CN113840608A - Application of CDK4/6 inhibitor and VEGFR inhibitor in preparation of drugs for treating tumors - Google Patents

Abstract

Use of a CDK4/6 inhibitor in combination with a VEGFR inhibitor in the preparation of a medicament for the treatment of a neoplastic disease. Specifically, the CDK4/6 inhibitor is a compound shown as a formula (I) or a pharmaceutically acceptable salt thereof, and the VEGFR inhibitor is apatinib or a pharmaceutically acceptable salt thereof.

Description

Application of CDK4/6 inhibitor and VEGFR inhibitor in preparation of drugs for treating tumors

The present application claims priority from chinese patent application CN201910460310.4 filed on 30/5/2019. The present application refers to the above-mentioned chinese patent application in its entirety.

Technical Field

The application relates to application of a CDK4/6 inhibitor and a VEGFR inhibitor in preparation of a medicine for treating tumor diseases.

Background

Breast cancer is one of the most common malignancies in women, with about 130 million cases worldwide each year. In China, the incidence rate of breast cancer accounts for 7% -10% of the incidence rate of various malignant tumors of the whole body, approximately accounts for 18% of all female tumors, the number of patients in China currently exceeds 50 ten thousand, the incidence rate is rapidly increased, the first female tumor incidence spectrum is listed in some big cities, and nearly 50% of patients relapse and transfer after treatment. In recent years, with the deepening of the research of tumor molecular biology, molecular targeted therapy is more and more widely applied to breast cancer treatment and obtains more remarkable curative effect, becomes a brand new treatment mode following three traditional modes of operation, radiotherapy and chemotherapy, and is also a hotspot of the research in the field of breast cancer treatment at present.

Cyclin-dependent kinases (CDKs) are serine/threonine kinases, which form dimers with corresponding cyclins to phosphorylate downstream protein molecules, thereby promoting the orderly progression of each phase of the cell cycle and realizing cell growth and proliferation. Currently, a variety of CDK4/6 selective inhibitors are available in clinical trials or approved in the market, including Palbociclib from Perey, Ribociclib from Nowa, Abemaciclib from Paris, and the like.

WO2014183520 discloses a CDK4/6 inhibitor with a chemical name of 6-acetyl-8-cyclopentyl-5-methyl-2- ((5- (piperidin-4-yl) pyridin-2-yl) amino) pyrido [2,3-d ] pyrimidin-7 (8H) -one and a structural formula shown as a formula (I), and the CDK4/6 inhibitor has obvious CDK4/6 inhibitory activity and high selectivity,

WO2016124067A discloses isethionate salts of the compounds of formula (I) above and a process for their preparation.

Much research has been carried out on the combination of CDK4/6 inhibitors with other drugs, WO2017193141A discloses that CDK4/6 inhibitors in combination with EGFR inhibitors can be used to treat triple negative breast cancer; WO2016024-232A discloses methods of treating cancer with CDK4/6 inhibitors in combination with BTK kinase inhibitors; lori S.Hart et al found that CDK4/6 inhibitors and MEK inhibitors had synergistic effects in Preclinical Neuroblastoma "clinical Therapeutic Synergy of MEK1/2 and CDK4/6 Inhibition in Neuroblastoma"; CN103781480A discloses that CDK4/6 inhibitors in combination with FGFR kinase inhibitors are useful for the treatment of cancer; bollard et al, "Palbociclib (PD-0332991) a selective CDK4/6 inhibitor, restricts tumourgrowth in clinical models of hepatocellular carcinosa" disclose that Palbociclib alone or in combination with sorafenib may become a new strategy for treating hepatocellular carcinoma; CN106029097A discloses abemaciciclib and ramucirumab used in combination for treating non-small cell lung cancer; CN108883182A discloses the combination of abemaciciclib and ramucirumab for the treatment of mantle cell lymphoma.

There is currently little research on the use of CDK4/6 inhibitors in combination with small-molecule VEGFR inhibitors for the treatment of cancer.

Disclosure of Invention

The application provides application of a CDK4/6 inhibitor and a VEGFR inhibitor in preparation of a medicine for treating tumor diseases.

The CDK4/6 inhibitor described herein may be selected from compounds of formula (I) or a pharmaceutically acceptable salt thereof,

in the present application, the pharmaceutically acceptable salt of the compound represented by formula (I) is selected from hydrochloride, phosphate, hydrogen phosphate, sulfate, hydrogen sulfate, sulfite, acetate, oxalate, malonate, valerate, glutamate, oleate, palmitate, stearate, laurate, borate, p-toluenesulfonate, methanesulfonate, isethionate, maleate, malate, tartrate, benzoate, pamoate, salicylate, vanillite, mandelate, succinate, gluconate, lactobionate or laurylsulfonate, preferably isethionate, and has a structure represented by formula (II),

in alternative embodiments, the VEGFR inhibitor is selected from the group consisting of VEGFR-2 inhibitors, preferably the VEGFR-2 inhibitor is apatinib or a pharmaceutically acceptable salt thereof.

In some embodiments, the pharmaceutically acceptable salt of apatinib is selected from the group consisting of mesylate, maleate, tartrate, succinate, acetate, difluoroacetate, fumarate, citrate, benzenesulfonate, benzoate, naphthalenesulfonate, lactate, malate, hydrochloride, hydrobromide, sulfate, and phosphate, and in preferred embodiments, the pharmaceutically acceptable salt of apatinib is mesylate.

In alternative embodiments, the VEGFR inhibitor is apatinib mesylate.

The neoplastic disease described herein is selected from the group consisting of sarcoma, lymphoma, lung cancer, bronchial cancer, prostate cancer, pancreatic cancer, gastrointestinal cancer, colon cancer, rectal cancer, colorectal adenoma, thyroid cancer, liver cancer, intrahepatic bile duct cancer, hepatocellular cancer, adrenal cancer, gastric tumor, glioma, glioblastoma, endometrial cancer, melanoma, kidney cancer, renal pelvis cancer, bladder cancer, uterine corpus cancer, cervical cancer, vaginal cancer, ovarian cancer, multiple myeloma, esophageal cancer, leukemia, acute myelogenous leukemia, chronic myelogenous leukemia, lymphocytic leukemia, myelogenous leukemia, brain tumor, brain cancer, oral and pharyngeal cancer, laryngeal cancer, small intestine cancer, non-hodgkin's lymphoma, melanoma, villous colon adenoma, neoplasm, epithelial cancer, breast cancer, basal cell carcinoma, squamous cell carcinoma, actinic keratosis, carcinoma, cervical cancer, colon carcinoma, cervical cancer, renal carcinoma, bladder cancer, uterine carcinoma, cervical carcinoma, cervical carcinoma, tumor diseases, neck or head tumors, essential thrombocythemia, myeloid metaplasia myelofibrosis and macroglobulinemia, preferably breast cancer, hepatocellular carcinoma, colon cancer.

In alternative embodiments, the breast cancer is hormone receptor positive breast cancer.

In alternative embodiments, the breast cancer is a triple negative breast cancer.

In alternative embodiments, the breast cancer is her2 positive breast cancer.

In alternative embodiments, the CDK4/6 inhibitor is administered in an amount selected from the group consisting of 1-500mg, preferably 50-200mg, more preferably 100-150mg, once daily, twice daily, preferably once daily.

In alternative embodiments, the CDK4/6 inhibitor is administered in a dose of 50mg, 75mg, 100mg, 125mg, 150mg, 175mg, preferably 100mg, 125mg or 150mg, once daily, twice daily, preferably once daily.

In alternative embodiments, the VEGFR inhibitor is administered at a dose of 100-500mg once a day, once in two days, once in three days, two days off five days off, seven days off.

In alternative embodiments, the dose of the VEGFR inhibitor is 200mg, 225mg, 250mg, 275mg, 300mg, 325mg, 350mg, 375mg, 500mg, preferably 250mg or 375mg, administered once a day, five days off for two days, seven days off for seven days.

In alternative embodiments, the CDK4/6 inhibitor is administered in an amount selected from 100mg, 125mg, or 150mg once daily, the VEGFR inhibitor is administered in an amount of 250mg or 375mg once daily, five days off for two days, seven days off for seven days.

In alternative embodiments, the CDK4/6 inhibitor is administered at a dose of 100mg once a day, the VEGFR inhibitor is administered at a dose of 250mg or 375mg once a day, five days off for two days, seven days off for seven days.

In alternative embodiments, the CDK4/6 inhibitor is administered in a dose of 125mg once a day frequently, and the VEGFR inhibitor is administered in a dose of 250mg or 375mg once a day frequently, two days off five days off seven days on seven days.

In alternative embodiments, the CDK4/6 inhibitor is administered at a dose of 150mg once a day, the VEGFR inhibitor is administered at a dose of 250mg or 375mg once a day, five days off for two days, seven days off for seven days.

The routes of combination described herein include, but are not limited to, oral, parenteral, transdermal, including, but not limited to, intravenous, subcutaneous, intramuscular.

The present application provides a method of treating a neoplastic disorder comprising administering to a subject a therapeutically effective amount of the CDK4/6 inhibitor and VEGFR inhibitor described above.

The present application provides a pharmaceutical composition comprising an inhibitor of CDK4/6, a VEGFR inhibitor as described above and one or more pharmaceutically acceptable excipients, diluents or carriers.

"combination" as used herein is a mode of administration and refers to the administration of at least one dose of a CDK4/6 inhibitor and a VEGFR inhibitor over a period of time, wherein both drugs exhibit pharmacological effects. The time period may be within one administration cycle, preferably within 4 weeks, within 3 weeks, within 2 weeks, within 1 week, or within 24 hours. The CDK4/6 inhibitor and the VEGFR inhibitor may be administered simultaneously or sequentially. Such terms include treatments wherein the CDK4/6 inhibitor, the VEGFR inhibitor, are administered by the same route of administration or different routes of administration.

Drawings

FIG. 1 weight change in tumor-bearing mice following administration of test subjects in the xxT47D xenograft tumor model;

FIG. 2 relative weight changes in tumor-bearing mice following administration of test subjects in the xxT47D xenograft tumor model;

FIG. 3. growth curves of tumors in each group.

Detailed Description

The present application is further described below with reference to examples, but these examples do not limit the scope of the present application.

EXAMPLE 1 Compound isethionate (drug A) of formula (I) Single drug or in combination with Apatinib mesylate (drug B) in vivo pharmacodynamic study in a BALB/c nude mouse model of xxT47D human breast cancer cells (ER +, HER2-) subcutaneous xenograft tumors

1. Experimental Material

Drug A was prepared using the method disclosed in WO 2016124067A;

mice: strain: BALB/c nude mice; week age and body weight: the weight of the patient is 18-22 g after the patient is 6-8 weeks old; sex: a female; quantity: 42 (excluding the group of remaining mice); the supplier: shanghai Spiral-BiKai laboratory animals Co., Ltd.

Cell source: xxT47D tumor cells cell lines were established by in vitro isolation of xenograft tumors constructed from parental T47D tumor cells (source: ATCC, HTB-133).

2. Experimental methods and procedures

1) Establishment of xxT47D breast cancer model

xxT47D tumor cells by parental T47D tumor cellsThe cell-constructed xenograft tumor is separated in vitro to establish a cell line, and the same process is carried out for 2 times to complete the establishment. xxT47D tumor cells are cultured in vitro adherent culture in RPMI 1640 medium supplemented with 10% fetal bovine serum, 100U/ml penicillin and streptomycin 100. mu.g/ml at 37 deg.C with 5% CO₂And (5) culturing. Passage was performed twice a week with conventional digestion treatment with pancreatin-EDTA. When the saturation degree of the cells is 80% -90%, collecting the cells, counting and inoculating.

2) Tumor cell inoculation

A tablet of estrogen (0.18 mg/tablet) was subcutaneously inoculated into the left back of each mouse, and three days later, 0.2mL (10 × 10e6 cells + Matrigel, volume ratio 1:1) of xxT47D cells were subcutaneously inoculated into the right back of each mouse, and the average tumor volume reached 173mm³The group administration was started according to the experimental design (table 1).

TABLE 1 Experimental animal groups and dosing regimens

Note: n is the number of mice in each group; the administration volume: according to the weight of the mouse, 10. mu.l/g. If the body weight is reduced by more than 15%, the dosage regimen should be adjusted accordingly, (Vehicle A + Vehicle B) as the solvent group.

3) Test drug configuration

TABLE 2 test drug configuration

4) The experimental index is to investigate whether the tumor growth is inhibited, delayed or cured. Tumor diameters were measured twice weekly using a vernier caliper. The formula for tumor volume is: v is 0.5a × b²And a and b represent the major and minor diameters of the tumor, respectively.

The tumor suppressor therapeutic effect of the compound was evaluated as TGI (%) or relative tumor proliferation rate T/C (%). TGI (%), reflecting the rate of tumor growth inhibition. Calculation of TGI (%): TGI (%) ═ 1 (average tumor volume at the end of administration in a certain treatment group-average tumor volume at the start of administration in the treatment group))/(average tumor volume at the end of treatment in the solvent control group-average tumor volume at the start of treatment in the solvent control group) × 100%.

Tumor weights will be measured at the end of the experiment and the percent T/C weight calculated, Tweight and cwight indicating tumor weights in the dosing and vehicle control groups, respectively.

Statistical analysis, including mean and Standard Error (SEM) of tumor volume for each time point for each group. Treatment groups showed the best treatment effect at day 21 after dosing at the end of the trial, and therefore statistical analysis was performed based on this data to assess differences between groups. The comparisons between two groups were analyzed using T-test, the comparisons between three or more groups were analyzed using one-way ANOVA, and if there was a significant difference in F-value, the measurements were performed using the Games-Howell method. If there is no significant difference in F value, analysis is performed by the Dunnet (2-sized) method. All data analyses were performed with SPSS 17.0. Significant differences were considered with p < 0.05.

3. Results of the experiment

1) Weight change

The effect of subject treatment on body weight of xxT47D tumor-bearing mice is shown in fig. 1 and 2.

2) Change in tumor volume

Changes in tumor volume in groups following treatment with xxT47D tumor-bearing mouse test agent are shown in table 3.

TABLE 3 tumor volumes at different time points of each group

Note: a. mean ± SEM; b. days after administration

3) Tumor growth curve

The growth curves of the tumors in each group are shown in FIG. 3.

4) Evaluation index of antitumor drug efficacy

TABLE 4 evaluation of the tumor-inhibiting efficacy of drug A and drug B on the xxT47D xenograft tumor model (calculated based on tumor volume on day 21 post-administration)

Note: a. mean ± sem.b. tumor growth inhibition by T/C and TGI (%) ═ 1- (T)₂₁-T ₀)/(V ₂₁-V ₀)]X 100). c.p values were calculated from tumor volumes.

TABLE 5 gravimetric tumor analysis of drug A and drug B groups

Note: a. mean. + -. SEM. b. Tumor growth inhibition was calculated from T/C weight-TW treatment/TW control. c.p values were calculated from tumor weights.

4. Discussion of the experiments

21 days after the start of the administration, the average tumor volume of the tumor-bearing mice in the solvent control group reached 1283mm³The mean tumor volumes of the test substance 25mg/kg of the drug A and the test substance 50mg/kg of the drug B are 724mm respectively³And 704mm³Compared with a solvent control group, the compound has obvious tumor inhibition effect (p values are 0.019 and 0.015 respectively).

The mean tumor volume in the 25mg/kg drug A combined with 50mg/kg drug B treatment group was 354mm³The difference was significant (p < 0.001) compared to the solvent group, and compared to the respective monomersCompared with the traditional Chinese medicine group, the combined group shows stronger antitumor activity, and the difference is obvious, and the p values of the combined group of the medicine A and the medicine B are 0.022 and 0.003 respectively compared with the single medicine. Tumor weight results (table 5) are essentially consistent with tumor volume results.

Taken together, drug a and drug B, taken alone, showed significant anti-tumor activity at the dose of the experimental protocol in the xxT47D human breast cancer xenograft tumor model. Compared with single medicine, the combined application of the medicine A and the medicine B can further enhance the anti-tumor effect.

Although specific embodiments of the present invention have been described above, it will be appreciated by those skilled in the art that these are merely illustrative and that various changes or modifications may be made without departing from the principles and spirit of the invention. The scope of the invention is therefore defined by the appended claims.

Claims (12)

1. Use of a CDK4/6 inhibitor in combination with a VEGFR inhibitor for the preparation of a medicament for the treatment of a neoplastic disease.
2. The use according to claim 1, wherein said CDK4/6 inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof,

   
3. the use according to claim 2, wherein the pharmaceutically acceptable salt of the compound of formula (I) is isethionate.
4. The use according to claim 1, wherein the VEGFR inhibitor is selected from VEGFR-2 inhibitors, preferably wherein the VEGFR-2 inhibitor is apatinib or a pharmaceutically acceptable salt thereof.
5. The use of claim 4, wherein the VEGFR inhibitor is apatinib mesylate.
6. The use according to any one of claims 1 to 5, wherein the neoplastic disease is selected from the group consisting of sarcoma, lymphoma, lung cancer, bronchial cancer, prostate cancer, pancreatic cancer, gastrointestinal cancer, colon cancer, rectal cancer, colorectal adenoma, thyroid cancer, liver cancer, intrahepatic bile duct cancer, hepatocellular cancer, adrenal cancer, stomach cancer, gastric tumor, glioma, glioblastoma, endometrial cancer, melanoma, kidney cancer, renal pelvis cancer, bladder cancer, uterine body cancer, cervical cancer, vaginal cancer, ovarian cancer, multiple myeloma, esophageal cancer, leukemia, acute myelogenous leukemia, chronic myelogenous leukemia, lymphocytic leukemia, myelogenous leukemia, brain tumor, brain cancer, oral and pharyngeal cancer, laryngeal cancer, small intestine cancer, non-Hodgkin's lymphoma, melanoma, villous colon adenoma, neoplasm, epithelial cancer, breast cancer, basal cell carcinoma, Squamous cell carcinoma, actinic keratosis, tumor diseases, neck or head tumors, essential thrombocythemia, myeloid metaplasia myelofibrosis and macroglobulinemia, preferably breast cancer, hepatocellular carcinoma, colon cancer.
7. The use of claim 6, wherein the breast cancer is hormone receptor positive breast cancer.
8. Use according to claim 6 wherein the CDK4/6 inhibitor is administered in an amount selected from the group consisting of 1 to 500mg, preferably 50 to 200mg, more preferably 100 to 150mg, once daily, twice daily, preferably once daily.
9. Use according to claim 8 wherein the CDK4/6 inhibitor is administered in a dose of 50mg, 75mg, 100mg, 125mg, 150mg, 175mg, preferably 100mg, 125mg or 150mg, once daily, twice daily, preferably once daily.
10. The use according to any of claims 8-9, wherein the VEGFR inhibitor is administered in a dose of 100-500mg once a day, once in two days, once in three days, five days off for two days, seven days off for seven days.
11. The use of claim 10, wherein the dose of the VEGFR inhibitor is 200mg, 225mg, 250mg, 275mg, 300mg, 325mg, 350mg, 375mg, 500mg, preferably 250mg or 375mg, administered once a day, five days off for two days, seven days off for seven days.
12. A pharmaceutical composition comprising the inhibitor of CDK4/6 and VEGFR inhibitor of any one of claims 1-11 and one or more pharmaceutically acceptable excipients, diluents, or carriers.

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