Pharmaceutical combination of EGFR kinase inhibitor and c-met kinase inhibitor for treating tumor diseases

Abstract

The present invention relates to a pharmaceutical combination of an EGFR kinase inhibitor and a c-Met kinase inhibitor, and a use thereof in the preparation of a drug for treating tumor diseases. Specifically, provided is a pharmaceutical combination of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a compound of formula (II) or a pharmaceutically acceptable salt thereof. The combination of the two compounds or pharmaceutically acceptable salts thereof shows stronger anti-tumor activity, has no obvious toxic side effects, and has wide prospects for application.

Description

Drug combination of EGFR kinase inhibitor and c-Met kinase inhibitor for treating tumor diseases Technical Field

[0001] The present invention belongs to the field of medicine, and specifically relates to a drug combination of an EGFR kinase inhibitor and a c-Met kinase inhibitor, and use of the drug combination in preparing a drug for treating tumor diseases. Background Art

[0002] Lung cancer is the most common malignant tumor in China and worldwide. The latest global cancer data for 2020 released by the World Health Organization's International Agency for Research on Cancer (IARC) shows that there were 2.2 million new cases of lung cancer worldwide, ranking second in incidence and first in mortality. In China, lung cancer has the highest incidence and mortality rates of all malignant tumors, and both rates increase with age. Based on histopathology, lung cancer can be divided into non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC). NSCLC accounts for approximately 85% of all lung cancers. Approximately 75% of NSCLC patients are diagnosed in the advanced stage, with a very low five-year survival rate and a poor prognosis. For unresectable advanced stage lung cancer, despite some progress in traditional radiotherapy and chemotherapy, the treatment efficacy remains low. Advanced or metastatic lung cancer remains a deadly disease with significant unmet medical needs.

[0003] The EGFR (Epidermal Growth Factor Receptor) gene is one of the most common driver genes in NSCLC, with approximately 50% of Chinese NSCLC patients harboring EGFR gene mutations (J Thorac Oncol, 2014, 9(9):e70). EGFR tyrosine kinase inhibitors (TKIs), the most commonly used targeted drugs for treatment, have made breakthrough progress and have become the standard first-line treatment for patients with EGFR mutation-positive advanced NSCLC.

[0004] Although EGFR kinase inhibitors have become the standard of care for first-line treatment of patients with EGFR-mutated NSCLC, resistance can develop after 9-14 months of treatment, driven by the emergence of new EGFR or other gene mutations or amplifications. Existing data suggest that MET (Mesenchymal Epithelial Transition factor) gene amplification may contribute to acquired resistance to EGFR kinase inhibitors. MET abnormalities are present in some EGFR mutation-positive NSCLC patients who develop acquired resistance to EGFR kinase inhibitors. Therefore, the combination of EGFR kinase inhibitors with c-Met kinase inhibitors has potential application in the treatment of NSCLC patients whose resistance is due to MET gene amplification or MET protein overexpression.

[0005] WO2016 / 082713A1 discloses a class of 2-aminopyrimidine compounds that can potently inhibit the kinase activity of EGFR T790M resistance mutations and EGFR sensitive mutations, and show significant in vitro and in vivo anti-tumor activity against tumors driven by EGFR mutations. Among them, the compound N-((5-((5-chloro-4-((naphthalen-2-yl)amino))pyrimidin-2-yl)amino)-2-((N-methyl-N-dimethylaminoethyl)amino)-4-methoxyphenyl)acrylamide has the structure shown in formula (I):

[0006] WO2020 / 156453A1 discloses a class of tricyclic compounds containing pyrimidine groups, which can efficiently and selectively inhibit the activity of c-Met kinase, can be used to treat diseases mediated by MET gene abnormalities, and specifically discloses the compound shown in formula (II).

[0007] Summary of the Invention

[0008] In a first aspect, the present invention provides a drug combination for treating tumor diseases, comprising (i) an EGFR kinase inhibitor; and (ii) a c-Met kinase inhibitor.

[0009] In some embodiments, the EGFR kinase inhibitor is selected from a compound of formula (I) or a pharmaceutically acceptable salt thereof, gefitinib, erlotinib, icotinib, lapatinib, afatinib, dacomitinib, neratinib, osimertinib, ametinib, nocetinib, vometinib, rociletinib, avitinib, befortinib, lazertinib, CK-101, ASP8273, nazartinib or a pharmaceutically acceptable salt thereof.

[0010] In some embodiments, the c-Met kinase inhibitor is selected from a compound of formula (II) or a pharmaceutically acceptable salt thereof, crizotinib, cabozantinib, capmatinib, tepotinib, savolitinib, gumetinib, sitravatinib, EMD-1214063, elzovantinib, bozitinib, XL-092, AL-2846, TAS-115, AMG-337, HS-10241, TQ-B3139, ABN-401 or a pharmaceutically acceptable salt thereof.

[0011] In an exemplary embodiment, the EGFR kinase inhibitor is selected from the compound of formula (I) or a pharmaceutically acceptable salt thereof, and the c-Met kinase inhibitor is selected from the compound of formula (II) or a pharmaceutically acceptable salt thereof.

[0012] The second aspect of the present invention provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in combination with a compound of formula (II) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating tumor diseases.

[0013] In some embodiments, the tumor disease is selected from breast cancer, ovarian cancer, prostate cancer, melanoma, brain tumor, esophageal cancer, gastric cancer, liver cancer, pancreatic cancer, colorectal cancer, lung cancer, kidney cancer, skin cancer, glioblastoma, neuroblastoma, sarcoma, liposarcoma, osteochondroma, osteoma, osteosarcoma, seminoma, testicular tumor, uterine cancer, head and neck tumor, multiple myeloma, malignant lymphoma, polycythemia vera, leukemia, thyroid tumor, ureteral tumor, bladder cancer, gallbladder cancer, bile duct cancer or choriocarcinoma, preferably lung cancer, more preferably non-small cell lung cancer.

[0014] In some preferred embodiments, the non-small cell lung cancer is selected from squamous cell carcinoma or non-squamous cell carcinoma, preferably non-squamous cell carcinoma, wherein non-squamous cell carcinoma can be adenocarcinoma, large cell carcinoma and other subtypes of cell carcinoma.

[0015] In some preferred embodiments, the tumor disease is a tumor disease for which the patient is resistant to EGFR kinase inhibitors.

[0016] In some preferred embodiments, the tumor disease is a tumor disease containing EGFR mutation and / or MET abnormality, preferably a tumor disease containing one or more gene mutations of EGFR 19del, MET abnormality, EGFR L858R, and EGFR T790M.

[0017] In some preferred embodiments, the tumor disease is lung cancer that is resistant to EGFR kinase inhibitors.

[0018] In some preferred embodiments, the tumor disease is lung cancer containing EGFR mutation and / or MET abnormality, preferably lung cancer containing one or more gene mutations of EGFR 19del, MET abnormality, EGFR L858R, and EGFR T790M.

[0019] In some preferred embodiments, the tumor disease is non-small cell lung cancer in which the patient is resistant to EGFR kinase inhibitors.

[0020] In some preferred embodiments, the tumor disease is non-small cell lung cancer containing EGFR mutation and / or MET abnormality, more preferably non-small cell lung cancer containing one or more gene mutations of EGFR 19del, MET abnormality, EGFR L858R, and EGFR T790M.

[0021] In an exemplary embodiment, the tumor disease is non-small cell lung cancer that has progressed after previous treatment with an EGFR kinase inhibitor and has MET abnormalities.

[0022] In some embodiments, the single dose of the compound of formula (I) or a pharmaceutically acceptable salt thereof is selected from 40 to 320 mg, preferably 60 to 240 mg, and more preferably 80 to 160 mg.

[0023] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered at a frequency selected from once a day, twice a day, three times a day, or once every two days.

[0024] In some embodiments, the single administration dose of the compound of formula (II) or a pharmaceutically acceptable salt thereof is selected from 40 to 640 mg, preferably 40 to 320 mg, and more preferably 60 to 320 mg.

[0025] In some embodiments, the compound of formula (II) or a pharmaceutically acceptable salt thereof is administered at a frequency selected from once a day, twice a day, three times a day, or once every two days.

[0026] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof and the compound of formula (II) or a pharmaceutically acceptable salt thereof can be administered simultaneously or at different times.

[0027] The third aspect of the present invention further provides a pharmaceutical preparation comprising the drug combination according to the first aspect of the present invention and one or more pharmaceutically acceptable carriers.

[0028] As used herein, the term "pharmaceutically acceptable salt" refers to a base addition salt obtained by contacting a compound with a sufficient amount of a base in a pure solution or a suitable inert solvent when the compound contains a relatively acidic functional group; or an acid addition salt obtained by contacting a compound with a sufficient amount of an acid in a pure solution or a suitable inert solvent when the compound contains a relatively basic functional group. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts, or similar salts. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, bisulfate, hydroiodic acid, phosphorous acid, and the like; and organic acid salts such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid, as well as salts of amino acids such as arginine, and salts of organic acids such as glucuronic acid.

[0029] As used herein, the term "drug combination" includes fixed-dose combinations (in a single dosage form), drug combination packages for combined administration, and combination therapies. In the case of drug combination packages or combination therapies, the drugs may be administered simultaneously or separately at intervals. As used herein, the term "fixed-dose combination" refers to a single dosage form formulated to deliver a fixed amount of two or more drugs to a patient; exemplary examples include combination preparations and compound preparations. As used herein, the terms "combination preparation" or "combination preparation" refer to a drug product composed of several drug components; a "combination preparation" refers to a preparation composed of several drugs of different classes; a "combination preparation" refers to a preparation composed of several drugs of the same class, although other classes of drugs are also permitted. As used herein, the term "drug combination package" refers to a package consisting of two or more drug preparations with independent indications, usage, and dosages. As used herein, the term "combination therapy" refers to the administration of two or more drugs with independent indications, usage, and dosages to a patient to treat related diseases.

[0030] In the present invention, the term "EGFR 19del" indicates a deletion of exon 19 of EGFR; the term "EGFR L858R" indicates a mutation of amino acid 858 of the EGFR protein from L to R; and the term "EGFR T790M" indicates a mutation of amino acid 790 of the EGFR protein from T to M. "EGFR 19del" and "EGFR L858R" are common mutations of EGFR and are sensitive to first- to third-generation EGFR kinase inhibitors.

[0031] In the present invention, the term "MET alteration" includes MET gene exon 14 skipping mutation (MET 14 skipping), MET gene amplification, MET gene point mutation (mainly kinase domain mutation), MET gene fusion and MET protein overexpression. The incidence of MET 14 skipping in NSCLC patients is 0.9% to 4.0%; the incidence of primary MET gene amplification in NSCLC patients is 1% to 5%, the incidence of secondary MET gene amplification in patients with EGFR kinase inhibitor resistance is 5% to 50%, and the incidence in patients with anaplastic lymphoma kinase inhibitor resistance is about 13%; the incidence of MET protein overexpression in NSCLC patients is 13.7% to 63.7%. See Chinese Journal of Pathology, 2022, 51(11): 1094-1103.

[0032] Beneficial effects of the invention:

[0033] The present invention provides a drug combination of an EGFR kinase inhibitor and a c-Met kinase inhibitor, and their use in the preparation of a medicament for treating tumor diseases. Specifically, a drug combination of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a compound of formula (II) or a pharmaceutically acceptable salt thereof is provided. The combination of the two exhibits enhanced anti-tumor activity without significant toxic side effects, and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] FIG1 is a curve showing changes in body weight of mice during drug administration in Example 1.

[0035] FIG2 is a curve showing the change of tumor volume in Example 1.

[0036] FIG3 is a curve showing the change of tumor volume in Example 2.

[0037] Figure 4 is a tumor image (day 21) after the test drug in Example 2 acts on the LD1-0025-200662 human lung cancer NU / NU mouse tumor model. CR indicates complete tumor regression (remission). G1-G7 corresponds to the grouping and dosing information in Table 9 in Example 2.

[0038] FIG5 is a curve showing the weight change of mice during the administration process in Example 2. DETAILED DESCRIPTION

[0039] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended only to illustrate the present invention and are not intended to limit the scope of the present invention. Equivalent substitutions or corresponding improvements made to the present invention are still within the scope of protection of the present invention. The experimental methods in the following examples where specific conditions are not specified are generally based on conventional conditions or the conditions recommended by the manufacturer.

[0040] Example 1 Pharmacodynamic Study of the Test Drug in the Immunodeficient Mouse Xenotransplantation Model of the Engineered Cell Line NCI-H1975-HGF

[0041] 1. Purpose of the experiment

[0042] Verify the efficacy of the test drug in the immunodeficient mouse xenotransplantation model of the engineered cell line NCI-H1975-HGF.

[0043] 2. Experimental Materials

[0044] 2.1 Cells

[0045] The cell source information is shown in Table 1.

[0046] Table 1 Cell information

[0047] 2.2 Experimental animals

[0048] B-NDG mice, female, 8 weeks old, weighing 18-22 g, were purchased from Biocytogen.

[0049] 2.3 Test drugs

[0050] Vehicle: 20% solution / 80% 100mM sodium acetate buffer pH 5.5;

[0051] ASK-MET: The hydrochloride salt of the compound represented by formula (II), prepared in 20% solutol / 80% 100 mM sodium acetate buffer, pH 5.5, at a concentration of 0.2 mg / mL;

[0052] ASK-EGFR: a compound represented by formula (I), prepared with 0.5% sodium carboxymethylcellulose solution (0.5% CMC-Na), concentration: 2 mg / mL.

[0053] Solutol: polyethylene glycol-15 hydroxystearate;

[0054] 100mM sodium acetate buffer pH 5.5: 100mM sodium acetate buffer pH 5.5;

[0055] 20% solutol / 80% 100mM sodium acetate buffer pH 5.5 means the volume ratio of solutol to 100mM sodium acetate buffer pH 5.5 in the drug solution is 20:80, the same below.

[0056] 3. Experimental Methods

[0057] 1) Resuscitate NCI-H1975-HGF, culture in vitro, and obtain 1.5*10 7 cell.

[0058] 2) Fifteen 8-10 week-old immunodeficient mice were fed and adapted for one week and weighed.

[0059] 3) See Table 2 for inoculation conditions.

[0060] Table 2 Vaccination information Note: sc: subcutaneous injection.

[0061] 4) After inoculation, tumor volume and body weight were measured twice a week. When the average tumor volume reached about 100 mm 3 The animals were randomly divided into four groups based on tumor volume and body weight, with three mice in each group. Dosing began immediately after grouping. The start date of dosing was considered day 0. Dosing and grouping information is shown in Table 3.

[0062] Table 3 Grouping and dosing information Note: po: oral administration; QD: once a day.

[0063] 5) Administer the drug on Day 0.

[0064] 6) After the drug administration is completed, the experiment is terminated.

[0065] 4. Observation and Data Collection

[0066] 4.1 Daily Observation

[0067] After inoculation, animals were observed for morbidity and mortality every working day. Daily observations included tumor growth and the effects of the drug on the experimental animals, such as changes in activity, food and water intake, weight loss, changes in hair and eye appearance, death, and other clinical symptoms.

[0068] 4.2 Weight Monitoring

[0069] The body weight of the experimental animals was measured once a week from inoculation to grouping, and twice a week after grouping.

[0070] 4.3 Tumor volume measurement

[0071] After inoculation and before grouping, when the tumor is visible, the tumor volume of the experimental animals is measured once a week. After inoculation and grouping, the tumor volume of the experimental animals is measured twice a week. Tumor volume measurement uses a two-way measurement method. First, use a vernier caliper to measure the long and short diameters of the tumor, and then use the formula TV = 1 / 2 × a × b 2 Calculate the tumor volume, where a is the long diameter of the tumor and b is the short diameter of the tumor.

[0072] 5. Experimental endpoint data analysis method

[0073] At the end of the experiment, the following analysis methods will be used for data analysis (Table 4).

[0074] Table 4 Data analysis methods

[0075] 6. Data Statistical Analysis

[0076] For experiments with two groups, T-test analysis was used. For comparisons of three or more groups, one-way ANOVA was used. For comparisons of potential synergistic effects, two-way ANOVA was used. A p-value of less than 0.05 was considered significant.

[0077] 7. Experimental Results

[0078] In this experiment, the mice showed no obvious symptoms and showed no significant changes in overall body weight. Among the drug-treated groups, only the combination-treated group experienced weight loss, but the individual weight loss did not exceed 20%. The experiment ended on Day 17, and data from Day 15 were used for analysis. Body weight data, tumor volume data, and tumor growth inhibition for each group are shown in Figures 1 and 2, Tables 5, 6, and 7.

[0079] Table 5 Body weight changes in each group

[0080] Table 6 Average tumor volume data of each group in the experiment

[0081] Table 7 Tumor inhibition effect of the tested drugs in each group Note: a Mean ± standard error; P value: drug-treated groups (G2-G4) vs. solvent control group (G1), statistically analyzed by One-Way ANOVA method.

[0082] 8. Experimental Summary and Discussion

[0083] In group G2, ASK-MET-2 mg / kg, poQD administration, on day 15, the average tumor volume was 940.03 ± 159.86 mm 3 , which was smaller than the average tumor volume of the control group (1275.47±152.44mm 3 ), TGI (%) of this group was 28.48%, compared with the control group, p>0.05.

[0084] In the G3 group, ASK-MET+ASK-EGFR-2+20 mg / kg, poQD administration on day 15, the average tumor volume was 192.22±45.80 mm3, which was smaller than the average tumor volume of the control group (1275.47±152.44 mm 3 ), TGI (%) of this group = 91.79%, compared with the control group p < 0.05.

[0085] In the G4 group, ASK-EGFR-20 mg / kg and poQD were administered for 15 days, with an average tumor volume of 589.86 ± 36.92 mm 3 , which was smaller than the average tumor volume of the control group (1275.47±152.44mm 3 ), TGI (%) of this group = 58.01%, compared with the control group p < 0.05.

[0086] In summary, the combined administration group ASK-MET+ASK-EGFR-2+20 mg / kg (QD) has a strong efficacy, and the combination of the two has a significant synergistic effect.

[0087] Example 2 Pharmacodynamic study of the test drug in a human lung cancer LD1-0025-200662 PDX tumor transplant model

[0088] 1. Purpose of the experiment

[0089] An in vivo xenograft tumor model was established by subcutaneously inoculating human lung cancer tumor LD1-0025-200662 in NU / NU mice, and the pharmacodynamics of the test drugs ASK-MET alone, ASK-EGFR alone, ASK-MET+ASK-EGFR combination, and RefT+ASK-EGFR combination therapy were evaluated in this model.

[0090] 2. Experimental Materials

[0091] 2.1 In vivo transplant tumor model

[0092] LD1-0025-200662 (see Table 8), human lung cancer tumor tissue, passaged to FP3+3, was used in this efficacy study;

[0093] Table 8 Transplanted tumor model

[0094] 2.2 Experimental animals

[0095] NU / NU mice, female, age: 42-62 days, weight 18-22 g, were purchased from Zhejiang Weitonglihua Laboratory Animal Technology Co., Ltd. (production license: SCXK (Zhejiang) 2019-0001).

[0096] 2.3 Test drugs

[0097] Vehicle: 20% solution / 80% 100mM sodium acetate buffer pH 5.5;

[0098] ASK-MET: hydrochloride of the compound represented by formula (II), prepared in 20% solutol / 80% 100 mM sodium acetate buffer, pH 5.5, concentrations: 0.1 mg / mL, 0.3 mg / mL;

[0099] ASK-EGFR: the compound represented by formula (I), prepared with 0.5% sodium carboxymethylcellulose solution (0.5% CMC-Na), concentration: 2 mg / mL;

[0100] RefT: The drug is prepared in 20% solutol / 80% 100 mM sodium acetate buffer, pH 5.5, at a concentration of 0.3 mg / mL. RefT is a small molecule MET kinase inhibitor approved for marketing by the USFDA.

[0101] 3. Experimental Methods

[0102] LD1-0025-200662 tumor tissue was inoculated subcutaneously into immunodeficient mice and observed until tumors formed and reached a volume of approximately 1000 mm. 3 The tumor tissue was peeled off and evenly cut into pieces of approximately 3 mm × 3 mm × 3 mm (approximately 50-90 mg) and inoculated subcutaneously into NU / NU mice. The mice were observed and the growth of the tumor was monitored. The average tumor volume of the tumor-bearing mice reached 193.40 mm in the group. 3 Random grouping and dosing were performed at the time of grouping. Tumor volume and body weight were measured for all tumor-bearing mice before grouping. Mice were randomly divided into groups based on tumor volume using a randomized block design. Mice within each block were then randomly assigned to treatment groups. Grouping was designated Day 0. Specific grouping and dosing information is shown in Table 9.

[0103] Table 9 Drug administration and treatment Note: po: oral administration; QD: once a day.

[0104] Dosage volume: Adjust the administration volume according to the weight of the tumor-bearing mice (0.1 mL / 10 g).

[0105] 4. Experimental Observation

[0106] Throughout the experimental process, the use and observation of experimental animals were conducted in accordance with AAALAC animal care guidelines. Following tumor tissue inoculation, the animals were observed daily, and any morbidity and mortality were recorded. Throughout the experimental process, all animals were monitored and recorded for behavior, food intake, water intake, weight changes, coat gloss, and other abnormalities.

[0107] 5. Evaluation Metrics

[0108] The main purpose is to detect the growth inhibitory effect or complete cure ability of the test drug on LD1-0025-200662 human lung cancer transplanted tumors in vivo.

[0109] 5.1 Tumor volume and tumor-bearing mouse weight measurement: Use a vernier caliper to measure twice a week. The tumor volume is calculated as V = 0.5 × a × b 2 , a, b represent the long diameter and wide diameter of the tumor, respectively.

[0110] 5.2 Tumor growth inhibition rate TGI (%) = [1-(T i -T0) / (V i -V0)]×100, where T i -T0>0; TGI(%)=[1-(T i -T0) / T0]×100, where T i -T0<0. T0 is the average tumor volume of the drug-treated group at the time of first administration, T i V0 is the average tumor volume of the vehicle control group (Vehicle) at the first dose, and V i It is the average tumor volume of the vehicle control group at a certain measurement after the start of drug administration.

[0111] 5.3 Relative tumor growth rate T / C (%): The calculation formula is as follows: T / C% = T RTV / C RTV ×100%(T RTV :RTV in treatment group; C RTV : negative control group RTV). The relative tumor volume (RTV) was calculated based on the tumor measurement results. The calculation formula is RTV = Vt / V0, where V0 is the average tumor volume measured at the time of group administration (i.e., d0), Vt is the average tumor volume at a certain measurement, and T RTV with C RTV Get data for the same day.

[0112] 5.4 Synergistic effect of combined drug administration: King's formula Q = E(a+b) / [Ea+Eb-Ea×Eb], Ea is the TGI of a single drug, and a Q value > 1 reflects that the combined use of drugs has a synergistic effect.

[0113] 5.5 After the experiment, the tumor was removed, weighed and photographed. weight % = T weight / V weight ×100%,T weight is the average tumor weight at the end of the compound group experiment, V weight The mean tumor weight of the vehicle control group at the end of the experiment.

[0114] 5.6 The body weight of tumor-bearing mice was measured twice a week, and the weight gain ratio of mice after drug administration was calculated: RCBW (%) = (BW i –BW0) / BW0×100, BW i BW0 is the average body weight at the time of first administration.

[0115] 6. Data Statistical Analysis

[0116] All data are expressed as Mean ± SEM, and one-way ANOVA LSD (L) test was used to compare the significant differences in tumor volume and weight among the groups. *P < 0.05 was considered to be a significant difference.

[0117] 7. Experimental Results

[0118] 7.1 Tumor volume and weight

[0119] The purpose of this study was to establish an in vivo LD1-0025-200662 PDX xenograft model by subcutaneously inoculating NU / NU mice with LD1-0025-200662 PDX tumors. The anti-tumor efficacy of the test drug in this model was then evaluated. Tumor volume changes, tumor images, and statistical analysis results for the LD1-0025-200662 PDX model in the treatment groups are shown in Figures 3 and 4, and Tables 10, 11, and 12.

[0120] The drug administration cycle of this efficacy experiment was 21 days, after which the entire efficacy test was terminated. On day 21, the average tumor volumes of tumor-bearing mice in the Vehicle group, ASK-EGFR_20mg / kg administration group, ASK-MET_1mg / kg administration group, ASK-MET_3mg / kg administration group, ASK-MET_1mg / kg+ASK-EGFR_20mg / kg administration group, ASK-MET_3mg / kg+ASK-EGFR_20mg / kg administration group, and RefT_3mg / kg+ASK-EGFR_20mg / kg administration group were 1261.93±184.86mm, respectively. 3 、1239.58±146.6mm 3 、116.01±35.11mm 3 、3.54±2.35mm 3 、32.67±11.56mm 3 、0.00±0.00mm 3 and 100.67±24.39mm 3 Compared with the vehicle group, the tumor growth inhibition rates (TGI) (%) of the ASK-EGFR_20mg / kg, ASK-MET_1mg / kg, ASK-MET_3mg / kg, ASK-MET_1mg / kg + ASK-EGFR_20mg / kg, ASK-MET_3mg / kg + ASK-EGFR_20mg / kg, and RefT_3mg / kg + ASK-EGFR_20mg / kg groups were 1.96%, 131.27%, 197.93%, 180.72%, 200.00%, and 140.79%, respectively. The above results indicate that, with the exception of the ASK-EGFR_20mg / kg group, all other treatment groups exhibited statistically significant tumor growth inhibition (P<0.001, and T / C%<40%). In addition, compared with the corresponding single-drug treatment groups, the ASK-MET_1mg / kg+ASK-EGFR_20mg / kg administration group, the ASK-MET_3mg / kg+ASK-EGFR_20mg / kg administration group and the RefT_3mg / kg+ASK-EGFR_20mg / kg administration group all showed more significant inhibitory effects on tumor growth (P<0.05), exhibiting a significant synergistic effect (Q>1).

[0121] On the 21st day after the experiment ended, visual observation showed that the rates of complete tumor regression in the vehicle group, ASK-EGFR_20mg / kg group, ASK-MET_1mg / kg group, ASK-MET_3mg / kg group, ASK-MET_1mg / kg + ASK-EGFR_20mg / kg group, ASK-MET_3mg / kg + ASK-EGFR_20mg / kg group, and RefT_3mg / kg + ASK-EGFR_20mg / kg group were 0.00%, 0.00%, 12.5%, 75%, 12.5%, 100%, and 0.00%, respectively. Subsequently, all tumor-bearing mice were euthanized, and the subcutaneous transplanted tumors were removed and weighed. The results of tumor weight and tumor volume were consistent.

[0122] Table 10 Pharmacodynamic study of test drugs in human lung cancer LD1-0025-200662 in vivo transplanted tumor model Note: N, number of animals in each treatment group. Compared with the vehicle group, significant differences were considered when *p<0.05, **p<0.01, ***p<0.001, and p<0.05 with a T / C% <40%. a: Percentage of mice in each treatment group with complete tumor resolution as determined by visual inspection on day 21 after treatment.

[0123] Table 11 Statistical analysis results of the combined drug group compared with the single drug group (Day 21) Note: N, number of animals in each treatment group; compared with ASK-EGFR, 20 mg / kg group, # p<0.05, ## p<0.01, ### p<0.001, p<0.05 were considered to be significantly different; compared with the ASK-MET, 1 mg / kg group, ▲ p<0.05, p<0.05 was considered to be significantly different; compared with the ASK-MET, 3 mg / kg group, ▼ p<0.05, p<0.05 is considered to be a significant difference.

[0124] Table 12 Tumor mass weight (g) (Mean ± SEM) Note: N, number of animals in each treatment group; compared with the Vehicle group, *p<0.05, **p<0.01, ***p<0.001, p<0.05 and T / C%<40% were considered to be significantly different.

[0125] 7.2 Changes in mouse body weight

[0126] The results of body weight changes and relative body weight changes in each treatment group are shown in Figure 5 and Table 13. During this experiment, no mice in any treatment group showed significant weight loss or other abnormal symptoms, indicating that the tumor-bearing mice had good tolerance to the test drug.

[0127] Table 13 Body weight changes of each treatment group and control group in the human lung cancer LD1-0025-200662 in vivo transplanted tumor model

[0128] 8. Experimental Summary and Discussion

[0129] According to the above experimental results: after treatment, the ASK-MET_1mg / kg dosing group, ASK-MET_3mg / kg dosing group, ASK-MET_1mg / kg+ASK-EGFR_20mg / kg dosing group, ASK-MET_3mg / kg+ASK-EGFR_20mg / kg dosing group and RefT_3mg / kg+ASK-EGFR_20mg / kg dosing group were all able to show statistically significant inhibition of the growth of subcutaneously transplanted tumors in mice with human lung cancer LD1-0025-200662. Furthermore, the ASK-MET 1mg / kg, ASK-MET 3mg / kg, ASK-MET 1mg / kg + ASK-EGFR 20mg / kg, and ASK-MET 3mg / kg + ASK-EGFR 20mg / kg treatment groups all induced complete tumor regression in the tumor-bearing mice to varying degrees. Furthermore, the anti-tumor efficacy of the ASK-MET 1mg / kg + ASK-EGFR 20mg / kg and ASK-MET 3mg / kg + ASK-EGFR 20mg / kg combination treatment groups was significantly superior to that of the corresponding monotherapy groups, demonstrating a significant synergistic effect. Furthermore, no significant abnormalities in the weight or behavior of the tumor-bearing mice were observed throughout the treatment process, indicating that the tumor-bearing mice tolerated the test drug well.

[0130] Example 3: An open, multicenter, phase I clinical trial evaluating the safety, tolerability, pharmacokinetic characteristics, and preliminary efficacy of ASKC202 tablets combined with ASK120067 tablets in patients with advanced solid tumors

[0131] Clinical trial registration number: CTR20220697; wherein ASKC202 is the ASK-MET in Examples 1 and 2, and ASK120067 is the ASK-EGFR in Examples 1 and 2.

[0132] Table 14 Experimental drugs and medication regimen

[0133] The AEs experienced by the four subjects enrolled in this study during the first treatment cycle were mainly laboratory abnormalities, and significant anti-tumor activity was initially observed in two subjects during the first tumor assessment. Therefore, the combination of ASKC202 tablets and ASK120067 tablets has broad prospects for the treatment of locally advanced or metastatic non-small cell lung cancer.

[0134] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above disclosure, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. A drug combination for treating tumor diseases, comprising (i) an EGFR kinase inhibitor; (ii) a c-Met kinase inhibitor, characterized in that: The EGFR kinase inhibitor is selected from the compound of formula (I) or a pharmaceutically acceptable salt thereof, and the c-Met kinase inhibitor is selected from the compound of formula (II) or a pharmaceutically acceptable salt thereof, 2. Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in combination with a compound of formula (II) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a tumor disease; 3. The pharmaceutical combination according to claim 1 or the use according to claim 2, characterized in that: The tumor disease is selected from breast cancer, ovarian cancer, prostate cancer, melanoma, brain tumor, esophageal cancer, gastric cancer, liver cancer, pancreatic cancer, colorectal cancer, lung cancer, kidney cancer, skin cancer, glioblastoma, neuroblastoma, sarcoma, liposarcoma, osteochondroma, osteoma, osteosarcoma, seminoma, testicular tumor, uterine cancer, head and neck tumor, multiple myeloma, malignant lymphoma, polycythemia vera, leukemia, thyroid tumor, ureteral tumor, bladder cancer, gallbladder cancer, bile duct cancer or choriocarcinoma, preferably lung cancer, more preferably non-small cell lung cancer.

4. The pharmaceutical combination according to claim 1 or the use according to claim 2, characterized in that: The tumor disease is a tumor disease in which the patient is resistant to EGFR kinase inhibitors; preferably, non-small cell lung cancer is resistant to EGFR kinase inhibitors.

5. The pharmaceutical combination according to claim 1 or the use according to claim 2, characterized in that: The tumor disease is a tumor disease containing EGFR mutation and / or MET abnormality; preferably non-small cell lung cancer containing EGFR mutation and / or MET abnormality; more preferably non-small cell lung cancer containing one or more of EGFR 19del, MET abnormality, EGFR L858R, and EGFR T790M.

6. The pharmaceutical combination according to claim 1 or the use according to claim 2, characterized in that: The tumor disease is non-small cell lung cancer that has progressed after previous treatment with EGFR kinase inhibitors and has MET abnormalities.

7. The pharmaceutical combination according to claim 1 or the use according to claim 2, characterized in that: The single administration dose of the compound of formula (I) or a pharmaceutically acceptable salt thereof is selected from 40 to 320 mg, preferably 60 to 240 mg, more preferably 80 to 160 mg; The frequency of administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof is selected from once a day, twice a day, three times a day or once every two days; The single administration dose of the compound of formula (II) or a pharmaceutically acceptable salt thereof is selected from 40 to 640 mg, preferably 40 to 320 mg, more preferably 60 to 320 mg; The administration frequency of the compound of formula (II) or a pharmaceutically acceptable salt thereof is selected from once a day, twice a day, three times a day or once every two days.

8. The pharmaceutical combination according to claim 1 or the use according to claim 2, characterized in that: The compound of formula (I) or a pharmaceutically acceptable salt thereof and the compound of formula (II) or a pharmaceutically acceptable salt thereof may be administered simultaneously or at different times.

9. A pharmaceutical preparation comprising the pharmaceutical combination of claim 1 and one or more pharmaceutically acceptable carriers.

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