Use of a pharmaceutical composition in the treatment of cancer

Abstract

The application discloses application of a pharmaceutical composition in treating cancer, wherein the pharmaceutical composition comprises a BCL-XL inhibitor and a HER2 targeted drug or comprises a BCL-XL inhibitor and a STAT3 inhibitor. The application provides medical use of the pharmaceutical composition according to the application for treating or preventing cancer. The application also provides a method for treating or preventing cancer, and provides a brand-new treatment scheme for the technical field of cancer treatment, and has important scientific significance and clinical application value.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to the use of a pharmaceutical composition in treating cancer, and more specifically, to the use of a BCL-XL inhibitor and a HER2-targeted drug in combination in the preparation of a drug for treating cancer. Background Art

[0002] Cancer is one of the most devastating diseases to human health, claiming a significant number of lives each year. Currently, cancer is typically treated with surgery, radiotherapy, chemotherapy, and medication. However, surgery alone is ineffective, resulting in a poor prognosis for some patients with advanced disease. Consequently, there is an unmet need for more effective cancer treatment strategies.

[0003] BCL-XL inhibitors are a class of drugs that inhibit the function of the BCL-XL (B-cell lymphoma / leukemia-2 long homolog X) protein. By inhibiting BCL-XL function, they promote cell apoptosis and prevent the growth and spread of cancer cells. Recent studies have shown that BCL-XL inhibitors have excellent anti-cancer effects, including positive effects on non-Hodgkin's lymphoma and diabetic macular edema.

[0004] HER2-targeted drugs are therapeutic agents for HER2 (human epidermal growth factor receptor 2)-positive cancers. They specifically bind to the HER2 receptor, inhibiting HER2-mediated signaling, including cell proliferation and differentiation, thereby suppressing tumor growth. In clinical treatment, combining these drugs with chemotherapy or radiotherapy can enhance therapeutic efficacy. However, there are currently no reports on the combined treatment of cancer with BCL-XL inhibitors and HER2-targeted drugs. Summary of the Invention

[0005] In view of this, in order to solve the above-mentioned technical problems currently faced in this field, the purpose of the present invention is to provide a combination pharmaceutical composition for treating cancer, thereby providing an effective drug combination strategy for the treatment of cancer.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] A first aspect of the present invention provides a pharmaceutical composition for preventing and / or treating cancer.

[0008] Furthermore, the pharmaceutical composition comprises a BCL-XL inhibitor and a HER2 targeting drug.

[0009] Furthermore, the BCL-XL inhibitor is selected from any one of A1331852 or a pharmaceutically acceptable salt thereof, and ABT263 or a pharmaceutically acceptable salt thereof.

[0010] Furthermore, the HER2 targeted drug is selected from any one of Trastuzumab or a pharmaceutically acceptable salt thereof, Lapatinib or a pharmaceutically acceptable salt thereof, and Neratinib or a pharmaceutically acceptable salt thereof.

[0011] Furthermore, the cancer includes gastroesophageal adenocarcinoma, breast cancer, ovarian cancer, bladder cancer, lung cancer, colon cancer, rectal cancer, prostate cancer, blood cancer, lymphoma, cervical cancer, osteosarcoma, glioblastoma, melanoma, pancreatic cancer, liver cancer, kidney cancer, gallbladder cancer, bile duct cancer, esophageal cancer or neuroblastoma.

[0012] Furthermore, the cancer is selected from gastroesophageal adenocarcinoma, breast cancer, ovarian cancer, and bladder cancer.

[0013] Furthermore, when the cancer is gastroesophageal adenocarcinoma, the concentration ratio of A1331852 to Lapatinib is (12.5 nM~200 nM: 62.5 nM~1000 nM); the concentration ratio of A1331852 to Neratinib is (12.5 nM~200 nM: 31.5 nM~500 nM); the concentration ratio of A1331852 to Trastuzumab is (12.5 nM~200 nM: 12.5 ng / mL~200 ng / mL);

[0014] Furthermore, when the cancer is breast cancer, the concentration ratio of A1331852 to Lapatinib is (12.5 nM~200 nM: 62.5 nM~1000 nM); the concentration ratio of A1331852 to Neratinib is (12.5 nM~200 nM: 12.5 nM~200 nM); the concentration ratio of A1331852 to Trastuzumab is (12.5 nM~200 nM: 12.5 ng / mL~200 ng / mL);

[0015] Furthermore, when the cancer is ovarian cancer, the concentration ratio of A1331852 to Lapatinib is (0.625 μM~10 μM:0.625 μM~10 μM); the concentration ratio of A1331852 to Neratinib is (0.625 μM~10 μM:62.5 nM~1000 nM); the concentration ratio of A1331852 to Trastuzumab is (0.625 μM~10 μM:12.5 ng / mL~200 ng / mL);

[0016] Furthermore, when the cancer is bladder cancer, the concentration ratio of A1331852 to Lapatinib is (12.5nM~200nM:62.5nM~1000nM); the concentration ratio of A1331852 to Neratinib is (12.5nM~200nM:12.5nM~200nM); and the concentration ratio of A1331852 to Trastuzumab is (12.5nM~200nM:12.5ng / mL~200ng / mL).

[0017] Furthermore, when the cancer is gastroesophageal adenocarcinoma, the concentration ratio of ABT263 to Lapatinib is (0.04 μM~10 μM: 62.5 nM~1000 nM); the concentration ratio of ABT263 to Neratinib is (0.04 μM~10 μM: 31.5 nM~500 nM); the concentration ratio of ABT263 to Trastuzumab is (0.04 μM~10 μM: 12.5 ng / mL~200 ng / mL);

[0018] Furthermore, when the cancer is breast cancer, the concentration ratio of ABT263 to Lapatinib is (0.04 μM~10 μM:62.5 nM~1000 nM); the concentration ratio of ABT263 to Neratinib is 0.04 μM~10 μM:31.5 nM~500 nM); the concentration ratio of ABT263 to Trastuzumab is (0.04 μM~10 μM:12.5 ng / mL~200 ng / mL);

[0019] Furthermore, when the cancer is ovarian cancer, the concentration ratio of ABT263 to Lapatinib is (0.04 μM~10 μM: 0.625 μM~10 μM); the concentration ratio of ABT263 to Neratinib is (0.04 μM~10 μM: 12.5 nM~200 nM); the concentration ratio of ABT263 to Trastuzumab is (0.04 μM~10 μM: 12.5 ng / mL~200 ng / mL);

[0020] Furthermore, when the cancer is bladder cancer, the concentration ratio of ABT263 to Lapatinib is (0.04 μM~10 μM: 62.5 nM~1000 nM); the concentration ratio of ABT263 to Neratinib is (0.04 μM~10 μM: 12.5 nM~200 nM); and the concentration ratio of ABT263 to Trastuzumab is (0.04 μM~10 μM: 12.5 ng / mL~200 ng / mL).

[0021] In a specific embodiment of the present invention, the inventors of the present invention discovered for the first time that the combination of a BCL-XL inhibitor and a HER2-targeted drug has a synergistic effect in the treatment of various cancers, including gastroesophageal adenocarcinoma, breast cancer, ovarian cancer, and bladder cancer. The present invention has demonstrated through experiments that the combination of the two can significantly inhibit the growth of cancer cells, exhibiting a significant synergistic killing effect.

[0022] In some embodiments, the BCL-XL inhibitor comprises any agent capable of inhibiting BCL-XL. For example, the BCL-XL inhibitor includes but is not limited to A1331852, ABT263, BH3I-, A1155463, and WEHI-539 hydrochloride. In a specific embodiment of the present invention, the BCL-XL inhibitor is selected from A1331852 and ABT263.

[0023] In some embodiments, the HER2-targeted drug includes any agent capable of inhibiting HER2. Exemplarily, the HER2-targeted drug includes but is not limited to: Lapatinib, Neratinib, Trastuzumab, Pertuzumab, T-DM1, DS-B201, Neratinib, and Pyrotinib. In a specific embodiment of the present invention, the HER2-targeted drug is selected from Lapatinib, Neratinib, and Trastuzumab.

[0024] In some embodiments, the pharmaceutically acceptable salts include acidic salts formed with inorganic and / or organic acids and basic salts formed with inorganic and / or organic bases. Furthermore, when the compound contains a basic moiety (such as, but not limited to, pyridine or imidazole) and an acidic moiety (such as, but not limited to, a carboxylic acid), zwitterions may form, and such zwitterions are included in the pharmaceutically acceptable salts described herein. Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, but other salts are also useful. Pharmaceutically acceptable salts of the compounds can be formed, for example, by reacting the compound with an amount of an acid or base in a medium, such as a medium in which the salt precipitates or an aqueous medium (followed by lyophilization).

[0025] Specific pharmaceutically acceptable salts include those salts that are, within the scope of sound medical judgment, suitable for contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic reaction, etc., and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts (pharmaceutically acceptable salts) are well known in the art. Pharmaceutically acceptable salts of the compounds of the present invention include salts derived from suitable inorganic and organic acids and inorganic and organic bases.

[0026] Examples of pharmaceutically acceptable nontoxic acid addition salts are salts formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid, or salts formed with organic acids, such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid. Salts formed using conventional methods in the art, such as ion exchange methods, are also included. Other pharmaceutically acceptable salts include: adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, gluconate, glycerophosphate, hemisulfate, heptanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate. Pharmaceutically acceptable salts derived from appropriate bases include alkali metal, alkaline earth metal, and ammonium salts. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium salts, and the like.

[0027] In the present invention, the term "preventing and / or treating" refers to delaying the development of a disease, preventing the development of a disease, and / or reducing the severity of symptoms that will develop or are expected to develop. Thus, these terms include ameliorating existing disease symptoms, preventing additional symptoms, ameliorating or preventing potential metabolic causes of symptoms, inhibiting the disorder or disease, e.g., arresting the development of the disorder or disease, alleviating the disorder or disease, causing the disorder or disease to regress, alleviating symptoms caused by the disease or disorder, or halting symptoms of the disease or disorder.

[0028] Furthermore, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier and / or excipient.

[0029] In some embodiments, the pharmaceutically acceptable carriers and / or excipients include, but are not limited to, diluents, binders, surfactants, wetting agents, adsorption carriers, lubricants, fillers, and disintegrants.

[0030] In some embodiments, the diluent includes, but is not limited to, lactose, sodium chloride, glucose, urea, starch, water, and the like.

[0031] In some embodiments, the binder includes but is not limited to starch, pregelatinized starch, dextrin, maltodextrin, sucrose, gum arabic, gelatin, methyl cellulose, carboxymethyl cellulose, ethyl cellulose, polyvinyl alcohol, polyethylene glycol, polyvinyl pyrrolidone, alginic acid and alginates, xanthan gum, hydroxypropyl cellulose and hydroxypropyl methylcellulose, etc.

[0032] In some embodiments, the surfactant includes, but is not limited to, polyoxyethylene sorbitan fatty acid ester, sodium lauryl sulfate, stearic acid monoglyceride, cetyl alcohol, and the like.

[0033] In some embodiments, the humectant includes but is not limited to glycerol, starch, and the like.

[0034] In some embodiments, the adsorption carrier includes, but is not limited to, starch, lactose, bentonite, silica gel, kaolin, and bentonite.

[0035] In some embodiments, the lubricant includes but is not limited to: zinc stearate, glyceryl monostearate, polyethylene glycol, talc, calcium and magnesium stearate, polyethylene glycol, boric acid powder, hydrogenated vegetable oil, sodium stearyl fumarate, polyoxyethylene monostearate, monolauric sucrose ester, sodium lauryl sulfate, magnesium lauryl sulfate, magnesium dodecyl sulfate, etc.

[0036] In some embodiments, the filler includes but is not limited to: mannitol (granular or powdered), xylitol, sorbitol, maltose, erythrose, microcrystalline cellulose, polymeric sugars, coupling sugars, glucose, lactose, sucrose, dextrin, starch, sodium alginate, laminarin powder, agar powder, calcium carbonate, and sodium bicarbonate.

[0037] In some embodiments, the disintegrant includes but is not limited to: cross-linked vinyl pyrrolidone, sodium carboxymethyl starch, low-substituted hydroxypropyl methyl, cross-linked sodium carboxymethyl cellulose, soybean polysaccharide, etc.

[0038] In some embodiments, the pharmaceutically acceptable carrier and / or excipient is used as needed to help stabilize the formulation, improve its activity or bioavailability, or produce an acceptable taste or smell when taken orally. The pharmaceutical composition thus formulated can be administered by any appropriate route known to those skilled in the art. When the pharmaceutical composition is used, a safe and effective amount of the pharmaceutical composition of the present invention is administered to a human.

[0039] In some embodiments, the appropriate dosage of the pharmaceutical composition of the present invention can be prescribed in a variety of ways depending on factors such as the formulation method, administration method, patient's age, weight, sex, morbidity, diet, administration time, administration route, excretion rate and reaction sensitivity. A skilled physician can usually easily determine the prescription and the desired effective dosage for treatment.

[0040] Furthermore, the pharmaceutical composition of the present invention may further include additives such as stabilizers, bactericides, buffers, isotonic agents, chelating agents, pH control agents and surfactants.

[0041] Furthermore, the pharmaceutical compositions of the present invention may be used in combination with other drugs for treating cancer. The other therapeutic compounds may be administered concurrently with the principal active ingredient, or even in the same composition. The other therapeutic compounds may also be administered separately, either as separate compositions or in dosage forms different from those of the principal active ingredient. A portion of the principal ingredient may be administered concurrently with the other therapeutic compound, while the remaining portion may be administered alone. During treatment, the dosage of the pharmaceutical compositions of the present invention may be adjusted based on the severity of symptoms, the frequency of relapses, and the physiological response to the treatment regimen.

[0042] Furthermore, the dosage form of the pharmaceutical composition includes a non-gastrointestinal dosage form and / or a gastrointestinal dosage form.

[0043] Furthermore, the non-gastrointestinal administration dosage form includes an injection dosage form, a cavity administration dosage form, a mucosal administration dosage form and / or a skin administration dosage form.

[0044] Furthermore, the dosage forms for administration via the gastrointestinal tract include tablets, granules, capsules, solutions, powders, sustained-release preparations, emulsions, suspensions, syrups and / or drops.

[0045] In some embodiments, the dosage form of the pharmaceutical composition of the present invention is a dosage form that is prepared by conventional methods and is convenient for administration, including but not limited to: parenteral dosage forms, gastrointestinal dosage forms, and specific examples include but are not limited to: aqueous solution injection, powder injection, pills, powders, tablets, patches, suppositories, emulsions, creams, gels, granules, capsules, aerosols, sprays, powder sprays, sustained-release agents and controlled-release agents, etc.

[0046] In some embodiments, the injection dosage forms include, but are not limited to, intravenous injections, intramuscular injections, subcutaneous injections, intradermal injections, intracavitary injections, and other injections.

[0047] In some embodiments, the cavity administration dosage form includes but is not limited to: suppositories, aerosols, effervescent tablets, drops, pills, etc., which are used in the rectum, vagina, urethra, nasal cavity, ear canal, etc.

[0048] In some embodiments, the mucosal administration dosage form includes, but is not limited to, eye drops, nasal drops, eye ointments, gargles, sublingual tablets, adhesive tablets, patches, and the like.

[0049] In some embodiments, the skin administration dosage form includes, but is not limited to, topical solutions, lotions, liniments, ointments, plasters, pastes, patches, and the like.

[0050] In some embodiments, the pharmaceutical composition includes pharmaceutical excipients. These can be conventionally used in various pharmaceutical preparations, including but not limited to isotonicity agents, buffers, flavoring agents, excipients, fillers, binders, disintegrants, and lubricants. They can also be selected for compatibility with the substance, including but not limited to emulsifiers, solubilizers, antibacterial agents, analgesics, and antioxidants. Such excipients can effectively improve the stability and solubility of the active ingredients contained in the composition or modify the release and absorption rates of the active ingredients, thereby improving the metabolism of the active ingredients in the body and enhancing the administration effect of the composition. Furthermore, excipients can be used to achieve specific administration purposes or modes, such as sustained-release, controlled-release, and pulse administration, and include but are not limited to gelatin, albumin, chitosan, and polyether and polyester polymers (e.g., polyethylene glycol, polyurethane, polycarbonate, and their copolymers). These excipients can enhance administration by improving therapeutic efficacy, increasing bioavailability, reducing toxic side effects, and improving patient compliance.

[0051] In some embodiments, the pharmaceutical composition or pharmaceutical preparation may also contain other drugs or agents that can be used for treatment and / or prevention, or adjuvant treatment and / or prevention. There are no special restrictions on the other drugs or agents. As long as they can produce the expected treatment and / or prevention, or adjuvant treatment and / or prevention effect on cancer, they are within the scope of protection of the present invention.

[0052] In some embodiments, the other drugs or agents that can be used for treatment and / or prevention, or adjuvant treatment and / or prevention include but are not limited to: chemotherapy drugs, specifically, the chemotherapy drugs include but are not limited to: taxanes (paclitaxel, docetaxel, cabazitaxel), antibiotics (doxorubicin, epirubicin, daunorubicin, pirarubicin, etoposide, irinotecan, mitoxantrone), antimetabolites (cisplatin, carboplatin, lobaplatin, nedaplatin, oxaliplatin, gemcitabine, 5-fluorouracil), alkylating agents (cyclophosphamide, ifosfamide).

[0053] Furthermore, the pharmaceutical composition is a single compound preparation or a combination of two separate single preparations.

[0054] Furthermore, the compound preparation is a compound preparation comprising a BCL-XL inhibitor and a HER2 targeted drug.

[0055] Furthermore, the combination of single-ingredient preparations is a combination of a single-ingredient preparation comprising a BCL-XL inhibitor and a single-ingredient preparation comprising a HER2-targeted drug.

[0056] Furthermore, the BCL-XL inhibitor is selected from any one of A1331852 or a pharmaceutically acceptable salt thereof, and ABT263 or a pharmaceutically acceptable salt thereof.

[0057] Furthermore, the HER2 targeted drug is selected from any one of Trastuzumab or a pharmaceutically acceptable salt thereof, Lapatinib or a pharmaceutically acceptable salt thereof, and Neratinib or a pharmaceutically acceptable salt thereof.

[0058] In some embodiments, the compound preparation refers to a preparation made of two or more active pharmaceutical ingredients. For example, when the pharmaceutical composition of the present invention is a compound preparation, it may mean that it contains both a BCL-XL inhibitor and a HER2 targeting drug.

[0059] In some embodiments, the single-ingredient preparation refers to a preparation made with a single active pharmaceutical ingredient. For example, when the pharmaceutical composition of the present invention is a combination of single-ingredient preparations, it may represent a combination of a single-ingredient preparation containing a BCL-XL inhibitor and a single-ingredient preparation containing a HER2-targeted drug.

[0060] In some embodiments, the HER2-targeted drug and BCL-XL inhibitor in the pharmaceutical composition can be administered simultaneously, separately, or sequentially. Simultaneously refers to the simultaneous administration of the two drugs. If not administered simultaneously, they are administered sequentially within a timeframe such that both drugs can act therapeutically within the same timeframe. Thus, sequential administration allows for administration of one drug within 5 minutes, 10 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, or several hours of one drug, provided that the circulating half-life of the first administered drug allows both drugs to be present in therapeutically effective amounts simultaneously. The time delay between administration of the components will vary depending on the precise nature of the components, their interactions, and their respective half-lives. Unlike simultaneous or sequential administration, separate administration refers to administration of one drug with a significant interval between the administration of the other, i.e., the first administered drug may no longer be present in the bloodstream in therapeutically effective amounts when the second drug is administered.

[0061] A second aspect of the present invention provides a pharmaceutical composition for preventing and / or treating gastroesophageal adenocarcinoma.

[0062] Furthermore, the pharmaceutical composition comprises a BCL-XL inhibitor and a STAT3 inhibitor.

[0063] Preferably, the BCL-XL inhibitor is selected from A1331852 or a pharmaceutically acceptable salt thereof; and the STAT3 inhibitor is selected from Napabucasin or a pharmaceutically acceptable salt thereof.

[0064] Preferably, when phosphorylated STAT3 is highly expressed in the gastroesophageal adenocarcinoma, the concentration ratio of A1331852 to Napabucasin is (0.002 μM-10 μM: 80 nM-300 nM).

[0065] Preferably, when phosphorylated STAT3 is lowly expressed in the gastroesophageal adenocarcinoma, the concentration ratio of A1331852 to Napabucasin is (25 nM-400 nM: 50 nM-1000 nM).

[0066] The third aspect of the present invention provides an in vitro non-therapeutic method for inhibiting cancer cells or promoting apoptosis of cancer cells.

[0067] Furthermore, the method comprises: treating cancer cells with the pharmaceutical composition described in the first aspect of the present invention or the pharmaceutical composition described in the second aspect of the present invention.

[0068] Furthermore, the cancer cells include gastroesophageal adenocarcinoma cells, adenocarcinoma cells, ovarian cancer cells, bladder cancer cells, lung cancer cells, colon cancer cells, rectal cancer cells, prostate cancer cells, blood cancer cells, lymphoma cells, cervical cancer cells, osteosarcoma cells, glioblastoma cells, melanoma cells, pancreatic cancer cells, liver cancer cells, renal cancer cells, gallbladder cancer cells, bile duct cancer cells or neuroblastoma cells.

[0069] Furthermore, the cancer is selected from gastroesophageal adenocarcinoma cells, breast cancer cells, ovarian cancer cells, and bladder cancer cells.

[0070] In a specific embodiment of the present invention, the present invention has experimentally demonstrated that the pharmaceutical composition has a significant synergistic inhibitory effect on the growth and proliferation of gastroesophageal adenocarcinoma cells, breast cancer cells, ovarian cancer cells, and bladder cancer cells. Therefore, the pharmaceutical composition can be used as an inhibitor for inhibiting the growth and proliferation of gastroesophageal cancer cells, breast cancer cells, ovarian cancer cells, and bladder cancer cells for non-therapeutic purposes, and can be used in the field of scientific research, such as more in-depth research on the growth and metabolic mechanisms or behaviors of gastroesophageal adenocarcinoma cells, breast cancer cells, ovarian cancer cells, and bladder cancer cells, and screening for potential drugs that can be used to treat gastroesophageal adenocarcinoma cells, breast cancer cells, ovarian cancer cells, and bladder cancer cells.

[0071] The present invention also provides a method for treating and / or preventing cancer, comprising: administering a therapeutically and / or preventively effective amount of the pharmaceutical composition according to the first aspect of the present invention to a subject in need thereof.

[0072] Furthermore, the cancer includes gastroesophageal adenocarcinoma, breast cancer, ovarian cancer, bladder cancer, lung cancer, colon cancer, rectal cancer, prostate cancer, blood cancer, lymphoma, cervical cancer, osteosarcoma, glioblastoma, melanoma, pancreatic cancer, liver cancer, kidney cancer, gallbladder cancer, bile duct cancer or neuroblastoma.

[0073] Furthermore, the cancer is selected from gastroesophageal adenocarcinoma, breast cancer, ovarian cancer, and bladder cancer.

[0074] In the present invention, the subject includes mammals and non-mammals. Examples of mammals include, but are not limited to, any member of the class Mammalia: humans, non-human primates such as chimpanzees and other apes and monkeys; farm animals such as cattle, horses, sheep, goats, and pigs; domestic animals such as rabbits, dogs, and cats; and laboratory animals, including rodents such as rats, mice, and guinea pigs. Examples of non-mammals include, but are not limited to, birds and fish. In one embodiment of the present invention, the subject is a human. The term subject includes confirmed patients.

[0075] In the present invention, the effective amount refers to the amount of the compound effective to produce the desired preventive, allergic, or therapeutic effect. The amount of the pharmaceutical composition of the present invention required to achieve an effective amount will vary depending on factors such as the compound, the symptoms and their severity, and the age of the mammal being treated. However, the specific amount can be routinely determined by a person of ordinary skill in the art based on their knowledge in the art and the disclosure herein. Any dosage that can produce the aforementioned effects is within the scope of this invention.

[0076] In some embodiments, the pharmaceutical composition can be administered in any form, including but not limited to oral, topical, parenteral, intramuscular, intravenous, subcutaneous, transdermal (which may include a penetration enhancer), oral and / or suppository administration, as well as other routes of administration. In one embodiment of the invention, the active compound or combination of compounds as described above is provided in a solid dosage form known in the art. The most effective dosage form will depend on the bioavailability or pharmacokinetics of the selected particular agent and the severity of the patient's disease. Oral dosage forms are particularly preferred because of their ease of administration and expected favorable patient compliance.

[0077] In some embodiments, the pharmaceutical composition or pharmaceutical preparation can be administered to the subject by oral administration, injection, or topical administration. For example, the method can include administering the pharmaceutical composition or pharmaceutical preparation to the subject three times a day, once a day, once every two days, etc. In some embodiments, injection can include subcutaneous injection, intramuscular injection, intravenous injection, etc. In some embodiments, injection can include injecting the pharmaceutical composition directly into the lesion or in an area near the lesion. In some embodiments, a combination of different administration methods can be used.

[0078] The fourth aspect of the present invention provides any of the following applications.

[0079] Furthermore, the application includes:

[0080] (1) Use of a combination of a BCL-XL inhibitor and a HER2-targeted drug in the preparation of a drug for the prevention and / or treatment of cancer;

[0081] (2) The application of BCL-XL inhibitors and HER2-targeted drugs in combination to inhibit cancer cells or promote apoptosis of cancer cells for non-therapeutic purposes in vitro;

[0082] (3) The use of a combination of a BCL-XL inhibitor and a HER2-targeted drug in the preparation of a reagent for inhibiting cancer cells or cancer cell apoptosis in vitro;

[0083] (4) Use of BCL-XL inhibitors in the preparation of drugs for improving the therapeutic effect of HER2-targeted drugs in treating cancer;

[0084] (5) Application of HER2-targeted drugs in the preparation of drugs for improving the therapeutic effect of BCL-XL inhibitors in treating cancer;

[0085] (6) Use of a BCL-XL inhibitor and a STAT3 inhibitor in combination in the preparation of a drug for preventing and / or treating gastroesophageal adenocarcinoma;

[0086] (7) The combined use of BCL-XL inhibitors and STAT3 inhibitors in the in vitro non-therapeutic inhibition of gastroesophageal adenocarcinoma cells or promotion of apoptosis of gastroesophageal adenocarcinoma cells;

[0087] (8) Use of a BCL-XL inhibitor and a STAT3 inhibitor in combination for the preparation of a reagent for inhibiting apoptosis of gastroesophageal adenocarcinoma cells or gastroesophageal cancer cells in vitro;

[0088] (9) Use of BCL-XL inhibitors in the preparation of drugs for improving the therapeutic effect of STAT3 inhibitors in the treatment of gastroesophageal adenocarcinoma;

[0089] (10) Use of STAT3 inhibitors in the preparation of drugs for improving the therapeutic effect of BCL-XL inhibitors in the treatment of gastroesophageal adenocarcinoma.

[0090] Furthermore, the BCL-XL inhibitor is selected from any one of A1331852 or a pharmaceutically acceptable salt thereof, and ABT263 or a pharmaceutically acceptable salt thereof.

[0091] Furthermore, the HER2 targeting drug is selected from any one of Lapatinib or a pharmaceutically acceptable salt thereof, Trastuzumab or a pharmaceutically acceptable salt thereof, and Neratinib or a pharmaceutically acceptable salt thereof.

[0092] Furthermore, the STAT3 inhibitor is selected from Napabucasin or a pharmaceutically acceptable salt thereof.

[0093] Furthermore, the cancer includes gastroesophageal adenocarcinoma, breast cancer, ovarian cancer, bladder cancer, lung cancer, colon cancer, rectal cancer, prostate cancer, blood cancer, lymphoma, cervical cancer, osteosarcoma, glioblastoma, melanoma, pancreatic cancer, liver cancer, kidney cancer, gallbladder cancer, bile duct cancer, esophageal cancer or neuroblastoma.

[0094] Furthermore, the cancer is selected from gastroesophageal adenocarcinoma, breast cancer, ovarian cancer, and bladder cancer.

[0095] The present invention has the beneficial effects:

[0096] This invention, for the first time, discovers that the combination of a BCL-XL inhibitor and a HER2-targeted drug has a synergistic effect in the treatment of gastroesophageal adenocarcinoma, breast cancer, ovarian cancer, and bladder cancer. Furthermore, it finds that the combination of a BCL-XL inhibitor and a STAT3 inhibitor also has a synergistic effect in the treatment of gastroesophageal adenocarcinoma. This combined therapy significantly improves the therapeutic efficacy and achieves superior anti-tumor efficacy for these cancers. This invention provides a novel treatment option for these cancers, with significant scientific significance and clinical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0097] Figure 1 Show the effect of BCL-XL inhibitors combined with HER2-targeted drugs on the activity of gastroesophageal adenocarcinoma cells;

[0098] Figure 2 Show the effect of BCL-XL inhibitors combined with HER2-targeted drugs on breast cancer cell viability;

[0099] Figure 3 Show the effect of BCL-XL inhibitors combined with HER2-targeted drugs on the activity of ovarian cancer cells;

[0100] Figure 4 Show the effect of BCL-XL inhibitors combined with HER2-targeted drugs on the activity of bladder cancer cells;

[0101] Figure 5 Shows the effect of BCL-XL inhibitor combined with STAT3 inhibitor on the activity of gastroesophageal adenocarcinoma cells. DETAILED DESCRIPTION

[0102] Example 1 Effect of BCL-XL inhibitors combined with HER2 targeted drugs on cancer cell activity

[0103] 1. Experimental methods

[0104] (1) Cell plating

[0105] Drug sensitivity experiments were conducted using tumor cells in the logarithmic growth phase. The gastroesophageal adenocarcinoma cell line NCI-N87, the breast cancer cell line HCC1954, the ovarian cancer cell line SKOV3, and the bladder cancer cell line T24 were used. Cells were digested with trypsin-EDTA and counted. Using a multichannel pipette, 3,000 cells / 50 μL / well were evenly plated into a 96-well plate and incubated at 37°C with 5% CO2 until ready for use. To minimize edge effects, the outermost wells of the 96-well plate were not used for experiments and were filled with sterile PBS to minimize the effects of culture medium evaporation on cell growth and ensure experimental stability.

[0106] (2) Drug preparation

[0107] Drug concentration gradient design:

[0108] Drug A: HER2-targeted drugs. Trastuzumab, Lapatinib, and Neratinib were used in this project.

[0109] Drug B: BCL-XL inhibitors. A1331852 and ABT263 were used in this project.

[0110] Negative control: Trastuzumab solvent is PBS, and other drug solvents are DMSO. PBS or DMSO was used as negative control in this project.

[0111] Drug A and A1331852: Prepare the highest concentration using 4 times the actual application concentration, and then dilute the drug in a 1 / 2 gradient to prepare 5 concentrations.

[0112] ABT-263: Prepare the highest concentration using a dose 4 times the actual application concentration, and then dilute the drug in a 1 / 4 gradient to prepare 5 concentrations.

[0113] Negative control: Add PBS or DMSO in a volume corresponding to the highest drug concentration as a negative control.

[0114] Use a 1.8 mL 96-well plate to perform gradient dilutions of the above drugs.

[0115] (3) Drug treatment

[0116] Use a multichannel pipette to treat the cells plated in step (1) with the drug prepared in step (2).

[0117] Drug A concentration increasing direction: set to horizontal (row direction), 6 gradient concentrations, marked as C A (1-6);

[0118] Drug B concentration increasing direction: set to vertical (column direction), 6 titer concentrations, marked as C B (1-6).

[0119] The settings of the 96-well plate are shown in Table 1:

[0120] Table 1 Drug concentration settings

[0121]

[0122] The cell activity was detected after treatment with BCLXL inhibitors combined with Trastuzumab for 96 hours, and the cell activity was detected after treatment with BCLXL inhibitors combined with Lapatinib or Neratinib for 72 hours.

[0123] (4) Cell activity detection

[0124] In this project, the Promega CellTiter-Glo® Luminescent Cell Viability Assay Kit was used to detect cell viability. The specific procedures are as follows:

[0125] A. After removing the culture plate from the incubator, place it at room temperature for 10 minutes to allow it to equilibrate to room temperature.

[0126] B. Use a multichannel pipette to add 40 μL of CellTiter-Glo Reagent to each well and shake on a horizontal shaker at low speed (100–200 rpm) for 10 minutes.

[0127] C. Measure fluorescence intensity using an MD multi-function fluorescence microplate reader. Set the reading parameters to integration mode and the acquisition time of the fluorescence or luminescence signal to 1000 milliseconds (1 second) to ensure signal stability.

[0128] (5) Data analysis

[0129] 5.1 Calculate the percentage of cell viability:

[0130] Cell viability percentage = luminescence signal of drug-treated cells / luminescence signal of negative control cells × 100%

[0131] 5.2 Calculate the synergy score of the drug combination:

[0132] This project uses the Bliss independence model to evaluate the synergistic effect of the combination of drug A and drug B.

[0133] First, calculate the drug's inhibition rate on cell growth, inhibition rate = 1-cell activity percentage;

[0134] The predicted value of the cell growth inhibition rate of the combination of two drugs = (inhibition rate of drug A + inhibition rate of drug B - inhibition rate of drug A × inhibition rate of drug B) × 100%

[0135] Bliss synergy score = observed cell growth inhibition rate of the two drugs combined - predicted cell growth inhibition rate of the two drugs combined

[0136] If the Bliss synergy score is > 0, it indicates a synergistic effect; if the Bliss synergy score = 0, it indicates an additive effect; if the Bliss synergy score is < 0, it indicates an antagonistic effect.

[0137] 2. Experimental results

[0138] (1) Figure 1-5 Shows the percentage heatmap and Bliss synergy score heatmap of cell viability of BCL-XL inhibitors combined with HER2-targeted drugs in gastroesophageal adenocarcinoma, esophageal adenocarcinoma, breast cancer, ovarian cancer, and bladder cancer, respectively. Detailed descriptions are as follows:

[0139] Vertical axis: represents increasing concentrations of drug A (A1331852 or ABT263);

[0140] Horizontal axis: represents the increasing concentration of drug B (trastuzumab, lapatinib, or neratinib);

[0141] Cell activity percentage heat map white to red: indicates that the cell activity percentage decreases from 100 to 0.

[0142] Bliss synergy score heat map: blue indicates Bliss synergy score < 0, antagonistic effect, white indicates Bliss synergy score = 0, additive effect; red indicates Bliss synergy score > 0, synergistic effect.

[0143] Experimental results showed that BCL-XL inhibitors and HER2 targeted drugs had significant synergistic effects: the combination of A1331852 and HER2 targeted drugs showed strong synergistic effects in all different types of tumor cells; the combination of ABT263 and HER2 targeted drugs showed strong synergistic effects in gastroesophageal adenocarcinoma, esophageal adenocarcinoma, breast cancer and bladder cancer, and ABT263 and Lapatinib or Neratinib showed strong synergistic effects in ovarian cancer.

[0144] Example 2 Effects of BCL-XL Inhibitor Combined with STAT3 Inhibitor on the Activity of Gastroesophageal Adenocarcinoma Cells

[0145] 1. Experimental methods

[0146] (1) Cell plating

[0147] Drug sensitivity experiments were conducted using tumor cells in the logarithmic growth phase. Gastric adenocarcinoma cell lines were divided into two groups: the first group consisted of HGC-27, MKN-45, and AGS cell lines with high phosphorylated STAT3 expression, and the second group consisted of SNU-216, NCI-N87, and GCIY cell lines with low phosphorylated STAT3 expression. Cells were digested with trypsin-EDTA and counted. Using a multichannel pipette, 3,000 cells / 50 μL / well were evenly plated onto 96-well plates and incubated at 37°C with 5% CO2 until ready for use. To minimize edge effects, the outermost wells of the 96-well plate were not used for experiments and were filled with sterile PBS to minimize the effects of culture medium evaporation on cell growth and ensure experimental stability.

[0148] (2) Drug preparation

[0149] Drug concentration gradient design:

[0150] Drug A: STAT3 inhibitor. Napabucasin (BBI608) was used in this project.

[0151] Drug B: BCL-XL inhibitor, A1331852 was used in this project;

[0152] Negative control: The drug solvent is DMSO, and DMSO is used as a negative control in the implementation of this part of the project;

[0153] Drug A: In experiments with phosphorylated STAT3 high-expressing cell lines HGC-27, MKN-45, and AGS, a maximum concentration of drug was prepared at 4 times the actual concentration, and two concentrations were prepared according to a 4 / 5 gradient dilution. In experiments with phosphorylated STAT3 low-expressing cell lines SNU-216, NCI-N87, and GCIY, a maximum concentration of drug was prepared at 4 times the actual concentration, and two concentrations were prepared according to a 1 / 2 gradient dilution.

[0154] Drug B: In experiments with phosphorylated STAT3 high-expressing cell lines HGC-27, MKN-45, and AGS, a dose of 4 times the actual concentration was used to prepare the highest concentration, and then a 1 / 8 gradient dilution was used to prepare 5 concentrations. In experiments with phosphorylated STAT3 low-expressing cell lines SNU-216, NCI-N87, and GCIY, a dose of 4 times the actual concentration was used to prepare the highest concentration, and then a 1 / 2 gradient dilution was used to prepare 5 concentrations.

[0155] Negative control: Add DMSO at a volume corresponding to the highest drug concentration as a negative control.

[0156] Use a 1.8 mL 96-well plate to perform gradient dilutions of the above drugs.

[0157] (3) Drug treatment

[0158] Use a multichannel pipette to treat the cells plated in step (1) with the drug prepared in step (2).

[0159] Drug A concentration increasing direction: set to horizontal (row direction), 3 gradient concentrations, marked as C A (1-3);

[0160] Drug B concentration increasing direction: set to vertical (column direction), 6 gradient concentrations, marked as C B (1-6).

[0161] The settings of the 96-well plate are shown in Table 2:

[0162] Table 2 Drug concentration settings

[0163]

[0164] The cell activity was detected by combining BCLXL inhibitor and Napabucasin treatment for 72 hours.

[0165] (4) Cell activity detection: The cell activity detection method is as described above.

[0166] 2. Experimental results

[0167] (1) Figure 5 Shows the percentage heatmap and Bliss synergy score heatmap of cell viability of BCL-XL inhibitor A1331852 combined with STAT3 inhibitor Napabucasin in phosphorylated STAT3 high-expressing cell lines HGC27, MKN-45, AGS cell lines and phosphorylated STAT3 low-expressing cell lines SNU-216, NCI-N87, GCIY cell lines. Specific descriptions are as follows:

[0168] Vertical axis: represents the increasing concentration of drug A1331852;

[0169] Horizontal axis: represents the increasing concentration of the drug Napabucasin;

[0170] Cell activity percentage heat map white to red: indicates that the cell activity percentage decreases from 100 to 0.

[0171] Bliss synergy score heat map: blue indicates Bliss synergy score < 0, antagonistic effect, white indicates Bliss synergy score = 0, additive effect; red indicates Bliss synergy score > 0, synergistic effect.

[0172] The experimental results showed that BCL-XL inhibitors and STAT3 inhibitors had significant synergistic effects: the combination of A1331852 and Napabucasin showed strong synergistic effects in the phosphorylated STAT3 high-expressing cell lines HGC27, MKN-45, AGS cell lines and the phosphorylated STAT3 low-expressing cell lines SNU-216 and GCIY cell lines, and showed an additive effect in the phosphorylated STAT3 low-expressing NCI-N87 cells.

[0173] The above are only some embodiments of the present invention. For those skilled in the art, several modifications and improvements can be made without departing from the inventive concept of the present invention, which all fall within the scope of protection of the present invention.

Claims

1. Use of a BCL-XL inhibitor in combination with a HER2-targeted drug in the preparation of a medicament for the prevention and / or treatment of cancer; The BCL-XL inhibitor is selected from ABT263; The HER2 targeted drug is selected from Lapatinib or Trastuzumab; The cancer is selected from gastroesophageal adenocarcinoma or ovarian cancer.

2. The use according to claim 1, characterized in that When the cancer is gastroesophageal adenocarcinoma, the concentration ratio of ABT263 to Lapatinib is (0.04 μM~2.5 μM:62.5 nM~1000 nM) or (2.5 μM~10 μM:125 nM~1000 nM); the concentration ratio of ABT263 to Trastuzumab is (0.04 μM~2.5 μM:12.5 μg / mL~200 μg / mL) or (2.5 μM~10 μM:25 μg / mL~200 μg / mL); When the cancer is ovarian cancer, the concentration ratio of ABT263 to Lapatinib is (0.04μM~0.16μM:0.625μM~10μM) or (0.625μM~10μM:2.5μM~10μM); the concentration ratio of ABT263 to Trastuzumab is (0.04μM:12.5μg / mL~100μg / mL) or (25μM~10μM:25μg / mL~200μg / mL).

3. The use of a combination of a BCL-XL inhibitor and a HER2-targeted drug in the preparation of a reagent for inhibiting cancer cell activity or promoting cancer cell apoptosis in vitro; The BCL-XL inhibitor is selected from ABT263; The HER2 targeted drug is selected from Lapatinib or Trastuzumab; The cancer cells are selected from gastroesophageal adenocarcinoma cells or ovarian cancer cells; When the cancer is gastroesophageal adenocarcinoma, the concentration ratio of ABT263 to Lapatinib is (0.04 μM~2.5 μM:62.5 nM~1000 nM) or (2.5 μM~10 μM:125 nM~1000 nM); the concentration ratio of ABT263 to Trastuzumab is (0.04 μM~2.5 μM:12.5 μg / mL~200 μg / mL) or (2.5 μM~10 μM:25 μg / mL~200 μg / mL); When the cancer is ovarian cancer, the concentration ratio of ABT263 to Lapatinib is (0.04μM~0.16μM:0.625μM~10μM) or (0.625μM~10μM:2.5μM~10μM); the concentration ratio of ABT263 to Trastuzumab is (0.04μM:12.5μg / mL~100μg / mL) or (25μM~10μM:25μg / mL~200μg / mL).

4. A method for inhibiting cancer cell activity or promoting cancer cell apoptosis in vitro for non-therapeutic purposes, characterized in that: The method comprises: treating cancer cells with a BCL-XL inhibitor and a HER2-targeted drug in combination; The BCL-XL inhibitor is selected from ABT263; The HER2 targeted drug is selected from Lapatinib or Trastuzumab; The cancer cells are selected from gastroesophageal adenocarcinoma cells or ovarian cancer cells; When the cancer is gastroesophageal adenocarcinoma, the concentration ratio of ABT263 to Lapatinib is (0.04 μM~2.5 μM:62.5 nM~1000 nM) or (2.5 μM~10 μM:125 nM~1000 nM); the concentration ratio of ABT263 to Trastuzumab is (0.04 μM~2.5 μM:12.5 μg / mL~200 μg / mL) or (2.5 μM~10 μM:25 μg / mL~200 μg / mL); When the cancer is ovarian cancer, the concentration ratio of ABT263 to Lapatinib is (0.04μM~0.16μM:0.625μM~10μM) or (0.625μM~10μM:2.5μM~10μM); the concentration ratio of ABT263 to Trastuzumab is (0.04μM:12.5μg / mL~100μg / mL) or (25μM~10μM:25μg / mL~200μg / mL).

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