,4-dioxothiazolidin-5- ylidene)methynpyrazolori .5-a1pyrimidin-5-yl")thiophene-2-carbonitrile
Figure imgf000302_0001
(Z)-Tert-buty\ 5-(5-cyanothiophen-2-yl)-3-((2,4-dioxothiazolidin-5- ylidene)methyl)pyrazolo[l ,5-a]pyrimidin-7-yl(cyclopropyl)carbamate (40 mg, 0.08 mmol) was dissolved in dichloromethane (0.8 mL) and trifluoroacetic acid (0.8 mL). After 1 h, the solution was concentrated under a stream of air. Trituration of the residue with Et20 (5 mL) gave (Z)-5- (7-(cyclopropylamino)-3-((2,4-dioxothiazolidin-5-ylidene)methyl)pyrazolo[l ,5-a]pyrimidin-5- yl)thiophene-2-carbonitrile (21 mg, 62%) as a bright yellow/orange solid. Ή NMR (DMSO-D6, 400 MHz) δ: 12.30 (bs, I H), 8.88 (d, lH, J= 2.8 Hz), 8.33 (s, IH), 8.16 (d, lH, J= 4.2 Hz), 8.06 (d, I H, J = 4.2 Hz), 7.83 (s, I H), 7.10 (s, I H), 2.80-2.88 (m, I H), 0.90-0.97 (m, 2H), 0.77-0.84 (m, 2H). LCMS (ES): >95% pure, m/z 409 [M+H]+.
Example 1 15. Synthesis of methyl 5-(7-(fer/-butoxycarbonvi cvclopropynamino)-3- formylpyrazolo 1.5-a]pyrimidin-5-ynthiophene-2-carboxylate
Figure imgf000303_0001
DME and 2M Na2CC>3 were degassed with a stream of nitrogen for 10 minutes prior to use. Tert-butyl 5-chloro-3-formylpyrazolo[l,5-a]pyrimidin-7-yl(cyclopropyl)carbamate (672 mg, 2 mmol) and 5-(methoxycarbonyl)thiophene-2-boronic acid pinacol ester (804 mg, 3 mmol) were dissolved in DME (12 mL) under an atmosphere of nitrogen. 2M Na2C03 (3 mL), and Pd(PPli3)4 (231 mg, 0.2 mmol) were added sequentially. The reaction was placed in a 90 °C oil bath. After 4 h, the solution was partitioned between EtOAc (75 mL) and 0.5N HC1 (75 mL). The aqueous layer was further extracted with EtOAc (2 x 75 mL) and the organics were washed with brine (100 mL), dried over MgS04, filtered, and concentrated in vacuo. The residue was purified via flash column chromatography (10-30% EtOAc/hexanes) to provide methyl 5-(7- (iert-butoxycarbonyl(cyclopropyl)amino)-3-formylpyrazolo[l ,5-a]pyrimidin-5-yl)thiophene-2- carboxylate (188 mg, 21%) as an orange solid. Ή NMR (CDCI3, 400 MHz) δ: 10.35 (s, 1 H), 8.57 (s, 1 H), 7.85 (d, 1H, 7= 4.2 Hz), 7.78 (d, 1 H, J= 4.2 Hz), 7.23 (s, 1 H), 3.96 (s, 3H), 3.32 (dddd, 1 H, J= 6.8, 6.8, 3.6, 3.6 Hz), 1.43 (s, 9H), 0.85-0.94 (m, 2H), 0.63-0.69 (m, 2H). LCMS (ES): >95% pure, m/z 443 [M+H]+.
Example 1 16. Synthesis of methyl 5-(7-(cvclopropylamino)-3-formylpyrazolori ,5-a1pyrimidin- 5-yl)thiophene-2-carboxylate
Figure imgf000303_0002
5-(7-(rert-butoxycarbonyl(cyclopropyl)amino)-3-formylpyrazolo[l ,5-iJf]pyrimidin-5- yl)thiophene-2-carboxylate (173 mg, 0.39 mmol) was dissolved in dichioromethane (3 mL) and trifluoroacetic acid (3 mL). After 1 h, the solution was concentrated under a stream of air.
Trituration of the residue with Et20 (10 mL) and then MeOH (5 mL) gave methyl 5-(7- (cyclopropylamino)-3-formylpyrazolo[l,5-a]pyrimidin-5-yl)thiophene-2-carboxylate (98 mg, 74%) as an orange solid. Ή NMR (DMSO-D6, 400 MHz) δ: 10.06 (s, 1H), 8.99 (d, 1 H, J= 2.4 Hz), 8.57 (s, lH), 8.13 (d, 1 H, J= 4.0 Hz), 7.88 (d, 1 H, J= 4.0 Hz), 7.16 (s, 1 H), 3.88 (s, 3H), 2.81-2.90 (m, 1 H), 0.90-0.99 (m, 2H), 0.76-0.84 (m, 2H). LCMS (ES): >90% pure, m/z 343 [M+H]+.
Example 1 17. Synthesis of (Z)-methyl 5-(7-(cyclopropylamino)-3-((2,4-dioxothiazolidin-5- ylidene)methyl)pyrazolo[K5-a]pyrimidin-5-yl)thiophene-2-carboxylate
Figure imgf000304_0001
Methyl 5-(7-(cyclopropylamino)-3-formylpyrazolo[l ,5-a]pyrimidin-5-yl)thiophene-2- carboxylate (35 mg, 0.10 mmol) was suspended in EtOH (1 mL). 2, 4-thazolidinedione (18 mg, 0.15 mmol) and piperidine (16 μί, 0.15 mmol) were added and the reaction was heated to 80 °C. After 23 h, the solution was filtered while it was still warm and the solid was washed with warm EtOH (5 mL). The solid was triturated with dichloromethane (2 x 3 mL) to furnish compound (Z)-methyl 5-(7-(cyclopropylamino)-3-((2,4-dioxothiazolidin-5-ylidene)methyl)pyrazolo[l ,5- fl]pyrimidin-5-yl)thiophene-2-carboxylate (30 mg, 67%) as an orange solid. Ή NMR (DMSO- D6, 400 MHz) δ: 12.30 (bs, 1 H), 8.92 (d, 1 H, J= 2.4 Hz), 8.35 (s, 1 H), 8.10 (d, l H, J= 4.4 Hz), 7.88 (d, 1 H, J = 4.4 Hz), 7.87 (s, 1H),7.08 (s, 1 H), 3.88 (s, 3H), 2.81-2.90 (m, 1 H), 0.90-0.99 (m, 2H), 0.76-0.84 (m, 2H). LCMS (ES): >95% pure, m/z 442 [M+H]+. Table 14. Biological Activities of Examples 1 10, 1 1 1 , 1 13, 1 14, and 1 17.
Figure imgf000305_0001
Example 1 18. Synthesis of fZ>N-f3-(7-fcyclopropylamino>3-rd-(hvdroxym
dioxoimidazolidin^-ylidene^methvDpyrazolofLS-alpyrimidin-S-ynphenvnmethanesulfonamide
Figure imgf000305_0002
To (Z)-N-(3-(7-(cyclopropylamino)-3-((2,5-dioxoimidazolidin-4- ylidene)methyl)pyrazolo[l ,5-a]pyrimidin-5-yl)phenyl)methanesulfonamide (200 mg, 0.44 mmol) in acetonitrile (4 mL) and pyridine (400 uL) was added formaldehyde (37% aq) (1.5 mL). The reaction mixture was stirred at 65°C for 15 minutes. Cooled to rt and removed half the volume of solvent by rotary evaporation. Added water (5 mL) and sonicated until solid formed. The resulting solid was filtered off and washed with water. Dried under vacuum to provide 185 mg (87%) of (Z)-N-(3-(7-(cyclopropylamino)-3-((l-(hydroxymethyl)-2,5-dioxoimidazolidin-4- ylidene)methyl)pyrazolo[l ,5-a]pyrimidin-5-yl)phenyl)methanesulfonamide. LCMS (ES): >95% pure, m/z 484 [M+l ]+.
Example 1 19. Synthesis of (S.Z')-f4-r('7-(cyclopropylamino')-5-(3-
(methylsulfonamido'lphenvnpyrazolori .S-alpyrimidin^-vnmethylene^^.S-dioxoimidazolidin-l - vQmethyl 2-("tert-butoxycarbonylamino')propanoate
Figure imgf000306_0001
To (Z)-N-(3-(7-(cyclopropylamino)-3-((l -(hydro
ylidene)methyl)pyrazolo[l ,5-a]pyrimidin-5-yl)phenyl)methanesulfonarnide (90 mg, 0.19 mmol) in DMF (2 mL) cooled to 0°C was added Boc-L-Ala-OH (53 mg, 0.28 mmol), EDC1 (53 mg, 0.28 mmol), and DMAP (7 mg, 0.06 mmol). The reaction mixture was stirred at 0°C for 1 hour and diluted with 3 volumes of water while stirring. The resulting precipitate was filtered off and washed with water. The material was then purified by silica gel chromatography eluting with 20%-40% EtOAc/CH2Cl2 gradient. The pure fractions were combined to provide 60 mg of (S,Z)- (4-((7-(cyclopropylamino)-5-(3-(methylsulfonamido)phenyl)pyrazolo[l ,5-a]pyrimidin-3- yl)methylene)-2,5-dioxoimidazolidin-l-yl)methyl 2-(tert-butoxycarbonylamino)propanoate. (LCMS (ES): >95% pure, m/z 655 [M+H]+.
Example 120. Synthesis of (S.ZV(4-((7-(cvclopropylaminoV5-(3- (methylsulfonamido)phenvnpyrazolo l ,5-a1pyrimidin-3-yl")methylene -2,5-dioxoimidazolidin-l - vPmethyl 2-aminopropanoate trifluoromethyl acetate
Figure imgf000307_0001
To (S,Z)-(4-((7-(cyclopropylamino)-5-(3-(methylsulfonamido)phenyl)pyrazolo[l ,5- a]pyrimidin-3-y l)methy lene)-2,5-dioxoimidazolidin- 1 -y l)methy 1 2-(tert- butoxycarbonylamino)propanoate. (60 mg, 0.09 mmol) was added 2M HCl/ether (6 mL) and the reaction mixture was stirred at rt for 2h. The solid was filtered and washed with ether. The material was further purified by mass-directed LC/MS to provide (S,Z)-(4-((7- (cyclopropylamino)-5-(3-(methylsulfonamido)phenyl)pyrazolo[l ,5-a]pyrimidin-3- yl)methylene)-2,5-dioxoimidazolidin-l-yl)methyl 2-aminopropanoate as the TFA salt. (LCMS (ES): >95% pure, m/z 555 [M+H]+.
Figure imgf000307_0002
Same procedure as Example 1 1. LCMS (ES):>90% pure, m/z 468 [M+H]+. Example 122. Synthesis of (S,ZVf4-((7-(cvclopropylamino")-5-(5-(3-fluoroazetidine-l - carbonvnthiophen-2-vnpyrazolo[l ,5-alpyrimidin-3-vnmethyleneV2,5-dioxoimidazolidin-l - vQmethyl 2-(tert-butoxycarbonylamino')propanoate
Figure imgf000308_0001
Same procedure as Example 12. LCMS (ES):>90% pure, m/z 669 [M+H]+.
Example 123. Synthesis of (S^^-fn- cyclopropylamino S- S-n-fluoroazetidine-l - carbonynthiophen^-ynpyrazolol' l .S-alpyrimidin^-vnmethylene^^.S-dioxoimidazolidin-l - yQmethyl 2-aminopropanoate trifluoromethyl acetate
Figure imgf000308_0002
Same procedure as Example 120. LCMS (ES):>90% pure, m z 569 [M+H]+. Table 15. Biological Activities of Examples 120, 121 , and 123.
Figure imgf000308_0003
Figure imgf000309_0001
Example 124. Synthesis of (Z)-½r/-butyl cyclopropyl(3-(Y2.5-dioxoimidazolidin-4- ylidene^ethylVS-H-fluoro^-methyl-l H-indol-S-yloxylpyrazolori .S-alpyrimidin-?- vPcarbamate
Figure imgf000309_0002
(Z)-Tert-bu\y\ 5-chloro-3-((2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[l ,5- a]pyrimidin-7-yl(cyclopropyl)carbamate (75 mg, 0. 18 mmol) was dissolved in anhydrous DMF (0.6 mL). 4-fluoro-5-hydroxy-2-methylindole (45 mg, 0.27 mmol) and 2C03 (75 mg, 0.54 mmol) were added. After 18 h, H20 (3.5 mL) was added to the reaction and precipitate was collected by filtration and dried under high vacuum (~l mmHg) to provide (Z)-tert-butyl cyclopropyl(3-((2,5-dioxoimidazolidin-4-ylidene)methyl)-5-(4-fluoro-2-methyl-l H-indol-5- yloxy)pyrazolo[l ,5-a]pyrimidin-7-yl)carbamate (83 mg, 85%) as a tan solid. LCMS (ES): >90% pure, m z 548 [M+H]+.
Example 125. Synthesis of (Z e -butyl cyclopropyl(3-((2,5-dioxoimtdazolidin-4- ylidene)methyl)-5-(4-fluoro-2-methyl-l H-indol-5-yloxy)pyrazolori ,5-a1pyrimidin-7- vPcarbamate
Figure imgf000310_0001
(Z)-tert-butyl cyclopropyl(3-((2,5-dioxoimidazolidin-4-ylidene)methyl)-5-(4-fluoro-2- methyl-lH-indol-5-yloxy)pyrazolo[l ,5-a]pyrimidin-7-yl)carbamate (40 mg, 0.073 mmol) was dissolved in dichloromethane (0.5 mL) and trifluroacetic acid (0.5 mL). After 1 h, the reaction was concentrated under a stream of air and the residue was triturated with Et20 (4 mL) and filtered to provide (Z)-tert-buty\ cyclopropyl(3-((2,5-dioxoimidazolidin-4-ylidene)methyl)-5-(4- fluoro-2-methyl-l H-indol-5-yloxy)pyrazolo[l ,5-a]pyrimidin-7-yl)carbamate (23 mg, 72%) as a light brown solid.
Example 126. Synthesis of 7-fcyclopropylaminoV5-(4-methoxyphenylthio)pyrazolo[l ,5- alpyrimidine-3-carbaldehyde
Figure imgf000310_0002
To a suspension of tert-butyl 5-chloro-3-formylpyrazolo [1 , 5-a] pyrimidin-7-yl (cyclopropyl) carbamate (50 mg, 0.15 mmol) in DMF (0.4 mL), was added potassium carbonate (25 mg, 0.15 mmol) followed by 4-methoxy benzene thiol (40 μί, 0.3 mmol). The reaction mixture was stirred at 80°C for 1 hour, then cooled to room temperature and diluted with water. The reaction was then partitioned between ethyl acetate and water, the organic layer was dried under sodium sulfate concentrated and dried under vacuum to provide 7-(cyclopropylamino)-5- (4-methoxyphenylthio)pyrazolo[l ,5-a] pyrimidine-3-carbaldehyde. LCMS (ES): >85% pure, m/z 341 [M+H]+.
Example 127. Synthesis of (ZVS-^-CcvclopropylaminoVS-^-methoxyphenylthio'lpyrazolorKS- alpyrimidin-3-y0methylene midazolidine-2,4-dione
Figure imgf000311_0001
To a suspension of 7-(cyclopropylamino)-5-(4-methoxyphenylthio) pyrazolo [1 , 5-a] pyrimidine-3-carbaldehyde in EtOH ( 1 mL), was added hydantoin (53 mg, 0.5 mmol) and piperidine (52 μί). The reaction mixture was stirred at 80°C for three hours and the resulting yellow precipitate was filtered, diluted with 1 : 1 mixture of MeOH and dichloromethane and prepared by HPLC purification to provide (Z)-5-((7-(cyclopropylamino)-5-(4- methoxyphenylthio) pyrazolo[l ,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione. LCMS (ES): >95% pure, m/z 423 [M+H]+.
Table 16. Biological Activities of Examples 124, 125, and 127.
Figure imgf000311_0002
Biological Test Methods:
Biological Example A
CK2 Assay Method
Modulatory activity of compounds described herein was assessed in vitro in cell-free C 2 assays by the following method.
In a final reaction volume of 50 μΐ, CK2 ααββ (4 ng, 8.5 mU) was incubated with various concentrations of test compounds in DMSO (1 ul, 2% by volume), 20 mM MOPS pH 7.2, 10 mM EGTA, 0.15 M NaCl, 10 mM DTT, 0.002% Brij-35, 200 μΜ RRRDDDSDDD, 10 mM MgAcetate, ATP 15 uM and 0.33% (by volume) ([γ-33Ρ]ΑΤΡ: Stock 1 mCi/Ι ΟΟμΙ; 3000 Ci/mmol (Perkin Elmer)). Reactions were maintained for 40 min at 23 °C. The reactions were quenched with 100 ul of 0.75% phosphoric acid, then transferred to and filtered through a phosphocellulose filter plate (Millipore, MSPH-N6B-50). After washing each well 4 times with 0.75% Phosphoric acid, scintillation fluid (20 uL) was added to each well and the residual radioactivity was measured using a Wallac luminescence counter.
Biological Example B
Cell Proliferation Modulatory Activity
A representative cell-proliferation assay protocol using Alamar Blue dye (stored at 4°C, use 20 ul per well) is described hereafter.
96-well plate setup and compound treatment
a. Split and trypsinize cells.
b. Count cells using hemocytometer.
c. Plate 4,000-5,000 cells per well in 100 μΐ of medium and seed into a 96-well plate according to the following plate layout. Add cell culture medium only to wells B 10 to B 12. Wells B l to B9 have cells but no compound added.
1 2 3 4 5 6 7 8 9 10 1 1 12
A EMPTY
Medium
B NO COMPOUND ADDED
Only
C Ι ΟηΜ Ι ΟΟηΜ l uM l OuM Control
D Ι ΟηΜ Ι ΟΟηΜ l uM l OuM Compl
E Ι ΟηΜ Ι ΟΟηΜ l uM l OuM Comp2
F 1'OnM Ι ΟΟηΜ l uM l OuM Comp3
G Ι ΟηΜ Ι ΟΟηΜ l uM l OuM Comp4
H EMPTY d. Add 100 μΐ of 2X drug dilution to each well in a concentration shown in the plate layout above. At the same time, add 100 μΐ of media into the control wells (wells B I O to B l 2). Total volume is 200 μΐ/well.
e. Incubate four (4) days at 37°C, 5% CO2 in a humidified incubator.
,f. Add 20μ1 Alamar Blue reagent to each well.
g. Incubate for four (4) hours at 37°C, 5% C02 in a humidified incubator.
h. Record fluorescence at an excitation wavelength of 544 nm and emission wavelength of 590 nm using a microplate reader.
In the assays, cells are cultured with a test compound for approximately four days, the dye is then added to the cells and fluorescence of non-reduced dye is detected after approximately four hours. Different types of cells can be utilized in the assays (e.g., HCT- 1 16 human colorectal carcinoma cells, PC-3 human prostatic cancer cells, MDA-MB231 human breast cancer cells, K-562 human chronic myelogenous leukemia (CML) cells, MiaPaca human pancreatic carcinoma cells, MV-4 human acute myeloid leukemia cells, and BxPC3 human pancreatic adenocarcinoma cells).
Compounds of the present invention, such as the Examples and compounds listed in various Tables, were tested in these in vitro and cellular assays and have shown desirable ' biological activities. Citation of the above patents, patent applications, publications and documents is not an admission that any of the foregoing is pertinent prior art, nor does it constitute any admission as to the contents or date of these publications or documents. Furthermore, the contents of the patents, patent applications, publications and documents cited herein are incorporated by reference in their entirety for all purposes to the same extent as each and everyone of them is incorporated by references specifically.
Modifications may be made to the foregoing without departing from the basic aspects of the invention. Although the invention has been described in substantial detail with reference to one or more specific embodiments, those of ordinary skill in the art will recognize that changes may be made to the embodiments specifically disclosed in this application, and yet these modifications and improvements are within the scope and spirit of the invention. The invention illustratively described herein suitably may be practiced in the absence of any element(s) not specifically disclosed herein. Thus, for example, in each instance herein any of the terms "comprising", "consisting essentially of, and "consisting of may be replaced with either of the other two terms. Thus, the terms and expressions which have been employed are used as terms of description and not of limitation, equivalents of the features shown and described, or portions thereof, are not excluded, and it is recognized that various modifications are possible within the scope of the invention.

Claims

We claim:
1. A compound of Formula (I):
Figure imgf000315_0001
or a pharmaceutically acceptable salt, solvate, and/or prodrug thereof;
wherein:
Z4 is N or CR5;
R5 is halo, -CN, -R,-OR, -S(0)nR, -COOR, -CONR2, -NR2, optionally substituted aryl, or optionally substituted heteroaryl;
R2, R3 and R4 are each independently H or optionally substituted CI -CIO alkyl;
X is O, S, or NR6;
Y is O or S or NR10;
R6 and R10 are each independently selected from H, CN, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C1-C4 alkoxy, optionally substituted C6-C10 aryl, optionally substituted heteroaryl, and R7R8;
Z is O or S;
L is a covalent bond, -NR7-, -0-, -S(0)n-, -(CR7R8)m-NR9-, -(CR7R8)m-0-, or -(CR7R8)m-
S(0)„-;
W is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, or optionally substituted carbocyclyl;
where each R7 and R8 and R9 is independently selected from H, optionally substituted CI -CIO alkyl, optionally substituted heteroalkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl;
or R 7 and R 8 , taken together on a single carbon atom or on adjacent connected carbon atoms of (CR7R8)m whether alone or as part of another group, form a 3 to 8 membered carbocyclic ring or heterocyclic ring;
or R7 and R8, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 to 10 membered heterocyclic or heteroaryl ring that optionally contains one or more additional heteroatom selected from N, O and S as a ring member;
each m is independently 1, 2, 3 or 4;
G is NR1AR1B, OR1A, or SR1A;
R1A is H or optionally substituted CI -CIO alkyl;
R1B is H, optionally substituted CI -CIO alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, or optionally substituted heteroarylalkyl; each R is independently H or optionally substituted C1-C4 alkyl; or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member; and
each n is independently 0, 1 , or 2.
2. The compound of claim 1, wherein Z4 is N or CH.
3. The compound of claim 1 or 2, wherein R3 and R4 are both H.
4. The compound of any one of claims 1 to 3, wherein R2 is H, -CH3, halo, -OCH3, or -CF3.
5. The compound of any one of claims 1 to 4, wherein Y is O or S.
6. The compound of any one of claims 1 to 5, wherein Z is O.
7. The compound of any one of claims 1 to 6, wherein X is selected from the group consisting of NH, O, and S.
8. The compound of any one of claims 1 to 7, wherein
L is a covalent bond; and
W is optionally substituted aryl, optionally substituted heteroaryl, or optionally substituted heterocyclyl.
9. The compound of any one of claims 1 to 7, wherein
L is -NR7-, -0-, -S(0)„-, -(CR7R8)m-NR9-, -(CR7R8)m-0-, or -(CR7R8)m-S(0)n-;
R 7 and R 8 and R 9 are each independently H or CI -CIO alkyl; and
W is optionally substituted aryl or optionally substituted carbocyclyl.
10. The compound of any one of claims 1 to 7, wherein
W-L- is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, (optionally substituted aryl)-NR7-, (optionally substituted heteroaryl)- NR7-, (optionally substituted aryl)-CH2-NR7-, (optionally substituted heteroaryl)-CH2-NR7-, (optionally substituted aryl)-0-, (optionally substituted heteroaryl)-0-, (optionally substituted aryl)-S-, (optionally substituted heteroaryl)-S-, (optionally substituted aryl)-CH2-0-, or (optionally substituted heteroaryl)-CH2-0-; and
R7 is each independently H or CI -CIO alkyl.
11. The compound of claim 1 , wherein,
Z4 is N or CR5;
R5 is -R, -CN, -OR, -Ar, COOR, -NR2;
Z and Y are both O;
R2 and R4 are both H;
R3 is H, optionally substituted CI -CIO alkyl, or optionally substituted CI -CIO heteroalkyl;
X is NH;
R1B is optionally substituted carbocyclylalkyl; L is a covalent bond, -NR7-, or -CH2-NR7-;
R7 is H or Cl-C4 alkyl; and
W is optionally substituted aryl or optionally substituted heteroaryl.
12. The compound of claim 11, wherein
W is substituted aryl or substituted heteroaryl wherein the one or more substituents are selected from the group consisting of optionally substituted CI -CIO alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted CI -CIO heteroalkyl, -OR, halo, -S(0)R, -S(0)2R, -CN, -NR2, -SR, -N02, -C(0)R, -C(0)OR, -NHC(0)R, -NRC(0)R, - C(0)NR2, -CF3, optionally substituted aryl, and optionally substituted heteroaryl; and
each R is independently selected from H and and optionally substituted C1-C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member, and wherein Ar is optionally substituted aryl or optionally substituted heteroaryl.
13. The compound of claim 12, wherein
W is substituted aryl or substituted heteroaryl wherein the one or more substitutents are selected from the group consisting of phenyl, thiophene, pyridine, pyrimidine, pyrazine pyridazine, pyrrole, furan, thiofuran, oxazole, isooxazole, thiazole, quinoline, isoquinoline, indole, and pyrazole, wherein each substitutent is optionally substituted with one or more substitutents selected from the group consisting of optionally substituted CI -CIO alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted Cl- C10 heteroalkyl, -OR, halo, -S(0)R, -S(0)2R, -CN, -NR2, -SR, -N02, -C(0)R, -C(0)OR, - NHC(0)R, -NRC(0)R, -C(0)NR2, -CF3, optionally substituted aryl, and optionally substituted heteroaryl; and
each R is independently selected from H and and optionally substituted C1-C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member, and wherein Ar is optionally substituted aryl or optionally substituted heteroaryl.
14. The compound of claim 12, wherein
W is optionally substituted aryl or optionally substituted heteroaryl wherein the one or more substitutents are selected from the group consisting of aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine, tetrahydrofuran, pyran, thiopyran, thiomorpholine, thiomorpholine S-oxide, thiomorpholine ^-dioxide, oxazoline, tetrahydrothiophene, piperidine, tetrahydropyran, thiane, imidazolidine, oxazolidine, thiazolidine, dioxolane, dithiolane, piperazine, oxazine, dithiane, morpholine, and dioxane, wherein each substitutent is optionally substituted with one or more substitutents selected from the group consisting of optionally substituted CI -CIO alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted C1-C10 heteroalkyl, -OR, halo, -S(0)R, -S(0)2R, -CN, -NR2, -SR, -N02, - C(0)R, -C(0)OR, -NHC(0)R, -NRC(0)R, -C(0)NR2, -CF3, optionally substituted aryl, and optionally substituted heteroaryl; and
each R is independently selected from H and and optionally substituted C1-C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member, and wherein Ar is optionally substituted aryl or optionally substituted heteroaryl.
15. The compound of claim 12, wherein W is optionally substituted aryl or optionally substituted heteroaryl wherein the one or more substitutents are selected from the group consisting of CI, Br, F, -OCF3, -CF3, -CH3, -C2H5, -C3H7, -OCH20-, OCH2CH20-, -SCH3, - OCH3, -NHCOCH3, -NH2, -COCH3, -N02, -0(CH2)i_6-COOR, and -S02CH3.
16. The compound of any one of claims 12 to 15, wherein W is an optionally substituted phenyl or an optionally substituted pyridine.
17. The compound of claim 1 , having a structural Formula (II):
Figure imgf000320_0001
wherein:
p is 0, 1 , 2 or 3;
Z4 is N or CR5;
R5 is H or Cl-C4 alkyl;
X is NH or S;
R3 is H, CI -CIO alkyl, or CI -CIO heteroalkyl;
R7 is H or Cl-C4 alkyl;
Ar is aryl or heteroaryl which is optionally substituted by one or more substituents selected from the group consisting of optionally substituted CI -CIO alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted C2-C10 alkenyl, optionally substituted C2-C10 alkynyl, -NR2; -OR, S(0)nR; halo, -CN, -N02, -C(0)OR, - C(0)NR2, or -C(0)R;
n is 0, 1 , or 2;
each R is independently H or optionally substituted C1-C4 alkyl; or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member; and
R1B is selected from H, optionally substituted CI -CIO alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, or optionally substituted heteroarylalkyl.
18. The compound of claim 17, wherein Ar is optionally substituted phenyl or pyridinyl.
19. The compound of claim 17 or 18, wherein p is 0 or 1.
The com ound of claim 17, having structural Formula (Ila), (lib), (lie) or (lid)
Figure imgf000321_0001
wherein:
Z4 is N or CR5;
R5 is H or Cl-C4 alkyl;
X is NH, or S;
R3 is H, CI -CIO alkyl or CI -CIO heteroalkyl;
R7 is H or Cl-C4 alkyl;
q is 0, 1 , 2, 3, 4, or 5;
t is 0, 1, 2, 3, or 4;
A is optionally substituted CI -CIO alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted C2-C10 alkenyl, optionally substituted C2-C10 alkynyl, -NR2; -OR, S(0)nR; halo, -CN, -N02, -C(0)OR, -C(0)NR2, or -C(0)R; n is 0, 1 , or 2;
each R is independently H or optionally substituted C1-C4 alkyl; or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member; and
R1B is cyclopropyl or cyclopropylmethylene.
21. The compound of claim 20, wherein A is selected from the group consisting of phenyl, thiophene, pyridine, pyrmidine, pyrazine pyridazine, pyrrole, furan, thiofuran, oxazole isooxazole, thiazole, quinoline isoquinoline, indole, and pyrazole, each of which is optionally substituted with one or more substitutents selected from the group consisting of optionally substituted CI -CIO alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted C1-C10 heteroalkyl, -OR, halo, -S(0)R, -S(0)2R, -CN, -NR2, -SR, -N02, - C(0)R, -C(0)OR, -NHC(0)R, -NRC(0)R, -C(0)NR2, -CF3, optionally substituted aryl, and optionally substituted heteroaryl; and
each R is independently selected from H and and optionally substituted C1-C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member, and wherein Ar is optionally substituted aryl or optionally substituted heteroaryl.
22. The compound of claim 20, wherein A is selected from the group consisting of aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine, tetrahydro furan, pyran, thiopyran, thiomorpholine, thiomorpholine S-oxide, thiomorpholine ^-dioxide, oxazoline,
tetrahydrothiophene, piperidine, tetrahydropyran, thiane, imidazolidine, oxazolidine,
thiazolidine, dioxolane, dithiolane, piperazine, oxazine, dithiane, morpholine, and dioxane, each of which is optionally substituted with one or more substitutents selected from the group consisting of optionally substituted CI -CIO alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted C1-C10 heteroalkyl, -OR, halo, -S(0)R, -S(0)2R,
-CN, -NR2, -SR, -N02, -C(0)R, -C(0)OR, -NHC(0)R, -NRC(0)R, -C(0)NR2, -CF3, optionally substituted aryl, and optionally substituted heteroaryl; and each R is independently selected from H and and optionally substituted C1-C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member, and wherein Ar is optionally substituted aryl or optionally substituted heteroaryl.
23. The compound of claim 20, wherein A is selected from the group consisting of CI, Br, F, -OCF3, -CF3, -CH3, -C2H5, -C3H7, -OCH2O-, OCH2CH2O-, -SCH3, -OCH3, -NHCOCH3, -NH2, - COCH3, -NO2, -0(CH2)i_6-COOR, and -S02CH3.
24. The compound of claim 1 , having a structural Formula (III):
Figure imgf000323_0001
wherein:
Z4 is N or CR5;
R5 is halo, -CN, -R,-OR, -S(0)nR, -COOR, -CONR2, -NR2, optionally substituted aryl, or optionally substituted heteroaryl;
R2, R3 and R4 are each independently is H or optionally substituted CI -CIO alkyl;
X is O, S, or NR6;
Y is O, S, or NR10;
R6 and R10 are each independently selected from H, CN, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C1-C4 alkoxy, optionally substituted C6-C10 aryl, and optionally substituted heteroaryl;
Z is O or S; hCy is an optionally substituted 5 to 10 membered heterocyclic or heteroaryl ring that contains one or more heteroatom selected from N, O and S as a ring member;
G is NR1AR1B, OR1A, or SR1A;
R1A is H or optionally substituted CI -CIO alkyl;
R1B is selected from H, optionally substituted CI -CIO alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, or optionally substituted heteroarylalkyl;
each R is independently H or optionally substituted C1-C4 alkyl; or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member; and
n is 0, 1 , or 2.
25. The compound of claim 24, wherein hCy is selected from the group consisting of thiophene, pyridine, pyrmidine, pyrazine pyridazine, pyrrole, furan, thiofuran, oxazole isooxazole, thiazole, quinoline isoquinoline, indole, and pyrazole,, each of which is optionally substituted with one or more substitutents selected from the group consisting of optionally substituted CI -CIO alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted C1-C10 heteroalkyl, -OR, halo, -S(0)R, -S(0)2R, -CN, -NR2, -SR, -N02, - C(0)R, -C(0)OR, -NHC(0)R, -NRC(0)R, -C(0)NR2, -CF3, optionally substituted aryl, and optionally substituted heteroaryl; and
each R is independently selected from H and and optionally substituted C1-C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member, and wherein Ar is optionally substituted aryl or optionally substituted heteroaryl.
The compound of claim 24, having a structural Formula (Ilia)
Figure imgf000325_0001
wherein:
each A is independently optionally substituted CI -CIO alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted C2-C10 alkenyl, optionally substituted C2-C10 alkynyl, -NR2; -OR, S(0)nR; halo, -CN, -N02, -C(0)OR, - C(0)NR2, or -C(0)R;
c is 0, 1, 2, or 3;
n is 0, 1 , or 2; and
each R is independently H or optionally substituted C1-C4 alkyl; or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member.
27. The compound of claim 26, wherein:
Z4 is CR5;
R2 and R4 are H;
Z and Y are O;
X is NH or S;
G is NR1AR1B;
R1A is H or methyl;
R1B is optionally substituted CI -CIO alkyl, optionally substituted CI -CIO cylcoalkyl, optionally substituted heterocyclyl, optionally substituted CI -CIO alkylether, optionally substituted CI -CIO alkylaminoalkyl, optionally substituted CI -CIO alkoxy alkyl, optionally substituted heterocyclylalkyl, alkylaryl, aralkyl, alkylheteroaryl, heteroarylalkyl, or optionally substituted aryl.
28. The compound of claim 26 or 27, wherein R1B is cyclopropyl, cyclopropylmethylene, cyclobutyl, cyclohexyl, optionally substituted morpholinylalkyl, or optionally substituted phenyl.
29. The compound of any one of claims 26 to 28, wherein A is -CN, C1-C4 alkyl, -C(0)OR, or -C(0)NR2.
30. The compound of claim 24, wherein hCy is selected from the group consisting of aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine, tetrahydrofuran, pyran, thiopyran, thiomorpholine, thiomorpholine S-oxide, thiomorpholine ^-dioxide, oxazoline, tetrahydrothiophene, piperidine, tetrahydropyran, thiane, imidazolidine, oxazolidine,
thiazolidine, dioxolane, dithiolane, piperazine, oxazine, dithiane, morpholine, and dioxane, each of which is optionally substituted with one or more substitutents selected from the group consisting of optionally substituted CI -CIO alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted CI -CIO heteroalkyl, -OR, halo, -S(0)R, -S(0)2R, -CN, -NR2, -SR, -N02, -C(0)R, -C(0)OR, -NHC(0)R, -NRC(0)R, -C(0)NR2, -CF3, optionally substituted aryl, and optionally substituted heteroaryl; and
each R is independently selected from H and and optionally substituted C1-C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member, and wherein Ar is optionally substituted aryl or optionally substituted heteroaryl.
31. The compound of claim 24, having a structural Formula (Illb):
Figure imgf000327_0001
wherein:
D is N or CH;
a and b are independently 1 , 2, or 3;
Z4 is N or CR5;
R5 is halo, -CN, -R,-OR, -S(0)nR, -COOR, -CONR2, -NR2, optionally substituted aryl, or optionally substituted heteroaryl;
2 3 4
R RJ and R" are each independently selected from H and optionally substituted CI -CIO alkyl;
X is O, S, or NR6;
Y is O or S or NR10;
Z is O or S;
R6 and R10 are each independently selected from H, CN, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C1-C4 alkoxy, optionally substituted C6-C10 aryl, and optionally substituted heteroaryl;
L is optionally substituted CI -CIO alkyl, optionally substituted CI -CIO heteroalkyl, optionally substituted C2-C10 alkenyl, optionally substituted C2-C10 alkynyl, optionally substituted aryl, optionally substituted heteroaryl, -C(O)-, -S(0)n-, or -CR2-,
E is -OR, -NR, optionally substituted CI -CIO alkyl, optionally substituted CI -CIO heteroalkyl, optionally substituted aryl, or optionally substituted heteroaryl;
R1B is H, optionally substituted CI -CIO alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, or optionally substituted heteroarylalkyl; and
each R is independently selected from H and optionally substituted C1-C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member; and
n is 0, 1 , or 2.
32. The compound of claim 31 , wherein
Z4 is N or CH;
R2, R3, and R4 are H;
Z and Y are O;
X is NH; and
R1B is cyclopropyl or cyclopropylmethylene.
33. The compound of claim 31 or 32, wherein a and b are both 2.
34. The compound of any of claims 31 to 33, wherein
L2 is -C(O)- or -S(0)2-; and
E is optionally substituted aryl or optionally substituted heteroaryl.
35. The compound of any of claims 31 to 33, wherein
L2 is -CH2-; and
E is optionally substituted C1-C4 alkyl, optionally substituted aryl, or optionally substituted heteroaryl.
36. The compound of claim 34 or 35, wherein the optionally substituted aryl is optionally substituted phenyl, and the optionally substituted heteroaryl is optionally substituted pyridyl.
37. The compound of claim 34 or 35, wherein the optionally substituted aryl or the optionally substituted heteroaryl is independently optionally substituted by one or more substituents selected from the group consisting of optionally substituted CI -CIO alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted CI -CIO heteroalkyl, -OR, halo, -S(0)R, -S(0)2R, -CN, -NR2, -SR, -N02, -C(0)R, -C(0)OR, -NHC(0)R, -NRC(0)R, - C(0)NR2, optionally substituted aryl, and optionally substituted heteroaryl; and
each R is independently selected from H and and optionally substituted C1-C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member.
38. compound of claim 1 having a structural Formula (IV)
Figure imgf000329_0001
wherein:
L1 is O or S;
L3 is a covalent bond or an optionally substituted C 1 -C4 alkylene;
Z4 each independently represent N or CR5;
R5 is halo, -CN, -R,-OR, -S(0)nR, -COOR, -CONR2, -NR2, optionally substituted aryl, or optionally substituted heteroaryl;
each R is independently selected from H and optionally substituted C1-C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member; n is 0, 1 , or 2;
2 3 4
R RJ and R are each independently selected from H and optionally substituted CI -CIO alkyl;
X is O, S, or NR6;
Y is O or S or R10;
R6 and R10 are each independently selected from H, CN, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C1-C4 alkoxy, optionally substituted C6-C10 aryl, and optionally substituted heteroaryl;
Z is O or S;
Ar is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, or optionally substituted heterocyclyl; and
R1B is selected from H, optionally substituted CI -CIO alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl.
39. The compound of claim 38, wherein:
Z4 is CH;
R2, R3, and R4 are H;
Z and Y are O;
X is NH;
R1 is cyclopropyl;
L is a covalent bond or an optionally substituted C 1 -C4 alkylene; and
Ar is optionally substituted aryl or heteroaryl.
40. The compound of claim 38 or 39, wherein L3 is a covalent bond or methylene.
41. The compound of any one of claims 38 to 40, wherein Ar is optionally substituted phenyl, optionally substituted pyridyl, or optionally substituted indolyl.
42. compound of claim 1 having a structural Formula (V)
Figure imgf000331_0001
wherein:
Z4 is N or CR5;
R5 is halo, -CN, -R,-OR, -S(0)nR, -COOR, -CONR2, -NR2, optionally substituted aryl, or optionally substituted heteroaryl;
R2, R3 and R4 are each independently selected from H and optionally substituted CI -CIO alkyl;
X is O, S, or NR6;
Y is O or S or NR10;
Z is O or S;
R6 and R10 are each independently selected from H, CN, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C1-C4 alkoxy, optionally substituted C6-C10 aryl, and optionally substituted heteroaryl;
cCy is a carbocyclic ring which is optionally further substituted;
L4 is -S(0)-NR-, -S(0)2-NR-, -S(0)-0-, -S(0)2-0-, -C(0)-NR-, -C(0)-0-, -NR-S(O)-, - NR-S(0)2-, -O-S(O)-, -0-S(0)2-, -NR-C(O)-, or -O-C(O)-;
Ar is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, or optionally substituted heterocyclyl;
R7 is H, or optionally substituted C1-C4 alkyl; R1 is selected from H, optionally substituted CI -CIO alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; and
each R is independently selected from H and optionally substituted C1-C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member; and
n is 0, 1 , or 2;
43. The compound of claim 42, wherein:
Z4 is N;
R2, R3, and R4 are H;
Z and Y are O;
X is NH;
R1 is cyclopropyl;
L4 is -S(0)-NR-, -S(0)2-NR-, -S(0)-0-, -S(0)2-0-, -C(0)-NR-, -C(0)-0-;
Ar is optionally substituted aryl or optionally substituted heteroaryl;
R7 is H; and
cCy is a cyclohexyl ring.
44. The compound of claim 42 or 43, wherein Ar-L4- is Ar-S(0)2-NR-.
45. The compound of any one of claims 42 to 44, wherein Ar is optionally substituted phenyl.
46. The compound of claim 1 , having a structural Formula (VI):
Figure imgf000333_0001
wherein:
Z4 is N or CR5;
R5 is halo, -CN, -R, -OR, -S(0)nR, -C(0)OR, -C(0)NR2, -NR2 or -Ar;
each R is independently selected from H and optionally substituted C1-C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member,
n is 0, 1 , or 2;
2 3 4
R RJ and R" are each independently selected from H and optionally substituted CI -CIO alkyl;
X is O, S, or NR6;
Y is O, S, or NR10;
R6 and R10 are each independently selected from H, CN, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C1-C4 alkoxy, optionally substituted C6-C10 aryl, and optionally substituted heteroaryl;
Z is O or S;
Ar is an optionally substituted aryl; and
R1B is selected from H, optionally substituted CI -CIO alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, or optionally substituted heteroarylalkyl.
47. The compound of claim 46 having a structural Formula (Via):
Figure imgf000334_0001
wherein
each A is independently optionally substituted CI -CIO alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted C2-C10 alkenyl, optionally substituted C2-C10 alkynyl, -NR2; -OR, -S(0)nR, -NR-S(0)nR, halo, -CN, -N02, - C(0)OR, -0-C(0)-R, -C(0)NR2, -NR-C(0)R, or -C(0)R;
c is 0, 1, 2, 3, 4, or 5;
each n is independently 0, 1 , or 2; and
each R is independently H or optionally substituted C1-C4 alkyl; or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that is optionally contains one or more additional heteroatom selected from N, O, and S as a ring member.
48. The compound of claim 46 or 47, wherein R3 is -alkylene-0-C(0)-R3a, wherein R3a is optionally substituted C1-C4 alkyl, optionally substituted C1-C4 heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, or optionally substituted heterocyclyl.
49. The compound of claim 48, wherein -C(0)-R3a is a carboxyl terminus of an amino acid.
50. The compound of claim 49, wherein the amino acid is L-Alanine.
51. The compound of claim 1 , which is selected from the species disclosed in the specification.
52. A pharmaceutical composition comprising
a compound of any one of claims 1 to 51 ; and
at least one pharmaceutically acceptable excipient.
53. The pharmaceutical composition of claim 52, further comprising one or more additional therapeutic agent.
54. The pharmaceutical composition of claim 53, wherein the one or more additional therapeutic agent is an anticancer agent.
55. A method for modulating casein kinase 2 activity and/or Pim kinase activity in a cell comprising contacting the cell with a compound of any one of claims 1 to 51 , or a
pharmaceutically acceptable salt, solvate, and/or prodrug thereof.
56. A method of treating a condition or disease associated with casein kinase 2 activity and/or Pim kinase activity in a patient comprising administering to the patient a therapeutically effective amount of the compound of any one of claims 1 to 51 , or a pharmaceutically acceptable salt, solvate, and/or prodrug thereof.
57. The method of claim 56, wherein the condition or disease is selected from a group consisting of a cancer, a vascular disorder, a inflammation, a pathogenic infection, a
immunological disorder, a neurodegenerative disorder, and a combination thereof.
58. The method of claim 57, wherein the cancer is of the colorectum, breast, lung, liver, pancreas, lymph node, colon, prostate, brain, head and neck, skin, liver, kidney, blood, ovary or heart.
59. The method of claim 56, comprising administering to the patient the compound of any one of claims 1 to 51, or a pharmaceutically acceptable salt, solvate, and/or prodrug thereof, in combination with one or more additional therapeutic agent.
60. The method of claim 59, wherein the one or more additional therapeutic agent is an anticancer agent.
61. A method for inhibiting cell proliferation, which comprises contacting cells with the compound of any one of claims 1 to 51 , or a pharmaceutically acceptable salt, solvate, and/or prodrug thereof, in an amount effective to inhibit proliferation of the cells.
62. The method of claim 61 , wherein the cells are in a cancer cell line or in a tumor in a subject.
63. The method of claim 62, wherein the cancer cell line is a breast cancer, prostate cancer, pancreatic cancer, lung cancer, hematopoietic cancer, colorectal cancer, skin cancer, ovary cancer cell line.
64. A method for inhibiting angiogenesis in a subject, which comprises administering to the subject the compound of any one of claims 1 to 51, or a pharmaceutically acceptable salt, solvate, and/or prodrug thereof, in an amount effective to inhibit the angiogenesis.
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