Use of tyrosine kinase inhibitor

Abstract

Disclosed in the present invention is a combined pharmaceutical composition for treating tumors, comprising a chemotherapeutic drug and at least one compound represented by formula (I). The compound has the effect of enhancing the efficacy of the chemotherapeutic drug, thereby reducing the dose of the chemotherapeutic drug and diminishing its toxic and side effects.

Description

Use of a tyrosine kinase inhibitor TECHNICAL FIELD

[0001] The present application relates to the field of pharmaceutical technology, and in particular to a combination of a compound and a chemotherapeutic drug. BACKGROUND

[0002] The drug treatment of tumors mainly includes: chemotherapy, small molecule targeted therapy, and tumor immunotherapy. Among them, more than half of tumor patients still need to rely on conventional chemotherapeutic drugs. Conventional chemotherapy has high toxicity, and patients often have serious toxic reactions.

[0003] Chemotherapy synergistic and detoxification drugs can effectively expand the clinical use of chemotherapeutic drugs. Developing synergistic and detoxification drugs for chemotherapeutic drugs can further expand the clinical use of chemotherapeutic drugs, and with the maturation of the market penetration of chemotherapeutic drugs, synergistic and detoxification drugs will rapidly expand, have easier market acceptance than targeted drugs and biological therapy drugs, and form industrialization profit more quickly. The drugs currently used in combination with chemotherapy in clinical practice can only alleviate some adverse reactions, and cannot improve the efficacy of chemotherapeutic drugs.

[0004] In theory, a chemical drug sensitizer can increase the biological activity of a chemical drug, which is equivalent to reducing the dosage of the chemical drug. The chemical sensitizer may also increase the biological toxicity of the chemical drug on the basis of increasing the biological activity of the chemical drug. An ideal sensitizer can improve the biological activity of the chemical drug without increasing the adverse reactions of the chemotherapeutic drug.

[0005] CN113336768 A discloses a multi-target tyrosine kinase inhibitor and a preparation method thereof, and the chemical structural formula is shown as (I), SUMMARY

[0006] An object of the present application is to provide a combination drug composition for treating tumors, which comprises a chemotherapeutic drug and at least one compound as shown in formula (I),

[0007] wherein R1 is selected from the following substituents: carboxyl-substituted C3-C8 alkyl acyl, substituted or unsubstituted phosphono,

[0008] R2 is selected from the following groups: hydrogen, halogen, R5-substituted C1-C8 alkyl, R5-substituted C6-C12 aryl;

[0009] R3, R4 are selected from the following groups: hydrogen, R5-substituted C1-C6 alkyl, or R3, R4 form a five- to twelve-membered aliphatic heterocycle;

[0010] R5is selected from the group consisting of hydrogen, 1-3 halogens, hydroxyl, C1-C6alkoxy, C6-C12aryl.

[0011] The compound of formula (I) has been found to have a sensitizing effect on chemotherapy, and is particularly suitable for sensitizing the anti-tumor effect of chemotherapeutic drugs, thereby reducing the dosage of the chemotherapeutic drugs, and in turn reducing the degree of toxicity, the incidence of adverse reactions and the incidence of serious adverse events caused by chemotherapy, so as to achieve better therapeutic effect and quality of life for patients.

[0012] Preferably, the compound of formula (I) is selected from the compounds of Table 1.

[0013] Table 1 Compounds of formula (I)

[0014] Preferably, the chemotherapeutic drug is selected from drugs affecting nucleic acid biosynthesis, drugs affecting DNA structure and function, drugs interfering with the transcription process and preventing RNA synthesis, drugs inhibiting protein synthesis and function, and drugs regulating hormone balance in the body.

[0015] Preferably, the chemotherapeutic drug is selected from alkylating agents, antimetabolites, antitumor antibiotics, platinum drugs and plant-derived antitumor drugs.

[0016] Preferably, the chemotherapeutic drug is at least one of cyclophosphamide, fluorouracil, doxorubicin, daunorubicin, epirubicin, actinomycin, carboplatin, cisplatin, oxaliplatin, vinblastine, camptothecin, paclitaxel and docetaxel.

[0017] Preferably, the chemotherapeutic drug is at least one of oxaliplatin or paclitaxel.

[0018] Preferably, the compound is compound 10.

[0019] Preferably, the combination composition of the present application is an injection, including but not limited to: water injection, lyophilized powder injection, emulsion injection, suspension injection, liposome injection, microsphere injection, nanoparticle injection, depot controlled-release injection, gel-type injection, etc.; in terms of injection devices and packaging, including single / multiple dose needle-free injection, powder syringe, pre-filled injection, powder / liquid pre-mixed syringe, subcutaneous implantable syringe, etc.

[0020] Preferably, the combination composition of the present application further comprises pharmaceutical excipients, including aqueous solvents, non-aqueous solvents, additional agents, additional agents for increasing the solubility of the main drug, additional agents for helping the suspension or emulsification of the main drug, additional agents for preventing oxidation of the main drug, additional agents for adjusting pH, additional agents for inhibiting microbial reproduction, additional agents for reducing pain, etc.

[0021] According to the experimental research of the present application, the therapeutically effective amount of the compound shown as formula (I) in the combined drug composition of the present application in animal (mice) in vivo experiment is 0.01 mg-1.0 mg / kg body weight / time, and the dosage for human body is 0.05 mg-5 mg / 60 kg body weight / time according to the dose conversion of the FDA Guideline for Drug (Estimating the Maximum Safe Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers, 2005, FDA), and the preferred human drug dosage range is 0.5 mg-2.5 mg / 60 kg body weight / time.

[0022] Preferably, the content of the compound shown as formula (I) in the combined drug composition of the present application is 0.05 mg-5 mg, and more preferably 0.5 mg-2.5 mg. The content refers to the total amount of the active ingredient compound in the single use of the drug, and those skilled in the art can determine the unit dosage form according to the content and the needs of use, including but not limited to the content of the compound shown as formula (I) in each bottle, each tablet, each pill, each bottle, each milliliter, each ten milliliter, each vial, etc.

[0023] Another object of the present application is to provide the use of a compound shown as formula (I) in the preparation of a drug for treating tumors, which can enhance the efficacy of the active ingredient. Preferably, the active ingredient of the drug is a chemotherapeutic drug.

[0024] Another object of the present application is also to provide the use of the above combined drug composition in the preparation of a drug for treating tumors. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1: Sensitization effect of the compound on paclitaxel;

[0026] Figure 2: Sensitization effect of the compound on oxaliplatin;

[0027] Figure 3: Sensitization effect of the compound on 5-Fu

[0028] Figure 4: Inhibition effect of the compound on different kinases;

[0029] Figure 5: Synergistic effect of different MET inhibitors combined with paclitaxel;

[0030] Figure 6: Synergistic effect of different TRK inhibitors combined with paclitaxel;

[0031] Figure 7: Sensitization effect of different compounds on paclitaxel (*: P<0.05 vs paclitaxel group);

[0032] Figure 8: Sensitization effect of different doses of compound 10 on paclitaxel. DETAILED DESCRIPTION

[0033] The technical solutions of the present application will be further illustrated below in combination with some embodiments, and the following embodiments do not constitute any limitation to the present application.

[0034] Compounds 1-24 were prepared according to the synthetic method disclosed in Chinese patent CN 113336768 B.

[0035] Other reagent materials of the present application are commercially available products.

[0036] The materials and reagents used in the following embodiments are as follows:

[0037] Material sources:

[0038] Experimental animals: female BALB / c mice, weighing 18-22 grams, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0039] Cells: human gastric cancer SNU-5 cells, human colorectal adenocarcinoma DLD-1 cells, mouse embryonic fibroblast NIH3T3 cells, human colon adenocarcinoma KM12 cell line, and human lung cancer N87 cells were purchased from ATCC.

[0040] Drugs and reagents: fetal bovine serum (FBS), GIBCO, batch number: 1640958, 1966174C; RPMI 1640, GIBCO, batch number: 8118032; DMEM, GIBCO, batch number: 8114057, 8113368; Pen Strep (double antibiotic), GIBCO, batch number: 1582957; Hyclone, batch number: J150001; Paclitaxel injection, Sichuan Huiyu Pharmaceutical Co., Ltd., batch number: 2412300412; Fluorouracil injection (5-Fu), Hainan Zhoutai Pharmaceutical Co., Ltd., batch number: 20191201; Oxaliplatin for injection, Qilu Pharmaceutical Co., Ltd., batch number: 3H0306B02; Apatinib, Jiangsu Hengrui Medicine Co., Ltd., batch number: 220127KF; Carma, Novartis, batch number: 361926; Entrectinib, Shanghai Aladdin Biochemical Technology Co., Ltd., batch number: C2214404.

[0041]

Example 1

[0042] The purpose of this example is to investigate the sensitization effect of compound 10 of formula (I) on the chemotherapeutic drug paclitaxel.

[0043] 1.1 Experimental method

[0044] The required number of female BALB / c mice were housed in the SPF animal room. The temperature in the animal room was 20-25°C, the relative humidity was 40%-70%, the light and dark illumination was 12 hours each, and the animals were free to drink water and eat.

[0045] The human gastric cancer SNU-5 cell line was obtained from the American Type Culture Collection. Under the culture conditions of 5% CO2, 95% air, saturated humidity, and 37°C, the SNU-5 cells were routinely cultured in a culture medium containing 20% fetal bovine serum. The SNU-5 cells in the logarithmic growth phase were collected, resuspended in 50% serum-free medium and 50% Matrigel after cell counting, and the cell concentration was adjusted to 1.0 x 10^7 cells / mL; the cells were placed in an ice box, and the cell suspension was drawn up with a 1 mL syringe and injected subcutaneously into the right axillary of the mouse, with 200 μL (5.0 x 10^6 cells per mouse) per animal to establish a tumor transplantation model.

[0046] When the tumor volume reached about 150 mm 3 The animals were randomly divided into groups according to the tumor volume, with 10 animals per group. The tumor diameter was measured twice a week after the start of the experiment, and the animal body weight was measured and recorded at the same time. After the experiment was completed, the tumor was weighed.

[0047] Drug administration began on the day of grouping and ended after 21 days. The animal grouping and drug administration scheme are shown below. Paclitaxel was administered via the tail vein, once a week for a total of 3 weeks. The other drugs were administered by gavage, with a drug administration volume of 10 mL / kg, once a day for a total of 3 weeks. The drug administration doses are shown in Table 2.

[0048] Table 2

[0049] Apatinib drug preparation: Before administration, an accurately weighed portion of apatinib powder was taken, sterile water for injection was added, vortexed and oscillated, and ultrasonicated for about 10 min to disperse the drug uniformly, obtaining a suitable concentration of the drug preparation, which was prepared on the spot.

[0050] Paclitaxel drug preparation: The preparation was prepared according to the instructions and diluted to the required concentration according to the drug administration requirements.

[0051] Compound 10 drug preparation: The preparation was prepared to the required concentration according to the method disclosed in Chinese Patent CN 113336768 B.

[0052] On the 22nd day, the tumor diameter was measured and the animal body weight was measured, and the animals were euthanized. The tumor mass was stripped and photographed and weighed. The animals were subjected to gross autopsy, and the internal organs were observed macroscopically for abnormalities. The animal carcasses and tumor tissues were placed in a carcass storage refrigerator for uniform processing.

[0053] The relative tumor weight is calculated as follows: RTW = W T / W C x 100% (wherein WC represents the tumor weight of the solvent control group; WT represents the tumor weight of the treatment group).

[0054] Data are expressed as mean ± SEM (standard error of mean) unless otherwise stated. The T-test was used for the comparison between two groups, and one-way ANOVA was used for the comparison among three or more groups. If the variance was not equal (F value was significantly different), the Games-Howell method was used for the test. If the variance was equal (F value was not significantly different), the Tukey method was used for the analysis. p < 0.05 indicates a significant difference, and p < 0.01 indicates a very significant difference.

[0055] 1.2 Experimental results

[0056] According to the existing literature reports and research, the maximum ineffective dose of 50 mg of apatinib and the maximum ineffective dose of 1 mg of compound 10 were used for administration; paclitaxel was administered at a dose with certain anti-tumor effect as a positive control. The experimental results are shown in Figure 1. The experimental results show the inactive dose of compound 10 and apatinib alone, and the RTW value (relative tumor weight) after the combination with different low-dose chemotherapy drugs. Compound 10 and apatinib alone do not show obvious anti-tumor effect, and the chemotherapy drugs administered at a lower effective dose are also difficult to inhibit the tumor to the desired extent. However, after the combination of the chemotherapy drugs with compound 10, the anti-tumor activity of the chemotherapy drugs is significantly enhanced, and apatinib as a control drug does not show a sensitization effect on the chemotherapy drugs at a lower effective dose.

[0057]

Example 2

[0058] The purpose of this example is to investigate the sensitization effect of compound 10 of formula (I) on the chemotherapy drug oxaliplatin.

[0059] 2.1 Experimental method

[0060] The required number of female BALB / c mice were raised in the SPF animal room. The temperature in the animal room was 20-25°C, the relative humidity was 40%-70%, the light and dark illumination was 12 hours each, and the animals were free to drink water and eat food.

[0061] Dosing and cell lines reference (Sec62 promotes stemness and chemoresistance of human colorectal cancer through activating Wnt / β-catenin pathway) DLD-1 human colorectal adenocarcinoma epithelial cells, from the American Type Culture Collection. Cell culture methods and model preparation methods refer to Example 1. When the tumor volume reaches about 120 mm^3, the animals are randomly grouped according to tumor volume, 10 animals per group. Measure the tumor diameter twice a week after the start of the experiment, at the same time weigh the animal body weight and record, after the completion of the experiment, take the tumor to weigh the tumor weight.

[0062] The day of grouping began to give medicine, after 35 days, the experiment ended. Animal grouping and dosing regimens are as follows, paclitaxel is administered by tail vein, once a week, a total of 5 weeks of administration. Other drugs are administered by gavage, the administration volume is 10 mL / kg, once a day, a total of 5 weeks of administration. The dosing is shown in Table 3.

[0063] Table 3

[0064] Oxaliplatin administration preparation: according to the instructions, dilute to the required concentration according to the administration requirements.

[0065] Other sample administration preparations are prepared according to the method of Example 1.

[0066] Compound 10, apatinib, the inactive dose used alone and different low-dose oxaliplatin used in combination.

[0067] On the 36th day, measure the tumor diameter and weigh the animal body weight, and euthanize the animals. The tumor mass is stripped and photographed, weighed. Grossly dissect the animals and observe the internal organs for abnormalities. Animal carcasses and tumor tissues are placed in the carcass storage refrigerator for uniform processing.

[0068] Relative tumor calculation and data processing methods refer to Example 1.

[0069] 2.2 Experimental results

[0070] According to the existing literature reports and research, the maximum anti-tumor ineffective dose of apatinib is 50 mg, the maximum ineffective dose of compound 10 is 1 mg, and the low effective dose of oxaliplatin is used for administration. The experimental results are shown in Figure 2. The experimental results show that apatinib and compound 10 used alone do not show obvious anti-tumor effect, and the low effective dose of oxaliplatin is also difficult to inhibit the tumor to the desired extent. However, after the combination of oxaliplatin and compound 10, the anti-tumor activity of oxaliplatin is significantly enhanced. Apatinib as a control drug does not show sensitization effect on oxaliplatin at its low effective dose.

[0071] Example 3: Study on the sensitization effect of compound of formula (I) on 5-fluorouracil

[0072] The purpose of this example is to investigate the sensitization effect of compound 10 of formula (I) on the chemotherapeutic drug 5-fluorouracil (5-Fu).

[0073] 3.1 Experimental method

[0074] The model making method is referred to Example 2.

[0075] The drug administration started on the day of grouping and ended after 35 days. The animal grouping and drug administration scheme are shown below. Paclitaxel was administered by tail vein, once a week, for a total of 3 weeks. Other drugs were administered by gavage, and the administration volume was 10 mL / kg, once a day, for a total of 3 weeks. The administration dose is shown in Table 4.

[0076] Table 4

[0077] 5-Fu administration preparation: prepare according to the instructions and dilute to the required concentration according to the administration requirements.

[0078] The administration preparations of other samples are prepared according to the method of Example 1.

[0079] On the 36th day, the tumor diameter was measured, the animal body weight was weighed, and the animal was euthanized. The tumor mass was stripped and photographed, weighed. The animal was subjected to gross dissection, and the internal organs were observed for abnormalities. The animal carcass and tumor tissue were placed in the carcass storage refrigerator for uniform processing.

[0080] The relative tumor calculation and data processing method is referred to Example 1.

[0081] 3.2 Experimental results

[0082] The maximum anti-tumor ineffective dose of apatinib is 50 mg, the maximum ineffective dose of compound 10 is 1 mg, and the 5-Fu is less than the effective dose. The experimental results are shown in Figure 3. The experimental results show that the ineffective dose of compound 10 and apatinib alone does not show obvious anti-tumor effect, and the 5-Fu less than the effective dose is difficult to inhibit the tumor to the desired extent, and after the combination of 5-Fu and compound 10, the anti-tumor activity of 5-Fu is significantly enhanced. Apatinib as a control drug does not show sensitization effect on 5-Fu at its less than effective dose.

[0083] [Example 4] Study on the inhibition of different kinases by the compound of formula (I)

[0084] The purpose of this example is to investigate the inhibition of different kinases by the compounds of formula (I) 1-24.

[0085] 4.1 Experimental method

[0086] The Mobility Shift Assay, Lance Ultra Assay, ADP-Glo Assay and other methods are used to test the inhibition IC50 of different compounds on different kinases under the condition of Km ATP.

[0087] The initial concentration of compound 10 is 10 μM, and the dilution is 10 times, and the compound is tested in duplicate. According to different kinase targets, compounds such as staurosporine, SB431542, GW5074, PI103 are used as standard controls.

[0088] According to the kit instructions, prepare the kinase buffer, termination solution, kinase buffer, enzyme solution and other related solutions.

[0089] The final concentration of the compound is 10 μM, which is prepared to 100 times the concentration, i.e. 1000 μM, and then diluted by a factor of 10. According to the kit operation, test with a fluorescence microplate reader (excitation wavelength 340 nm, test wavelength 520 nm, 495 nm).

[0090] The numerical value ratio of the fluorescence reading (R(520nm / 495nm))

[0091] The above data is converted to the inhibition percentage % = Rmax / (Rmax-Rmin)*100 by the formula

[0092] “min” is the reading of the control well without enzyme; “max” is the reading of the blank solvent added.

[0093] IC 50Results were calculated using the fitted equation: Y = Bottom + (Top-Bottom) / (1+(IC50 / X)^HillSlope).

[0094] 4.2 Experimental results

[0095] The results of the kinase inhibition assay showed that the compounds of the present series have lower inhibitory effect on kinases BRAF, BRAF V600E, AKT1 / 2 / 3, ALK, ALK[G1202R], ALK[L1196M], ALK[C1156Y], ALK[F1174L], ALK[G1269A], ALK[R1275Q], ALK4 / 5, CDK1 / cyclinB, CDK2 / CycA2, CDK2 / CycE1, CDK4 / CycD3, CDK6 / CycD1, CDK6 / CycD3, CDK7 / cyclinH / MAT1, CDK9 / cyclinT1, FGFR1 / 2 / 3 / 4, HER2 / 4, IGF1R, PI3Ka / b / d / g, with IC 50 values all >1000 nM. The compounds of the present series have better inhibitory effect on kinases vEGFR, MET, TRK-A / B / C, with IC 50 values all <100 nM), as shown in Figure 4.

[0096] According to the reported IC50 of apatinib on vEGFR is less than 10 nM, and according to the results of the kinase inhibition assay of the compound of formula (I) above, it also has inhibitory effect on vEGFR. The aforementioned in vivo study of animals showed that apatinib failed to play a sensitization effect relying on the inhibitory effect on the kinase. Therefore, although the compounds of the present series can inhibit vEGFR, their contribution to the sensitization effect is still questionable.

[0097] Since the compounds of the present series have inhibitory effect on MET and TRK kinases, it is necessary to further determine whether the two kinases have strong support for the sensitization of chemotherapy.

[0098]

Example 5

[0099] The purpose of the present example is to investigate the sensitization effect of MET inhibitor compound 10 of formula (I) and carmotinib on the chemotherapeutic drug paclitaxel, respectively.

[0100] 5.1 Experimental method

[0101] The model making method is referred to Example 1.

[0102] Referring to FDA public information [MULTI-DISCIPLINE REVIEW, APPLICATION NUMBER: 213591Orig1s000], a mouse embryo fibroblast NIH3T3 cell line sensitive to carmotinib was used to establish a transplanted tumor model.

[0103] The drug administration was started on the day of grouping and ended after 14 days. The animal grouping and drug administration scheme are shown below. Paclitaxel was administered via tail vein, once a week, for a total of 2 weeks. Other drugs were administered by gavage, with a drug administration volume of 10 mL / kg, once a day, for a total of 2 weeks. The drug administration doses are shown in Table 5.

[0104] Table 5

[0105] Preparation of carmotinib administration preparation: prepared according to the instructions and diluted to the required concentration according to the drug administration requirements.

[0106] The administration preparation of other samples was prepared according to the method of Example 1.

[0107] On the 15th day, the tumor diameter was measured and the animal body weight was weighed, and the animals were euthanized. The tumor mass was stripped and photographed, weighed. The animals were subjected to gross dissection, and the internal organs were observed for abnormalities. The animal carcasses and tumor tissues were placed in the carcass storage refrigerator for uniform processing.

[0108] The relative tumor calculation and data processing method is referred to Example 1.

[0109] 5.2 Experimental results

[0110] The experimental results are shown in Figure 5. Carmotinib is a MET kinase inhibitor, and when used in combination with paclitaxel, it does not show a sensitization effect on the chemotherapeutic drug paclitaxel. However, after the combination of compound 10 and paclitaxel, a good sensitization effect is shown, suggesting that the sensitization effect of the compound is less related to the inhibition of MET kinase.

[0111]

Example 6

[0112] The purpose of this example is to investigate the sensitization effect of TRK inhibitor compound 10 and entrectinib (Entrectinib) on the chemotherapeutic drug paclitaxel, respectively.

[0113] 6.1 Experimental method

[0114] The model making method is referred to Example 1.

[0115] Reference FDA public information [MULTI-DISCIPLINE REVIEW, APPLICATION NUMBER: 212725Orig1s000, 212726Orig1s000], KM12 cell line sensitive to entrectinib was used to establish a transplanted tumor model.

[0116] The drug administration was started on the same day of grouping and ended after 14 days. The animal grouping and drug administration scheme are shown below. Paclitaxel was administered via tail vein, once a week, for a total of 2 weeks. Other drugs were administered by gavage, once a day, with a volume of 10 mL / kg, for a total of 2 weeks. The drug administration doses are shown in Table 6.

[0117] Table 6

[0118] The drug administration preparation of entrectinib and other samples was prepared according to the method of Reference Example 1.

[0119] On the 15th day, the tumor diameter was measured and the animal body weight was measured, and the animals were euthanized. The tumor mass was stripped and photographed and weighed. The animals were subjected to gross autopsy, and the internal organs were observed for abnormalities. The animal carcasses and tumor tissues were placed in the carcass storage refrigerator for uniform processing.

[0120] The relative tumor calculation and data processing method is referred to Reference Example 1.

[0121] 6.2 Experimental results

[0122] The experimental results are shown in Figure 6. Entrectinib is a TRK kinase inhibitor, and when used in combination with paclitaxel, it does not show a sensitization effect on the chemotherapeutic drug paclitaxel. Compound 10 has a good sensitization effect when used in combination with paclitaxel, suggesting that the sensitization effect of the compound is less related to the inhibition of TRK kinase. Moreover, in clinical use, TRK kinase inhibitors are less commonly used in combination with chemotherapeutic drugs, and it is not possible to effectively infer the sensitization potential of the corresponding kinase inhibitors on chemotherapeutic drugs.

[0123]

Example 7

[0124] The purpose of this example is to investigate the sensitization effect of compounds 1-24 represented by formula (I) on the chemotherapeutic drug paclitaxel, respectively.

[0125] 7.1 Experimental method

[0126] Reference Example 1, the sensitization effect of compounds 1-24 in compounds shown in formula (I) on paclitaxel was tested. Since there are many compounds, the test was divided into 4 times, and compound 10 has been verified by other examples, so as to save experimental resources, the sensitization effect of compound 10 was not repeated in this experiment. When each compound is used alone, a lower dose is used to avoid its own antitumor activity as much as possible.

[0127] 7.2 Experimental results

[0128] The experimental results are shown in Figure 7. Each compound has different degrees of sensitization effect on the chemotherapeutic drug paclitaxel.

[0129]

Example 8

[0130] The purpose of this example is to investigate the sensitization effect of compound 10 shown in formula (I) on paclitaxel at different doses.

[0131] 8.1 Experimental method

[0132] The modeling administration was performed according to the method of Reference Example 1.

[0133] The administration started on the day of grouping and ended after 21 days. The animal grouping and administration scheme are shown below. Paclitaxel was administered by tail vein, once a week, for a total of 3 weeks. Other drugs were administered by gavage, with a volume of 10 mL / kg, once a day, for a total of 3 weeks. The administration dose is shown in Table 7.

[0134] Table 7

[0135] Paclitaxel administration preparation: prepare according to the instructions and dilute to the required concentration according to the administration requirements.

[0136] The administration preparation of compound 10 is prepared to the required concentration according to the method disclosed in Chinese patent CN 113336768 B.

[0137] On the 22nd day, the tumor diameter was measured and the animal body weight was weighed, and the animal was euthanized. The tumor mass was stripped and photographed, weighed. The animal was subjected to gross dissection, and the internal organs were observed under a microscope. The animal carcass and tumor tissue were placed in a carcass storage refrigerator for uniform processing.

[0138] The data processing method is referred to Reference Example 1.

[0139] 8.2 Experimental results

[0140] The experimental results are shown in Figure 8. Compound 10 alone has no significant anti-tumor effect, and paclitaxel combined with different doses of compound 10 shows certain chemotherapy sensitization effect, and the intensity of the sensitization effect has a significant dose-dependent, suggesting that the sensitization effect is related to the compound. The results show that in the dose range of 0.01 mg / kg to 1 mg / kg, the compound has a significant sensitization effect on the chemotherapy drugs.

Claims

1. A combined pharmaceutical composition for treating a tumor, comprising, The combination composition comprises a chemotherapeutic drug and at least one compound as shown in formula (I), wherein R1is selected from the following substituents: carboxy-substituted C3-C8alkylacyl, substituted or unsubstituted phosphono, R2 is selected from the group consisting of hydrogen, halogen, R5-substituted C1-C8 alkyl, R5-substituted C6-C12 aryl; R3, R4 are selected from the group consisting of hydrogen, R5-substituted C1-C6 alkyl, or R3, R4 form a five- to twelve-membered aliphatic heterocycle; R5 is selected from the group consisting of hydrogen, 1-3 halogen, hydroxyl, C1-C6 alkoxy, C6-C12 aryl.

2. The combination of claim 1, wherein The compound is selected from:

3. The combination of claim 1, wherein The chemotherapy drug is selected from at least one of the drugs affecting nucleic acid biosynthesis, the drugs affecting DNA structure and function, the drugs interfering with transcription process and preventing RNA synthesis, the drugs inhibiting protein synthesis and function, and the drugs regulating hormone balance in the traditional chemotherapy drugs.

4. The combination of claim 1, wherein The chemotherapy drug is selected from at least one of cyclophosphamide, fluorouracil, adriamycin, daunorubicin, epirubicin, actinomycin, carboplatin, cisplatin, oxaliplatin, vinblastine, camptothecin, paclitaxel, docetaxel, medroxyprogesterone, and tamoxifen.

5. The combination of claim 1, wherein The chemotherapy drug is selected from at least one of oxaliplatin, paclitaxel, and 5-fluorouracil.

6. The combination composition according to claim 1 or claim 5, wherein The compound is compound 10.

7. The combination of claim 1, wherein The dosage form of the combination composition is injection.

8. Use of a compound as shown in formula (I) in the manufacture of a medicament for treating a tumor, wherein, The active ingredient of the drug is a chemotherapy drug, and the compound enhances the efficacy of the chemotherapy drug. ​ 9. Use of the combination composition of any one of claims 1-7 in the preparation of a drug for treating tumors.

10. Use of the compound of formula (I) and the chemotherapeutic agent in the preparation of a medicament for treating tumors, wherein the chemotherapeutic agent is selected from paclitaxel, oxaliplatin, and 5-fluorouracil, and the compound of formula (I) has the following structural formula:

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