WO2016142508A1 - Composition for treating pancreatic neuroendocrine tumours - Google Patents

Composition for treating pancreatic neuroendocrine tumours Download PDF

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WO2016142508A1
WO2016142508A1 PCT/EP2016/055253 EP2016055253W WO2016142508A1 WO 2016142508 A1 WO2016142508 A1 WO 2016142508A1 EP 2016055253 W EP2016055253 W EP 2016055253W WO 2016142508 A1 WO2016142508 A1 WO 2016142508A1
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pi3k
akt
inhibitor
mtor pathway
pik
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PCT/EP2016/055253
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French (fr)
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Cécile VERCHERAT-BERTOLINI
Thomas Walter
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Centre Léon-Bérard
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7008—Compounds having an amino group directly attached to a carbon atom of the saccharide radical, e.g. D-galactosamine, ranimustine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/436—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a six-membered ring having oxygen as a ring hetero atom, e.g. rapamycin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4375—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a six-membered ring having nitrogen as a ring heteroatom, e.g. quinolizines, naphthyridines, berberine, vincamine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4745—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • composition for the treatment of pancreatic neuroendocrine tumors Composition for the treatment of pancreatic neuroendocrine tumors
  • the present invention relates to a composition for the treatment of neuroendocrine tumors, in particular pancreatic neuroendocrine tumors and more particularly to a combination of at least two active principles for the treatment of these tumors.
  • Pancreatic neuroendocrine tumors (or endocrine tumors of the pancreas or pancreatic islet tumors) are rare and heterogeneous tumors in their anatomopathological and clinical presentation. This makes diagnosis, prognosis and treatment relatively complicated. In addition, it has been shown that these tumors, whose proliferation is weak and whose response to apoptosis is deregulated, are chemoresistant. Therapeutic options are therefore limited and the first results of targeted therapy treatments show encouraging but partial responses.
  • Streptozotocin is particularly known in the treatment of TNEPs, however it has been shown the appearance of resistance to this molecule.
  • the PI3K / AKT / mTOR pathway is an intracellular signaling pathway regulating cell growth, cell proliferation, cell survival and angiogenesis.
  • This signaling pathway is widely studied in the context of oncogenesis since most of the proteins constituting it are encoded by tumor suppressor genes or proto-oncogenes whose mutation can promote the development of a tumor process.
  • This signaling pathway is in particular described by Dreyer et al (Cancero dig., 2009, vol.1, No. 3, 187-190).
  • PI3K is a kinase heterodimer composed of two proteins (a p85 regulatory subunit and a p1 catalytic subunit).
  • AKT is a protein kinase.
  • mTOR mimmalian Target Of Rapamycin
  • RAF1 or RAPT1 is a 289 kDa serine / threonine kinase of the family of PIKKs (phosphoinositide 3-kinase-like kinases).
  • mTOR associates with several other proteins to form two distinct complexes called mTOR complex 1 (mTORC-1) and mTOR complex 2 (mTORC2).
  • mTOR is a central regulator of cell growth and proliferation. More specifically, mTOR regulates cell proliferation, cell growth, cell mobility, cell survival, protein synthesis and transcription. mTOR is activated by the PI3K / Akt axis and in turn phosphorese the effectors downstream of the PI3K / Akt signaling pathway. The mTOR signaling pathway is described in particular in Zoncu et al (Nature Rev Mol Cell Biol, 201 1, 12, 21-35).
  • An object of the present invention is therefore to provide new therapeutic axes for the treatment of neuroendocrine tumors, including TNEPs to overcome resistance acquired.
  • Another object of the present invention is also to provide novel therapeutic axes for improving the treatment of chemoresistant tumors.
  • Another objective of the present invention is to provide new therapeutic axes for obtaining better results in terms of apoptosis and tumor melting (reduction of the tumor surface).
  • the subject of the present invention is therefore a composition comprising streptozotocin or a derivative thereof and at least one inhibitor of the PI3K / AKT / mTOR pathway, and their pharmaceutically acceptable salts, forms or derivatives.
  • the subject of the present invention is a composition comprising streptozotocin and at least one inhibitor of the PI3K / AKT / mTOR pathway, and their pharmaceutically acceptable salts.
  • the subject of the present invention is a composition comprising streptozotocin and at least one inhibitor of the PI3K / AKT / mTOR pathway.
  • Streptozotocin is 2-deoxy-2- ( ⁇ [methyl (nitrosoamino] carbonyl ⁇ amino) -3-D-glucopyranose (CAS RN: 18883-66-4) This compound and methods of synthesis These compounds are especially described in US Pat.
  • the term inhibitor of the PI3K / AKT / mTOR pathway means any compounds capable of blocking this signaling pathway.
  • the PI3K / AKT / mTOR signaling pathway is known to those skilled in the art and in particular described by Dreyer et al (Cancero dig., 2009, vol.1, No. 3, 187-190).
  • the term inhibitor of the PI3K / AKT / mTOR pathway means any compounds capable of inhibiting one of the mTORCI and / or mTORC2 complexes and / or one of the PI3K enzymes and / or or Akt intervening upstream of the mTOR path.
  • the aforementioned inhibitors inhibit the action of the targeted proteins.
  • the inhibitors of the PI3K / AKT / mTOR pathway are chosen from:
  • PI3K inhibitors PI3K inhibitors, mTORCI and mTORC2, alone or in mixture.
  • the inhibitors of the PI3K / AKT / mTOR pathway are chosen from:
  • Such inhibitors are especially described in the documents WO2006 / 065601, WO2007 / 090913, WO2008 / 0318947, WO2008 / 144463, WO2009 / 071888, WO2009 / 032651, WO2009 / 032652, WO2009 / 032653, US5310903, WO95 / 16691, WO2008 / 070041.
  • inhibitors of the PI3K / AKT / mTOR pathway are selected from BEZ235, BKM120, Everolimus (or RAD001), MK-2206 dichloride, Pictilisib, LY294002, CAL-101, PI-3065, HS-173, PI-103.
  • the inhibitors of the PI3K / AKT / mTOR pathway are chosen in particular from the compounds defined in the documents WO2008 / 070041, US5310903, WO95 / 16691, WO2006122806 and BKM120, incorporated herein by reference, alone as a mixture.
  • the inhibitors of the PI3K / AKT / mTOR pathway are chosen from BEZ235, BKM120, Everolimus (or RAD001), MK-2206 dichlorate, preferably BEZ235, BKM120 or RAD001, alone or as a mixture.
  • BEZ235 is the formula compound
  • BKM120 is the formula compound
  • Everolimus or RAD001 is the compound
  • MK-2206 dichlorate is the compound of formula
  • the pharmaceutically acceptable salts may especially be base addition salts, for example they are pharmaceutically acceptable salts such as sodium salts, potassium salts or calcium salts, which are obtained by using metal hydroxides. alkaline and alkaline earthy corresponding as bases.
  • addition salts with pharmaceutically acceptable bases mention may be made of the salts with amines and in particular glucamine, N-methylglucamine, ⁇ , ⁇ -dimethylglucamine, ethanolamine, morpholine and N-methylmorpholine. or lysine.
  • the salts may also be obtained with inorganic or organic acids and preferably pharmaceutically acceptable acids such as hydrochloric, phosphoric, fumaric, citric, oxalic, sulfuric, ascorbic, tartaric, maleic, mandelic, methanesulphonic, lactobionic, gluconic, glucaric acids. succinic, sulfonic or hydroxypropanesulfonic acid.
  • the weight ratio of streptozotocin to the inhibitor of the PI3K / AKT / mTOR pathway is between 100 and 500, preferably between 100 and 300.
  • compositions of the present invention are preferably pharmaceutical compositions and may further comprise at least one pharmaceutically acceptable excipient.
  • Said pharmaceutically acceptable excipients are chosen according to the pharmaceutical form and the desired mode of administration, from the usual excipients known to those skilled in the art.
  • compositions according to the invention may be presented in forms intended for parenteral, intravenous, oral, rectal, permucosal or percutaneous administration.
  • injectable solutes or suspensions or multi-dose vials in the form of naked or coated tablets, coated tablets, capsules, capsules, pills, cachets, powders, suppositories or rectal capsules. , solutions or suspensions, for percutaneous use in a polar solvent, for permselective use.
  • Suitable excipients for such administrations are in particular derivatives of cellulose or microcrystalline cellulose, alkaline earth carbonates, magnesium phosphate, starches, modified starches, lactose for solid forms.
  • cocoa butter or polyethylene glycol stearates are the preferred excipients.
  • water, aqueous solutes, physiological saline, isotonic solutes are the most conveniently used vehicles.
  • the present invention also relates to a kit comprising, on the one hand, streptozotocin, one of its derivatives or one of its pharmaceutically acceptable salts and, on the other hand, at least one inhibitor of the PI3K / AKT / mTOR or a salt thereof, pharmaceutically acceptable forms or derivatives as defined above.
  • the kit comprises, on the one hand, streptozotocin and, on the other hand, at least one inhibitor of the PI3K / AKT / mTOR pathway.
  • streptozotocin and inhibitors of the PI3K / AKT / mTOR pathway can be packaged in separate preparations each with a pharmaceutically acceptable excipient, optionally capable of being mixed especially extemporaneously.
  • the invention also relates to the use of a composition or kit according to the invention for the preparation of a medicament.
  • the invention also relates to the composition or kit according to the invention for its use as a medicament.
  • the invention also relates to the use of a composition or a kit according to the invention for the prevention or the treatment of neuro-endocrine tumors (also called endocrine tumors), in particular pancreatic neuroendocrine tumors (TNEPs). .
  • neuro-endocrine tumors also called endocrine tumors
  • TNEPs pancreatic neuroendocrine tumors
  • the invention also relates to the compositions or kit of the invention for use in the prevention or treatment of neuroendocrocrine tumors (also called endocrine tumors), including pancreatic neuroendocrine tumors (TNEPs).
  • neuroendocrocrine tumors also called endocrine tumors
  • TNEPs pancreatic neuroendocrine tumors
  • the inventors have shown that the combination of STZ and at least one inhibitor of the PI3K / AKT / mTOR pathway according to the invention had a synergistic activity on the treatment of neuroendocrine tumors. , including TNEPs.
  • the inventors have shown a synergistic action of the combination of these compounds on apoptosis and tumor melting (or reduction of the tumor surface).
  • the present invention relates in particular to compositions, in particular, pharmaceutical, or the kit of the invention for their use for the prevention or treatment of neuroendocrine tumors, in particular neuroendocrine tumors of the pancreas.
  • the present invention relates in particular to a method for preventing or treating neuroendocrine tumors, in particular neuroendocrine tumors of the pancreas, comprising administering an effective amount of a composition or kit according to the invention to a patient who needs it.
  • the present invention relates in particular to the compositions or kit of the invention for the preparation of medicaments for the treatment of neuroendocrine tumors, in particular neuroendocrine tumors of the pancreas.
  • the present invention finally relates to a composition according to the invention or kit comprising STZ and at least one inhibitor of the PI3K / AKT / mTOR pathway as defined above, or a pharmaceutically acceptable salt, form or derivative thereof, for simultaneous, separate or sequential administration to a patient in need.
  • STZ and PI3K / AKT / mTOR inhibitor can be admixed in the same preparation containing them with a pharmaceutically acceptable vehicle or excipient. They can also be packaged in separate preparations each with a pharmaceutically acceptable excipient or vehicle, able to be mixed especially extemporaneously.
  • each of the active ingredients is packaged in a clean preparation containing a pharmaceutically acceptable vehicle or excipient.
  • the dosage may vary within important limits depending on the therapeutic indication, and the route of administration, as well as the age and weight of the subject.
  • patient who needs the above treatment is defined by those skilled in the art.
  • patient is meant a human being or an animal.
  • a physician or veterinarian may identify, through clinical tests, physical examination, biological tests or diagnoses, and family and / or medical history, subjects who require such treatment.
  • sufficient amount is meant a quantity of composition according to the invention or the combination of STZ and at least one inhibitor of the PI3K / AKT / mTOR pathway of the kit according to the present invention effective for preventing or treating pathological conditions. , especially for the prevention or treatment of endocrine or neurological tumors endocrine, including neuroendocrine tumors of the pancreas.
  • the sufficient amount can be determined by those skilled in the art, by means of conventional technique and by observation of the results obtained in similar circumstances.
  • the subject In order to determine the sufficient quantity, various factors must be taken into account by those skilled in the art, in particular and without being limited thereto: the subject, its size, its age, its general state of health, the disease involved and its degree of severity; the subject's response, the type of compound, the mode of administration, the bioavailability of the composition administered, the dosage, the concomitant use of other drugs, etc.
  • the streptozotocin / PI3K / AKT / mTOR inhibitor ratio is such that streptozotocin is administered in an amount of 500 mg / day to 1500 mg / day, preferably from 750 mg / day to 1000 mg / day and the inhibitor of the PI3K / AKT / mTOR pathway is administered from 1 mg / day to 15 mg / day, preferably from 2.5 mg / day to 10 mg / day, preferably 5mg every 2 days at 10mg / day.
  • Figure 1 shows the STZ + inhibitory effects of the PI3K / AKT / mTOR pathway on mTOR pathway activation and apoptosis (cleavage of caspase-3).
  • FIGS. 2 to 4 show the intrahepatic tumor surface after treatment with Streptozotocin, the PI3K / AKT / mTOR inhibitors (Everolimus, BEZ-235, BKM-120) or the streptozotocin and PI3K / AKT / inhibitor combination.
  • mTOR (ND Not determined).
  • the MIN6 cells are seeded in 384-well plates at a density of 5000 cells per well in 25 ⁇ l medium (DMEM (Dulbecco's Modified Eagle Medium), 5mM glucose, 15% FCS (Fetal Calf Serum), 1% Penicillin / Streptomycin ( 100 IU / ml penicillin and 100 ⁇ g / ml streptomycin), 0.1% ⁇ -mercaptoethanol) in an incubator at 37 ° C under 5% CO 2 .
  • DMEM Dynamic Eagle Medium
  • 5mM glucose 5mM glucose
  • FCS Fetal Calf Serum
  • Penicillin / Streptomycin 100 IU / ml penicillin and 100 ⁇ g / ml streptomycin
  • 0.1% ⁇ -mercaptoethanol 0.1% ⁇ -mercaptoethanol
  • the medium is changed to medium containing or not containing streptozotocin or not to an inhibitor of the PI3K / AKT / mTOR pathway (everolimus, BKM-120, MK-2206, BEZ-235, Table 1), the The cells are incubated in the medium described above with or without streptozotocin additive and / or inhibitor of the PI3K / AKT / mTOR pathway. 24h after incubation in this medium, the plates are equilibrated for 30 minutes at room temperature. 25 ⁇ of reagent cell titer glo ® reagent (Promega) is added according to the manufacturer's instructions.
  • the test results are shown in Tables 2 to 4 below.
  • the delta Bliss corresponds to the difference between the theoretical value obtained by the calculation A + B-AxB where A is the response for streptozotocin alone, B is the response for the inhibitor alone, and the value obtained. Bliss sum is then calculated by adding, for each streptozotocin / inhibitor pair, all the delta Bliss obtained.
  • Bliss sum is less than 0 the response is considered antagonistic, if Bliss sum is 0 the response is considered additive and if Bliss sum is greater than 0 the response is considered synergistic (Borisy et al. Proc Natl Acad Sci US A, 2003; 100 (13) 7977-82.
  • the MIN6 cells are seeded in 6-well plates at a density of 800,000 cells per well in 2 ml of medium (RPMI (Roswell Park Memorial Institute medium) 5mM glucose, 10% FCS, 1% Hepes, 1% sodium Pyruvate, 1% Penicillin Streptomycin / streptomycin (100 IU / ml penicillin and 100 ⁇ g / ml streptomycin), 0.1% ⁇ -mercaptoethanol) in an incubator at 37 ° C under 5% CO 2 .
  • medium RPMI (Roswell Park Memorial Institute medium) 5mM glucose, 10% FCS, 1% Hepes, 1% sodium Pyruvate, 1% Penicillin Streptomycin / streptomycin (100 IU / ml penicillin and 100 ⁇ g / ml streptomycin), 0.1% ⁇ -mercaptoethanol
  • the medium is changed to medium containing or not containing streptozotocin or not to inhibitors of the PI3K / AKT / mTOR pathway (see Table 5 for the concentrations used), the cells are incubated under the previous conditions in the presence of treatments for 6 hours.
  • Table 5 Concentrations Used for Inhibition of the PI3K / AKT / mTOR Pathway and Activation of Apoptosis
  • the cells are then harvested and the proteins extracted with the RIPA lysis buffer (Santa Cruz Technology): after 30 minutes of lysis with stirring, the solutions are centrifuged for 15 minutes at 13000rpm at 4 ° C. The supernatant containing the proteins is recovered.
  • the membranes are then blocked (TBS (tris-buffered saline) -Tween0,01% - 5% milk) and then incubated with the primary antibody recognizing the protein of interest overnight at 4 ° C. (see Table 6 antibodies used) . After several washes in TBS-0.01% Tween, the membrane is incubated for 1 hour in the presence of a secondary antibody directed against the primary antibody. After several washings in TBS-Tween 0.01%, the antibody-antigen complexes are revealed in luminescence thanks to the Luminata Crescendo reagent. (Millipore) on autoradiographic films. Tubulin is used as a protein load control.
  • mice xenografts with INS-1 insulinoma cell aged 5 weeks are anesthetized with isoflurane.
  • a mini left laparotomy is performed to remove the spleen from the abdominal cavity.
  • the cell suspension (2.5 million insulinoma cells in 50 ⁇ of phosphate-buffered saline PBS (sterile) is injected into the spleen, and the abdominal wall and skin are sewn in two planes with resorbable surgical wire. They are weighed twice a week and start at 7 days after transplant and last 3 to 5 weeks (as shown in Table 7).
  • the STZ solution is prepared extemporaneously in a citrate solution prepared extemporaneously (stability about 20 minutes).
  • PI3K / AKT / mTOR inhibitor solutions (t BKM-120) are prepared in DMSO and then diluted extemporaneously at the correct concentration in NaCl or water, respectively.
  • BEZ-235 is diluted in 5% methylcellulose extemporaneously.
  • Table 7 Treatment protocol for the xenograft model.
  • the mice are treated with one or the other compound, or both in combination.
  • the control mice are injected with NaCl and gavaged with NaCl or methylcellulose.
  • the drinking water is supplemented with 10% sucrose.
  • Each of the tests is carried out on 10 animals. After 3 to 5 weeks of treatment, the mice are killed, the liver, the spleen and the pancreas are removed and fixed in buffered formalin. After at least 24 hours, the organs are included in paraffin, cut and labeled in immunohistochemistry. A lining of the livers for chromogranin A makes it possible to identify the tumor cells.
  • the tumor surface is analyzed in morphometry with the HISTOLAB software (Alphelys) and the data analyzed using the GraphPad Prism software (GraphPad Version 5.03). The results are shown in Figures 2 to 4 ( * p ⁇ 0.05, ** p ⁇ 0.01).
  • CTL represents the group of control mice that did not receive STZ injections and / or PI3K / AKT / mTOR inhibitor.

Abstract

The invention relates to a composition comprising streptozotocin and at least one inhibitor of the PI3K/AKT/mTOR pathway, and the pharmaceutically acceptable salts thereof. The invention also relates to the use of said composition for the prevention or the treatment of endocrine or neuroendocrine tumours, especially endocrine tumours of the pancreas (PNETs).

Description

Composition pour le traitement des tumeurs neuroendocrines pancréatiques  Composition for the treatment of pancreatic neuroendocrine tumors
La présente invention concerne une composition pour le traitement des tumeurs neuroendocrines, notamment tumeurs neuroendocrines pancréatiques et plus particulièrement une combinaison d'au moins deux principes actifs pour le traitement de ces tumeurs. The present invention relates to a composition for the treatment of neuroendocrine tumors, in particular pancreatic neuroendocrine tumors and more particularly to a combination of at least two active principles for the treatment of these tumors.
Les tumeurs neuroendocrines pancréatiques (TNEPs) (ou tumeurs endocrines du pancréas ou tumeurs des îlots pancréatiques) sont des tumeurs rares et hétérogènes dans leur présentation anatomopathologique et clinique. Cela rend le diagnostic, le pronostic et la prise en charge thérapeutique relativement compliqués. En outre, il a été démontré que ces tumeurs, dont la prolifération est faible et dont la réponse à l'apoptose serait dérégulée, sont chimiorésistantes. Les options thérapeutiques sont donc limitées et les premiers résultats des traitements par thérapies ciblées montrent des réponses encourageantes mais partielles. Pancreatic neuroendocrine tumors (PNEs) (or endocrine tumors of the pancreas or pancreatic islet tumors) are rare and heterogeneous tumors in their anatomopathological and clinical presentation. This makes diagnosis, prognosis and treatment relatively complicated. In addition, it has been shown that these tumors, whose proliferation is weak and whose response to apoptosis is deregulated, are chemoresistant. Therapeutic options are therefore limited and the first results of targeted therapy treatments show encouraging but partial responses.
Les molécules de chimiothérapie classiques (taxol, cisplatin) sont généralement inefficaces pour cette pathologie. Cette résistance innée s'explique sans doute par leur faible taux de prolifération ainsi que l'expression accrue de certains facteurs pouvant conférer une résistance aux chimiothérapies. Les traitements cytotoxiques actuels entraînent des stabilisations mais peu de fontes tumorales. En outre, l'évolution parfois lente de la maladie (taux de survie relative de 40% à 10 ans) implique une prise en charge thérapeutique à long terme avec des traitements successifs à forte toxicité et l'apparition de résistances acquises.  Conventional chemotherapy molecules (taxol, cisplatin) are generally ineffective for this pathology. This innate resistance is probably due to their low rate of proliferation and the increased expression of certain factors that can confer resistance to chemotherapy. Current cytotoxic treatments lead to stabilization but few tumor fonts. In addition, the sometimes slow evolution of the disease (relative survival rate of 40% at 10 years) implies a long-term therapeutic management with successive treatments with high toxicity and the appearance of acquired resistance.
La streptozotocin est notamment connue dans le traitement de TNEPs, cependant il a été montré l'apparition d'une résistance à cette molécule.  Streptozotocin is particularly known in the treatment of TNEPs, however it has been shown the appearance of resistance to this molecule.
La voie PI3K/AKT/mTOR est une voie de signalisation intracellulaire régulant la croissance cellulaire, la prolifération cellulaire, la survie cellulaire et l'angiogenèse. Cette voie de signalisation est largement étudiée dans le cadre de l'oncogenèse puisque la plupart des protéines la constituant sont codées par des gènes suppresseurs de tumeurs ou des proto-oncogènes dont la mutation peut favoriser le développement d'un processus tumoral. Cette voie de signalisation est notamment décrite par Dreyer et al (Cancéro dig., 2009, vol.1 , N°3, 187-190).  The PI3K / AKT / mTOR pathway is an intracellular signaling pathway regulating cell growth, cell proliferation, cell survival and angiogenesis. This signaling pathway is widely studied in the context of oncogenesis since most of the proteins constituting it are encoded by tumor suppressor genes or proto-oncogenes whose mutation can promote the development of a tumor process. This signaling pathway is in particular described by Dreyer et al (Cancero dig., 2009, vol.1, No. 3, 187-190).
Parmi les protéines impliquées dans la voie PI3K/AKT/mTOR on peut citer les trois protéines PI3K, AKT et mTOR. PI3K est un hétérodimère à activité kinase constitué de deux protéines (une sous-unité régulatrice p85 et une sous-unité catalytique p1 10). AKT est une protéine kinase. mTOR (mammalian Target Of Rapamycin), également connu sous le nom de FRAP, RAF1 ou RAPT1 est une sérine/thréonine kinase de 289 kDa de la famille des PIKKs (phosphoinositide 3-kinase-like kinases). mTOR s'associe à plusieurs autres protéines pour former deux complexes distincts appelés mTOR complex 1 (mTORC-1 ) et mTOR complex 2 (mTORC2). mTOR est un régulateur central de la croissance et de la prolifération cellulaire. Plus particulièrement mTOR régule la prolifération cellulaire, la croissance cellulaire, la mobilité cellulaire, la survie cellulaire, la synthèse protéique et la transcription. mTOR est activé par l'axe PI3K/Akt et à son tour phosphoryle les effecteurs en aval de la voie de signalisation PI3K/Akt. La voie de signalisation mTOR est notamment décrite dans Zoncu et al (Nature Rev Mol Cell Biol, 201 1 , 12, 21 -35). Among the proteins involved in the PI3K / AKT / mTOR pathway are the three PI3K, AKT and mTOR proteins. PI3K is a kinase heterodimer composed of two proteins (a p85 regulatory subunit and a p1 catalytic subunit). AKT is a protein kinase. mTOR (mammalian Target Of Rapamycin), also known under the name FRAP, RAF1 or RAPT1 is a 289 kDa serine / threonine kinase of the family of PIKKs (phosphoinositide 3-kinase-like kinases). mTOR associates with several other proteins to form two distinct complexes called mTOR complex 1 (mTORC-1) and mTOR complex 2 (mTORC2). mTOR is a central regulator of cell growth and proliferation. More specifically, mTOR regulates cell proliferation, cell growth, cell mobility, cell survival, protein synthesis and transcription. mTOR is activated by the PI3K / Akt axis and in turn phosphorese the effectors downstream of the PI3K / Akt signaling pathway. The mTOR signaling pathway is described in particular in Zoncu et al (Nature Rev Mol Cell Biol, 201 1, 12, 21-35).
Il a été décrit l'utilisation d'inhibiteur de la voie de signalisation mTOR pour le traitement des TNEPs. Cependant, les résultats obtenus montrent que la fonte tumorale reste limitée (réponse objective inférieure à 10%) et montrent également l'apparition de résistance.  The use of inhibitor of the mTOR signaling pathway for the treatment of TNEPs has been described. However, the results obtained show that the tumor melting remains limited (objective response less than 10%) and also show the appearance of resistance.
Il existe donc un intérêt à développer de nouveaux axes thérapeutiques pour le traitement des TNEPs permettant de s'affranchir des résistances acquises. There is therefore an interest in developing new therapeutic axes for the treatment of TNEPs making it possible to overcome resistance acquired.
Un objectif de la présente invention est par conséquent de fournir de nouveaux axes thérapeutiques pour le traitement des tumeurs neuro-endocrines, notamment TNEPs permettant de s'affranchir des résistances acquises. An object of the present invention is therefore to provide new therapeutic axes for the treatment of neuroendocrine tumors, including TNEPs to overcome resistance acquired.
Un autre objectif de la présente invention est également de fournir de nouveaux axes thérapeutiques permettant l'amélioration du traitement des tumeurs chimiorésistantes.  Another object of the present invention is also to provide novel therapeutic axes for improving the treatment of chemoresistant tumors.
Un autre objectif encore de la présente invention est de fournir de nouveaux axes thérapeutiques permettant d'obtenir de meilleurs résultats en termes d'apoptose et de fonte tumorale (réduction de la surface tumorale).  Another objective of the present invention is to provide new therapeutic axes for obtaining better results in terms of apoptosis and tumor melting (reduction of the tumor surface).
D'autres objectifs apparaîtront à la lecture de la description de l'invention qui suit. La présente invention a par conséquent pour objet une composition comprenant de la streptozotocin ou un de ses dérivés et au moins un inhibiteur de la voie PI3K/AKT/mTOR, et leurs sels, formes ou dérivés pharmaceutiquement acceptables.  Other objectives will appear on reading the description of the invention which follows. The subject of the present invention is therefore a composition comprising streptozotocin or a derivative thereof and at least one inhibitor of the PI3K / AKT / mTOR pathway, and their pharmaceutically acceptable salts, forms or derivatives.
De préférence, la présente invention a pour objet une composition comprenant de la streptozotocin et au moins un inhibiteur de la voie PI3K/AKT/mTOR, et leurs sels pharmaceutiquement acceptables. De préférence, la présente invention a pour objet une composition comprenant de la streptozotocin et au moins un inhibiteur de la voie PI3K/AKT/mTOR. Preferably, the subject of the present invention is a composition comprising streptozotocin and at least one inhibitor of the PI3K / AKT / mTOR pathway, and their pharmaceutically acceptable salts. Preferably, the subject of the present invention is a composition comprising streptozotocin and at least one inhibitor of the PI3K / AKT / mTOR pathway.
La streptozotocin (STZ) correspond au composé 2-deoxy-2- ({[methyl(nitrosoamino]carbonyl}amino)-3-D-glucopyranose (N° CAS : 18883-66-4). Ce composé et des procédés de synthèse sont notamment décrits dans US3027300 et FR2034735. Ce composé est notamment commercialisé sous le nom Zanosar® par la société Keocyt. Parmi les dérivés de la streptozotocin on peut notamment citer le composé de formule Streptozotocin (STZ) is 2-deoxy-2- ({[methyl (nitrosoamino] carbonyl} amino) -3-D-glucopyranose (CAS RN: 18883-66-4) This compound and methods of synthesis These compounds are especially described in US Pat.
Figure imgf000004_0001
décrit dans FR2092093 incorporé ici par référence.
Figure imgf000004_0001
described in FR2092093 incorporated herein by reference.
Dans le cadre de la présente invention, on entend par inhibiteur de la voie PI3K/AKT/mTOR, tous composés capables de bloquer cette voie de signalisation. La voie de signalisation PI3K/AKT/mTOR est connue de l'Homme du métier et notamment décrite par Dreyer et al (Cancéro dig., 2009, vol.1 , N°3, 187-190). Plus particulièrement, dans le cadre de la présente invention, on entend par inhibiteur de la voie PI3K/AKT/mTOR tous composés capables d'inhiber l'un des complexes mTORCI et/ou mTORC2 et/ou l'une des enzymes PI3K et/ou Akt intervenant en amont de la voie mTOR. Sans vouloir être lié par une quelconque théorie, les inhibiteurs susmentionnés inhibent l'action des protéines visées. In the context of the present invention, the term inhibitor of the PI3K / AKT / mTOR pathway means any compounds capable of blocking this signaling pathway. The PI3K / AKT / mTOR signaling pathway is known to those skilled in the art and in particular described by Dreyer et al (Cancero dig., 2009, vol.1, No. 3, 187-190). More particularly, in the context of the present invention, the term inhibitor of the PI3K / AKT / mTOR pathway means any compounds capable of inhibiting one of the mTORCI and / or mTORC2 complexes and / or one of the PI3K enzymes and / or or Akt intervening upstream of the mTOR path. Without wishing to be bound by any theory, the aforementioned inhibitors inhibit the action of the targeted proteins.
De préférence, dans le cadre de la présente invention, les inhibiteurs de la voie PI3K/AKT/mTOR sont choisis parmi : Preferably, in the context of the present invention, the inhibitors of the PI3K / AKT / mTOR pathway are chosen from:
Les inhibiteurs de mTORCI ;  MTORCI inhibitors;
Les inhibiteurs de mTORC2 ;  MTORC2 inhibitors;
Les inhibiteurs de PI3K ;  PI3K inhibitors;
Les inhibiteurs de Akt ; ou  Akt inhibitors; or
- Les inhibiteurs de PI3K, mTORCI et mTORC2, seuls ou en mélange. PI3K inhibitors, mTORCI and mTORC2, alone or in mixture.
De façon particulièrement préférée, les inhibiteurs de la voie PI3K/AKT/mTOR sont choisis parmi : In a particularly preferred manner, the inhibitors of the PI3K / AKT / mTOR pathway are chosen from:
- Les inhibiteurs de PI3K, mTORCI et mTORC2 ; ou  - PI3K inhibitors, mTORCI and mTORC2; or
Les inhibiteurs de mTORCI ,  MTORCI inhibitors,
seuls ou en mélange.  alone or in mixture.
De tels inhibiteurs sont notamment décrits dans les documents WO2006/065601 , WO2007/090913, WO2008/0318947, WO2008/144463, WO2009/071888, WO2009/032651 , WO2009/032652, WO2009/032653, US5310903, W095/16691 , WO2008/070041 , WO2006122806, incorporés ici par référence, seuls ou en mélange. Such inhibitors are especially described in the documents WO2006 / 065601, WO2007 / 090913, WO2008 / 0318947, WO2008 / 144463, WO2009 / 071888, WO2009 / 032651, WO2009 / 032652, WO2009 / 032653, US5310903, WO95 / 16691, WO2008 / 070041. WO2006122806, incorporated herein by reference, alone or in admixture.
De préférence, les inhibiteurs de la voie PI3K/AKT/mTOR sont choisis parmi BEZ235, BKM120, Everolimus (ou RAD001 ), MK-2206 dichlorate, Pictilisib, LY294002, CAL-101 , PI-3065, HS-173, PI-103, NU7441 , TGX-221 , IC-871 14, Wortmannin, XL147, ZSTK474, BYL719, AS-605240, PIK-75, 3-methyladenine, A66, SAR245409, PIK-93, GSK2126458, PIK-90, PF-04691502, AZD6482, Apitolisib, GSK1059615, Duvelisib, Gedatolisib, TG100-1 15, AS-252424, BGT226, CUDC-907, PIK-294, AS-604850, GSK2636771 , BAY80-6946, YM201636, CH5132799, CAY10505, PIK-293, PKI-402, TG100713, VS-5584, CZC24832, Quercetin, Rapamycin, AZD8055, Temsirolimus, KU- 0063794, Zotarolimus, Torkinib, Tacrolimus, Ridaforolimus, INK 128, IPI-145, SAR245409, Torinl , USI-027, WIE-354, AZD2014, Torin 2, WYE-125132, BET226, Palomid 529, PP121 , PP242 WYE-687, CH5132799, WAY-600, Chrysophanic, ETP- 46464, GDC-0349, GDC-0941 , GDC-0068 XL388, Perifosine, GSK690693, Ipatasertib, AZD5363, AT13148, PF-04691502, AT7867, Triciribine, CCT128930, A-674563, PHT- 427, Miltefosine, Honokiol, TIC10, seuls ou en mélange. Preferably, inhibitors of the PI3K / AKT / mTOR pathway are selected from BEZ235, BKM120, Everolimus (or RAD001), MK-2206 dichloride, Pictilisib, LY294002, CAL-101, PI-3065, HS-173, PI-103. , NU7441, TGX-221, IC-87114, Wortmannin, XL147, ZSTK474, BYL719, AS-605240, PIK-75, 3-methyladenine, A66, SAR245409, PIK-93, GSK2126458, PIK-90, PF-04691502, AZD6482, Apitolisib, GSK1059615, Duvelisib, Gedatolisib, TG100-115, AS-252424, BGT226, CUDC-907, PIK-294, AS-604850, GSK2636771, BAY80-6946, YM201636, CH5132799, CAY10505, PIK-293, PKI For example: AZD2014, Torin 2, WYE-125132, BET226, Palomid 529, PP121, PP242 WYE-687, CH5132799, WAY-600, Chrysophanic, ETP-46464, GDC-0349, GDC-0941, GDC-0068 XL388, Perifosine, GSK690693, Ipatasertib, AZD5363, AT13148, PF-04691502, AT7867, Triciribine, CCT128930, A-674 563, PHT-427, Miltefosine, Honokiol, TIC10, alone or in admixture.
De façon préférée, les inhibiteurs de la voie PI3K/AKT/mTOR sont notamment choisis parmi les composés définis dans les documents WO2008/070041 , US5310903, W095/16691 , WO2006122806 et BKM120, incorporés ici par référence, seuls en mélange. Preferably, the inhibitors of the PI3K / AKT / mTOR pathway are chosen in particular from the compounds defined in the documents WO2008 / 070041, US5310903, WO95 / 16691, WO2006122806 and BKM120, incorporated herein by reference, alone as a mixture.
De préférence, les inhibiteurs de la voie PI3K/AKT/mTOR sont choisis parmi BEZ235, BKM120, Everolimus (ou RAD001 ), MK-2206 dichlorate, de préférence BEZ235, BKM120 ou RAD001 , seuls ou en mélange. Preferably, the inhibitors of the PI3K / AKT / mTOR pathway are chosen from BEZ235, BKM120, Everolimus (or RAD001), MK-2206 dichlorate, preferably BEZ235, BKM120 or RAD001, alone or as a mixture.
BEZ235 est le composé de formule BEZ235 is the formula compound
Figure imgf000006_0001
Figure imgf000006_0001
BKM120 est le composé de formule  BKM120 is the formula compound
Everolimus ou RAD001 ) est le composé  Everolimus or RAD001) is the compound
Figure imgf000006_0002
Figure imgf000006_0002
MK-2206 dichlorate est le composé de formule  MK-2206 dichlorate is the compound of formula
Les sels pharmaceutiquement acceptables peuvent notamment être des sels d'addition avec des bases, ils sont par exemple des sels pharmaceutiquement acceptables tels que les sels de sodium, les sels de potassium, les sels de calcium, lesquels sont obtenus en utilisant des hydroxydes de métaux alcalins et alcalino-terreux correspondants comme bases. Comme autre type de sels d'addition avec des bases pharmaceutiquement acceptables, on peut citer les sels avec des aminés et notamment la glucamine, le N-méthylglucamine, le Ν,Ν-diméthylglucamine, l'éthanolamine, la morpholine, la N-méthylmorpholine ou la lysine. Les sels peuvent également être obtenus avec des acides minéraux ou organiques et de préférence des acides pharmaceutiquement acceptables tels que les acides chlorhydrique, phosphorique, fumarique, citrique, oxalique, sulfurique, ascorbique, tartrique, maléique, mandélique, méthanesulfonique, lactobionique, gluconique, glucarique, succinique, sulfonique ou hydroxypropanesulfonique. The pharmaceutically acceptable salts may especially be base addition salts, for example they are pharmaceutically acceptable salts such as sodium salts, potassium salts or calcium salts, which are obtained by using metal hydroxides. alkaline and alkaline earthy corresponding as bases. As another type of addition salts with pharmaceutically acceptable bases, mention may be made of the salts with amines and in particular glucamine, N-methylglucamine, Ν, Ν-dimethylglucamine, ethanolamine, morpholine and N-methylmorpholine. or lysine. The salts may also be obtained with inorganic or organic acids and preferably pharmaceutically acceptable acids such as hydrochloric, phosphoric, fumaric, citric, oxalic, sulfuric, ascorbic, tartaric, maleic, mandelic, methanesulphonic, lactobionic, gluconic, glucaric acids. succinic, sulfonic or hydroxypropanesulfonic acid.
On entend par formes ou dérivés pharmaceutiquement acceptables, des formes modifiées ou dérivées des composés ci-dessus leur permettant d'être assimilés par l'organisme.  The term pharmaceutically acceptable forms or derivatives, forms modified or derived from the above compounds allowing them to be assimilated by the body.
De préférence, dans les compositions de la présente invention, le ratio en poids de streptozotocin par rapport à l'inhibiteur de la voie PI3K/AKT/mTOR est compris entre 100 et 500, de préférence entre 100 et 300. Preferably, in the compositions of the present invention, the weight ratio of streptozotocin to the inhibitor of the PI3K / AKT / mTOR pathway is between 100 and 500, preferably between 100 and 300.
Les compositions de la présente invention, sont de préférence des compositions pharmaceutiques et peuvent en outre comprendre au moins un excipient pharmaceutiquement acceptable. The compositions of the present invention are preferably pharmaceutical compositions and may further comprise at least one pharmaceutically acceptable excipient.
Lesdits excipients pharmaceutiquement acceptables sont choisis selon la forme pharmaceutique et le mode d'administration souhaité, parmi les excipients habituels connus de l'Homme du métier.  Said pharmaceutically acceptable excipients are chosen according to the pharmaceutical form and the desired mode of administration, from the usual excipients known to those skilled in the art.
Les compositions pharmaceutiques selon l'invention peuvent être présentées sous des formes destinées à l'administration par voie parentérale, intraveineuse, orale, rectale, permuqueuse ou percutanée.  The pharmaceutical compositions according to the invention may be presented in forms intended for parenteral, intravenous, oral, rectal, permucosal or percutaneous administration.
Elles seront donc présentées sous forme de solutés ou de suspensions injectables ou flacons multi-doses, sous forme de comprimés nus ou enrobés, de dragées, de capsules, de gélules, de pilules, de cachets, de poudres, de suppositoires ou de capsules rectales, de solutions ou de suspensions, pour l'usage percutané dans un solvant polaire, pour l'usage permuqueux.  They will therefore be presented in the form of injectable solutes or suspensions or multi-dose vials, in the form of naked or coated tablets, coated tablets, capsules, capsules, pills, cachets, powders, suppositories or rectal capsules. , solutions or suspensions, for percutaneous use in a polar solvent, for permselective use.
Les excipients qui conviennent pour de telles administrations sont notamment les dérivés de la cellulose ou de la cellulose microcristalline, les carbonates alcalino-terreux, le phosphate de magnésium, les amidons, les amidons modifiés, le lactose pour les formes solides. Pour l'usage rectal, le beurre de cacao ou les stéarates de polyéthylèneglycol sont les excipients préférés. Suitable excipients for such administrations are in particular derivatives of cellulose or microcrystalline cellulose, alkaline earth carbonates, magnesium phosphate, starches, modified starches, lactose for solid forms. For rectal use, cocoa butter or polyethylene glycol stearates are the preferred excipients.
Pour l'usage parentéral, l'eau, les solutés aqueux, le sérum physiologique, les solutés isotoniques sont les véhicules les plus commodément utilisés.  For parenteral use, water, aqueous solutes, physiological saline, isotonic solutes are the most conveniently used vehicles.
La présente invention concerne également un kit comprenant d'une part de la streptozotocin, un de ses dérivés ou un de ses sels pharmaceutiquement acceptables et d'autre part au moins un inhibiteur de la voie PI3K/AKT/mTOR ou un de leurs sels, formes ou dérivés pharmaceutiquement acceptables tels que définis plus haut. De préférence, le kit comprend d'une part de la streptozotocin et d'autre part au moins un inhibiteur de la voie PI3K/AKT/mTOR. Dans ce kit, la streptozotocin et les inhibiteurs de la voie PI3K/AKT/mTOR peuvent être conditionnés dans des préparations séparées chacun avec un excipient pharmaceutiquement acceptable, éventuellement aptes à être mélangés notamment extemporanément. The present invention also relates to a kit comprising, on the one hand, streptozotocin, one of its derivatives or one of its pharmaceutically acceptable salts and, on the other hand, at least one inhibitor of the PI3K / AKT / mTOR or a salt thereof, pharmaceutically acceptable forms or derivatives as defined above. Preferably, the kit comprises, on the one hand, streptozotocin and, on the other hand, at least one inhibitor of the PI3K / AKT / mTOR pathway. In this kit, streptozotocin and inhibitors of the PI3K / AKT / mTOR pathway can be packaged in separate preparations each with a pharmaceutically acceptable excipient, optionally capable of being mixed especially extemporaneously.
L'invention concerne également l'utilisation d'une composition ou d'un kit selon l'invention pour la préparation d'un médicament. The invention also relates to the use of a composition or kit according to the invention for the preparation of a medicament.
L'invention concerne également la composition ou le kit selon l'invention pour son utilisation en tant que médicament.  The invention also relates to the composition or kit according to the invention for its use as a medicament.
L'invention concerne également l'utilisation d'une composition ou d'un kit selon l'invention pour la prévention ou le traitement de tumeurs neuro-endrocrines (également appelées tumeurs endocrines), notamment les tumeurs neuro-endocrines du pancréas (TNEPs). The invention also relates to the use of a composition or a kit according to the invention for the prevention or the treatment of neuro-endocrine tumors (also called endocrine tumors), in particular pancreatic neuroendocrine tumors (TNEPs). .
L'invention concerne également les compositions ou le kit de l'invention destinés à une utilisation pour la prévention ou le traitement de tumeurs neuro-endrocrines (également appelées tumeurs endocrines), notamment des tumeurs neuro-endocrines du pancréas (TNEPs). De façon tout à fait surprenante et avantageuse, les inventeurs ont montré que l'association de STZ et d'au moins un inhibiteur de la voie PI3K/AKT/mTOR selon l'invention avait une activité synergique sur le traitement des tumeurs neuro-endocrines, notamment les TNEPs. De façon surprenant, les inventeurs ont montré une action synergique de l'association de ces composés sur l'apoptose et la fonte tumorale (ou réduction de la surface tumorale). La présente invention concerne en particulier les compositions, notamment, pharmaceutiques, ou le kit de l'invention pour leur utilisation pour la prévention ou le traitement des tumeurs neuro-endocrines, notamment tumeurs neuro-endocrines du pancréas. The invention also relates to the compositions or kit of the invention for use in the prevention or treatment of neuroendocrocrine tumors (also called endocrine tumors), including pancreatic neuroendocrine tumors (TNEPs). Quite surprisingly and advantageously, the inventors have shown that the combination of STZ and at least one inhibitor of the PI3K / AKT / mTOR pathway according to the invention had a synergistic activity on the treatment of neuroendocrine tumors. , including TNEPs. Surprisingly, the inventors have shown a synergistic action of the combination of these compounds on apoptosis and tumor melting (or reduction of the tumor surface). The present invention relates in particular to compositions, in particular, pharmaceutical, or the kit of the invention for their use for the prevention or treatment of neuroendocrine tumors, in particular neuroendocrine tumors of the pancreas.
La présente invention concerne en particulier une méthode de prévention ou de traitement des tumeurs neuro-endocrines, notamment tumeurs neuro-endocrines du pancréas, comprenant l'administration d'une quantité efficace d'une composition ou kit selon l'invention à un patient qui en a besoin. La présente invention concerne en particulier les compositions ou kit de l'invention pour la préparation de médicaments pour le traitement des tumeurs neuro-endocrines, notamment tumeurs neuro-endocrines du pancréas.  The present invention relates in particular to a method for preventing or treating neuroendocrine tumors, in particular neuroendocrine tumors of the pancreas, comprising administering an effective amount of a composition or kit according to the invention to a patient who needs it. The present invention relates in particular to the compositions or kit of the invention for the preparation of medicaments for the treatment of neuroendocrine tumors, in particular neuroendocrine tumors of the pancreas.
La présente invention concerne enfin une composition selon l'invention ou kit comprenant de la STZ et au moins un inhibiteur de la voie PI3K/AKT/mTOR tel que défini ci-dessus, ou un de leurs sels, formes ou dérivés pharmaceutiquement acceptables, pour une administration simultanée, séparée ou séquentielle à un patient qui en a besoin. Pour une administration simultanée, la STZ et l'inhibiteur de la voie PI3K/AKT/mTOR peuvent être en mélange dans la même préparation les contenant avec un véhicule ou excipient pharmaceutiquement acceptable. Ils peuvent aussi être conditionnés dans des préparations séparées chacun avec un excipient ou véhicule pharmaceutiquement acceptable, aptes à être mélangés notamment extemporanément. En cas d'administration séparée ou séquentielle, chacun des principes actifs est conditionné dans une préparation propre contenant un véhicule ou excipient pharmaceutiquement acceptable. The present invention finally relates to a composition according to the invention or kit comprising STZ and at least one inhibitor of the PI3K / AKT / mTOR pathway as defined above, or a pharmaceutically acceptable salt, form or derivative thereof, for simultaneous, separate or sequential administration to a patient in need. For simultaneous administration, STZ and PI3K / AKT / mTOR inhibitor can be admixed in the same preparation containing them with a pharmaceutically acceptable vehicle or excipient. They can also be packaged in separate preparations each with a pharmaceutically acceptable excipient or vehicle, able to be mixed especially extemporaneously. In the case of separate or sequential administration, each of the active ingredients is packaged in a clean preparation containing a pharmaceutically acceptable vehicle or excipient.
La posologie peut varier dans des limites importantes en fonction de l'indication thérapeutique, et de la voie d'administration, ainsi que de l'âge et du poids du sujet. The dosage may vary within important limits depending on the therapeutic indication, and the route of administration, as well as the age and weight of the subject.
L'identification du patient qui a besoin du traitement ci-dessus indiqué est définie par l'Homme du métier. On entend par patient un être humain ou un animal. Un médecin ou un vétérinaire peut identifier, par l'intermédiaire de tests cliniques, d'examen physique, de tests ou diagnostics biologiques et par l'historique familial et/ou médical, les sujets qui ont besoin d'un tel traitement.  The identification of the patient who needs the above treatment is defined by those skilled in the art. By patient is meant a human being or an animal. A physician or veterinarian may identify, through clinical tests, physical examination, biological tests or diagnoses, and family and / or medical history, subjects who require such treatment.
On entend par quantité suffisante, une quantité de composition selon l'invention ou de l'association de STZ et d'au moins un inhibiteur de la voie PI3K/AKT/mTOR du kit selon la présente invention efficace pour prévenir ou traiter des conditions pathologiques, notamment pour la prévention ou le traitement des tumeurs endocrines ou neuro- endocrines, notamment les tumeurs neuro-endocrines du pancréas. La quantité suffisante peut être déterminée par l'Homme du métier, par l'intermédiaire de technique conventionnelle et par l'observation des résultats obtenus dans des circonstances analogues. Pour déterminer la quantité suffisante différents facteurs doivent être pris en compte par l'homme du métier, notamment et sans y être limité : le sujet, sa taille, son âge, son état de santé général, la maladie impliquée et son degré de sévérité ; la réponse du sujet, le type de composé, le mode d'administration, la biodisponibilité de la composition administrée, le dosage, l'utilisation concomitante d'autres médicaments, etc. De préférence, dans la composition ou le kit de l'invention, le ratio streptozotocin/inhibiteur de la voie PI3K/AKT/mTOR est tel que la streptozotocin est administrée dans une quantité de 500 mg/jour à 1500 mg/jour, de préférence de 750 mg/jour à 1000 mg/jour et l'inhibiteur de la voie PI3K/AKT/mTOR est administré de 1 mg/jour à 15 mg/jour, de préférence de 2,5 mg/jour à 10 mg/jour, de préférence de 5mg tous les 2 jours à 10mg/jour. By sufficient amount is meant a quantity of composition according to the invention or the combination of STZ and at least one inhibitor of the PI3K / AKT / mTOR pathway of the kit according to the present invention effective for preventing or treating pathological conditions. , especially for the prevention or treatment of endocrine or neurological tumors endocrine, including neuroendocrine tumors of the pancreas. The sufficient amount can be determined by those skilled in the art, by means of conventional technique and by observation of the results obtained in similar circumstances. In order to determine the sufficient quantity, various factors must be taken into account by those skilled in the art, in particular and without being limited thereto: the subject, its size, its age, its general state of health, the disease involved and its degree of severity; the subject's response, the type of compound, the mode of administration, the bioavailability of the composition administered, the dosage, the concomitant use of other drugs, etc. Preferably, in the composition or kit of the invention, the streptozotocin / PI3K / AKT / mTOR inhibitor ratio is such that streptozotocin is administered in an amount of 500 mg / day to 1500 mg / day, preferably from 750 mg / day to 1000 mg / day and the inhibitor of the PI3K / AKT / mTOR pathway is administered from 1 mg / day to 15 mg / day, preferably from 2.5 mg / day to 10 mg / day, preferably 5mg every 2 days at 10mg / day.
La présente invention va maintenant être détaillée à l'aide d'exemples non limitatifs. The present invention will now be detailed using non-limiting examples.
La figure 1 représente les effets de combinaison STZ+ inhibiteur de la voie PI3K/AKT/mTOR sur l'activation de la voie mTOR et l'apoptose (clivage de la caspase-3).  Figure 1 shows the STZ + inhibitory effects of the PI3K / AKT / mTOR pathway on mTOR pathway activation and apoptosis (cleavage of caspase-3).
Les figures 2 à 4 représentent la surface tumorale intrahépatique après traitement par la Streptozotocin, les inhibiteurs de la voie PI3K/AKT/mTOR (Everolimus, BEZ-235, BKM-120) ou la combinaison Streptozotocin et inhibiteur de la voie PI3K/AKT/mTOR (ND= Non déterminé). Test d'additivité :  FIGS. 2 to 4 show the intrahepatic tumor surface after treatment with Streptozotocin, the PI3K / AKT / mTOR inhibitors (Everolimus, BEZ-235, BKM-120) or the streptozotocin and PI3K / AKT / inhibitor combination. mTOR (ND = Not determined). Additivity test:
Les cellules MIN6 sont ensemencées en plaques 384 puits à une densité de 5000 cellules par puits dans 25μΙ de milieu (DMEM (Dulbecco's Modified Eagle Médium), 5mM glucose, 15% SVF (Sérum de Veau Fœtal), 1 % de Pénicilline/Streptomycine (100 Ul/ml de pénicilline et 100 μg/ml de streptomycine), 0,1 % de β-mercaptoethanol) dans un incubateur à 37°C sous 5% de C02. 72h plus tard le milieu est changé pour du milieu contenant ou non de la Streptozotocin associée ou non à un inhibiteur de la voie PI3K/AKT/mTOR (everolimus, BKM-120, MK-2206, BEZ-235, tableau 1 ), les cellules sont mises à incuber dans le milieu décrit ci-dessus additivé ou non de streptozotocin et/ou d'inhibiteur de la voie PI3K/AKT/mTOR. 24h après incubation dans ce milieu, les plaques sont équilibrées pendant 30 minutes à température ambiante. 25μΙ de réactif cell titer glo ® reagent (Promega) est ajouté selon les instructions du constructeur. Après 2 minutes d'agitation, les plaques sont laissées à incuber 10 minutes à température ambiante et la luminométrie est lue sur un luminomètre (Luminoskan Ascent 2.5, Thermo Scientific) pendant 1 seconde. Un puits sans cellule est traité de manière similaire, le résultat obtenu pour ce contrôle négatif est retranché aux résultats obtenus pour les échantillons. Les résultats des tests sont présentés aux tableaux 2 à 4 ci-dessous. Le delta Bliss correspond à la différence entre la valeur théorique obtenue par le calcul suivant A+B-AxB où A est la réponse pour la streptozotocin seule, B est la réponse pour l'inhibiteur seul, et la valeur obtenue. Le Bliss sum est ensuite calculé par addition, pour chacun des couples streptozotocin/inhibiteur l'ensemble des delta Bliss obtenu. Si le Bliss sum est inférieur à 0 la réponse est considérée comme antagoniste, si le Bliss sum est égal à 0 la réponse est considérée comme additive et si le Bliss sum est supérieur à 0 la réponse est considérée comme synergique (Borisy et al., Proc Natl Acad Sci U S A., 2003;100(13)7977-82. The MIN6 cells are seeded in 384-well plates at a density of 5000 cells per well in 25 μl medium (DMEM (Dulbecco's Modified Eagle Medium), 5mM glucose, 15% FCS (Fetal Calf Serum), 1% Penicillin / Streptomycin ( 100 IU / ml penicillin and 100 μg / ml streptomycin), 0.1% β-mercaptoethanol) in an incubator at 37 ° C under 5% CO 2 . 72 hours later, the medium is changed to medium containing or not containing streptozotocin or not to an inhibitor of the PI3K / AKT / mTOR pathway (everolimus, BKM-120, MK-2206, BEZ-235, Table 1), the The cells are incubated in the medium described above with or without streptozotocin additive and / or inhibitor of the PI3K / AKT / mTOR pathway. 24h after incubation in this medium, the plates are equilibrated for 30 minutes at room temperature. 25μΙ of reagent cell titer glo ® reagent (Promega) is added according to the manufacturer's instructions. After 2 minutes The plates are allowed to incubate for 10 minutes at room temperature and the luminometry is read on a luminometer (Luminoskan Ascent 2.5, Thermo Scientific) for 1 second. A cell-free well is treated similarly, the result obtained for this negative control is deducted from the results obtained for the samples. The test results are shown in Tables 2 to 4 below. The delta Bliss corresponds to the difference between the theoretical value obtained by the calculation A + B-AxB where A is the response for streptozotocin alone, B is the response for the inhibitor alone, and the value obtained. Bliss sum is then calculated by adding, for each streptozotocin / inhibitor pair, all the delta Bliss obtained. If Bliss sum is less than 0 the response is considered antagonistic, if Bliss sum is 0 the response is considered additive and if Bliss sum is greater than 0 the response is considered synergistic (Borisy et al. Proc Natl Acad Sci US A, 2003; 100 (13) 7977-82.
Figure imgf000011_0001
Figure imgf000011_0001
Tableau 1 : concentrations utilisées pour les tests d'additivité  Table 1: Concentrations used for additivity tests
Figure imgf000011_0002
Figure imgf000011_0002
Tableau 2 Delta Bliss Bliss sum Table 2 Delta Bliss Bliss sum
STZ (concentration en mM)  STZ (concentration in mM)
(%) (%)  (%) (%)
0 0,5 1 2 4 8  0 0.5 1 2 4 8
0 0 0 0 0 0 0 0 0 0 0 0 0 0
0,05 0 -3 2 12 14 1 10.05 0 -3 2 12 14 1 1
MK-2206 MK-2206
0,5 0 -1 0 9 13 10 83 (concentration  0.5 0 -1 0 9 13 10 83 (concentration
5 0 0 1 7 7 5 en μΜ)  5 0 0 1 7 7 5 in μΜ)
25 0 0 0 0 -1 -1 25 0 0 0 0 -1 -1
50 0 0 0 0 -1 -1 50 0 0 0 0 -1 -1
Tableau 3  Table 3
Tableau 3  Table 3
Figure imgf000012_0002
Figure imgf000012_0002
Tableau 4  Table 4
Les résultats des essais montrent donc une synergie de l'association STZ et inhibiteur de la voie PI3K/AKT/mTOR. Evaluation de l'inhibition de la voie mTOR et de la caspase-3 en western-blot: The results of the tests thus show a synergy of the STZ association and inhibitor of the PI3K / AKT / mTOR pathway. Evaluation of the inhibition of the mTOR pathway and caspase-3 in western blot:
Les cellules MIN6 sont ensemencées en plaques 6 puits à une densité de 800 000 cellules par puits dans 2ml de milieu (RPMI (Roswell Park Mémorial Institute médium) 5mM glucose, 10% SVF, 1 % Hepes, 1 % sodium Pyruvate, 1 % Pénicilline/Streptomycine Streptomycine (100 Ul/ml de pénicilline et 100 μg/ml de streptomycine), 0,1 % de β- mercaptoethanol) dans un incubateur à 37°C sous 5% de C02. The MIN6 cells are seeded in 6-well plates at a density of 800,000 cells per well in 2 ml of medium (RPMI (Roswell Park Memorial Institute medium) 5mM glucose, 10% FCS, 1% Hepes, 1% sodium Pyruvate, 1% Penicillin Streptomycin / streptomycin (100 IU / ml penicillin and 100 μg / ml streptomycin), 0.1% β-mercaptoethanol) in an incubator at 37 ° C under 5% CO 2 .
72h plus tard le milieu est changé pour du milieu contenant ou non de la Streptozotocin associée ou non à un inhibiteurs de la voie PI3K/AKT/mTOR (voir tableau 5 pour les concentrations utilisées), les cellules sont mises à incuber dans les conditions précédentes en présence des traitements pendant 6 heures.  72 hours later the medium is changed to medium containing or not containing streptozotocin or not to inhibitors of the PI3K / AKT / mTOR pathway (see Table 5 for the concentrations used), the cells are incubated under the previous conditions in the presence of treatments for 6 hours.
Figure imgf000013_0001
Figure imgf000013_0001
Tableau 5 : concentrations utilisées pour l'inhibition de la voie PI3K/AKT/mTOR et l'activation de l'apoptose Les cellules sont ensuite récoltées et les protéines extraites avec le tampon de lyse RIPA (Santa Cruz Technology) : après 30 minutes de lyse sous agitation, les solutions sont centrifugées 15 minutes à 13000rpm à 4°C. Le surnageant contenant les protéines est récupéré.  Table 5: Concentrations Used for Inhibition of the PI3K / AKT / mTOR Pathway and Activation of Apoptosis The cells are then harvested and the proteins extracted with the RIPA lysis buffer (Santa Cruz Technology): after 30 minutes of lysis with stirring, the solutions are centrifuged for 15 minutes at 13000rpm at 4 ° C. The supernatant containing the proteins is recovered.
20μg de protéines dans du XT Sample Buffer 4X (BioRad) sont dénaturés à 95°C pendant 5 minutes puis déposés sur un gel d'acrylamide Criterion XT Bis-Tris Gel, 12%, (BioRad). Les protéines migrent dans du tampon MOPS (BioRad) à 80Volts pendant 30 minutes puis à 160Volts pendant 1 h30. Elles sont ensuite transférées sur une membrane PVDF (polyvinylidene difluoride) (immobilon, Millipore) dans du tampon de transfert Tris-Glycine Méthanol (Tris-base 20mM, Glycine 150mM, méthanol 20%) à 100Volts pendant 1 h. Les membranes sont ensuite bloquées (TBS (tris-buffered saline)-Tween0,01 % - 5% Lait) puis incubées avec l'anticorps primaire reconnaissant la protéine d'intérêt pendant une nuit à 4°C (voir tableau 6 anticorps utilisés). Après plusieurs lavages en TBS-Tween 0,01 %, la membrane est incubée pendant 1 heure en présence d'un anticorps secondaire dirigé contre l'anticorps primaire. Après plusieurs lavages en TBS-Tween 0,01 %, les complexes anticorps-antigènes sont révélés en luminescence grâce au réactif Luminata Crescendo (Millipore) sur des films autoradiographiques. La tubuline est utilisée comme témoin de charge des protéines. 20 μg of proteins in XT Sample Buffer 4X (BioRad) are denatured at 95 ° C. for 5 minutes and then deposited on a Criterion XT Bis-Tris Gel acrylamide gel, 12%, (BioRad). Proteins migrate in MOPS buffer (BioRad) at 80 volts for 30 minutes and then at 160 volts for 1 hour 30 minutes. They are then transferred to a PVDF (polyvinylidene difluoride) membrane (Immobilon, Millipore) in Tris-Glycine Methanol transfer buffer (20mM Tris-base, 150mM Glycine, 20% methanol) at 100Volts for 1 h. The membranes are then blocked (TBS (tris-buffered saline) -Tween0,01% - 5% milk) and then incubated with the primary antibody recognizing the protein of interest overnight at 4 ° C. (see Table 6 antibodies used) . After several washes in TBS-0.01% Tween, the membrane is incubated for 1 hour in the presence of a secondary antibody directed against the primary antibody. After several washings in TBS-Tween 0.01%, the antibody-antigen complexes are revealed in luminescence thanks to the Luminata Crescendo reagent. (Millipore) on autoradiographic films. Tubulin is used as a protein load control.
Figure imgf000014_0001
Figure imgf000014_0001
Tableau 6 : Anticorps utilisés pour l'analyse en western-blot  Table 6: Antibodies Used for Western Blot Analysis
Les résultats sont représentés à la figure 1 . Ces résultats montrent, selon les inhibiteurs une inhibition de PI3K, mTOR et/ou AKT. Le BKM-120, quant à lui, entraîne une inhibition totale de la voie PI3K/AKT/mTOR. The results are shown in Figure 1. These results show, according to the inhibitors, an inhibition of PI3K, mTOR and / or AKT. As for the BKM-120, it entails a total inhibition of the PI3K / AKT / mTOR pathway.
Concernant l'activation de l'apoptose par clivage de la caspase-3, les résultats montrent une augmentation du clivage de la caspase-3 et donc de l'apoptose par rapport aux résultats obtenus pour la STZ seule. Evaluation des combinaisons in vivo Concerning the activation of apoptosis by cleavage of caspase-3, the results show an increase in cleavage of caspase-3 and thus of apoptosis compared to the results obtained for STZ alone. Evaluation of in vivo combinations
Des souris xenogreffes avec cellule d'insulinome INS-1 âgées de 5 semaines sont anesthésiées à l'isoflurane. Une mini laparotomie gauche est pratiquée pour sortir la rate de la cavité abdominale. La suspension cellulaire (2,5 millions de cellules d'insulinome dans 50μΙ de PBS phosphate-buffered saline (stérile) est injectée dans la rate. La paroi abdominale et la peau sont recousues en deux plans à l'aide de fil chirurgical résorbable. Ils sont pesés deux fois par semaine. Le traitement débute 7 jours après la greffe et dure 3 à 5 semaines (comme indiqués dans le tableau 7).  Mice xenografts with INS-1 insulinoma cell aged 5 weeks are anesthetized with isoflurane. A mini left laparotomy is performed to remove the spleen from the abdominal cavity. The cell suspension (2.5 million insulinoma cells in 50μΙ of phosphate-buffered saline PBS (sterile) is injected into the spleen, and the abdominal wall and skin are sewn in two planes with resorbable surgical wire. They are weighed twice a week and start at 7 days after transplant and last 3 to 5 weeks (as shown in Table 7).
La solution de STZ est préparée extemporanément dans une solution citrate préparée extemporanément (stabilité environ 20 minutes). Les solutions d'inhibiteur de la voie PI3K/AKT/mTOR (t BKM-120) sont préparées dans du DMSO puis diluées extemporanément à la bonne concentration dans du NaCI ou de l'eau respectivement. Le BEZ-235 est dilué dans du méthylcellulose 5% extemporanément.  The STZ solution is prepared extemporaneously in a citrate solution prepared extemporaneously (stability about 20 minutes). PI3K / AKT / mTOR inhibitor solutions (t BKM-120) are prepared in DMSO and then diluted extemporaneously at the correct concentration in NaCl or water, respectively. BEZ-235 is diluted in 5% methylcellulose extemporaneously.
Figure imgf000015_0001
Figure imgf000015_0001
IP= intrapéritonéale  IP = intraperitoneal
Tableau 7 : protocole de traitement pour le modèle de xénogreffes. Les souris sont traitées avec l'un ou l'autre des composés, ou les deux en combinaison. Les souris contrôles sont injectées avec du NaCI et gavées avec du NaCI ou du méthylcellulose.  Table 7: Treatment protocol for the xenograft model. The mice are treated with one or the other compound, or both in combination. The control mice are injected with NaCl and gavaged with NaCl or methylcellulose.
L'eau de boisson est supplémentée avec du sucrose 10%. Chacun des essais est réalisé sur 10 animaux Après 3 à 5 semaines de traitement, les souris sont mises à mort, le foie, la rate et le pancréas sont prélevés et fixés dans du formol tamponné. Après au minimum 24h, les organes sont inclus en paraffine, coupés et marqués en immunohistochimie. Un marquage des foies pour la chromogranin A permet d'identifier les cellules tumorales. La surface tumorale est analysée en morphométrie avec le logiciel HISTOLAB (Alphelys) et les données analysées à l'aide du logiciel GraphPad Prism (GraphPad Version 5.03). Les résultats sont présentés aux figures 2 à 4 (*p<0,05, **p<0,01 ). CTL représente le groupe des souris témoins n'ayant pas reçu les injections de STZ et/ou inhibiteur de la voie PI3K/AKT/mTOR. The drinking water is supplemented with 10% sucrose. Each of the tests is carried out on 10 animals. After 3 to 5 weeks of treatment, the mice are killed, the liver, the spleen and the pancreas are removed and fixed in buffered formalin. After at least 24 hours, the organs are included in paraffin, cut and labeled in immunohistochemistry. A lining of the livers for chromogranin A makes it possible to identify the tumor cells. The tumor surface is analyzed in morphometry with the HISTOLAB software (Alphelys) and the data analyzed using the GraphPad Prism software (GraphPad Version 5.03). The results are shown in Figures 2 to 4 ( * p <0.05, ** p <0.01). CTL represents the group of control mice that did not receive STZ injections and / or PI3K / AKT / mTOR inhibitor.
Ces résultats montrent que pour les combinaisons STZ+Everolimus, STZ+BKM-120 et STZ+BEZ-235, la surface tumorale intrahépatique diminue de manière significative par rapport aux souris contrôles non traitées, ce qui n'est pas le cas lors d'un traitement STZ seul, Everolimus seul ou BEZ-235 seul.  These results show that for the STZ + Everolimus, STZ + BKM-120 and STZ + BEZ-235 combinations, the intrahepatic tumor area decreases significantly compared to untreated control mice, which is not the case when STZ alone, Everolimus alone or BEZ-235 alone.

Claims

REVENDICATIONS
1 . - Composition comprenant de la streptozotocin ou un de ses dérivés et au moins un inhibiteur de la voie PI3K/AKT/mTOR, et leurs sels, formes ou dérivés pharmaceutiquement acceptables pour la prévention ou le traitement de tumeurs neuro- endrocrines. 1. A composition comprising streptozotocin or a derivative thereof and at least one inhibitor of the PI3K / AKT / mTOR pathway, and pharmaceutically acceptable salts, forms or derivatives thereof for the prevention or treatment of neuroendocrine tumors.
2. - Composition selon la revendication 1 , dans laquelle l'inhibiteur de la voie PI3K/AKT/mTOR est choisi parmi les composés capables d'inhiber l'un des complexes mTORCI et/ou mTORC2 et/ou l'une des enzymes PI3K et/ou Akt. 2. - Composition according to claim 1, in which the inhibitor of the PI3K / AKT / mTOR pathway is chosen from compounds capable of inhibiting one of the mTORCI and / or mTORC2 complexes and / or one of the PI3K enzymes. and / or Akt.
3. - Composition selon la revendication 1 , dans laquelle l'inhibiteur de la voie PI3K/AKT/mTOR est choisi parmi : 3. - The composition according to claim 1, wherein the inhibitor of the PI3K / AKT / mTOR pathway is chosen from:
les inhibiteurs de mTORCI ;  mTORCI inhibitors;
- les inhibiteurs de mTORC2 ;  mTORC2 inhibitors;
les inhibiteurs de PI3K ;  PI3K inhibitors;
les inhibiteurs de Akt ; ou  inhibitors of Akt; or
- les inhibiteurs de PI3K, mTORCI et mTORC2.  inhibitors of PI3K, mTORCI and mTORC2.
4.- Composition selon l'une quelconque des revendications 1 à 3, dans laquelle l'inhibiteur de la voie PI3K/AKT/mTOR est choisi parmi 4. Composition according to any one of claims 1 to 3, in which the inhibitor of the PI3K / AKT / mTOR pathway is chosen from
BEZ235, BKM120, Everolimus (ou RAD001 ), MK-2206 dichlorate, Pictilisib, LY294002, CAL-101 , PI-3065, HS-173, PI-103, NU7441 , TGX-221 , IC-871 14, Wortmannin, XL147, ZSTK474, BYL719, AS-605240, PIK-75, 3-methyladenine, A66, SAR245409, PIK-93, GSK2126458, PIK-90, PF-04691502, AZD6482, Apitolisib, GSK1059615, Duvelisib, Gedatolisib, TG100-1 15, AS-252424, BGT226, CUDC-907, PIK- 294, AS-604850, GSK2636771 , BAY80-6946, YM201636, CH5132799, CAY10505, PIK- 293, PKI-402, TG100713, VS-5584, CZC24832, Quercetin, Rapamycin, AZD8055, Temsirolimus, KU-0063794, Zotarolimus, Torkinib, Tacrolimus, Ridaforolimus, INK 128, IPI-145, SAR245409, Torinl , USI-027, WIE-354, AZD2014, Torin 2, WYE-125132, BET226, Palomid 529, PP121 , PP242 WYE-687, CH5132799, WAY-600, Chrysophanic, ETP-46464, GDC-0349, GDC-0941 , GDC-0068 XL388, Perifosine, GSK690693, Ipatasertib, AZD5363, AT13148, PF-04691502, AT7867, Triciribine, CCT128930, A- 674563, PHT-427, Miltefosine, Honokiol, TIC10, seuls ou en mélange. BEZ235, BKM120, Everolimus (or RAD001), MK-2206 dichloroate, Pictilisib, LY294002, CAL-101, PI-3065, HS-173, PI-103, NU7441, TGX-221, IC-87114, Wortmannin, XL147, ZSTK474, BYL719, AS-605240, PIK-75, 3-methyladenine, A66, SAR245409, PIK-93, GSK2126458, PIK-90, PF-04691502, AZD6482, Apitolisib, GSK1059615, Duvelisib, Gedatolisib, TG100-115, AS -252424, BGT226, CUDC-907, PIK-294, AS-604850, GSK2636771, BAY80-6946, YM201636, CH5132799, CAY10505, PIK-293, PKI-402, TG100713, VS-5584, CZC24832, Quercetin, Rapamycin, AZD8055 , Temsirolimus, KU-0063794, Zotarolimus, Torkinib, Tacrolimus, Ridaforolimus, INK 128, IPI-145, SAR245409, Torin1, USI-027, WIE-354, AZD2014, Torin 2, WYE-125132, BET226, Palomid 529, PP121, PP242 WYE-687, CH5132799, WAY-600, Chrysophanic, ETP-46464, GDC-0349, GDC-0941, GDC-0068 XL388, Perifosine, GSK690693, Ipatasertib, AZD5363, AT13148, PF-04691502, AT7867, Triciribine, CCT128930, A-674563, PHT-427, Miltefosine, Honokiol, TIC10, alone or in admixture.
5. - Composition selon l'une quelconque des revendications 1 à 3, dans laquelle l'inhibiteur de la voie PI3K/AKT/mTOR est choisi parmi BEZ235, BKM120, Everolimus (ou RAD001 ), de préférence Everolimus (ou RAD001 ), BKM120 ou BEZ235, seuls ou en mélange. 5. - Composition according to any one of claims 1 to 3, wherein the inhibitor PI3K / AKT / mTOR pathway is selected from BEZ235, BKM120, Everolimus (or RAD001), preferably Everolimus (or RAD001), BKM120 or BEZ235, alone or in mixture.
6. - Composition selon l'une quelconque des revendications 1 à 5, dans laquelle le ratio en poids de streptozotocin par rapport à l'inhibiteur de la voie PI3K/AKT/mTOR est compris entre 100 et 500 de préférence entre 100 et 300. 6. - Composition according to any one of claims 1 to 5, wherein the weight ratio of streptozotocin relative to the inhibitor of PI3K / AKT / mTOR pathway is between 100 and 500, preferably between 100 and 300.
7.- Composition selon l'une quelconque des revendications 1 à 6, comprenant en outre au moins un excipient pharmaceutiquement acceptable. 7. Composition according to any one of claims 1 to 6, further comprising at least one pharmaceutically acceptable excipient.
8. - Composition selon l'une quelconque des revendications 1 à 7 pour la prévention ou le traitement des tumeurs neuroendocrines du pancréas. 8. - Composition according to any one of claims 1 to 7 for the prevention or treatment of neuroendocrine tumors of the pancreas.
9. - Kit comprenant de la streptozotocin ou un de ses dérivés et au moins un inhibiteur de la voie PI3K/AKT/mTOR, et leurs sels, formes ou dérivés pharmaceutiquement acceptables tels que définis aux revendications 1 à 5, pour son utilisation pour la prévention ou le traitement des tumeurs neuro-endocrines. 9. A kit comprising streptozotocin or a derivative thereof and at least one inhibitor of the PI3K / AKT / mTOR pathway, and their pharmaceutically acceptable salts, forms or derivatives as defined in claims 1 to 5, for its use for the prevention or treatment of neuroendocrine tumors.
10. - Kit selon la revendication 9 pour la prévention ou le traitement de tumeurs neuro-endocrines du pancréas. 10. Kit according to claim 9 for the prevention or treatment of neuroendocrine tumors of the pancreas.
1 1 . - Composition comprenant de la streptozotocin ou un de ses dérivés et au moins un inhibiteur de la voie PI3K/AKT/mTOR, et leurs sels, formes ou dérivés pharmaceutiquement acceptables, dans laquelle l'inhibiteur de la voie PI3K/AKT/mTOR est choisi parmi 1 1. A composition comprising streptozotocin or a derivative thereof and at least one inhibitor of the PI3K / AKT / mTOR pathway, and pharmaceutically acceptable salts, forms or derivatives thereof, wherein the inhibitor of the PI3K / AKT / mTOR pathway is selected among
BEZ235, BKM120, Everolimus (ou RAD001 ), MK-2206 dichlorate, Pictilisib, LY294002, CAL-101 , PI-3065, HS-173, PI-103, NU7441 , TGX-221 , IC-871 14, Wortmannin, XL147, ZSTK474, BYL719, AS-605240, PIK-75, 3-methyladenine, A66, SAR245409, PIK-93, GSK2126458, PIK-90, PF-04691502, AZD6482, Apitolisib, GSK1059615, Duvelisib, Gedatolisib, TG100-1 15, AS-252424, BGT226, CUDC-907, PIK- 294, AS-604850, GSK2636771 , BAY80-6946, YM201636, CH5132799, CAY10505, PIK- 293, PKI-402, TG100713, VS-5584, CZC24832, Quercetin, Rapamycin, AZD8055, Temsirolimus, KU-0063794, Zotarolimus, Torkinib, Tacrolimus, Ridaforolimus, INK 128, IPI-145, SAR245409, Torinl , USI-027, WIE-354, AZD2014, Torin 2, WYE-125132, BET226, Palomid 529, PP121 , PP242 WYE-687, CH5132799, WAY-600, Chrysophanic, ETP-46464, GDC-0349, GDC-0941 , GDC-0068 XL388, Perifosine, GSK690693, Ipatasertib, AZD5363, AT13148, PF-04691502, AT7867, Triciribine, CCT128930, A- 674563, PHT-427, Miltefosine, Honokiol, TIC10, seuls ou en mélange. BEZ235, BKM120, Everolimus (or RAD001), MK-2206 dichloroate, Pictilisib, LY294002, CAL-101, PI-3065, HS-173, PI-103, NU7441, TGX-221, IC-87114, Wortmannin, XL147, ZSTK474, BYL719, AS-605240, PIK-75, 3-methyladenine, A66, SAR245409, PIK-93, GSK2126458, PIK-90, PF-04691502, AZD6482, Apitolisib, GSK1059615, Duvelisib, Gedatolisib, TG100-115, AS -252424, BGT226, CUDC-907, PIK-294, AS-604850, GSK2636771, BAY80-6946, YM201636, CH5132799, CAY10505, PIK-293, PKI-402, TG100713, VS-5584, CZC24832, Quercetin, Rapamycin, AZD8055 , Temsirolimus, KU-0063794, Zotarolimus, Torkinib, Tacrolimus, Ridaforolimus, INK 128, IPI-145, SAR245409, Torinl, USI-027, WIE-354, AZD2014, Torin 2, WYE-125132, BET226, Palomid 529, PP121, PP242 WYE-687, CH5132799, WAY-600, Chrysophanic, ETP-46464, GDC-0349, GDC-0941, GDC-0068 XL388, Perifosine, GSK690693, Ipatasertib, AZD5363, AT13148, PF-04691502 , AT7867, Triciribine, CCT128930, A-674563, PHT-427, Miltefosin, Honokiol, TIC10, alone or in admixture.
12.- Composition selon la revendication 1 1 , dans laquelle, l'inhibiteur de la voie PI3K/AKT/mTOR est choisi parmi BEZ235, BKM120, Everolimus (ou RAD001 ), de préférence everolimus (RAD001 ), BKM120 ou BEZ235, seuls ou en mélange. 12. The composition according to claim 11, wherein the inhibitor of the PI3K / AKT / mTOR pathway is chosen from BEZ235, BKM120, Everolimus (or RAD001), preferably everolimus (RAD001), BKM120 or BEZ235, alone or in mixture.
13.- Composition selon l'une quelconque des revendications 1 1 et 12, dans laquelle le ratio en poids de streptozotocin par rapport à l'inhibiteur de la voie PI3K/AKT/mTOR est compris entre 100 et 500 de préférence entre 100 et 300. 13. A composition according to any one of claims 1 1 and 12, wherein the weight ratio of streptozotocin relative to the PI3K / AKT / mTOR pathway inhibitor is between 100 and 500, preferably between 100 and 300 .
14.- Composition selon l'une quelconque des revendications 1 1 à 13, comprenant en outre au moins un excipient pharmaceutiquement acceptable. 14. A composition according to any one of claims 1 1 to 13, further comprising at least one pharmaceutically acceptable excipient.
15.- Composition selon l'une des revendications 1 à 8 et 1 1 à 14 ou kit selon les revendications 9 ou 10 pour une administration simultanée, séparée ou séquentielle à un patient qui en a besoin, notamment pour la prévention ou le traitement de tumeurs neuroendocrines, de préférence de tumeurs neuroendocrines du pancréas. 15.- composition according to one of claims 1 to 8 and 1 1 to 14 or kit according to claims 9 or 10 for simultaneous, separate or sequential administration to a patient who needs, especially for the prevention or treatment of neuroendocrine tumors, preferably neuroendocrine tumors of the pancreas.
PCT/EP2016/055253 2015-03-11 2016-03-11 Composition for treating pancreatic neuroendocrine tumours WO2016142508A1 (en)

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