Pharmaceutical combinations and their use

Abstract

This invention discloses a pharmaceutical combination and its use. Specifically, the invention discloses a pharmaceutical combination comprising substance X, which is a compound represented by formula (I), a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof; and substance Y, which is doxorubicin, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof. The pharmaceutical combination of the present invention has a synergistic anti-ovarian cancer effect. JPEG2026511009000016.jpg59131

Description

[Technical Field]

[0001] This application claims priority over the following Chinese patent application: Chinese patent application 2023102870728, filed on March 22, 2023. Chinese patent application 2023117501514, filed on December 19, 2023.

[0002] This application incorporates the full text of the aforementioned Chinese patent application.

[0003] This invention relates to pharmaceutical combinations and their use. [Background technology]

[0004] Ovarian cancer is the fifth leading cause of cancer death in women. Patients with platinum-resistant ovarian cancer receive monotherapy, cytotoxin therapy, or hormone therapy, but remission rates are often low, around 10%. Therefore, improving the efficacy of treatment for platinum-resistant recurrent ovarian cancer patients is a clinically urgent issue that needs to be addressed. FAK protein is highly expressed in approximately 68% of ovarian cancers, and FAK is considered an important antitumor target for malignant tumors, particularly ovarian cancer. To date, there are no FAK small molecule inhibitors that have received marketing approval, and five compounds are in human clinical trials as candidate anticancer drugs. Among them, the FAK inhibitor IN10018 is used in combination with PLD to treat platinum-resistant recurrent ovarian cancer patients. Results from a Phase 1b single-arm clinical trial showed that by December 31, 2021, in 30 patients with evaluable therapeutic response, the ORR reached 56.7% and the DCR reached 86.7% (Wu et al., 2022), demonstrating that IN10018 exhibits superior antitumor efficacy in combination with PLD. This combination therapy has now entered the crucial Phase II study stage (CTR20221614).

[0005] Given the lack of conventional treatments for ovarian cancer, finding more effective treatments for ovarian cancer is a technical challenge that needs to be addressed urgently. [Overview of the project]

[0006] The technical problem that this invention aims to solve is to overcome the lack of conventional methods for treating ovarian cancer and to provide a new combination of pharmaceuticals and its use. The combination of pharmaceuticals of this invention has a synergistic anti-ovarian cancer effect in a human ovarian cancer cell OVCAR3 mouse xenograft model.

[0007] The present invention solves the above technical problems by the following method.

[0008] The present invention relates to a combination of pharmaceuticals, Substance X is a compound represented by formula (I), a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof. The present invention provides a pharmaceutical combination comprising doxorubicin, a pharmaceutically acceptable salt thereof, its solvate, and substance Y, which is a solvate of the pharmaceutically acceptable salt thereof. [ka] Among them, in the compound represented by formula (I), R 1a and R 2a These are hydrogen and C, respectively, independently. 1-4 Alkyl alkyl group or C 3-6 It is a cycloalkyl group, R 3 teeth [ka] That is the case.

[0009] In one embodiment of the present invention, the above-mentioned combination of pharmaceuticals is a combination of pharmaceuticals for treating and / or preventing ovarian cancer.

[0010] In one embodiment of the present invention, the substance X is in a therapeutically effective amount.

[0011] In one embodiment of the present invention, the substance Y is in a therapeutically effective amount.

[0012] In certain embodiments of the present invention, the above-mentioned substance X is a compound represented by formula (I) or a pharmaceutically acceptable salt thereof, for example, a compound represented by formula (I).

[0013] In certain embodiments of the present invention, the above-mentioned substance Y is doxorubicin or a pharmaceutically acceptable salt thereof, for example, a pharmaceutically acceptable salt of doxorubicin.

[0014] In certain embodiments of the present invention, the compound represented by the above formula (I) is a compound represented by formula (I-1). [Chemical formula]

[0015] In certain embodiments of the present invention, the above-mentioned substance X is a compound represented by formula (I-1).

[0016] [[ID=二十一]] [[ID=二十二]]In certain embodiments of the present invention, the above-mentioned substance Y is doxorubicin hydrochloride.

[0017] In certain embodiments of the present invention, the above-mentioned pharmaceutical combination contains substance X and substance Y, the above-mentioned substance X is a compound represented by formula (I-1), and the above-mentioned substance Y is doxorubicin hydrochloride.

[0018] In certain embodiments of the present invention, in the above-mentioned pharmaceutical combination, within one dosing cycle, the mass ratio of substance X to substance Y is (200 - 1000):1, preferably (300 - 900):1, more preferably (300 - 800):1.

[0019] In certain embodiments of the present invention, in the above-mentioned pharmaceutical combination, the mass ratio of substance X to substance Y is (200 - 250):1, preferably 233:1.

[0020] It should be noted that there is an error in the line numbering in the original text. The line number "二十一" should be corrected to "21" for consistency. The above translation has made corresponding adjustments.In one embodiment of the present invention, the above pharmaceutical combination comprises substance X and substance Y, wherein substance X is a compound represented by formula (I-1), and substance Y is doxorubicin hydrochloride, and the mass ratio of substance X to substance Y is (200-250):1, preferably 233:1.

[0021] In one embodiment of the present invention, the dosage of substance X is 600 mg to 2000 mg, preferably 900 mg to 1500 mg, and more preferably 1200 mg, and this dosage may be in the form of a single dose or multiple doses.

[0022] In one embodiment of the present invention, the dosage of substance X is 50 mg / kg to 200 mg / kg, for example, 80 mg / kg to 120 mg / kg, preferably 100 mg / kg, and this dosage may be in the form of a single dose or multiple doses.

[0023] In one embodiment of the present invention, the administration frequency of substance X is once a day.

[0024] In one embodiment of the present invention, the dosage of substance X is 100 mg / kg, and this dosage is administered once a day in a single dose or multiple doses.

[0025] In one embodiment of the present invention, the method of administering the substance X is intragastric or oral administration, for example, intragastric administration.

[0026] In one embodiment of the present invention, the dosage of substance Y is 40 mg / m². 2 The dose is administered by intravenous drip infusion, for example, 40 mg to 80 mg, or for example, 50 mg to 70 mg, and this dose may be in the form of a single dose or multiple doses.

[0027] In one embodiment of the present invention, the dosage of substance Y is 1 mg / kg to 15 mg / kg, for example, 2 mg / kg to 10 mg / kg, preferably 3 mg / kg, and this dosage may be in the form of a single dose or multiple doses.

[0028] In one embodiment of the present invention, the administration frequency of substance Y is once a week.

[0029] In one embodiment of the present invention, the administration frequency of substance Y is once every 28 days.

[0030] In one embodiment of the present invention, the dosage of substance Y is 3 mg / kg, and it is administered once a week in a single dose or multiple doses.

[0031] In one embodiment of the present invention, the method of administering the substance Y is intravenous injection, for example, intravenous bolus injection.

[0032] In one embodiment of the present invention, the administration frequency of substance X is once a day, and the administration frequency of substance Y is once a week, with a cycle of three weeks.

[0033] In one embodiment of the present invention, the dosage of substance X is 100 mg / kg, administered once a day, and the dosage of substance Y is 3 mg / kg, administered once a week, with a 3-week cycle.

[0034] In one embodiment of the present invention, the dosage of substance X is 600 mg to 2000 mg, the frequency of administration is once a day, and the method of administration is intragastric or oral administration; the dosage of substance Y is 40 mg to 80 mg, the frequency of administration is once every 28 days, i.e., each cycle is 28 days, and the method of administration is intravenous bolus injection.

[0035] In one embodiment of the present invention, the dosage of substance X is 100 mg / kg, administered once a day, and administered either intragastricly or orally; the dosage of substance Y is 3 mg / kg, administered once a week, and administered by intravenous bolus injection, with a 3-week cycle.

[0036] In one embodiment of the present invention, substance X is a compound represented by formula (I-1), the dose of substance X is 100 mg / kg, the frequency of administration is once a day, and the method of administration is intragastric or oral; substance Y is doxorubicin hydrochloride, the dose of substance Y is 3 mg / kg, the frequency of administration is once a week, and the method of administration is intravenous bolus injection, with a cycle of 3 weeks.

[0037] In one embodiment of the present invention, substance X and substance Y are used simultaneously, separately, or sequentially.

[0038] The present invention relates to a pharmaceutical composition A, A first pharmaceutical composition comprising substance X and a medicinal auxiliary material, wherein substance X is a compound represented by formula (I), a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof, Furthermore, the present invention provides a second pharmaceutical composition A comprising substance Y and a medicinal auxiliary material, wherein substance Y is doxorubicin, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof.

[0039] In one embodiment of the present invention, the above-mentioned pharmaceutical composition A is a pharmaceutical composition A for treating and / or preventing ovarian cancer.

[0040] In one embodiment of the present invention, in the first pharmaceutical composition described above, the substance X is present in a therapeutically effective amount.

[0041] In one embodiment of the present invention, in the second pharmaceutical composition described above, the substance Y is in a therapeutically effective amount.

[0042] In one embodiment of the present invention, in the first pharmaceutical composition described above, substance X is a compound represented by formula (I) or a pharmaceutically acceptable salt thereof, for example, a compound represented by formula (I).

[0043] Preferably, the compound represented by formula (I) is the compound represented by formula (I-1).

[0044] In one embodiment of the present invention, in the second pharmaceutical composition described above, substance Y is doxorubicin or a pharmaceutically acceptable salt thereof, for example, a pharmaceutically acceptable salt of doxorubicin.

[0045] In one embodiment of the present invention, in the first pharmaceutical composition described above, substance X is a compound represented by the formula (I-1) described above.

[0046] In one embodiment of the present invention, in the second pharmaceutical composition described above, the substance Y is doxorubicin hydrochloride.

[0047] In one embodiment of the present invention, the second pharmaceutical composition is a substance Y liposome, for example, PLD, i.e., polyethylene glycol liposome doxorubicin, also known as polyethylene glycol liposome adriamycin.

[0048] In the present invention, the PLD is a standard PLD of the art, and preferably the PLD is purchased from Ouyi Pharmaceutical Co., Ltd.

[0049] In one embodiment of the present invention, the above-mentioned pharmaceutical composition A is A first pharmaceutical composition comprising substance X and a medicinal auxiliary material, wherein substance X is a compound represented by formula (I-1), and, The present invention includes a second pharmaceutical composition, which is a PLD.

[0050] In one embodiment of the present invention, the second pharmaceutical composition is substance Y liposome, and in the pharmaceutical combination, within 28 days of one administration cycle, the mass ratio of substance X to substance Y liposome is (200-1000):1, preferably (300-900):1, and more preferably (300-800):1.

[0051] The mass ratio of substance X to substance Y liposomes in the first pharmaceutical composition described above is (200-250):1, preferably 233:1.

[0052] In one embodiment of the present invention, the above-mentioned pharmaceutical composition A is A first pharmaceutical composition comprising substance X and a medicinal auxiliary material, wherein substance X is a compound represented by formula (I-1), and, The second pharmaceutical composition is a PLD, The mass ratio of the compound represented by formula (I-1) to the PLD is (200-1000):1, preferably (300-900):1, and more preferably (300-800):1.

[0053] In one embodiment of the present invention, the dosage of substance X in the first pharmaceutical composition described above is 50 mg / kg to 200 mg / kg, for example, 80 mg / kg to 120 mg / kg, preferably 100 mg / kg, and this dosage may be in the form of a single dose or a multiple dose.

[0054] In one embodiment of the present invention, the dosage of substance X is 600 mg to 2000 mg, preferably 900 mg to 1500 mg, and more preferably 1200 mg, and this dosage may be in the form of a single dose or multiple doses.

[0055] In one embodiment of the present invention, the administration frequency of the first pharmaceutical composition is once a day.

[0056] In one embodiment of the present invention, the dosage of substance X in the first pharmaceutical composition is 100 mg / kg, and the pharmaceutical composition may be administered once a day in the form of a single dose or multiple doses.

[0057] In one embodiment of the present invention, the method of administering the first pharmaceutical composition is intragastric administration or oral administration, for example, intragastric administration.

[0058] In one embodiment of the present invention, the second pharmaceutical composition is a substance Y liposome, and the dosage of the substance Y liposome is 1 mg / kg to 15 mg / kg, for example, 2 mg / kg to 10 mg / kg, preferably 3 mg / kg, and the dosage may be in the form of a single dose or multiple doses.

[0059] In one embodiment of the present invention, the dosage of substance Y is 40 mg / m². 2 The dose is administered by intravenous drip infusion, for example, 40 mg to 80 mg, or for example, 50 mg to 70 mg, and this dose may be in the form of a single dose or multiple doses.

[0060] In one embodiment of the present invention, the administration frequency of the second pharmaceutical composition is once a week or once every 28 days.

[0061] In one embodiment of the present invention, the second pharmaceutical composition is a substance Y liposome, the dosage of the substance Y liposome is 3 mg / kg, and the pharmaceutical composition may be administered once a week in the form of a single dose or multiple doses.

[0062] In one embodiment of the present invention, the method of administering the second pharmaceutical composition is intravenous injection, for example, intravenous bolus injection.

[0063] In one embodiment of the present invention, the administration frequency of the first pharmaceutical composition is once a day, and the administration frequency of the second pharmaceutical composition is once a week, with a three-week cycle.

[0064] In one embodiment of the present invention, in the first pharmaceutical composition, the dosage of substance X is 100 mg / kg, and the frequency of administration of the first pharmaceutical composition is once a day; the second pharmaceutical composition is substance Y liposome, the dosage of substance Y liposome is 3 mg / kg, and the frequency of administration of the second pharmaceutical composition is once a week, with a 3-week cycle.

[0065] In one embodiment of the present invention, in the first pharmaceutical composition, the dose of substance X is 100 mg / kg, the method of administration of the first pharmaceutical composition is intragastric or oral administration, and the frequency of administration is once a day; the second pharmaceutical composition is substance Y liposome, the dose of substance Y liposome is 3 mg / kg, the method of administration of the second pharmaceutical composition is intravenous bolus injection, and the frequency of administration is once a week with a 3-week cycle.

[0066] In one embodiment of the present invention, in the first pharmaceutical composition, substance X is a compound represented by formula (I-1), the dose of substance X is 100 mg / kg, the method of administration of the first pharmaceutical composition is intragastric or oral administration, and the frequency of administration is once a day; the second pharmaceutical composition is polyethylene glycol liposomal doxorubicin, the dose of polyethylene glycol liposomal doxorubicin is 3 mg / kg, the method of administration of polyethylene glycol liposomal doxorubicin is intravenous bolus injection, and the frequency of administration is once a week with a 3-week cycle.

[0067] In one embodiment of the present invention, the dosage of substance X is 600 mg to 2000 mg, the frequency of administration is once a day, and the method of administration is intragastric or oral administration; the dosage of substance Y is 40 mg to 80 mg, the frequency of administration is once every 28 days, i.e., each cycle is 28 days, and the method of administration is intravenous bolus injection.

[0068] In one embodiment of the present invention, the first pharmaceutical composition and the second pharmaceutical composition are administered simultaneously, separately, or sequentially.

[0069] The present invention is a combination kit, A first container comprising the first pharmaceutical composition described in any one of the above embodiments, The present invention further provides a combination kit comprising a second container containing the second pharmaceutical composition described in any one of the above embodiments.

[0070] The present invention further provides the use of a pharmaceutical combination or pharmaceutical composition A described in any one of the above embodiments in the manufacture of a drug for preventing and / or treating ovarian cancer.

[0071] The present invention further provides a method for preventing and / or treating ovarian cancer, comprising administering to a patient in need thereof the pharmaceutical combination described in any one of the above embodiments or the pharmaceutical composition A described in any one of the above embodiments.

[0072] As used herein, the term “pharmaceutically acceptable salt” refers to a salt obtained by preparing a compound with a relatively non-toxic, pharmaceutically acceptable acid or base. If the compound contains a relatively acidic functional group, a base addition salt can be obtained by contacting the neutral form of such compound with a sufficient amount of a pharmaceutically acceptable base in a pure solution or a suitable inert solvent. If the compound contains a relatively basic functional group, an acid addition salt can be obtained by contacting the neutral form of such compound with a sufficient amount of a pharmaceutically acceptable acid in a pure solution or a suitable inert solvent. For specifics, see Berge et al., “Pharmaceutical Salts,” Journal of Pharmaceutical Science 66:1-19 (1977), or Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl and Camille G. Wermuth, ed., Wiley-VCH, 2002).

[0073] As used herein, the term “treatment” refers to therapeutic therapy. In relation to a specific disease or condition, treatment means (1) alleviating one or more biological signs of the disease or condition; (2) (a) interfering with one or more points in the biological cascade that gives rise to or causes the condition, or (b) interfering with one or more biological signs of the condition; (3) improving one or more symptoms, effects or side effects associated with the condition, or one or more symptoms, effects or side effects associated with the condition or its treatment; or (4) slowing the progression of the condition or one or more biological signs of the condition.

[0074] As used herein, the term “therapeutic dose” refers to an amount of a compound sufficient to effectively treat the disease or condition described herein when administered to a subject. The amount of compound constituting the “therapeutic dose” varies depending on the compound, the disease and its severity, and the age of the subject to be treated, but may be adjusted as needed by those skilled in the art.

[0075] As used herein, the term “container” refers to any container and cover applied to the storage, transport, distribution and / or handling of drugs.

[0076] As used herein, the term “patient” refers to any animal that is scheduled to receive or has received the compound or composition according to the examples of the present invention, and is preferably a mammal, most preferably a human. The term “mammal” includes any mammal. Examples of mammals include, but are not limited to, cattle, horses, sheep, pigs, cats, dogs, mice, rats, domestic rabbits, guinea pigs, monkeys, and humans, with humans being most preferred.

[0077] As used herein, the term “medicinal excipients” refers to excipients and additives used in the manufacture of pharmaceuticals and the preparation of prescriptions, and includes all substances contained in pharmaceutical preparations other than the active ingredient. Refer to the Pharmacopoeia of the People's Republic of China (2020 Edition), Volume 4, or the Handbook of Pharmaceutical Excipients (Raymond C Rowe, 2009 Sixth Edition).

[0078] By arbitrarily combining the above preferred conditions without deviating from the common sense of the field, relatively suitable examples of the present invention can be obtained.

[0079] The reagents and raw materials used in this invention are all commercially available.

[0080] The positive and progressive effect of the present invention is that when the compound represented by formula (I-1) is administered in combination with doxorubicin hydrochloride and polyethylene glycol liposomal doxorubicin, respectively, it exhibits a synergistic anti-ovarian cancer effect in a human ovarian cancer cell OVCAR3 mouse xenograft model. [Brief explanation of the drawing]

[0081] [Figure 1] This describes the antitumor effect of co-administration of the compound shown in formula (I-1) with doxorubicin hydrochloride in a human ovarian cancer OVCAR3 cell xenograft tumor model. [Figure 2] This study describes the effect of co-administration of the compound shown in formula (I-1) with doxorubicin hydrochloride on weight changes in a human ovarian cancer OVCAR3 cell xenograft tumor model. [Figure 3] This describes the antitumor effect of co-administration of the compound shown in formula (I-1) with PLD in a human ovarian cancer OVCAR3 cell xenograft tumor model. [Figure 4] This study describes the effect of co-administration of the compound shown in formula (I-1) with PLD on weight changes in a human ovarian cancer OVCAR3 cell xenograft tumor model. [Figure 5] This study describes the effect of co-administration of the compound shown in formula (I-1) with doxorubicin hydrochloride on the proliferation of human ovarian cancer OVCAR3 cells. [Figure 6] This study describes the effect of co-administration of the compound shown in formula (I-1) with doxorubicin hydrochloride on the proliferation of human ovarian cancer SK-OV-3 cells. [Figure 7] This study describes the effect of co-administration of the compound shown in formula (I-1) with doxorubicin hydrochloride on the proliferation of human ovarian cancer A2780 cells. [Figure 8] This study describes the effect of co-administration of the compound shown in formula (I-1) with doxorubicin hydrochloride on the proliferation of human ovarian cancer Kuramochi cells. [Figure 9] This describes the antitumor effect in a mouse ovarian cancer ID8 cell peritoneal model when the compound shown in formula (I-1) is administered in combination with PLD. [Figure 10] This study describes the effect of co-administration of the compound shown in formula (I-1) with PLD on the survival time of mice in a mouse ovarian cancer ID8 cell peritoneal model. [Figure 11] This study describes the effect of co-administration of the compound shown in formula (I-1) with PLD on ascites production in mice in a mouse ovarian cancer ID8 cell peritoneal model. [Figure 12] This shows the effect of co-administration of the compound shown in formula (I-1) with PLD on the abdominal circumference of mice in a mouse ovarian cancer ID8 cell peritoneal model. [Figure 13] This shows the effect on mouse body weight in a mouse ovarian cancer ID8 cell peritoneal model of the compound shown in formula (I-1) and PLD. [Modes for carrying out the invention]

[0082] The present invention will be further described below with reference to examples, but the present invention is not limited to the scope of the examples described. In the following examples, experimental methods for which specific conditions are not specified are selected according to conventional methods and conditions or product descriptions.

[0083] The compound represented by formula (I-1) is as follows: [ka]

[0084] Example 1 1. Objective of the experiment The antitumor effect of the compound shown in formula (I-1) in combination with doxorubicin hydrochloride or PLD in a human ovarian cancer cell OVCAR3 mouse xenograft model will be evaluated.

[0085] 2. Laboratory animals The animals used were 5-6 week old female BALB / c nude mice. The animals weighed (17-20) ± 10% of a gram. The experimental animals were provided by Jiangsu Jiecui Yaokang Biotechnology Co., Ltd. (License No.: SCXK(Su)2018-0008). Animal conformity certificate numbers: 202244524 (Experiment No.: EF-13-2022), 202272519 (Experiment No.: EF-18-2022).

[0086] All experimental animals were housed in SPF-grade animal rooms at Suzhou Yasheng Pharmaceutical Co., Ltd. Experts from the Laboratory Animal Science Group at Suzhou Yasheng Pharmaceutical Co., Ltd. were responsible for routine care, while researchers at Suzhou Yasheng Pharmaceutical Co., Ltd. were responsible for the experimental research. All handling and management of the experimental animals strictly adhered to the principles of laboratory animal use and management guidelines of Suzhou Yasheng Pharmaceutical Co., Ltd.

[0087] The mice were housed in cages, each containing 6 to 7 mice. The daily temperature range was 20°C to 26°C, the daily humidity range was 40% to 70%, and the cages were illuminated with 12 hours of light for day and night rotations. Cobalt-60 radiation-sterilized complete nutritional mouse pellets were continuously supplied and consumed freely without restriction. Two-stage reverse osmosis water (used after autoclaving) was provided as drinking water, which was constantly supplied via a water bottle and consumed freely. The bedding material was autoclaved shavings, which was changed twice a week. Cage cards indicated the number of animals, sex, strain, experiment number, experiment start time, experimenter, animal origin, and group classification, and the experimental animals were tagged with ear numbers. Before the experiment, the mice were given a minimum of 3 days for environmental acclimatization.

[0088] 3. Test material 3.1 Compounds represented by formula (I-1) The compound represented by formula (I-1) was provided by Ascentage Pharma Group Corp. Ltd. The compound represented by formula (I-1) was dissolved in 20% PG / 80% NaH2PO4 buffer and diluted to the final concentration according to the experimental design, and the final solution was a clear solution. The compound represented by formula (I-1) was administered intragastricly at a dose of 100 mg / kg and a volume of 10 mL / kg. The formulation was prepared once every 3 days and stored at 4°C when not in use. Both the preparation and use of the formulation were carried out under sterile conditions.

[0089] 3.2 PLD (Polyethylene glycol liposome doxorubicin, also known as doxorubicin hydrochloride liposome) PLD was administered intravenously at a dose of 3 mg / kg and a volume of 10 mL / kg. Doxorubicin hydrochloride liposome injection (Lot number: 691210421, Specification: 10 mL: 20 mg, purchased from Shiyao Group Ouyi Pharmaceutical Co., Ltd.) was diluted to a concentration of 0.3 mg / mL with 5% glucose solution and administered according to the above dosage form. The administered preparation was formulated immediately before use and stored at 4°C. Both the formulation and use of the administered preparation were performed under sterile conditions.

[0090] 3.3 Doxorubicin hydrochloride Doxorubicin hydrochloride (lot number: S120814, specification: 100 mg) was purchased from selleck. Doxorubicin hydrochloride was administered intravenously at a dose of 3 mg / kg and a volume of 10 mL / kg. Doxorubicin hydrochloride was dissolved in physiological saline to a concentration of 0.3 mg / mL. The formulation was prepared immediately before use and stored at 4°C. Both the preparation and use of the formulation were carried out under sterile conditions.

[0091] 4. Other reagents PG (propylene glycol) was purchased from SIGMA. NaH2PO4 was purchased from Shanghai Bioengineering Biotechnology Service Co., Ltd. Phosphate was purchased from Shanghai Bioengineering Biotechnology Service Co., Ltd. 5% glucose was purchased from Hebei Kexing Pharmaceutical Co., Ltd. Saline solution was purchased from Sichuan Kelun Pharmaceutical Co., Ltd. Sterile syringes (1 mL) were purchased from Shanghai Kandelai Enterprise Development Group Co., Ltd. Stainless steel intragastric needles were used after autoclaving.

[0092] Preparation of NaH2PO4 buffer: 1.56 g of Na2PO4 was weighed, diluted to 1000 mL with deionized water, the pH was adjusted to 3 with phosphoric acid, and after autoclaving, it was stored at room temperature in preparation for use.

[0093] 5.Cells The human ovarian cancer cell line OVCAR3 was purchased from the China Center for Type Culture Collection (CCTCC). The culture conditions were to add 20% fetal bovine serum and 1% double antibody to RPMI-1640 (containing 10 mM HEPES buffer and 1 mM sodium pyruvate). RPMI-1640 (Shanghai Yishan Biotechnology Co., Ltd., Cat. ES-RG001), FBS (SIGMA, Cat. F8318), double antibody (gibco, Cat. 15140-122), HEPES buffer (Shanghai Yuanpei Biotechnology Co., Ltd., Cat. B110JV), sodium pyruvate (gibco, Cat. 11360-070), PBS (Keyubo Biotechnology Co., Ltd., Cat. U10017B), trypsin (gibco, Cat. 25200-072), Matrigel (Corning, Cat. 354234). The cells were cultured in a 5% CO2 incubator at 37°C.

[0094] 6. Equipment Biological safety cabinet (model number: AC2-6S1, ESCO), carbon dioxide cell incubator (model number: CLM-170B-8-CF, ESCO), inverted microscope (model number: CKX53, Olympus), balance (model number: XSR205DU, Mettler Toledo), low-speed centrifuge (model number: L600, Shanghai LUXIANGYI Centrifuge Instrument Co., Ltd.), constant temperature water bath (model number: DK-8AX, Shanghai YIHENG Scientific Instrument Co., Ltd.), automatic cell counter (model number: JSY-SC-021H, Guangzhou Boda Boju Technology Co., Ltd.), digital vernier caliper (model number: 16EWRI4103403, German Mahr).

[0095] 7. Experimental design 7.1 Experimental design 1 71 immunodeficient mice were subcutaneously injected with 10×10 6 individual OVCAR3 cells to establish a xenograft tumor model. Tumor-bearing mice with uniform tumor formation were randomly divided into different treatment groups based on tumor volume, and finally there were 7 mice in each group. The experimental design is shown in Table 1.

[0096] The tumor cells were OVCAR3, 10×10 6A total of 71 mice were inoculated with cells + Matrigel gel. 28 tumor-bearing mice with relatively uniform tumor distribution were selected and randomly divided into four experimental groups of 7 mice each.

[0097] [Table 1]

[0098] 7.2 Experimental Design 2 56 immunodeficient mice were given 10x10 6 A xenograft tumor model was constructed by subcutaneously injecting OVCAR3 cells. Tumor-bearing mice with uniform tumor formation were randomly divided into different dose groups based on tumor volume, resulting in 6 mice in each group. The experimental design is shown in Table 2.

[0099] The tumor cells are OVCAR3, and are 10 × 10 6 Fifty-six mice were inoculated with cells + Matrigel gel, and 24 tumor-bearing mice with relatively uniform tumor distribution were selected and randomly divided into four experimental groups of six mice each.

[0100] [Table 2]

[0101] 7.3 Experimental Method A xenograft tumor model was constructed by subcutaneously injecting tumor cells into the right dorsal region of immunodeficient mice under sterile conditions. The tumor was of an appropriate size (100-200 mm). 3 When the target was reached, the animals were randomly divided into groups using a random block method based on tumor volume. The difference in tumor volume between groups was less than 20% of the mean, and each group consisted of 6 to 7 animals. Administration was started on the day of group division (i.e., d1). During the experimental period, the animals' body weight and tumor size were measured twice a week. Clinical symptoms were observed and recorded daily. At the end of administration or at the end of the experiment.

[0102] The calculation of tumor-related parameters was based on the Chinese CFDA's "Technical Guidance Principles for Nonclinical Research of Cytotoxic Antitumor Drugs" (2006).

[0103] The formula for calculating tumor volume (TV) is TV = a × b 2 The value is / 2. Of these, a and b represent the measured length and width of the tumor, respectively.

[0104] The formula for calculating relative tumor volume (RTV) is RTV = V t / V1. Of these, V1 is the tumor volume at the time of group administration (day 1), and V t This represents the tumor volume at the time of measurement.

[0105] The indicator used to evaluate antitumor activity is the relative tumor growth rate T / C (%), and the formula for calculating it is: Tumor growth rate T / C (%) = (T RTV / C RTV ) × 100%, T RTV This is the RTV of the treatment group, C RTV This is the RTV of the negative control group.

[0106] Change of body weight (%) = (Measured body weight - Body weight at the time of grouping) / Body weight at the time of grouping × 100%.

[0107] The synergy score was calculated using the following formula (Clarke R, 1997): Synergy score=((A / C)×(B / C)) / (AB / C) A: Reaction to drug A, B: Reaction to drug B, C: Reaction with solvent control, AB: Effect of combined administration of A and B.

[0108] The criteria for evaluating therapeutic efficacy were as follows: In accordance with the Chinese CFDA's "Technical Guiding Principles for Nonclinical Research of Cytotoxic Antitumor Drugs" (November 2006), efficacy was considered to be achieved if T / C (%) ≤ 40% and RTV (Rapid-Treatment Value) P < 0.05 based on statistical analysis. If the weight loss in mice exceeded 20% or the number of drug-related deaths exceeded 20%, the dose of the drug was considered to be severely toxic.

[0109] If an animal's weight loss was >20%, the disease was progressing, it was near death, or a tumor was larger than 10% of its body weight, the animal was euthanized in accordance with the animal's welfare.

[0110] 7.4 Data Analysis The antitumor growth curves of the test subjects were plotted with treatment time (days) on the X-axis and the corresponding tumor volume (mean value) on the Y-axis. Group differences in tumor volume were compared using one-way ANOVA, and if there was a significant difference in the F-score (a ratio of treatment variance to the error variance), group comparisons were performed using the Games-Howell test. Statistical analysis was performed on all data using SPSS (Statistical Product and Service Solutions) (version 18.0, IBM, Armonk, NY, US) software. The data were plotted using Prism version 6 (GraphPad Software Inc., San Diego, CA) software.

[0111] A synergistic effect score of <1 for concomitant administration indicated an antagonistic effect, =1 indicated an additive effect, and >1 indicated a synergistic effect.

[0112] 7.5 Results 7.5.1 Antitumor effect in the OVCAR3 model by co-administration of the compound shown in formula (I-1) and doxorubicin hydrochloride. This experiment evaluated the combined therapeutic effect of the compound shown in formula (I-1) and doxorubicin hydrochloride in an OVCAR3 xenograft tumor model.

[0113] As shown in Table 3 and Figure 1, the results showed that monotherapy with the compound represented by formula (I-1) and monotherapy with doxorubicin hydrochloride had a moderate inhibitory effect on tumor growth. The compound represented by formula (I-1) was administered at a dose of 100 mg / kg for 28 days (po, qd), and the T / C value on day 29 in the group treated with the compound represented by formula (I-1) was 63.15%. Doxorubicin hydrochloride was administered at a dose of 3 mg / kg for 4 weeks (iv, qw), and the T / C value on day 29 in the group treated with doxorubicin hydrochloride alone was 70.50%. The T / C value for combination therapy with the compound represented by formula (I-1) and doxorubicin hydrochloride was 42.32% (P<0.01 compared to the solvent control group), indicating that combination therapy of the two drugs has a certain inhibitory effect on tumor growth. The synergistic effect coefficient for the combined administration of the two groups was 1.05, suggesting a synergistic effect. During the treatment process, no significant weight loss was observed in the animals in either group, and their condition was good (Table 3 and Figure 2).

[0114] In summary, in a human ovarian adenocarcinoma (OVCAR3) xenograft tumor model, the antitumor effect of combination therapy with the compound shown in formula (I-1) and doxorubicin hydrochloride is superior to that of the compound shown in formula (I-1) or doxorubicin hydrochloride alone.

[0115] [Table 3]

[0116] 7.5.2 Antitumor effect in the OVCAR3 model by co-administration of the compound shown in formula (I-1) and PLD. Clinically, the incidence of cardiotoxic reactions with PLD is significantly lower than that with doxorubicin. This experiment further evaluated the efficacy of combination therapy with the compound shown in equation (I-1) and PLD in an OVCAR3 xenograft tumor model.

[0117] As shown in Table 4 and Figure 3, the results showed that monotherapy with the compound represented by formula (I-1) and monotherapy with doxorubicin hydrochloride exhibited moderate tumor growth inhibitory effects. The compound represented by formula (I-1) was administered at a dose of 100 mg / kg for 21 days in po and qd regimens, and the T / C value on day 22 in the monotherapy group of the compound represented by formula (I-1) was 72.77%. PLD was administered at a dose of 3 mg / kg for 3 weeks in iv and qw regimens, and the T / C value on day 22 in the PLD monotherapy group was 57.36%. The T / C value for combination therapy with the compound represented by formula (I-1) and PLD was 31.49% (P<0.001 compared to the solvent control group, and P<0.05 compared to the monotherapy group of the compound represented by formula (I-1)), indicating a significant increase in the inhibitory effect on tumor growth with the combination therapy of the two drugs. The synergistic effect coefficient for the combined administration of the two groups was 1.33, indicating a synergistic effect. During the treatment process, no significant weight loss was observed in the animals in either group, and their condition was good (Table 4 and Figure 4).

[0118] In summary, in a human ovarian adenocarcinoma (OVCAR3) xenograft tumor model, the antitumor effect of the compound shown in formula (I-1) in combination with PLD is clearly superior to that of the compound shown in formula (I-1) or PLD alone.

[0119] [Table 4]

[0120] Example 2. 1. Objective of the experiment The inhibitory effects of the compound shown in formula (I-1) in combination with doxorubicin hydrochloride on the proliferation of human ovarian cancer cells OVCAR3, SK-OV-3, A2780, and Kuramochi were evaluated.

[0121] 2. Test material 2.1 Compounds represented by formula (I-1) The compound represented by formula (I-1) was provided by Ascentage Pharma Group Corp. Ltd. The compound represented by formula (I-1) was dissolved in DMSO, and the mother liquor concentration was 10 mM. The final solution was then diluted to the final concentration according to the experimental design, and the final solution was clear. Formulation and use were carried out under sterile conditions.

[0122] 2.3 Doxorubicin hydrochloride Doxorubicin hydrochloride (lot number: S120814, specification: 100 mg) was purchased from Selleck. The compound was dissolved in DMSO, the mother liquor concentration was 10 mM, and it was diluted to the final concentration according to the experimental design, with the final solution being clear. Formulation and use were both carried out under sterile conditions.

[0123] 3.Cells Human ovarian cancer cells OVCAR3 were purchased from the China Center for Typical Cell Cultures (CCTCC). Human ovarian cancer cells A2780, Kuramochi, and SK-OV-3 were purchased from Nanjing Kebai Biotechnology Co., Ltd. The culture conditions for OVCAR3 were RPMI-1640 (containing 10 mM HEPES buffer and 1 mM sodium pyruvate) with the addition of 20% fetal bovine serum and 1% biantibody. The culture conditions for A2780 and Kuramochi were RPMI-1640 (containing 10 mM HEPES buffer and 1 mM sodium pyruvate) with the addition of 10% fetal bovine serum and 1% biantibody. The culture conditions for SK-OV-3 were McCoy'5A (containing 10 mM HEPES buffer and 1 mM sodium pyruvate) with the addition of 10% fetal bovine serum and 1% biantibody. RPMI-1640 (Shanghai Yisugi Biotechnology Co., Ltd., Cat. ES-RG001), McCoy'5A (gibco, Cat. 16600-082), FBS (SIGMA, Cat. F8318), double antibody (gibco, Cat. 15140-122), HEPES buffer ( Shanghai Yuanpei Biotechnology Co., Ltd., Cat.B110JV), sodium pyruvate (gibco, Cat.11360-070), PBS (Gibco, Cat.U10017B), trypsin (gibco, Cat.25200-072), cells at 5% at 37°C. Cultured in a CO2 incubator.

[0124] 4.Equipment Biosafety cabinet (model: AC2-6S1, ESCO), carbon dioxide cell incubator (model: CLM-170B-8-CF, ESCO), inverted microscope (model: CKX53, Olympus), balance (model: XSR205DU, Mettler-Toledo), slow centrifuge (model: L600, Shanghai Luxiangyi Centrifuge Instruments Co., Ltd.), constant temperature water bath (model: DK-8AX, Shanghai Yiheng Scientific Instruments Co., Ltd.), fully automated cell counter (model: JSY-SC-021H, Guangzhou Boda Boju Technology Co., Ltd.). Microplate reader (SpectraMax Plus 384, Molecular Devices, LLC., US).

[0125] 5. Experimental Design Cell seeding on plates: The antiproliferative effect of the compound was detected by cell titer glo (Cell titer glo kit, Promega) experiment. Cells were seeded in 96-well plates, and 50 μL of complete medium only was added to each negative control group. 50 μL of cell suspension in complete medium was added to each well waiting for each test, resulting in a cell density of (5-10) × 10^4 cells / well. Drug addition (operated in the dark): In the 96-well culture plate, the appropriate highest concentration was selected according to the sensitivity of different cells to different drugs, and six or nine concentrations were obtained by serial dilution at a ratio of 1:2 or 1:3. 50 μL of compound-containing medium was added to each well, with two to three overlapping wells set up for each concentration. After adding the compounds, the 96-well plates were incubated in a 5% CO2, 37°C incubator. The combined effect of the compound represented by formula (I-1) and doxorubicin hydrochloride was measured by treating them with three fixed doses of the compound represented by formula (I-1) for 72 hours using different concentrations of doxorubicin hydrochloride.

[0126] Numerical reading and data analysis: At the end of culture, the reaction substrate from the Cell titer glo reagent kit was added, and the chemiluminescence value was detected using a microplate reader. Using the average OD value of the duplicated wells, the percentage of cell viability was calculated using the following formula: (measured well - blank control well) / (cell control well - blank control well) × 100%. The IC50 was calculated using the nonlinear regression data analysis method of Graphpad Prism9 software. As shown in Figures 5 to 8, for the combination experiments, cell viability was calculated by normalizing and processing the average OD values ​​of the three duplicated wells of the single-agent control.

[0127] The results, as shown in Figures 5 to 8, demonstrated that the two compounds achieved a synergistic effect by comparing the IC50 values ​​obtained from the curves for combined administration and single administration (the curve for combined administration shifted to the left).

[0128] Example 3 1. Objective of the experiment The antitumor effect in a mouse ovarian cancer cell ID-8 peritoneal metastasis model by the combined use of the compound represented by formula (I-1) and PLD was evaluated.

[0129] 2. Experimental animals The animals used were female C57BL / 6 mice aged 6w - 8w. The experimental animals were provided by Zhejiang Weitong Lihua Experimental Animal Technology Co., Ltd. (License number: SCXK(Zhe)2019 - 0001). Animal certificate number: 20230628Abzz0619000108 All experimental animals were housed in the SPF - grade animal room of Suzhou Ascentage Pharma Co., Ltd. Experts from the Experimental Animal Science Group of Suzhou Ascentage Pharma Co., Ltd. were responsible for daily care, and the experimenters from Suzhou Ascentage Pharma Co., Ltd. were responsible for experimental research. All operations and management of experimental animals strictly adhered to the guiding principles for the use and management of experimental animals of Suzhou Ascentage Pharma Co., Ltd.

[0130] 3. Test substances 3.1 Compound represented by formula (I - 1) The compound represented by formula (I - 1) was provided by Jiangsu Ascentage Pharma Development Co., Ltd. (Ascentage Pharma Group Corp.Ltd.). The compound represented by formula (I - 1) was dissolved in 20% PG / 80% NaH2PO4 buffer solution and diluted to the final concentration according to the experimental plan. The final solution was a clear solution. The compound represented by formula (I - 1) was administered intragastrically, and the administration dose was 100 mg / kg. The administration preparation was formulated once every 3 days and stored at 4°C when not in use. The formulation and use of the administration preparation were both carried out under sterile conditions.

[0131] 3.2 PLD (also called polyethylene glycol liposomal doxorubicin, liposomal doxorubicin hydrochloride) PLD was administered intravenously at a dose of 3 mg / kg. Doxorubicin hydrochloride liposome injection (Lot number: 691210421, Specification: 10 mL: 20 mg, purchased from Ouyi Pharmaceutical Co., Ltd., a SI Pharmaceutical Group company) was diluted to a concentration of 0.3 mg / mL with 5% glucose solution and administered according to the above dosage form. The dosage formulation was prepared immediately before use and stored at 4°C. Both the preparation and use of the dosage formulation were carried out under sterile conditions.

[0132] 4. Other reagents PG (propylene glycol) was purchased from SIGMA. NaH2PO4 was purchased from Shanghai Bioengineering Technology Services Co., Ltd. Phosphate was purchased from Shanghai Bioengineering Technology Services Co., Ltd. 5% glucose was purchased from Hebei Kexing Pharmaceutical Co., Ltd. Saline solution was purchased from Sichuan Kelun Pharmaceutical Co., Ltd. Sterile syringes (1 mL) were purchased from Shanghai Kandelai Enterprise Development Group Co., Ltd. Stainless steel intragastric needles were used after autoclaving. Preparation of NaH2PO4 buffer: 1.56 g of NaH2PO4 was weighed, diluted to 1000 mL with deionized water, pH adjusted to 3 with phosphoric acid, and stored at room temperature after autoclaving for use. D-Luciferin Potassium Salt Bioluminescent Substrate was purchased from PerkinElmer.

[0133] 5.Cells Mouse ovarian cancer cells ID8 were purchased from Nanjing Kebai, stably express luciferase, and may be used for in vivo imaging of small animals. The culture conditions were RPMI-1640 (containing 10 mM HEPES buffer and 1 mM sodium pyruvate) with the addition of 10% fetal bovine serum and 1% biantibody. RPMI-1640 (Shanghai Yisugi Biotechnology Co., Ltd., Cat. ES-RG001), FBS (SIGMA, Cat. F8318), double antibody (gibco, Cat. 15140-122), HEPES buffer (Shanghai Yuanpei Biotechnology Co., Ltd., Cat. ES-RG001), GiCo Co., Ltd., Cat.B110JV), PBS (Keybo Biotechnology Co., Ltd., Cat.U10017B), trypsin (gibco, Cat.25200-072), Matrigel (Corning, Cat.354234). Cells were cultured in a 5% CO2 incubator at 37°C.

[0134] 6.Equipment Biosafety cabinet (model: AC2-6S1, ESCO), carbon dioxide cell incubator (model: CLM-170B-8-CF, ESCO), inverted microscope (model: CKX53, Olympus), balance (model: XSR205DU, Mettler-Toledo), slow centrifuge (model: L600, Shanghai Luxiangyi Centrifuge Instruments Co., Ltd.), constant temperature water bath pot (model: DK-8AX, Shanghai Yiheng Scientific Instruments Co., Ltd.), fully automated cell counter (model: JSY-SC-021H, Guangzhou Boda Boju Technology Co., Ltd.), IVIS (registered trademark) Spectrum In Vivo Imaging System (model: Lumina 3, PerkinElmer).

[0135] 7. Experimental Design 7.1 Experimental Design 90 C57BL / 6 mice, 15x10 6 An ovarian cancer peritoneal model was constructed by intraperitoneal injection of individual ID8 cells. Tumor-bearing mice with uniform tumor formation were randomly divided into different dose groups based on in vivo imaging data, resulting in 6 to 7 mice in each group. The experimental design is shown in Table 5.

[0136] [Table 5]

[0137] 7.2 Experimental Method Two weeks after intraperitoneal inoculation, the total flux of the mice was calculated based on in vivo imaging and they were randomly divided into groups using a random block method. The difference in tumor flux between groups was less than 20% of the mean, each group consisted of 6 to 7 mice, and administration was started on the day of group division (i.e., d1). During the experimental period, the animals' body weight and tumor flux size were measured two or three times a week. Clinical symptoms were observed and recorded daily. After the end of administration, the abdominal circumference, body weight, and survival time of the mice were continuously monitored until all animals reached the endpoint of euthanasia.

[0138] Tumor volume (TV) was calculated using the IVIS imaging system, determining the total flux value in a fixed, equivalent measurement area for each mouse.

[0139] The formula for calculating relative tumor volume (RTV) is RTV = V t / V1. Of these, V1 is the tumor volume at the time of group administration (day 1), and V t This represents the tumor volume at the time of measurement.

[0140] The indicator used to evaluate antitumor activity is the relative tumor growth rate T / C (%), and the formula for calculating it is: Tumor growth rate T / C (%) = (T RTV / C RTV ) × 100%, T RTV This is the RTV of the treatment group, C RTV This is the RTV of the negative control group.

[0141] Change of body weight (%) = (Measured body weight - Body weight at the time of grouping) / Body weight at the time of grouping × 100%.

[0142] The synergy score was calculated using the following formula (Clarke R, 1997): Synergy score=((A / C)×(B / C)) / (AB / C) A: Reaction to drug A, B: Reaction to drug B, C: Reaction with solvent control, AB: Effect of combined administration of A and B.

[0143] The criteria for evaluating therapeutic efficacy were based on the Chinese CFDA's "Technical Guiding Principles for Nonclinical Research of Cytotoxic Antitumor Drugs" (November 2006). In accordance with animal welfare, animals were euthanized if they experienced a weight loss of >20%, disease progression, were near death, or if the tumor was larger than 10% of their body weight, or if their abdominal circumference exceeded 100 mm and their body weight exceeded 30 g.

[0144] 7.3 Data Analysis The antitumor growth curves of the test substance, mouse survival time, and time to ascites formation were plotted with the X-axis representing treatment time (days) and the Y-axis representing the corresponding tumor volume (mean), survival rate, and rate of no ascites formation. Intergroup differences in tumor volume were compared using one-way ANOVA, and if there was a significant difference in the F-score (a ratio of treatment variance to the error variance), intergroup comparisons were performed using the Games-Howell test. Statistical analysis was performed on the data using SPSS (Statistical Product and Service Solutions) software (version 18.0, IBM, Armonk, NY, US). The data were plotted using Prism version 9 software (GraphPad Software Inc., San Diego, CA), and survival time and time to ascites formation were analyzed using the log-rank test. A synergistic effect score of <1 for concomitant administration indicated an antagonistic effect, =1 indicated an additive effect, and >1 indicated a synergistic effect.

[0145] 7.4 Results This experiment evaluated the combined therapeutic effect of the compound represented by formula (I-1) and PLD in the ID8 model. The compound represented by formula (I-1) was administered at a dose of 100 mg / kg for 21 days in the pre- and qd phases, while PLD was administered at a dose of 3 mg / kg for 3 weeks in the iv and qw phases.

[0146] On day 22, the tumor burden of mice was measured, and as shown in Figure 9, the T / C ratios for the monotherapy group of the compound represented by formula (I-1), the monotherapy group of PLD, and the combination therapy group were 37.49%, 16.93%, and 6.65%, respectively. As shown in Table 6, the monotherapy group of the compound represented by formula (I-1) had a T / C value of 97.13% on day 36. The monotherapy group of PLD had a T / C value of 33.39% on day 36. The combination therapy group of the compound represented by formula (I-1) and PLD had a T / C value of 15.13% on day 36, and the synergistic effect coefficient of the two combined administrations was high, reaching 2.14. This demonstrated that the combination therapy of the two drugs had a strong inhibitory effect on tumor growth. Ascites, which occurs when the model progresses in the later stages, affects the accuracy of imaging, so drug efficacy was evaluated by detecting the abdominal circumference, progression of ascites, and survival time of mice instead of in vivo imaging. As shown in Figure 10, compared to the solvent control group, the survival time of mice was extended in both the group treated with the compound represented by formula (I-1) and the group treated with PLD. Furthermore, the survival time of mice in the dual-drug combination group was significantly extended compared to both the solvent control group and the monotherapy group, demonstrating that the dual-drug combination has a synergistic effect in extending the survival time of mice. As shown in Figures 11 and 12, the progression of ovarian cancer in mice was characterized by the detection of ascites production and the measurement of the abdominal circumference of the mice, and it was found that the dual-drug combination can significantly extend the progression-free survival time of mice compared to monotherapy. As shown in Figure 13, during the treatment process, no serious weight loss was observed in the animals of each group, and the animals were in good condition.

[0147] In summary, in the ID8 model, the antitumor effect of the compound shown in formula (I-1) in combination with PLD is superior to that of the compound shown in formula (I-1) or PLD alone.

[0148] [Table 6]

[0149] Example 4 Title: Treatment of platinum-resistant recurrent ovarian cancer or advanced solid tumors in patients with APG-2449 monotherapy or in combination with PLD.

[0150] Study Objectives: This study is an open, multicenter, dose-finding Phase I clinical trial intended to evaluate the safety of APG-2449 monotherapy in patients with advanced solid tumors, and the safety, resistance, and efficacy of APG-2449 in combination with PLD in the treatment of ovarian cancer.

[0151] Research design: This study includes the following two parts: Part A: Treatment of advanced solid tumors with APG-2449 monotherapy. APG-2449 is administered orally once daily (QD) at a dose of 1200 mg as monotherapy (RP2D) after meals, in a 28-day administration cycle, to determine its safety and pharmacokinetic characteristics.

[0152] Part B: Dosage considerations and expansion of the combination therapy of APG-2449 and PLD. The drug APG-2449 is administered orally once daily (QD) at a dose of 1200 mg per dose, with one administration cycle consisting of 28 days. The drug PLD is administered at 40 mg / m². 2 It is administered by intravenous drip infusion, with each administration cycle lasting 28 days, and an injection is given on the first day of each cycle.

[0153] Using a standard "3+3" design, the subsequent treatment group will either increase or decrease the APG-2449 dose to 1500 mg or 900 mg. Additionally, two doses will be selected for expansion to evaluate the efficacy of the combination therapy.

[0154] Observation indicators: 1. Treatment-related adverse events in NCI-CTCAE version 5.0.

[0155] Based on CTCAE v5.0, the number and frequency of adverse events for the study drug were evaluated. This included the number of patients who experienced adverse events, and the number of patients who experienced abnormal vital signs, abnormal health examinations, abnormal laboratory findings, and abnormal 12-lead electrocardiogram findings in APG-2449 monotherapy and in combination therapy with PLD.

[0156] 2. Dose-limiting toxicity (DLT).

[0157] DLT was defined by the incidence of drug-related grade 3 to grade 5 adverse events occurring within 4 weeks prior to the study treatment. These were assessed according to the NCI-CTCAE 5.0 version.

[0158] Although specific embodiments of the present invention have been described above, those skilled in the art should understand that these are merely illustrative descriptions and that various changes or modifications can be made to these embodiments without departing from the principles and substance of the present invention. Accordingly, the scope of protection of the present invention is limited by the appended claims.

Claims

1. It is a combination of medicines, Substance X is a compound represented by formula (I), a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof. The substance Y comprises doxorubicin, a pharmaceutically acceptable salt thereof, its solvate, and a solvate of its pharmaceutically acceptable salt. 【Chemistry 1】 Eventually, R １ａ and R ２ａ These are hydrogen and C, independently. １－４ Alkyl alkyl group or C ３－６ It is a cycloalkyl group, R ３ teeth 【Chemistry 2】 That is, Combinations of pharmaceuticals.

2. (1) The aforementioned combination of pharmaceuticals is a combination of pharmaceuticals for treating and / or preventing ovarian cancer, (2) The substance X is in a therapeutically effective amount, (3) The substance Y is in a therapeutically effective amount, (4) The substance X is a compound represented by formula (I) or a pharmaceutically acceptable salt thereof, for example, a compound represented by formula (I), and preferably the compound represented by formula (I) is a compound represented by formula (I-1), 【Transformation 3】 (5) The substance Y is doxorubicin or a pharmaceutically acceptable salt thereof, for example, a pharmaceutically acceptable salt of doxorubicin, preferably doxorubicin hydrochloride or PLD. (6) The mass ratio of substance X to substance Y within one administration cycle of 28 days is (200-1000):1, preferably (300-900):1, more preferably (300-800):

1. It is characterized by satisfying one or more of the following conditions: The pharmaceutical combination described in claim 1.

3. The pharmaceutical combination according to claim 1, characterized in that the substance X is a compound represented by formula (I-1), the substance Y is doxorubicin hydrochloride or PLD, and the mass ratio of substance X to substance Y within a 28-day administration cycle is (200-1000):1, preferably (300-900):1, and more preferably (300-800):

1.

4. (1) The dosage of substance X is 50 mg / kg to 200 mg / kg, for example 80 mg / kg to 120 mg / kg, preferably 100 mg / kg, or the dosage is 600 mg to 2000 mg, preferably 900 mg to 1500 mg, more preferably 1200 mg, and the dosage may be in the form of a single dose or multiple doses. (2) The administration frequency of substance X is once a day, (3) The method of administering the substance X is intragastric administration or oral administration, for example intragastric administration. (4) The dosage of substance Y is 1 mg / kg to 15 mg / kg, for example 2 mg / kg to 10 mg / kg, preferably 3 mg / kg, or the dosage is 40 mg / m ２ The dose is administered by intravenous drip infusion, for example, 40 mg to 80 mg, or for example, 50 mg to 70 mg, and this dose may be in the form of a single dose or multiple doses. (5) The administration frequency of substance Y is once a week or once every 28 days. (6) The method of administering the substance Y is intravenous injection, for example, intravenous bolus injection. (7) Substances X and Y may be used simultaneously, separately, or sequentially. It is characterized by satisfying one or more of the following conditions: The pharmaceutical combination described in claim 1.

5. The pharmaceutical combination according to at least one of claims 1 to 4, characterized in that the administration frequency of substance X is once a day, the administration frequency of substance Y is once a week with a three-week cycle as one period, or the administration frequency of substance Y is once every 28 days.

6. The pharmaceutical combination according to claim 5, characterized in that the dosage of substance X is 100 mg / kg or 600 mg to 2000 mg, administered once a day, the dosage of substance Y is 3 mg / kg, administered once a week with a 3-week cycle, or the dosage of Y is 40 mg to 80 mg, administered once every 28 days.

7. The pharmaceutical combination according to claim 5, characterized in that the dose of substance X is 100 mg / kg or 600 mg to 2000 mg, the frequency of administration is once a day, and the method of administration is intragastric or oral, the dose of substance Y is 3 mg / kg, the frequency of administration is once a week, the method of administration is intravenous bolus injection, with a cycle of three weeks, or the dose of Y is 40 mg to 80 mg, administered once every 28 days, and the method of administration is intravenous bolus injection.

8. The pharmaceutical combination according to claim 5, characterized in that the substance X is a compound represented by formula (I-1), the dose of substance X is 100 mg / kg or 600 mg to 2000 mg, the frequency of administration is once a day, and the method of administration is intragastric or oral, and the substance Y is doxorubicin hydrochloride or PLD, the dose of substance Y is 3 mg / kg, the frequency of administration is once a week, the method of administration is intravenous bolus injection, with a cycle of three weeks, or the dose of Y is 40 mg to 80 mg, administered once every 28 days, and the method of administration is intravenous bolus injection.

9. Pharmaceutical composition A, A first pharmaceutical composition comprising substance X and a medicinal auxiliary material, wherein substance X is a compound represented by formula (I) as described in claim 1, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof. Furthermore, a second pharmaceutical composition comprising substance Y and a medicinal auxiliary material, wherein substance Y is doxorubicin, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof, Pharmaceutical composition A.

10. (1) The pharmaceutical composition A is a pharmaceutical composition A for treating and / or preventing ovarian cancer, (2) In the first pharmaceutical composition, the substance X is in a therapeutically effective amount, (3) In the second pharmaceutical composition described above, the substance Y is in a therapeutically effective amount, (4) In the first pharmaceutical composition, the substance X is a compound represented by formula (I) or a pharmaceutically acceptable salt thereof, for example, a compound represented by formula (I), and preferably the compound represented by formula (I) is a compound represented by formula (I-1), 【Chemistry 4】 (5) In the second pharmaceutical composition, the substance Y is doxorubicin or a pharmaceutically acceptable salt thereof, for example, a pharmaceutically acceptable salt of doxorubicin, preferably doxorubicin hydrochloride. It is characterized by satisfying one or more of the following conditions: Pharmaceutical composition A as described in claim 9.

11. The pharmaceutical composition A according to claim 9, characterized in that the second pharmaceutical composition is a substance Y liposome.

12. (1) In the first pharmaceutical composition, the dose of substance X is 50 mg / kg to 200 mg / kg, for example 80 mg / kg to 120 mg / kg, preferably 100 mg / kg, or the dose is 600 mg to 2000 mg, preferably 900 mg to 1500 mg, more preferably 1200 mg, and the dose may be in the form of a single dose or a multiple dose. (2) The frequency of administration of the first pharmaceutical composition is once a day, (3) The method of administering the first pharmaceutical composition is intragastric administration or oral administration. (4) The second pharmaceutical composition is substance Y liposome, and the dosage of substance Y liposome is 1 mg / kg to 15 mg / kg, for example 2 mg / kg to 10 mg / kg, preferably 3 mg / kg, or the dosage is 40 mg / m ２ The dose is administered by intravenous drip infusion, for example, 40 mg to 80 mg, or for example, 50 mg to 70 mg, and this dose may be in the form of a single dose or multiple doses. (5) The frequency of administration of the second pharmaceutical composition is once a week or once every 28 days. (6) The method of administering the second pharmaceutical composition is intravenous injection, for example, intravenous bolus injection. (7) The first pharmaceutical composition and the second pharmaceutical composition are administered simultaneously, separately, or sequentially. (8) The second pharmaceutical composition is polyethylene glycol liposome doxorubicin. It is characterized by satisfying one or more of the following conditions: Pharmaceutical composition A as described in claim 9.

13. The pharmaceutical composition A according to claim 9, characterized in that the second pharmaceutical composition is a substance Y liposome, and the mass ratio of substance X to substance Y liposome in the first pharmaceutical composition within a 28-day administration cycle is (200-1000):1, preferably (300-900):1, and more preferably (300-800):

1.

14. A first pharmaceutical composition comprising substance X and a medicinal auxiliary material, wherein substance X is a compound represented by formula (I-1), and, A second pharmaceutical composition comprising polyethylene glycol liposome doxorubicin, The mass ratio of the compound represented by formula (I-1) to the polyethylene glycol liposomal doxorubicin during one administration cycle of 28 days is (200-1000):1, preferably (300-900):1, and more preferably (300-800):1, characterized in that The pharmaceutical combination A described in claim 9.

15. The pharmaceutical composition A according to at least one of claims 9 to 14, characterized in that the first pharmaceutical composition is administered once a day, and the second pharmaceutical composition is administered once a week with a three-week cycle, or once every 28 days.

16. The pharmaceutical composition A according to claim 15, characterized in that, in the first pharmaceutical composition, the dose of substance X is 100 mg / kg or 600 mg to 2000 mg, and the frequency of administration of the first pharmaceutical composition is once a day, and the second pharmaceutical composition is substance Y liposome, the dose of substance Y liposome is 3 mg / kg, and the frequency of administration of the second pharmaceutical composition is once a week, with a cycle of three weeks, or the dose of substance Y liposome is 40 mg to 80 mg, once every 28 days.

17. The pharmaceutical composition A according to claim 15, characterized in that, in the first pharmaceutical composition, the dose of substance X is 100 mg / kg or 600 mg to 2000 mg, the method of administration of the first pharmaceutical composition is intragastric or oral administration, and the frequency of administration is once a day, the second pharmaceutical composition is substance Y liposome, the dose of substance Y liposome is 3 mg / kg or 40 mg to 80 mg, the method of administration of the second pharmaceutical composition is intravenous bolus injection, and the frequency of administration is once a week with a cycle of three weeks, or once every 28 days.

18. The first pharmaceutical composition is characterized in that, in the first pharmaceutical composition, substance X is a compound represented by formula (I-1), the dose of substance X is 100 mg / kg or 600 mg to 2000 mg, the method of administering the first pharmaceutical composition is intragastric or oral administration, and the frequency of administration is once a day, and the second pharmaceutical composition is polyethylene glycol liposomal doxorubicin, the dose of polyethylene glycol liposomal doxorubicin is 3 mg / kg or 40 mg to 80 mg, the method of administering polyethylene glycol liposomal doxorubicin is intravenous bolus injection, and the frequency of administration is once a week with a cycle of three weeks, or once every 28 days, as described in claim 15.

19. It is a combination kit, A first container comprising the first pharmaceutical composition according to at least one of claims 9 to 18, A second container comprising the second pharmaceutical composition according to at least one of claims 9 to 18, Combination kit.

20. The use of a pharmaceutical combination according to at least one of claims 1 to 8 or pharmaceutical composition A according to at least one of claims 9 to 18 in the manufacture of a drug for preventing and / or treating ovarian cancer.

21. A method for preventing and / or treating ovarian cancer, comprising administering to a patient in need of such treatment a pharmaceutical combination according to at least one of claims 1 to 8 or a pharmaceutical composition A according to at least one of claims 9 to 18.

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