Combined drug for resisting tumor drug resistance, pharmaceutical composition and application

Abstract

The invention provides a combined drug for resisting tumor drug resistance, a pharmaceutical composition and application. The combined drug provided by the invention comprises an inhibitor of NADPH circulation related genes and an anti-tumor drug. The inhibitor of the NADPH circulation related gene is combined with the anti-tumor drug for use, so that the killing effect of the anti-tumor drug on drug-resistant cells can be remarkably enhanced. Besides, due to the addition of the inhibitor for the NADPH circulation related genes, the sensitivity of drug-resistant cells to drugs can be effectively improved, and meanwhile, the dosage of the inhibitor for the NADPH circulation related genes does not cause relatively large toxic damage to the cells when the inhibitor is independently used.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and specifically relates to an anti-tumor drug resistance combined drug, a pharmaceutical composition and a use. BACKGROUND

[0002] Anti-tumor drugs can act through various mechanisms. According to the mechanism classification, the drugs include drugs directly destroying DNA structure, such as alkylating agents and metal complexes (platinum); anti-metabolic drugs interfering with DNA synthesis, such as thymidylate synthetase inhibitors, dihydrofolate reductase inhibitors, purine analogs and pyrimidine analogs; topoisomerase inhibitors, free radical-mediated DNA damage agents, PARP inhibitors, small molecule kinase inhibitors, immune checkpoint inhibitors, epigenetic modulators, etc.

[0003] However, clinical studies have found that some patients do not respond to anti-tumor drugs, i.e., there is primary drug resistance; some tumor cells that are originally sensitive to drugs will gradually develop acquired drug resistance during chemotherapy, thereby leading to tumor progression and recurrence, and seriously affecting the effect of chemotherapy, which is also the main reason for the failure of chemotherapy.

[0004] Therefore, it is an urgent need in the clinic to develop a new method for treating tumors that can effectively reduce the drug resistance of anti-tumor drugs. SUMMARY

[0005] The present application aims to at least partially solve at least one of the problems in the prior art.

[0006] The present application is based on the following findings of the inventors:

[0007] NADPH (Nicotinamide Adenine Dinucleotide Phosphate) is one of the most important reducing power carriers in cells and plays a key role in various biochemical reactions. NADPH is mainly involved in anabolism and antioxidant defense. NADPH cycle-related genes include NADPH generation, NADH generation, NADH consumption, NADPH intermediates and various genes, which are involved in generation, utilization and regeneration, and the dynamic balance thereof is crucial for maintaining cell homeostasis. NADPH cycle has multiple source generation pathways and high metabolic flexibility, and has been proven to be highly related to diseases. Abnormal NADPH metabolism is closely related to cancer, neurodegenerative diseases, metabolic syndrome, cardiovascular diseases, etc.

[0008] In the previous study, the inventors successfully constructed a cell model with drug resistance to antitumor drugs by long-term exposure of cells to antitumor drugs. It was found that the NADPH metabolic level in cells with strong drug resistance was significantly higher by using a NADPH near-infrared fluorescent probe (for the synthesis process, refer to the literature: Guan et al. Sensor Actuat B-Chem, 2022, 373, 132694.; Guan et al. Chem. Commun., 2023, 59, 1617-1620.). Based on this finding, the present application proposes an innovative combination therapy method: by reducing the NADPH cycle, combined with antitumor drugs, so as to improve the drug resistance of antitumor drugs.

[0009] Therefore, in the first aspect of the present application, the present application proposes a combination drug for antitumor drug resistance. According to an embodiment of the present application, the combination drug comprises: an inhibitor of NADPH cycle-related genes and an antitumor drug. Through a large number of experiments, the inventors found that the combination of the inhibitor of NADPH cycle-related genes and the antitumor drug can significantly enhance the killing effect of the antitumor drug on drug-resistant cells. In addition, the addition of the inhibitor of NADPH cycle-related genes can effectively improve the sensitivity of drug-resistant cells to drugs, and the dose of the inhibitor of NADPH cycle-related genes used will not cause relatively large toxic damage to cells when used alone. Therefore, the combination drug of the present application not only improves the treatment effect, but also reduces the potential side effects, and provides a safe and effective treatment method for overcoming tumor drug resistance.

[0010] According to an embodiment of the present application, the combination drug can further comprise at least one of the following additional technical features:

[0011] According to an embodiment of the present application, the dose of the antitumor drug is an antitumor effective amount.

[0012] According to an embodiment of the present application, the dose of the inhibitor of NADPH cycle-related genes is a dose required to keep the cell growth or activity at 30%-90% when used alone.

[0013] According to an embodiment of the present application, the dose of the inhibitor of NADPH cycle-related genes is a dose required to keep the cell growth or activity at 70%-80% when used alone.

[0014] According to embodiments of the present application, the NADPH cycle-related genes include at least one of IDH2, IDH1, CBR1, G6PD, PGD, CBR3, FDXR, MTHFD1, MDH1, MDH2, DUS3L, MTHFD2, IDH3A, ALDH1B1, ALDH2, ALDH4A1, ALDH7A1, PHGDH.

[0015] According to embodiments of the present application, the PHGDH inhibitor includes at least one of NCT-502, NCT-503, CBR-5884, PHGDH-IN-2, PHGDH-IN-3, PHGDH-IN-4, PHGDH-IN-5, BI-4916, BI-4924, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0016] According to embodiments of the present application, the IDH1 inhibitor includes at least one of AGI-5198, IDH-305, GSK864, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0017] According to embodiments of the present application, the IDH2 inhibitor includes at least one of AG-221 (Enasidenib), AG-881 (Vorasidenib), AGI-12026, AGI-6780, IDH1 / 2-IN-1, GSK321, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0018] According to embodiments of the present application, the G6PD inhibitor includes at least one of RRx-001, G6PDi-1, Polydatin, 6-Aminonicotinamide, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0019] According to embodiments of the present application, the CBR1 inhibitor includes at least one of Miquelianin, Rutin, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0020] According to embodiments of the present application, the MTHFD1 inhibitor includes at least one of LY 345899, DS18561882, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0021] According to embodiments of the present application, the MDH1 inhibitor includes at least one of MDH1-IN-1, MDH1-IN-2, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0022] According to an embodiment of the present application, the MDH2 inhibitor comprises LW6, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0023] According to an embodiment of the present application, the MTHFD2 inhibitor comprises at least one of MTHFD2-IN1, MTHFD2-IN2, MTHFD2-IN3, MTHFD2-IN4, MTHFD2-IN5, DS44960156, TH9619, LY 345899, DS18561882, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0024] According to an embodiment of the present application, the ALDH1B1 inhibitor comprises IGUANA-1, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0025] According to an embodiment of the present application, the ALDH2 inhibitor comprises at least one of Alda-1, Alda-2, Daidzin, CVT-10216, 4-Hydroxynonenal, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0026] According to an embodiment of the present application, the anti-tumor drug comprises at least one of alkylating agents, anti-metabolites, plant alkaloids, antibiotics, small molecule kinase inhibitors, monoclonal antibody drugs, immune checkpoint inhibitors, epigenetic modulators, antibody conjugated drugs, metabolic targeting drugs, PARP inhibitors.

[0027] According to an embodiment of the present application, the anti-tumor drug comprises at least one of temozolomide, doxorubicin (Doxorubicin), cyclophosphamide, chlorambucil, carmustine, lomustine, cisplatin, oxaliplatin, carboplatin, 5-fluorouracil, carbinetrine, methotrexate, pemetrexed, gemcitabine, irinotecan, topotecan, etoposide, cytarabine, paclitaxel, docetaxel, bleomycin, dactinomycin, osimertinib, erlotinib, palbociclib, trastuzumab, rituximab, bevacizumab, vorinostat, azacitidine, olaparib, niraparib.

[0028] According to an embodiment of the present application, the inhibitor of NADPH cycle related genes is RRx-001, the anti-tumor drug is temozolomide, and the molar ratio of the RRx-001 to the temozolomide is (0.5-2):(1000-2500).

[0029] According to an embodiment of the present application, the molar ratio of the RRx-001 to the temozolomide is (1.3-2):(1050-2200).

[0030] According to an embodiment of the present application, the molar ratio of the RRx-001 to the temozolomide is (1.3-1.5):(1050-2200).

[0031] According to an embodiment of the present application, the inhibitor of the NADPH cycle-related gene is AG-221, the antitumor drug is temozolomide, and the molar ratio of the AG-221 to the temozolomide is (1-4):(70-200).

[0032] According to an embodiment of the present application, the molar ratio of the AG-221 to the temozolomide is (2-4):(70-200).

[0033] According to an embodiment of the present application, the molar ratio of the AG-221 to the temozolomide is (2.8-4):(140-200).

[0034] According to an embodiment of the present application, the inhibitor of the NADPH cycle-related gene is AGI-5198, the antitumor drug is doxorubicin, and the molar ratio of the AGI-5198 to the doxorubicin is (50-500):(0.5-5).

[0035] According to an embodiment of the present application, the molar ratio of the AGI-5198 to the doxorubicin is (100-200):(1-2).

[0036] According to an embodiment of the present application, the inhibitor of the NADPH cycle-related gene is RRx-001, the antitumor drug is doxorubicin, and the molar ratio of the RRx-001 to the doxorubicin is (0.5-5):(0.5-5).

[0037] According to an embodiment of the present application, the molar ratio of the RRx-001 to the doxorubicin is (0.5-2):(0.5-2).

[0038] According to an embodiment of the present application, the inhibitor of the NADPH cycle-related gene and the antitumor drug are used simultaneously, separately, or sequentially.

[0039] According to an embodiment of the present application, the inhibitor of the NADPH cycle-related gene and the antitumor drug are used simultaneously.

[0040] In a second aspect, the present application provides a pharmaceutical composition. According to embodiments of the present application, the pharmaceutical composition comprises an inhibitor of NADPH cycle-related genes and an antitumor drug as active ingredients, and a pharmaceutically acceptable carrier. Through a large number of experiments, the inventors have found that the combination of an inhibitor of NADPH cycle-related genes and an antitumor drug can significantly enhance the killing effect of the antitumor drug on drug-resistant cells. In addition, the addition of the inhibitor of NADPH cycle-related genes can effectively improve the sensitivity of drug-resistant cells to drugs, and the dosage of the inhibitor of NADPH cycle-related genes used does not cause significant toxic damage to cells when used alone. Therefore, the use of the pharmaceutical composition of the present application not only improves the therapeutic effect, but also reduces the potential side effects, and provides a safe and effective treatment method for overcoming tumor drug resistance.

[0041] In a third aspect, the present application provides the use of the combination drug of the first aspect or the pharmaceutical composition of the second aspect in the preparation of an antitumor drug. As described above, the combination drug and the pharmaceutical composition of the present application can improve the sensitivity of drug-resistant cells to drugs, thereby enhancing the killing effect of the antitumor drug. Therefore, the preparation of the combination drug as a drug can effectively improve the antitumor effect of the antitumor drug.

[0042] According to embodiments of the present application, the tumor comprises at least one of brain glioma, glioblastoma, neuroblastoma, breast cancer, ovarian cancer, colorectal cancer, gastric cancer, lymphoma, sarcoma, testicular cancer, lung cancer, melanoma, and leukemia.

[0043] Additional aspects and advantages of the present application will be in part apparent and in part pointed out hereinafter. BRIEF DESCRIPTION OF DRAWINGS

[0044] The above and / or additional aspects and advantages of the present application will become apparent and be readily appreciated from the following description, including the accompanying drawings, wherein:

[0045] Figure 1 is the synergistic index of temozolomide and RRx-001 according to embodiment 1 of the present application in the treatment of neuroblastoma SH-SY5Y drug-resistant cells when used in combination;

[0046] Figure 2 is the NADPH metabolic change of neuroblastoma SH-SY5Y drug-resistant cells observed using an NADPH probe during the combined treatment of temozolomide and RRx-001 according to embodiment 1 of the present application and the treatment of temozolomide alone; wherein,

[0047] (a) is flow cytometry data comparing NADPH probe uptake in SH-SY5Y drug resistant cells during temozolomide alone treatment and temozolomide and RRx-001 combination treatment;

[0048] (b) is quantification of flow cytometry data on probe uptake;

[0049] (c-d) are fluorescence confocal microscopy observations comparing NADPH probe uptake in SH-SY5Y drug resistant cells during temozolomide (TMZ) alone treatment and temozolomide and RRx-001 combination treatment;

[0050] (e) is quantification of fluorescence confocal microscopy observations on probe uptake;

[0051] Figure 3 is the change in tumor volume in mice bearing SH-SY5Y drug resistant neuroblastoma subcutaneous tumors during temozolomide and RRx-001 combination treatment and temozolomide alone treatment according to Example 1 of the present application;

[0052] Figure 4 is the difference in behavioral changes in mice after temozolomide and RRx-001 combination treatment and temozolomide alone treatment according to Example 1 of the present application; wherein,

[0053] (a-b) are representative behavioral traces of mice in the open field test during temozolomide alone treatment and temozolomide and RRx-001 combination treatment;

[0054] (c) is quantification of total distance traveled in the open field test for mice during temozolomide alone treatment and temozolomide and RRx-001 combination treatment;

[0055] (d) is quantification of time spent in the center zone in the open field test for mice during temozolomide alone treatment and temozolomide and RRx-001 combination treatment;

[0056] (e-f) are representative behavioral traces of mice in the elevated plus maze test during temozolomide alone treatment and temozolomide and RRx-001 combination treatment;

[0057] (g) is quantification of total distance traveled in the elevated plus maze test for mice during temozolomide alone treatment and temozolomide and RRx-001 combination treatment;

[0058] (h) is quantification of number of open arm explorations in the elevated plus maze test for mice during temozolomide alone treatment and temozolomide and RRx-001 combination treatment;

[0059] Figure 5Synergy index of temozolomide and AG-221 according to Example 2 of the present application when used in combination in the treatment of neuroblastoma SH-SY5Y resistant cells;

[0060] Figure 6 Therapeutic effect of doxorubicin (DOX) and AGI-5198 according to Example 3 of the present application when administered alone or in combination in the treatment of breast cancer MCF-7 cells;

[0061] Figure 7 Therapeutic effect of doxorubicin and RRx-001 according to Example 4 of the present application when administered alone or in combination in the treatment of breast cancer MCF-7 cells. DETAILED DESCRIPTION

[0062] The embodiments of the present application are described in detail below, with the purpose of explaining the present application, and not to be understood as limiting the present application.

[0063] Reference herein to "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment can be included in at least one embodiment of the application. The appearances of the phrase that an embodiment in various places in the specification are not necessarily all referring to the same embodiment, nor are they necessarily mutually exclusive of one another. It is expressly understood that the embodiments described herein are merely examples from a

[0064] For the sake of clarity, only some numerical ranges are specifically disclosed herein. However, any lower limit can be combined with any upper limit to form a range not explicitly recited; and any lower limit can be combined with any other lower limit to form a range not explicitly recited, as well as any upper limit can be combined with any other upper limit to form a range not explicitly recited. Further, each individual disclosed point or single numerical value can itself be combined as a lower limit or upper limit with any other point or single numerical value or with other lower limits or upper limits to form a range not explicitly recited.

[0065] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs; the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the present application; the description and drawings of the present application together with the attached figures are incorporated herein and constitute a part of the detailed description of the application, and the terms "comprising", "having", "including" and any variations thereof used herein are intended to cover not exclusive inclusion, but are intended to cover both "including" and "consisting of".

[0066] In this document, the terms "comprise" or "comprising" are open- ended, that is, they mean "including, but not limited to", they do not exclude other moieties, additives, components, integers or steps.

[0067] In the present text, the terms "optionally", "optional" or "option" generally mean that the event or circumstance described subsequently can, but need not, occur, and the description includes situations in which the event or circumstance occurs, as well as situations in which it does not.

[0068] In the present text, the term "refractory to (a treatment)" in relation to a cancer means a cancer that is unresponsive to a particular anti-neoplastic drug treatment, or that relapses after being responsive to a particular anti-neoplastic drug treatment. Thus, for example, a neuroblastoma that is refractory to temozolomide means that a treatment regimen comprising, but not limited to, temozolomide is unresponsive to treat the neuroblastoma, or that the neuroblastoma relapses after being responsive to the treatment regimen comprising, but not limited to, temozolomide.

[0069] To detect or determine a refractory cancer, a patient receiving chemotherapy can be carefully monitored for signs of drug resistance, non-response to treatment, or cancer relapse. This can be done by monitoring the patient's cancer for response to the chemotherapy treatment. The patient's response, non-response, or cancer relapse to the initial treatment can be detected using any suitable method in the art. For example, this can be done by assessing the size and number of tumors. If the tumor size increases or the number of tumors increases, this indicates that the tumor is unresponsive to the chemotherapy, or that the tumor is showing signs of relapse. A refractory cancer can be determined according to the detailed description of the "RECIST" criteria published by Therasse et al (Therasse et al, J. Natl. Cancer Inst., 92:205-216 (2000)).

[0070] In the present text, the term "pharmaceutically acceptable salt" means a salt that does not have significant deleterious effects on the patients to whom it is administered and that is useful as a pharmaceutical composition. A compound having an acidic substituent can form a salt by reacting with a base. Examples of such salts include alkali metal salts such as sodium salts, potassium salts, lithium salts; alkaline earth metal salts such as calcium salts, magnesium salts; metal salts such as aluminum salts, iron salts; inorganic salts such as ammonium salts; amine salts such as t-butylamine salts, t-octylamine salts, dibenzylamine salts, morpholine salts, glucosamine salts, phenylglycine alkyl ester salts, ethylenediamine salts, N-methylglucosamine salts, guanidine salts, diethylamine salts, triethylamine salts, dicyclohexylamine salts, N,N'-dibenzylethylenediamine salts, chloroprocaine salts, procaine salts, diolamine salts, N-benzylphenethylamine salts, piperazine salts, tetramethylammonium salts, tris(hydroxymethyl)aminomethane salts; and amino acid salts such as glycine salts, lysine salts, arginine salts, ornithine salts, glutamic acid salts, aspartic acid salts, but not limited to these.

[0071] Compounds having basic substituents can form salts by reaction with acids. Examples of such salts are hydrohalides, such as hydrofluorides, hydrochlorides, hydrobromides, hydroiodides; salts of inorganic acids, such as nitrates, perchlorates, sulfates, phosphates; salts of C1-C6 alkylsulfonic acids, such as methanesulfonates, trifluoromethanesulfonates, ethanesulfonates; salts of arylsulfonic acids, such as benzenesulfonates, p-toluenesulfonates; salts of organic acids, such as acetates, malates, fumarates, succinates, citrates, ascorbates, tartrates, oxalates, adipates, maleates; and salts of amino acids, such as glycine, lysine, arginine, ornithine, glutamic acid, aspartic acid.

[0072] The present application also encompasses salts formed when an acidic proton present in the parent compound either is replaced by a metal ion, e.g., an alkali metal ion, an alkaline earth metal ion, or an ammonium ion; or coordinates with an organic base, such as ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, or the like. In salt forms, it is understood that the ratio of the compound to the cation or anion of the salt can be 1:1, or any ratio other than 1:1, such as 3:1, 2:1, 1:2, or 1:3.

[0073] The compounds of the present application can also be prepared as esters, for example, pharmaceutically acceptable esters. For example, a carboxylic acid functionality in a compound can be converted to its corresponding ester, for example, a methyl ester, an ethyl ester, or other ester. Also, an alcohol group in a compound can be converted to its corresponding ester, for example, an acetate, a propionate, or other ester.

[0074] In the present context, the term "solvate" refers to an association or complex of one or more solvent molecules with a compound of the present application. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, aminoethanol. The term "hydrate" refers to an association or complex of water molecules with a compound of the present application.

[0075] In the present context, the term "treatment" when referring to a tumor means having a therapeutic effect on the tumor, alleviating one or more symptoms of the tumor, altering the progression of the tumor, eradicating the tumor, reducing the size of the tumor, slowing or inhibiting the growth of the tumor, delaying or minimizing one or more symptoms associated with the tumor, reducing the malignancy of the tumor, or inducing dormancy of the tumor, or alleviating or minimizing one or more side effects associated with another therapy administered or applied to treat the tumor. It includes not only complete recovery, but also inhibition of progression of cancer (inhibition of proliferation of cancerous tissue, reduction of cancerous tissue, etc.), inhibition of occurrence of cancer (inhibition of secondary cancer occurrence, inhibition of recurrence of cancer, etc.), alleviation of symptoms associated with cancer.

[0076] The present application provides a combination drug for resisting tumor drug resistance, a pharmaceutical composition and use thereof, which will be described in detail below.

[0077] Combination drug

[0078] In a first aspect, the present application provides a combination drug for resisting tumor drug resistance. According to an embodiment of the present application, the combination drug comprises an inhibitor of NADPH cycle-related genes and an anti-tumor drug. Through a large number of experiments, the inventors found that the combination of the inhibitor of NADPH cycle-related genes and the anti-tumor drug can significantly enhance the killing effect of the anti-tumor drug on drug-resistant cells. In addition, the addition of the inhibitor of NADPH cycle-related genes can effectively improve the sensitivity of drug-resistant cells to drugs, and the dose of the inhibitor of NADPH cycle-related genes used will not cause relatively large toxic damage to cells when used alone. Therefore, the combination drug of the present application not only improves the therapeutic effect, but also reduces the potential side effects, and provides a safe and effective treatment method for overcoming tumor drug resistance.

[0079] In some embodiments of the present application, the dose of the anti-tumor drug is an anti-tumor effective amount. Wherein, the "anti-tumor effective amount" refers to any amount that protects the subject from slowing down the development of tumors, reducing tumor complications, reducing tumor severity, reducing tumor recurrence, reducing tumor duration, etc. by using the anti-tumor drug alone. The ability of a therapeutic agent to promote disease regression can be evaluated using a variety of methods known to the skilled practitioner, such as a doctor, a researcher, etc., such as in human subjects during clinical trials, in animal model systems predictive of efficacy in humans, or by assaying the activity of the agent in in vitro assays.

[0080] In order to reduce the NADPH metabolism in tumors, improve the killing effect of anti-tumor drugs on drug-resistant tumors, and reduce the impact on normal cell function, the present application sets the dose of the inhibitor of NADPH cycle-related genes to be the dose required to maintain cell growth or activity at 30%-90% when acting alone, or to be the dose required to maintain cell growth or activity at 70%-80% when acting alone. For example, it can be 30%, 40%, 50%, 60%, 70%, 80%, 90%. This dose ensures that while effectively inhibiting tumor cell growth and activity, normal cells are maximally protected from damage. When the inhibitor of NADPH cycle-related genes is used in combination with the anti-tumor drug, this dose setting not only enhances the efficacy of the anti-tumor drug, but also avoids additional toxic damage to normal cells, thereby achieving the best balance between therapeutic effect and safety.

[0081] In some embodiments of the present application, the "anti-tumor drug" refers to various types of compounds used for tumor treatment, and the specific type is not particularly limited, for example, the following several categories can be covered: alkylating agents, antimetabolites, plant alkaloids, antibiotic drugs, small molecule kinase inhibitors, monoclonal antibody drugs, immune checkpoint inhibitors, epigenetic modulators, antibody conjugate drugs, metabolic targeting drugs, PARP inhibitors, etc. Exemplarily, it can be any one of the following: temozolomide, doxorubicin (doxorubicin), cyclophosphamide, chlorambucil, carmustine, lomustine, cisplatin, oxaliplatin, carboplatin, 5-fluorouracil, carbinetabin, methotrexate, pemetrexed, gemcitabine, irinotecan, topotecan, etoposide, cytarabine, paclitaxel, docetaxel, bleomycin, dactinomycin, osimertinib, erlotinib, palbociclib, trastuzumab, rituximab, bevacizumab, vorinostat, azacitidine, olaparib, niraparib, etc. It should be noted that there are many types of anti-tumor drugs, and their efficacy and safety can be evaluated by methods such as clinical trials, pharmacodynamics studies, etc. recognized by those skilled in the art.

[0082] In some embodiments of the present application, the "NADPH cycle inhibitor" refers to any compound that can reduce the intracellular NADPH or NADH content by inhibiting the activity of NADPH cycle-related genes. These inhibitors are not particularly limited as long as they can achieve the above function. Specifically, they can target related genes involved in NADPH generation, NADH generation, NADH consumption, NADPH metabolism, and NADPH intermediates in the NADPH metabolic cycle. These genes include but are not limited to the following: IDH2, IDH1, CBR1, G6PD, PGD, CBR3, FDXR, MTHFD1, MDH1, MDH2, DUS3L, MTHFD2, IDH3A, ALDH1B1, ALDH2, ALDH4A1, ALDH7A1, PHGDH. It should be noted that the effect of the inhibitor on the reduction of NADPH or NADH content can be evaluated and verified by experimental methods recognized by those skilled in the art.

[0083] As the "NADPH cycle inhibitor" of the present application, for example, the following substances can be exemplified:

[0084] (i) at least one of PHGDH inhibitors NCT-502, NCT-503, CBR-5884, PHGDH-IN-2, PHGDH-IN-3, PHGDH-IN-4, PHGDH-IN-5, BI-4916, BI-4924, or a pharmaceutically acceptable salt, ester or solvate thereof;

[0085] (ii) at least one of IDH1 inhibitors AGI-5198, IDH-305, GSK864, or a pharmaceutically acceptable salt, ester, or solvate thereof;

[0086] (iii) at least one of IDH2 inhibitors AG-221 (Enasidenib), AG-881 (Vorasidenib), AGI-12026, AGI-6780, IDH1 / 2-IN-1, GSK321, or a pharmaceutically acceptable salt, ester, or solvate thereof;

[0087] (iv) at least one of G6PD inhibitors RRx-001, G6PDi-1, Polydatin, 6-Aminonicotinamide, or a pharmaceutically acceptable salt, ester, or solvate thereof;

[0088] (v) at least one of CBR1 inhibitors Miquelianin, Rutin, or a pharmaceutically acceptable salt, ester, or solvate thereof;

[0089] (vi) at least one of MTHFD1 inhibitors LY 345899, DS18561882, or a pharmaceutically acceptable salt, ester, or solvate thereof;

[0090] (vii) at least one of MDH1 inhibitors MDH1-IN-1, MDH1-IN-2, or a pharmaceutically acceptable salt, ester, or solvate thereof;

[0091] (viii) at least one of MDH2 inhibitors LW6, or a pharmaceutically acceptable salt, ester, or solvate thereof;

[0092] (ix) at least one of MTHFD2 inhibitors MTHFD2-IN1, MTHFD2-IN2, MTHFD2-IN3, MTHFD2-IN4, MTHFD2-IN5, DS44960156, TH9619, LY 345899, DS18561882, or a pharmaceutically acceptable salt, ester, or solvate thereof;

[0093] (x) at least one of ALDH1B1 inhibitors IGUANA-1, or a pharmaceutically acceptable salt, ester, or solvate thereof;

[0094] (xi) at least one of ALDH2 inhibitors Alda-1, Alda-2, Daidzin, CVT-10216, 4-Hydroxynonenal, or a pharmaceutically acceptable salt, ester, or solvate thereof.

[0095] In some embodiments of the present application, the inhibitor of NADPH cycle related genes is RRx-001, the antitumor drug is temozolomide, and the molar ratio of RRx-001 to temozolomide is (0.5-2):(1000-2500). In some embodiments of the present application, the molar ratio of RRx-001 to temozolomide is (1.3-2):(1050-2200). In some embodiments of the present application, the molar ratio of RRx-001 to temozolomide is (1.3-1.5):(1050-2200). Thus, by setting the molar ratio of RRx-001 to temozolomide in the above range, the killing effect of temozolomide on drug-resistant tumor cells can be significantly enhanced.

[0096] In some embodiments of the present application, the inhibitor of NADPH cycle related genes is AG-221, the antitumor drug is temozolomide, and the molar ratio of AG-221 to temozolomide is (1-4):(70-200). In some embodiments of the present application, the molar ratio of AG-221 to temozolomide is (2-4):(70-200). In some embodiments of the present application, the molar ratio of AG-221 to temozolomide is (2.8-4):(140-200). Thus, by setting the molar ratio of AG-221 to temozolomide in the above range, the killing effect of temozolomide on drug-resistant tumor cells can be significantly enhanced.

[0097] In some embodiments of the present application, the inhibitor of NADPH cycle related genes is AGI-5198, the antitumor drug is doxorubicin, and the molar ratio of AGI-5198 to doxorubicin is (50-500):(0.5-5). In some embodiments of the present application, the molar ratio of AGI-5198 to doxorubicin is (100-200):(1-2). Thus, by setting the molar ratio of AGI-5198 to doxorubicin in the above range, the killing effect of doxorubicin on drug-resistant tumor cells can be significantly enhanced.

[0098] In some embodiments of the present application, the inhibitor of NADPH cycle-related genes is RRx-001, the antitumor drug is doxorubicin, and the molar ratio of RRx-001 to doxorubicin is (0.5-5):(0.5-5). In some embodiments of the present application, the molar ratio of RRx-001 to doxorubicin is (0.5-2):(0.5-2). In this way, by setting the molar ratio of RRx-001 to doxorubicin within the above range, the killing effect of doxorubicin on drug-resistant tumor cells can be significantly enhanced.

[0099] The combination drug regimen used in the present application can administer the inhibitor of NADPH cycle-related genes and the antitumor drug at the same time, or in a staggered administration regimen, i.e., the inhibitor of NADPH cycle-related genes can be administered at a different time from the antitumor drug. The time interval between the administration of the two drugs ranges from a few minutes, a few hours, a few days, a few weeks, or even longer. Therefore, the term combination drug does not necessarily mean simultaneous administration or administration as a single agent, but the components are administered during the required treatment period. The drugs can also be administered by different routes. For example, in the combination drug of the inhibitor of NADPH cycle-related genes and the antitumor drug, it is expected that the inhibitor of NADPH cycle-related genes can be administered orally or parenterally; while the antitumor drug can be administered parenterally, orally, or by other acceptable means. These combination drugs can be administered daily, weekly, or even monthly.

[0100] Pharmaceutical composition

[0101] In a second aspect of the present application, a pharmaceutical composition is provided. According to embodiments of the present application, the pharmaceutical composition comprises an inhibitor of NADPH cycle-related genes and an antitumor drug as active ingredients, and a pharmaceutically acceptable carrier. Through a large number of experiments, the inventors have found that the combination of an inhibitor of NADPH cycle-related genes and an antitumor drug can significantly enhance the killing effect of the antitumor drug on drug-resistant cells. In addition, the addition of an inhibitor of NADPH cycle-related genes can effectively improve the sensitivity of drug-resistant cells to drugs. Therefore, the use of the pharmaceutical composition of the present application not only improves the therapeutic effect, but also reduces the potential side effects, providing a safe and effective treatment method for overcoming tumor drug resistance.

[0102] In the present application, when the NADPH cycle inhibitor and / or the antitumor drug is prepared into a pharmaceutical composition, as a pharmaceutically acceptable carrier used, for example, sterile water, physiological saline, vegetable oil, solvent, base, emulsifier, suspending agent, surfactant, stabilizer, flavoring agent, aromatic agent, preservative, binding agent, diluent, isotonic agent, painless agent, bulking agent, disintegrating agent, buffer, coating agent, lubricant, coloring agent, sweetening agent, thickening agent, corrigent, solubilizing agent or other additives, etc. can be exemplified, but are not limited to these. The pharmaceutical composition of the present application can be prepared into various forms such as tablets, powders, granules, capsules, liquid preparations, etc. according to the therapeutic purpose, etc. In addition, for example, it can also be administered in the form of a liposome delivery system. The above-mentioned auxiliary moieties (for example, antibodies, ligands, etc.) which enhance the therapeutic useful properties can also be added to the liposome.

[0103] In the present application, when the NADPH cycle inhibitor and / or the antitumor drug is prepared into a pharmaceutical composition, the preparation method can be performed by mixing, dissolving, granulating, coating, milling, emulsifying, encapsulating, entrapping or lyophilizing processes, etc. according to the conventional preparation methods in the art. The pharmaceutical composition can be formulated in a conventional manner using one or more pharmaceutically acceptable carriers (including excipients and / or auxiliaries) which facilitate processing of the active ingredient into preparations which are suitable for pharmaceutical use. Of course, the specific formulation will be determined by the selected route of administration.

[0104] The pharmaceutical composition of the present application can be in the form of a sterile injectable preparation, for example, as a sterile injectable aqueous or oleaginous suspension. This suspension can be formulated according to techniques known in the art using suitable dispersing or wetting agents (such as sulfobutyl-β-cyclodextrin, Tween) and suspending agents. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be employed are mannitol, water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil can be employed including synthetic mono- or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions can also contain long-chain alcohol diluents or dispersants, or carboxymethyl cellulose or similar dispersing agents, which are commonly used in the formulation of pharmaceutically acceptable dosage forms such as emulsions and / or suspensions. Other commonly used surfactants, such as Tween or Span and / or other similar emulsifying agents or bioavailability enhancers, which are commonly used in manufacturing pharmaceutically acceptable solid, liquid, or other dosage forms, can also be used for formulation purposes.

[0105] The pharmaceutical composition of the present application can also be used as a reagent. In the present application, when the inhibitor of the NADPH cycle-related gene and / or the antitumor drug are prepared as a reagent, other components permitted to be used as a reagent such as sterilized water, physiological saline, a buffer, a preservative, and the like can be included as necessary. As to the reagent, a subject of interest (e.g., a cell and a fraction thereof, a tissue, an experimental animal, and the like) is administered with an amount of administration of interest, for example, an amount capable of inhibiting the activity of the NADPH metabolic cycle enzyme, inhibiting the production of NADPH, inhibiting the production of NADH, promoting the consumption of NADPH, promoting the consumption of NADH, and inhibiting the proliferation of a tumor.

[0106] Uses and methods

[0107] In a third aspect of the present application, the present application provides use of the combination drug of the first aspect or the pharmaceutical composition of the second aspect in the manufacture of a medicament for drug resistance against a tumor. As described above, the combination drug and the pharmaceutical composition of the present application can improve the sensitivity of drug-resistant cells to a drug, thereby enhancing the killing effect of an antitumor drug, and thus, when prepared as a medicament, can effectively improve the antitumor effect of the antitumor drug.

[0108] In some embodiments of the present application, various combinations of the inhibitor of the NADPH cycle-related gene and the antitumor drug act synergistically and can be used as a medicament for drug resistance against a tumor. More specifically, the medicament can be used for the treatment of brain glioma, glioblastoma, neuroblastoma, breast cancer, renal cancer, soft tissue sarcoma, sarcoma, testicular cancer, pulmonary neuroendocrine tumor, cervical cancer, uterine cancer, head and neck cancer, glioma, non-small cell lung cancer, prostate cancer, pancreatic cancer, lymphoma, melanoma, small cell lung cancer, ovarian cancer, colon cancer, esophageal cancer, gastric cancer, leukemia, colorectal cancer, and unknown primary cancer.

[0109] In some embodiments of the present application, the tumor includes at least one of brain glioma, glioblastoma, neuroblastoma, breast cancer, ovarian cancer, colorectal cancer, gastric cancer, lymphoma, sarcoma, testicular cancer, lung cancer, melanoma, and leukemia.

[0110] In a fourth aspect of the present application, the present application provides a method of treating a drug-resistant tumor. According to an embodiment of the present application, the method includes administering to a subject an effective amount of the combination drug of the first aspect or the pharmaceutical composition of the second aspect.

[0111] In the present application, the subject treated by the methods of the present application is desirably a human subject, although it should be understood that the methods described herein are effective for all vertebrate species, which are intended to be encompassed by the term "subject." Thus, a "subject" can include a human subject for medical purposes, e.g., for treatment of an existing condition or disease or for prophylactic treatment to prevent the onset of a condition or disease, or an animal (non-human) subject for medical, veterinary, or developmental purposes. Suitable animal subjects include mammals, including but not limited to primates, e.g., humans, monkeys, apes, etc.; bovids, e.g., cattle, oxen, etc.; ovids, e.g., sheep, etc.; caprids, e.g., goats, etc.; suids, e.g., pigs, hogs, etc.; equids, e.g., horses, donkeys, zebras, etc.; felines, including wild cats and domestic cats; canids, including dogs; lagomorphs, including rabbits, hares, etc.; and rodents, including mice, rats, etc. An animal can be a transgenic animal. In some embodiments, the subject is a human, including but not limited to fetal, neonatal, infant, juvenile, and adult subjects. Further, a "subject" can include a patient having or suspected of having a condition or disease. Thus, the terms "subject" and "patient" are used interchangeably herein. In some embodiments, the subject is a human. In other embodiments, the subject is a non-human.

[0112] In the present application, administration to a subject can be oral administration or non-oral administration. As non-oral administration, various routes are contemplated, such as, for example, pulmonary, rectal, parenteral, transdermal, subcutaneous, intravenous, intra-arterial, intramuscular, intraperitoneal, inhalation, buccal, sublingual, intrapleural, intrathecal, intranasal, and the like. Dosage forms for topical or transdermal administration of a compound of the present application include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. In some embodiments, the active compound is mixed under sterile conditions with a pharmaceutically acceptable carrier, and any needed preservatives, buffers, or propellants.

[0113] In the present application, the amount of administration is not particularly limited as long as it is an amount effective for the treatment of the target disease, and can be appropriately selected depending on the age, body weight, symptoms, health status, progression status of the disease, and the like of the patient. The frequency of administration is also not particularly limited, and can be appropriately selected depending on the purpose, for example, the amount of administration per day can be set to administration once a day, or can be divided into multiple times for administration. When the medicament of the present application is administered to humans, the range of the amount of administration of each effective ingredient is generally about 0.01 mg / kg body weight to about 500 mg / kg body weight per day, and preferably about 0.1 mg / kg body weight to about 100 mg / kg body weight per day. When administered to humans, administration is preferably once a day, or divided into 2 to 4 times for administration, and preferably repeated at appropriate intervals.

[0114] Examples of the present application are described in detail below. The examples described below are illustrative, and are intended to explain the present application, and are not to be construed as limiting the present application. In the examples, specific techniques or conditions not otherwise described are performed according to techniques or conditions described in the literature in the field or according to product instructions. Reagents or instruments not otherwise described are all conventional products that can be obtained commercially.

[0115] Example 1: RRx-001 combined with temozolomide to improve neuroblastoma drug resistance

[0116] 1. Cell experiments

[0117] By culturing SH-SY5Y cells using a culture medium containing temozolomide, and gradually increasing to a culture medium containing 203 μM temozolomide, and stably culturing for more than 2 years, the change in IC 50 value was verified using the CCK-8 method and the CTG method, and it was confirmed that the IC 50 value of the cells was increased by 7 times compared to the drug-resistant SH-SY5Y cells, and at least 5 times higher than the currently recognized drug-resistant cells in IC 50 value, thereby obtaining SH-SY5Y cells with stable drug resistance to temozolomide.

[0118] The G6PD inhibitor RRx-001 was combined with temozolomide on SH-SY5Y drug-resistant cells, and the combination index (CI) of different concentration combinations was calculated by the Bliss independence model to evaluate the overall combination therapy effect. The experimental results are as follows Figure 1As shown, the average synergy index of RRx-001 combined with temozolomide was 16.59, indicating a significant synergistic effect between the two. Specifically, when the concentration of RRx-001 was 1.36 μM and the concentration of temozolomide was 1459.46 μM, the combined treatment was most effective and the synergistic effect was most significant, indicating that the combination of RRx-001 and temozolomide can effectively enhance the treatment effect on drug-resistant cells.

[0119] To investigate the effect of G6PD inhibitor RRx-001 combined with temozolomide on NADPH metabolism in SH-SY5Y drug-resistant cells, SH-SY5Y drug-resistant cells were treated with temozolomide alone (203 μM temozolomide) and combined with temozolomide (203 μM temozolomide and 0.5 μM RRx-001), respectively. After 48 h of treatment, the drug-resistant cells were incubated with NADPH fluorescent probes for 3 h. Then, the cells were observed using flow cytometry and fluorescence confocal microscopy. The experimental results are shown in Figure 2 As shown, the results showed that the level of NADPH metabolism decreased significantly during the combined treatment, and the combined treatment had a more obvious inhibitory effect on NADPH metabolism in drug-resistant cells compared to temozolomide alone. This finding indicates that the combination of RRx-001 and temozolomide may enhance the killing effect on drug-resistant cells by inhibiting NADPH metabolism.

[0120] 2. Animal treatment experiment

[0121] To evaluate the anti-tumor effect of G6PD inhibitor RRx-001 combined with temozolomide on SH-SY5Y drug-resistant cells, a subcutaneous tumor model was constructed in SCID mice. SH-SY5Y drug-resistant cells were implanted subcutaneously into the right hind leg of SCID mice. After the tumor formed, the long diameter and short diameter of the tumor were measured regularly using a vernier caliper, and the tumor volume (unit: mm 2 ) was calculated by the formula (long diameter) x (short diameter) 3 / 2 to dynamically monitor the proliferation of the tumor.

[0122] On the 18th day after subcutaneous tumor implantation, the SCID mice were randomly divided into two groups, each receiving a different treatment regimen: one group received temozolomide (50 mg / kg) treatment only, and the other group received temozolomide (50 mg / kg) combined with RRx-001 (5 mg / kg) treatment. During the treatment period, the test compounds were administered by intraperitoneal injection in a "5 days of administration, 2 days of rest" cycle for 2 weeks.

[0123] After the treatment ended, the change in tumor volume of the two groups of mice within 2 weeks after grouping was compared. The experimental results are shown in Figure 3As shown, the results showed that the treatment group using G6PD inhibitor RRx-001 in combination with temozolomide exhibited stronger tumor proliferation inhibition effect compared with the group using temozolomide alone. This indicates that RRx-001 in combination with temozolomide can significantly enhance the treatment effect on drug-resistant tumors.

[0124] 3. Animal behavior experiment

[0125] Subsequently, the mice treated with temozolomide (50 mg / kg) and temozolomide (50 mg / kg) in combination with RRx-001 (5 mg / kg) for two weeks were subjected to open field test and elevated plus maze test, respectively. In the open field test, the mice were placed in the center of the field and allowed to freely explore for 20 minutes, and the results are shown in Figure 4 As shown, no significant changes in the movement distance, movement speed and time spent in the center of the mice in the single treatment group and the combination treatment group were observed, and the mice in the combination treatment group slightly increased. In the elevated plus maze test, the mice were placed on the central platform and allowed to freely explore for 10 minutes. No significant changes in the movement distance, movement speed, frequency of entering the open arm and time spent in the open arm of the mice in the single treatment group and the combination treatment group were observed, and the mice in the combination treatment group slightly increased. These results indicate that the combination treatment method used in the present application does not cause abnormal behavior in mice and does not cause anxiety-like behavior in mice, proving the potential application value of the combination treatment regimen.

[0126] Example 2: AG-221 in combination with temozolomide improves neuroblastoma drug resistance

[0127] By culturing SH-SY5Y cells using temozolomide-containing medium and gradually increasing the medium containing 203 μM temozolomide for more than 2 years, the change in IC 50 value was verified by CCK-8 method and CTG method, and it was confirmed that the IC 50 value of the cells was increased by 7 times compared with the drug-resistant SH-SY5Y cells, and at least 5 times higher than the currently recognized drug-resistant cells, thereby obtaining SH-SY5Y cells with stable drug resistance to temozolomide. 50

[0128] The IDH2 inhibitor AG-221 was combined with temozolomide on SH-SY5Y drug-resistant cells, and the combination treatment index of different concentration combinations was calculated by Bliss independence model, and the overall combination treatment effect was evaluated. The experimental results are shown in Figure 5 ​As shown in Table 1, the average synergy index of AG-221 and temozolomide is 11.18, indicating that there is a synergistic effect between the two. Specifically, when the concentration of AG-221 is 36.08 μM and the concentration of temozolomide is 1824.32 μM, the combined treatment effect is best and the synergistic effect is most significant, which indicates that the combination of AG-221 and temozolomide can effectively enhance the therapeutic effect on drug-resistant cells.

[0129] Example 3: AGI-5198 combined with doxorubicin improves the treatment of breast cancer drug resistance

[0130] According to the results of the previous experiments, the effective concentration range of doxorubicin and AGI-5198 was selected for treatment, and the concentration of AGI-5198 that can keep the cell activity above 80% is below 200 μM.

[0131] In order to evaluate the combined treatment effect of IDH1 inhibitor AGI-5198 and doxorubicin on breast cancer MCF-7 cells, MCF-7 cells were randomly divided into five groups, one of which was a control group, one of which was treated with only 1.5 μM doxorubicin, one of which was treated with 100 μM AGI-5198, one of which was treated with 100 μM AGI-5198 and 1.5 μM doxorubicin, and one of which was treated with 200 μM AGI-5198 and 1.5 μM doxorubicin. After 48 h of treatment, the activity of the cells was detected using the CCK-8 experiment, and the therapeutic effect of the drugs was evaluated by calculating the inhibition rate.

[0132] The experimental results are shown in Table 2. Figure 6 As shown in Table 2, the results show that the inhibition rate of the combined drug group is significantly higher than that of the single drug group, indicating that the combination of IDH1 inhibitor AGI-5198 and doxorubicin can significantly enhance the inhibition effect on MCF-7 cells. This result indicates that combined treatment can more effectively inhibit cell proliferation and improve the therapeutic effect of drugs.

[0133] Example 4: RRx-001 combined with doxorubicin improves the treatment of breast cancer drug resistance

[0134] According to the results of the previous experiments, the effective concentration range of doxorubicin and RRx-001 was selected for treatment, and the concentration of RRx-001 that can keep the cell activity above 80% is below 2.5 μM.

[0135] To evaluate the combined treatment effect of G6PD inhibitor RRx-001 and doxorubicin on breast cancer MCF-7 cells, MCF-7 cells were randomly divided into five groups, one group as control, one group treated with 1.5 μM doxorubicin only, one group treated with 0.5 μM RRx-001, one group treated with 0.5 μM RRx-001 and 1.5 μM doxorubicin, and one group treated with 1.5 μM RRx-001 and 1.5 μM doxorubicin. After 48 h of treatment, the activity of the cells was detected using CCK-8 experiment, and the therapeutic effect of the drugs was evaluated by calculating the inhibition rate.

[0136] The experimental results are shown in Table 1. Figure 7 As shown in Table 1, the results show that the inhibition rate of the combination group is significantly higher than that of the single drug group, indicating that the combination of G6PD inhibitor RRx-001 and doxorubicin can significantly enhance the inhibition effect on MCF-7 cells, which indicates that the combination therapy can more effectively inhibit cell proliferation and improve the therapeutic effect of the drugs.

[0137] In the description of the present specification, the description of the terms "one embodiment", "some embodiments", "an example", "a specific example", or "some examples" and the like means that the specific features, structures, materials or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present application. In the present specification, the illustrative description of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any appropriate manner in any one or more embodiments or examples. In addition, different embodiments or examples described in the present specification and the features of different embodiments or examples can be combined and modified by those skilled in the art without contradiction.

[0138] Although the embodiments of the present application have been shown and described above, it is understood that the above-described embodiments are exemplary and should not be construed as limiting the present application, and those skilled in the art can make changes, modifications, replacements and variations to the above-described embodiments within the scope of the present application.

Claims

1. A combined drug for treating tumor drug resistance, characterized in that: include: Inhibitors of NADPH cycle-related genes and anti-tumor drugs.

2. The combined drug according to claim 1, characterized in that The dosage of the anti-tumor drug is an effective anti-tumor dose; Optionally, the dosage of the NADPH cycle-related gene inhibitor is a dosage required to maintain cell growth or activity at 30%-90% when acting alone, preferably, a dosage required to maintain cell growth or activity at 70%-80% when acting alone.

3. The combined drug according to claim 1, characterized in that The NADPH cycle-related genes include at least one of IDH2, IDH1, CBR1, G6PD, PGD, CBR3, FDXR, MTHFD1, MDH1, MDH2, DUS3L, MTHFD2, IDH3A, ALDH1B1, ALDH2, ALDH4A1, ALDH7A1, and PHGDH.

4. The combined drug according to claim 3, characterized in that The PHGDH inhibitor includes NCT-502, NCT-503, CBR-5884, PHGDH-IN-2, PHGDH-IN-3, PHGDH-IN-4, PHGDH-IN-5, BI-4916, BI-4924, or at least one of a pharmaceutically acceptable salt, ester or solvate thereof; Optionally, the IDH1 inhibitor includes at least one of AGI-5198, IDH-305, GSK864, or a pharmaceutically acceptable salt, ester or solvate thereof; Optionally, the IDH2 inhibitor includes at least one of AG-221, AG-881, AGI-12026, AGI-6780, IDH1 / 2-IN-1, GSK321, or a pharmaceutically acceptable salt, ester or solvate thereof; Optionally, the G6PD inhibitor includes at least one of RRx-001, G6PDi-1, polydatin, 6-aminonicotinamide, or a pharmaceutically acceptable salt, ester or solvate thereof; Optionally, the CBR1 inhibitor comprises at least one of quercetin 3-O-glucoside, rutin, or a pharmaceutically acceptable salt, ester or solvate thereof; Optionally, the MTHFD1 inhibitor comprises LY 345899, DS18561882, or at least one of a pharmaceutically acceptable salt, ester or solvate thereof; Optionally, the MDH1 inhibitor comprises MDH1-IN-1, MDH1-IN-2, or at least one of a pharmaceutically acceptable salt, ester or solvate thereof; Optionally, the MDH2 inhibitor comprises LW6, or a pharmaceutically acceptable salt, ester or solvate thereof; Optionally, the MTHFD2 inhibitor includes at least one of MTHFD2-IN1, MTHFD2-IN2, MTHFD2-IN3, MTHFD2-IN4, MTHFD2-IN5, DS44960156, TH9619, LY 345899, DS18561882, or a pharmaceutically acceptable salt, ester or solvate thereof; Optionally, the ALDH1B1 inhibitor comprises IGUANA-1, or a pharmaceutically acceptable salt, ester or solvate thereof; Optionally, the ALDH2 inhibitor includes at least one of Alda-1, Alda-2, daidzin, CVT-10216, 4-hydroxynonenal, or pharmaceutically acceptable salts, esters, or solvates thereof.

5. The combined drug according to claim 4, characterized in that The anti-tumor drug includes at least one of an alkylating agent, an antimetabolite, a plant alkaloid drug, an antibiotic drug, a small molecule kinase inhibitor, a monoclonal antibody drug, an immune checkpoint inhibitor, an epigenetic regulator, an antibody-drug conjugate, a metabolic targeted drug, and a PARP inhibitor; Optionally, the anti-tumor drug includes at least one of temozolomide, doxorubicin, cyclophosphamide, chlorambucil, carmustine, lomustine, cisplatin, oxaliplatin, carboplatin, 5-fluorouracil, cabinitabine, methotrexate, pemetrexed, gemcitabine, irinotecan, topotecan, etoposide, cytarabine, paclitaxel, docetaxel, bleomycin, actinomycin D, osimertinib, erlotinib, palbociclib, trastuzumab, rituximab, bevacizumab, vorinostat, azacitidine, olaparib, and niraparib.

6. The combined drug according to claim 5, characterized in that The inhibitor of NADPH cycle-related genes is RRx-001, the anti-tumor drug is temozolomide, and the molar ratio of RRx-001 to temozolomide is (0.5-2):(1000-2500); preferably, (1.3-2):(1050-2200); more preferably, (1.3-1.5):(1050-2200); Optionally, the inhibitor of NADPH cycle-related genes is AG-221, the anti-tumor drug is temozolomide, and the molar ratio of AG-221 to temozolomide is (1-4):(70-200); preferably, (2-4):(70-200); more preferably, (2.8-4):(140-200); Optionally, the inhibitor of NADPH cycle-related genes is AGI-5198, the anti-tumor drug is doxorubicin, and the molar ratio of AGI-5198 to doxorubicin is (50-500):(0.5-5); preferably, (100-200):(1-2); Optionally, the inhibitor of NADPH cycle-related genes is RRx-001, the anti-tumor drug is doxorubicin, and the molar ratio of RRx-001 to doxorubicin is (0.5-5):(0.5-5); preferably, (0.5-2):(0.5-2).

7. The combined drug according to claim 6, characterized in that The inhibitor of NADPH cycle-related genes and the anti-tumor drug are used simultaneously, separately or sequentially.

8. A pharmaceutical composition, characterized in that The invention comprises an inhibitor of NADPH cycle-related genes and an anti-tumor drug as active ingredients, and a pharmaceutically acceptable carrier.

9. Use of the combined drug according to any one of claims 1 to 7 or the pharmaceutical composition according to claim 8 in the preparation of a drug for treating tumor drug resistance.

10. The use according to claim 9, characterized in that The tumor includes at least one of brain glioma, glioblastoma, neuroblastoma, breast cancer, ovarian cancer, colorectal cancer, gastric cancer, lymphoma, sarcoma, testicular cancer, lung cancer, melanoma, and leukemia.

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