Use of a pharmaceutical composition with GL-V9 and a chemotherapeutic drug as active ingredients in treating tumors

Abstract

The application discloses application of a pharmaceutical composition with GL-V9 and a chemotherapeutic drug as active ingredients in treatment of tumors, and the GL-V9 can obviously achieve a drug synergistic effect when the GL-V9 is combined with a classical chemotherapeutic drug to treat tumors, and the effect is obviously superior to combination of similar drugs and the chemotherapeutic drug. The chemotherapeutic drugs include PARP1 inhibitors, capecitabine, irinotecan, sorafenib, cisplatin or 5-fluorouracil; and the tumor types include leukemia, colorectal cancer and lung cancer.

Description

Technical Field

[0001] This invention relates to the application of a pharmaceutical composition, specifically to the application of a pharmaceutical composition with GL-V9 and a chemotherapy drug as active ingredients in the treatment of tumors. Background Technology

[0002] Due to their potential for clinical application in cancer treatment, HSP90 inhibitors are not only constantly being developed into new drugs, but their combined use with chemotherapy drugs to improve the efficacy of cancer treatment is also an important aspect of research and application. Currently, the following combinations of HSP90 inhibitors have been disclosed: onalespib combined with a PARP1 inhibitor for the treatment of breast cancer [Reference: PMID: 34887522], 17-AAG combined with cisplatin for the treatment of ovarian cancer [Reference: PMID: 33331136], etc. However, the research on these HSP90 inhibitors has limited their clinical application due to certain limitations. For example, the research progress on 17-AAG has been slowed due to its poor water solubility and low bioavailability; and the research on Ganetespib has even been considered futile. Therefore, the development of novel HSP90 inhibitors is of great significance for advancing their clinical treatment and application.

[0003] GL-V9 is a novel flavonoid compound that has shown good therapeutic effects on various tumors and has been identified as a novel HSP90 inhibitor [Reference: PMID: 33143000]. However, no studies have yet revealed whether GL-V9, as an HSP90 inhibitor, can have a synergistic effect with chemotherapy drugs. Therefore, this invention, based on the aforementioned existing research techniques on HSP90, compares the application of GL-V9 in combination with chemotherapy drugs in various tumors, revealing its effectiveness and superiority. Summary of the Invention

[0004] Objective of the Invention: The objective of this invention is to provide a compound pharmaceutical composition with GL-V9 and commonly used clinical chemotherapy drugs as active ingredients. Another objective of this invention is to provide the application of the said pharmaceutical composition in leukemia, lung cancer, and colorectal cancer.

[0005] Technical solution: A pharmaceutical composition with HSP90 inhibitors and chemotherapy drugs as active ingredients.

[0006] The pharmaceutical composition wherein the HSP90 inhibitor is GL-V9, Onalespib, Luminespib, 17-AAG, or Ganetespib; and the chemotherapeutic agent is a PARP1 inhibitor, capecitabine, irinotecan, sorafenib, cisplatin, or 5-fluorouracil.

[0007] The pharmaceutical composition wherein the GL-V9 comprises a GL-V9 compound or a pharmaceutically acceptable salt thereof.

[0008] The application of the pharmaceutical composition in the preparation of a drug for treating tumors, wherein the tumors are leukemia, lung cancer, and colorectal cancer.

[0009] The application of GL-V9 in combination with chemotherapy drugs in the preparation of drugs for treating leukemia. The chemotherapy drugs are PARP1 inhibitors, capecitabine, irinotecan, sorafenib, cisplatin, or 5-fluorouracil.

[0010] The application of GL-V9 in combination with chemotherapy drugs in the preparation of drugs for treating lung cancer. The chemotherapy drugs are PARP1 inhibitors, capecitabine, irinotecan, sorafenib, cisplatin, or 5-fluorouracil.

[0011] The application of GL-V9 in combination with chemotherapy drugs in the preparation of drugs for treating colorectal cancer. The chemotherapy drugs are PARP1 inhibitors, capecitabine, irinotecan, sorafenib, cisplatin, or 5-fluorouracil.

[0012] Application of Ganetespib and 5-fluorouracil in the preparation of drugs for treating colon cancer.

[0013] The use of 17-AGG in combination with cisplatin, Ganettespib and 5-fluorouracil, Luminespib and capecitabine, PARP1 inhibitors and Onelespib, Luminespib and irinotecan, and Luminespib and sorafenib in the preparation of drugs for the treatment of leukemia.

[0014] Application of 17-AGG in combination with cisplatin, PARP1 inhibitors and Onelespib in the preparation of drugs for treating lung cancer.

[0015] The pharmaceutical composition according to claim 1 is formulated by adding pharmaceutically acceptable excipients.

[0016] The pharmaceutical composition, wherein the dosage form of the preparation includes tablets, capsules, powders, powder for injection, granules, powders, oral liquids, and syrups.

[0017] Preferably, a pharmaceutical composition comprises GL-V9 and a chemotherapeutic drug as active ingredients. The chemotherapeutic drug is a PARP1 inhibitor, capecitabine, irinotecan, sorafenib, cisplatin, or 5-fluorouracil.

[0018] Application of GL-V9 in the preparation of chemotherapy drug enhancers.

[0019] Beneficial Effects: Compared with existing technologies, this invention has the following advantages: GL-V9, when used in combination with several chemotherapeutic drugs (PARP1 inhibitors, capecitabine, irinotecan, sorafenib, cisplatin, or 5-fluorouracil), can exert a synergistic effect in the treatment of leukemia, colorectal cancer, and lung cancer. At concentrations / doses where GL-V9 or other HSP90 inhibitors are ineffective or have only mild effects on tumors when used alone, the synergistic effect of GL-V9 with the aforementioned chemotherapeutic drugs is significantly stronger than that of other HSP90 inhibitors. Compound drug compositions with GL-V9 and chemotherapeutic drugs as active ingredients show promising application prospects. Detailed Implementation

[0020] 1.1 Experimental Materials

[0021] 1.1.1 Reagents and Materials

[0022] ① compounds

[0023] GL-V9, provided by China Pharmaceutical University, is a pale yellow powder with a purity >95%. Before use, the compound powder should be prepared into a 0.01M stock solution using dimethyl sulfoxide and stored at -80℃. When using, dilute to the required concentration using the appropriate cell culture medium. The structural formula of GL-V9 is as follows:

[0024]

[0025] HSP90 inhibitors: Onalespib, Luminespib, 17-AAG and Ganetspib were purchased from MedChemExpress.

[0026] Chemotherapy drugs: PARP1 inhibitors, capecitabine, irinotecan, sorafenib, cisplatin, and 5-fluorouracil were purchased from MedChemExpress.

[0027] MTT: Yellow powder, purchased from Yisheng Biotechnology.

[0028] ②Cell line

[0029] Human leukemia cell lines Jurkat, ME1 and THP1, colorectal cancer cell line (HCT116) and lung cancer cell line (H1975) were all purchased from the Shanghai Institute of Cell Biology, Chinese Academy of Sciences.

[0030] ③ Cell culture reagents

[0031] RPMI-1640, IMDM, and DMEM liquid culture media were purchased from Jiangsu Kaiji Biotechnology Co., Ltd. Fetal bovine serum (GIBCO, Carlsbad, CA, USA) was stored at -20℃, inactivated in a 56℃ water bath for 35 min before use, aliquoted, and stored at 4℃ for one week. When using, the fetal bovine serum was mixed with the culture medium at a ratio of 1:10.

[0032] 1.1.2 Experimental Apparatus

[0033] YJ-875 Medical Clean Bench (Clean Equipment Factory, Suzhou, China); 3111 Water-jacketed CO2 Incubator (ThermoFisher Scientific, Waltham, MA, USA); 702 Ultra-low Temperature Freezer (ThermoFisher Scientific); Electronic Balance (Sartorius Instrument Systems Co., Ltd., Beijing, China); QIUJING Hemocytometer (Qiujing Biochemical Reagent Instrument Co., Ltd., Shanghai, China); LD4-2 General Centrifuge (Medical Centrifuge Factory, Shanghai, China); 5417R Benchtop Refrigerated High-Speed ​​Centrifuge (Eppendorf AG, Hamburg, Germany); Research Single-Channel Adjustable Pipette (Eppendorf AG); THZ-312 Benchtop Thermostatic Shaker (Jinghong Test Equipment Co., Ltd., Shanghai, China); Varioskan Full-Wavelength Microplate Reader (ThermoFisher Scientific).

[0034] 1.2 Experimental Methods

[0035] 1.2.1 Cell Viability Assay: The principle is that MTT can be reduced by mitochondrial dehydrogenases in cells to form blue-purple formazan crystals. The optical density (OD) value at 570 nm is measured and compared to reflect the relative level of cell viability. The procedure is as follows: 100 μL of cell suspension (5000 cells / well) is uniformly cultured in a 96-well plate. For suspension cells, 50 μL of a specific concentration of HSP90 inhibitor / GL-V9 and 50 μL of a specific concentration of chemotherapeutic agent are added immediately after cell addition; for adherent cells, the two drugs are added after cell adhesion and growth, finally reaching the final therapeutic concentration. Cells are cultured in a 37℃, 5% CO2 incubator; after 48 h of drug treatment, 15 μL of MTT solution is added to each well. After incubation in the incubator for 3 h, the OD value at 570 nM is measured using a microplate reader to calculate cell viability before and after drug administration.

[0036] 1.2.2 Combination Index (CI) Analysis: The intermediate-effect principle was adopted, and the CI was calculated using Compusyn software. The average CI interval and interaction evaluation are shown in Table 1.

[0037] Table 1. Average CI value range and interaction evaluation of compound synergistic effects

[0038] CI Joint effect logo ＜0.1 The synergistic effect is very strong. +++++ 0.1 ≤ CI < 0.3 Strong synergistic effect ++++ 0.3 ≤ CI < 0.7 Strong synergistic effect +++ 0.7 ≤ CI < 0.85 Synergistic effect is moderate ++ 0.85 ≤ CI < 0.90 Slight synergistic effect + 0.90 ≤ CI < 1.10 Additive effect ± 1.10 ≤ CI < 1.20 Mild antagonistic effect - 1.20 ≤ CI < 1.45 Moderate antagonistic effect -- 1.45 ≤ CI < 3.3 Strong antagonistic effect --- 3.3≤CI<10 Strong antagonistic effect ---- CI > 10 The antagonistic effect is very strong. -----

[0039] Example 1

[0040] Synergistic effect of GL-V9 combined with cisplatin in the treatment of leukemia

[0041] Using three representative leukemia cell types—Jurkat, THP1, and ME1—as research subjects, this study compared the therapeutic effects of the publicly available HSP90 inhibitor 17-AGG and cisplatin in suppressing leukemia. Table 2 shows the inhibitory levels of GL-V9, 17-AGG, and cisplatin on leukemia cells under their individual effects (0 < Effect < 1). Table 3 shows the inhibition rate and combined effect index of 17-AGG and cisplatin on leukemia cells after combined administration; Table 4 shows the inhibition rate and combined effect index of GL-V9 and cisplatin on leukemia cells after combined administration.

[0042] Table 2. Inhibitory effects of GL-V9, 17-AGG, and cisplatin alone on Jurkat cells.

[0043]

[0044] Table 3: Combination effect index (CI) analysis of 17-AAG combined with cisplatin on leukemia cells Jurkat

[0045] Cisplatin (μM) 17AAG (μM) Effect CI Joint effect 1.0 0.01563 0.2825 0.26933 Strong synergistic effect 1.0 0.03125 0.2195 0.52767 Strong synergistic effect 1.0 0.0625 0.3114 0.38380 Strong synergistic effect 1.0 0.125 0.3206 0.56376 Strong synergistic effect 1.0 0.25 0.2863 1.20981 Moderate antagonistic effect 2.0 0.01563 0.26979 0.51620 Strong synergistic effect 2.0 0.03125 0.2872 0.52214 Strong synergistic effect 2.0 0.0625 0.35115 0.43389 Strong synergistic effect 2.0 0.125 0.35872 0.57412 Strong synergistic effect 2.0 0.25 0.30943 1.21065 Moderate antagonistic effect 4.0 0.01563 0.31293 0.72610 Synergistic effect is moderate 4.0 0.03125 0.31760 0.75668 Synergistic effect is moderate 4.0 0.0625 0.38463 0.57709 Strong synergistic effect 4.0 0.125 0.33605 0.95037 Additive effect 4.0 0.25 0.31670 1.48449 Strong antagonistic effect

[0046] As shown in Table 3, the combined use of 17-AAG and cisplatin for the treatment of leukemia showed the following synergistic effects: cisplatin 1.0 μM, 17-AAG at 0.01563–0.125 μM exhibited a strong or relatively strong synergistic effect; cisplatin 2.0 μM, 17-AAG at 0.01563–0.125 μM exhibited a relatively strong synergistic effect; and cisplatin 4.0 μM, 17-AAG at 0.01563–0.0625 μM exhibited a moderate or relatively strong synergistic effect.

[0047] Table 4: Combination effect index (CI) analysis of GL-V9 combined with cisplatin on leukemia cells Jurkat

[0048] Cisplatin (μM) GL-V9 (μM) Effect CI Joint effect 1.0 0.25 0.27297 0.27406 Strong synergistic effect 1.0 0.5 0.26445 0.34787 Strong synergistic effect 1.0 1.0 0.51466 0.13809 Strong synergistic effect 1.0 2.0 0.30701 0.55816 Strong synergistic effect 1.0 4.0 0.56239 0.32139 Strong synergistic effect 2.0 0.25 0.3035 0.40581 Strong synergistic effect 2.0 0.5 0.357 0.33557 Strong synergistic effect 2.0 1.0 0.39930 0.33337 Strong synergistic effect 2.0 2.0 0.42507 0.41192 Strong synergistic effect 2.0 4.0 0.65677 0.24083 Strong synergistic effect 4.0 0.25 0.51773 0.23421 Strong synergistic effect 4.0 0.5 0.46856 0.32784 Strong synergistic effect 4.0 1.0 0.56770 0.23145 Strong synergistic effect 4.0 2.0 0.51693 0.38063 Strong synergistic effect 4.0 4.0 0.69336 0.24227 Strong synergistic effect

[0049] As shown in Table 4, GL-V9 and cisplatin have a synergistic effect in the treatment of leukemia, with cisplatin at 1.0-4.0 μM and GL-V9 at 0.25-4.0 μM.

[0050] In summary, these results confirm that GL-V9 combined with cisplatin has a strong or relatively strong synergistic effect on Jurkat leukemia cells, and the overall combined effect is stronger than that of 17-AAG combined with cisplatin, demonstrating the advantages of GL-V9 combined with cisplatin in the preparation of drug compositions for the treatment of leukemia.

[0051] Example 2

[0052] Synergistic effect of GL-V9 combined with 5-fluorouracil in the treatment of leukemia

[0053] Using three representative leukemia cell types—Jurkat, THP1, and ME1—as research subjects, this study compared the combined use of the publicly disclosed HSP90 inhibitor Ganettespib and 5-fluorouracil to investigate the therapeutic effect of the combined use of GL-V9 and 5-fluorouracil on leukemia cell inhibition. Table 5 shows the inhibitory levels of GL-V9, Ganettespib, and 5-fluorouracil on leukemia cells under their individual effects (0 < Effect < 1). Table 6 shows the inhibition rate and combined effect index of Ganettespib and 5-fluorouracil on leukemia cells after combined administration; Table 7 shows the inhibition rate and combined effect index of GL-V9 and 5-fluorouracil on leukemia cells after combined administration.

[0054] Table 5. Inhibitory effects of GL-V9, Ganetspib, and 5-fluorouracil on Jurkat cells under individual action.

[0055]

[0056] Table 6: Combination effect index (CI) analysis of Ganetspib combined with 5-fluorouracil on Jurkat leukemia cells

[0057] 5-Fluorouracil (μM) Ganetespib (μM) Effect CI Joint effect 2.0 0.01563 0.17027 2.15653 Strong antagonistic effect 2.0 0.03125 0.42929 0.61966 Strong synergistic effect 2.0 0.0625 0.51537 0.65611 Strong synergistic effect 2.0 0.125 0.57726 0.83076 Synergistic effect is moderate 2.0 0.25 0.62855 1.14574 Mild antagonistic effect 4.0 0.01563 0.26660 1.37258 Moderate antagonistic effect 4.0 0.03125 0.49022 0.58437 Strong synergistic effect 4.0 0.0625 0.56480 0.60070 Strong synergistic effect 4.0 0.125 0.64043 0.64470 Strong synergistic effect 4.0 0.25 0.60477 1.41301 Moderate antagonistic effect 8.0 0.01563 0.41865 0.97090 Additive effect 8.0 0.03125 0.50197 0.81466 Synergistic effect is moderate 8.0 0.0625 0.56843 0.79164 Synergistic effect is moderate 8.0 0.125 0.65394 0.73652 Synergistic effect is moderate 8.0 0.25 0.62014 1.45261 Strong antagonistic effect

[0058] As shown in Table 6, the combined use of Ganettespib and 5-fluorouracil for the treatment of leukemia showed a strong to moderate synergistic effect when 5-fluorouracil was at 2.0 μM and Ganettespib was at 0.03125–0.125 μM. At 4.0 μM, Ganettespib showed a strong synergistic effect when 5-fluorouracil was at 0.03125–0.125 μM. At 8.0 μM, Ganettespib showed a moderate synergistic effect when 5-fluorouracil was at 0.03125–0.125 μM.

[0059] Table 7: Combination effect index (CI) analysis of GL-V9 combined with 5-fluorouracil on leukemia cells Jurkat

[0060] GL-V9 (μM) 5-Fluorouracil (μM) Effect CI Joint effect 0.25 2.0 0.25073 0.45310 Strong synergistic effect 0.25 4.0 0.49266 0.29620 Strong synergistic effect 0.25 8.0 0.63750 0.31756 Strong synergistic effect 0.5 2.0 0.23677 0.55523 Strong synergistic effect 0.5 4.0 0.44384 0.38662 Strong synergistic effect 0.5 8.0 0.64997 0.31334 Strong synergistic effect 1.0 2.0 0.24601 0.65668 Strong synergistic effect 1.0 4.0 0.42957 0.46721 Strong synergistic effect 1.0 8.0 0.58762 0.43810 Strong synergistic effect 2.0 2.0 0.28770 0.74314 Synergistic effect is moderate 2.0 4.0 0.45009 0.53789 Strong synergistic effect 2.0 8.0 0.62127 0.43706 Strong synergistic effect 4.0 2.0 0.62127 0.30327 Strong synergistic effect 4.0 4.0 0.62955 0.37193 Strong synergistic effect 4.0 8.0 0.72879 0.33944 Strong synergistic effect

[0061] As shown in Table 7, the combined use of GL-V9 and 5-fluorouracil for the treatment of leukemia showed synergistic effects at GL-V9 concentrations of 0.25-4.0 μM and 5-fluorouracil concentrations of 2.0-8.0 μM.

[0062] In summary, these results confirm that GL-V9 combined with 5-fluorouracil exhibits a good synergistic effect in the treatment of leukemia, with synergistic effects ranging from strong to moderate, while Ganettespib combined with 5-fluorouracil does not achieve the same effect. These results demonstrate the advantages of GL-V9 combined with 5-fluorouracil in the preparation of drug compositions for the treatment of leukemia.

[0063] Example 3

[0064] Synergistic effect of GL-V9 combined with capecitabine in the treatment of leukemia

[0065] Using three representative leukemia cell types—Jurkat, THP1, and ME1—as research subjects, this study compared the combined use of the publicly available HSP90 inhibitors Luminespib and capecitabine to investigate the therapeutic effect of the combination of GL-V9 and capecitabine in inhibiting leukemia. Table 8 shows the inhibitory levels of GL-V9, Luminespib, and capecitabine on leukemia cells under their individual effects (0 < Effect < 1). Table 9 shows the inhibition rate and combined effect index of Luminespib and capecitabine on leukemia cells after combined administration; Table 10 shows the inhibition rate and combined effect index of GL-V9 and capecitabine on leukemia cells after combined administration.

[0066] Table 8. Inhibitory levels of GL-V9, Luminespib, and capecitabine on Jurkat cells by individual treatment.

[0067]

[0068] Table 9: Combination effect index (CI) analysis of Luminespib combined with capecitabine on Jurkat leukemia cells

[0069]

[0070]

[0071] As shown in Table 9, the combined use of Luminespib and capecitabine for the treatment of leukemia showed a synergistic effect between capecitabine 32.0 μM and Luminespib in the range of 0.01563–0.125 μM. At capecitabine 64.0 μM, the synergistic effect between Luminespib in the range of 0.01563–0.03125 μM was strong or relatively strong. At capecitabine 128.0 μM, the synergistic effect between Luminespib in the range of 0.01563–0.0625 μM was also strong or relatively strong.

[0072] Table 10: Combination effect index (CI) analysis of GL-V9 combined with capecitabine on leukemia cells Jurkat

[0073] GL-V9 (μM) Capecitabine (μM) Effect CI Joint effect 0.25 32.0 0.04652 39.6846 The antagonistic effect is very strong. 0.25 64.0 0.32126 0.04503 The synergistic effect is very strong. 0.25 128.0 0.38032 0.03520 The synergistic effect is very strong. 0.5 32.0 0.06232 1.21844 Moderate antagonistic effect 0.5 64.0 0.21959 0.14745 Strong synergistic effect 0.5 128.0 0.43855 0.05601 The synergistic effect is very strong. 1.0 32.0 0.04865 22.4935 The antagonistic effect is very strong. 1.0 64.0 0.09427 0.76037 Synergistic effect is moderate 1.0 128.0 0.54144 0.07571 The synergistic effect is very strong. 2.0 32.0 0.08768 1.63797 Strong antagonistic effect 2.0 64.0 0.10884 1.30169 Moderate antagonistic effect 2.0 128.0 0.44852 0.21561 Strong synergistic effect 4.0 32.0 0.28352 0.85374 Slight synergistic effect 4.0 64.0 0.35901 0.61417 Strong synergistic effect 4.0 128.0 0.51655 0.33292 Strong synergistic effect

[0074] As shown in Table 10, the combined use of GL-V9 and capecitabine for the treatment of leukemia showed the following synergistic effects: GL-V9 0.25 μM, capecitabine 64.0–128.0 μM, very strong synergistic effect; GL-V9 0.5 μM, capecitabine 64.0–128.0 μM, very strong or moderate synergistic effect; GL-V9 1.0 μM, capecitabine 64.0–128.0 μM, very strong or moderate synergistic effect; GL-V9 2.0 μM, capecitabine 128.0 μM, strong synergistic effect; GL-V9 4.0 μM, capecitabine 32.0–128.0 μM, synergistic effect.

[0075] In summary, these results confirm that GL-V9 combined with capecitabine can produce a very strong synergistic effect within a certain range of proportions. This synergistic strength is significantly better than that of Luminespib combined with capecitabine in the treatment of leukemia, demonstrating the advantages of GL-V9 combined with capecitabine in the preparation of drug compositions for the treatment of leukemia.

[0076] Example 4

[0077] Synergistic effect of GL-V9 combined with PARP1 inhibitors in the treatment of leukemia

[0078] Taking three highly representative leukemia cells, Jurkat, THP1, and ME1, as the research objects, comparing the combined use of the publicly available HSP90 inhibitor Onelespib and the PARP1 inhibitor Olaparib, and studying the therapeutic effect of the combination of GL-V9 and Olaparib in inhibiting leukemia. As shown in Table 11, the inhibitory levels (0 < Effect < 1) of GL-V9, Onelespib, and Olaparib on leukemia cells alone are presented. As shown in Table 12, the inhibition rates and combination indices of the combination of Onelespib and Olaparib on leukemia cells are shown; as shown in Table 13, the inhibition rates and combination indices of the combination of GL-V9 and Olaparib on leukemia cells are shown.

[0079] Table 11 Inhibitory levels of GL-V9, Onelespib, and Olaparib on Jurkat cells alone

[0080]

[0081] Table 12: Analysis of the combination index (CI) of Onelespib combined with Olaparib on leukemia cell Jurkat

[0082]

[0083]

[0084] As shown in Table 12, when Onelespib and Olaparib are used in combination to treat leukemia, Olaparib at 4.0 - 16.0 μM and Onelespib at 0.01563 - 0.25 μM both have a synergistic effect.

[0085] Table 13: Analysis of the combination index (CI) of GL-V9 combined with Olaparib on leukemia cell Jurkat

[0086] GL-V9 (μM) Olaparib (μM) Effect CI Joint effect 0.25 4.0 0.32124 0.25410 Strong synergistic effect 0.25 8.0 0.39718 0.35019 Strong synergistic effect 0.25 16.0 0.61582 0.31641 Strong synergistic effect 0.5 4.0 0.40240 0.22026 Strong synergistic effect 0.5 8.0 0.37793 0.41057 Strong synergistic effect 0.5 16.0 0.57662 0.37931 Strong synergistic effect 1.0 4.0 0.38529 0.30340 Strong synergistic effect 1.0 8.0 0.40506 0.43631 Strong synergistic effect 1.0 16.0 0.54935 0.45325 Strong synergistic effect 2.0 4.0 0.35857 0.48959 Strong synergistic effect 2.0 8.0 0.39104 0.59391 Strong synergistic effect 2.0 16.0 0.51076 0.60239 Strong synergistic effect 4.0 4.0 0.56412 0.36791 Strong synergistic effect 4.0 8.0 0.53314 0.51311 Strong synergistic effect 4.0 16.0 0.73746 0.32244 Strong synergistic effect

[0087] As shown in Table 13, when GL-V9 and Olaparib are used in combination to treat leukemia, GL-V9 at 0.25 - 4.0 μM and Olaparib at 4.0 - 16.0 μM both have a synergistic effect.

[0088] In summary, the results confirm that the combination of GL-V9 or Onelespib with Olaparib can achieve good synergistic effects. It shows the feasibility of the application of the combination of these two drugs in the preparation of a drug composition for treating leukemia.

[0089] Example 5

[0090] Synergistic effect of GL-V9 combined with irinotecan in the treatment of leukemia

[0091] Using three highly representative leukemia cell types—Jurkat, THP1, and ME1—as research subjects, this study compared the combined use of the publicly disclosed HSP90 inhibitors Luminespib and irinotecan to investigate the therapeutic effect of the combined use of GL-V9 and irinotecan in inhibiting leukemia. Table 14 shows the inhibitory levels of GL-V9, Luminespib, and irinotecan on leukemia cells under their individual effects (0 < Effect < 1). Table 15 shows the inhibition rate and combined effect index of Luminespib and irinotecan on leukemia cells after combined administration; Table 16 shows the inhibition rate and combined effect index of GL-V9 and irinotecan on leukemia cells after combined administration.

[0092] Table 14. Inhibitory effects of GL-V9, Luminespib, and irinotecan alone on THP1 cells.

[0093]

[0094] Table 15: Index (CI) analysis of the combined effect of Luminespib and irinotecan on THP1 in leukemia cells.

[0095]

[0096]

[0097] As shown in Table 15, the combined use of Luminespib and irinotecan for the treatment of leukemia showed the following synergistic effects: Luminespib 0.01563 μM, irinotecan 4.0-8.0 μM: strong or strong synergistic effect; Luminespib 0.03125 μM, irinotecan 4.0-8.0 μM: strong or strong synergistic effect; Luminespib 0.0625 μM, irinotecan 4.0-8.0 μM: strong or strong synergistic effect; Luminespib 0.125 μM, irinotecan 4.0-8.0 μM: strong or moderate synergistic effect.

[0098] Table 16: Combination effect index (CI) analysis of GL-V9 combined with irinotecan on THP1 in leukemia cells

[0099] Irinotecan (μM) GL-V9 (μM) Effect CI Joint effect 4.0 0.25 0.29049 0.46153 Strong synergistic effect 4.0 0.5 0.35384 0.45493 Strong synergistic effect 4.0 1.0 0.37243 0.53542 Strong synergistic effect 4.0 2.0 0.34362 0.78626 Synergistic effect is moderate 4.0 4.0 0.33732 1.23575 Moderate antagonistic effect 8.0 0.25 0.50707 0.56889 Strong synergistic effect 8.0 0.5 0.49256 0.61861 Strong synergistic effect 8.0 1.0 0.47264 0.71534 Synergistic effect is moderate 8.0 2.0 0.48091 0.84771 Synergistic effect is moderate 8.0 4.0 0.47880 1.14072 Mild antagonistic effect 16.0 0.25 0.49688 1.12429 Mild antagonistic effect 16.0 0.5 0.49514 1.16226 Mild antagonistic effect 16.0 1.0 0.44627 1.34799 Moderate antagonistic effect 16.0 2.0 0.37515 1.73937 Strong antagonistic effect 16.0 4.0 0.25260 2.89107 Strong antagonistic effect

[0100] As shown in Table 16, GL-V9 and irinotecan were used in combination to treat leukemia. When the concentration of irinotecan was 4.0 μM, the synergistic effect of GL-V9 at 0.25 - 2.0 μM was strong or moderate. When the concentration of irinotecan was 8.0 μM, the synergistic effect of GL-V9 at 0.25 - 2.0 μM was strong or moderate.

[0101] In summary, compared with the strong antagonistic effect produced by the combination of Luminespib and irinotecan at a relatively large concentration ratio, GL-V9 combined with irinotecan does not produce a strong or very strong antagonistic effect within the concentration range of single-drug screening. According to the experimental results, a synergistic effect of inhibiting leukemia can be produced within the range of the GL-V9:irinotecan molar ratio of 1:0.0625 to 1:0.25.

[0102] Example 6

[0103] Synergistic effect of GL-V9 combined with sorafenib in the treatment of leukemia

[0104] Taking three highly representative leukemia cells, Jurkat, THP1, and ME1, as the research objects, comparing the combination of the publicly available technology HSP90 inhibitor Luminespib and sorafenib, and studying the therapeutic effect of the combination of GL-V9 and sorafenib in inhibiting leukemia. As shown in Table 17, the inhibition levels (0 < Effect < 1) of GL-V9, Luminespib, and sorafenib on leukemia cells when acting alone are presented. As shown in Table 18, the inhibition rate and combination index of the combination of Luminespib and sorafenib on leukemia cells are shown; as shown in Table 19, the inhibition rate and combination index of the combination of GL-V9 and sorafenib on leukemia cells are shown.

[0105] Table 17 Inhibition levels of GL-V9, Luminespib, and sorafenib on ME1 cells when acting alone

[0106]

[0107] Table 18: Analysis of the combination index (CI) of Luminespib combined with sorafenib on leukemia cell ME1

[0108]

[0109]

[0110] As shown in Table 18, when Luminespib and sorafenib were used in combination to treat leukemia, with the concentration of Luminespib being 0.01563 μM, the synergistic effect of sorafenib at 4.0 μM was moderate. Table 19: Analysis of the combination index (CI) of GL-V9 combined with sorafenib on leukemia cell ME1

[0111]

[0112]

[0113] As shown in Table 19, the combination of GL-V9 and sorafenib for the treatment of leukemia, with sorafenib at 4.0-16.0 μM and GL-V9 at 0.25-4.0 μM, all showed synergistic effects.

[0114] In summary, these results confirm that, compared to the antagonistic effect of Luminespib combined with sorafenib in the treatment of leukemia, GL-V9 combined with sorafenib can exert a good synergistic effect, demonstrating the advantages of GL-V9 combined with sorafenib in the preparation of drug compositions for the treatment of leukemia.

[0115] Example 7

[0116] Synergistic effect of GL-V9 combined with cisplatin in the treatment of lung cancer

[0117] Using H1975 cells, a highly representative lung cancer cell line, as the research object, this study compared the combined use of the publicly disclosed HSP90 inhibitor 17-AGG and cisplatin to investigate the therapeutic effect of GL-V9 and cisplatin in inhibiting lung cancer. Table 20 shows the inhibitory levels of GL-V9, 17-AGG, and cisplatin on lung cancer cells under their individual effects (0 < Effect < 1). Table 21 shows the inhibition rate and combined effect index of 17-AGG and cisplatin on lung cancer cells after combined administration; Table 22 shows the inhibition rate and combined effect index of GL-V9 and cisplatin on lung cancer cells after combined administration.

[0118] Table 20 Inhibitory levels of H1975 cells by GL-V9, 17-AGG and cisplatin alone

[0119]

[0120] Table 21: Combination effect index (CI) analysis of 17-AAG combined with cisplatin on H1975

[0121]

[0122]

[0123] As shown in Table 21, the combination of 17-AAG and cisplatin for the treatment of lung cancer showed a strong or moderate synergistic effect when 17-AAG was 0.0625 μM and cisplatin was 1.0-2.0 μM.

[0124] Table 22: Combination effect index (CI) analysis of GL-V9 combined with cisplatin on H1975

[0125] Cisplatin (μM) GL-V9 (μM) Effect CI Joint effect 1.0 0.5 0.05482 0.51900 Strong synergistic effect 1.0 1.0 0.23725 0.23582 Strong synergistic effect 1.0 2.0 0.21028 0.31331 Strong synergistic effect 1.0 4.0 0.22893 0.39855 Strong synergistic effect 2.0 0.5 0.07407 0.80679 Synergistic effect is moderate 2.0 1.0 0.20696 0.46250 Strong synergistic effect 2.0 2.0 0.19501 0.54151 Strong synergistic effect 2.0 4.0 0.14605 0.81072 Synergistic effect is moderate 4.0 0.5 0.09556 1.33999 Moderate antagonistic effect 4.0 1.0 0.20096 0.88479 Slight synergistic effect 4.0 2.0 0.13799 1.20156 Moderate antagonistic effect 4.0 4.0 0.16782 1.19882 Mild antagonistic effect

[0126] As shown in Table 22, when GL-V9 and cisplatin are used in combination to treat lung cancer, with cisplatin at 1.0 - 2.0 μM and GL-V9 at 0.5 - 4.0 μM, the synergistic effect is strong, relatively strong or moderate. When cisplatin is at 4.0 μM and GL-V9 is at 1.0 μM, the synergistic effect is slight.

[0127] In summary, the results confirm that within the screened concentration range, GL-V9 combined with cisplatin can produce a relatively obvious synergistic effect on lung cancer cells, while 17-AAG combined with cisplatin mainly produces an additive or antagonistic effect on lung cancer cells. This result indicates the application value of preparing a lung cancer drug composition with GL-V9 combined with cisplatin.

[0128] Example 8

[0129] Synergistic effect of GL-V9 combined with 5-fluorouracil in the treatment of colorectal cancer

[0130] Taking the colorectal cancer representative cell HCT116 cells as the research object, comparing the combination of the publicly available HSP90 inhibitor Ganetespib and 5-fluorouracil, and studying the therapeutic effect of the combination of GL-V9 and 5-fluorouracil in inhibiting colorectal cancer. As shown in Table 23, it is the inhibition level (0 < Effect < 1) of GL-V9, Ganetespib and 5-fluorouracil on colorectal cancer cells when acting alone. As shown in Table 24, it is the inhibition rate and combination index of the combination of Ganetespib and 5-fluorouracil on colorectal cancer cells; as shown in Table 25, it is the inhibition rate and combination index of the combination of GL-V9 and 5-fluorouracil on colorectal cancer cells.

[0131] Table 23 Inhibition level of GL-V9, Ganetespib and 5-fluorouracil on HCT116 cells when acting alone

[0132]

[0133] Table 24: Analysis of the combination index (CI) of Ganetespib combined with 5-fluorouracil on HCT116

[0134] <00​​​As shown in Table 24, the combination of Ganetespib and 5-fluorouracil for the treatment of colorectal cancer, with 5-fluorouracil at 2.0 μM and Ganetespib at 0.01563 - 0.125 μM, has a strong or relatively strong synergistic effect. When 5-fluorouracil is at 4.0 μM and Ganetespib is at 0.01563 - 0.125 μM, the synergistic effect is strong or relatively strong. When 5-fluorouracil is at 8.0 μM and Ganetespib is at 0.01563, 0.0625 - 0.125 μM, the synergistic effect is relatively strong or moderate.

[0137] Table 25: Analysis of the combination index (CI) of GL-V9 combined with 5-fluorouracil on HCT116

[0138] 5-Fluorouracil (μM) GL-V9 (μM) Effect CI Joint effect 2.0 0.5 0.19299 0.16039 Strong synergistic effect 2.0 1.0 0.11146 0.44092 Strong synergistic effect 2.0 2.0 0.99746 0.00487 The synergistic effect is very strong. 2.0 4.0 0.14014 1.21395 Moderate antagonistic effect 2.0 8.0 0.79477 0.32937 Strong synergistic effect 4.0 0.5 0.22382 0.17623 Strong synergistic effect 4.0 1.0 0.19220 0.32208 Strong synergistic effect 4.0 2.0 0.17426 0.59748 Strong synergistic effect 4.0 4.0 0.29574 0.68479 Strong synergistic effect 4.0 8.0 0.81524 0.30526 Strong synergistic effect 8.0 0.5 0.33435 0.14736 Strong synergistic effect 8.0 1.0 0.27998 0.27158 Strong synergistic effect 8.0 2.0 0.26960 0.45640 Strong synergistic effect 8.0 4.0 0.35817 0.60222 Strong synergistic effect 8.0 8.0 0.84194 0.27382 Strong synergistic effect

[0139] As shown in Table 25, the combination of GL-V9 and 5-fluorouracil for the treatment of colorectal cancer, with 5-fluorouracil at 2.0 μM and GL-V9 at 0.5 - 2.0, 8.0 μM, has a strong or very strong synergistic effect. When 5-fluorouracil is at 4.0 - 8.0 μM and GL-V9 is at 0.5 - 8.0 μM, the synergistic effect is strong.

[0140] In summary, compared with the combination of Ganetespib and 5-fluorouracil, the combination of GL-V9 and 5-fluorouracil mainly produces a synergistic effect, and the synergistic effect is very strong, strong and relatively strong, showing advantages in the preparation of drug compositions for colorectal cancer.

[0141] Example 9

[0142] Synergistic effect of GL-V9 combined with capecitabine in the treatment of colorectal cancer

[0143] Taking the representative colorectal cancer cell HCT116 cells as the research object, comparing the combination of the publicly available HSP90 inhibitor Luminespib and capecitabine, and studying the therapeutic effect of the combination of GL-V9 and capecitabine in inhibiting colorectal cancer. As shown in Table 26, it is the inhibition level (0 < Effect < 1) of GL-V9, Luminespib and capecitabine on colorectal cancer cells when acting alone. As shown in Table 27, it is the inhibition rate and combination index of the combination of Luminespib and capecitabine on colorectal cancer cells; as shown in Table 28, it is the inhibition rate and combination index of the combination of GL-V9 and capecitabine on colorectal cancer cells.

[0144] Table 26 Inhibition level of GL-V9, Luminespib and capecitabine on HCT116 cells when acting alone

[0145]

[0146] Table 27: Combination effect index (CI) analysis of Luminespib combined with capecitabine on HCT116

[0147]

[0148]

[0149] As shown in Table 27, the combined use of Luminespib and capecitabine for the treatment of colorectal cancer showed strong or relatively strong synergistic effects at capecitabine 1.0 μM and Luminespib 0.01563-0.125 μM. At capecitabine 2.0 μM and Luminespib 0.01563-0.125 μM, the synergistic effect was relatively strong. At capecitabine 4.0 μM and Luminespib 0.01563-0.0625 μM, the synergistic effect was relatively strong or moderate.

[0150] Table 28: Combination effect index (CI) analysis of GL-V9 combined with capecitabine on HCT116

[0151] Capecitabine (μM) GL-V9 (μM) Effect CI Joint effect 1.0 0.25 0.27297 0.27406 Strong synergistic effect 1.0 0.5 0.26445 0.34787 Strong synergistic effect 1.0 1.0 0.51466 0.13809 Strong synergistic effect 1.0 2.0 0.30701 0.55816 Strong synergistic effect 1.0 4.0 0.56239 0.32139 Strong synergistic effect 2.0 0.25 0.3035 0.40581 Strong synergistic effect 2.0 0.5 0.357 0.33557 Strong synergistic effect 2.0 1.0 0.39930 0.33337 Strong synergistic effect 2.0 2.0 0.42507 0.41192 Strong synergistic effect 2.0 4.0 0.65677 0.24083 Strong synergistic effect 4.0 0.25 0.51773 0.23421 Strong synergistic effect 4.0 0.5 0.46856 0.32784 Strong synergistic effect 4.0 1.0 0.56770 0.23145 Strong synergistic effect 4.0 2.0 0.51693 0.38063 Strong synergistic effect 4.0 4.0 0.69336 0.24227 Strong synergistic effect

[0152] As shown in Table 28, the combined use of GL-V9 and capecitabine for the treatment of colorectal cancer, with capecitabine at 1-4.0 μM and GL-V9 at 0.25-4.0 μM, both showed synergistic effects.

[0153] In summary, these results confirm that, compared with the combination drug prepared by Luminespib and capecitabine for the treatment of colorectal cancer, GL-V9 combined with capecitabine produces a strong and relatively strong synergistic effect on colorectal cancer cells, demonstrating the advantages of GL-V9 combined with capecitabine in the preparation of drug compositions for the treatment of colorectal cancer.

[0154] Example 10

[0155] Synergistic effect of GL-V9 combined with PARP1 inhibitors in the treatment of lung cancer

[0156] Using H1975, a highly representative lung cancer cell line, as the research object, this study compared the combined use of the publicly disclosed HSP90 inhibitor onelespib and the PARP1 inhibitor olaparib to investigate the therapeutic effect of the combination of GL-V9 and olaparib in inhibiting lung cancer. Table 29 shows the inhibitory levels of GL-V9, onelespib, and olaparib on lung cancer cells under their individual effects (0 < Effect < 1). Table 30 shows the inhibition rate and combined effect index of onelespib and olaparib on lung cancer cells after combined administration; Table 31 shows the inhibition rate and combined effect index of GL-V9 and olaparib on lung cancer cells after combined administration.

[0157] Table 29 Inhibitory levels of GL-V9, onelespib, and olaparib on H1975 cells under individual action.

[0158]

[0159] Table 30: Combination effect index (CI) analysis of onelespib combined with olaparib on H1975

[0160]

[0161]

[0162] As shown in Table 30, the combination of onelespib and olaparib for the treatment of lung cancer showed a strong synergistic effect: olaparib 4.0 μM and onelespib 0.125 μM; olaparib 8.0 μM and onelespib 0.01563-0.125 μM; and olaparib 16.0 μM and onelespib 0.03125-0.125 μM.

[0163] Table 31: Combination effect index (CI) analysis of GL-V9 combined with olaparib on H1975

[0164] GL-V9 (μM) Olaparib (μM) Effect CI Joint effect 0.5 4.0 0.17039 0.22114 Strong synergistic effect 0.5 8.0 0.23594 0.30766 Strong synergistic effect 0.5 16.0 0.25297 0.55332 Strong synergistic effect 1.0 4.0 0.15490 0.27601 Strong synergistic effect 1.0 8.0 0.22426 0.34869 Strong synergistic effect 1.0 16.0 0.27021 0.54240 Strong synergistic effect 2.0 4.0 0.16785 0.32673 Strong synergistic effect 2.0 8.0 0.29232 0.31340 Strong synergistic effect 2.0 16.0 0.30514 0.52139 Strong synergistic effect 4.0 4.0 0.19391 0.40997 Strong synergistic effect 4.0 8.0 0.29842 0.38812 Strong synergistic effect 4.0 16.0 0.30096 0.60898 Strong synergistic effect

[0165] As shown in Table 31, the combined use of GL-V9 and Olaparib for the treatment of lung cancer, with GL-V9 at 0.5-4.0 μM and Olaparib at 4.0-16.0 μM, both showed synergistic effects.

[0166] In summary, these results confirm that, compared to the combination drug prepared by onelespib and olaparib for the treatment of lung cancer, GL-V9 combined with olaparib produces a strong and relatively strong synergistic effect on lung cancer cells, demonstrating the advantages of GL-V9 combined with the PARP1 inhibitor olaparib in the preparation of drug compositions for the treatment of lung cancer.

Claims

1. A pharmaceutical composition, characterized in that, The active ingredients are GL-V9 and a chemotherapy drug; the chemotherapy drug is the PARP1 inhibitor Olaparib.

2. The pharmaceutical composition according to claim 1, characterized in that, The GL-V9 includes the GL-V9 compound or a pharmaceutically acceptable salt thereof.

3. The use of the pharmaceutical composition of claim 1 in the preparation of a drug for treating tumors, wherein the tumor is leukemia or lung cancer.

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