Drug for lung cancer that has acquired resistance to type I anticancer drugs

Abstract

The type 1 MET tyrosine kinase inhibitors capmatinib and tepotinib are effective against lung cancer having an MET exon 14 skipping mutation and have been approved as first-line drugs in Japan and the US, but it is expected that resistance to both of these drugs will be acquired. However, there were no prospects for drugs that could be used next.　Lung cancer therapeutics having as an active ingredient any compound of formula (1) and formula (2) can overcome resistance to capmatinib and tepotinib and manifest an effect as an inhibitor against lung cancer having an MET exon 14 skipping mutation in which the amino acids 1228 and 1230 of c-MET are mutated to an amino acid other than aspartic acid and tyrosine, respectively.

Description

[Technical field]

[0001] The present invention relates to a therapeutic agent for treating lung cancer in which a resistance secondary mutation has occurred against a type I MET tyrosine kinase inhibitor, by overcoming the resistance secondary mutation. More specifically, the present invention relates to a therapeutic agent for treating lung cancer in which a type I MET tyrosine kinase inhibitor has been administered to a lung cancer in which MET exon 14 skip has occurred, and a mutation has occurred in either aspartic acid (D) at position 1228 or tyrosine (Y) at position 1230 of c-Met, resulting in resistance to the type I MET tyrosine kinase inhibitor. [Background technology]

[0002] MET is a receptor for hepatocyte growth factor (HGF) with tyrosine kinase activity, and its genetic abnormalities have been reported in various types of cancer. MET exon 14 skipping mutation (METex14) is one of the MET genetic abnormalities and is found in approximately 3% of non-small cell lung cancer (NSCLC).

[0003] In 2020, two MET tyrosine kinase inhibitors (hereinafter referred to as "MET-TKIs"), capmatinib (US and Japan) and tepotinib (Japan), were approved by their respective health authorities as first-line drugs for patients with METex14-positive lung cancer due to their high efficacy.

[0004] Considering that the number of lung cancer deaths in Japan and the United States is approximately 220,000 per year, approximately 3% of these deaths, or approximately 5,000 people per year, could be candidates for MET-TKI treatment. These drugs are expected to become available in other countries, including the EU, in the future, and it is expected that more patients will benefit from MET-TKI.

[0005] However, it is known that although these MET-TKI treatments are effective for a while after the start of administration, resistance to the drugs inevitably develops within about a year. MET-TKI resistance mechanisms are broadly divided into two types: (1) resistance caused by the occurrence of a mutation (secondary mutation) in the binding site of MET-TKI, and (2) resistance caused by the activation of a bypass pathway other than MET. Of these, it has been reported that type (1) accounts for approximately 35% of MET-TKI resistance mechanisms. (Non-Patent Document 1)

[0006] Approved drugs such as capmatinib and tepotinib are classified as type I inhibitors that bind to the activated state of MET. Experiments using artificial tumor cell models with METex14 have reported that secondary mutations at Asp1228 (D1228: aspartic acid at position 1228) and Tyr1230 (Y1230: tyrosine at position 1230) frequently occur as a resistance mechanism to type I MET tyrosine kinase inhibitors (Non-Patent Document 2).

[0007] In fact, it has already been reported that when crizotinib, a type I tyrosine kinase inhibitor, is administered to lung cancer patients with METex14, resistance mutations occur at a high rate at D1228 and Y1230 (Non-Patent Document 3). In other words, mutations at D1228 and Y1230 can be used as an indicator that METex14-positive lung cancer patients have resistance to type I MET tyrosine kinase inhibitors. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] International Publication No. 2012 / 071321 [Patent Document 2] International Publication No. 2015 / 017607 [Non-patent literature]

[0009] [Non-Patent Document 1] Recondo, G. et al. 2019. JThorac Oncol [Non-Patent Document 2] Fujino,et al. 2019. JThorac Oncol [Non-Patent Document 3] Fujino,et al. 2019. Expert Opinion on Emerging Drugs Summary of the Invention [Problem to be solved by the invention]

[0010] These results indicate that even if METex14-positive cancer cells are initially treated with capmatinib or tepotinib, some cancer cells acquire secondary mutations at D1228 and Y1230 after a certain period of time, and that cancer cells that have acquired these secondary mutations still depend on MET signaling for survival and proliferation.

[0011] Therefore, it is important to use MET-TKIs that have inhibitory activity against secondary resistance mutations to control these tumors. However, drugs that can overcome these resistance mutations are not yet clear, even at the in vitro level. [Means for solving the problem]

[0012] The present invention has been conceived in view of the above problems, and provides a new class of MET-TKI for lung cancer that can overcome acquired resistance to capmatinib or tepotinib.

[0013] More specifically, the lung cancer therapeutic agent according to the present invention is MET_exon14_Skipping mutation Yes And c-Met 1228 Eye amino acid GaA Spartic Acid From (D), at least alanine (A) or tyrosine (Y) Mutated to In addition, the 1230th amino acid is mutated from tyrosine (Y) to any of the following amino acids: cysteine ​​(C), aspartic acid (D), histidine (H), asparagine (N), or serine (S).The present invention relates to a lung cancer treatment drug for lung cancer (lung cancer that has acquired resistance to type I MET-TKI), the active ingredient of which is a compound represented by formula (1) or (2).

[0014] [ka]

[0015] [ka]

[0016] The present invention may also be said to be a treatment for lung cancer having Met_exon14_skipping, in which the 1228th and 1230th amino acids of c-Met are mutated to amino acids other than aspartic acid and tyrosine, respectively, using a drug having as an active ingredient either a compound of formula (1) or (2). Effect of the Invention

[0017] The lung cancer therapeutic agent containing the compound of formula (1) or (2) according to the present invention as an active ingredient is effective in inhibiting cell proliferation of lung cancer having a METex14 mutation in which the 1228th and 1230th amino acids of c-Met are mutated to amino acids other than aspartic acid and tyrosine, respectively, i.e., lung cancer that is METex14 positive and has acquired resistance to type I MET-TKI.

[0018] In addition, it is said that the incidence of lung cancer caused by METex14 is equivalent to that of lung cancer caused by ALK fusion gene mutations. Regarding ALK fusion gene mutations, five types of ALK tyrosine kinase inhibitors (ALK-TKIs) with different mechanisms have already been developed and marketed, and even if resistance to the first ALK-TKI is acquired, resistance can be overcome by switching to another ALK-TKI, and prognosis can be improved. Therefore, there is a high expectation that a therapeutic drug that overcomes resistance in lung cancer that is METex14 positive and has acquired resistance to type I MET-TKIs will be commercially successful. [Brief description of the drawings]

[0019] [Figure 1] FIG. 1 shows the relationship between drug concentration and cell viability when cabozantinib was administered to Ba / F3 cells in which aspartic acid (D) at position 1228 of c-Met was mutated to asparagine (N). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0020] The lung cancer therapeutic agent according to the present invention will be described below with reference to the drawings and examples. Note that the following description is merely an example of one embodiment of the present invention and an example, and the present invention is not limited to the following description. The following description can be modified within the scope of the present invention.

[0021] The lung cancer therapeutic agent according to the present invention contains compounds represented by formula (1) and formula (2) as active ingredients. These compounds were found by a method of finding drugs exhibiting inhibitory activity from a drug library with various mechanisms of action using a cell model having a secondary mutation predicted to appear as a resistance mutation to type I MET-TKI. Note that type I MET-TKI is a drug that exhibits inhibitory activity by binding to the DFG-in structure of tyrosine kinase.

[0022] The substance represented by formula (1) is foretinib (Patent Document 1: International Publication No. 2012 / 071321), and the substance represented by formula (2) is CFP-40783 (Patent Document 2: International Publication No. 2015 / 017607). These drugs are type II MET-TKIs. Type II MET-TKIs are drugs that exhibit inhibitory effects by binding to the DFG-out structure of tyrosine kinase. In other words, they have a different mechanism of action from type I MET-TKIs, and are highly likely to overcome resistance to type I MET-TKIs.

[0023] As shown in the Examples below, these substances showed inhibitory effects against resistant strains caused by capmatinib and tepotinib. Therefore, it is believed that these substances are effective in overcoming resistance and suppressing cancer cell proliferation in lung cancers that are METex14 mutation-positive and have acquired resistance to type I MET-TKI (MET tyrosine kinase inhibitor).

[0024] In other words, lung cancer that is METex14 mutation-positive and has acquired resistance to type I MET-TKI can be treated using a drug containing a compound of formula (1) or (2) as an active ingredient.

[0025] According to Patent Document 1, foretinib of formula (1) can be administered to humans as a pharma- ceutically acceptable salt. For example, it can be administered as a free base. Specifically, foretinib can be administered at a dose of, for example, about 7.5 mg, 15.0 mg, 30.0 mg, and / or 45.0 mg per day.

[0026] Foretinib may also be provided in the form of tablets. In some cases, the tablets may contain hypromellose, sodium lauryl sulfate, lactose monohydrate, microcrystalline cellulose, croscarmellose sodium, and magnesium stearate. Some tablets may also contain hypromellose, titanium dioxide, and polyethylene glycol. Additionally, the tablets may contain solysorbate 80 and yellow iron oxide.

[0027] Also, foretinib (also referred to in Patent Document 1 as N1-{3-fluoro-4-[(6-(methyloxy)-7-{[3-(4-morpholinyl)propyl]oxy}-4-quinolinyl)oxy]phenyl}-N1-(4-fluorophenyl)-1,1-cyclopropanedicarboxamide), together with pharma- ceutically acceptable salts and solvates thereof, have been disclosed and claimed in International Application No. PCT / US2004 / 031523, having an international filing date of September 24, 2004, as being useful as an inhibitor of cMET, particularly for the treatment of cancer, along with methods for its preparation.

[0028] Its international publication number is WO 2005 / 030140, and its international publication date is April 7, 2005. And, in Examples 25 (p. 193), 36 (pp. 202-203), 42 (p. 209), 43 (p. 209), and 44 (pp. 209-210), methods for preparing foretinib are described.

[0029] Foretinib can be prepared as described in International Application No. PCT / US2009 / 064341, which has an international filing date of November 13, 2008, and International Publication No. WO 2010 / 056960, which has an international publication date of May 20, 2010.

[0030] Foretinib can be prepared as described in International Application No. PCT / US2009 / 058276, which has an international filing date of September 25, 2009, and International Publication No. WO 2010 / 036831, which has an international publication date of April 1, 2010.

[0031] Also, according to Patent Document 2, CEP-40783 or its pharma- ceutical acceptable salt can be administered in a wide variety of oral and / or parenteral dosage forms. In one embodiment, the compound of the present invention is delivered orally. Parenteral administration can be by injection, that is, intravenous, intramuscular, intradermal, subcutaneous, intraduodenal, or intraperitoneal administration.

[0032] CEP-40783 or its pharmaceutically acceptable salt can also be administered intravenously or subcutaneously.CEP-40783 or its pharmaceutically acceptable salt can also be administered by inhalation, for example, intranasally, and can also be administered transdermally.CEP-40783 or its pharmaceutically acceptable salt can also be delivered rectally, by buccal tablet, or by insufflation.

[0033] It is within the skill of the art to determine the appropriate dosage of CEP-40783 or its pharmacologic acceptable salt for a particular situation.Generally, treatment is started with a smaller dosage that is less than the optimal dosage of the compound.The dosage is then increased by small increments until the optimal effect is reached under the circumstances.The total daily dosage can be divided as appropriate, and if desired, can be administered multiple times throughout the day.

[0034] For example, CEP-40783 or a salt thereof can be administered once to four times a day. A standard dose is about 1 mg to about 1,000 mg (e.g., about 5 mg to about 500 mg).

[0035] In some cases, the standard dose is about 1 mg to about 300 mg (e.g., about 5 mg to about 250 mg), and in yet other cases, the standard dose may be about 10 mg to 100 mg.

[0036] Also, CEP-40783 or a salt thereof is administered according to body weight. For example, CEP-40783 or a salt thereof may be administered in an amount of about 0.1 mg / kg to about 500 mg / kg (e.g., about 1 mg / kg to about 100 mg / kg, or about 5 mg / kg to about 75 mg / kg). Also, CEP-40783 or a salt thereof may be administered in an amount of about 10 mg / kg to about 55 mg / kg. CEP-40783 can be prepared using any number of different methods, for example, using the methods described in WO2013 / 0074633.

[0037] These disclosures regarding foretinib and CEP-40783 may be incorporated herein. EXAMPLES

[0038] A Ba / F3 cell model harboring secondary mutations (D1228A / E / G / N / Y / V and Y1230C / D / H / N / S) predicted to emerge as resistance mutations to type I MET-TKIs was generated using genetic engineering techniques.

[0039] <Establishment of Ba / F3 cells with MET exon14 skipping mutation> Ba / F3 cells are mouse-derived proB cells and usually require exogenous interleukin 3 (IL3) for their survival. However, when driver gene mutations are introduced exogenously using retroviruses or the like, it is known that they transform and show survival and proliferation depending only on the introduced driver gene mutations even without IL3.

[0040] First, a recombinant retrovirus with the MET exon14 skipping mutation was created and used to infect Ba / F3 cells.

[0041] IL3 was removed to establish Ba / F3 cells that show survival and proliferation depending only on the MET exon 14 skipping gene mutation. These are called METex14-Ba / F3 cells.

[0042] <Establishment of Ba / F3 cells resistant to capmatinib and tepotinib> Next, a resistant strain with resistance to type I MET-TKIs was created. The METex14-Ba / F3 cells established above were exposed to ENU (ethylnitrosourea) for 24 hours.

[0043] Capmatinib or tepotinib was administered respectively on a 96-well plate and cultured for about 3 weeks in their presence to create cell lines with mutations in the tyrosine kinase binding site of MET and thus resistant.

[0044] The kinase domain of the obtained resistant cell line was analyzed by Sanger sequencing to examine the type of secondary mutation, and cells with each mutation were established. As a result, METex14-Ba / F3 cells were obtained in which the 1228th aspartic acid (D) of c-Met in METex14-Ba / F3 cells was mutated to alanine (A), glutamic acid (E), glycine (G), histidine (H), asparagine (N), valine (V), or tyrosine (Y). The respective cell lines are simply called D1228A, D1228E, D1228G, D1228H, D1228N, D1228V, and D1228Y.

[0045] We also obtained METex14-Ba / F3 cells in which tyrosine (Y) at position 1230 of c-Met was mutated to cysteine ​​(C), aspartic acid (D), histidine (H), asparagine (N), or serine (S). The respective cell lines are simply called Y1230C, Y1230D, Y1230H, Y1230N, and Y1230S. These cells with mutations at positions D1228 and Y1230 are also called "sample cells." These may also be called "METex14-Ba / F3-resistant cells."

[0046] These cells are model cells resistant to type I MET-TKIs. They may also be said to be cells in which the 1228th and 1230th amino acids of c-Met have been mutated to amino acids other than aspartic acid (D) and tyrosine (Y). Furthermore, they are model cells that have METex14 mutations and have acquired resistance to capmatinib or tepotinib.

[0047] These model cells were administered 300 drugs with various mechanisms of action registered in the drug library (hereafter referred to as "sample drugs") to search for drugs that could overcome resistance and act as MET-TKIs. The specific procedure was as follows:

[0048] On day 1, four types of Ba / F3 cells, namely normal Ba / F3 cells, METex14-Ba / F3 cells (without secondary mutation), and two types of METex14-Ba / F3 resistant cells (with secondary mutation), D1228A and D1228Y, were seeded in a 96-well plate at 3,000 cells each. IL3 was also added to the normal Ba-F3 cells.

[0049] On the second day, all 300 drugs were administered at a concentration of 100 nM. In addition, DMSO was administered to one well per plate as a control. This well was called the "control well."

[0050] On the fifth day (72 hours after drug administration), 10 μl of Cell Counting Kit 8 was administered to each well, and 3 hours later, the absorbance at 450 nM was directly measured using a plate reader to determine the number of viable cells.

[0051] Candidate drugs were selected from those that reduced viability to 50% or less in three types of cells, METex14-Ba / F3 cells, D1228A, and D1228Y (both of which are METex14-Ba / F3-resistant cells), compared with control wells administered DMSO. Drugs that were also effective on normal Ba / F3 cells were excluded, as they were determined not to have a specific inhibitory effect on the MET signal. IC50 and viability were calculated as follows.

[0052] The survival rate for each sample drug was calculated as the ratio (percentage) of the number of live cells in each well to the number of live cells in the control well, and a curve of survival rate against the concentration of the sample drug was created. An example of the graph is shown in Figure 1.

[0053] Figure 1 shows a graph of drug concentration (nM) vs. cell viability (%) when cabozantinib was administered to sample cells in which the 1228th aspartic acid (D) of c-Met was mutated to asparagine (N). The graph is labeled "METex14+D1228N-Ba / F3 cells vs. cabozantinib." The horizontal axis is the sample drug concentration (nM) and the vertical axis is the viability (%). It can be seen that the sample drug concentration is approximately 37.04nM and the cell viability is 50%. Based on this graph, the drug concentration at which the survival rate is 50% (IC50) was calculated for each sample drug. Table 1 shows examples of the results for sample drugs that were able to overcome resistance to secondary mutations and those that were not.

[0054] [Table 1]

[0055] Referring to Table 1, the conclusion was that only two drugs, CEP-40783 and foretinib, were able to overcome resistance against all mutations predicted as secondary mutations.

[0056] Referring to Table 1, on the left side are the METex14 parent strain (METex14-Ba / F3 cells), METex14-Ba / F3 mutant cells in which aspartic acid (D) at position 1228 has been mutated to alanine (A), glutamic acid (E), glycine (G), histidine (H), asparagine (N), valine (V), or tyrosine (Y), and METex14-Ba / F3 mutant cells in which tyrosine (Y) at position 1230 has been mutated to cysteine ​​(C), aspartic acid (D), histidine (H), asparagine (N), or serine (S).

[0057] As mentioned above, each cell line is also called D1228A, D1228E, D1228G, D1228H, D1228N, D1228V, D1228Y, Y1230C, Y1230D, Y1230H, Y1230N, and Y1230S.

[0058] The first and second rows show the IC50 concentrations when capmatinib and tepotinib were administered as type I MET-TKIs. The third and fourth rows show the results when CEP-40783 and foretinib were administered as type II MET-TKIs. The fourth and fifth rows show the results when cabozantinib and merestinib were administered as type II MET-TKIs.

[0059] Type I MET-TKIs (capmatinib and tepotinib) effectively inhibited c-Met against the parent cell line of METex14, with IC50 values ​​of 0.6 nM and 3.0 nM.

[0060] However, for METex14-Ba / F3 mutant cells with secondary mutations, the IC50 was 100nM or higher for all secondary mutations. In particular, the IC50 of tepotinib was higher than 1000nM for all METex14-Ba / F3 mutant cells. In other words, these type I MET-TKIs did not have an inhibitory effect on secondary mutant cells. In other words, it can be said that the secondary mutant METex14-Ba / F3 mutant cells had acquired resistance to these type I sample drugs.

[0061] Cabozantinib and merestinib, which are type II MET-TKIs, had low IC50 values ​​of 30 nM or less when tyrosine (Y) at position 1230 was mutated. In other words, it can be said that they overcame resistance and exerted c-Met inhibitory effects. However, when aspartic acid (D) at position 1228 was mutated, IC50 values ​​were often greater than 30 nM.

[0062] On the other hand, type II MET-TKIs CEP-40783 and foretinib showed excellent results, with IC50 values ​​of 30 nM or less and in most cases 10 nM or less, for both 1228 and 1230 mutations. In other words, it was concluded that these two drugs overcome resistance and exert MET tyrosine kinase inhibitory effects in cells expressing METex14 and with mutations at amino acids 1228 and 1230.

[0063] Based on the above, it can be said that these two sample drugs can be used as drugs that can overcome resistance and continue to be used after type I MET-TKIs become ineffective due to resistance.

[0064] In other words, it is possible to use these drugs to treat lung cancer that expresses METex14 and has mutations at amino acids 1228 and 1230 (lung cancer that has acquired resistance to type I MET-TKIs). [Industrial Applicability]

[0065] The lung cancer therapeutic drug of the present invention is effective against lung cancer in which METex14 is expressed and which has mutations at amino acids 1228 and 1230 (lung cancer that has acquired resistance to type I MET-TKI), and is expected to have an effect of suppressing the proliferation of cancer cells in vivo.

Claims

[Claim 1] A therapeutic agent for lung cancer, comprising as an active ingredient a compound of formula (1) or (2), for use in lung cancer having a MET_exon14_Skipping mutation, in which the 1228th amino acid of c-Met is mutated from aspartic acid (D) to at least alanine (A) or tyrosine (Y), and the 1230th amino acid is mutated from tyrosine (Y) to any of cysteine ​​(C), aspartic acid (D), histidine (H), asparagine (N), and serine (S). 【Chemistry 100】 【Chemistry 101】

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