ASSOCIATION BETWEEN 3-[(3-{[4-(4-MORPHOLINYLMETHYL)-1H-PYRROL-2-YL]METHYLENE}-2-OXO-2,3-DIHYDRO-1H-INDOL-5-YL)METHYL]-1,3-THIAZOLIDINE-2,4-DIONE AND AN EGFR TYR KINASE INHIBITOR

Abstract

Association between 3-[(3-{[4-(4-morpholinylmethyl)-1 h -pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1 h -indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione of formula (i): or one of its z or e isomers and / or addition salts to a pharmaceutically acceptable acid or base, and a human epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, and its use in the treatment of non-small cell lung cancer.

Description

[0001] The present invention relates to the association between 3-[(3-{[4-(4-morpholinylmethyl)-1 H -pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1 H -indol-5-yl)methyl] -1,3-thiazolidine-2,4-dione of formula (I): or one of its Z or E isomers and / or addition salts to a pharmaceutically acceptable acid or base, and a human epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, as well as its use for the treatment of non-small cell lung cancer, and more particularly in patients resistant to an EGFR tyrosine kinase inhibitor. Non-small cell lung cancer is now the leading cause of cancer death worldwide (Goldstraw, P., D. Ball, JR Jett, CT Le, E. Lim, AG Nicholson, and FA Shepherd, 2011, Non-small-cell lung cancer: Lancet, v. 378, no. 9804, p. 1727-1740; Jemal, A., F. Bray, MM Center, J. Ferlay, E. Ward, and D. Forman, 2011, Global cancer statistics: CA Cancer J Clin, v. 61, no. 2, p. 69-90). At the time of diagnosis, the majority of patients have advanced disease, with a one-year survival rate of 30% and a five-year survival rate of 10% (USNational Institutes of Health, National Cancer Institute (http: / / seer.cancer.gov / archive / csr / 1975_2011 / results_merged / topic_delaygraphs_overview.pdf; http: / / www.cancerresearchuk.org / cancer-info / cancerstats / types / lung / survival / lung-cancer-survival-statistics). Activating mutations in the EGFR gene lead to oncogenic addiction, meaning the cancer cell's dependence on this abnormality for its growth and survival. These mutations are common in lung adenocarcinoma, occurring in 15% of cases in Caucasian patients and 40-50% of cases in Asian patients (Shigematsu, H. et al., 2005, Clinical and biological features associated with epidermal growth factor receptor gene mutations in lung cancers: J Natl Cancer Inst, v. 97, no. 5, p. 339-346).In patients with EGFR gene mutations, EGFR tyrosine kinase inhibitors significantly delay disease progression compared to chemotherapy and are considered the standard of care. Current treatments on the market include gefitinib and erlotinib for first-generation inhibitors, and afatinib for second-generation inhibitors, each generation targeting active EGFR mutations. Unfortunately, most patients relapse after several months of treatment due to the acquisition of other genetic and protein alterations capable of generating resistance to EGFR tyrosine kinase inhibitors. Several resistance mechanisms have been identified, and in particular, a novel mutation of the EGFR receptor (T790M: substitution of threonine by methionine at position 790) is predominantly found in patients resistant to treatment.After resistance to EGFR tyrosine kinase inhibitors develops, the prognosis becomes very poor, and patients are offered chemotherapy with a low efficacy rate. In this context, the search for new therapeutic alternatives in non-small cell lung cancer, and particularly in patients resistant to EGFR tyrosine kinase inhibitors, with the aim of improving progression-free survival, remains a pressing issue. In particular, re-sensitizing patients resistant to EGFR tyrosine kinase inhibitors represents a promising therapeutic strategy to explore. Third-generation EGFR tyrosine kinase inhibitors are currently being developed to act specifically in patients who have acquired a secondary mutation, such as T790M, and appear to restore the activity of the treatment.Other alternatives, acting on resistance pathways other than the T790M mutation and / or involving other cellular receptors, are still needed and highly anticipated by patients, and may include combinations with third-generation inhibitors. X. Wu et al. reported in Oncotarget, 5(20), 9546, 2014, that cancer sensitivity after resistance can be restored by targeting AXL. SGI17079, an AXL inhibitor, in combination with erlotinib, reverses erlotinib resistance in non-small cell lung cancer. Rho et al. They report in Cancer Research, 74(1), 253, 2013, that the combination of Gefitinib or Erlotinib with NPS-1034 inhibits cell proliferation and induces cell death in non-small cell lung cancer cells resistant to Gefitinib or Erlotinib in a synergistic manner. NPS-1034 targets both MET and AXL.WO-A-2014 / 138364 discloses a combination of an FGFR inhibitor and an EGFR tyrosine kinase inhibitor for the treatment of non-small cell lung cancer in patients with EGFR mutations. In all that follows, and where nothing else is specified, by "3-[(3-{[4-(4-morpholinyl-methyl)-1. H -pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1 H -indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione" we hear "3-[(3- {[4-(4-morpholinyl-methyl)-1 H -pyrrol-2-yl] methylene}-2-oxo-2,3-dihydro-1 H -indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione, as well as its Z or E isomers and / or salts of addition to a pharmaceutically acceptable acid or base.

[0002] The 3-[(3-{[4-(4-morpholinylmethyl)-1 H -pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1 H-indol-5-ylmethyl-1,3-thiazolidine-2,4-dione is a potent inhibitor of cancer cell migration, particularly useful for the treatment of cancers, especially metastatic solid tumors. It is described in patent applications WO2011 / 015728 and WO2015 / 004395.

[0003] According to the invention, it has been shown that the effects of 3-{[(3-{[4-(4-morpholinylmethyl)-1 H- pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1 H -indol-5-yl]methyl}-1,3-thiazolidine-2,4-dione enabled the overcoming of resistance to EGFR tyrosine kinase inhibitors in animal models previously treated with such an inhibitor.

[0004] These effects suggest the possibility of using the combination of 3-{[(3-{[4-(4-morpholinylmethyl)-1 H -pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1 H-indol-5-yl]methyl} -1,3-thiazolidine-2,4-dione, and an EGFR tyrosine kinase inhibitor in the treatment of non-small cell lung cancer, particularly in patients in whom disease progression or relapse has been observed despite treatment.

[0005] More specifically in the association according to the invention, the 3-[(3-{[4-(4-morpholinyl-methyl)-1 H -pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1 H -indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione is in the form of the Z isomer.

[0006] Preferably in the association according to the invention, the 3-[(3- {[4-(4-morpholinyl-methyl)-1 H -pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1 H -indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione is in the form of a salt, and in particular a hydrochloride or a mesylate.

[0007] Even more advantageously, the association according to the invention contains 3-[((3Z)-3-{[4-(4-morpholinyl-methyl)-1H -pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1 H -indol-5-yl) methyl]-1,3-thiazolidine-2,4-dione mesylate.

[0008] Among the EGFR tyrosine kinase inhibitors according to the invention, we can mention Erlotinib, Gefitinib and Afatinib for first and second generation inhibitors, and AZD9291 (Osimertinib) or Rociletinib for third generation inhibitors.

[0009] According to an advantageous embodiment, the EGFR tyrosine kinase inhibitor of the combination according to the invention is N-(3-ethynylphenyl)-6,7-di(2-methoxyethoxy)quinazolin-4-amine or Erlotinib of formula (II): or one of its salts of addition to a pharmaceutically acceptable acid or base, and in particular its hydrochloride.

[0010] According to another advantageous embodiment, the EGFR tyrosine kinase inhibitor of the association according to the invention is the N-(3-chloro-4-fluoro-phenyl)-7-methoxy-6-(3-morpholin-4-ylpropoxy)quinazolin-4-amine or Gefitinib of formula (III): or one of its salts of addition to a pharmaceutically acceptable acid or base. Preferably, the association according to the invention relates to the Z isomer of 3-[(3-{[4-(4-morpholinyl-methyl1)-1 H -pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1 H -indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione or one of its pharmaceutically acceptable salts, with Gefitinib or one of its pharmaceutically acceptable salts.

[0011] More specifically, the invention relates to the association between 3-[((3Z)-3-{[4-(4-morpholinyl-methyl)-1 H - pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1 H -indol-5-yl) methyl]-1,3-thiazolidine-2,4-dione mesylate, with Gefitinib or one of its pharmaceutically acceptable salts.

[0012] The invention also relates to pharmaceutical compositions containing the combination of 3-[(3- {[4-(4-morpholinyl-methyl)-1 H -pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1 H -indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione, and an EGFR tyrosine kinase inhibitor in combination with one or more pharmaceutically acceptable excipients.

[0013] The invention also relates to said pharmaceutical compositions for their use in the treatment of non-small cell lung cancer, and more particularly in patients resistant to an EGFR tyrosine kinase inhibitor.

[0014] Among the pharmaceutical compositions according to the invention, particular attention will be paid to those suitable for administration by oral, parenteral, intramuscular and intravenous routes, percutaneous or transcutaneous, nasal, rectal, sublingual, ocular, respiratory routes and more specifically plain or coated tablets, sublingual tablets, capsules, lozenges, caplets, tablets, injectable preparations, aerosols, eye or nasal drops, suppositories, creams, ointments, dermal gels, etc.

[0015] In a preferred embodiment, the 3-[(3-{[4-(4-morpholinyl-methyl)-1 H -pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1 H -indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione is administered orally.

[0016] Besides 3-[(3- {[4-(4-morpholinyl-methyl)-1 H -pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1 H-indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione and the EGFR tyrosine kinase inhibitor, the pharmaceutical compositions according to the invention contain one or more excipients or vehicles selected from diluents, lubricants, binders, disintegrating agents, stabilizers, preservatives, absorbents, colorants, sweeteners, flavorings, etc...

[0017] For example, we can cite: for diluents: lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycerin; for lubricants: silica, talc, stearic acid and its magnesium and calcium salts, polyethylene glycol; for binders: aluminum magnesium silicate, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidone; for disintegrants: agar, alginic acid and its sodium salt, effervescent mixtures.

[0018] The compounds in the combination can be administered simultaneously or sequentially. The corresponding pharmaceutical compositions may allow for the immediate or delayed release of the active ingredients. Furthermore, the compounds in the combination may be administered as two separate pharmaceutical compositions, each containing one of the active ingredients, or as a single pharmaceutical composition in which the active ingredients are mixed.

[0019] The effective dosage varies depending on the patient's sex, age, and weight, the route of administration, the type of cancer, and any concomitant treatments, and ranges from 300 to 1500 mg base equivalents of 3-[(3-{[4-(4-morpholinyl-methyl)-1H-pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1H-indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione per day, and more preferably between 400 and 800 mg base equivalents per day, and even more specifically between 500 and 600 mg base equivalents per day. The dose of the EGFR tyrosine kinase inhibitor will be equal to or lower than that used when administered alone. For example, in the case of Gefitinib, the administered dose is 250 mg per day. For Erlotinib, it is 25 to 150 mg per day. PHARMACEUTICAL COMPOSITION 1000 tablets dosed at 100 mg equivalent base of 3-[3-{[4-(4-morpholinylmethyl)-1 H -pyrrol-2-yl] methylene}-2-oxo-2,3-dihydro-1 H -indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione methanesulfonate, Z isomer 121 g Sodium starch glycolate 20 g Corn starch 133 g Lactose monohydrate 357 g Magnesium stearate 6,7 g Silica 1,3 g Povidone 46,6 g PRECLINICAL STUDIES A) Cell viability assay on the erlotinib-resistant HCC827 cell line

[0020] A cell viability assay was used to measure the antiproliferative activity of antitumor compounds. The cell line chosen was HCC827, a non-small cell lung cancer cell line dependent on EGFR for survival. The parameter used was the IC50, which is the concentration of the product inhibiting 50% of cell proliferation compared to untreated control cells. Cells were seeded (150 µl) at the appropriate density two days before the experiment in the wells of 96-well plates. One column contained untreated control cells representing 100% proliferation. The others were incubated with the products to be tested for four doubling times. The median inhibitory concentration of the EGFR tyrosine kinase inhibitor erlotinib for cell viability of the HCC827 cell line was 10 nM.Acquired resistance to erlotinib is generated by chronic exposure of the HCC827 cell line to erlotinib: cells are exposed to erlotinib at a dose of 1 µM in culture medium until the doubling time stabilizes, approximately 2 months. The median inhibitory concentration of erlotinib for cell viability of the resistant HCC827 cell line is then approximately 1000 times higher at 11.5 µM. The resistant cells are then exposed to 3-[(3-{[4-(4-morpholinyl-methyl)-1H-pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1H-indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione hydrochloride at a concentration of 100 nM in combination with increasing doses of erlotinib. 3-[(3- {[4-(4-morpholinyl-methyl)-1H-pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1H-indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione hydrochloride, alone, has no effect on viability.In combination, the median inhibitory concentration of Erlotinib on the resistant HCC827 cell line returns to the order of that of the non-resistant HCC827 cell line at 3.8 nM.

[0021] This result shows that 3-[(3-{[4-(4-morpholinyl-methyl)-1H-pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1H-indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione is able to restore sensitivity to an EGFR tyrosine kinase inhibitor in a non-small cell lung cancer cell line resistant to such an inhibitor. B) Inhibition of the growth of the HCC827 tumor resistant to Erlotinib

[0022] The HCC827 non-small cell lung cancer cell line, made resistant in vitroErlotinib was grafted subcutaneously into female SCID mice at a rate of 5 x 10⁶ cells per mouse. Tumors were randomized into groups of eight mice when the tumor volume reached approximately 200 mm³. Daily treatments with compound A (3-[(3-{[4-(4-morpholinyl-methyl)-1H-pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1H-indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione hydrochloride) at a dose of 50 mg / kg, and with erlotinib at a dose of 12.5 mg / kg, were administered orally (vehicles = ammonium acetate / HEC and PEG300 / ethanol / water buffers, respectively) over a period of 19 days as indicated by the triangles on the figure 1 Tumor volumes were measured two to three times per week using calipers. Median tumor volumes with interquartile ranges are shown on the graph.

[0023] At the end of treatment, on day 19, growth inhibition after treatment with the compounds alone was 65% for erlotinib and 83% for 3-[(3-{[4-(4-morpholinyl-methyl)-1H-pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1H-indo1-5-yl)methyl]-1,3-thiazolidine-2,4-dione hydrochloride (Compound A). When the two compounds were used in combination, tumors regressed completely, and this regression persisted over time, after treatment cessation until day 30. The synergy observed between the two products was statistically significant over the study period (p<0.001).

[0024] The results are reported in the Figure 1 . ETUDE CLINICAL

[0025] Patients with non-small cell lung cancer develop resistance to treatment with an EGFR tyrosine kinase inhibitor (gefitinib, erlotinib, afatinib, osimertinib, or rociletinib), becoming unresponsive to treatment and causing disease progression. A proof-of-concept study is underway to confirm the results observed in preclinical studies and demonstrate that 3-[(3-{[4-(4-morpholinyl-methyl)-1H-pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1H-indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione can restore sensitivity to an EGFR inhibitor, specifically gefitinib, in patients with non-small cell lung cancer who have become resistant. Patients are included based on their molecular profile. This study includes a phase I component aimed at evaluating the safety profile and determining the recommended dose for further development. Approximately 20 patients will be included.During this phase, patients will be treated in 28-day cycles with a dose of 400, 500, or 600 mg per day of the base equivalent of 3-[(3-{[4-(4-morpholinyl-methyl)-1H-pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1H-indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione in combination with 250 mg per day of gefitinib. Treatment will be continued until disease progression. Following this phase, a phase II trial will be initiated to evaluate the activity of the combination of 3-[(3-{[4-(4-morpholinyl-methyl)-1H-pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1H-indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione and gefitinib. This phase II trial will include approximately 150 patients. Tumor assessment will be performed every two months.Patients will be treated in 28-day cycles with the recommended dose of 3-[(3-{[4-(4-morpholinyl-methyl)-1H-pyrrol-2-yl]methylene} -2-oxo-2,3-dihydro-1H-indol-5-yl) methyl]-1,3-thiazolidine-2,4-dione defined in Phase I in combination with 250 mg of Gefitinib per day.

Claims

1. Association between 3-[(3-{[4-(4-morpholinylmethyl)-1H-pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1H-indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione of formula (I): and its Z or E isomers and / or addition salts with a pharmaceutically acceptable acid or base, and a human epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor.

2. Association according to claim 1, characterised in that 3-[(3-{[4-(4-morpholinylmethyl)-1H-pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1H-indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione is used in the form of the Z isomer.

3. Association according to either claim 1 or claim 2, characterised in that 3-[(3-{[4-(4-morpholinylmethyl)-1H-pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1H-indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione is used in the form of a hydrochloride.

4. Association according to either claim 1 or claim 2, characterised in that 3-[(3-{[4-(4-morpholinylmethyl)-1H-pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1H-indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione is used in the form of a mesylate.

5. Association according to any one of claims 1 to 4, characterised in that the EGFR tyrosine kinase inhibitor is gefitinib or erlotinib.

6. Association according to any one of claims 1 to 5 for use in the treatment of non-small-cell lung cancer.

7. Association according to any one of claims 1 to 5 for use in the treatment of non-small-cell lung cancer in patients who are resistant to an EGFR tyrosine kinase inhibitor.

8. Pharmaceutical composition comprising as active ingredient 3-[(3-{[4-(4-morpholinylmethyl)-1H-pyrrol-2-yl]methylene}-2-oxo-2,3-dihydro-1H-indol-5-yl)methyl]-1,3-thiazolidine-2,4-dione or a Z or E isomer thereof and / or an addition salt thereof with a pharmaceutically acceptable acid or base, in association with an EGFR tyrosine kinase inhibitor according to any one of claims 1 to 7 in combination with one or more pharmaceutically acceptable excipients.

9. Pharmaceutical composition according to claim 8 for use in the treatment of non-small-cell lung cancer.

10. Pharmaceutical composition according to claim 9 for use in the treatment of non-small-cell lung cancer in patients who are resistant to an EGFR tyrosine kinase inhibitor.