WO2014033630A1

WO2014033630A1 - Novel aminothiazole carboxamides as kinase inhibitors

Info

Publication number: WO2014033630A1
Application number: PCT/IB2013/058033
Authority: WO
Inventors: Matthew Burger; Yu Ding; Wooseok Han; Gisele Nishiguchi; Alice Rico; Robert Lowell Simmons; Huw Tanner; Lifeng Wan
Original assignee: Novartis Ag
Priority date: 2012-08-31
Filing date: 2013-08-27
Publication date: 2014-03-06

Abstract

The present invention provides a compound of formula (I), as described herein, and pharmaceutically acceptable salts, enantiomers, rotamers, tautomers, or racemates thereof. Also provided are methods of treating a disease or condition mediated by PIM kinase using the compounds of Formula I, and pharmaceutical compositions comprising such compounds.

Description

NOVEL AMINOTHIAZOLE CARBOXAMIDES AS KINASE INHIBITORS

FIELD OF THE INVENTION

The present invention relates to new compounds and compositions of the new compounds together with pharmaceutically acceptable carriers, and uses of the new compounds, either alone or in combination with at least one additional therapeutic agent, in the prophylaxis or treatment of cancer and other cellular proliferation disorders.

BACKGROUND

Infection with the Moloney retrovirus and genome integration in the host cell genome results in development of lymphomas in mice. Provirus Integration of Moloney Kinase (PIM-Kinase) was identified as one of the frequent proto-oncogenes capable of being transcriptionally activated by this retrovirus integration event (Cuypers HT et al, "Murine leukemia virus-induced T-cell lymphomagenesis: integration of pro viruses in a distinct chromosomal region," Cell 37(1): 141-50 (1984); Selten G, et al, "Proviral activation of the putative oncogene Pim-1 in MuLV induced T-cell lymphomas" EMBO J 4(7): 1793-8 (1985)), thus establishing a correlation between over-expression of this kinase and its oncogenic potential. Sequence homology analysis demonstrated that there are three highly homologous Pim-Kinases (Piml, 2 & 3), Piml being the proto-oncogene originally identified by retrovirus integration. Furthermore, transgenic mice over- expressing Piml or Pim2 show increased incidence of T-cell lymphomas (Breuer M et al., "Very high frequency of lymphoma induction by a chemical carcinogen in pim-1 transgenic mice" Nature 340(6228):61-3 (1989)), while over-expression in conjunction with c-myc is associated with incidence of B-cell lymphomas (Verbeek S et al., "Mice bearing the E mu-myc and E mu-pim-1 transgenes develop pre-B-cell leukemia prenatally" Mol Cell Biol 11(2): 1176-9 (1991)). Thus, these animal models establish a strong correlation between Pirn over-expression and oncogenesis in hematopoietic malignancies. In addition to these animal models, Pirn over-expression has been reported in many human malignancies, particularly in hematopoietic and in prostate cancer. Furthermore, mutational activation of several well known oncogenes in hematopoietic malignancies is thought to exert its effects at least in part through Pim(s). For example, targeted down-regulation of Pirn expression impairs survival of hematopoietic cells transformed by Flt3 and BCR/ABL (Adam et al. 2006). Piml, 2 & 3 are Serine/Threonine kinases that normally function in survival and proliferation of hematopoietic cells in response to growth factors and cytokines. Substrates for Pim kinases include regulators of apoptosis such as the Bcl-2 family member BAD. The effects of Pim(s) in these regulators are consistent with a role in protection from apoptosis and promotion of cell proliferation and growth. Thus, over- expression of Pim(s) in cancer is thought to play a role in promoting survival and proliferation of cancer cells and, therefore, their inhibitions should be an effective way of treating cancers in which they are over-expressed. In fact several reports indicate that knocking down expression of Pim(s) with siRNA results in inhibition of proliferation and cell death (Dai JM, et al., "Antisense oligodeoxynucleotides targeting the serine/threonine kinase Pim-2 inhibited proliferation of DU-145 cells," Acta Pharmacol Sin 26(3):364-8 (2005); Fujii et al. 2005; Li et al. 2006). Thus, inhibitors to Piml, 2 and 3 would be useful in the treatment of these malignancies.

In addition to a potential role in cancer treatment and myeloproliferative diseases, such inhibitors could be useful to control expansion of immune cells in other pathologic condition such as autoimmune diseases, allergic reactions and in organ transplantation rejection syndromes. Recent reports demonstrate that Pim kinase inhibitors show activity in animal models of inflammation and autoimmune diseases. See JE Robinson "Targeting the Pim Kinase Pathway for Treatment of Autoimmune and Inflammatory Diseases," for the Second Annual Conference on Anti-Inflammatories: Small Molecule Approaches," San Diego, CA (Conf. April 2011; Abstract published earlier on-line).

A continuing need exists for compounds that inhibit the proliferation of capillaries, inhibit the growth of tumors, treat cancer, modulate cell cycle arrest, and/or inhibit molecules such as Piml, Pim2 and Pim3, and pharmaceutical formulations and medicaments that contain such compounds. A need also exists for methods of administering such compounds, pharmaceutical formulations, and medicaments to patients or subjects in need thereof. The present invention addresses such needs.

Earlier patent applications have described compounds that inhibit Pirns and function as anticancer therapeutics, see, e.g., WO2012/004217, WO2010/026124, WO 2008/106692 and WO2011/124580, and as treatment for inflammatory conditions such as Crohn's disease, inflammatory bowel disease, rheumatoid arthritis, and chronic inflammatory diseases, see e.g., WO 2008/022164. The present invention provides novel compounds that inhibit activity of one or more Pirns, preferably two or more Pirns, more preferably Piml, Pim2 and Pim3, at nanomolar levels (e.g., IC-50 under 50 nM) and exhibit distinctive characteristics that may provide improved therapeutic effects and pharmacokinetic properties, such as reduced drug-drug interactions associated with inhibition of cytochrome oxidases, relative to compounds previously disclosed. Compounds of the invention contain novel substitution combinations on one or more rings that provide these distinctive properties and are suitable for treating Pim-related conditions such as those described herein.

SUMMARY OF THE INVENTION

The invention provides compounds of Formula (I) that inhibit one or more Pirn kinases:

(I)

wherein:

represents a pyridine ring or N-(Ci-C₄ alkyl)-pyrazole ring;

W is H or NH₂;

Z is CH or N;

n is 1 or 2;

ft¹ is H, OH or OMe;

provided that when R¹ is H, R² is also H;

R³ is H or Ci-C₄ alkyl; R⁴ is selected from R*, -OR*, -OCH₂CH₂OR*, -CH₂OR*, and a 4-6 membered cyclic ether optionally substituted with OH, OMe, or F, wherein each R* is independently C 1-C4 alkyl,

provided that R⁴ is not -OMe when Z is N;

or a pharmaceutically acceptable salt thereof.

Additional embodiments of these compounds and pharmaceutical compositions and uses for these compounds and compositions are described below.

These compounds are inhibitors of Pirn kinases as further discussed herein. These compounds and their pharmaceutically acceptable salts, and pharmaceutical compositions containing these compounds and salts, are useful for therapeutic methods such as treatment of cancers and autoimmune disorders that are caused by or exacerbated by excessive levels of Pirn kinase activity. DETAILED DESCRIPTION OF EMBODIMENTS OF THE INVENTION

"PIM inhibitor" is used herein to refer to a compound that exhibits an IC₅₀ with respect to PIM Kinase activity of no more than about 100 μΜ and more typically not more than about 5 μΜ, as measured in the PIM depletion assays described herein below for at least one of Piml, Pim2 and Pim3. Preferred compounds have on IC₅₀ below about 1 micromolar on at least one Pirn, and generally have an IC₅₀ below 100 nM on each of Piml, Pim2 and Pim3.

The phrase "alkyl" refers to hydrocarbon groups that do not contain heteroatoms, i.e., they consist of carbon atoms and hydrogen atoms. Thus the phrase includes straight chain alkyl groups such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl and the like. The phrase also includes branched chain isomers of straight chain alkyl groups, including but not limited to, the following which are provided by way of example: -

CH(CH₃)₂, -CH(CH₃)(CH₂CH₃), -CH(CH₂CH₃)₂, -C(CH₃)₃, -C(CH₂CH₃)₃, -CH₂CH(CH₃) ₂, -CH₂CH(CH₃)(CH₂CH₃), -CH₂CH(CH₂CH₃)₂, -CH₂C(CH₃)₃, -CH₂C(CH₂CH₃)₃, -CH(C ¾)-

CH(CH₃)(CH₂CH₃), -CH₂CH₂CH(CH₃)₂, -CH₂CH₂CH(CH₃)(CH₂CH₃), -CH₂CH₂CH(CH ₂CH₃)₂, -CH₂CH₂C(CH₃)₃, -CH₂CH₂C(CH₂CH₃)₃, -CH(CH₃)CH₂_

CH(CH₃)₂, -CH(CH₃)CH(CH₃)CH(CH₃)₂, -CH(CH₂CH₃)CH(CH₃)CH(CH₃)(CH₂CH₃), and others. Thus the term 'alkyl' includes primary alkyl groups, secondary alkyl groups, and tertiary alkyl groups. Alkyl groups are described herein according to the number of carbon atoms they contain, e.g., an alkyl group containing up to six carbon atoms is described as a CI -6 or C_1-6, or C1-C6 alkyl. Typical alkyl groups include straight and branched chain alkyl groups having 1 to 12 carbon atoms, preferably 1-6 carbon atoms. The term 'lower alkyl' or "loweralkyl" and similar terms refer to alkyl groups containing up to 6 carbon atoms.

The term "alkenyl" refers to alkyl groups as defined above, wherein there is at least one carbon-carbon double bond, i.e., wherein two adjacent carbon atoms are attached by a double bond. The term "alkynyl" refers to alkyl groups wherein two adjacent carbon atoms are attached by a triple bond. Typical alkenyl and alkynyl groups contain 2-12 carbon atoms, preferably 2-6 carbon atoms. Lower alkenyl or lower alkynyl refers to groups having up to 6 carbon atoms. An alkenyl or alkynyl group may contain more than one unsaturated bond, and may include both double and triple bonds, but of course their bonding is consistent with well-known valence limitations.

The term 'alkoxy" refers to -OR, wherein R is alkyl.

As used herein, the term "halogen" or "halo" refers to chloro, bromo, fluoro and iodo groups. Typical halo substituents are F and/or CI. "Haloalkyl" refers to an alkyl radical substituted with one or more halogen atoms, typically 1-3 halogen atoms. The term "haloalkyl" thus includes monohalo alkyl, dihalo alkyl, trihalo alkyl, perhaloalkyl, and the like.

"Amino" refers herein to the group -NH₂. The term "alkylamino" refers herein to the group -NRR where R and R are each independently selected from hydrogen or a lower alkyl, provided -NRR' is not -NH₂. The term "arylamino" refers herein to the group -NRR where R is aryl and R' is hydrogen, a lower alkyl, or an aryl. The term "aralkylamino" refers herein to the group -NRR where R is a lower aralkyl and R is hydrogen, a loweralkyl, an aryl, or a loweraralkyl.

The term "alkoxyalkyl" refers to the group -alki-0-alk₂ where alki is an alkyl linking group, and alk₂ is alkyl, e.g., a group such as -0-(CH₂)₂-0-CH₃. The term "loweralkoxyalkyl" refers to an alkoxyalkyl where alki is loweralkyl and alk₂ is loweralkyl. The term "aryloxyalkyl" refers to the group -alkyl-O-aryl, where -alkyl- is a Ci_i2 straight or branched chain alkyl linking group, preferably C_1-6. The term "aralkoxyalkyl" refers to the group -alkyl-O-aralkyl, where aralkyl is preferably a loweraralkyl.

The term "aminocarbonyl" refers herein to the group -C(0)-NH₂ . "Substituted aminocarbonyl" refers herein to the group -C(0)-NRR' where R is loweralkyl and R' is hydrogen or a loweralkyl. In some embodiments, R and R, together with the N atom attached to them may be taken together to form a "heterocycloalkylcarbonyl" group. The term "arylaminocarbonyl" refers herein to the group -C(0)-NRR where R is an aryl and R is hydrogen, loweralkyl or aryl. "aralkylaminocarbonyl" refers herein to the group - C(0)-NRR' where R is loweraralkyl and R is hydrogen, loweralkyl, aryl, or loweraralkyl.

"Aminosulfonyl" refers herein to the group -S(0)₂-NH₂. "Substituted aminosulfonyl" refers herein to the group -S(0)2-NRR' where R is loweralkyl and R' is hydrogen or a loweralkyl. The term "aralkylaminosulfonylaryl" refers herein to the group -aryl-S(0)₂-NH-aralkyl, where the aralkyl is loweraralkyl.

"Carbonyl" refers to the divalent group -C(O)-. "Carboxy" refers to-C(=0)-OH.

"Alkoxycarbonyl" refers to ester -C(=0)-OR wherein R is optionally substituted lower alkyl. "Loweralkoxycarbonyl" refers to ester -C(=0)-OR wherein R is optionally substituted lower loweralkyl. "Cycloalkyloxycarbonyl" refers to -C(=0)-OR wherein R is optionally substituted C3-C8 cycloalkyl.

"Cycloalkyl" refers to a mono- di- or poly-cyclic, carbocyclic alkyl substituent in which all ring atoms are carbon. Typical cycloalkyl groups have from 3 to 8 backbone (i.e., ring) atoms. When used in connection with cycloalkyl substituents, the term "polycyclic" refers herein to fused and non-fused alkyl cyclic structures, including spirocyclic ring systems. The term "partially unsaturated cycloalkyl", "partially saturated cycloalkyl", and "cycloalkenyl" all refer to a cycloalkyl group wherein there is at least one unsaturated carbon-carbon bond in the ring, i.e., wherein two adjacent ring atoms are connected by a double bond or a triple bond. Such rings typically contain 1 or 2 double bonds for 5-6 membered rings, and 1-2 double bonds or one triple bond for 7-8 membered rings. Illustrative examples include cyclohexenyl, cyclooctynyl, cyclopropenyl, cyclobutenyl, cyclohexadienyl, and the like.

The term "heterocycloalkyl" refers herein to cycloalkyl substituents that have from 1 to 5, and more typically from 1 to 3 heteroatoms as ring members in place of carbon atoms. Preferably, heterocycloalkyl or "heterocyclyl" groups contain one or two heteroatoms as ring members, typically only one heteroatom for 3-5 membered rings and 1-2 heteroatoms for 6-8 membered rings. Suitable heteroatoms employed in heterocyclic groups of the present invention are nitrogen, oxygen, and sulfur. Representative heterocycloalkyl moieties include, for example, pyrrolidinyl, tetrahydrofuranyl, oxirane, oxetane, oxepane, thiirane, thietane, azetidine, morpholino, piperazinyl, piperidinyl and the like.

The terms "substituted heterocycle", "heterocyclic group" or "heterocycle" as used herein refers to any 3- or 4-membered ring containing a heteroatom selected from nitrogen, oxygen, and sulfur or a 5- or 6-membered ring containing from one to three heteroatoms, preferably 1-2 heteroatoms, selected from the group consisting of nitrogen, oxygen, or sulfur; wherein the 5 -membered ring has 0-2 double bonds and the 6-membered ring has 0-3 double bonds; wherein the nitrogen and sulfur atom maybe optionally oxidized; wherein the nitrogen and sulfur heteroatoms may be optionally quaternized; and including any bicyclic group in which any of the above heterocyclic rings is fused to a benzene ring or another 5- or 6-membered heterocyclic ring or heteroaryl as described herein. Preferred heterocycles include, for example: diazapinyl, pyrrolinyl, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, N-methyl piperazinyl, azetidinyl, N-methylazetidinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, and oxiranyl. The heterocyclic groups may be attached at various positions as will be apparent to those having skill in the organic and medicinal chemistry arts in conjunction with the disclosure herein.

Heterocyclic moieties can be unsubstituted or they can be substituted with one or more substituents independently selected from hydroxy, halo, oxo (C=0), alkylimino (RN=, wherein R is a loweralkyl or loweralkoxy group), amino, alkylamino, dialkylamino, acylaminoalkyl, alkoxy, thioalkoxy, lower alkoxyalkoxy, loweralkyl, cycloalkyl or haloalkyl. Typically, substituted heterocyclic groups will have up to four substituent groups.

The term "cyclic ether" as used herein refers to a 3-7 membered ring, unless otherwise specified, containing one oxygen atom (O) as a ring member. Where the cyclic ether is "optionally substituted" it can be substituted at any carbon atom with a group suitable as a substituent for a heterocyclic group, typically up to three substituents selected from lower alkyl, lower alkoxy, halo, hydroxy, amino, -C(0)-lower alkyl, and - C(0)-lower alkoxy. In preferred embodiments, halo, hydroxy and lower alkoxy are not attached to the carbon atoms of the ring that are bonded directly to the oxygen atom in the cyclic ether ring. Specific examples include oxirane, oxetane (e.g., 3-oxetane), tetrahydrofuran (including 2-tetrahydrofuranyl and 3-tetrahydrofuranyl), tetrahydropyran (e.g., 4-tetrahydropyranyl), and oxepane.

"Aryl" refers to monocyclic and polycyclic aromatic groups having from 5 to 14 backbone carbon or hetero atoms, and includes both carbocyclic aryl groups and heteroaromatic aryl groups. Carbocyclic aryl groups are aryl groups in which all ring atoms in the aromatic ring are carbon, typically including phenyl and naphthyl. Exemplary aryl moieties employed as substituents in compounds of the present invention include phenyl, pyridyl, pyrimidinyl, thiazolyl, indolyl, imidazolyl, oxadiazolyl, tetrazolyl, pyrazinyl, triazolyl, thiophenyl, furanyl, quinolinyl, purinyl, naphthyl, benzothiazolyl, benzopyridyl, and benzimidazolyl, and the like. When used in connection with aryl substituents, the term "polycyclic aryl" refers herein to fused and non-fused cyclic structures in which at least one cyclic structure is aromatic, such as, for example, benzodioxozolo (which has a heterocyclic structure fused to a phenyl group, naphthyl, and the like. Where "aryl" is used, the group is preferably a carbocyclic group; the term "heteroaryl" is used for aryl groups when ones containing one or more heteroatoms are preferred.

The term "heteroaryl" refers herein to aryl groups having from 1 to 4 heteroatoms as ring atoms in an aromatic ring with the remainder of the ring atoms being carbon atoms, in a 5-14 atom aromatic ring system that can be monocyclic or polycyclic. Monocyclic heteroaryl rings are typically 5-6 atoms in size. Exemplary heteroaryl moieties employed as substituents in compounds of the present invention include pyridyl, pyrimidinyl, thiazolyl, indolyl, imidazolyl, oxadiazolyl, tetrazolyl, pyrazinyl, triazolyl, thiophenyl, furanyl, quinolinyl, purinyl, benzothiazolyl, benzopyridyl, and benzimidazolyl, and the like.

"Aralkyl" or "arylalkyl" refers to an aryl group connected to a structure through an alkylene linking group, e.g., a structure such as -(CH₂)i_4-Ar, where Ar represents an aryl group. "Lower aralkyl" or similar terms indicate that the alkyl linking group has up to 6 carbon atoms.

"Optionally substituted" or "substituted" refers to the replacement of one or more hydrogen atoms with a non-hydrogen group. Alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups described herein may be substituted or unsubstituted. Suitable substitution groups include, for example, hydroxy, nitro, amino, imino, cyano, halo, thio, sulfonyl, thioamido, amidino, imidino, oxo, oxamidino, methoxamidino, imidino, guanidino, sulfonamido, carboxyl, formyl, loweralkyl, haloloweralkyl, loweralkylamino, haloloweralkylamino, lower alkoxy, lower haloalkoxy, lower alkoxyalkyl, alkylcarbonyl, aminocarbonyl, arylcarbonyl, aralkylcarbonyl, heteroarylcarbonyl, heteroaralkylcarbonyl, alkylthio, aminoalkyl, cyanoalkyl, aryl and the like, provided that oxo, imidino or other divalent substitution groups are not placed on aryl or heteroaryl rings due to the well known valence limitations of such rings. In preferred embodiments, unless otherwise specified, optional substituents for alkyl, alkenyl, alkynyl, cycloalkyl, and heterocycloalkyl groups are 1-3 groups selected from halo, hydroxy, amino, cyano, lower alkoxy, lower alkylsulfonyl, oxy, carboxy, and lower alkoxy carbonyl. In preferred embodiments, unless otherwise specified, optional substituents for aryl and heteroaryl groups are 1-3 groups selected from halo, hydroxy, amino, cyano, lower alkyl, lower alkoxy, lower alkylsulfonyl, carboxy, and lower alkoxy carbonyl.

The substitution group can itself be substituted where valence permits, i.e., where the substitution group contains at least one CH, NH or OH having a hydrogen atom that can be replaced. The group substituted onto the substitution group can be carboxyl, halo (on carbon only); nitro, amino, cyano, hydroxy, loweralkyl, loweralkoxy, C(0)R, - OC(0)R, -OC(0)OR, -NRCOR, -CONR₂, -NRCOOR, -C(S)NR₂, -NRC(S)R, - OC(0)NR₂, , -SR, -SO₃H, -S0₂R or C3-8 cycloalkyl or 3-8 membered heterocycloalkyl, where each R is independently selected from hydrogen, lower haloalkyl, lower alkoxyalkyl, and loweralkyl, and where two R on the same atom or on directly connected atoms can be linked together to form a 5-6 membered heterocyclic ring. Unless indicated as optionally substituted, these substitution groups are typically unsubstituted.

When a substituted substituent includes a straight chain group, the substitution can occur either within the chain (e.g., 2-hydroxypropyl, 2-aminobutyl, and the like) or at the chain terminus (e.g., 2-hydroxyethyl, 3-cyanopropyl, and the like). Substituted substituents can be straight chain, branched or cyclic arrangements of covalently bonded carbon or heteroatoms.

It is understood that the above definitions are not intended to include impermissible substitution patterns (e.g., methyl substituted with five fluoro groups or a halogen atom substituted with another halogen atom). Such impermissible substitution patterns are well known to the skilled artisan.

"Syn" as used herein has its ordinary meaning, and is used in connection with Formula I to indicate that the specified groups are attached to sp³ hybridized (tetrahedral) carbon centers and extend out from one face of the cyclohexyl or piperidinyl ring, i.e., those groups all project toward the 'alpha' face of the ring, or they all project toward the 'beta' face of the ring. This is thus used as a convenient way to define the relative orientations of two or more groups on a ring, without limiting the compounds to a specific absolute chiral configuration. This reflects the fact that the compounds of the invention have such groups in a specific relative orientation, but are not limited to either enantiomer of that specific relative orientation. Accordingly, unless described as optically active, such compounds may be racemic, but also include each of the two enantiomers having the specified relative stereochemistry. In some embodiments, the compounds of the invention are optically active form as further described herein, and in preferred embodiments of the invention, the compounds are obtained and used in optically active form. Preferably, the enantiomer having greater potency as an inhibitor of at least two of Piml, Pim2 and Pim3 is selected.

It will also be apparent to those skilled in the art that the compounds of the invention, as well as the pharmaceutically acceptable salts, esters, metabolites and prodrugs of any of them, may be subject to tautomerization and may therefore exist in various tautomeric forms wherein a proton of one atom of a molecule shifts to another atom and the chemical bonds between the atoms of the molecules are consequently rearranged. See, e.g., March, Advanced Organic Chemistry: Reactions, Mechanisms and Structures, Fourth Edition, John Wiley & Sons, pages 69-74 (1992). As used herein, the term "tautomer" refers to the compounds produced by the proton shift, and it should be understood that all tautomeric forms, insofar as they may exist, are included within the invention.

The compounds of the invention comprise one or more asymmetrically substituted carbon atoms. Such asymmetrically substituted carbon atoms can result in the compounds of the invention existing in enantiomers, diastereomers, and other stereoisomeric forms that may be defined, in terms of absolute stereochemistry, such as in (R)- or (S)- forms. The compounds of the invention are sometimes depicted herein as single enantiomers, and are intended to encompass the specific configuration depicted and the enantiomer of that specific configuration (the mirror image isomer of the depicted configuration), unless otherwise specified— e.g., where a structure is labeled 'chiral', it represents the specified absolute stereochemistry as a single substantially pure (i.e., at least about 95% pure) enantiomer. The depicted structures herein describe the relative stereochemistry of the compounds where two or more chiral centers, but the invention is not limited to the depicted enantiomer's absolute stereochemistry unless otherwise stated. The invention includes both enantiomers, each of which will exhibit Pim inhibition, even though one enantiomer will be more potent than the other. In some instances, compounds of the invention have been synthesized in racemic form and separated into individual isomers by chiral chromatography or similar conventional methods, and the analytical data about the two enantiomers do not provide definitive information about absolute stereochemical configuration. In such cases, the absolute stereochemistry of the most active enantiomer has been identified based on correlation with similar compounds of known absolute stereochemistry, rather than by a definitive physical method such as X- ray crystallography. Therefore, in certain embodiments, the preferred enantiomer of a compound described herein is the specific isomer depicted or its opposite enantiomer, whichever has the lower IC-50 for Pim kinase inhibition using the assay methods described herein, i.e., the enantiomer that is more potent as a Pim inhibitor for at least two of Pim 1, Pim2, and Pim3.

The terms "S" and "R" configuration, as used herein, are as defined by the IUPAC

1974 RECOMMENDATIONS FOR SECTION E, FUNDAMENTAL STEREOCHEMISTRY, Pure Appl. Chem. 45: 13-30 (1976). The terms a and β are employed for ring positions of cyclic compounds. The a-side of the reference plane is that side on which the preferred substituent lies at the lower numbered position. Those substituents lying on the opposite side of the reference plane are assigned β descriptor. It should be noted that this usage differs from that for cyclic stereoparents, in which "a" means "below the plane" and denotes absolute configuration. The terms a and β configuration, as used herein, are as defined by the CHEMICAL ABSTRACTS INDEX GUIDE -APPENDIX IV (1987) paragraph 203.

As used herein, the term "pharmaceutically acceptable salts" refers to the nontoxic acid or base addition salts of the compounds of Formulas I, II, etc., wherein the compound acquires a positive or negative charge as a result of adding or removing a proton; the salt then includes a counterion of opposite charge from the compound itself, and the counterion is preferably one suitable for pharmaceutical administration under the conditions where the compound would be used. These salts can be prepared in situ during the final isolation and purification of the compounds of Formula I or II, or by separately reacting the base or acid functions with a suitable organic or inorganic acid or base, respectively. Representative salts include but are not limited to the following: acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate, digluconate, cyclopentanepropionate, dodecylsulfate, ethanesulfonate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, fumarate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, pamoate, pectinate, persulfate, 3-phenylproionate, picrate, pivalate, propionate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate and undecanoate.

Also, a basic nitrogen-containing group in compounds of the invention can be quaternized with such agents as loweralkyl halides, such as methyl, ethyl, propyl, and butyl chloride, bromides, and iodides; dialkyl sulfates like dimethyl, diethyl, dibutyl, and diamyl sulfates, long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides, aralkyl halides like benzyl and phenethyl bromides, and others. Water or oil-soluble or dispersible products are thereby obtained. These quaternized ammonium salts when paired with a pharmaceutically acceptable anion can also serve as pharmaceutically acceptable salts.

Examples of acids which may be employed to form pharmaceutically acceptable acid addition salts include such inorganic acids as hydrochloric acid, sulfuric acid and phosphoric acid and such organic acids as oxalic acid, maleic acid, methanesulfonic acid, succinic acid and citric acid. Basic addition salts can be prepared in situ during the final isolation and purification of the compounds of formula (I), or separately by reacting carboxylic acid moieties with a suitable base such as the hydroxide, carbonate or bicarbonate of a pharmaceutically acceptable metal cation or with ammonia, or an organic primary, secondary or tertiary amine. Counterions for pharmaceutically acceptable salts include, but are not limited to, cations based on the alkali and alkaline earth metals, such as sodium, lithium, potassium, calcium, magnesium, aluminum salts and the like, as well as nontoxic ammonium, quaternary ammonium, and amine cations, including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like. Other representative organic amines useful for the formation of base addition salts include diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine and the like.

As used herein, the term "pharmaceutically acceptable ester" refers to esters, which hydrolyze in vivo and include those that break down readily in the human body to leave the parent compound or a salt thereof. Suitable ester groups include, for example, those derived from pharmaceutically acceptable aliphatic carboxylic acids, particularly alkanoic, alkenoic, cycloalkanoic and alkanedioic acids, in which each alkyl or alkenyl moiety advantageously has not more than 6 carbon atoms. Examples of particular pharmaceutically acceptable esters include formates, acetates, propionates, maleates, lactates, hydroxyacetates, butyrates, acrylates and ethylsuccinates.

The term "pharmaceutically acceptable prodrugs" as used herein refers to those prodrugs of the compounds of the present invention which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended use, as well as the zwitterionic forms, where possible, of the compounds of the invention. The term "prodrug" refers to compounds that are rapidly transformed in vivo to yield the parent compound of the above formula, for example by hydrolysis in blood. A thorough discussion is provided in T. Higuchi and V. Stella, PRO-DRUGS AS NOVEL DELIVERY SYSTEMS, Vol. 14 of the A.C.S. Symposium Series, and in Edward B. Roche, ed., BIOREVERSIBLE CARRIERS IN DRUG DESIGN, American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated herein by reference.

Any formula given herein is also intended to represent unlabeled forms as well as isotopically labeled forms of the compounds, lsotopically labeled compounds have structures depicted by the formulas given herein except that one or more atoms are replaced by an atom having a selected atomic mass or mass number. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen,

2 3H 11 13 carbon, nitrogen, oxygen, phosphorous, fluorine, and chlorine, such as H, , C, C, ¹⁴C, ¹⁵N, ¹⁸F ³¹P, ³²P, ³⁵S, ³⁶C1, ¹²⁵I respectively. The invention includes various isotopically labeled compounds as defined herein, for example those into which radioactive isotopes, such as ³H and ¹⁴C, or those into which non-radioactive isotopes, such as ²H and ¹³C are present. Such isotopically labeled compounds are useful in metabolic studies (with ¹⁴C), reaction kinetic studies (with, for example ²H or ³H), detection or imaging techniques, such as positron emission tomography (PET) or single- photon emission computed tomography (SPECT) including drug or substrate tissue distribution assays, or in radioactive treatment of patients. In particular, an 18F or labeled compound may be particularly desirable for PET or SPECT studies. Isotopically-labeled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying Examples and Preparations using an appropriate isotopically-labeled reagents in place of the non-labeled reagent previously employed.

Further, substitution with heavier isotopes, particularly deuterium (i.e., ²H or D) may afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements or an improvement in therapeutic index. It is understood that deuterium in this context is regarded as a substituent of a compound of the formula (I). The concentration of such a heavier isotope, specifically deuterium, may be defined by the isotopic enrichment factor. The term "isotopic enrichment factor" as used herein means the ratio between the isotopic abundance and the natural abundance of a specified isotope. If a substituent in a compound of this invention is denoted deuterium, such compound has an isotopic enrichment factor for each designated deuterium atom of at least 3500 (52.5% deuterium incorporation at each designated deuterium atom), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90%) deuterium incorporation), at least 6333.3 (95%> deuterium incorporation), at least 6466.7 (97%) deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5%> deuterium incorporation).

Pharmaceutically acceptable solvates in accordance with the invention include those wherein the solvent of crystallization may be isotopically substituted, e.g. D₂0, d⁶- acetone, d⁶-DMSO.

Compounds of the invention, i.e. compounds of formula (I) that contain groups capable of acting as donors and/or acceptors for hydrogen bonds may be capable of forming co-crystals with suitable co-crystal formers. These co-crystals may be prepared from compounds of formula (I) by known co-crystal forming procedures. Such procedures include grinding, heating, co-subliming, co-melting, or contacting in solution compounds of formula (I) with the co-crystal former under crystallization conditions and isolating co-crystals thereby formed. Suitable co-crystal formers include those described in WO 2004/078163. Hence the invention further provides co-crystals comprising a compound of formula (I).

The following enumerated embodiments are representative of certain aspects of the invention:

1. A compound of Formula (I) :

₄ alkyl)-pyrazole ring;

W is H or NH₂;

Z is CH or N;

n is 1 or 2;

R¹ is H, OH or OMe;

R² is H or Me,

provided that when R¹ is H, R² is also H;

R³ is H or Ci-C₄ alkyl;

R⁴ is selected from R*, -OR*, -OCH₂CH₂OR*, -CH₂OR*, and a 4-6 membered cyclic ether optionally substituted with OH, OMe, or F, wherein each R* is independently C₁-C4 alkyl,

provided that R⁴ is not -OMe when Z is N; or a pharmaceutically acceptable salt thereof. In preferred embodiments, W is

NH₂.

2. The compound of embodiment 1, wherein R³ is H.

3. The compound of embodiment 1, wherein R³ is Me.

4. The compound of any of embodiments 1-3, wherein R² is H.

5. The compound of any of embodiments 1-3, wherein R² is Me.

6. The compound of any of embodiments 1-3, wherein R¹ is H.

7. The compound of any of embodiments 1-5, wherein R¹ is OH.

8. The compound of any of the preceding embodiments, wherein Z is CH.

9. The compound of any of the preceding embodiments, which is of the formula IA:

The compound of any one of embodiments 1-7, wherein Z is N.

The compound of embodiment 10, which is of the formula IB:

12. The compound of any of the preceding embodiments, wherein n is 1.

The compound of embodiment 10 or 11, wherein

The compound of embodiment 9 or 11 , wherein represents

pyrazole of the formula where R is methyl, ethyl or isopropyl.

15. The compound of embodiment 14, wherein R^N is methyl.

The compound of any one of embodiments 1-12, wherein represents

a pyridine of the formula

17. The compound of any of the preceding embodiments, wherein R is a

tetrahydropyranyl group of the formula:

wherein R^T is H, OH, OMe, or F. In certain embodiments, it is selected from H, OH and F.

18. The compound of any of embodiments 1-16, wherein R is an oxetanyl group of

wherein R is H, OH, OMe, or F. In certain embodiments, it is selected from H, OH and F.

19. The compound of any of embodiments 1-16, wherein R is OMe, OEt, or OPr.

20. The compound of any of embodiments 1-16, wherein R⁴ is -CH₂OMe or -CH₂OEt 21. The compound of embodiment 1, which is selected from the compounds in Table 1. Each compound and any subset of the compounds in Table 1 represent preferred embodiments of the invention. 22. A pharmaceutical composition comprising a compound of any of the preceding embodiments and at least one pharmaceutically acceptable excipient.

23. The pharmaceutical composition of embodiment 22, further comprising an additional therapeutic agent.

24. The pharmaceutical composition of embodiment 23, wherein the additional therapeutic agent is selected from irinotecan, topotecan, gemcitabine, 5-fluorouracil, cytarabine, daunorubicin, PI3 Kinase inhibitors, mTOR inhibitors, DNA synthesis inhibitors, leucovorin, carboplatin, cisplatin, taxanes, tezacitabine, cyclophosphamide, vinca alkaloids, imatinib, anthracyclines, rituximab, and trastuzumab.

25. A method to treat a condition caused or exacerbated by excessive Pirn kinase activity, wherein the method comprises administering to a subject in need thereof an effective amount of a compound of any of embodiments 1-21. In some embodiments, the subject has been diagnosed with a condition caused by Pirn kinase.

26. The method of embodiment 25, wherein the condition is a cancer.

27. The method of embodiment 26, wherein the cancer is selected from carcinoma of the lungs, pancreas, thyroid, ovary, bladder, breast, prostate, or colon, melanoma, myeloid leukemia, multiple myeloma, erythroleukemia, villous colon adenoma, and osteosarcoma; or the autoimmune disorder is selected from Crohn's disease, inflammatory bowel disease, rheumatoid arthritis, and chronic inflammatory diseases. 28. A compound according to any of embodiments 1-21 for use in therapy.

29. Use of a compound according to any one of embodiments 1-21 for the preparation of a medicament. The medicament may be to treat a cancer, such as a cancer selected from carcinoma of the lungs, pancreas, thyroid, ovaries, bladder, breast, prostate or colon, melanoma, myeloid leukemia, multiple myeloma, erythro leukemia, villous colon adenoma, and osteosarcoma. 31. The compound according to embodiment 30, wherein the cancer is selected from carcinoma of the lungs, pancreas, thyroid, ovaries, bladder, breast, prostate or colon, melanoma, myeloid leukemia, multiple myeloma, erythro leukemia, villous colon adenoma, and osteosarcoma. 32. The compound of embodiment 31 , wherein the condition is an autoimmune disorder.

33. A method of treating a disease or condition mediated by PIM kinase, comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of embodiments 1-25, or a pharmaceutically acceptable salt thereof.

34. The method of embodiment 33, wherein the disease is selected from carcinoma of the lungs, pancreas, thyroid, ovaries, bladder, breast, prostate or colon, melanoma, myeloid leukemia, multiple myeloma, erythro leukemia, villous colon adenoma, and osteosarcoma; or the disease is an autoimmune disorder.

35. The method of embodiment 34, wherein the disease is an autoimmune disorder. 36. The method of embodiment 35, wherein the autoimmune disorder is selected from Crohn's disease, inflammatory bowel disease, rheumatoid arthritis, and chronic inflammatory diseases.

In the compounds of the invention, Ring A can be pyridyl or certain N- alkylpyrazoles; preferred embodiments of Ring A include:

where R is methyl, ethyl or isopropyl, especially Me;

where (Z) indicates the position of Ring A that is attached to the ring containing Z.

In the compounds of Formula I, Z can be CH or N; when Z is N, preferred embodiments of the ring containing Z include:

where R¹ is H or OH; R² is H or Me, provided that when R¹ is H, R² is also H; and R³ is H, Me, Et or iPr.

Alternatively, when Z is CH, preferred embodiments of the ring containing Z include:

where R¹ is H or OH; R² is H or Me, provided that when R¹ is H, R² is also H; and R³ is Me, Et or iPr. In the compounds where Z is CH, n is preferably 1, while n is often 2 when Z is N.

The substituent R⁴ can influence in vivo properties such as metabolism and clearance rates as well as drug interactions, particularly in combination with the above embodiments of the ring containing Z. Some of the preferred embodiments of R⁴ in compounds of the invention include: Me, OMe, iPr, -OiPr, -CH₂OEt, -CH₂OiPr, - OCH₂CH₂OMe, and groups comprising optionally substituted cyclic ethers, including these:

In compounds of the invention, R¹ is preferably H or OH, and R² is preferably H. R³ is typically Me; or R³ can be H, especially when R¹ is H and/or n is 2.

These compounds may be used in racemic form, or the individual enantiomers may be used, or mixtures of the enantiomers may be used. Each enantiomer can be used, and preferably the compound to be used is the enantiomer that has greater activity as a Pirn inhibitor.

For purposes of the present invention, a therapeutically effective dose will generally be a total daily dose administered to a host in single or divided doses may be in amounts, for example, of from 0.001 to 1000 mg/kg body weight daily, typically 0.01 to 100 mg/kg per day, and more preferred from 0.1 to 30 mg/kg body weight daily. Generally, daily dosage amounts of 1 to 4000 mg, or from 5 to 3000, or from 10 to 2000 mg, or from 100 to 2000 mg are anticipated for human subjects. Dosage unit compositions may contain such amounts of submultiples thereof to make up the daily dose. The compounds of the present invention may be administered orally, parenterally, sublingually, by aerosolization or inhalation spray, rectally, or topically in dosage unit formulations containing conventional nontoxic pharmaceutically acceptable carriers, adjuvants, and vehicles as desired. Topical administration may also involve the use of transdermal administration such as transdermal patches or ionophoresis devices. The term parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrasternal injection, or infusion techniques. In preferred embodiments, the compound or composition of the invention is administered orally.

Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-propanediol. Among the acceptable vehicles and solvents that may 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 may be employed including synthetic mono- or di-glycerides. In addition, 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 and polyethylene glycols, which are solid 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 may include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound may be admixed with at least one inert diluent such as sucrose lactose or starch. Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., lubricating agents such as magnesium stearate. In the case of capsules, tablets, and pills, the dosage forms may also comprise buffering agents. Tablets and pills can additionally be prepared with enteric coatings.

Liquid dosage forms for oral administration may include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art, such as water. Such compositions may also comprise adjuvants, such as wetting agents, emulsifying and suspending agents, cyclodextrins, and sweetening, flavoring, and perfuming agents.

The compounds of the present invention can also be administered in the form of liposomes. As is known in the art, liposomes are generally derived from phospholipids or other lipid substances. Liposomes are formed by mono- or multi-lamellar hydrated liquid crystals that are dispersed in an aqueous medium. Any non-toxic, physiologically acceptable and metabolizable lipid capable of forming liposomes can be used. The present compositions in liposome form can contain, in addition to a compound of the present invention, stabilizers, preservatives, excipients, and the like. The preferred lipids are the phospholipids and phosphatidyl cholines (lecithins), both natural and synthetic. Methods to form liposomes are known in the art. See, for example, Prescott, Ed., Methods in Cell Biology, Volume XIV, Academic Press, New York, N.W., p. 33 et seq. (1976).

While the compounds of the invention can be administered as the sole active pharmaceutical agent, they can also be used in combination with one or more other agents used in the treatment of cancer. The compounds of the present invention are also useful in combination with known therapeutic agents and anti-cancer agents, and combinations of the presently disclosed compounds with other anti-cancer or chemotherapeutic agents are within the scope of the invention. Examples of such agents can be found in Cancer Principles and Practice of Oncology, V. T. Devita and S. Hellman (editors), 6^th edition (Feb. 15, 2001), Lippincott Williams & Wilkins Publishers. 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. Such anti-cancer agents include, but are not limited to, the following: MEK inhibitors, estrogen receptor modulators, androgen receptor modulators, retinoid receptor modulators, cytotoxic/cytostatic agents, antiproliferative agents, prenyl-protein transferase inhibitors, HMG-CoA reductase inhibitors and other angiogenesis inhibitors, inhibitors of cell proliferation and survival signaling, apoptosis inducing agents and agents that interfere with cell cycle checkpoints. The compounds of the invention are also useful when co- administered with radiation therapy.

Therefore, in one embodiment of the invention, the compounds of the invention are also used in combination with known therapeutic or anticancer agents including, for example, estrogen receptor modulators, androgen receptor modulators, retinoid receptor modulators, cytotoxic agents, antiproliferative agents, prenyl-protein transferase inhibitors, HMG-CoA reductase inhibitors, HIV protease inhibitors, reverse transcriptase inhibitors, and other angiogenesis inhibitors.

In certain presently preferred embodiments of the invention, representative therapeutic agents useful in combination with the compounds of the invention for the treatment of cancer include, for example, MEK inhibitors, irinotecan, topotecan, gemcitabine, 5-fluorouracil, cytarabine, daunorubicin, PI3 Kinase inhibitors, mTOR inhibitors, DNA synthesis inhibitors, leucovorin carboplatin, cisplatin, taxanes, tezacitabine, cyclophosphamide, vinca alkaloids, imatinib (Gleevec), anthracyclines, rituximab, trastuzumab, Revlimid, Velcade, dexamethasone, daunorubicin, cytaribine, clofarabine, Mylotarg, lenalidomide, bortezomib, as well as other cancer

chemotherapeutic agents including targeted therapeutics.

The above compounds to be employed in combination with the compounds of the invention will be used in therapeutic amounts as indicated in the Physicians' Desk Reference (PDR) 47th Edition (1993), which is incorporated herein by reference, or such therapeutically useful amounts as would be known to one of ordinary skill in the art, or provided in prescribing materials such as a drug label for the additional therapeutic agent.

The compounds of the invention and the other anticancer agents can be administered at the recommended maximum clinical dosage or at lower doses. Dosage levels of the active compounds in the compositions of the invention may be varied so as to obtain a desired therapeutic response depending on the route of administration, severity of the disease and the response of the patient. The combination can be administered as separate compositions or as a single dosage form containing both agents. When administered as a combination, the therapeutic agents can be formulated as separate compositions, which are given at the same time or different times, or the therapeutic agents, can be given as a single composition.

In one embodiment, the invention provides a method of inhibiting Piml, Pim2 or Pim3 in a human or animal subject. The method includes administering an effective amount of a compound, or a pharmaceutically acceptable salt thereof, of any of the embodiments of compounds of Formula I or II to a subject in need thereof.

The present invention will be understood more readily by reference to the following examples, which are provided by way of illustration and are not intended to be limiting of the present invention. Synthetic Methods

The compounds of the invention can be obtained through procedures known to those skilled in the art. As shown in Scheme 1, 5-alkyl, 4-hydroxy, 3-aminopiperidines can be prepared and modified to yield 5-alkyl, 4-substituted, 3-aminopiperidinyl pyridine amides VI as follows. Reaction of Garner's aldehyde with (R)-4-benzyl-3- propionyloxazolidin-2-one followed by TBS protection of the resulting alcohol affords compound I. Reduction of the oxazolidinone followed by introduction of the azide group yields intermediate II. Deprotection under acidic conditions reveals the corresponding amino alcohol, which upon protection with the Boc group followed by mesylation of the primary alcohol yields intermediate III. Reduction of the azide affords formation of the piperidine which is subsequently reacted with 4-chloro-3-nitropyridine and following nitro reduction pyridyl aniline IV is obtained. Aniline IV can be coupled with heterocyclic acids, which after silyl group and N-Boc deprotection can afford target amides VI.

Scheme 1

In Scheme 2, synthetic methods to prepare certain aminocyclohexylpyridyl amides

X are depicted. Methyl cyclohexanedione can be converted via the monotriflate to the corresponding cyclohexenoneboronate ester which can undergo palladium mediated carbon bond formation with 4-chloro, 3-nitro pyridine to yield nitropyridine substituted cyclohexenone VII. Ketone reduction followed by dehydration yields a cyclohexadiene which upon epoxidation (via bromohydrin formation and HBr elimination), azide epoxide opening, azide reduction and amine Boc protection yields cyclohexenyl Boc amino alcohol nitro pyridyl compound VIII. Nitro pyridyl VIII can be converted to the trans protected amino hydroxy aniline IX by alcohol protection and alkene and nitro reduction. As described above in Scheme 1 for the preparation of substituted piperidine compounds of the invention, upon amide coupling of the cyclohexyl pyridyl anilines IX to heterocyclic acids and subsequent hydroxyl and amine deprotection, substituted cyclohexyl compounds of the invention X can be prepared. Scheme 2

Alternatively, as shown in Scheme 3, cyclohexanediones can be converted via monotriflates to the corresponding cyclohexenoneboronate esters which can undergo palladium mediated carbon bond formation with 4-chloro, 3-nitro pyridine to yield nitropyridine substituted cyclohexenones XI. Reduction of the enone functionality can yield a cyclohexenol XII, which can undergo Mitsunobu reaction with phthalimide to yield a protected aminocyclohexene XIII. Following nitro and alkene reduction, phthalimide protected aminocyclohexyl pyridyl aniline Va can undergo amide coupling and deprotection, to yield aminocyclohexane amides XIV. The corresponding Boc protected aminocyclohexane pyridyl aniline XV can also be prepared from cyclohexenol XII in the following manner: alcohol protection, alkene and nitro reduction, pyridyl amine Cbz protection, silyl ether deprotection, Dess-Martin oxidation to the cyclohexanone, reductive amination with benzylamine, Cbz and Bn deprotection and primary aliphatic amine Boc protection. Penultimate amide products which contain a heteroaromatic bromo functionality can be further modified by standard modifications to introduce substituted aryls, alkyls and heteroaryls on place of !¾. For example, if R2 is Br, by reaction with boronic acids or organometallic reagents, or conversion to the corresponding boronate ester and reaction with aryl/heteroaryl halides or triflates, a variety of R2 replacements are possible. Scheme 3

XV ^N HCI/dioxane XVI

Referring to the examples that follow, compounds of the preferred embodiments were synthesized using the methods described herein, or other methods, which are known in the art.

The compounds and/or intermediates were characterized by high performance liquid chromatography (HPLC) using a Waters Millenium chromatography system with a 2695 Separation Module (Milford, MA). The analytical columns were reversed phase Phenomenex Luna CI 8 -5 μ, 4.6 x 50 mm, from Alltech (Deerfield, IL). A gradient elution was used (flow 2.5 mL/min), typically starting with 5% acetonitrile/95% water and progressing to 100% acetonitrile over a period of 10 minutes. All solvents contained 0.1%) trif uoroacetic acid (TFA). Compounds were detected by ultraviolet light (UV) absorption at either 220 or 254 nm. HPLC solvents were from Burdick and Jackson (Muskegan, MI), or Fisher Scientific (Pittsburgh, PA).

In some instances, purity was assessed by thin layer chromatography (TLC) using glass or plastic backed silica gel plates, such as, for example, Baker-Flex Silica Gel 1B2-F flexible sheets. TLC results were readily detected visually under ultraviolet light, or by employing well-known iodine vapor and other various staining techniques. Mass spectrometric analysis was performed on one of three LCMS instruments: a Waters System (Alliance HT HPLC and a Micromass ZQ mass spectrometer; Column: Eclipse XDB-C18, 2.1 x 50 mm; gradient: 5-95% (or 35-95%, or 65-95% or 95-95%) acetonitrile in water with 0.05% TFA over a 4 min period; flow rate 0.8 mL/min; molecular weight range 200-1500; cone Voltage 20 V; column temperature 40°C), another Waters System (ACQUITY UPLC system and a ZQ 2000 system; Column: ACQUITY UPLC HSS-C18, 1.8um, 2.1 x 50mm; gradient: 5-95% (or 35-95%, or 65-95% or 95-95%) acetonitrile in water with 0.05% TFA over a 1.3 min period; flow rate 1.2 mL/min; molecular weight range 150-850; cone Voltage 20 V; column temperature 50°C) or a Hewlett Packard System (Series 1100 HPLC; Column: Eclipse XDB-C18, 2.1 x 50 mm; gradient: 5-95% acetonitrile in water with 0.05% TFA over a 4 min period; flow rate 0.8 mL/min; molecular weight range 150-850; cone Voltage 50 V; column temperature 30°C). All masses were reported as those of the protonated parent ions.

Nuclear magnetic resonance (NMR) analysis was performed on some of the compounds with a Varian 400 MHz NMR (Palo Alto, CA). The spectral reference was either TMS or the known chemical shift of the solvent.

Preparative separations are carried out using a Flash 40 chromatography system and KP-Sil, 60A (Biotage, Charlottesville, VA), or by flash column chromatography using silica gel (230-400 mesh) packing material on ISCO or Analogix purification systems, or by HPLC using a Waters 2767 Sample Manager, C-18 reversed phase column, 30X50 mm, flow 75 mL/min. Typical solvents employed for the Flash 40 Biotage, ISCO or Analogixsystem for silica gel column chromatography are dichloromethane, methanol, ethyl acetate, hexane, n-heptanes, acetone, aqueous ammonia (or ammonium hydroxide), and triethyl amine. Typical solvents employed for the reverse phase HPLC are varying concentrations of acetonitrile and water with 0.1% trifluoroacetic acid.

Chiral separation of enantiomeric mixtures was performed by the following analytical and preparative general methods:

Chiral SFC-Analytical Method: Chiral compounds were separated on a Waters

Supercritical Fluid Chromatography (SFC). The separation used a Chiralpak AD (AS, OD, OJ, IC or IA) 4.6x100mm column at 40C temperature at a flow rate of 5 mL/min using an isocratic method. The mobile phase was 15% MeOH (or EtOH or IPA or with 0.1% Diethyl amine): 85% C02. The detection wavelength was 220 nm (or 250nm or Diode Array).

Chiral SFC-Purification Method: Chiral compounds were separated on a Waters Supercritical Fluid Chromatography (SFC). The separation used a Chiralpak AD (AS, OD, OJ, IC or IA) 21x250mm column at 40C temperature at a flow rate of 100 mL/min using an isocratic method. The mobile phase was 15% MeOH (or EtOH or IPA or with 0.1% Diethyl amine): 85% C02. The detection wavelength was 220 nm (or 250nm or Diode Array).

Chiral HPLC-Analytical Method: Chiral compounds were separated on a Waters 2695 HPLC system. The separation used a Chiralpak AD (AS, OD, OJ, IC or IA) 4.6x100mm column at room temperature at a flow rate of 1 mL/min using an isocratic method. The mobile phase was 15% EtOH (or IPA or with 0.1% Diethyl amine): 85% Heptane. The detection wavelength was 220 nm (or 250nm or Diode Array).

Chiral HPLC-Purification Method: Chiral compounds were separated on a Waters 2767 HPLC system. The separation used a Chiralpak AD (AS, OD, OJ, IC or IA) 21x250mm column at room temperature at a flow rate of 20 (or 10 -15) mL/min using an isocratic method. The mobile phase was 15% EtOH (or IPA or with 0.1% Diethyl amine): 85% Heptane. The detection wavelength was 220 nm (or 250nm or Diode Array).

It should be understood that the organic compounds according to the preferred embodiments may exhibit the phenomenon of tautomerism. As the chemical structures within this specification can only represent one of the possible tautomeric forms, it should be understood that the preferred embodiments encompasses any tautomeric form of the drawn structure.

It is understood that the invention is not limited to the embodiments set forth herein for illustration, but embraces all such forms thereof as come within the scope of the above disclosure.

The examples below as well as throughout the application, the following abbreviations have the following meanings. If not defined, the terms have their generally accepted meanings. ABBREVIATIONS

DAST (diethylamino)sulfurtrifluoride

DCM Dichloromethane

DIAD diisopropylazodicarboxylate

DIEA diisopropylethylamine

DMA Dimethylacetamide

DMAP 4-dimethylaminopyridine

DME 1 ,2-dimethoxyethane

DMF N,N-dimethylformamide

DPPF 1 , 1 '-bis(diphenylphosphino)ferrocene

EDC 1 -(3 -Dimethylaminopropyl)-3 -ethylcarbodiimide hydrochloride

EtOAc ethyl acetate

EtOH Ethanol

HOAT Hydroxyazabenzotriazole

K₂C0₃ Potassium carbonate

MeCN Acetonitrile

MgS0₄ Magnesium sulfate

MeOH Methanol

Na₂C0₃ sodium carbonate

NaCl Sodium chloride

NaHC0₃ sodium bicarbonate

NBS N-bromosuccinimide

NMP N-methyl-2-pyrrolidone

Pd₂(dba)₃ Tris(dibenzylideneacetone)dipalladium(0)

Pd(PPh₃)₄ Tetrakis(triphenylphospine)palladium(0)

Pd(dppf)Cl₂- Dichloro-( 1 ,2-bis(diphenylphosphino)ethan)- DCM Palladium(II) - dichloromothethane adduct

RT or rt room temperature

TBDMSC1 tert-butyldimethylsilylchloride ABBREVIATIONS

TEA Triethylamine

THF tetrahydrofuran

EXAMPLES

Synthesis of (R)-tert-butyl 4-((lR,2R)-3-((R)-4-benzyl-2-oxooxazolidin-3-yl)-l-hydroxy- 2-methyl-3-oxopropyl)-2,2-dimethyloxazolidine-3-carboxylate

To a solution of (R)-4-benzyl-3-propionyloxazolidin-2-one (1.0 equiv.) in DCM (0.13 M) was added TiCl₄ (1.0 equiv.) at -40 °C. The mixture was stirred at -40 °C for 10 min (yellow suspension), then DIPEA (2.5 equiv.) was added (dark red solution) and stirred at 0 °C for 20 min. (R)-tert-butyl 4-formyl-2,2-dimethyloxazolidine-3-carboxylate (1.0 equiv.) in DCM (0.5 M) was then added dropwise and the resulting mixture was stirred for 1.5 hours. The reaction was quenched by the addition of aqueous ammonium chloride and the mixture was extracted with ethyl acetate. The organic phase was separated, washed with brine, dried with magnesium sulfate, filtered, and concentrated. The residue was purified via column chromatography eluting with ethyl acetate and hexanes (1 :4) to give (R)-tert-butyl 4-((lR,2R)-3-((R)-4-benzyl-2-oxooxazolidin-3-yl)-l- hydroxy-2-methyl-3-oxopropyl)-2,2-dimethyloxazolidine-3-carboxylate as the major product (5:2) in 58% yield. LC/MS = 363.3 (M+H-Boc), Rt = 1.09 min. Synthesis of flO-tert-butyl 4-(Y lR,2R -3-((R -4-benzyl-2-oxooxazolidin-3-vn-l-(tert- butyldimethylsilyloxy)-2-methyl-3-oxopropyl)-2,2-dimethyloxazolidine-3-carboxylate

To a solution of (R)-tert-butyl 4-((lR,2R)-3-((R)-4-benzyl-2-oxooxazolidin-3-yl)- l-hydroxy-2-methyl-3-oxopropyl)-2,2-dimethyloxazolidine-3-carboxylate (1.0 equiv.) and lutidine (1.8 equiv.) in DCM (0AM) was added TBSOTf (1.4 equiv.) at -40 °C. The reaction mixture was stirred at -40 °C for 2 hours. The solution was diluted with ethyl acetate and washed with sat. NaHC0₃, sat. NaCl, dried with magnesium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography eluting with ethyl acetate and hexanes (1 :4) to give (R)-tert-butyl 4-((lR,2R)-3-((R)-4-benzyl-2- oxooxazolidin-3-yl)-l-(tert-butyldimethylsilyloxy)-2-methyl-3-oxopropyl)-2,2- dimethyloxazolidine-3-carboxylate as the major product (5 :2) in 83% yield. LC/MS = 577.3 (M+H), Rt = 1.33 min (Frac 65%-95% method).

Synthesis of (RV tert-butyl 4-(( 1 R.2SV 1 -(tert-butyldimethylsiryloxyV3 -hydroxy-2- methylpropyl)-2,2-dimethyloxazolidine-3-carboxylate

To a solution of (R)-tert-butyl 4-((lR,2R)-3-((R)-4-benzyl-2-oxooxazolidin-3-yl)- l-(tert-butyldimethylsilyloxy)-2-methyl-3-oxopropyl)-2,2-dimethyloxazolidine-3- carboxylate (1.0 equiv.) and ethanol (3.0 equiv.) in THF (0.09 M) was added LiBH₄ (3.0 equiv.) at -30 °C. The reaction mixture was allowed to warm up to 0 °C and stirred at that temperature for 3 hours. The solution was then diluted with diethyl ether and IN NaOH was added. The resulting mixture was extracted with ethyl acetate, the organic layer was separated, washed with sat. NaCl, dried over magnesium sulfate, filtered, and concentrated. The residue was purified via silica gel column chromatography eluting with ethyl acetate and hexanes (1 :4) to give (R)-tert-butyl 4-((lR,2S)-l-(tert- butyldimethylsilyloxy)-3-hydroxy-2-methylpropyl)-2,2-dimethyloxazolidine-3- carboxylate as the major product (5:2 ratio) in 71% yield. LC/MS = 304.3 (M+H-Boc), Rt = 0.95 min (Frac 65%-95% method). Synthesis of (RVtert-butyl 4-(Y lR,2S -3-azido-l-(tert-butyldimethylsilyloxy -

2-methylpropyl)-2,2-dimethyloxazolidine-3-carboxylate

To a solution of (R)-tert-butyl 4-((lR,2S)-l-(tert-butyldimethylsilyloxy)-3- hydroxy-2-methylpropyl)-2,2-dimethyloxazolidine-3-carboxylate (1.0 equiv.), DIAD (2.0 equiv.), and PPh₃ (2.0 equiv.) in THF (0.18 M) was added DPPA (2.0 equiv., 1M solution in THF). The reaction mixture was stirred at room temperature overnight. Upon removal of the volatiles under vacuo, the residue was purified by silica gel column chromatography eluting with ethyl acetate and hexanes (1 :6) to give (R)-tert-butyl 4- ((lR,2S)-3-azido-l-(tert-butyldimethylsilyloxy)-2-methylpropyl)-2,2-dimethyloxazol- idine-3-carboxylate as the major product (5:2) in 86% yield. LC/MS = 329.3 (M+H- Boc), Rt = 1.40 min (Frac 65%-95% method).

Synthesis of tert-butyl (2R,3R,4S)-5-azido-3-(tert-butyldimethylsilyloxy)- l-hydroxy-4-methylpentan-2-ylcarbamate

To a solution of (R)-tert-butyl 4-((lR,2S)-3-azido-l-(tert-butyldimethylsilyloxy)- 2-methylpropyl)-2,2-dimethyloxazolidine-3-carboxylate (1.0 equiv.) in EtOH (0.1 M) was added PPTS (1.3 equiv.) and the mixture was refluxed for 2 days. The volatiles were removed under vacuo, the residue was dissolved in DCM (0.1 M) and DIEA (1.5 equiv.) and Boc₂0 (1.0 equiv.) were added to the reaction mixture. The solution was stirred for 3 hours at room temperature. The solvents were removed under reduced pressure and the residue was diluted with ethyl acetate, washed with water, aqueous NaHS0₄, aqueous NaHC0₃, sat. NaCl, the organic phase was dried with magnesium sulfate, filtered, and concentrated. The residue was purified via silica gel column chromatography eluting with ethyl acetate and hexanes (1 :3) to give tert-butyl (2R,3R,4S)-5-azido-3-(tert- butyldimethylsilyloxy)-l-hydroxy-4-methylpentan-2-ylcarbamate as the major isomer (5 :2) in 70% yield. LC/MS = 289.3 (M+H-Boc), Rt = 0.76 min (Frac 65%-95% method).

Synthesis of (2R,3R,4S)-5-azido-2-(tert-butoxycarbonylamino)-3-(tert- butyldimethylsilyloxy)-4-methylpentyl methanesulfonate

To a solution of tert-butyl (2R,3R,4S)-5-azido-3-(tert-butyldimethylsilyloxy)-l- hydroxy-4-methylpentan-2-ylcarbamate (1.0 equiv.) in pyridine (0.2 M) was added MsCl (1.3 equiv.) followed by DMAP (catalytic amount) at 0 °C. The mixture was stirred at that temperature for 1 hour. The solution was diluted with ether and ethyl acetate (4:1), washed with aq. NaHSC"4, sat. NaHC0₃, brine, dried over magnesium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography eluting with ethyl acetate and hexanes (1 :3) to give (2R,3R,4S)-5-azido-2-(tert- butoxycarbonylamino)-3-(tert-butyldimethylsilyloxy)-4-methylpentyl methanesulfonate as the major isomer (5:2) in 90% yield. LC/MS = 367.3 (M+H-Boc), Rt = 0.81 min (Frac 65%-95% method).

Synthesis of tert-butyl (3R.4R.5SV4-(tert-butyldimethylsilyloxyV

5 -methylpiperidin-3 -ylcarbamate

A solution of (2R,3R,4S)-5-azido-2-(tert-butoxycarbonylamino)-3-(tert- butyldimethylsilyloxy)-4-methylpentyl methanesulfonate in MeOH (0.09 M) was degassed with nitrogen for 20 min. DIEA (2.5 equiv.) was added, followed by 10%> Pd/C (0.1 equiv.). The reaction mixture was stirred under a hydrogen balloon for 2 hours. The solution was filtered and the filtrate was concentrated under vacuo to afford tert-butyl (3R,4R,5S)-4-(tert-butyldimethylsilyloxy)-5-methylpiperidin-3-ylcarbamate as the major isomer (5:2) in >99% yield. LC/MS = 345.2 (M+H-Boc), Rt = 0.95 and 0.99 min.

Synthesis of tert-butyl (3R.4R.5 S -4-(tert-butyldimethylsilyloxy -5-methyl- 1 -(3- nitropyridin-4-yl)piperidin-3-ylcarbamate

BocHN

To a solution of tert-butyl (3R,4R,5S)-4-(tert-butyldimethylsilyloxy)-5- methylpiperidin-3-ylcarbamate (1.0 equiv.) in i-PrOH (0.09 M) was added DIEA (2.5 equiv.) and 4-chloro-3-nitropyridine (1.5 equiv.). The reaction mixture was stirred at 60 °C for 2 hours. The volatiles were removed under vacuo, the residue was diluted with ethyl acetate and washed with sat. NaCl. The organic phase was dried with magnesium sulfate, filtered, and concentrated. The crude material was purified by silica gel column chromatography eluting with ethyl acetate and hexanes (1 :2) to give tert-butyl (3R,4R,5S)-4-(tert-butyldimethylsilyloxy)-5-methyl-l-(3-nitropyridin-4-yl)piperidin-3- ylcarbamate in 76% yield. LC/MS = 467.3 (M+H), Rt = 1.09 min.

Synthesis of tert-butyl (3R,4R,5S)-l-(3-aminopyridin-4-yl)-4-(tert- butyldimethylsilyloxy)-5 -methylpiperidin-3 -ylcarbamate

OTBS

BocHN

A solution of tert-butyl (3R,4R,5S)-4-(tert-butyldimethylsilyloxy)-5-methyl-l-(3- nitropyridin-4-yl)piperidin-3-ylcarbamate (1.0 equiv.) in MeOH (0.05 M) was degassed with nitrogen for 20 min. 10% Pd/C (0.2 equiv.) was added to the mixture and the solution was stirred under a hydrogen balloon for 3 hours. The reaction was filtered and the filtrate was concentrated under reduced pressure to give tert-butyl (3R,4R,5S)-l-(3- aminopyridin-4-yl)-4-(tert-butyldimethylsilyloxy)-5 -methylpiperidin-3 -ylcarbamate as the desired product in 94% yield. LC/MS = 437.4 (M+H), Rt = 1.08 min. 1H-NMR (300

MHz, CDCls): δ 8.01 (s, 1H), 7.95 (d, J = 6.0 Hz, 1H), 6.76 (d, J = 6.0 Hz, 1H), 4.44 (b s, 1H), 3.74 (br s, 2H), 3.59-3.55 (m, 1H), 3.25-3.13 (m, 2H), 2.47-2.35 (m, 2H), 1.89 (b s, 2H), 1.44 (s, 9H), 1.04 (d, J = 6.0 , 3H), 0.92 (s, 9H), 0.13 (d, J = 9.0, 6H).

Synthesis of 5-methyl-3-oxocyclohex- 1 -enyltrifluoromethanesulfonate

To a solution of 5-methylcyclohexane-l,3-dione (1.0 equiv.) in DCM (0.5M) was added Na₂C0₃ (1.1 equiv.) and cooled to 0 °C. Added Tf₂0 (1.0 equiv.) in DCM (5.0 M) dropwise over 1 hr at 0°C under a nitrogen atmosphere. Upon addition, the reaction was stirred for 1 hr at room temperature (dark red solution). The solution was filtered and the filtrate was quenched by careful addition of saturated NaHC0₃ with vigorous stirring until pH=7. The solution was transferred to a separatory funnel and the layers were separated. The organic layer was washed with brine, dried with Na₂S0₄, filtered, concentrated under vacuo and dried under high vacuum for 15 min to yield 5-methyl-3- oxocyclohex-l-enyl trifluoromethanesulfonate as light yellow oil in 78% yield. The triflate decomposes upon storage and should be used immediately for the next reaction.

LC/MS=259.1/300.1 (M+H and M+CH₃CN); Rt = 0.86 min, LC = 3.84 min. 1H-NMR (400 MHz, CDC1₃) δ ppm: 6.05 (s, 1H), 2.70 (dd, J=17.2, 4.3, 1H), 2.53 (dd, J=16.6, 3.7, 1H), 2.48-2.31 (m, 2H), 2.16 (dd, J=16.4, 11.7, 1H), 1.16 (d, J=5.9, 3H). Synthesis of 5-methyl-3-(4 A5,5-tetramethyl-l ,3,2-dioxaborolan-2-yl)cyclohex-2-enone

To a solution of 5-methyl-3-oxocyclohex-l-enyl trifluoromethanesulfonate (1.0 equiv.) in degassed dioxane (0.7 M) was added bis(pinacolato)diboron (2.0 equiv.), KOAc (3.0 equiv.), and Pd(dppf)Cl₂-DCM (0.03 equiv.). The reaction was heated to 80 °C for 10 h (initial heating at large scale results in exothermic formation of an orange foam on top of the solution, the heating bath should be removed until the foam retracts, reheating to 80 °C at this point appears to be fine), then cooled to room temperature and filtered through a coarse frit glass funnel. The cake was rinsed with more dioxane and the filtrate solution was used for the next step without further purification. LC/MS = 155.1 (M+H of boronic acid); Rt = 0.41 min, LC = 1.37 min.

Synthesis of 5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enone

To a solution of 5-methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)cyclohex-2-enone (1.0 equiv.) in degassed dioxane (0.5 M) and 2M Na₂C0₃ (2 equiv.) was added 4-chloro-3-nitropyridine (1.3 equiv.) and Pd(dppf)Cl₂-DCM (0.05 equiv.). The reaction was placed under a reflux condenser and heated in an oil bath to 110°C for 1 h. Cooled to room temperature, filtered through a pad of Celite, washed the pad with ethyl acetate and concentrated the filtrate under vacuo. The residue was further pumped at 80 °C on a rotary evaporator for one hour to remove boronate by-products (M+H = 101) via sublimation. The residue was partitioned between brine and ethyl acetate, and the layers were separated, the aqueous phase was further extracted with ethyl acetate (4x), the organics were combined, dried over sodium sulfate, filtered, and concentrated. The crude was purified via silica gel chromatography loading in DCM and eluting with 2-50% ethyl acetate and hexanes. The pure fractions were concentrated in vacuo to yield an orange oil. The oil was placed under high vacuum (-500 mtorr) with seed crystals overnight to yield an orange solid. The solid was further purified via trituration in hexanes to yield 5- methyl-3-(3-nitropyridin-4-yl) cyclohex-2-enone (48% 2 steps). LC/MS = 233.2 (M+H); Rt = 0.69 min, LC = 2.70 min. 1H-NMR (400 MHz, CdCl₃) δ ppm: 9.31 (s, IH), 8.88 (d, J=5.1, IH), 7.30 (d, J=5.1, IH), 6.00 (d, J=2.4, IH), 2.62 (dd, J=16.4, 3.5, IH), 2.53-2.34 (m, 3H), 2.23 (dd, J=16.1, 11.7, IH), 1.16 (d, J=6.3, 3H).

Synthesis of cis-(+/-)-5-methyl-3-(3-nitropyridin-4-yl)cvclohex-2-enol

To a solution of 5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enone (1.0 equiv.) in EtOH (0.3 M) was added CeCl₃-7H₂0 (1.2 equiv.). The reaction was cooled to 0°C, then NaBH₄ (1.2 equiv.) was added in portions. Stirred for 1 h at 0°C, then quenched by adding water, concentrated to remove the EtOH, added EtOAc, extracted the organics, washed with brine, then dried with Na₂S0₄, filtered and concentrated to yield cis-(+/-)-5- methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enol (94%). LC/MS = 235.2 (M+H), LC = 2.62 min.

Synthesis of (+/-)-4-(5-methylcyclohexa- 1 ,3-dienyl)-3-nitropyridine

To a solution of (+/-)-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enol (1.0 equiv.) in dioxane (0.1M) was added p-TSA (1.0 equiv.), and the reaction was stirred at 100 °C for 3 h. The solution was cooled to room temperature, then passed through a pad of neutral alumina eluting with EtOAc to yield (+/-)-4-(5-methylcyclohexa-l,3-dienyl)-3- nitropyridine as a yellow oil in 68% yield. LC/MS = 217.1 (M+H), LC = 3.908 min.

Synthesis of (+/-)-6-bromo-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enol

To a solution of 4-(5-methylcyclohexa-l,3-dienyl)-3-nitropyridine (1.0 equiv.) in THF and water (1 : 1, 0.13 M) was added NBS (1.5 equiv.) and the reaction was stirred at room temperature for 30 min. Upon completion, ethyl acetate and water were added to the reaction, the organic phase was dried with brine, then sodium sulfate, filtered, and concentrated. The crude material was purified via silica gel column chromatography eluting with ethyl acetate and hexanes (1 : 1) to give (+/-)-6-bromo-5-methyl-3-(3- nitropyridin-4-yl)cyclohex-2-enol as a yellow oil in 80% yield. LC/MS = 315.0/313.0 (M+H), LC = 2.966 min.

Synthesis of (+/-)-2-azido-6-methyl-4-(3-nitropyridin-4-yl)cyclohex-3-enol

To a solution of (+/-)-6-bromo-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enol (1.0 equiv.) in THF (0.1 M) was added potassium tert-butoxide (1.5 equiv.). The reaction turned from orange to black almost immediately. By TLC, the formation of product is clean in 30 min. Quenched by adding saturated ammonium chloride and ethyl acetate. The organic phase was dried with brine, then sodium sulfate, filtered, and concentrated. The crude product was dissolved in ethanol and water (3: 1, 0.1 M), and ammonium chloride (2.0 equiv) and sodium azide (2.0 equiv.) were added. The dark orange reaction was stirred at room temperature overnight. The conversion to product is clean as indicated by LC/MS. The reaction was concentrated to remove the ethanol, ethyl acetate and water were added, and the organic phase was dried with sodium sulfate, filtered, and concentrated. The crude material was purified via silica gel column chromatography eluting with ethyl acetate and hexanes (1 : 1) to give (+/-)-2-azido-6-methyl-4-(3- nitropyridin-4-yl)cyclohex-3-enol in 55% yield. LC/MS = 276.0 (M+H), LC = 2.803 min.

Synthesis of (+/-)-tert-butyl 6-hvdroxy-5-methyl- 3-(3-nitropyridin-4-yl)cyclohex-2-enylcarbamate

To a solution of (+/-)-2-azido-6-methyl-4-(3-nitropyridin-4- yl)cyclohex-3-enol (1.0 equiv.) in pyridine and ammonium hydroxide (8:1, 0.08 M) was added trimethylphosphine (3.0 equiv.) and the brown solution was stirred at room temperature for 2 h. Upon completion, EtOH was added and the solution was concentrated in vacuo. More ethanol was added and the reaction was concentrated again. Dioxane and sat. NaHC0₃ (1 : 1, 0.08 M) were added to the crude, followed by Boc₂0 (1.0 equiv.). Stirred the reaction mixture at room temperature for 2h, then added water and ethyl acetate. The organic phase was dried with MgSC^, and concentrated. The crude product was purified via silica gel column chromatography eluting with ethyl acetate and hexanes (1 : 1) to afford (+/-)-tert-butyl 6-hydroxy-5-methyl-3-(3-nitropyridin- 4-yl)cyclohex-2-enylcarbamate (59%). LC/MS = 350.1 (M+H), Rt: 0.76 min.

Synthesis of (+/-)-2-(tert-butoxycarbonylamino)-6-methyl- 4-(3-nitropyridin-4-yl)cyclohex-3-enyl acetate

To a solution of (+/-)-tert-butyl 6-hydroxy-5-methyl-3-(3-nitropyridin- 4-yl)cyclohex-2-enylcarbamate (1.0 equiv.) in pyridine (0.1 M) was added Ac₂0 (2.0 equiv.) and the reaction was stirred at room temperature overnight. Upon completion, the reaction was concentrated to dryness, then worked-up with ethyl acetate and water. The organic phase was dried with brine, then sodium sulfate, filtered, and concentrated to give (+/ -)-2-(tert-butoxycarbonylamino)-6-methyl-4-(3 -nitropyridin-4-yl)cyclohex-3 -enyl acetate in 94% yield. LC/MS = 392.2 (M+H), Rt = 0.94 min.

Synthesis of (+/-)-4-(3-aminopyridin-4-yl)-2-(tert-butoxycarbonylamino)-6- methylcyclohexyl acetate

To a degassed solution of (+/-)-2-(tert-butoxycarbonylamino)-6-methyl- 4-(3-nitropyridin-4-yl)cyclohex-3-enyl acetate (1.0 equiv.) in MeOH and EtOAc (1 : 1, 0.1 M) was added 10%> Pd/C (0.1 equiv.) and the reaction was stirred at room temperature under a hydrogen balloon for 3 days. Upon completion, the solution was filtered through a pad of Celite, the pad was washed with ethyl acetate and the filtrate was concentrated. The crude material contained about 10% of the undesired isomer. The crude was dissolved in ethyl acetate (-20%) and hexanes and heated until all dissolved. The solution was allowed to sit at room temperature for 2 days. The precipitate was then collected to give (+/-)-4-(3-aminopyridin-4-yl)-2-(tert-butoxycarbonylamino)-6- methylcyclohexyl acetate as the pure product in 59% yield. LC/MS = 364.3 (M+H), Rt = 0.63 min. Synthesis of 4-(3-(tert-butyldimethylsilyloxy)-5-methylcvclohex- 1 -enyl)-3-nitropyridine

To a solution of 5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enol (1.0 equiv.) in DMF (0.5 M) was added imidazole (4.0 equiv.) and TBDMSC1 (2.5 equiv.). After stirring for 18 hours the solution was portioned between EtOAc and H₂0 and separated. After washing further with H₂0 (3x) and NaCl (sat.), drying over MgSC^, filtering and removal of solvents, 4-(3-(tert-butyldimethylsilyloxy)-5-methylcyclohex-l- enyl)-3-nitropyridine was obtained (85%). LC/MS = 349.2 (M+H), LC = 5.99 min.

Synthesis of 4-(3-(tert-butyldimethylsilyloxy)-5-methylcyclohex- 1 -enyl)pyridin-3 -amine

A heterogeneous solution of 4-(3-(tert-butyldimethylsilyloxy)-5- methylcyclohex-l-enyl)-3-nitropyridine (1.0 eq.) and iron (6.0 eq) in acetic acid, at a concentration of 0.4 M, was stirred vigorously for 2 hours. The mixture was then passed through a celite pad, eluting with MeOH. Upon removal of the volatiles in vacuo, the residue was dissolved in EtOAc, washed with Na₂C0_{3 (sa )}, NaCl_{(sa )}, was dried over MgS0₄, was filtered and the volatiles were removed in vacuo yielding 4-(3-(tert- butyldimethylsilyloxy)-5-methylcyclohex-l-enyl)pyridin-3 -amine (78%). LCMS (m/z): 319.3 (MH⁺); LC R, = 3.77 min. Synthesis of 4-(3 -(tert-butyldimethylsilyloxy)-5 -methylcvclohexyl)pyridin-3 -amine

To a solution of 4-(3-(tert-butyldimethylsilyloxy)-5-methylcyclohex-l- enyl)-3-nitropyridine (1.0 equiv.) in methanol, at a concentration of 0.1 M, was added 10% palladium on carbon (0.1 eq.). The resultant heterogeneous solution was put under an atmosphere of hydrogen and was stirred for 15 hours. At this time the mixture was filtered through a pad of celite eluting with methanol. The volatiles were removed in vacuo yielding 4-(3 -(tert-butyldimethylsilyloxy)-5 -methylcyclohexyl)pyridin-3 -amine (90%). LCMS (m/z): 321.3 (MH⁺); LC R, = 3.85 min.

Synthesis of cis (+/-) benzyl 4-3-(tert-butyldimethylsilyloxy)-5- methylcvclohexyDpyridin-3-ylcarbamate

To a solution of cis-(+/-)-4-(3-(tert-butyldimethylsilyloxy)-5- methylcyclohexyl)pyridin-3 -amine in dichloromethane at a concentration of 0.5 M was added benzyl 2,5-dioxopyrrolidin-l-yl carbonate (1.1 equiv.) and DMAP (0.05 equiv.). After stirring for 16 hours at rt, additional benzyl 2,5-dioxopyrrolidin-l-yl carbonate (0.55 equiv.) and DMAP (0.03 equiv.) were added. After stirring for an additional 24 hours at rt, additional benzyl 2,5-dioxopyrrolidin-l-yl carbonate (0.1 equiv.) and DMAP (0.03 equiv.) were added. After stirring for 18 more hours the solution was partitioned between EtOAc and Na₂C03(_sat.) and separated. Upon further washing with Na₂C03(_sat.) (2x) and NaCl(_sat.), drying over MgS0₄, filtering and removal of solvents, cis (+/-) benzyl 4-3 -(tert-butyldimethylsilyloxy)-5 -methylcyclohexyl)pyridin-3 -ylcarbamate was obtained. The crude material was used as is. LC/MS = 455.3 (M+H), LC = 4.39 min.

Synthesis of cis-(+/-)benzyl 4-(3-hydroxy-

5 -methylcvclohexyl)pyridin-3 -ylcarbamate

A solution of cis (+/-) benzyl 4-3-(tert-butyldimethylsilyloxy)-5- methylcyclohexyl)pyridin-3-ylcarbamate in 1 :2: 1 6N HCl/THF/MeOH at a concentration of 0.1 M was stirred at rt for 6 hours. The pH was than adjusted to pH=7 by addition of 6N NaOH and the volatiles were removed in vacuo. The aqueous layer was extracted with EtOAc and the organic was washed with NaCl_(sat), dried over MgS0₄, filtered and upon removal of the volatiles in vacuo, cis-(+/-)benzyl 4-(3-hydroxy-5- methylcyclohexyl)pyridin-3 -ylcarbamate was obtained. The crude material was used as is. LC/MS = 341.2 (M+H), LC = 2.38 min. Synthesis of cis (+/-)-benzyl 4-(3-methyl-5-oxocvclohexyl)pyridin-3 -ylcarbamate

To a 0 °C solution of cis-(+/-)-benzyl 4-(3-hydroxy-5-methyl- cyclohexyl)pyridin-3 -ylcarbamate in wet CH₂CI₂ at a concentration of 0.16 M was added Dess-Martin Periodinane (1.5 equiv.) and the solution was stirred for 18 hours as it warmed to rt. The solution was partitioned between EtOAc and 1 : 1 10% Na₂S203/NaHC03(sat.) and separated. Upon further washing with 1 : 1 10% Na₂S₂0₃/NaHC0_3(sat.₎ (2x) and NaCl_(sat), drying over MgS0₄, filtering, removal of solvents and purification by silica gel chromatography (75-100% EtOAc/hexanes), cis- (+/-)-benzyl-4-(3-methyl-5-oxocyclohexyl)pyridin-3-ylcarbamate was obtained as a white solid (53%, 5 steps). LC/MS = 339.2 (M+H). Synthesis of cis-(+/-)- benzyl 4-(-3-(benzylamino)-

5 -methylcyclohexy l)pyridin-3 -ylcarbamate

A solution of cis-(+/-)-benzyl-4-(3-methyl-5-oxocyclohexyl)pyridin-3- ylcarbamate (1.0 equiv) and benzylamine (3.0 equiv) in MeOH, at a concentration of 0.25 M, was stirred at rt for 2 hours. Upon cooling in a -78 °C bath, LiBH₄ (1.1 equiv, 2.0 M in THF) was added and the solution was allowed to warm to rt with stirring over 16 hours. The solution was partitioned between EtOAc and NaHC0_{3(sat ),} separated, washed further with NaHC0_3(sat.₎ and NaCl_(sat), dried over MgS0₄, filtered and after removal of volatiles in vacuo, cis-(+/-)- benzyl 4-(-3-(benzylamino)-5-methylcyclohexyl)pyridin-3- ylcarbamate was obtained as a 4: 1 mixture of isomers, with the all cis as predominant LC/MS = 430.3 (M+H), LC = 0.62 min.

Synthesis of cis (+/-)-tert-butyl (-3-(3-aminopyridin-4-yl)-5-methylcvclohexylcarbamate

To a solution of cis-(+/-)- benzyl 4-(-3-(benzylamino)-5- methylcyclohexyl)pyridin-3-ylcarbamate was (1.0 equiv.) in methanol, at a concentration of 0.07 M, was added 20% palladium hydroxide on carbon (0.2 eq.). The resultant heterogeneous solution was put under an atmosphere of hydrogen and was stirred for 14 hours. At this time the reaction was purged with Ar, Boc₂0 (1.0 equiv.) was added and the solution was stirred for 8 hours. Additional Boc₂0 (1.0 equiv.) was added and the solution was stirred for 16 more hours. At this time the mixture was filtered through a pad of celite eluting with methanol. Upon removal of volatiles in vacuo, purification by silica gel chromatography (2.5-2.5 MeOH/CH₂Cl₂ with 0.1% DIEA) and recrystallization from 10% EtOAc/hexanes yielded cis (+/-)-tert-butyl (-3-(3-aminopyridin-4-yl)-5- methylcyclohexylcarbamate (49%). LCMS (m/z): 306.3 (MH⁺), LC R, = 2.59 min . Pure enantiomers could be obtained by chiral chromatography.

Synthesis of ethyl 2-amino-2-cyanoacetate

To a solution of ethyl 2-cyano-2-(hydroxyimino)acetate (leq) in 70 mL of water and 56 mL of aq. sat. sodium bicarbonate was added portionwise throughout 10 minutes Na₂S₂0₄ (2.8 eq) The reaction mixture was stirred at room temperature for 1 hour. The solution was saturated with sodium chloride, extracted with methylene chloride (300mL x 3) and then the combined organic layers were dried over anhydrous Na₂S0₄, filtered, and concentrated in vacuo to give ethyl 2-amino-2-cyanoacetate, which was used to next step without further (55%). LC/MS (m/z): 129.0 (MH⁺), R_t: 0.25 min.

Synthesis of ethyl 2-cyano-2-formamidoacetate

A mixture of acetic anhydride (1.80 eq.) and formic acid (1.85 eq.) was heated to 55 °C for 3 h. The reaction mixture was then cooled to 0°C and a solution of ethyl 2-amino-2- cyanoacetate (1.00 eq.) in THF (0.2 M) was added dropwise. The mixture was allowed to warm to RT for 18 h and then concentrated in vacuo. The crude residue was purified by flash column chromatography eluting with (EtOAc : hexanes= 1 : 4) to give ethyl 2- cyano-2-formamidoacetate in 70% yield. 1H NMR (400 MHz, <cdcl3>) δ ppm 1.38 (t, J=7.24 Hz, 3 H) 4.39 (q, J=7.04 Hz, 2 H) 5.48 - 5.64 (m, 1 H) 6.38 - 6.67 (m, 1 H) 7.26 (s, 1 H) 8.32 (s, 1 H).

Synthesis of ethyl 5-aminothiazole-4-carboxylate

To a solution of 2-cyano-2-formamidoacetate (1.00 eq.) in pyridine (0.1 M) was added Lawesson's reagent (1.20 eq.) at RT. The resulting mixture was heated to 120 °C for 16 h. The reaction mixture was then concentrated in vacuo to yield a black oil. The oil was further purified by flash column chromatography eluting with (EtOAc : heptanes= 1 : 1) to afford ethyl 5-aminothiazole-4-carboxylate in 44% yield. LC/MS (m/z): 173.0 (MH⁺), R,: 0.40 min. 1H NMR (400 MHz, <cdcl3>) δ ppm 1.35 - 1.46 (m, 3 H) 1.52 - 1.72 (m, 1 H) 4.34 - 4.45 (m, 2 H) 5.83 - 6.18 (m, 2 H) 7.22 - 7.30 (m, 1 H) 7.88 (s, l H)

Synthesis of ethyl 5-((tert-butoxycarbonyl)amino)thiazole-4-carboxylate

To a solution of ethyl 5-aminothiazole-4-carboxylate (1.00 eq.) in CH₃CN (0.1 M) at RT was added Boc-anhydride (1.1 eq.) at RT. The mixture was stirred at RT for 68 h. The reaction mixture was then concentrated in vacuo and purified by flash column chromatography eluting with (EtOAc : heptanes= 1 : 5) to afford ethyl 5-((tert- butoxycarbonyl)amino)thiazole-4-carboxylate in 75% yield. LC/MS (m/z): 273.1 (MH⁺), Rt = 0.90 min). 1H NMR (400 MHz, <cdcl3>) δ ppm 1.45 (t, J=7.04 Hz, 3 H) 1.50 - 1.61 (m, 10 H) 4.45 (q, J=7.17 Hz, 2 H) 8.28 (s, 1 H) 9.92 (br. s., 1 H)

Synthesis of ethyl 2-bromo-5-((tert-butoxycarbonyl)amino)thiazole-4-carboxylate

To a solution of ethyl 5-(tert-butoxycarbonylamino)thiazole-4-carboxylate (1.0 equiv.) in DCM (0.20 M) was added NBS (1.6 equiv) at RT. The resulting mixture was stirred at RT for 2 hrs. The reaction mixture was then concentrated in vacuo to give ethyl 2-bromo-5-((tert-butoxycarbonyl)amino)thiazole-4-carboxylate in 100% yield and utilized in the next reaction without further purification. LC/MS = 352.9 (MH+), Rt = 1.12 min. 1H NMR (400 MHz, <cdcl3>) δ ppm 1.33 - 1.46 (m, 3 H) 1.48 - 1.61 (m, 9 H) 4.34 - 4.52 (m, 2 H) 9.92 (br. s., 1 H)

Method 1

Synthesis of ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4- methoxyphenyl)thiazole-4-carboxylate

To a solution of ethyl 2-bromo-5-((tert-butoxycarbonyl)amino)thiazole-4-carboxylate (1.0 equiv.) in THF and water (10: 1, 0.1 M) was added 2,6-difluoro-4-methoxyphenyl)boronic acid (2.0 equiv.) and potassium fluoride (3.00 equiv.). The reaction was degassed with nitrogen, then Pd₂(dba)₃ (0.20 equiv.) and tri-tert-butylphosphine (0.4 equiv.) were added and the reaction was heated to 100 °C under microwave irradiation for 20 min. LC/MS analysis indicated complete conversion of the starting material to product. The reaction was cooled to room temperature then quenched with water and diluted with EtOAc. The aqueous layer was separated then extracted with EtOAc. The combined organics were dried over MgSC^ and concentrated in vacuo to yield a brown oil. The crude product was purified by ISCO flash chromatography eluting with ethyl acetate and hexanes (0% to 30% ethyl acetate) to provide ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4- methoxyphenyl)thiazole-4-carboxylate as the desired product as a white solid in 73% yield. LC/MS = 415.0 (M+H), Rt = 1.26 min; 1H NMR (400 MHz, <cdcl3>) δ ppm 1.45 (t, J=7.24 Hz, 3 H) 1.48 - 1.63 (m, 11 H) 3.74 - 3.89 (m, 3 H) 4.40 - 4.56 (m, 2 H) 6.43 - 6.65 (m, 2 H) 9.95 - 10.07 (m, 1 H).

Method 2

Synthesis of 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-methoxyphenyl)thiazole-4- carboxylic acid

To a solution of methyl ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4- methoxyphenyl)thiazole-4-carboxylate (1.0 equiv.) in THF/MeOH (2: 1, 0.3 M) was added LiOH (10.0 equiv.) and the reaction was stirred at 55 °C for 36 hour. The solution was quenched with 2N HC1, extracted with ethyl acetate, washed with brine, dried with sodium sulfate, filtered and concentrated to give 5-((tert-butoxycarbonyl)amino)-2-(2,6- difluoro-4-methoxyphenyl)thiazole-4-carboxylic acid in 99% yield. LC/MS = 386.9 (M+H-^lBu), Rt = 1.00 min. 1H NMR (400 MHz, <dmso>) d ppm 1.48 (s, 9 H) 3.29 (br. s., 9 H) 3.76 - 3.90 (m, 3 H) 6.84 - 6.98 (m, 2 H) 10.06 (br. s., 1 H)

Method 3 Synthesis of 2-(2,6-difluoro-4-methylphenyl)-4,4,5,5-tetramethyl-l ,3,2- dioxaboroane

To a solution of l,3-difluoro-5-methylbenzene (1.0 eq) in dry THF (0.2M) under an atmosphere of N₂ at -78°C was added n-butyllithium (leq, 1.6M in hexanes) slowly keeping the internal temperature below -65°C. The reaction was stirred for 2 hrs at - 78°C, followed by the addition of 2-isopropoxy-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (1.15eq). The reaction was allowed to warm to room temperature. Upon completion, the reaction was quenched with NaHC0_{3 (sat}.₎ and extracted with EtOAc. The organics were washed with brine, dried over Na₂S0₄, filtered and concentrated to yield a 2-(2,6- difluoro-4-methylphenyl)-4,4,5,5-tetramethyl-l,3,2-dioxaboroane as a white solid in 92%. 1H NMR (400 MHz, <cdcl3>) δ ppm 6.67 (dd, J=9.39, 0.78 Hz, 2 H), 2.34 (s, 3 H), 1.38 (s, 12 H).

Synthesis of ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4- methylphenyl)thiazole-4-carboxylate

Method 1 was followed using ethyl 2-bromo-5-((tert-butoxycarbonyl)amino)thiazole-4- carboxylate (1.0 equiv.) and 2-(2,6-difluoro-4-methylphenyl)-4,4,5,5-tetramethyl-l,3,2- dioxaboroane (2.00 equiv.) to give ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro- 4-methylphenyl)thiazole-4-carboxylate as a solid in 70% yield. LC/MS =399.2 (M+H), Rt = 1.19 min.

Synthesis of 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4- methylphenyl)thiazole-4-carboxylic acid

To a solution of ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4- methylphenyl)thiazole-4-carboxylate (l .Oeq) in THF/EtOH (2: 1, 0.3 M) was added LiOH (10.0 equiv.) and the reaction was stirred at 55 °C for 28 hour. The solution was quenched with 2N HCl, extracted with ethyl acetate, washed with brine, dried with sodium sulfate, filtered and concentrated to give 5-((tert-butoxycarbonyl)amino)-2-(2,6- difluoro-4-methylphenyl)thiazole-4-carboxylic acid in 64% yield. LC/MS = 371.2 (M+H), Rt = 0.98 min.

Synthesis of l,3-difluoro-5-isopropoxybenzene

To a solution of 3,5-difluorophenol (1.0 equiv.) in DMF (0.26 M) was added potassium carbonate (2.2 equiv.) followed by 2-iodopropane (1.1 equiv.) and the reaction was stirred overnight at room temperature. The reaction was poured into a separatory funnel and diluted with a 3: 1 (v/v) solution of EtOAc:heptanes. The organic phase was washed with water, then sat'd NaHC03. The remaining organic phase was dried over MgS04, filtered and concentrated in vacuo to provide l,3-difluoro-5-isopropoxybenzene in 88% yield. 1H NMR (400 MHz, <cdcl3>) δ ppm 1.33 (d, J=6.26 Hz, 6 H), 4.48 (dt, J=11.93, 6.16 Hz, 1 H), 6.31 - 6.47 (m, 3 H).

Synthesis of 2-(2,6-difluoro-4-isopropoxyphenyl)-4,4,5,5-tetramethyl-l ,3,2- dioxaborolane

Method 3 was followed using 2-isopropoxy-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (2.2 equiv.), butyllithium (1.2 equiv.) and l,3-difluoro-5-isopropoxybenzene (1.0 equiv.) to give 2-(2,6-difluoro-4-isopropoxyphenyl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane in 99% yield. 1H NMR (400 MHz, <cdcl3>) δ ppm 1.24 (s, 12 H), 1.31 - 1.33 (m, 6 H), 4.43 - 4.56 (m, 1 H), 6.31 - 6.44 (m, 2 H). Synthesis of ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4- isopropoxy henyl)thiazole-4-carboxylate

Method 1 was followed using methyl ethyl 2-bromo-5-((tert- butoxycarbonyl)amino)thiazole-4-carboxylate (1.0 equiv.) and 2-(2,6-difluoro-4- isopropoxyphenyl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (2.0 equiv.) at 90 °C under microwave irradiation for 15 min to give ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6- difluoro-4-isopropoxyphenyl)thiazole-4-carboxylate in 62% yield. LC/MS =443.2 (MH+), Rt = 1.29 min.

Synthesis of 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4- isopropoxyphenyl)thiazole-4-carboxylic acid

Method 2 was followed using ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4- isopropoxyphenyl)thiazole-4-carboxylate to give 5-((tert-butoxycarbonyl)amino)-2-(2,6- difluoro-4-isopropoxyphenyl)thiazole-4-carboxylic acid in 76% yield. LC/MS = 415.2 (MH+), Rt = 1.11 min.

Synthesis of l-(ethoxymethyl)-3,5-difluorobenzene

To a solution of (3,5-difluorophenyl)methanol (1.0 equiv.) in DMF (0.225 M) at 0°C was added sodium hydride (1.5 eq). the reaction mixture was stirred for 15 min at 0°C before the dropwise addition of ethyl iodide ( 3.0 equiv.). The ice bath was removed and stirring was continued for 18 hours. The reaction mixture was quenched by the addition of water and diluted with 3: 1 EtOAc: heptanes. The combined organics were washed with water, dried over MgSC^, filtered, concentrated and purified by Si02 chromatography to yield l-(ethoxymethyl)-3,5-difluorobenzene in 99%.

Synthesis of 2-(4-(ethoxymethyl)-2,6-difluorophenyl)-4,4,5,5-tetramethyl-l ,3,2- dioxaborolane

To a solution of l-(ethoxymethyl)-3,5-difluorobenzene (l .Oeq) in dry THF (0.2 M) under an atmosphere of N₂ at -78°C was added n-butyllithium (leq, 1.6M in hexanes) slowly keeping the internal temperature below -65°C. The reaction was stirred for 1 hr at -78°C, followed by the addition of 2-(4-(ethoxymethyl)-2,6-difluorophenyl)-4,4,5,5-tetramethyl- 1,3,2-dioxaborolane (2.1 eq). The reaction was allowed to warm to room temperature. Upon completion, the reaction was quenched with NaHC0_{3 (sat}.₎ and extracted with EtOAc. The organics were washed with brine, dried over Na₂S0₄, filtered and concentrated to yield 2-(4-(ethoxymethyl)-2,6-difluorophenyl)-4,4,5,5-tetramethyl- 1,3,2-dioxaborolane in 71%.

Synthesis of ethyl 5-((tert-butoxycarbonyl)amino)-2-(4-(ethoxymethyl)-2,6- difluorophenyl)thiazole-4-carboxylate

Method 1 was followed using ethyl 2-bromo-5-((tert-butoxycarbonyl)amino)thiazole-4- carboxylate (1.0 equiv.) and 2-(4-(ethoxymethyl)-2,6-difluorophenyl)-4,4,5,5- tetramethyl-l,3,2-dioxaborolane (2.0 equiv.) at 100 °C under microwave irradiation for 20 min to give ethyl 5-((tert-butoxycarbonyl)amino)-2-(4-(ethoxymethyl)-2,6- difhiorophenyl)thiazole-4-carboxylate in 73% yield.. LC/MS =443.2 (MH+), Rt = 1.29 mm.

Synthesis of 5-((tert-butoxycarbonyl)amino)-2-(4-(ethoxymethyl)-2,6- difluorophenyl)thiazole-4-carboxylic acid

Method 2 was followed using ethyl 5-((tert-butoxycarbonyl)amino)-2-(4-(ethoxymethyl)- 2,6-difluorophenyl)thiazole-4-carboxylate (1.0 eq) to give 5-((tert- butoxycarbonyl)amino)-2-(4-(ethoxymethyl)-2,6-difluorophenyl)thiazole-4-carboxylic acid in 76% yield. LC/MS = 415.2 (MH+), Rt = 1.06 min.

Synthesis of 4-(3,5-difluorophenoxy)tetrahydro-2H-pyran

To a solution of 3,5-difluorophenol (1.0 equiv.), tetrahydro-2H-pyran-4-ol (1.2 equiv.), and triphenylphosphine (2.0 equiv.) in THF (0.33 M) at 0 °C was added DIAD (2.0 equiv.) dropwise. The reaction mixture was stirred at rt overnight. The mixture was concentrated and purified by flash chromatography over silica gel (heptanes: ethyl acetate gradient) to give 4-(3,5-difluorophenoxy)tetrahydro-2H-pyran in 90 % yield. 1H NMR (400 MHz, <cdcl3>) δ ppm 1.72 - 1.84 (m, 2 H), 1.96 - 2.09 (m, 2 H), 3.59 (ddd, J=1 1.64, 8.31 , 3.52 Hz, 2 H), 3.90 - 4.04 (m, 2 H), 4.44 (tt, J=7.78, 3.77 Hz, 1 H), 6.32 - 6.53 (m, 3 H). Synthesis of 2-(2,6-difluoro-4-((tetrahvdro-2H-pyran-4-yl)oxy)phenyl)-4,4,5,5- tetramethyl- 1 ,3 ,2-dioxaborolane

Method 3 was followed using 2-isopropoxy-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (1.5 equiv.), butyllithium (1.3 equiv.) and 4-(3,5-difluorophenoxy)tetrahydro-2H-pyran (1.0 equiv.) to give 2-(2,6-difluoro-4-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)-4,4,5,5- tetramethyl-l,3,2-dioxaborolane in 33% yield. 1H NMR (400 MHz, <cdcl3>) δ ppm 1.21 - 1.34 (m, 12 H), 1.78 (dtd, J=12.72, 8.31, 8.31, 3.91 Hz, 2 H), 1.93 - 2.09 (m, 2 H), 3.59 (ddd, J=11.64, 8.31, 3.13 Hz, 2 H), 3.89 - 4.01 (m, 2 H), 4.48 (tt, J=7.78, 3.77 Hz, 1 H), 6.40 (d, J=9.39 Hz, 2 H).

Synthesis of ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-((tetrahvdro-2H- pyran-4-yl)oxy)phenyl)thiazole-4-carboxylate

Method 1 was followed using methyl ethyl 2-bromo-5-((tert- butoxycarbonyl)amino)thiazole-4-carboxylate (0.8 equiv.) and 2-(2,6-difluoro-4- ((tetrahydro-2H-pyran-4-yl)oxy)phenyl)-4,4,5,5-tetramethyl-l ,3,2-dioxaborolane (1.0 equiv.) at 100 °C under microwave irradiation for 20 min to give ethyl 5-((tert- butoxycarbonyl)amino)-2-(2,6-difluoro-4-((tetrahydro-2H-pyran-4- yl)oxy)phenyl)thiazole-4-carboxylate in 67% yield. LC/MS =485.0 (MH+), Rt = 1.23 min. 1H NMR (400 MHz, <cdcl3>) δ ppm 0.88 (t, J=6.85 Hz, 1 H) 1.24 (s, 9 H) 1.26 - 1.29 (m, 4 H) 1.73 - 1.87 (m, 2 H) 1.94 - 2.09 (m, 3 H) 3.52 - 3.66 (m, 2 H) 3.91 - 4.03 (m, 2 H) 4.41 - 4.54 (m, 2 H) 6.45 - 6.62 (m, 2 H) 10.02 (s, 1 H)

Synthesis of 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-((tetrahvdro-2H-pyran-4- yl)oxy)phenyl)thiazole-4-carboxylic acid

Method 2 was followed using ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4- ((tetrahydro-2H-pyran-4-yl)oxy)phenyl)thiazole-4-carboxylate to give 5-((tert- butoxycarbonyl)amino)-2-(2,6-difluoro-4-((tetrahydro-2H-pyran-4- yl)oxy)phenyl)thiazole-4-carboxylic acid in 70% yield. LC/MS = 457.0 (MH+), Rt = 1.03 min. 1H NMR (400 MHz, <dmso>) δ ppm 1.48 (s, 8 H) 1.51 - 1.64 (m, 2 H) 1.91 - 2.04 (m, 2 H) 3.37 - 3.60 (m, 5 H) 3.82 (d, J=11.74 Hz, 2 H) 4.63 -4.77 (m, 1 H) 6.98 (d, J=10.96 Hz, 2 H) 9.95 - 10.13 (m, 1 H)

Synthesis of l,3-difluoro-5-(2-methoxyethoxy)benzene

To a solution of 3,5-difluorophenol (1.0 equiv.), 2-methoxyethanol (3.0 equiv.) and triphenylphosphine (3.0 equiv) in THF (0.1 M) was added DIAD (3.0 equiv.). After stirring at rt for 18 hours, the volatiles were removed in vacuo and the residue was purified by Si0₂ chromatography to yield l,3-difluoro-5-(2-methoxyethoxy)benzene in 95%. ^!H NMR (400 MHz, <cdcl3>) δ ppm 6.41-6.47 m (3 H), 4.08 (t, 2H), 3.74 (t, 2H), 3.45 (s, 3 H).

Synthesis of 2-(2,6-difluoro-4-(2-methoxyethoxy)phenyl)-4,4,5,5-tetramethyl- 1,3.2- dioxaborolane

To a solution of l,3-difluoro-5-(2-methoxyethoxy)benzene (l .Oeq) in dry THF (0.2 M) under an atmosphere of N₂ at -78°C was added n-butyllithium (leq, 1.6 M in hexanes) slowly keeping the internal temperature below -65°C. The reaction was stirred for 1 hr at -78°C, followed by the addition of 2-isopropoxy-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (2.1 eq). The reaction was allowed to warm to room temperature. Upon completion, the reaction was quenched with NaHC0_{3 (sat}.₎ and extracted with EtOAc. The organics were washed with brine, dried over Na₂S0₄, filtered and concentrated to yield 2-(2,6-difluoro-

4-(2-methoxyethoxy)phenyl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane. 1H NMR (400 MHz, <cdcl3>) δ ppm 6.42 (d, 2 H), 4.10 (m, 2H), 3.74 (m, 2H), 3.44 (s, 3 H), 1.37 (s, 12 H). Synthesis of ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(2- methoxyethoxy)phenyl)thiazole-4-carboxylate

Method 1 was followed using ethyl 2-bromo-5-((tert-butoxycarbonyl)amino)thiazole-4- carboxylate (1.0 equiv.) and 2-(2,6-difluoro-4-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)- 4,4,5,5-tetramethyl-l,3,2-dioxaborolane (2.0 equiv.) at 90 °C under microwave irradiation for 15 min to give ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(2- methoxyethoxy)phenyl)thiazole-4-carboxylatein 56% yield. LC/MS =459.2 (MH+), Rt = 1.17 min.

Synthesis of 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(2- methoxyethoxy)phenyl)thiazole-4-carboxylic acid

Method 2 was followed using ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4- ((tetrahydro-2H-pyran-4-yl)oxy)phenyl)thiazole-4-carboxylate to give 5-((tert- butoxycarbonyl)amino)-2-(2,6-difluoro-4-(2-methoxyethoxy)phenyl)thiazole-4- carboxylic acid in 88% yield. LC/MS = 431.2 (MH+), Rt = 0.99 min. Synthesis of 3-(3,5-difluorophenyl)oxetan-3-ol

To a solution of l-bromo-3,5-difluorobenzene in THF (0.27 M) under Ar was added Mg turnings (1.6 M). A reflux condenser was attached and the solution was submerged in a 90 °C oil bath and refluxed for two hours. The oxetan-3-one (1.0 equiv.) was added in THF via syringe. The solution was left stirring at rt under Ar overnight. The reaction solution was quenched by addition of NH₄Cl(_sat) and the solution was extracted with EtOAc, washed with NaCl_(sat.₎, dried over MgS0₄, filtered, concentrated and purified by ISCO Si0₂ chromatography (0-100% EtOAc/n-heptanes gradient) to yield 3-(3,5- difhiorophenyl)oxetan-3-ol in 56% yield. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 4.82 (d, J=7.63 Hz, 2 H), 4.91 (d, J=7.63 Hz, 2 H), 7.16 - 7.23 (m, 2 H).

Synthesis of 3-(3,5-difluoro-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- vDphenvDoxetan-3 -ol

Method 3 was followed using 2-isopropoxy-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (2.5 equiv.), butyllithium (2.4 equiv.) and 3-(3,5-difluorophenyl)oxetan-3-ol (1.0 equiv.) to give 3-(3,5-difluoro-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)phenyl)oxetan-3-ol in 79% yield. 1H NMR (400 MHz, <cdcl3>) δ ppml .34 - 1.42 (m, 12 H), 4.79 (d, J=7.63 Hz, 2 H), 4.90 (d, J=7.34 Hz, 2 H), 7.17 (d, J=8.22 Hz, 2 H).

Synthesis of ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(3-hvdroxyoxetan- 3-yl)phenyl)thiazole-4-carboxylate

Method 1 was followed using ethyl 2-bromo-5-((tert-butoxycarbonyl)amino)thiazole-4- carboxylate (1.0 equiv.) and 3-(3,5-difluoro-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)phenyl)oxetan-3-ol (2.0 equiv.) at 100 °C under microwave irradiation for 20 min to give ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(3-hydroxyoxetan-3- yl)phenyl)thiazole-4-carboxylate in 50% yield. LC/MS =457.2 (MH+), Rt = 1.04 min. 1H NMR (400 MHz, <cdcl3>) δ ppm 1.42 - 1.52 (m, 4 H) 1.53 - 1.64 (m, 13 H) 3.25 (br. s., 1 H) 4.43 - 4.55 (m, 2 H) 4.79 (d, J=7.04 Hz, 2 H) 4.95 (d, J=7.04 Hz, 2 H) 7.33 (d, J=9.00 Hz, 2 H) 10.04 (s, 1 H). Synthesis of 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(3-hvdroxyoxetan-3- yl)phenyl)thiazole-4-carboxylic acid

Method 2 was followed using ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(3- hydroxyoxetan-3-yl)phenyl)thiazole-4-carboxylate to give 5-((tert- butoxycarbonyl)amino)-2-(2,6-difluoro-4-(3-hydroxyoxetan-3-yl)phenyl)thiazole-4- carboxylic acid in 76% yield. LC/MS = 429.2 (MH+), Rt = 0.85 min.

Synthesis of ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(3-fluorooxetan-3- yl)phenyl)thiazole-4-carboxylate

To a solution of ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(3- hydroxyoxetan-3-yl)phenyl)thiazole-4-carboxylate (1.0 equiv.) in CH₂CI₂ (0.04 M) at - 78 °C under Ar was added methylDAST (1.0 equiv.). After addition, the solution was stirred under Ar at -78 °C for 10 minutes and then the bath was removed. The reaction was allowed to warm up to rt and quenched by addition of NaHC0₃(_Sat.). The solution was diluted with EtOAc, washed with NaHC0₃(sa ), NaCl(sa ), dried over MgS04, filtered, concentrated to yield ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(3- fluorooxetan-3-yl)phenyl)thiazole-4-carboxylate in 99% yield. LC/MS = 459.2 (MH+), R, = 1.17 min.

Synthesis of 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(3-fluorooxetan-3- yl)phenyl)thiazole-4-carboxylic acid

Method 2 was followed using ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(3- fluorooxetan-3-yl)phenyl)thiazole-4-carboxylate to give 5-((tert-butoxycarbonyl)amino)- 2-(2,6-difluoro-4-(3-fluorooxetan-3-yl)phenyl)thiazole-4-carboxylic acid 73% yield. LC/MS = 430.0 (MH⁺), R, = 0.98 min.

Synthesis of 4-(3,5-difluorophenyl)tetrahvdro-2H-pyran-4-ol

To a solution of l-bromo-3,5-difluorobenzene in THF (0.16 M) under N₂ was added Mg turnings (1.6 equiv.). A reflux condenser was attached and the solution was submerged in a 90 °C oil bath and refluxed for 2 hours at which time the heat was removed and the solution cooled to 0°C. Dihydro-2H-pyran-4(3H)-one (1.0 equiv.) in THF was added and the solution was stirred under N₂ allowing to warm to rt for 16 hrs. The reaction was quenched by addition of sat. NH₄C1 and the solution was extracted with EtOAc, washed with brine, dried over sodium sulfate, filtered, concentrated. The crude material was purified by ISCO Si0₂ chromatography eluting with 0-100% EtOAc/n-heptanes to yield 4-(3,5-difluorophenyl)tetrahydro-2H-pyran-4-ol in 37% yield. 1H NMR (400 MHz, <cdcl3>) 5 ppm 1.63 (d, J=12.13 Hz, 2 H), 2.11 (ddd, J=13.50, 11.15, 6.65 Hz, 2 H), 3.84 - 3.90 (m, 4 H), 6.72 (tt, J=8.75, 2.20 Hz, 1 H), 6.97 - 7.05 (m, 2 H).

Synthesis of 4-(3,5-difluorophenyl)-3,6-dihvdro-2H-pyran

4-(3,5-difluorophenyl)tetrahydro-2H-pyran-4-ol (1.0 equiv.) was dissolved in DCM (0.2 M) and cooled to 0 °C. TEA (2.8 equiv.) was added to the solution, followed by MsCl (1.3 equiv.). The reaction was stirred at rt for 2hrs. The solution was cooled to 0°C and DBU (3.0 equiv.) was added. The reaction was stirred at rt for 18hrs. The solution was concentrated and the residue was purified by Si0₂ chromatography (0-100%) EtOAc in Heptanes) to afford 4-(3,5-difluorophenyl)-3,6-dihydro-2H-pyran in 38% yield. 1H NMR (400 MHz, <cdcl3>) δ ppm 2.42 - 2.49 (m, 2 H), 3.93 (t, J=5.48 Hz, 2 H), 4.32 (q, J=2.74 Hz, 2 H), 6.16 - 6.22 (m, 1 H), 6.70 (tt, J=8.80, 2.35 Hz, 1 H), 6.85 - 6.94 (m, 2 H).

Synthesis of 4-(3,5-difluorophenyl)tetrahvdro-2H-pyran

a solution of 4-(3,5-difluorophenyl)-3,6-dihydro-2H-pyran (1.0 equiv.) in methanol 2 M) was added 10%> Pd/C (0.05 equiv.). The reaction was placed under an atmosphere of hydrogen and stirred for 18 hours. Upon completion, the solution was filtered over a pad of Celite, the pad was washed with DCM, the filtrate was concentrated in vacuo to give 4-(3,5-difluorophenyl)tetrahydro-2H-pyran in 71% yield. 1H NMR (400 MHz, <cdcl3>) δ ppm 1.76 (br. s., 4 H), 2.75 (br. s., 1 H), 3.50 (br. s., 2 H), 4.08 (d, J=9.78 Hz, 2 H), 6.56 - 6.94 (m, 3 H).

Synthesis of 2-(2,6-difluoro-4-(tetrahydro-2H-pyran-4-yl)phenyl)-4,4,5,5- tetramethyl- 1 ,3 ,2-dioxaborolane

Method 3 was followed using 2-isopropoxy-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (2.2 equiv.), butyllithium (1.1 equiv.) and 4-(3,5-difluorophenyl)tetrahydro-2H-pyran (1.0 equiv.) to give 2-(2,6-difluoro-4-(tetrahydro-2H-pyran-4-yl)phenyl)-4,4,5,5-tetramethyl-

1,3,2-dioxaborolane in 100% yield. ^!H NMR (400 MHz, <cdcl3>) δ ppm 1.16 - 1.19 (m, 12 H), 1.65 - 1.74 (m, 4 H), 2.60 - 2.75 (m, 1 H), 3.37 - 3.51 (m, 2 H), 4.01 (dt, J=11.54, 3.42 Hz, 2 H), 6.67 (d, J=8.22 Hz, 2 H)

Synthesis of ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(tetrahydro-2H- pyran-4-yl)phenyl)thiazole-4-carboxylate

Method 1 was followed using ethyl 2-bromo-5-((tert-butoxycarbonyl)amino)thiazole-4- carboxylate (1.0 equiv.) and 2-(2,6-difluoro-4-(tetrahydro-2H-pyran-4-yl)phenyl)-4,4,5,5- tetramethyl-l,3,2-dioxaborolane (2.0 equiv.) at 100 ⁰ C for 20 min under microwave irradiation to give ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(tetrahydro-2H- pyran-4-yl)phenyl)thiazole-4-carboxylate in 84% yield. LC/MS = 469.2 (MH⁺), R, = 1.20 min. 1H NMR (400 MHz, <cdcl3>) δ ppm 0.83 - 0.92 (m, 1 H) 1.21 - 1.25 (m, 3 H) 1.25 - 1.32 (m, 4 H) 1.45 (t, J=7.04 Hz, 3 H) 1.50 - 1.60 (m, 9 H) 1.65 - 1.88 (m, 5 H) 2.69 - 2.87 (m, 1 H) 3.41 - 3.62 (m, 2 H) 4.02 - 4.17 (m, 2 H) 4.40 - 4.55 (m, 2 H) 6.79 - 6.94 (m, 2 H) 9.96 - 10.08 (m, 1 H)

Synthesis of 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(tetrahvdro-2H-pyran-4- yl)phenyl)thiazole-4-carboxylic acid

Method 2 was followed using ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4- (tetrahydro-2H-pyran-4-yl)phenyl)thiazole-4-carboxylate to give 5-((tert- butoxycarbonyl)amino)-2-(2,6-difluoro-4-(tetrahydro-2H-pyran-4-yl)phenyl)thiazole-4- carboxylic acid in 72% yield. LC/MS = 441.1 (MH⁺), R, = 1.02 min. 1H NMR (400 MHz, <dmso>) δ ppm 1.01 - 1.06 (m, 4 H) 1.39 - 1.54 (m, 9 H) 1.59 - 1.76 (m, 5 H) 2.78 - 2.93 (m, 1 H) 3.34 - 3.57 (m, 5 H) 3.86 - 3.96 (m, 2 H) 7.21 (d, J=9.78 Hz, 2 H) 10.06 (s, 1 H)

Synthesis of 4-(3,5-difluorophenyl)tetrahydro-2H-pyran-4-ol

To a solution of l-bromo-3,5-difluorobenzene (1.6 equiv.) in THF (0.26 M) under Ar was added Mg turnings (1.6 equiv.). A reflux condenser was attached and the solution was submerged in a 90 °C oil bath and refluxed for two hours. The dihydro-2H-pyran-4(3H)- one (1.0 equiv.) was added in THF via syringe. The solution was left stirring at rt under Ar for 5 hrs. The reaction solution was quenched by addition of NH₄Cl_(sat) and the solution was extracted with EtOAc, washed with NaCl_{(sa )}, dried over MgS0₄, filtered, concentrated and purified by ISCO Si0₂ chromatography (0-100% EtOAc/n-heptanes gradient) to yield 4-(3,5-difluorophenyl)tetrahydro-2H-pyran-4-ol in 71% yield. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 1.59 - 1.68 (m, 3 H), 2.07 - 2.19 (m, 2 H), 3.87 - 3.93 (m, 4 H), 6.72 (tt, J=8.75, 2.20 Hz, 1 H), 6.97 - 7.06 (m, 2 H).

Synthesis of 4-(3,5-difluoro-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)phenyl)tetrahydro-2H-pyran-4-ol

Method 3 was followed using 2-isopropoxy-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (2.5 equiv.), butyllithium (2.4 equiv.) and 4-(3,5-difluorophenyl)tetrahydro-2H-pyran-4-ol (1.0 equiv.) to give 4-(3,5-difluoro-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)phenyl)tetrahydro-2H-pyran-4-ol in 97% yield. 1H NMR (400 MHz, <cdcl3>) δ ppm 1.32 - 1.42 (m, 12 H), 1.56 - 1.65 (m, 2 H), 2.11 (d, J=3.13 Hz, 2 H), 3.86 - 3.92 (m, 4 H), 6.99 (d, J=9.00 Hz, 2 H).

Synthesis of ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(4- hydroxytetrahydro-2H-pyran-4-yl)phenyl)thiazole-4-carboxylate

Method 1 was followed using methyl ethyl 2-bromo-5-((tert- butoxycarbonyl)amino)thiazole-4-carboxylate (1.0 equiv.) and 4-(3,5-difluoro-4-(4,4,5,5- tetramethyl-l ,3,2-dioxaborolan-2-yl)phenyl)tetrahydro-2H-pyran-4-ol (2.0 equiv.) at 100 °C for 20 min under microwave irradiation to give ethyl 5-((tert- butoxycarbonyl)amino)-2-(2,6-difluoro-4-(4-hydroxytetrahydro-2H-pyran-4- yl)phenyl)thiazole-4-carboxylate in 70% yield. LC/MS = 485.1 (MH⁺), R, = 1.07 min. 1H NMR (400 MHz, <cdcl3>) δ ppm 0.88 (t, J=6.85 Hz, 2 H) 1.20 - 1.25 (m, 2 H) 1.38 - 1.49 (m, 3 H) 1.51 - 1.58 (m, 9 H) 1.65 (d, J=13.30 Hz, 3H) 2.07 - 2.20 (m, 2 H) 3.83 - 3.96 (m, 4 H) 4.39 - 4.54 (m, 2 H) 7.16 (d, J=9.78 Hz, 2 H) 10.03 (s, 1 H)

Synthesis of 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(4-hvdroxytetrahydro-2H- pyran-4-yl)phenyl)thiazole-4-carboxylic acid

Method 2 was followed using ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(4- hydroxytetrahydro-2H-pyran-4-yl)phenyl)thiazole-4-carboxylate to give 5-((tert- butoxycarbonyl)amino)-2-(2,6-difluoro-4-(4-hydroxytetrahydro-2H-pyran-4- yl)phenyl)thiazole-4-carboxylic acid in 86% yield. LC/MS = 457.0 (MH⁺), R, = 0.86 min. 1H NMR (400 MHz, <dmso>) δ ppm 1.42 - 1.55 (m, 13 H) 1.90 - 2.07 (m, 3 H) 3.60 - 3.81 (m, 6 H) 7.36 (d, J=10.56 Hz, 2 H) 10.07 (s, 1 H)

Synthesis of ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(4- fluorotetrahvdro-2H-pyran-4-yl)phenyl)thiazole-4-carboxylate

To a solution of ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(4- hydroxytetrahydro-2H-pyran-4-yl)phenyl)thiazole-4-carboxylate (1.0 equiv.) in CH₂CI₂ (0.01 M) at -78 °C was added DASTF (1.0 equiv.) dropwise. The resulting mixture was allowed to warm to RT and stirred at this temperature for a further 2 hrs. The reaction mixture was then quenched with NaHC0₃ and diluted with EtOAc. The aqueous layer was separated then extracted with EtOAc. The combined organics were dried over MgS04 and concentrated in vacuo to yield ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6- difluoro-4-(4-fluorotetrahydro-2H-pyran-4-yl)phenyl)thiazole-4-carboxylate in 100% yield. LC/MS = 487.1 (MH+), Rt = 1.21 min. The product was utilized in the subsequent reaction without further purification. Synthesis of 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(4-fluorotetrahvdro-2H- pyran-4-yl)phenyl)thiazole-4-carboxylic acid

Method 2 was followed using ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(4- fluorotetrahydro-2H-pyran-4-yl)phenyl)thiazole-4-carboxylate to give 5-((tert- butoxycarbonyl)amino)-2-(2,6-difluoro-4-(4-fluorotetrahydro-2H-pyran-4- yl)phenyl)thiazole-4-carboxylic acid in 62% yield. LC/MS = 459.0 (MH⁺), R, = 1.01 min. Synthesis of ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difluoro-4-(tetrahvdro-2H- pyran-4-yl)phenyl)thiazole-4-carboxylate

Method 1 was followed using ethyl 2-bromo-5-(tert-butoxycarbonylamino)thiazole-4- carboxylate (1.0 equiv.) and 2-(2,6-difluoro-4-(tetrahydro-2H-pyran-4-yl)phenyl)- 4,4,5,5-tetramethyl-l,3,2-dioxaborolane (2.0 equiv.) at 100 °C for 20 min in microwave to give ethyl 5-(tert-butoxycarbonylamino)-2-(2,6-difluoro-4-(tetrahydro-2H-pyran-4- yl)phenyl)thiazole-4-carboxylate in 84% yield. LC/MS = 469.2 (MH+ ), Rt = 1.21 min. 1H NMR (400 MHz, <cdcl3>) δ ppm 0.83 - 0.92 (m, 1 H) 1.21 - 1.25 (m, 3 H) 1.25 - 1.32 (m, 4 H) 1.45 (t, J=7.04 Hz, 3 H) 1.50 - 1.60 (m, 9 H) 1.65 - 1.88 (m, 5 H) 2.69 - 2.87 (m, 1 H) 3.41 - 3.62 (m, 2 H) 4.02 - 4.17 (m, 2 H) 4.40 - 4.55 (m, 2 H) 6.79 - 6.94 (m, 2 H) 9.96 - 10.08 (m, 1 H)

Method 2 was followed using ethyl 5-((tert-butoxycarbonyl)amino)-2-(2,6-difiuoro-4- (tetrahydro-2H-pyran-4-yl)phenyl)thiazole-4-carboxylate to give 5-((tert- butoxycarbonyl)amino)-2-(2,6-difiuoro-4-(tetrahydro-2H-pyran-4-yl)phenyl)thiazole-4- carboxylic acid in 72% yield. LC/MS = 441.1 (MH+), Rt = 1.02 min. 1H NMR (400 MHz, <dmso>) δ ppm 1.01 - 1.06 (m, 4 H) 1.39 - 1.54 (m, 9 H) 1.59 - 1.76 (m, 5 H) 2.78 - 2.93 (m, 1 H) 3.34 - 3.57 (m, 5 H) 3.86 -3.96 (m, 2 H) 7.21 (d, J=9.78 Hz, 2 H) 10.06 (s, 1 H)

Synthesis of l-(3,5-difluorophenyl)cyclobutanol

To a solution of l-bromo-3,5-difiuorobenzene (1.0 equiv.) in THF (0.26 M) under added Mg turnings (1.6 equiv.). A reflux condenser was attached and the solution was submerged in a 90 °C oil bath and refluxed for two hours. The oxetan-3-one (1.0 equiv.) was added in THF via syringe. The solution was left stirring at rt under Ar for 5 hrs. The reaction solution was quenched by addition of NH₄Cl(_sat) and the solution was extracted with EtOAc, washed with NaCl(sat), dried over MgS04, filtered, concentrated and purified by ISCO Si0₂ chromatography (0-100% EtOAc/n-heptanes gradient) to yield l-(3,5-difluorophenyl)cyclobutanol in 54% yield. 1H NMR (400 MHz, CHLOROFORM- d) δ ppm 1.69 - 1.83 (m, 1 H), 2.03 - 2.13 (m, 1 H), 2.31 - 2.43 (m, 2 H), 2.45 - 2.56 (m, 2 H), 6.71 (tt, J=8.80, 2.35 Hz, 1 H), 6.98 - 7.07 (m, 2 H). Synthesis of l-(3,5-difluoro-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- vDphenvDcyclobutanol

Method 3 was followed using 2-isopropoxy-4,4,5,5-tetramethyl-l,3,2- dioxaborolane (2.5 equiv.), butyllithium (2.4 equiv.) and l-(3,5- difluorophenyl)cyclobutanol (1.0 equiv.) to give l-(3,5-difluoro-4-(4,4,5,5-tetramethyl- l,3,2-dioxaborolan-2-yl)phenyl)cyclobutanol in 100% yield. 1H NMR (400 MHz, <cdcl3>) δ ppm 1.23 - 1.25 (m, 12 H), 1.69 - 1.82 (m, 1 H), 2.05 - 2.12 (m, 1 H), 2.37 (br. s., 2 H), 2.47 (br. s., 2 H), 7.00 (d, J=8.80 Hz, 2 H). Synthesis of tert-butyl ((l S,3R,5S)-3-(3-(2-bromothiazole-4-carboxamido)pyridin-4-yl)-

5 -methylcyclohexyDcarbamate

A solution of tert-butyl ((l S,3R,5S)-3-(3-aminopyridin-4-yl)-5- methylcyclohexyl)carbamate (1.0 eq.), 2-bromothiazole-4-carboxylic acid (1.2 eq.), HO AT (1.2 eq.) and EDC (1.20) in DMF (0.2 M) was stirred at RT for 17 h. The solution was then diluted with EtOAc and washed subsequenetly with water, IN NaOH then saturated brine solution. The organic phase was dried with magnesium sulfate and concentrated in vaccuo to yield the product tert-butyl ((l S,3R,5S)-3-(3-(2-bromothiazole- 4-carboxamido)pyridin-4-yl)-5-methylcyclohexyl)carbamate as a pale yellow oil which was utilised without further purification. LC/MS = 495.0/497.0 (MH+), Rt = 0.90 min.

Synthesis of N-(4-((lR,3S,5S -3-amino-5-methylcvclohexynpyridin-3-vn-2-(2,6- difluoro-4-(4-hydroxytetrahydro-2H-pyran-4-yl)phenyl)thiazole-4-carboxamide

Method 1 was followed using tert-butyl ((l S,3R,5S)-3-(3-(2-bromothiazole-4- carboxamido)pyridin-4-yl)-5-methylcyclohexyl)carbamate (1.0 equiv.) and 2-(2,6- difluoro-4-(tetrahydro-2H-pyran-4-yl)phenyl)-4,4,5,5-tetramethyl-l ,3,2-dioxaborolane (2.0 equiv.) at 100 °C for 20 min in microwave to give N-(4-((lR,3S,5S)-3-amino-5- methylcyclohexyl)pyridin-3-yl)-2-(2,6-difluoro-4-(4-hydroxytetrahydro-2H-pyran-4- yl)phenyl)thiazole-4-carboxamide in 3% yield. LC/MS = 529.1 (MH+ ), Rt = 0.57 min. Synthesis of N-(4-((lR,3S,5S -3-amino-5-methylcvclohexynpyridin-3-vn-2-(2,6- difluoro-4-(3 -hydroxyoxetan-3 -yl)phenyl)thiazole-4-carboxamide

Method 1 was followed using tert-butyl ((lS,3R,5S)-3-(3-(2-bromothiazole-4- carboxamido)pyridin-4-yl)-5-methylcyclohexyl)carbamate (1.0 equiv.) and 3-(3,5- difluoro-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)phenyl)oxetan-3-ol (2.0 equiv.) at 100 °C for 20 min in microwave to give N-(4-((lR,3S,5S)-3-amino-5- methylcyclohexyl)pyridin-3-yl)-2-(2,6-difluoro-4-(3-hydroxyoxetan-3-yl)phenyl)thiazole- 4-carboxamide in 5% yield. LC/MS = 501.1 (MH+ ), Rt = 0.55 min.

Synthesis of N-(4-((lR,3S,5S -3-amino-5-methylcvclohexynpyridin-3-vn-2-(2,6- difiuoro-4-( 1 -hvdroxycvclobutyl)phenyl)thiazole-4-carboxamide

Method 1 was followed using tert-butyl ((lS,3R,5S)-3-(3-(2-bromothiazole-4- carboxamido)pyridin-4-yl)-5-methylcyclohexyl)carbamate (1.0 equiv.) and 1 -(3,5- difluoro-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)phenyl)cyclobutanol (2.0 equiv.) at 100 °C for 20 min in microwave to give N-(4-((lR,3S,5S)-3-amino-5- methylcyclohexyl)pyridin-3-yl)-2-(2,6-difluoro-4-(l-hydroxycyclobutyl)phenyl)thiazole- 4-carboxamide in 5% yield. LC/MS = 499.1 (MH+ ), Rt = 0.62 min.

Method 4

A homogeneous solution of 1 eq each of amine, carboxylic acid, HO AT and EDC in DMF, at a concentration of 0.5 M, was left standing for 24 hours at which time water and ethyl acetate were added. The organic phase was dried with sodium sulfate and purified via silica gel column chromatography eluting with ethyl acetate and hexanes to give the desired protected amide product. Alternatively the crude reaction mixture was directly purified by HPLC. Upon lyophilization, the TFA salt of the protected amide product was obtained. Alternatively, the HPLC fractions could be added to EtOAc and solid Na₂C03, separated and washed with NaCl(_sat). Upon drying over MgSC^, filtering and removing the volatiles in vacuo, the protected amide product was obtained as a free base. Alternatively, the crude reaction mixture was used for the deprotection step without further purification.

If an N-Boc protected amine was present, it was removed by treating with excess 4M HCl/ dioxane for 14 hours or by treating with 25% TFA/CH₂C1₂ for 2 hours. Upon removal of the volatiles in vacuo, the material was purified by RP HPLC yielding after lyophilization the amide product as the TFA salt. Alternatively, the HPLC fractions could be added to EtOAc and solid Na₂C0₃, separated and washed with NaCl_{(sa )}. Upon drying over MgS0₄, filtering and removing the volatiles in vacuo the free base was obtained. Upon dissolving in MeCN/H₂0, adding 1 eq. of 1 N HCl and lyophilizing, the HCl salt of the amide product was obtained.

If an N-Boc, OAc group were present, prior to Boc deprotection, the acetate group could be cleaved by treating with K₂C0₃ (2.0 equiv.) in ethanol at a concentration of 0.1 M for 24 hours. If a TBDMS ether was present, it was deprotected prior to Boc removal by treating with 6N HCl, THF, methanol (1 :2: 1) at room temperature for 12 h. After removal of volatiles in vacuo, the Boc amino group was deprotected as described above. Alternatively, the TBDMS ether and Boc group could be both deprotected with 6N HCl, THF, methanol (1 :2: 1) if left at rt for 24 hours, or heated at 60 °C for 3 hours.

Following the procedures of Method 4, the following compounds were prepared:

TABLE 1

In addition to LC/MS and LC characterization, representative compounds were analyzed by !H-NMR. The following data in Table 2 are typical spectra for representative compounds of the invention.

Table 2

1 H NMR (400 MHz, <cd3od>) d ppm 1.06 (d, J=6.26 Hz, 3 H) 1 .09 -

1.31 (m, 2 H) 1.54 (q, J=1 1.87 Hz, 1 H) 1.78 (br. s, 0 H) 1.95 (d, J=12.13 Hz, 1 H) 2.09 (d, J=12.13 Hz, 1 H) 2.21 (d, J=1 1.74 Hz, 1 H) 3.15 (t, J=12.13 Hz, 1 H) 3.71 - 3.77 (m, 2 H) 4.17 (dd, J=5.09, 3.52 Hz, 2 H) 6.77 (d,J=10.56 Hz, 2 H) 7.68 (d, J=5.87 Hz, 1 H) 7.68 (d, J=5.87 Hz, 1 H) 8.44 (d, J=5.87 Hz, 1 H) 9.22 (s, 1 H)

1 H NMR (400 MHz, <cd3od>) d ppm 1 .12 (d, J=6.26 Hz, 3 H) 1.47 (q, J=12.39 Hz, 1 H) 1 .73 (d, J=12.13 Hz, 2 H) 1.98 (dd, J=13.30, 2.74 Hz, 1 H) 2.24 (d, J=9.78 Hz, 1 H) 3.07 - 3.24 (m, 4 H) 3.41 (s, 3 H) 3.74 (dd,

J=5.48, 3.52 Hz, 2 H) 4.17 (m, J=9.00 Hz, 2 H) 6.79 (d, J=10.96 Hz, 2 H) 7.67 (d, J=5.48 Hz, 1 H) 8.43 (d, J=5.48 Hz, 1 H) 9.24 (s, 1 H)

1 H NMR (400 MHz, <dmso>) d ppm 0.84 (d, J=6.65 Hz, 3 H) 1.75 (d, J=5.09 Hz, 1 H) 2.65 (t, J=12.33 Hz, 1 H) 2.88 - 3.20 (m, 3 H) 3.35 - 3.64 (m, 3 H) 3.78 (d, J=9.78 Hz, 1 H) 3.83 (s, 3 H) 6.89 (d, J=10.56 Hz, 2 H) 7.31 (d, J=6.65 Hz, 1 H) 7.61 (br. s., 2 H) 8.05 (br. s., 3 H) 8.33 (d,J=6.26 Hz, 1 H) 9.03 (s, 1 H) 9.29 (s, 1 H)

1 H NMR (400 MHz, <cd3od>) d ppm 1.03 - 1 .1 1 (m, 3 H) 1 .12 - 1.33 (m, 2 H) 1.52 - 1 .66 (m, 1 H) 1.77 - 1 .92 (m, 1 H) 1.93 - 2.03 (m, 1 H) 2.08 - 2.17 (m, 1 H) 2.19 - 2.29 (m, 1 H) 3.1 1 - 3.25 (m, 1 H) 3.35 - 3.43 (m, 1 H) 4.71 - 4.78 (m, 2 H) 4.91 - 4.95 (m, 2 H) 7.42 - 7.52 (m, 2 H) 7.68 - 7.77 (m, 1 H) 8.44 - 8.51 (m, 1 H) 9.24 - 9.32 (m, 1 H)

1 H NMR (400 MHz, <cd3od>) d ppm 1 .00 (d, J=6.65 Hz, 3 H) 1.35 (d, J=5.87 Hz, 6 H) 1.69 - 1.84 (m, 2 H) 1.91 - 2.08 (m, 1 H) 2.71 - 2.87 (m, 1 H) 3.04 - 3.17 (m, 1 H) 3.22 - 3.27 (m, 1 H) 3.66 - 3.83 (m, 1 H) 3.92 - 4.07 (m, 1 H) 4.59 - 4.72 (m, 1 H) 6.63 - 6.79 (m, 2 H) 7.42 - 7.56 (m, 1 H) 8.24 - 8.41 (m, 1 H) 9.27 - 9.37 (m, 1 H)

H NMR (400 MHz, <cd3od>) d ppm 1.06 (d, J=6.65 Hz, 3 H) 1 .29 - 1.37 (m, 6 H) 1.45 - 1.60 (m, 2 H) 1.73 - 2.00 (m, 2 H) 2.10 (d, J=12.52 Hz, 1 H) 2.21 (d, J=1 1 .74 Hz, 1 H) 2.98 - 3.22 (m, 1 H) 4.67 (dt, J=12.03, 5.92 Hz, 1 H) 6.51 - 6.81 (m, 2 H) 7.69 (d, J=5.87 Hz, 1 H)

8.44 (d, J=5.87 Hz, 1 H) 9.24 (s, 1 H) 1 H NMR (400 MHz, <cd3od>) d ppm 1 .25 (d, J=6.26 Hz, 6 H) 1 .65 - 2.1 1 (m, 6 H) 3.10 - 3.19 (m, 2 H) 3.24 - 3.29 (m, 2 H) 3.30 - 3.40 (m, 1

27

H) 3.64 (s, 3 H) 4.53 - 4.62 (m, 1 H) 6.55 - 6.65 (m, 2 H) 7.43 - 7.48

(m, 1 H)

1 H NMR (400 MHz, <cd3od>) d ppm 1 .66 - 2.12 (m, 6 H) 3.14 - 3.17 (m, 1 H) 3.25 - 3.27 (m, 2 H) 3.31 - 3.42 (m, 2 H) 3.65 (s, 3 H) 4.63 -

28

4.69 (m, 2 H) 4.81 - 4.84 (m, 2 H) 7.32 - 7.39 (m, 2 H) 7.45 - 7.50 (m, 1

H)

31

4.69 (m, 2 H) 4.81 - 4.84 (m, 2 H) 7.32 - 7.39 (m, 2 H) 7.45 - 7.50 (m, 1

H)

1 H NMR (400 MHz, <dmso>) d ppm 1 .53 - 1 .90 (m, 4 H) 1 .91 - 2.08

32 (m, 2 H) 3.01 - 3.25 (m, 4 H) 3.65 (s, 7 H) 7.26 (t, J=8.80 Hz, 2 H) 7.44

(s, 1 H) 7.45 - 7.57 (m, 2 H) 7.78 (br. s. , 3 H) 8.67 (s, 1 H)

Piml, Pim2, Pim3 AlphaScreen Assays

Pim 1, Pim 2 & Pim 3 AlphaScreen assays using high ATP (11 - 125X ATP Km) were used to determine the biochemical activity of the inhibitors. The activity of Pim 1, Pim 2, & Pim 3 is measured using a homogeneous bead based system quantifying the amount of phosphorylated peptide substrate resulting from kinase-catalyzed phosphoryl transfer to a peptide substrate. Compounds to be tested are dissolved in 100% DMSO and directly distributed to a white 384-well plate at 0.25 μΐ per well. To start the reaction, 5 μΐ of 100 nM Bad peptide (Biotin-AGAGRSRHSSYPAGT-OH (SEQ ID NO: l)) and ATP (concentrations described below) in assay buffer (50 mM Hepes, pH=7.5, 5 mM MgCl₂, 0.05% BSA, 0.01% Tween-20, 1 mM DTT) is added to each well. This is followed by the addition of 5 μΐ/well of Pim 1 , Pim 2 or Pim 3 kinase in assay buffer (concentrations described below). Final assay concentrations (described below) are in 2.5% DMSO. The reactions are performed for ~2 hours, then stopped by the addition of 10 μΐ of 0.75 μg/ml anti-phospho Ser/Thr antibody (Cell Signaling), 10 μg/ml Protein A AlphaScreen beads (Perkin Elmer), and 10 μg/ml streptavidin coated AlphaScreen beads in stop/detection buffer (50 mM EDTA, 95 mM Tris, pH=7.5, 0.01% Tween-20). The stopped reactions are incubated overnight in the dark. The phosphorylated peptide is detected via an oxygen anion initiated chemiluminescence/fluorescence cascade using the Envision plate reader (Perkin Elmer).

Compounds of the foregoing examples were tested by the Pirn 1, Pirn 2 & Pirn 3 AlphaScreen assays and found to exhibit an IC50 values as shown in Table 3 below.

IC50, the half maximal inhibitory concentration, represents the concentration of test compound that is required for 50% inhibition of its target in vitro.

Cell Proliferation Assay

KMS11 (human myeloma cell line), were cultured in IMDM supplemented with 10%> FBS, sodium pyruvate and antibiotics. Cells were plated in the same medium at a density of 2000 cells per well into 96 well tissue culture plates, with outside wells vacant, on the day of assay.

Test compounds supplied in DMSO were diluted into DMSO at 500 times the desired final concentrations before dilution into culture media to 2 times final concentrations. Equal volumes of 2x compounds were added to the cells in 96 well plates and incubated at 37 °C for 3 days.

After 3 days plates were equilibrated to room temperature and equal volume of CellTiter-Glow Reagent (Promega) was added to the culture wells. The plates were agitated briefly and luminescent signal was measured with luminometer. The percent inhibition of the signal seen in cells treated with DMSO alone vs. cells treated with control compound was calculated and used to determine EC₅₀ values (i.e., the concentration of a test compound that is required to obtain 50% of the maximum effect in the cells) for tested compounds, as shown in Table 3.

Using the procedures of the Piml, Pim2, Pim3 AlphaScreen Assays the IC₅₀ concentrations of compounds of the previous examples were determined as shown in the Table 3.

Using the procedures of Cell Proliferation Assay, the EC50 concentrations of compounds of the examples were determined in KMS11 cells as shown in Table 3.

TABLE 3

0.00232 0.850

0.00107 0.729

0.00281 0.480

0.00440 2.338

0.00125 0.055

0.00157 0.239

0.00090 0.061

0.00189 0.436

0.00154 0.202

0.00207 5.550

8.72E-06 0.00039 0.000187 0.035

2.87E-05 0.00127 0.000995 1.945 0.000104 0.00597 0.00288 >10.000

1.87E-05 0.00059 0.000407 0.053

9.93E-06 0.00081 0.000278 0.169

1.79E-05 0.00064 0.000446 0.381

1.07E-05 0.00042 0.000322 0.031

1.25E-05 0.00188 0.000201 0.201

1.43E-05 0.00120 0.000251 3.840

3.51 E-05 0.01026 0.001 133 0.892

2.43E-05 0.00472 0.000721 >10.000

1.38E-05 0.00428 0.000199

1.22E-05 0.00350 0.000216

0.02007

0.786413 1

0.02157

1.308977 3 0.03185

35 0.618792

7

Compound structures in the tables marked as "Chiral" were prepared and tested in optically active form, having the absolute stereochemistry as shown; other compounds were prepared and tested in racemic form, and the depicted structure represents the relative stereochemistry at each chiral center.

Claims

1. A compound of Formula (I) :

₄ alkyl)-pyrazole ring;

W is H or NH₂;

Z is CH or N;

n is 1 or 2;

R¹ is H, OH or OMe;

R² is H or Me,

provided that when R¹ is H, R² is also H;

R³ is H or Ci-C₄ alkyl;

R⁴ is selected from R*, -OR*, -OCH₂CH₂OR*, -CH₂OR*, and a 4-6 membered cyclic ether optionally substituted with OH, OMe, or F, wherein each R* is independently C 1-C4 alkyl,

provided that R⁴ is not -OMe when Z is N;

or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1, wherein R³ is H.

3. The compound of claim 1, wherein R³ is Me. The compound of any of claims 1-3, wherein R² is H. The compound of any of claims 1-3, wherein R² is Me. The compound of any of claim 4, wherein R¹ is H. The compound of any of claim 5, wherein R¹ is OH.

8. The compound of claim 1, wherein Z is CH.

9. The compound of claim 1, which is of the formula IA:

11. The compound of claim 10, which is of the formula IB:

The compound of claim 9 or 11 , wherein

The compound of claim 10 or 11, wherein

The compound of claim 9 or 11, wherein represents a pyrazole of the

formula where R is methyl, ethyl or isopropyl.

15. The compound of claim 14, wherein R^N is methyl.

The compound of claim 1 , wherein represents a pyridine of the

formula

The compound of claim 1 , wherein R⁴ is a tetrahydropyranyl group of the

formula:

wherein R^T is H, OH, OMe, or F.

18. The compound of claim 1, wherein R⁴ is an oxetanyl group of the formula:

wherein R⁰ is H, OH, OMe, or F.

19. The compound of claim 1, wherein R⁴ is OMe, OEt, or OPr.

20. The compound of claim 1, wherein R⁴ is -CH₂OMe or -CH₂OEt.

21. The compound of claim 1 , which is selected from the compounds in Table 1.

22. A pharmaceutical composition comprising a compound of claim 1 and at least one pharmaceutically acceptable excipient.

23. The pharmaceutical composition of claim 22, further comprising an additional therapeutic agent.

24. The pharmaceutical composition of claim 23, wherein the additional therapeutic agent is selected from irinotecan, topotecan, gemcitabine, 5-fluorouracil, cytarabine, daunorubicin, PI3 Kinase inhibitors, mTOR inhibitors, DNA synthesis inhibitors, leucovorin, carboplatin, cisplatin, taxanes, tezacitabine, cyclophosphamide, vinca alkaloids, imatinib, anthracyclines, rituximab, and trastuzumab.

25. A method to treat a condition caused or exacerbated by excessive Pirn kinase activity, which comprises administering to a subject in need thereof an effective amount of a compound of claim 1.

26. The method of claim 25, wherein the condition is a cancer.

27. The method of claim 26, wherein the cancer is selected from carcinoma of the lungs, pancreas, thyroid, ovary, bladder, breast, prostate, or colon, melanoma, myeloid leukemia, multiple myeloma, erythroleukemia, villous colon adenoma, and osteosarcoma; or the autoimmune disorder is selected from Crohn's disease, inflammatory bowel disease, rheumatoid arthritis, and chronic inflammatory diseases. 28. A compound according to claim 1 for use in therapy.

29. Use of a compound according to claim 1 for the preparation of a medicament.

31. The compound according to claim 30, wherein the cancer is selected from carcinoma of the lungs, pancreas, thyroid, ovaries, bladder, breast, prostate or colon, melanoma, myeloid leukemia, multiple myeloma, erythro leukemia, villous colon adenoma, and osteosarcoma.

32. The compound of claim 31 , wherein the condition is an autoimmune disorder.

33. A method of treating a disease or condition mediated by PIM kinase, comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of Claims 1-25, or a pharmaceutically acceptable salt thereof.

34. The method of claim 33, wherein the disease is selected from carcinoma of the lungs, pancreas, thyroid, ovaries, bladder, breast, prostate or colon, melanoma, myeloid leukemia, multiple myeloma, erythro leukemia, villous colon adenoma, and

osteosarcoma; or the disease is an autoimmune disorder.

35. The method of claim 34, wherein the disease is an autoimmune disorder.

36. The method of claim 35, wherein the autoimmune disorder is selected from Crohn's disease, inflammatory bowel disease, rheumatoid arthritis, and chronic inflammatory diseases.