Pharmaceutical combinations of EGFR kinase inhibitors and c-Met kinase inhibitors for treatment of neoplastic diseases

Abstract

The invention relates to a pharmaceutical composition of an EGFR (epidermal growth factor receptor) kinase inhibitor and a c-Met kinase inhibitor and application of the pharmaceutical composition in preparation of medicines for treating tumor diseases. Specifically, the invention provides a pharmaceutical composition of a compound shown as a formula (I) or a pharmaceutically acceptable salt thereof and a compound shown as a formula (II) or a pharmaceutically acceptable salt thereof, and the combination of the two compounds shows stronger anti-tumor activity, has no obvious toxic or side effect and has a wide application prospect.

Description

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 even in the world. The latest global cancer data for 2020 released by the World Health Organization's International Agency for Research on Cancer (IARC) shows that the number of new lung cancer cases in the world is 2.2 million, with the incidence ranking second and the mortality ranking first; in China, the incidence and mortality of lung cancer rank first among all malignant tumors, and the incidence and mortality increase with age. According to histopathology, lung cancer can be divided into non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC), of which NSCLC accounts for about 85% of all lung cancers. About 75% of NSCLC patients are in the middle and late stages at the time of diagnosis, with a very low 5-year survival rate and poor prognosis. For unresectable middle and late stage lung cancer, although traditional radiotherapy and chemotherapy have made some progress, the treatment efficiency is still low, and advanced or metastatic lung cancer is still a fatal disease with a large number of unmet medical needs.

[0003] EGFR (Epidermal Growth Factor Receptor) gene is one of the most common driver genes in NSCLC, and about 50% of Chinese NSCLC patients have EGFR gene mutations (J Thorac Oncol, 2014, 9(9): e70). As the most commonly used targeted drug for treatment, EGFR tyrosine kinase inhibitors (TKI) 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, patients develop resistance after 9-14 months of treatment, and the mechanisms include the emergence of new EGFR or other gene mutations or amplifications. Existing research data show that MET (Mesenchymal Epithelial Transition factor) gene amplification may be one of the causes of patients' acquired resistance to EGFR kinase inhibitors. MET abnormalities are present in some NSCLC patients who are EGFR mutation-positive and have acquired resistance to EGFR kinase inhibitors. Therefore, the combination of EGFR kinase inhibitors and c-Met kinase inhibitors has potential application value in the treatment of NSCLC patients with resistance caused by MET gene amplification or MET protein overexpression.

[0005] WO2016 / 082713A1 discloses a class of 2-aminopyrimidine compounds, which can effectively inhibit the kinase activity of EGFR T790M resistance mutation and EGFR sensitive mutation, and show significant in vitro and in vivo anti-tumor activity against tumors driven by EGFR mutations; wherein 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 a structure as shown in formula (I):

[0006]

[0007] WO2020 / 156453A1 discloses a class of tricyclic compounds containing pyrimidine groups, which can effectively and highly 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).

[0008] Summary of the invention

[0009] In a first aspect, the present invention provides a drug combination for treating tumor diseases, wherein the drug combination comprises (i) an EGFR kinase inhibitor; and (ii) a c-Met kinase inhibitor.

[0010] 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, nocitinib, vometinib, rociletinib, avitinib, befortinib, lazertinib, CK-101, ASP8273, nazartinib or a pharmaceutically acceptable salt thereof.

[0011] 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.

[0012] 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.

[0013]

[0014] 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.

[0015] 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.

[0016] 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 the non-squamous cell carcinoma may be adenocarcinoma, large cell carcinoma and other subtypes of cell carcinoma.

[0017] In some preferred embodiments, the tumor disease is a tumor disease for which the patient is resistant to taking EGFR kinase inhibitors.

[0018] 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.

[0019] In some preferred embodiments, the tumor disease is lung cancer that is resistant to EGFR kinase inhibitors.

[0020] 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.

[0021] In some preferred embodiments, the tumor disease is non-small cell lung cancer that is resistant to EGFR kinase inhibitors.

[0022] 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.

[0023] In an exemplary embodiment, the tumor disease is non-small cell lung cancer with disease progression after previous treatment with an EGFR kinase inhibitor and MET abnormality.

[0024] In some embodiments, the single dose of the compound of formula (I) or its pharmaceutically acceptable salt is selected from 40 to 320 mg, preferably 60 to 240 mg, and more preferably 80 to 160 mg.

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

[0026] 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.

[0027] In some embodiments, the compound of formula (II) or a pharmaceutically acceptable salt thereof is administered once a day, twice a day, three times a day, or once every two days.

[0028] 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.

[0029] The third aspect of the present invention further provides a pharmaceutical preparation, which comprises the drug combination described in the first aspect of the present invention and one or more pharmaceutically acceptable carriers.

[0030] In the present invention, the term "pharmaceutically acceptable salt" means that when the compound contains a relatively acidic functional group, a base addition salt can be obtained by contacting such compound with a sufficient amount of base in a pure solution or a suitable inert solvent; or when the compound contains a relatively basic functional group, an acid addition salt can be obtained by contacting such compound with a sufficient amount of acid in a pure solution or a suitable inert solvent. 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, hydrogen sulfate, 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, and the like; and also include salts of amino acids (such as arginine, etc.), and salts of organic acids such as glucuronic acid.

[0031] In the present invention, the term "combination" of drugs includes fixed-dose combinations of drugs (in a single dosage form), drug combination packages for combined administration, and also includes drug combination therapy; when it is a drug combination package or a drug combination therapy, each drug can be administered simultaneously or separately at a certain time interval. In the present invention, the term "fixed-dose combination" refers to a single dosage form prepared for delivering a certain amount of two or more drugs to a patient; exemplary ones include compound preparations and composite preparations. In the present invention, the term "compound preparation" or "compound preparation" both refer to drugs composed of several drug ingredients; wherein "compound preparation" refers to a preparation mixed with several different types of drugs; wherein "compound preparation" refers to a preparation composed of several drugs of the same category, and of course other types of drugs are also allowed. In the present invention, the term "drug combination package" refers to a package consisting of two or more drug preparations with independent indications and usage and dosage. In the present invention, the term "combination therapy" of drugs refers to the administration of two or more drugs with independent indications and usage and dosage to patients to treat related diseases.

[0032] In the present invention, the term "EGFR 19del" means that the 19th exon of EGFR is deleted; the term "EGFR L858R" means that the 858th amino acid of EGFR protein mutates from L to R; the term "EGFR T790M" means that the 790th amino acid of EGFR protein mutates from T to M. Among them, "EGFR 19del" and "EGFR L858R" are common mutations of EGFR, which are more sensitive to the 1st to 3rd generation EGFR kinase inhibitors.

[0033] 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 region 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 EGFR kinase inhibitor-resistant patients is 5% to 50%, and the incidence in anaplastic lymphoma kinase inhibitor-resistant patients 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.

[0034] Beneficial effects of the invention:

[0035] The present invention provides a drug combination of an EGFR kinase inhibitor and a c-Met kinase inhibitor, and use thereof in preparing a drug 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, and the combination of the two shows stronger anti-tumor activity and has no obvious toxic side effects, and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 The figure is the weight change curve of mice during the administration process in Example 1.

[0037] Figure 2 This is the tumor volume change curve in Example 1.

[0038] Figure 3 This is the tumor volume change curve in Example 2.

[0039] Figure 4 This is a tumor picture (day 21) after the test drug in Example 2 acted on the LD1-0025-200662 human lung cancer NU / NU mouse tumor model. CR means complete tumor regression (remission). G1-G7 corresponds to the grouping and dosing information in Table 9 in Example 2.

[0040] Figure 5 The figure is the weight change curve of mice during the administration process in Example 2. DETAILED DESCRIPTION

[0041] The present invention is further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. Equivalent replacements or corresponding improvements made to the present invention still fall within the scope of protection of the present invention. The experimental methods in the following examples where specific conditions are not specified are usually carried out under conventional conditions or under conditions recommended by the manufacturer.

[0042] Example 1 Pharmacodynamic study of the test drug in the immunodeficient mouse xenotransplantation model of the engineered cell line NCI-H1975-HGF

[0043] 1. Purpose of the experiment

[0044] Verify the efficacy of the test drug in the immunodeficient mouse xenograft model of the engineered cell line NCI-H1975-HGF.

[0045] 2. Experimental Materials

[0046] 2.1 Cells

[0047] The cell source information is shown in Table 1.

[0048] Table 1 Cell information

[0049] Cell Name source Cancer Type generation Required cell amount NCI-H1975-HGF KYBC Engineered cell lines 15-25 generations <![CDATA[1.5*10 7 ]]>

[0050] 2.2 Experimental animals

[0051] B-NDG mice, female, age: 8 weeks, weight 18-22 g, purchased from Biocytogen.

[0052] 2.3 Test drugs

[0053] Vehicle: 20% solutol / 80% 100mM sodium acetate buffer pH 5.5;

[0054] ASK-MET: hydrochloride of the compound represented by formula (II), prepared with 20% solutol / 80% 100 mM sodium acetate buffer pH 5.5, concentration: 0.2 mg / mL;

[0055] ASK-EGFR: a compound represented by formula (I), the drug was prepared using 0.5% sodium carboxymethylcellulose solution (0.5% CMC-Na), concentration: 2 mg / mL.

[0056] Solutol: Macrogol-15 Hydroxystearate;

[0057] 100mM sodium acetate buffer pH 5.5: 100mM sodium acetate buffer pH 5.5;

[0058] 20% solutol / 80% 100mM sodium acetate buffer pH 5.5 means that the volume ratio of solutol to 100mM sodium acetate buffer pH 5.5 in the drug solution is 20:80, the same below.

[0059] 3. Experimental Methods

[0060] 1) Resuscitation of NCI-H1975-HGF, in vitro culture, and obtaining 1.5*10 7 cell.

[0061] 2) Fifteen 8-10 week-old immunodeficient mice were fed adaptively for one week and weighed.

[0062] 3) See Table 2 for inoculation conditions.

[0063] Table 2 Vaccination information

[0064]

[0065] Note: sc: subcutaneous injection.

[0066] 4) After inoculation, tumor volume and body weight were measured twice a week. When the average tumor volume reached about 100 mm 3 The mice were randomly divided into 4 groups according to tumor volume and body weight, with 3 mice in each group. Drug administration began immediately after grouping. The start date of drug administration was regarded as day 0. The drug administration and grouping information are shown in Table 3.

[0067] Table 3 Grouping and dosing information

[0068]

[0069] Note: po: intragastric administration; QD: once a day.

[0070] 5) Administration was performed on Day 0.

[0071] 6) After the drug administration is completed, the experiment is terminated.

[0072] 4. Observation and Data Collection

[0073] 4.1 Daily Observation

[0074] After inoculation, the animals were observed for morbidity and mortality every working day. Daily observations included tumor growth and the effects of drugs on experimental animals, such as changes in activity, changes in food and water intake, weight loss, changes in hair and eye appearance, death, and other clinical symptoms.

[0075] 4.2 Weight Monitoring

[0076] The body weight of experimental animals was measured once a week from inoculation to grouping, and twice a week after grouping.

[0077] 4.3 Tumor volume measurement

[0078] 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. The tumor volume is measured using a two-way measurement method. First, the long and short diameters of the tumor are measured with a vernier caliper, and then the formula TV = 1 / 2 × a × b is used. 2 Calculate the tumor volume, where a is the long diameter of the tumor and b is the short diameter of the tumor.

[0079] 5. Experimental endpoint data analysis method

[0080] At the end of the experiment, the following analysis methods will be used for data analysis (Table 4).

[0081] Table 4 Data analysis methods

[0082]

[0083] 6. Statistical analysis

[0084] For experiments with two groups, T-Test analysis was used, and for comparisons of more than three groups, one-way ANOVA was used. For comparisons of potential synergistic effects, two-way ANOVA was used. A p value less than 0.05 was considered to be significantly different.

[0085] 7. Experimental Results

[0086] In this experiment, the experimental mice did not show obvious symptoms, and the overall body weight did not change significantly. Among the drug-treated groups, only the combined drug-treated group showed a decrease in weight, but the decrease in a single mouse did not exceed 20%. The experiment ended on Day 17, and the data from Day 15 were used for analysis. The weight data, tumor volume data, and tumor growth inhibition of each group are shown in Figure 1 , Figure 2 , Table 5, Table 6 and Table 7.

[0087] Table 5 Body weight changes in each group

[0088]

[0089] Table 6 Average tumor volume data of each group in the experiment

[0090]

[0091] Table 7 Tumor inhibition effect of each group of tested drugs

[0092]

[0093] Note: a Mean ± standard error; P value: drug-treated groups (G2-G4) vs solvent control group (G1), statistically analyzed by One-Way ANOVA method.

[0094] 8. Experimental Summary and Discussion

[0095] Group G2, ASK-MET-2 mg / kg, poQD administration, day 15, 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 = 28.48%, compared with the control group, p>0.05.

[0096] In the G3 group, ASK-MET+ASK-EGFR-2+20 mg / kg, the average tumor volume on the 15th day of poQD administration 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.

[0097] Group G4, ASK-EGFR-20 mg / kg, poQD administration, day 15, average tumor volume 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.

[0098] In summary, the combined administration group ASK-MET+ASK-EGFR-2+20mg / kg (QD) has a strong efficacy, and the combination of the two has a significant synergistic effect.

[0099] Example 2 Pharmacodynamic study of the test drug in the human lung cancer LD1-0025-200662 PDX tumor transplant model 1. Experimental purpose

[0100] An in vivo transplant tumor model was established by subcutaneously inoculating human lung cancer tumor LD1-0025-200662 into 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.

[0101] 2. Experimental Materials

[0102] 2.1 In vivo transplanted tumor model

[0103] LD1-0025-200662 (see Table 8), human lung cancer tumor tissue, passaged to FP3+3 generation, was used for this efficacy test;

[0104] Table 8 Transplanted tumor model

[0105]

[0106] 2.2 Experimental animals

[0107] NU / NU mice, female, age: 42-62 days, weight 18-22 g, were purchased from Zhejiang Weitonglihua Experimental Animal Technology Co., Ltd. (production license: SCXK (Zhejiang) 2019-0001).

[0108] 2.3 Test drugs

[0109] Vehicle: 20% solutol / 80% 100mM sodium acetate buffer pH 5.5;

[0110] ASK-MET: hydrochloride of the compound represented by formula (II), prepared with 20% solutol / 80% 100 mM sodium acetate buffer pH 5.5, concentration: 0.1 mg / mL, 0.3 mg / mL;

[0111] ASK-EGFR: a compound represented by formula (I), prepared with 0.5% sodium carboxymethylcellulose solution (0.5% CMC-Na), concentration: 2 mg / mL;

[0112] RefT: The drug was prepared with 20% solutol / 80% 100 mM sodium acetate buffer pH 5.5, concentration: 0.3 mg / mL; RefT is a small molecule MET kinase inhibitor approved for marketing by USFDA.

[0113] 3. Experimental Methods

[0114] LD1-0025-200662 tumor tissue was inoculated subcutaneously into immunodeficient mice and observed continuously until tumors formed and the tumor volume reached approximately 1000 mm 3 When the tumor tissue was removed, it was cut into pieces of approximately 3 mm × 3 mm × 3 mm (approximately 50 to 90 mg) and inoculated subcutaneously into NU / NU mice. The mice were observed after inoculation and the growth of the tumor was monitored. The average tumor volume of the tumor-bearing mice reached 193.40 mm 3 At the time of random grouping and drug administration, observation was performed. The tumor volume and body weight of all tumor-bearing mice were measured before grouping. The tumor-bearing mice were randomly grouped according to the measured tumor volume. The randomized block design principle was adopted. The mice were first divided into blocks according to the tumor volume, and then the mice in each block were randomly assigned to each treatment group. The day of grouping was day 0. The specific grouping and drug administration information is shown in Table 9 below.

[0115] Table 9 Drug administration and treatment

[0116]

[0117] Note: po: intragastric administration; QD: once a day.

[0118] Dosage volume: The dosage volume was adjusted according to the weight of the tumor-bearing mice (0.1 mL / 10 g).

[0119] 4. Experimental Observation

[0120] Throughout the entire experimental process, the use and observation of experimental animals were carried out in accordance with the relevant regulations of AAALAC Animal Use and Management. After the experimental animals were inoculated with tumor tissue, they were observed every day, and their morbidity and mortality were recorded. In the routine experimental process, all experimental animals were monitored and recorded for behavior, eating, drinking, weight changes, hair gloss and other abnormal conditions.

[0121] 5. Evaluation Metrics

[0122] 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.

[0123] 5.1 Tumor volume and weight of tumor-bearing mice were measured twice a week using a vernier caliper. The tumor volume was calculated using the formula V = 0.5 × a × b 2 , a, b represent the long diameter and wide diameter of the tumor, respectively.

[0124] 5.2 Tumor growth inhibition rate TGI (%) = [1-(Ti-T 0 ) / (Vi-V 0)] × 100, where Ti-T 0 >0; TGI (%) = [1-(Ti-T 0 ) / T 0 ]× 100, of which Ti-T 0 <0. T 0 is the average tumor volume of the drug-treated group at the first dose, Ti is the average tumor volume of the drug-treated group at a certain measurement after the start of drug administration; V 0 is the average tumor volume of the vehicle control group (Vehicle) at the first dose, and Vi is the average tumor volume of the vehicle control group at a certain measurement after the start of drug administration.

[0125] 5.3 Relative tumor proliferation rate T / C (%): The calculation formula is as follows: T / C % = T RTV / C RTV × 100 %(T RTV :RTV in treatment group; C RTV : RTV of negative control group). The relative tumor volume (RTV) was calculated based on the results of tumor measurement. The calculation formula is RTV = Vt / V 0 , where V 0 When the drug is administered in groups (i.e., d 0 ) is the average tumor volume measured, Vt is the average tumor volume at a certain measurement, T RTV With C RTV Get data from the same day.

[0126] 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 Q value > 1 reflects that the combination of drugs has a synergistic effect.

[0127] 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.

[0128] 5.6 The 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 -BW 0 ) / BW 0 ×100, BW i BW is the average body weight at a certain time after the start of drug administration. 0 The average body weight at the time of first administration.

[0129] 6. Statistical analysis

[0130] All data were expressed as Mean ± SEM, and the One-Way ANOVALS D (L) test was used to compare the significant differences in tumor volume and tumor weight among the groups. *P < 0.05 was considered to be significantly different.

[0131] 7. Experimental Results

[0132] 7.1 Tumor volume and weight

[0133] The purpose of this experiment is to establish an in vivo LD1-0025-200662 PDX tumor model by subcutaneously inoculating NU / NU mice with LD1-0025-200662 PDX tumors, and then evaluate the efficacy of the test drug in this tumor model against tumor growth. The changes in tumor volume, tumor images, and statistical analysis results of the treatment group of the test drug acting on the LD1-0025-200662 PDX animal model are shown in Figure 3 , Figure 4 , Table 10, Table 11 and Table 12.

[0134] The drug administration cycle of this efficacy experiment was 21 days, after which the entire efficacy experiment was terminated. _ 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 _ The average tumor volumes of the tumor-bearing mice in the 3mg / kg+ASK-EGFR_20mg / kg and RefT_3mg / kg+ASK-EGFR_20mg / kg groups were 1261.93±184.86mm 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 3Compared with the Vehicle group, the tumor growth inhibition rates TGI (%) of 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 1.96%, 131.27%, 197.93%, 180.72%, 200.00% and 140.79%, respectively. According to the above results, except for the ASK-EGFR_20mg / kg group, all other groups showed statistically significant inhibition of tumor growth (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), showing a significant synergistic effect (Q>1).

[0135] On the day the experiment ended (day 21), the Vehicle group and 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 _ The rates of complete tumor regression in the 3mg / kg+ASK-EGFR_20mg / kg group and the 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 of the tumor-bearing mice were removed and weighed. The results of tumor weight and tumor volume were consistent.

[0136] Table 10 Pharmacodynamics study of the test drugs in the human lung cancer LD1-0025-200662 in vivo transplanted tumor model

[0137]

[0138] 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. a: The proportion of mice with completely disappeared tumors in each treatment group after treatment was observed by naked eye on the 21st day.

[0139] Table 11 Statistical analysis results of the combined drug group compared with the single drug group (Day 21)

[0140]

[0141] 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 ASK-MET, 1 mg / kg group, ▲ p<0.05, p<0.05 was considered to be significantly different; compared with ASK-MET, 3 mg / kg group, ▼ p<0.05, p<0.05 was considered to have significant difference.

[0142] Table 12 Tumor mass weight (g) (Mean ± SEM)

[0143]

[0144] 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.

[0145] 7.2 Changes in mouse body weight

[0146] The results of body weight changes and relative body weight changes of each medication group are shown in Figure 5 and Table 13. During this experiment, mice in all treatment groups did not show significant weight loss and other abnormal symptoms, indicating that tumor-bearing mice had good tolerance to the tested drugs.

[0147] 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

[0148]

[0149] 8. Experimental Summary and Discussion

[0150] 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 a statistically significant inhibitory effect on the growth of subcutaneously transplanted tumors in mice with human lung cancer LD1-0025-200662. Moreover, the ASK-MET_1mg / kg administration group, ASK-MET_3mg / kg administration group, ASK-MET_1mg / kg+ASK-EGFR_20mg / kg administration group and ASK-MET_3mg / kg+ASK-EGFR_20mg / kg administration group were able to cause complete regression of tumors in tumor-bearing mice to varying degrees. At the same time, the anti-tumor efficacy of the ASK-MET_1mg / kg+ASK-EGFR_20mg / kg combined administration group and the ASK-MET_3mg / kg+ASK-EGFR_20mg / kg combined administration group was significantly better than the corresponding single-drug treatment group, showing a significant synergistic effect. In addition, during the entire administration process, there was no obvious abnormality in the weight and behavior of tumor-bearing mice, indicating that tumor-bearing mice had good tolerance to the test drugs.

[0151] Example 3 An open, multicenter, phase I clinical trial of the safety, tolerability, pharmacokinetic characteristics and preliminary efficacy of ASKC202 tablets combined with ASK120067 tablets in patients with advanced solid tumors

[0152] 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.

[0153] Table 14 Trial drugs and medication regimen

[0154]

[0155] The AEs of the 4 subjects recruited in this study in the first treatment cycle were mainly laboratory abnormalities, and significant anti-tumor activity was initially observed in 2 subjects in the first tumor assessment. Therefore, the combination of ASKC202 tablets and ASK120067 tablets in the treatment of locally advanced or metastatic non-small cell lung cancer has broad prospects.

[0156] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference separately. In addition, it should be understood that after reading the above content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

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.

Images