TW202114670A - A use of a combination of an ezh2 inhibitor and a cdk4/6 inhibitor in preparation of medicine for treating tumors - Google Patents

Abstract

The present invention relates to a use of a combination of an EZH2 inhibitor and a CDK4/6 inhibitor in preparation of medicine for treating tumors. Specifically, the EZH2 inhibitor related to the present invention is selected from a compound shown in a formula (I) or pharmaceutically acceptable salts thereof, and the related CDK4/6 inhibitor is selected from a compound shown in a formula (II) or pharmaceutically acceptable salts thereof. The combination of the EZH2 inhibitor and the CDK4/6 inhibitor further enhances anti-tumor effects compared with single drug.

Description

Use of a combination of EZH2 inhibitor and CDK4/6 inhibitor in the preparation of tumor treatment drugs

This application claims the priority of the Chinese patent application CN201910939509.5 whose filing date is September 30, 2019. This application quotes the full text of the aforementioned Chinese patent application.

The invention relates to the use of a combination of an EZH2 inhibitor and a CDK4/6 inhibitor in the preparation of drugs for treating tumors, and belongs to the field of pharmacy.

Breast cancer is one of the most common malignant tumors in women. There are approximately 1.3 million new cases worldwide each year. In my country, the incidence of breast cancer accounts for 7%-10% of the incidence of various malignant tumors throughout the body, and about 18% of all female tumors. At present, the number of patients in China has exceeded 500,000, and its incidence is increasing rapidly. It ranks first in the incidence spectrum of female tumors, and nearly 50% of patients have recurrence and metastasis after treatment. In recent years, with the deepening of tumor molecular biology research, molecular targeted therapy has become more and more widely used in the treatment of breast cancer and has achieved more significant curative effects. It has become the next three traditional modes of surgery, radiotherapy and chemotherapy. The new treatment model is also a hot spot in the field of breast cancer treatment.

The histone methyltransferase encoded by the EZH2 gene is the catalytic component of the polycomb inhibitory complex 2 (PRC2). Compared with normal tissues, the level of EZH2 is abnormally increased in cancer tissues, and the level of EZH2 is the highest in advanced cancer or poor prognosis. In some cancer types, the overexpression of EZH2 occurs at the same time as the amplification of the EZH2 gene. A large number of si/shRNA experimental studies have found that reducing the expression of EZH2 in tumor cell lines can inhibit tumor cell proliferation, migration and invasion or angiogenesis, and lead to cell apoptosis.

At present, EZH2 inhibitors have entered the clinical development stage. The following is a brief list. Tazemetostat (EPZ-6438) developed by Eisai is used for the treatment of non-Hodgkin’s B-cell lymphoma. It is currently in clinical phase II and developed by Constellation. CPI-1205 is used for the treatment of B-cell lymphoma and is currently in clinical phase I. GSK-2816126 developed by GlaxoSmithKline is used for the treatment of diffuse large B-cell lymphoma and follicular lymphoma and is currently in clinical phase I stage.

。WO2017084494A provides an EZH2 inhibitor, the structure is as follows:

.

Cyclin-dependent kinase (Cyclin-dependent kinase, CDK) is a type of serine/threonine kinase that forms a dimer with the corresponding cyclin (Cyclin) to phosphorylate downstream protein molecules, thereby promoting cells The orderly progress of each phase of the cycle realizes cell growth and proliferation. At present, a variety of CDK4/6 selective inhibitors have been in clinical trials or have been approved for listing in foreign countries, including Pfizer’s Palbociclib, Novartis’s Ribociclib and Eli Lilly’s Abemaciclib.

(II)， PCT申請WO2016124067A公開了上述式(II)所示化合物的羥乙基磺酸鹽及其製備方法。WO2014183520 discloses a chemical named 6-acetyl-8-cyclopentyl-5-methyl-2-((5-(piperidin-4-yl)pyridin-2-yl)amino)pyrido[ 2,3-d]pyrimidin-7(8H)-one, the structural formula is as the CDK4/6 inhibitor shown in formula (II), it has significant CDK4/6 inhibitory activity and high selectivity.

(II), PCT application WO2016124067A discloses the isethionate of the compound represented by the above formula (II) and a preparation method thereof.

Xuejiao Song, et al (scientific reports 2016) disclosed that a small molecule EZH2 inhibitor ZLD1039 can selectively block H3K27 methylation and produce anti-tumor effects in breast cancer.

CN109937041A discloses the use of a combination of an EZH2 inhibitor and a BTK kinase inhibitor in the preparation of a medicine for the treatment of cancer, wherein the combination optionally includes a third component, and the third component may be a CDK4/6 inhibitor. There is no report on the mechanism or related experiments of the combination of CDK4/6 inhibitor and EZH2 inhibitor.

The invention provides a use of a combination of an EZH2 inhibitor and a CDK4/6 inhibitor in the preparation of drugs for treating tumors.

In certain embodiments, the EZH2 inhibitor described in the present invention can be selected from CPI-0209, CPI-1205, GSK126, valemetostat, tazemetostat, PF-06821497, DS-3201 GSK-2816126, 3-deazaneplanocin A, HKMT- At least one of I-005 and KM-301.

。In some embodiments, the EZH2 inhibitor in the present invention is a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,

.

The pharmaceutically acceptable salt of the compound represented by formula (I) of the present invention can be hydrochloride, phosphate, hydrogen phosphate, sulfate, hydrogen sulfate, sulfite, acetate, oxalate, malonate, pentane Acid salt, glutamate, oleate, palmitate, stearate, laurate, borate, p-toluenesulfonate, methanesulfonate, isethionate, maleate Acid salt, malate, tartrate, benzoate, pamoate, salicylate, vanillate, mandelate, succinate, gluconate, lactobionate or lauryl sulfonic acid Salt etc.

In some embodiments, the administration dose of EZH2 is selected from 1 to 1600 mg, for example: 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 105 mg, 110 mg, 115 mg, 120 mg, 125 mg, 130 mg, 135 mg , 140 mg, 145 mg, 150 mg, 155 mg, 160 mg, 165 mg, 170 mg, 175 mg, 180 mg, 185 mg, 190 mg, 195 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, 350 mg, 360 mg, 370 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg, 450 mg, 460 mg, 470 mg, 480 mg, 490 mg, 500 mg, 510 mg, 520 mg, 530 mg, 540 mg, 550 mg, 560 mg, 570 mg , 580 mg, 590 mg, 600 mg, 625 mg, 650 mg, 675 mg, 700 mg, 725 mg, 750 mg, 775 mg, 800 mg, 825 mg, 850 mg, 875 mg, 900 mg, 925 mg, 950 mg, 975 mg, 1000 mg, 1025 mg, 1050 mg, 1075 mg, 1100 mg, 1125 mg, 1150 mg, 1175 mg, 1200 mg, 1225 mg, 1250 mg, 1275 mg, 1300 mg, 1325 mg, 1350 mg, 1375 mg, 1400 mg, 1425 mg, 1450 mg, 1475 mg, 1500 mg, 1525 mg, 1550 mg, 1575 mg, 1600 mg, the frequency of administration is twice a day or once a day.

In some embodiments, the dosage of EZH2 is selected from 1-800 mg, and the frequency of administration is twice a day or once a day.

In some embodiments, the administration dose of EZH2 is selected from 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, the frequency of administration is twice a day or once a day.

In some embodiments, the administration dose of EZH2 is selected from 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, the frequency of administration is twice a day.

In some embodiments, the dosage of EZH2 is selected from 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, and the frequency of administration is twice a day or once a day.

In some embodiments, the dosage of EZH2 is selected from 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, and the frequency of administration is twice a day.

In an alternative embodiment, the CDK4/6 inhibitor in the present invention is selected from abemaciclib, ribociclib, palbociclib, alvocidib, trilaciclib, voruciclib, AT-7519, G1T-38, FLX-925, INOC-005, G1T28-1 , BPI-1178, gossypin, G1T30-1, GZ-38-1, P-276-00, staurosporine, R-547, PAN-1215, PD-0183812, AG-024322, NSC-625987, CGP-82996, PD -At least one of 171851.

。In an alternative embodiment, the CDK4/6 inhibitor in the present invention is a compound represented by formula (II) or a pharmaceutically acceptable salt thereof,

.

In the present invention, the pharmaceutically acceptable salt of the compound represented by formula (II) is selected from hydrochloride, phosphate, hydrogen phosphate, sulfate, hydrogen sulfate, sulfite, acetate, oxalate, and malonic acid Salt, valerate, glutamine, oleate, palmitate, stearate, laurate, borate, p-toluenesulfonate, methanesulfonate, isethionate, cis Butenediolate, malate, tartrate, benzoate, pamoate, salicylate, vanillate, mandelic acid, succinate, gluconate, lactobionate, or laurel Sulfonate.

In an alternative embodiment, the CDK4/6 inhibitor is isethionate of the compound represented by formula (II).

In an alternative embodiment, the dosage of the CDK4/6 inhibitor is selected from 1-500 mg, preferably 25-200 mg, more preferably 100-175 mg, and the frequency of administration is once a day or once a day. Twice a day; the dose of specific CDK4/6 inhibitors can be 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 105 mg, 110 mg, 115 mg, 120 mg, 125 mg, 130 mg, 135 mg, 140 mg, 145 mg, 150 mg, 155 mg, 160 mg , 165 mg, 170 mg, 175 mg, 180 mg, 185 mg, 190 mg, 195 mg, 200 mg.

In an optional embodiment, the administration dose of the CDK4/6 inhibitor is selected from 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, and the administration frequency is once a day or once a day. Twice a day.

In an optional embodiment, the dosage of the CDK4/6 inhibitor is selected from 75 mg, 100 mg, 125 mg, or 150 mg, and the frequency of administration is once a day.

In an alternative embodiment, the EZH2 inhibitor is a compound represented by formula (I) or a pharmaceutically acceptable salt thereof, and the CDK4/6 inhibitor is a compound represented by formula (II) or a pharmaceutically acceptable salt thereof.

In an alternative embodiment, the EZH2 inhibitor is a compound represented by formula (I) or a pharmaceutically acceptable salt thereof, and the CDK4/6 inhibitor is a compound represented by formula (II) isethionate.

The tumor in the present invention is selected from breast cancer, ovarian cancer, endometrial cancer, kidney cancer, uterine cancer, prostate cancer, lung cancer, rectal cancer, non-small cell lung cancer, melanoma, pancreatic cancer, sarcoma, osteosarcoma, scleroderma Fibroma, adenoid cystic carcinoma, medulloblastoma, colorectal cancer, thyroid cancer, esophageal cancer, head and neck cancer, urethral cancer, hepatocellular carcinoma, neuroendocrine tumor, glioblastoma, cholangiocarcinoma, testicular tumor, nerve Blastoma, liposarcoma, brain tumor, ureteral tumor, bladder tumor, gallbladder cancer, thyroid tumor, lymphoma, leukemia, rhabdoid tumor, synovial sarcoma, mesothelioma, cervical cancer, colon cancer, gastric cancer, brain cancer , Skin cancer, oral cancer, bone cancer, bladder cancer, fallopian tube tumor, peritoneal tumor, glioma, glioblastoma, head and neck tumor, seminoma, myeloma, chorioepithelial carcinoma, polycythemia vera , Multiple myeloma, the leukemia can be selected from acute bone random leukemia, chronic bone random leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, etc.; the lymphoma is preferably non-Hodgkin's Lymphoma, diffuse large B-cell lymphoma or follicular lymphoma, etc.; the head and neck tumors include maxillary cancer, laryngeal cancer, pharyngeal cancer, tongue cancer, intraoral cancer and the like.

In an alternative embodiment, the tumor in the use provided by the present invention is breast cancer.

In an alternative embodiment, the use provided in the present invention optionally further comprises other anti-tumor agents. When the use provided in the present invention contains other anti-tumor agents, the anti-tumor agent is not a BTK kinase inhibitor.

The method of the combination of the present invention includes, but is not limited to, oral administration, parenteral administration, and transdermal administration. The parenteral administration includes, but is not limited to, intravenous injection, subcutaneous injection, and intramuscular injection.

The present invention provides a method for treating tumors, which comprises administering to a patient a therapeutically effective amount of the above-mentioned EZH2 inhibitor and CDK4/6 inhibitor.

Another aspect of the present invention provides an EZH2 inhibitor for the treatment of tumors, which is used in combination with a CDK4/6 inhibitor.

Another aspect of the present invention provides a CDK4/6 inhibitor for the treatment of tumors, which is used in combination with an EZH2 inhibitor.

In an alternative embodiment, in the method for treating tumors provided by the present invention, the patient is a human.

The present invention provides a pharmaceutical composition comprising the above-mentioned EZH2 inhibitor and CDK4/6 inhibitor and one or more pharmaceutically acceptable excipients, diluents or carriers.

The "combination" in the present invention is a mode of administration, which means that at least one dose of EZH2 inhibitor and CDK4/6 inhibitor is administered within a certain period of time, and both drugs show pharmacological effects. The time limit may be within one administration cycle, for example, within 4 weeks, within 3 weeks, within 2 weeks, within 1 week, or within 24 hours. EZH2 inhibitors and CDK4/6 inhibitors can be administered simultaneously or in no particular order. This period includes treatments in which the EZH2 inhibitor and the CDK4/6 inhibitor are administered by the same route of administration or different routes of administration.

The following examples are used to further describe the present invention, but these examples do not limit the scope of the present invention.

[Example 1. Evaluation of CDK4/6 inhibitor (compound isethionate represented by formula (II)) and EZH2 inhibitor (compound represented by formula (I)) in xxT47D human breast cancer cell subcutaneous xenograft tumor BALB/ c In vivo pharmacodynamics on nude mouse model] 1. Experimental materials 1) Experimental animals Species: mouse; strain: BALB/c nude mice; age and weight: 6-8 weeks old, weighing 18-22 grams; gender: female; number: 32 (excluding the remaining mice in the group); supplier : Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.

2) Experimental drugs CDK4/6 inhibitor: isethionate (drug A) of the compound represented by formula (II); EZH2 inhibitor: a compound represented by formula (I) (drug B).

2. Experimental methods and steps 1) Establishment of xxT47D breast cancer model xxT47D tumor cells were established by isolating xenograft tumor cells constructed from parental T47D tumor cells to establish cell lines in vitro, and the same process was performed twice to complete the establishment. xxT47D tumor cells were cultured adherently in vitro. The culture conditions were RPMI 1640 medium with 10% fetal calf serum, 100 U/ml penicillin and 100 μg/ml streptomycin, and cultured _{at 37°C with 5% CO 2.} Use pancreatin-EDTA for routine digestion and subculture twice a week. When the cell saturation is 80%-90%, the cells are collected, counted, and seeded.

2) Tumor cell inoculation: Estrogen tablets (0.18 mg/tablet) were subcutaneously inoculated on the left back of each mouse. Three days later, 0.2 mL (10×106 cells + Matrigel, volume ratio 1:1) xxT47D cells The mice were inoculated subcutaneously on the right back of each mouse. When the average tumor volume reached 185 mm ³ , the drugs were administered in groups according to the experimental design (Table 1).

Table 1. Grouping of experimental animals and dosing schedule Group N ¹ Compound therapy Dose (mg/kg) Dosing volume (µl/g) ² Route of administration Dosing frequency 1 8 Vehicle A +Vehicle B - 10 po QD3+BID3 2 8 Drug A 25 10 po QD×4W 3 8 Drug B 100 10 po BID×4W 4 8 Drug A 25 10 po QD×4W Drug B 100 10 po BID×4W Note: 1. N: the number of mice in each group; 2. Dosing volume: 10 µl/g based on mouse body weight. If the weight loss exceeds 15%, the dosing regimen should be adjusted accordingly; QD: once a day; BID: twice a day at an 8-hour interval.

3) Test drug configuration table 2. Test substance preparation method Compound Packaging or starting concentration Preparation method Concentration (mg/ml) Storage conditions Vehicle A - Weigh 480 mg of citric acid solids and dissolve them with 50 mL 0.5% CMC-Na solution to a clear and transparent solution. Adjust the pH to 5.0 with 5 M NaOH solution. This solution is called VA. Add 250 μL of Tween- to VA. 80, stir and mix well - 4℃ Vehicle B - Weigh 0.5 g of CMC-Na and dissolve it in distilled water at 60°C until it is completely dissolved, then pipette 1 ml of Tween-80 and add 99 ml of 0.5% CMC-Na to prepare a 1% Tween-80-CMC-Na solution - 4℃ Drug A 500 mg/vial Weigh 36 mg of drug A, grind it in a mortar, add 72 μL of 100% Tween-80 and a small amount of VA solution, grind into a liquid with better fluidity, transfer to a beaker, add VA solution to a final volume of 14.4 mL 2.5 4℃ Drug B 6000 mg/vial Weigh 288 mg of drug B, add 28.8 mL of Vehicle B solution, vortex and sonicate thoroughly to form a 10 mg/ml drug B suspension 10.0 4℃ .

4) Experimental observation, data collection and statistical analysis The formulation of this experimental protocol and any modification have passed the evaluation and approval of the Laboratory Animal Management and Use Committee (IACUC) of Suzhou WuXi AppTec New Drug Development Co., Ltd. The use and welfare of laboratory animals comply with the regulations of the International Laboratory Animal Evaluation and Accreditation Committee (AAALAC). Monitor the animal’s health and death every day. Routine inspections include observation of the effects of tumor growth and drug treatment on the animal’s daily behavior, such as behavioral activities, food and water intake (visual inspection only), and weight changes (measurement of body weight twice a week), Physical signs or other abnormalities. Based on the number of animals in each group, the number of animal deaths and side effects in the group were recorded.

The experimental index is to investigate whether the tumor growth is inhibited, delayed or cured. The tumor diameter was measured with vernier calipers twice a week. The calculation formula of tumor volume is: V=0.5 a × b ² , where a and b represent the long diameter and short diameter of the tumor, respectively.

The anti-tumor efficacy of the compound was evaluated by TGI (%) or the relative tumor proliferation rate T/C (%). Relative tumor proliferation rate T/C (%)=T _TV /C _TV ×100% (T _TV : average TV in the treatment group; C _TV : average TV in the negative control group).

TGI (%), reflects the tumor growth inhibition rate. Calculation of TGI (%): TGI (%)=[(1-(Average tumor volume at the end of a certain treatment group-average tumor volume at the beginning of the treatment group))/(Average tumor at the end of treatment in the solvent control group Volume-average tumor volume at the start of treatment in the solvent control group)]×100%.

After the experiment is over, the weight of the tumor will be detected, and the percentage of _{T weight} /C _{weight will be calculated. T weight} and C _weight represent the tumor weight of the administration group and the vehicle control group, respectively.

Statistical analysis, including the mean and standard error (SEM) of the tumor volume at each time point in each group. The treatment group showed the best treatment effect on the 27th day after the administration at the end of the trial, so based on this data, statistical analysis was performed to evaluate the difference between the groups. The comparison between the two groups was analyzed by T-test. SPSS 17.0 was used for all data analysis. p<0.05 considered a significant difference.

3. Experimental results 1) Mortality, morbidity and weight changes The body weight of experimental animals is used as a reference index for indirect determination of drug toxicity. In this model, none of the administration groups showed significant weight loss (Figure 1). No morbidity. The effect of test substance treatment on the body weight of xxT47D tumor mice is shown in Figure 1 and Figure 2.

2) Tumor volume The tumor volume changes of each group after treatment with the test substance of xxT47D tumor mice are shown in Table 3, and the growth curve of each group of tumors is shown in Figure 3.

Table 3. Tumor volume at different time points in each group Group Tumor volume (mm ³ ) ^a Group 1 Group 2 Group 3 Group 4 0 ^b 185±8 185±7 185±9 185±9 4 230±5 163±7 160±5 169±13 7 274±9 191±7 173±6 147±8 11 301±18 223±11 186±12 135±9 14 407±45 274±25 240±19 123±6 18 495±68 300±24 290±21 123±8 twenty one 565±83 322±31 357±33 145±9 25 617±104 344±34 418±41 159±10 27 714±140 344±40 461±50 157±12 Note: a. Mean ± SEM; b. Days after administration.

3) Anti-tumor efficacy evaluation index Table 4. Anti-tumor efficacy evaluation of drug A and drug B on xxT47D xenograft tumor model (calculated based on tumor volume on the 27th day after administration) treatment Tumor volume (mm ³ ) ^a day 27 T/C ^b (%) TGI ^b (%) p value ^c p value VS group 2 p value VS group 3 Group 1 714±140 - - - Group 2 344±40 48 70 0.024 Group 3 461±50 65 48 0.112 Group 4 157±12 twenty two 105 0.005 0.002 ＜0.001 Note: a. Mean±SEM; b. Tumor growth inhibition is determined by T/C (T/C (%)=T ₂₇ /V ₂₇ ×100%) and TGI (TGI (%)=[1-(T _27- T ₀ )/(V ₂₇ -V ₀ )]×100) calculation; c. p value is calculated based on tumor volume.

Table 5. Tumor weight analysis in each group treatment Tumor weight (g) ^a day 27 T/C ^b (%) p value ^c p value VS group 2 p value VS group 3 Group 1 0.747±0.168 - - Group 2 0.370±0.041 50 0.046 Group 3 0.513±0.056 69 0.205 Group 4 0.183±0.014 twenty four 0.005 0.002 ＜0.001 Note: a. Mean ± SEM; b. Tumor growth inhibition is _{calculated by T weight} / C _weight = TW _treatment / TW _vehicle ; c. P value is calculated based on tumor weight.

27 days after the start of administration, the average tumor volume of tumor mice in the solvent control group reached 714 mm ³ . The average tumor volume of the test substance 25 mg/kg drug A and its combination with 100 mg/kg drug B were 344 mm ³ and 157 mm ^{3 respectively} . Compared with the solvent control group, both showed significant inhibition. Tumor effects (p values of 0.024 and 0.005, respectively), the combination group showed stronger anti-tumor activity compared with drug A and drug B single-drug groups, and the difference was significant, p values were 0.002 and <0.001, respectively. The average tumor volume of the test substance 100 mg/kg drug B single agent was 461 mm ³ , which did not show significant tumor suppressor effect compared with the solvent control group (p value is 0.112).

The result of tumor weight is basically consistent with the result of tumor volume.

In summary, drug A single agent showed significant anti-tumor activity on the xxT47D human breast cancer xenograft tumor model at the dose of the test protocol. Compared with a single drug, the combined application of drug A and drug B can further enhance the anti-tumor effect.

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Figure 1. The body weight change curve of tumor mice in different groups. Figure 2. The relative body weight change curve of tumor mice in different groups. Figure 3. Tumor growth curve.

Claims (10)

The use of an EZH2 inhibitor combined with a CDK4/6 inhibitor in the preparation of a tumor medicine. The use according to claim 1, wherein the EZH2 inhibitor is selected from CPI-0209, CPI-1205, GSK126, valemetostat, tazemetostat, PF-06821497, DS-3201 GSK-2816126, 3-deazaneplanocin A, HKMT-I -005, KM-301 or a compound represented by formula (I) or a pharmaceutically acceptable salt thereof, preferably a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,

. The use according to claim 1, wherein the CDK4/6 inhibitor is selected from abemaciclib, ribociclib, palbociclib, alvocidib, trilaciclib, voruciclib, AT-7519, G1T-38, FLX-925, INOC-005, G1T28-1 , BPI-1178, gossypin, G1T30-1, GZ-38-1, P-276-00, staurosporine, R-547, PAN-1215, PD-0183812, AG-024322, NSC-625987, CGP-82996, PD -171851 or at least one of the compound represented by formula (II) or its pharmaceutically acceptable salt, preferably the compound represented by formula (II) or its pharmaceutically acceptable salt,

. The use according to claim 3, wherein the CDK4/6 inhibitor is an isethionate of the compound represented by formula (II). The use according to any one of claims 1-4, wherein the tumor is selected from breast cancer, ovarian cancer, endometrial cancer, kidney cancer, uterine cancer, prostate cancer, lung cancer, rectal cancer, non-small cell lung cancer, black Tumor, pancreatic cancer, sarcoma, osteosarcoma, desmoid tumor, adenoid cystic carcinoma, medulloblastoma, colorectal cancer, thyroid cancer, esophageal cancer, head and neck cancer, urethral cancer, hepatocellular carcinoma, neuroendocrine tumor, Glioblastoma, cholangiocarcinoma, testicular tumor, neuroblastoma, liposarcoma, brain tumor, ureteral tumor, bladder tumor, gallbladder cancer, thyroid tumor, lymphoma, leukemia, rhabdoid tumor, synovial sarcoma, mesothelioma , Cervical cancer, colon cancer, stomach cancer, brain cancer, skin cancer, oral cancer, bone cancer, bladder cancer, fallopian tube tumor, peritoneal tumor, glioma, glioblastoma, head and neck tumor, seminoma, At least one of myeloma, chorioepithelial carcinoma, polycythemia vera, and multiple myeloma, preferably breast cancer. The use according to claim 5, wherein the dose of the EZH2 inhibitor is selected from 1 to 1600 mg, and the frequency of administration is twice a day or once a day. The use according to claim 6, wherein the dose of the EZH2 inhibitor is selected from 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg or 450 mg, and the frequency of administration is twice a day. The use according to claim 5, wherein the dose of the CDK4/6 inhibitor is selected from 1-500 mg, and the frequency of administration is once a day or twice a day. The use according to claim 8, wherein the dose of the CDK4/6 inhibitor is selected from 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg or 175 mg, and the frequency of administration is once a day Or twice a day. A pharmaceutical composition comprising the EZH2 inhibitor and CDK4/6 inhibitor according to any one of claims 1-9, and one or more pharmaceutically acceptable excipients, diluents or carriers.

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