Combination of RAS inhibitors and farnesyltransferase inhibitors for the treatment of cancer - Patent Application 20070233633

Abstract

KRAS G12C mutations can be found in approximately 13% of patients with non-small cell lung cancer (NSCLC) and have historically been associated with poor prognosis. Currently, data from Phase II trials show efficacy of the new KRAS G12C-specific inhibitor sotorasib in patients with advanced stage NSCLC. However, it can be expected that resistance to Ras inhibitors may occur and there is a demand to identify new therapeutic avenues to counteract said resistance. The inventors herein show that sotorasib and tipifarnib, when used alone, did not show significant antitumor effects against lung cancer cells with G12C KRAS mutations, but the combination potently induced cell death, suggesting synergy between these agents. Thus, the present invention relates to a combination of a Ras inhibitor and a farnesyltransferase inhibitor for the treatment of cancer.

Description

[Technical field]

[0001] The present invention is in the field of medicine, particularly oncology. [Background technology]

[0002] Lung cancer is the leading cause of cancer deaths worldwide. Metastatic non-small cell lung cancer (NSCLC) has benefited in recent years from two successive breakthroughs: the identification of oncogenic drivers such as KRAS mutations leading to the development of targeted therapies, and the understanding of the cancer-immunity cycle leading to the development of immune checkpoint inhibitors. KRAS G12C mutations can be found in approximately 13% of patients with non-small cell lung cancer (NSCLC) and have historically been associated with poor prognosis. Currently, data from a phase II study show efficacy of the novel KRAS G12C-specific inhibitor sotorasib in patients with advanced stage NSCLC harboring this alteration who have progressed on at least one standard of care treatment. However, it can be expected that resistance to Ras inhibitors may occur as with other targeted therapies (e.g., EGFR), and that there is a demand to identify new therapeutic avenues to counteract the above-mentioned resistance. Recently, farnesyltransferase inhibition has been shown to overcome the adaptive resistance to osimertinib in EGFR-mutant NSCLC (Sarah Figarol, Celia Delahaye, Remi Gence, Raghda Asslan, Sandra Pagano, Claudine Tardy, Jacques Colinge, Jean-Philippe Villemin, Antonio Maraver, Irene Ferrer, Luis Paz-Ares, Isabelle Lajoie-Mazenc, Estelle Clermont, Anne Casanova, Anne Pradines, Julien Mazieres, Olivier Calvayrac, Gilles Favre; Farnesyltransferase inhibition overcomes the adaptive resistance to osimertinib in EGFR-mutant NSCLC; bioRxiv 2022.04.01.486707;doi:https: / / doi.org / 10.1101 / 2022.04.01.486707). Summary of the Invention

[0003] The present invention is defined by the claims. In particular, the present invention relates to the combination of a Ras inhibitor and a farnesyltransferase inhibitor for the treatment of cancer. [Brief description of the drawings]

[0004] [Figure 1] Figure 1 shows that sotorasib and tipifarnib synergize in cell death induction. A. Cell culture density was measured twice weekly for 35 days of treatment as indicated. B. Amido black staining of 6-well plates after 35 days of treatment as indicated. [Diagram 2] Figure 2 shows that tipifarnib abrogates the development of resistant proliferative clones (RPCs) induced by sotorasib. H23 (A-B) and Calu-1 (C-D) KRAS (G12C) mutant non-small cell lung cancer (NSCLC) cell lines were transduced with the FUCCI (Fluorescent Ubiquitination-Based Cell Cycle Indicator) system and response / relapse to sotorasib (1 μM) or sotorasib (1 μM) + tipifarnib (1 μM) was monitored for 50 days by Incucyte® measuring total cell numbers (A and C) or cell cycle kinetics (B and D). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0005] Key Definitions As used herein, the term "Ras" has its general meaning in the art and refers to any member of the Ras family of proteins, or mutants thereof. Ras family proteins include, but are not limited to, HRAS, KRAS, and NRAS, as well as other members of this subfamily, DIRAS1; DIRAS2; DIRAS3; ERAS; GEM; MRAS; NKIRAS1; NKIRAS2; NRAS; RALA; RALB; RAP1A; RAP1B; RAP2A; RAP2B; RAP2C; RASD1; RASD2; RASL10A; RASL10B; RASL11A; RASL11B; RASL12; REM1; REM2; RERG; RERGL; RRAD; RRAS; RRAS2 (Wennerberg et al., The Ras superfamily at a glance, J. Cell. Sci., 2005, 118 (Pt 5), 843-846).

[0006] As used herein, the term "mutant Ras cancer" refers to a cancer in which the cancer cells contain an activating mutation in the Ras protein.

[0007] As used herein, the term "Ras mutation" refers to an activating mutation in the ras gene or Ras protein. A Ras mutation can refer to either a genetic change in the DNA sequence of one of the ras genes that results in activation of the corresponding Ras protein, or a change in the amino acid sequence of the Ras protein that results in activation. In particular, the Ras mutation is a KRAS mutation. As used herein, the term "KRAS mutation" includes any one or more mutations in the KRAS (which may also be called KRAS2 or RASK2) gene. For example, the KRAS mutation is located in exon 3 or exon 4 of the gene. Examples of KRAS mutations include, but are not limited to, G12C, G12D, G13D, G12R, G12S, and G12V. KRAS is one of the oncogenes that frequently mutate in human cancers. In particular, KRAS mutations are found in 30-40% of tumors and, together with APC, represent one of the somatic alterations involved in the initiation of colorectal cancer. This mutation occurs early in the carcinogenesis process and is maintained at various stages of disease progression, such as lymph node involvement and metastatic spread. Recent studies involving a large number of patients have shown that mutated KRAS is associated with poor prognosis in colorectal cancer progression, with the effect being more evident in stage II and III disease (Nash, et al, Ann. Surg. Oncol, 17:416-424, 2010). In the same group, other studies (Nash, et al, Ann. Surg. Oncol, 17:572-578, 2010) have shown that KRAS mutations are associated with more rapid and aggressive metastatic behavior in liver metastases from the colorectum. Furthermore, KRAS mutations have been reported to result in drug resistance and treatment failure to epidermal growth factor receptor (EGFR)-targeted therapeutics in metastatic colorectal cancer.KRAS mutations confer resistance to both cetuximab (Erbitux®) and panitumumab (Vectibix®) (Allegra et al, J. Clin. Oncol, 27:2091-2096, 2008; Linardou et al, Lancet Oncol, 9:962-972, 2008). Numerous mutations in NRAS are known, typically including Q61R, Q61K, Q61H, Q61L, Q61N, Q61E, Q61P, A146T, A146P, or A146V.

[0008] As used herein, the term "Ras inhibitor" refers to any compound that (i) directly interacts with RAS, for example by binding to RAS, and any compound that reduces the expression or activity of RAS. In some embodiments, the Ras inhibitor is not tipifarnib. In particular, it refers to an inhibitor of Ras kinase membrane translocation and activity. Ras inhibitors can be any type of molecule, including but not limited to small molecules, antibodies, and expression regulators (e.g., antisense molecules, microRNAs, siRNAs, aptamers, etc.), which can directly act on Ras protein, interfere with the expression (e.g., transcription, splicing, translation, and / or post-translational processing) of Ras protein, and / or abrogate the inappropriate subcellular localization and / or membrane translocation and / or phosphorylation and / or activation of Ras protein. Methods for determining whether a compound is a Ras inhibitor are well known (e.g., Haider K, Sharma A, Yar MS, Yakkala PA, Shafi S, Kamal A. Novel approaches for the development of direct KRas inhibitors: structural insights and drug design. Expert Opin Drug Discov. 2022 Mar;17(3):247-257. doi:10.1080 / 17460441.2022.2029842. Epub 2022 Jan 27. PMID:35084268).

[0009] As used herein, the term "resistant to Ras inhibitor" is used in a very broad context to refer to the reduced effectiveness of at least one Ras inhibitor that inhibits cell growth, kills cells, or inhibits one or more cell functions, and the ability of a cell to survive exposure to a substance designed to inhibit cell growth, kill cells, or inhibit one or more cell functions. The resistance exhibited by a cell can be acquired, for example, by previous exposure to the substance, or can be inherent or natural. The resistance exhibited by a cell can be complete, in that the substance becomes completely ineffective against the cell, or partial, in that the substance reduces its effectiveness. Thus, the term "resistant" refers to the frequent onset of cancer, or the progression of cancer, regardless of whether the disease has been cured before onset or progression.

[0010] As used herein, the terms "persistent cells," "cancer persister cells," "drug-resistant persisters," and "DTPs" are used to refer to a small population of cancer cells that maintain viability upon anti-cancer targeted therapeutic treatment, particularly treatment with Ras inhibitors. More specifically, this refers to cancer cells that are resistant to high concentrations of Ras inhibitor treatment when used at concentrations 100-fold higher than the IC50. These cells are low-proliferative and largely quiescent.

[0011] As used herein, the term "drug-resistant proliferator-retaining survivors," or "drug-resistant cells," as used herein, refers to cancer cells that are capable of proliferating upon continued high concentrations of cancer drug treatment, particularly treatment with Ras inhibitors.

[0012] As used herein, the term "recurrence" refers to the reappearance of cancer after an initial response (e.g., complete or partial response) period. The initial response period may involve a level of cancer cells falling below a predefined threshold, e.g., below 20%, 15%, 10%, 5%, 4%, 3%, 2%, or 1%. The reappearance may involve a level of cancer cells rising above a predefined threshold, e.g., above 20%, 15%, 10%, 5%, 4%, 3%, 2%, or 1%. More generally, the response (e.g., complete or partial response) may involve no detectable MRD (minimal residual disease). In some embodiments, the initial response period lasts for at least 1, 2, 3, 4, 6, 8, 10, or 12 months, or at least 1, 2, 3, 4, or 5 years.

[0013] As used herein, the term "farnesyltransferase inhibitor" refers to a molecule that inhibits enzyme-catalyzed transfer of a farnesyl residue to a substrate. As used herein, the substrate to be farnesylated is typically a polypeptide that is at least four amino acids in length. The polypeptide to be enzyme-catalyzed to be farnesylated preferably contains a CAAX sequence motif, where C corresponds to a cysteine ​​moiety, A corresponds to an aliphatic amino acid moiety, and X corresponds to another amino acid moiety specified by the enzyme that catalyzes farnesylation. As used herein, enzyme-catalyzed transfer of a farnesyl residue refers to a biochemical reaction in which a farnesyl residue is transferred to a substrate, preferably a polypeptide. The enzyme that catalyzes the transfer of a farnesyl residue to a substrate is called a farnesyltransferase. In this case, typically, activated farnesol is transferred. The activated farnesol is preferably farnesyl diphosphate (farnesyl pyrophosphate, FPP). Typically, the polypeptide corresponding to the substrate is farnesylated at the cysteine ​​moiety. Thus, a thiol ester is generated. The terms "thiol ester" and "thioester" are interchangeable and refer to the group R1-CO-S-R2, where thiol ester can also include the tautomeric form of the ester R1-COH=S-R2. Preferably, the cysteine ​​moiety that can be farnesylated is located close to the C-terminus of the protein. Particularly preferably, the cysteine ​​moiety of the CAAX sequence motif is farnesylated, where C corresponds to the cysteine ​​moiety, A corresponds to the aliphatic amino acid moiety, and X corresponds to another amino acid moiety specified by the enzyme that catalyzes farnesylation. The enzyme that catalyzes farnesylation is preferably a farnesyltransferase (FTase), which corresponds to a prenyltransferase having the enzyme classification number EC2.5.1.X, more preferably EC2.5.1.29, EC2.5.1.58, or EC2.5.1.59, even more preferably EC2.5.1.29 or EC2.5.1.58. The enzyme typically binds one or more zinc ions (Zn2+).Geranylgeranyltransferase may also be effective as farnesyltransferase in the context of the present invention, because this enzyme can also farnesylate certain polypeptides. Any substance or any molecular component that can slow down or inhibit enzyme-catalyzed farnesylation may be a farnesyltransferase inhibitor. Preferably, lowering the farnesylation rate can be understood as slowing down by more than 10%, more preferably more than 25%, even more preferably more than 50%, even more preferably more than 75%, even more preferably more than 80%, even more preferably more than 90%, and most preferably more than 95% by adding an appropriate concentration of farnesyltransferase inhibitor at the action site, compared to a similar reaction environment without adding a farnesyltransferase inhibitor. More importantly, farnesyltransferase inhibitors inhibit the farnesylation of RhoB. As used herein, the term "RhoB" has its general meaning in the art and refers to member B of the ras homolog gene family, a protein encoded by the RHOB gene in humans.

[0014] As used herein, the term "combination" is used to refer to any administration form that provides a first agent together with an additional (second, third, etc.) agent. The agents can be administered together, separately, or sequentially, and in any order. The agents administered in combination have biological activity in the subject to which they are delivered. Thus, within the context of the present invention, a combination comprises at least two different agents, one agent being at least a Ras inhibitor and the other agent being a farnesyltransferase inhibitor. In some cases, the combination of the present invention causes synthetic lethality of cancer cells, particularly DTCs.

[0015] As used herein, the phrase "therapeutically effective amount" refers to an amount effective at the dosage and duration required to obtain the desired therapeutic result. The therapeutically effective amount of an agent may vary depending on factors such as the individual's medical condition, age, sex, and weight, and the ability of the agent to induce the desired response in the individual. A therapeutically effective amount is also one in which any toxic or harmful effects of the antibody or antibody portion are outweighed by the therapeutically beneficial effects. The effective dosage and administration regime of the agent depends on the disease or condition being treated and can be determined by one of ordinary skill in the art. A physician who is skilled in the art can easily determine and prescribe the effective amount of the pharmaceutical composition required. For example, a physician can start the dosage of the agent used in the pharmaceutical composition at a level lower than that required to achieve the desired therapeutic effect, and gradually increase the dosage until the desired effect is achieved. In general, the appropriate dosage of the composition of the present invention may be the amount of the compound that is the lowest dosage effective to produce a therapeutic effect with a particular administration regimen. Such an effective dosage may generally depend on the factors described above. For example, a therapeutically effective amount for therapeutic use may be determined by its ability to stabilize the progression of a disease. A therapeutically effective amount of a therapeutic compound can reduce tumor size or otherwise ameliorate symptoms in a subject. One of skill in the art can determine such amounts based on factors such as the size of the subject, the severity of the subject's symptoms, and the particular composition or route of administration selected. An exemplary, non-limiting range for a therapeutically effective amount of an agent is about 0.1-100 mg / kg, such as about 0.1-50 mg / kg, such as about 0.1-20 mg / kg, such as about 0.1-10 mg / kg, such as about 0.5, such as about 0.3, about 1, about 3 mg / kg, about 5 mg / kg, or about 8 mg / kg. An exemplary, non-limiting range for a therapeutically effective amount of an antibody of the invention is 0.02-100 mg / kg, such as about 0.02-30 mg / kg, such as about 0.05-10 mg / kg, or 0.1-3 mg / kg, such as about 0.5-2 mg / kg. Administration can be, for example, intravenous, intramuscular, intraperitoneal, or subcutaneous, for example, proximal to the target site. The administration regimen in the above-mentioned treatment methods and uses is adjusted to provide the optimum desired response (e.g., a therapeutic response).For example, a single bolus may be administered, several divided doses may be administered over time, or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. In some embodiments, the effect of treatment is monitored during treatment, e.g., at predetermined time points. As a non-limiting example, treatments according to the invention can be administered daily on at least one day on the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th, 27th, 28th, 29th, 30th, 31st, 32nd, 33rd, 34th, 35th, 36th, 37th, 38th, 39th, or 40th day, or alternatively on at least one week on the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, or 20th week, or any combination thereof, after initiation of treatment. A daily dosage of the agent of the invention in an amount of about 0.1 to 100 mg / kg, e.g., 0.2, 0.5, 0.9, 1.0, 1.1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, 45, 50, 60, 70, 80, 90, or 100 mg / kg, can be provided using a single dose, or divided doses every 24, 12, 8, 6, 4, or 2 hours, or any combination thereof.

[0016] The term "kit" or "combination preparation" as used herein specifically defines a "kit of parts" in the sense that the combination partners as defined above can be taken individually or by using various predefined combinations containing specific amounts of the combination partners, i.e., together or at various times. The parts of the kit of parts can then be administered, for example, together or chronologically staggered, i.e., at different times and with the same or different time intervals for any part of the kit of parts. The ratio of the total amounts of the combination partners administered in a combination preparation can vary. The combination partners can be administered by the same or different routes.

[0017] Methods of the Invention A first object of the present invention relates to a method for treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective combination comprising a Ras inhibitor and a farnesyltransferase inhibitor.

[0018] A further object of the present invention relates to a method for delaying and / or preventing the onset of a cancer resistant to a Ras inhibitor in a subject, comprising administering to the subject a therapeutically effective amount of a Ras inhibitor in combination with a farnesyltransferase inhibitor.

[0019] A further object of the present invention relates to a method of treating a cancer that is resistant to a Ras inhibitor in a subject in need of such treatment, comprising administering to the subject a therapeutically effective amount of a farnesyltransferase inhibitor.

[0020] A further object of the present invention relates to a method for abrogating resistance to an administered Ras inhibitor in a subject suffering from cancer, comprising administering to the subject a therapeutically effective amount of a farnesyltransferase inhibitor.

[0021] A further object of the present invention relates to a method for enhancing the efficacy of a Ras inhibitor administered as part of a treatment regimen to a subject suffering from cancer, comprising administering to the subject a pharmacologic effective amount of a farnesyltransferase inhibitor in combination with the Ras inhibitor.

[0022] A further object of the present invention relates to the use of farnesyltransferase inhibitors to inhibit or suppress the proliferation of cancer persister cells or the colony formation of cancer persister cells, thereby preventing or delaying the development of cancer recurrence and / or acquired resistance to treatment with Ras inhibitors. Moreover, this effect on cancer persister cells may allow a complete response to cancer treatment. In fact, farnesyltransferase inhibitors may eliminate cancer persister cells. Also relates to a method of removing or reducing the cancer persister cell population and / or preventing or delaying the development of cancer recurrence and / or acquired resistance to cancer treatment, comprising removing or reducing the cancer persister cell population by administering a therapeutically effective amount of a farnesyltransferase inhibitor. Farnesyltransferase inhibitors may be useful in targeting tumor persister cells, and thus may prevent the development of drug-resistant clones, especially in the context of combination therapy with Ras inhibitors. Thus, the farnesyltransferase inhibitors of the present invention are particularly suitable for eradicating drug-resistant growth persisters.

[0023] In the present invention, the patient suffers from Ras mutant cancer. The scope of the present invention includes carcinomas such as bladder cancer (including advanced and metastatic bladder cancer), breast cancer, colon cancer (including colorectal cancer), kidney cancer, liver cancer, lung cancer (including small cell lung cancer and small cell lung cancer and lung adenocarcinoma), ovarian cancer, prostate cancer, testicular cancer, urogenital cancer, lymphatic system cancer, rectal cancer, laryngeal cancer, pancreatic cancer (including exocrine pancreatic cancer), esophageal cancer, stomach cancer, gallbladder cancer, cervical cancer, thyroid cancer, and skin cancer (including squamous cell carcinoma); hematopoietic tumors of lymphoid lineage, such as leukemia, acute lymphocytic carcinoma, and acute lymphocytic carcinoma. Leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma (including cutaneous T-cell lymphoma or peripheral T-cell lymphoma), Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma, histiocytic lymphoma, and Burkitt's lymphoma; hematopoietic tumors of myeloid lineage, e.g., acute and chronic myeloid leukemia, myelodysplastic syndrome, myeloid leukemia, and promyelocytic leukemia; tumors of the central and peripheral nervous system, e.g., astrocytoma, neuroblastoma, glioma, and and Schwannoma; tumors of mesenchymal origin, e.g., fibrosarcoma, rhabdomyosarcoma, and osteosarcoma; other tumors, e.g., melanoma, xeroderma pigmentosum, keratoacanthoma, seminoma, follicular thyroid carcinoma, and teratocarcinoma; melanoma, unresectable stage III or IV malignant melanoma, squamous cell carcinoma, small cell lung cancer, non-small cell lung cancer, glioma, gastrointestinal cancer, renal cancer, ovarian cancer, liver cancer, colorectal cancer, endometrial cancer, kidney cancer, prostate cancer, thyroid cancer, neuroblastoma, pancreatic cancer, These include various cancers including, but not limited to, glioblastoma multiforme, cervical cancer, stomach cancer, bladder cancer, hepatocellular carcinoma, breast cancer, colon cancer, and head and neck cancer, retinoblastoma, gastric cancer, germ cell tumors, bone cancer, bone tumors, adult malignant fibrous histiocytoma of bone; pediatric malignant fibrous histiocytoma of bone, sarcoma, childhood sarcoma; myelodysplastic syndromes; neuroblastoma; testicular germ cell tumors, intraocular melanoma, myelodysplastic syndromes; myelodysplastic / myeloproliferative disorders, synovial sarcoma.

[0024] In some embodiments, cancer is solid tumor.For example, cancer can be sarcoma and osteosarcoma, such as Kaposi's sarcoma, AIDS-related Kaposi's sarcoma, melanoma, particularly uveal melanoma, as well as cancer of head and neck, kidney, ovary, pancreas, prostate, thyroid, lung, esophagus, breast, particularly triple-negative breast cancer (TNBC), bladder, colorectal, liver and bile duct, uterus, appendix, and cervix, testicular cancer, digestive cancer, and endometrial and peritoneal cancer.Preferably, cancer can be sarcoma, melanoma, particularly uveal melanoma, as well as cancer of head and neck, kidney, ovary, pancreas, prostate, thyroid, lung, esophagus, breast, particularly (TNBC), bladder, colorectal, liver, cervix, and endometrial and peritoneal cancer.

[0025] In some embodiments, the cancer may be selected from the group consisting of leukemia, lymphoma, sarcoma, melanoma, and cancer of the head and neck, kidney, ovary, pancreas, prostate, thyroid, lung, esophagus, breast, bladder, brain, colorectal, liver, and cervix.

[0026] In some embodiments, the cancer is lung cancer, particularly non-small cell lung cancer, leukemia, particularly acute myeloid leukemia, chronic lymphocytic leukemia, lymphoma, particularly peripheral T-cell lymphoma, chronic myeloid leukemia, squamous cell carcinoma of the head and neck, advanced melanoma with BRAF mutations, colorectal cancer, gastrointestinal stromal tumors, breast cancer, particularly HER2 + The cancer may be selected from the group consisting of breast cancer, thyroid cancer, particularly advanced medullary thyroid cancer, kidney cancer, particularly renal cell carcinoma, prostate cancer, glioma, pancreatic cancer, particularly pancreatic neuroendocrine carcinoma, multiple myeloma, and liver cancer, particularly hepatocellular carcinoma.

[0027] In particular, the subject suffers from lung cancer. As used herein, the term "lung cancer" has its general meaning in the art and refers to a disease in lung tissue that involves uncontrolled cell proliferation that in some cases leads to metastasis. The majority of primary lung cancers are carcinomas of the lung that originate from epithelial cells. The main types of lung cancer are small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC). In certain embodiments, the subject suffers from non-small cell lung cancer. As used herein, the term "non-small cell lung cancer", also known as non-small cell lung cancer (NSCLC), refers to epithelial lung cancer other than small cell lung cancer (SCLC). There are three main subtypes: adenocarcinoma, squamous cell lung carcinoma, and large cell lung cancer. Other less common types of non-small cell lung cancer include pleomorphic, carcinoid tumor, salivary gland carcinoma, and unclassified carcinoma. Adenocarcinoma accounts for about 40% of lung cancers and is the most common type of lung cancer in people who have never smoked. Squamous cell carcinoma accounts for approximately 25% of lung cancers. It is more common in men than women and is more strongly correlated with tobacco smoking history than other types of lung cancer. There are at least four variants of squamous cell carcinoma of the lung: papillary, small cell, clear cell, and basaloid. Large cell lung carcinoma is a heterogeneous group of malignant neoplasms that originate from cancerous epithelial cells in the lung. Large cell lung carcinoma is a carcinoma that does not have the light microscopic features of small cell carcinoma, squamous cell carcinoma, or adenocarcinoma. NSCLC can be classified using the tumor-node-metastasis (TNM) staging system.See Spira J & Ettinger, DS Multidisciplinary management of lung cancer, N Engl J Med, 350:382-(2004) (hereinafter Spira); Greene FL, Page DL, Fleming ID, Fritz AG, Balch CM, Haller DG et al. (eds). AJCC Cancer Staging Manual. 6th edition. New York: Springer-Verlag, 2002:167-77 (hereinafter Greene); Sobin LH, Wittekind CH (eds). International Union Against Cancer. TNM classification of malignant tumours. 6th edition. New York: Wiley-Liss (2002) (hereinafter Sobin). Thus, in some embodiments, lung cancer can be stratified into any of the aforementioned stages (e.g., occult, stage 0, stage IA, stage IB, stage IIA, stage IIB, stage IIIA, stage IIIB, or stage IV). More specifically, the subject is affected with EGFR mutated NSCLC or ALK mutated NSLC as described above.

[0028] Non-limiting exemplary Ras inhibitors include, but are not limited to, DCAI disclosed by Maurer (Maurer et al., 2012), Kobe0065 and Kobe2602 disclosed by Shima (Shima et al., 2013), HBS3 (Patgiri et al., 2011), AIK-4 (Allinky), adagrasib, ARS-3248, AZD4785, and sotorasib. Preferably, the Ras inhibitor is sotorasib, also known as AMG-510, which is an acrylamide-derived KR1 inhibitor developed by Amgen and has the IUPAC name 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]-4-[(2S)-2-methyl-4-(prop-2-enoyl)piperazin-1-yl]-1H,2H-pyrido[2,3-d]pyrimidin-2-one.

[0029] In some embodiments, the farnesyltransferase inhibitor can be an antimetabolite, such as an analog of farnesol, farnesyl phosphate, farnesyl diphosphate, or substrate peptide.The farnesyltransferase inhibitor can also be a molecule with a different structure that can bind to the binding pocket of peptide substrate or farnesyl diphosphate.Alternatively, the farnesyltransferase inhibitor can be an allosteric inhibitor.

[0030] In some embodiments, the farnesyltransferase inhibitor can have any molecular structure. For example, it can be a peptidic substance, a peptidomimetic, or a non-peptidic small molecule substance. A peptidic substance is composed mostly of peptides. However, peptides can be conjugated to other molecular structures, such as biologically compatible organic polymers (e.g., polyethylene glycol (PEG), polyethyleneimine (PEI), hydroxypropylmethacrylamide (HPMA)), lipids, alkyl moieties, or other polypeptides. A peptidomimetic is a substance whose molecular structure mimics a peptide. A peptidomimetic can, for example, contain or be made from beta amino acids (monoamino acids), gamma amino acids (γ amino acids), or D-amino acids. A peptidomimetic can also be conjugated to other molecular structures, such as biologically compatible organic polymers. A peptidomimetic can also be a retro-inverse peptide. A small molecule substance is a molecule with a molecular weight of less than 1500 Da, preferably less than 1000 Da, and even more preferably less than 500 Da. Small molecule agents can also be conjugated to other molecular structures, such as biologically compatible organic polymers.

[0031] In some embodiments, the farnesyltransferase inhibitor is selected from the group consisting of R11577 (Zarnestra, tipifarnib), SCH66336 (lonafarnib), FTI-277, GGTI-298, BMS-214664, L-778, and L-123.

[0032] In some embodiments, the farnesyltransferase inhibitor of the present invention is tipifarnib.As used herein, the term "tipifarnib" refers to the FTase inhibitor (R)-6-[amino(4-chlorophenyl)(l-methyl-1H-imidazol-5-yl)methyl]-4-(3-chlorophenyl)-l-methyl-2(lH)-quinolinone (also identified as Rl15777), also known as Zarnestra (J&JPRD) and having the structure shown below. [ka]

[0033] Typically, the agent of the present invention is administered to the subject in the form of a pharmaceutical composition containing a pharmaceutically acceptable carrier.The pharmaceutically acceptable carrier that can be used in this composition includes, but is not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphoric acid, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of vegetable saturated fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and wool fat.When used in administering to a subject, the composition can be formulated for administration to a subject.The composition of the present invention can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, bucally, vaginally, or via an indwelling reservoir. As used herein, it includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. The sterile injectable form of the composition of the present invention can be an aqueous or oily suspension. These suspensions can be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be used are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fixed oils are conventionally used as a solvent or suspending medium. For this purpose, any bland fixed oil can be used, including synthetic mono- or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharma- ceutical-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions.These oil solutions or suspensions may also contain long-chain alcohol diluents or dispersants, such as carboxymethylcellulose, or similar dispersants commonly used in the formulation of pharma- ceutically acceptable dosage forms, including emulsions and suspensions. Other commonly used surfactants, such as Tween, Span, and other emulsifiers or bioavailability enhancers commonly used in the manufacture of pharma-ceutically acceptable solid, liquid, or other dosage forms, may also be used for formulation purposes. The compositions of the present invention may be orally administered in any orally acceptable dosage form, including, but not limited to, capsules, tablets, aqueous suspensions, or solutions. In the case of tablets for oral use, commonly used carriers include lactose and cornstarch. Lubricants, such as magnesium stearate, are also typically added. For oral administration in capsule form, useful diluents include, for example, lactose. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifiers and suspending agents. If desired, certain sweeteners, flavors, or colorants may also be added. Alternatively, the compositions of the present invention can be administered in the form of suppositories for rectal administration. These can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature, and therefore dissolves in the rectum to release the drug. Such materials include cocoa butter, beeswax, and polyethylene glycol. The compositions of the present invention can also be administered topically, especially when the target of treatment includes areas or organs that are easily accessible by topical application, including diseases of the eye, skin, or lower intestinal tract. Suitable topical formulations are easily prepared for each of these areas or organs. For topical application, the compositions can be formulated into a suitable ointment containing the active ingredient suspended or dissolved in one or more carriers. Carriers for topical administration of the compounds of the present invention include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compounds, emulsifying wax, and water.Alternatively, the compositions can be formulated into a suitable lotion or cream containing the active ingredient suspended or dissolved in one or more pharma- ceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol, and water. Topical application for the lower intestinal tract can be with a rectal suppository (see above) or a suitable enema formulation. Patches can also be used. The compositions of the present invention can also be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well known in the art of pharmaceutical formulation and can be prepared as a solution in saline using benzyl alcohol or other suitable preservatives, absorption enhancers to enhance bioavailability, fluorocarbons, and / or other conventional solubilizing or dispersing agents. For example, the antibody present in the pharmaceutical composition of the present invention can be provided at a concentration of 10 mg / mL in either 100 mg (10 mL) or 500 mg (50 mL) single-use vials. The product is formulated for intravenous administration in 9.0 mg / mL sodium chloride, 7.35 mg / mL sodium citrate dihydrate, 0.7 mg / mL polysorbate 80, and sterile water for injection. The pH is adjusted to 6.5. An example of a suitable dosage range for the antibody in the pharmaceutical composition of the invention is about 1 mg / m. 2 ~500mg / m 2 However, it is understood that these plans are exemplary and that optimal plans and regimens can be adapted taking into account the affinity and tolerability of the particular antibody in the pharmaceutical composition, which needs to be determined in clinical trials. Pharmaceutical compositions of the present invention for injection (e.g., intramuscular, intravenous) can be prepared to contain sterile buffered water (e.g., 1 ml for intramuscular) and about 1 ng to about 100 mg, e.g., about 50 ng to about 30 mg, or more preferably about 5 mg to about 25 mg of the inhibitor of the present invention.

[0034] A further object of the present invention relates to a pharmaceutical composition or kit (kit of parts) comprising a farnesyltransferase inhibitor and a Ras inhibitor, especially for use in the treatment of cancer.

[0035] The present invention is further illustrated by the following figures and examples, which should not, however, be construed in any way as limiting the scope of the present invention. EXAMPLES

[0036] Example 1 method: H23 cells (KRas G12C) were seeded in 6-well plates and treated or not with sotorasib (1 μM), tipifarnib (1 μM), or their combination. Medium was changed twice a week, and cell culture density was monitored by an Incucyte® live cell analysis system.

[0037] result: Although sotorasib and tipifarnib did not show significant antitumor effects against H23 cells when used alone, the combination potently induced cell death, suggesting synergy between these agents (Figures 1A-B).

[0038] Example 2 Methods: H23 and Calu-1 KRAS(G12C) mutant non-small cell lung cancer (NSCLC) cell lines were transduced with the FUCCI (Fluorescent Ubiquitination-Based Cell Cycle Indicator) system and response / relapse to sotorasib (1 μM) or sotorasib (1 μM) + tipifarnib (1 μM) was monitored for 50 days by Incucyte® measuring total cell numbers or cell cycle kinetics.

[0039] Results: After an initial response to 1 μM sotorasib, H23 and Calu-1 cells gave rise to resistant proliferative clones (RPCs) that coincided with a gradual increase in cell cycle kinetics (Figures 2A, 2B, 2C, and 2D). Addition of 1 μM tipifarnib abrogated the development of RPCs by strongly affecting cell cycle kinetics, leading to cell death (Figures 2A, 2B, 2C, and 2D).

[0040] References Throughout this application, various references describe the state of the art to which this invention pertains. The disclosures of these references are incorporated herein by reference into this disclosure.

Claims

1. A method for treating cancer in a subject requiring treatment, comprising administering to the subject a therapeutically effective combination comprising a Ras inhibitor and a farnesyltransferase inhibitor.

2. A method for delaying and / or suppressing the development of cancer resistant to a Ras inhibitor in a subject, comprising administering a therapeutically effective amount of a Ras inhibitor to the subject in combination with a farnesyltransferase inhibitor.

3. A method for treating cancer resistant to Ras inhibitors in a subject requiring treatment, comprising administering a therapeutically effective amount of farnesyltransferase inhibitor to the subject.

4. A method for suppressing resistance to a Ras inhibitor administered to a subject suffering from cancer, comprising administering a therapeutically effective amount of a farnesyltransferase inhibitor to the subject.

5. The use of farnesyltransferase inhibitors to inhibit or suppress the development of cancer recurrence and / or acquired resistance to treatment with Ras inhibitors by inhibiting or suppressing the proliferation of persistent cancer cells.

6. The method according to any one of claims 1 to 5, wherein the patient has mutated Ras cancer.

7. The method according to claim 6, wherein the patient has a G12C KRAS mutation.

8. The method according to any one of claims 1 to 5, wherein the Ras inhibitor is sotrasib.

9. The method according to any one of claims 1 to 5, wherein the cancer is selected from the group consisting of leukemia, lymphoma, sarcoma, melanoma, and cancers of the head and neck, kidney, ovary, pancreas, prostate, thyroid, lung, esophagus, breast, bladder, brain, colorectal, liver, and cervix.

10. The method according to claim 9, wherein the subject is suffering from non-small cell lung cancer.

11. The method according to any one of claims 1 to 5, wherein the farnesyltransferase inhibitor is tipifarnib.

12. A pharmaceutical composition or kit (parts kit) containing a farnesyltransferase inhibitor and a Ras inhibitor, particularly for use in the treatment of cancer.

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