Methods of treating cancer using TH-302 monodrugs or in combination with PARP inhibitors

Abstract

The invention provides a method for treating cancer by single drug of TH-302 and analogues thereof or combination of the TH-302 and the analogues thereof with a PARP inhibitor, in particular to treatment of cancer patients sensitive to the PARP inhibitor by single drug of TH-302 and treatment of cancer patients by combination of TH-302 and the PARP inhibitor.

Description

Methods of treating cancer using TH-302 alone or in combination with a PARP inhibitor Technical Field

[0001] The present invention relates to methods for treating cancer, in particular methods for treating cancer patients who are sensitive to PARP inhibitors (PARPi). Background Art

[0002] The first human clinical trial of the PARPi drug Olaparib confirmed for the first time that PARPi can inhibit the growth of tumor cells carrying BRCA1 / 2 mutations. This is mainly based on the theory of synthetic lethality (Ashworth, A., & Lord, CJ (2018). Synthetic lethal therapies for cancer: what's next after PARP inhibitors?. Nature review. Clinical oncology, 15(9), 564–576. https: / / doi.org / 10.1038 / s41571-018-0055-6): PARP inhibitors can inhibit the DNA single-strand damage repair function of PARP, resulting in a large amount of single-strand DNA damage in the cell that cannot be repaired in time. Unrepaired single-strand DNA damage triggers replication fork disintegration and generates double-strand DNA damage. Double-strand DNA damage with strong cytotoxicity can be repaired in normal cells through the homologous recombination repair (HR) pathway mediated by proteins such as BRCA1 and BRCA2. However, in BRCA1 / 2-deficient tumor cells, double-strand DNA damage cannot be repaired, leading to the ultimate death of tumor cells. PARPi was originally developed for radiotherapy and chemotherapy sensitization, and preclinical studies also support the development of PARPi as a single drug for the treatment of BRCA1 / 2 gene-deficient cancers. Therefore, the initial target population for the validation of the PARPi-BRCA hypothesis was selected as carriers of BRCA1 / 2 germline mutations (gBRCA1 / 2). The initial study of PARPi in ovarian cancer enrolled patients who had previously received platinum chemotherapy, and the study found that platinum sensitivity was directly associated with PARPi response (platinum chemotherapy drugs are DNA-damaging agents that cause DNA cross-links, which can be partially repaired by the HR pathway; therefore, DNA repair-deficient tumors are expected to be sensitive to platinum chemotherapy). Two other PARPis have been approved in ovarian cancer: niraparib and rucaparib: the FDA and EMA have approved niraparib as a maintenance therapy (regardless of BRCA1 / 2 status); rucaparib has also been registered by the FDA and EMA as an optional treatment option for patients with BRCA1 / 2 mutation-related ovarian cancer who have previously received two lines of chemotherapy; and talazoparib has also been approved by the FDA for the treatment of BRCA-mutated / HER-2-negative metastatic breast cancer (Mateo, J., Lord, C.J., Serra, V., Tutt, A., J., Castroviejo-Bermejo, M., Cruz, C., Oakni n, A., Kaye, SB, & de Bono, JS (2019). A decade of clinical development of PA RP inhibitors in perspective. Annals of oncology: official journal of the European Society for Medical Oncology, 30(9),1437–1447. https: / / doi.org / 10.1093 / annonc / mdz192).

[0003] Summary of the Invention

[0004] TH-302 (Evofosfamide, CAS No. 918633-87-1) is a 2-nitroimidazole-triggered hypoxia-activated prodrug (HAP) of bromoisophosphamide. Under hypoxic conditions, the inactive TH-302 prodrug releases the highly toxic Br-IPM. TH-302 exhibits broad in vitro and in vivo biological activity, as well as specific hypoxia-selective activation, inducing H2AX phosphorylation and DNA cross-linking, leading to cell cycle arrest. Consequently, the compound is being developed as an anticancer drug by numerous pharmaceutical companies and research institutes.

[0005] A research article published by Meng FY et al. indicates that TH-302 has broad-spectrum activity against various tumors and exhibits excellent hypoxia-selective activity enhancement. The study demonstrated that the in vitro cytotoxicity of TH-302 in 32 human cancer cell lines under hypoxia was significantly stronger than that under normoxic conditions, indicating that the compound has selective cytotoxicity against cancer cells in hypoxic environments. Human cells overexpressing the single-electron reductase (POR) were used to confirm the single-electron reductase-dependent activity enhancement mechanism of TH-302 under hypoxia, as shown in the following reaction formula 1:

[0006] Cytochrome P450 oxidoreductase reduces the prodrug TH-302 to produce an intermediate free radical anion. This unstable free radical anion then decomposes into the cytotoxic Br-IPM. The key step in this process is the one-electron reduction process. Studies have shown that the presence of oxygen reverses this one-electron reduction process. Therefore, TH-302 can only be reduced to produce the more cytotoxic Br-IPM in an oxygen-deficient environment. Furthermore, the in vitro cytotoxicity of TH-302 was tested using cell lines based on Chinese hamster ovary cells with DNA repair mutations, including those deficient in base excision, nucleotide excision, non-homologous end-joining repair, or homologous end-joining repair (i.e., homology-dependent repair). The study found that cell lines deficient in homologous end-joining repair alone or in combination with nucleotide excision repair showed significantly increased hypoxic sensitivity to TH-302. However, cell lines deficient in base excision, nucleotide excision, or non-homologous end-joining repair alone had no effect on TH-302 sensitivity. Consistent with this finding, enhanced sensitivity to TH-302 was also observed in in vitro experiments of cells lacking BRCA1, BRCA2, and FANCA DNA double-strand repair; and in clinical trials, it was also observed that TH-302 had a better therapeutic effect on patients with BRCA gene mutations (Meng F, Evans JW, Bhupathi D, et al. Molecular and cellular pharmacology of the hypoxia-activated prodrug TH-302. [J]. Molecular Cancer Therapeutics, 2012, 11(3): 740; Conroy, M., Borad, MJ, & Bryce, AH (2017). Hypoxia-Activated Alkylating Agents in BRCA1-Mutant Ovarian Serous Carcinoma. Cureus, 9(7), e1517. https: / / doi.org / 10.7759 / cureus.1517; WO2012135757A2, Methods for treating cancer; WO2015013448A1, Treatment of pancreatic cancer with a combination of a hypoxia-acti vated prodrug and a taxane; WO2020007106A1, Anticancer medical use of efsomide).

[0007] These studies on the mechanism of action of TH-302, particularly its exceptional sensitivity to BRCA mutations, suggest that TH-302 may have particularly significant therapeutic effects in certain cancer patients. The inventors' experimental results further demonstrate that TH-302 is more effective than PARP inhibitors in treating cancer, even in patients with BRCA pathogenic mutations for which PARP inhibitors are highly responsive. To this end, this application provides the following cancer treatment methods.

[0008] A method of treating cancer or tumor patients using a drug containing a hypoxia-activated compound of formula (1) alone or in combination with a PARP inhibitor:

[0009] Wherein, R is independently selected from H, -CH3, -CH2CH3, -CF3, and X is independently selected from leaving functional groups such as Cl, Br, MsO, and TsO.

[0010] The drug described herein refers to a medicine or preparation, wherein the prepared medicine comprises the hypoxia-activated compound of formula (1) or its salt or solvate in a specific dosage range as the active ingredient, and / or the prepared medicine is in a specific dosage form and is administered by a specific administration method.

[0011] The prepared medicine, drug, preparation may also include pharmaceutically acceptable adjuvants or excipients. The medicine may be in any dosage form for clinical use, such as tablets, suppositories, dispersible tablets, enteric-coated tablets, chewable tablets, orally disintegrating tablets, capsules, sugar-coated tablets, granules, dry powders, oral solutions, small needles for injection, freeze-dried powder for injection, or large infusions. Depending on the specific dosage form and mode of administration, the pharmaceutically acceptable adjuvants or excipients in the medicine may include one or more of the following: diluents, solubilizers, disintegrants, suspending agents, lubricants, adhesives, fillers, flavorings, sweeteners, antioxidants, surfactants, preservatives, wrappers, and pigments.

[0012] With TH-302 or its similar compounds Related preparations include oral preparations, lyophilized preparations and concentrated injections, and the related prescriptions, preparation methods, clinical compatibility and administration methods are described and disclosed in detail in Threshold's relevant patents: WO2010048330A1, WO2012142520A2 and WO2008083101A1. The present invention hereby introduces the full text of the above application texts.

[0013] TH-302 or its similar compounds It is a DNA alkylating anticancer drug with a wide range of cancer therapeutic potential. These related cancer indication experiments and clinical trials are disclosed in the patent applications of Threshold and other pharmaceutical companies (such as WO2016011195A2, WO2004087075A1, WO2007002931A1, WO2008151253A2, WO2009018163A1, WO2009033165A2, WO2010048330A2, WO2012142520A1, WO2008083101A2, WO2020007106A1, WO2020118 251A1, WO2014169035A1, WO2013116385A1, WO2019173799A2, WO2016081547A1, WO2014062856A1, WO2015069489A1, WO2012006032A2, WO201802 6606A2, WO2010048330A2, WO2015171647A1, WO2013096687A1, WO2013126539A2, WO2013096684A2, WO2012009288A2, WO2012145684A2, WO201601 4390A2, WO2019055786A2, WO2012135757A2, WO2015013448A2, WO2016011328A2, WO2013177633A2, WO2016011195A2, WO2015051921A2) and FDA-registered clinical trials (NCT02402062, NCT02020226, NCT02076230, NCT01381822, NCT02093962, NCT01440088, NCT02255110, NCT02342379, NCT01864538, NCT01 149915, NCT02433639, NCT00743379, NCT01485042, NCT01721941, NCT02047500, NCT00742963, NCT01497444, NCT00495144, NCT01746979, NCT01144455, NCT01403610, NCT01522872, NCT01833546, NCT02598687, NCT03098160, NCT02496832, NCT02712567). The present invention hereby introduces all the above-mentioned related application texts and clinical trial information.

[0014] "Cancer" refers to leukemias, lymphomas, carcinomas, and other malignancies (including solid tumors) with potentially unrestricted growth that can spread locally by invasion and systemically by metastasis.

[0015] Here are TH-302 or its similar compounds Examples of cancers that can be treated include, but are not limited to, cancers of the adrenal gland, bone, brain, breast, bronchi, colon and / or rectum, gallbladder, head and neck, kidney, larynx, liver, lung, nervous tissue, pancreas, prostate, parathyroid gland, skin, stomach, and thyroid gland. Some other examples of cancer include acute and chronic lymphocytic and granulocytic tumors, adenocarcinomas, adenomas, basal cell carcinomas, dysplastic epithelial cells and carcinoma in situ of the cervix, Ewing's sarcoma, epidermoid carcinoma, giant cell tumors, glioblastoma multiforme, hair cell tumors, intestinal ganglioneuromas, proliferative corneal nerve tumors, pancreatic islet cell carcinoma, Kaposi's sarcoma, leiomyoma, leiomyoma, leukemia, lymphoma, malignant carcinoid tumors, malignant melanoma, malignant hypercalcemia, Marfanoid body-type tumors, medullary epithelial carcinoma, metastatic skin cancer, mucosal neuroma, myeloma, mycosis fungoides, neuroblastoma, osteosarcoma, osteogenic and other sarcomas, ovarian tumors, Pheochromocytoma, polycythemia vera, primary brain tumors, small cell lung cancer, squamous cell carcinoma of both the ulcerative and papillary types, hyperplasia, seminoma, soft tissue sarcoma, retinoblastoma, rhabdomyosarcoma, renal cell tumor, localized skin lesions, reticulum cell sarcoma, and Wilm's tumor.

[0016] PARP is an enzyme, known as poly (ADP-ribose) polymerase (PARP). PARP is a single-strand DNA repair enzyme that plays a key role in the DNA repair pathway. When a single strand of DNA is damaged and broken, PARP is activated. As a molecular sensor of DNA damage, it recognizes and binds to the DNA break site, activating and catalyzing the poly (ADP-ribosyl)ation of the receptor protein, thus participating in the DNA single-strand repair process.

[0017] PARP inhibitors inhibit the work of PARP enzymes, preventing these PARP enzymes, which are equivalent to repair workers, from working normally. Single-strand damage to DNA cannot be repaired and cells will die.

[0018] Because cells have more than just PARP as their repairman, even if PARP fails and DNA damage is carried to the next stage of the process, another repairman, such as double-strand repair, is waiting in the wings to repair the DNA. The protein produced by the BRCA gene is a key member of this second repairman. Normal cells possess this double-safety mechanism; even if PARP inhibitors disrupt one layer of damage, the other remains functional, preventing cell death.

[0019] However, in ovarian or breast cancer cells with BRCA gene mutations, the BRCA repairman no longer works properly. Of course, because the PARP team continues to work, the cancer cells will not die.

[0020] If PARP inhibitors are used to disable PARP, the DNA of cancer cells cannot be repaired. In this way, PARP inhibitors can kill only cancer cells without killing normal cells.

[0021] PARP inhibitors + BRCA gene mutations together are what we call "synthetic lethality". Simply put, synthetic lethality means that when two different genes (BRCA) or proteins (PRAP) lose their function at the same time, it will cause cell death, while if only one of these two genes / proteins loses its function, it will not cause cell death.

[0022] PARP inhibitors are compounds that have an inhibitory effect on PARP enzymes, that is, all substances that can inhibit the activity of PARP enzymes are PARP inhibitors.

[0023] The five drugs selected from the marketed drugs are Olaparib, Rucaparib, Niraparib, Talazoparib, Fluzoparib and Pamiparib which have entered the clinical phase III. Obviously, the PARP inhibitors here actually refer to drugs containing the active ingredients of PARP inhibitors.

[0024] Talazoparib is indicated for adults with deleterious or suspected deleterious germline BRCA-mutated (gBRCAm) HER2-negative locally advanced or metastatic breast cancer. Talazoparib is available in 0.25 mg / 1 mg talazoparib tosylate capsules, taken orally once daily. Treatment interruption or dose reduction should be considered in the event of adverse reactions:

[0025] At the first occurrence of an adverse reaction, the oral dose was reduced to 0.75 mg (three 0.25 mg capsules) once daily;

[0026] If the adverse reaction occurs a second time, the oral dose is reduced to 0.5 mg (two 0.25 mg capsules) once daily;

[0027] The third time an adverse reaction occurred, the oral dose was reduced to 0.25 mg (one 0.25 mg capsule) once daily.

[0028] Niraparib is used for the maintenance treatment of adult patients with platinum-sensitive recurrent epithelial ovarian, fallopian tube, or primary peritoneal cancer who have achieved complete or partial remission after platinum-containing chemotherapy. The commercial dosage form is 100 mg of niraparib tosylate capsules, taken orally once a day at 300 mg until disease progression or intolerable adverse reactions occur. In the event of adverse reactions, treatment interruption or dose reduction should be considered:

[0029] Dose reduction was first from 3 capsules (300 mg) per day to 2 capsules (200 mg) per day;

[0030] If further dose reduction is required, the dose can be reduced a second time from 2 capsules (200 mg) per day to 1 capsule (100 mg) per day;

[0031] If suspending the drug and reducing the dose cannot control the adverse reaction, discontinuation of the drug is recommended.

[0032] Rucaparib is used for women with advanced ovarian cancer whose tumors carry a specific gene mutation (harmful BRCA) and who have been treated with two or more chemotherapy drugs. It is available in three tablet forms: 200 mg, 250 mg, and 300 mg. The recommended dose is 600 mg, taken orally twice daily, with or without food. Continue treatment until disease progression or unacceptable toxicity. For adverse reactions, consider interrupting treatment or reducing the dose.

[0033] Olaparib is used for maintenance treatment of adult patients with advanced epithelial ovarian, fallopian tube, or primary peritoneal cancer who have germline or somatic BRCA mutations (gBRCAm or sBRCAm) after achieving complete remission or partial remission with platinum-containing chemotherapy; and for maintenance treatment of adult patients with platinum-sensitive recurrent epithelial ovarian, fallopian tube, or primary peritoneal cancer after achieving complete remission or partial remission with platinum-containing chemotherapy. The marketed dosage form is tablets: 150 mg and 100 mg, with two specifications. The recommended dose is 300 mg (2 150 mg tablets) twice a day, equivalent to a total daily dose of 600 mg. The 100 mg tablet is used for dose reduction:

[0034] To manage adverse events, such as nausea, vomiting, diarrhea, and anemia, interruption of treatment or dose reduction may be considered;

[0035] If dose reduction is necessary, the recommended dose is reduced to 250 mg (one 150 mg tablet, one 100 mg tablet) taken twice daily (equivalent to a total daily dose of 500 mg);

[0036] If further dose reduction is necessary, the recommended dose is reduced to 200 mg (two 100 mg tablets) taken twice daily (equivalent to a total daily dose of 400 mg).

[0037] Fluzoparib is used to treat patients with platinum-sensitive recurrent ovarian, fallopian tube, or primary peritoneal cancer who have undergone two or more lines of chemotherapy and harbor germline BRCA mutations (gBRCAm). The commercially available dosage form is a 50mg capsule.

[0038] For other PARPi candidate drugs under development and entering clinical practice, please refer to the web link https: / / www.selleckchem.com / PARP.html and related academic review literature.

[0039] TH-302 or its similar compounds The recommended dosage for treating cancer can be found in patent applications filed by Threshold and other pharmaceutical companies (e.g., WO2016011195A2, WO2004087075A1, WO2007002931A1, WO2008151253A2, WO2009018163A1, WO2009033165A2, WO2010048330A2, WO2012142520A1, WO2008083101A2, WO2020007106A1, WO2020118 251A1, WO2014169035A1, WO2013116385A1, WO2019173799A2, WO2016081547A1, WO2014062856A1, WO2015069489A1, WO20 12006032A2, WO2018026606A2, WO2010048330A2, WO2015171647A1, WO2013096687A1, WO2013126539A2, WO2013096684A2, WO2012009288A2, WO2012145684A2, WO2016014390A2, WO2019055786A2, WO2012135757A2, WO2015013448A2, WO2016011328A2, WO2013177633A2, WO2016011195A2, WO2015051921A2) and FDA-registered clinical trials (NCT02402062, NCT02020226, NCT02076230, NCT01381822, NCT02093962, NCT01440088, NCT0225511) 0, NCT02342379, NCT01864538, NCT01149915, NCT02433639, NCT00743379, NCT01485042, NCT01721941, NCT02047500, NCT00742963, NCT01497444, NCT00495144, NCT01746979, NCT01144455, NCT01403610, NCT01522872, NCT01833546, NCT02598687, NCT03098160, NCT02496832, NCT02712567).

[0040] 120 mg / m 2 Up to 460 mg / m 2 The daily dose is administered intravenously;

[0041] 480 mg / m 2 Up to approximately 670 mg / m 2 Or for example 575 mg / m 2 The weekly dose is administered intravenously.

[0042] TH-302 (concentrate for administration solution) used in clinical trials is a sterile liquid formulation of TH-302. TH-302 is formulated with 70% anhydrous ethanol, 25% dimethylacetamide, and 5% polysorbate 80. It is provided by the sponsor in 10 mL glass vials with rubber stoppers and flip-off closures. The TH-302 drug product is a clear, colorless to pale yellow solution that is essentially free of visible particulates. For a nominal total amount of 650 mg of TH-302, each single-use vial contains a nominal fill volume of 6.5 mL of TH-302 drug product (equivalent to 100 mg / mL) and is clearly labeled, disclosing the batch number, route of administration, required storage conditions, the sponsor's name, and appropriate precautionary labeling as required by applicable regulations. Dilution is required according to the pharmacy manual prior to administration.

[0043] Prior to administration, dilute to a total volume of 500 mL (1000 mL for a total dose of ≥1000 mg) with a commercially available 5% dextrose aqueous solution to achieve the desired final concentration. Each dose of TH-302 was prepared with a 5% dextrose aqueous solution that was free of di(2-ethylhexyl) phthalate (DEHP) and administered intravenously using a DEHP-free intravenous infusion device.

[0044] Of course, you can also use the freeze-dried preparation developed by Threshold:

[0045] A solution (20 mL) of TH-302 (100 mg) and sucrose (1 g) was added to lyophilization vials and lyophilized to produce lyophilized unit dosage forms of TH-302 with a drug loading of less than 5 mg / cm 3 For human administration, the unit dosage form is dissolved in 5% dextrose injection and an appropriate amount of this solution is administered to the patient;

[0046] The subsequent Phase I clinical trial dosing regimen for TH-302 in human patients used a lyophilized formulation. The lyophilized formulation of TH-302 for injection was prepared in a 100 mL glass vial with a drug loading of 100 mg / 100 ml and stored under controlled conditions at 2-8°C. When used, the formulation was injected into 250 mL of 5% glucose injection in the lyophilized formulation bottle and administered intravenously within 30 minutes via an infusion pump.

[0047] Monotherapy refers to single-drug therapy. Combination therapy refers to combined drug therapy. Monotherapy refers to the use of only one anticancer drug in a single course of treatment. Combination therapy refers to the use of two or more anticancer drugs simultaneously or sequentially in a single course of treatment.

[0048] Generally speaking, combination therapy requires exploring different dosages and dosing cycles based on the characteristics of the disease and the types of combined drugs. Only based on the above conditions can the combination drug treatment plan explored achieve better therapeutic effects than single drug treatment.

[0049] The drug dosage and administration cycle of monotherapy and combination therapy need to be explored through clinical trials with reference to the dosage and administration regimen of the above-mentioned TH-302 and its similar compounds and PARPi.

[0050] Furthermore, the patient's DNA repair enzymes are impaired.

[0051] According to relevant research literature, the damaged DNA repair enzymes are selected from:

[0052] Damage to homologous recombination DNA repair enzymes,

[0053] Damage to nucleotide excision repair enzymes,

[0054] Impaired nonhomologous end joining enzymes

[0055] Damage to base excision repair enzymes,

[0056] Damage to mismatch repair enzymes

[0057] One or more of the Fanconi's anemia pathway repair enzymes are impaired.

[0058] Preferably, any one or more of the following are impaired homologous recombination DNA repair enzymes, impaired nucleotide excision repair enzymes, and impaired base excision repair enzymes. More preferably, only the homologous recombination DNA repair enzyme is impaired, or both the homologous recombination DNA repair enzyme and the nucleotide excision repair enzyme are impaired.

[0059] Furthermore, the patient's tumor or cancer tissue is detected to have any one or two pathogenic gene mutations in the genes corresponding to BRCA1 and BRCA2; or the patient is detected to have any one or two pathogenic gene mutations in the genes corresponding to BRCA1 and BRCA2.

[0060] Mutations in either or both of the BRCA1 and BRCA2 genes can be detected using commercially available (companion) diagnostic kits:

[0061] The BRACAnalysisCDx Olaparib companion test kit, developed by Myriad Pharmaceuticals in the United States, is a genetic test used to detect BRCA gene mutations in blood samples from patients with ovarian cancer;

[0062] The BRCA1 / 2 gene mutation detection kit (combined probe-anchored polymerase sequencing method), developed by BGI Biotech (Wuhan) Co., Ltd., is used to qualitatively detect germline mutations in the exon regions and adjacent intron regions of the BRCA1 / 2 gene in patients clinically diagnosed with ovarian and breast cancer;

[0063] The human BRCA1 and BRCA2 gene mutation detection kit (reversible terminal termination sequencing method) was developed by Xiamen Aide Biotechnology Co., Ltd. and can be used for relevant medication guidance of the PARP inhibitor Olaparib tablets.

[0064] BRCA1 and BRCA2 mutations include germline mutations (gBRCAm) and somatic mutations (sBRCAm) of BRCA1 and BRCA2 mutations.

[0065] The hypoxia-activated compound of formula (1) is selected from the compounds of the following structures:

[0066] TH-302, and In particular, TH-302 is preferred.

[0067] Furthermore, the cancer or tumor is selected from ovarian cancer, breast cancer, pancreatic cancer, fallopian tube cancer, primary peritoneal cancer, gastric cancer, prostate cancer, non-small cell lung cancer, small cell lung cancer, liver cancer, colon cancer, rectal cancer, etc.

[0068] The TMB (tumor mutation burden) level of the BRCA pathogenic gene mutation is medium.

[0069] Because TMB (tumor mutation burden) varies among different tumor types, it's generally considered high if it exceeds 20 mutations / Mb (Mb stands for bases per million), low if it's below 10 mutations / Mb, and moderate if it's in between. At the 2017 World Lung Cancer Conference, Bristol-Myers Squibb announced the results of the CheckMate-032 clinical trial, a Phase II study that enrolled 401 patients with advanced lung cancer who had failed first-line therapy. The patients were treated with a PD-1 inhibitor alone or in combination with ipilimumab. Patients were divided into three groups based on TMB levels: high, intermediate, and low. Among those receiving the combination therapy, the response rate was 62%, 20%, and 23%, respectively, with the response rate three times higher in those with high TMB. The median overall survival was 22.0 months, 3.6 months, and 3.4 months, respectively—a six-fold difference! This study demonstrated that different TMB levels have a significant impact on the efficacy of different cancer treatment drugs.

[0070] The cancer or tumor patients are sensitive or resistant to PARP inhibitors.

[0071] Sensitivity and drug resistance are qualitative judgments that evaluate whether a patient is resistant to a drug, and are commonly seen in drug sensitivity tests of microbial pathogens and antibacterial (bacterial and fungal) drugs.

[0072] Sensitivity and drug resistance are two different results that appear during clinical drug sensitivity tests. Sensitivity means that during the drug sensitivity test, it was found that the pathogen was very sensitive to the drug, which means that the drug can kill all pathogens; while drug resistance means that during the experiment, it was found that the drug could not kill the pathogen, which means that the drug is ineffective. In addition, when this result is obtained clinically, it is very meaningful for treatment, and a more sensitive drug can be selected to kill the pathogen. At the same time, there is also a result called moderate sensitivity, which means that it is moderately sensitive and can easily develop drug resistance but may also be useful, but the efficacy will not be very good and will not be as effective as sensitive treatment.

[0073] In chemotherapy for cancer treatment (compared to surgery and radiotherapy, chemotherapy here is broadly defined, referring to the use of drugs to treat cancer), according to clinical treatment guidelines, a series of chemotherapy drugs are usually used in sequence: when the first-administered drug has no therapeutic effect on the patient after a period of administration or the therapeutic effect is very weak, it is considered that the cancer patient has developed resistance or tolerance to this drug, and other drugs (second-line drugs) are needed. Correspondingly, second-line drugs may also develop resistance or resistance after a period of administration, and third-line drugs are needed. Sensitivity and resistance are diagnosed and judged by medical professionals or clinical physicians.

[0074] Obviously, the specific meaning of cancer and tumor patients being sensitive to PARP inhibitors is that if the patient is diagnosed with a BRCA pathogenic mutation and responds to PARP inhibitors or is judged to be effective after diagnosis and treatment by a doctor, in this case, TH-302 alone or a PARP inhibitor combined with TH-302 will have a better therapeutic effect.

[0075] That is to say, the administration of TH-302 according to the present invention will replace PAPR inhibitors, and TH-302 can replace all the indications of PAPR inhibitors: the indications of marketed PAPR inhibitors include platinum-sensitive recurrent epithelial ovarian cancer, fallopian tube cancer or primary peritoneal cancer; advanced epithelial ovarian cancer, fallopian tube cancer or primary peritoneal cancer carrying germline or somatic BRCA mutations. The corresponding TH-302 may be effective for these indications, and the efficacy is better than PAPR inhibitors.

[0076] The present invention also provides a method of treating, comprising the following steps:

[0077] Detect BRCA1 and BRCA2 gene mutations in cancer and tumor patients;

[0078] If the patient has a BRCA1 or BRCA2 gene mutation, the patient is treated with a drug containing a hypoxia-activated compound of formula (1) alone or in combination with a PARP inhibitor:

[0079] Wherein, R is independently selected from H, -CH3, -CH2CH3, -CF3, and X is independently selected from leaving functional groups such as Cl, Br, MsO, and TsO.

[0080] In addition to containing the hypoxia-activated compound of formula (1), the above-mentioned drug should also be added with pharmaceutically acceptable excipients or vehicles according to the specific characteristics of the drug, medicine, or preparation. The drug can be in any dosage form for clinical administration, such as tablets, suppositories, dispersible tablets, enteric-coated tablets, chewable tablets, orally disintegrating tablets, capsules, sugar-coated tablets, granules, dry powders, oral solutions, small needles for injection, freeze-dried powder for injection, or large infusions. Depending on the specific dosage form and administration method, the pharmaceutically acceptable excipients or vehicles in the drug can include one or more of the following: diluents, solubilizers, disintegrants, suspending agents, lubricants, adhesives, fillers, flavoring agents, sweeteners, antioxidants, surfactants, preservatives, encapsulating agents, and pigments.

[0081] When used, the above drugs can be used alone or in combination with PARPi drugs for treatment.

[0082] A preparation unit package, comprising an independent packaging container containing a pharmaceutical preparation, an outer packaging component for accommodating the independent packaging container, and a drug package insert, wherein the pharmaceutical preparation contains a hypoxia-activated compound of formula (I), and the drug package insert states that the drug is used in combination with a PARP inhibitor to treat cancer and tumor patients:

[0083] Wherein, R is independently selected from H, -CH3, -CH2CH3, -CF3, and X is independently selected from leaving functional groups such as Cl, Br, MsO, and TsO.

[0084] Preparation unit packaging is a concept in the process of drug production and sales. It refers to the process in which drugs (medicaments) are loaded into independent inner packaging containers and labels are affixed or printed on the outer wall of the inner packaging containers. Finally, the independent packaging container with the label and the medicine inside is placed in the outer packaging component (usually a paper packaging box that is directly seen by consumers), and the drug instructions (label) containing the most critical information about the drug are also placed in the outer packaging component together with the independent packaging container.

[0085] Usually the legal drug label contains the following legal contents:

[0086] For the United States, the drug label must record

[0087] Boxed Warning (some drugs have serious side effects)

[0088] 1. Indications and Usage

[0089] 2. Dosage and Administration

[0090] 3. Dosage Forms and Strengths

[0091] 4. Contraindications

[0092] 5.Warnings and Precautions

[0093] 6.Adverse Reactions

[0094] 7.Drug Interactions

[0095] 8. Use in Specific Populations

[0096] 9.Drug Abuse and Dependence (not available for some drugs)

[0097] 10. Overdosage

[0098] 11. Description (including chemical name, molecular formula, molecular weight, structural formula, physical and chemical properties)

[0099] 12.Clinical Pharmacology

[0100] 13. Nonclinical Toxicology

[0101] 14.Clinical Studies

[0102] 15. References (some drugs do not have)

[0103] 16.How Supplied / Storage and Handling

[0104] 17. Patient Counseling Information

[0105] In China, drug inserts must record the drug name, ingredients (active ingredients, excipients), properties, specifications, indications, usage and dosage, contraindications, warnings and precautions, adverse reactions, drug interactions, use in pregnant and lactating women, use in children, use in the elderly, drug abuse and drug dependence, drug overdose, clinical pharmacology (pharmacology, pharmacokinetics, pharmacogenetics), pharmacology and toxicology (pharmacological effects, toxicological studies), clinical trials, storage, packaging, expiration date, implementation standards, approval number, drug marketing authorization holder (name, registered address), manufacturer (company name, production address), etc.

[0106] It can be said that the drug instructions in the final sales unit packaging are the key content for the use of the drug: without the drug instructions, the drug will not be able to be used. Therefore, the drug instructions are indispensable for the final function of the drug and are also an inevitable link in the drug research and development and manufacturing process. In fact, it is impossible for drug research and development and manufacturing related entities (drug research and development manufacturers, drug sales agencies, medical institutions) to directly produce drugs for specific single drug / combination indications without research activities such as research and development and clinical trials, and it is even more impossible to sell, distribute and administer to patients. Therefore, the content of the drug instructions is very important and is a key and indispensable link in the drug manufacturing process (drugs that are finally administered must have drug instructions).

[0107] The drug is used in combination with PARP inhibitors to treat cancer and tumor patients, which falls under the drug's Indications and Usage (United States) and Indications (China).

[0108] Of course, the preparation unit packaging of the above-mentioned drugs can also be in the form of pharmaceutical use:

[0109] The pharmaceutical use of the hypoxia-activated compound of formula (I) in the preparation of a dosage unit package is achieved by the following operations:

[0110] Packing the drug preparation into individual packaging containers;

[0111] The individual packaging container containing the medicine is placed into the outer packaging component;

[0112] Place the drug leaflet into the outer packaging component,

[0113] It is characterized in that the pharmaceutical preparation contains the hypoxia-activated compound of formula (I), and the drug instructions record that the drug is used in combination with a PARP inhibitor to treat cancer and tumor patients:

[0114] Wherein, R is independently selected from H, -CH3, -CH2CH3, -CF3, and X is independently selected from leaving functional groups such as Cl, Br, MsO, and TsO.

[0115] Preferably, the patient's DNA repair enzyme is impaired,

[0116] Preferably, the patient's tumor or cancer tissue is detected to have any one or two pathogenic gene mutations in the genes corresponding to BRCA1 and BRCA2; or the patient is detected to have any one or two pathogenic gene mutations in the genes corresponding to BRCA1 and BRCA2,

[0117] Preferably, BRCA1 and BRCA2 mutations include germline mutations (gBRCAm) and somatic mutations (sBRCAm) of BRCA1 and BRCA2 mutations.

[0118] Preferably, the hypoxia-activated compound of formula (I) is selected from the compounds of the following structures:

[0119] Preferably, the PARP inhibitor is selected from Olaparib, Rucaparib, Niraparib, Talazoparib, Fluzoparib, Pamiparib,

[0120] Preferably, the cancer or tumor is selected from ovarian cancer, breast cancer, pancreatic cancer, fallopian tube cancer, primary peritoneal cancer, gastric cancer, prostate cancer, non-small cell lung cancer, small cell lung cancer, liver cancer, colon cancer, and rectal cancer.

[0121] Preferably, the cancer or tumor patient is sensitive or resistant to PARP inhibitors. BRIEF DESCRIPTION OF THE DRAWINGS

[0122] FIG1 is a growth curve of the tumor volume of mice in each group in the in vivo efficacy experiment of TH-302 and Olaparib in the Olaparib-sensitive OV5304 PDX model;

[0123] FIG2 is a curve showing the percentage change in body weight of mice in each group in the in vivo efficacy experiment of TH-302 and Olaparib in the Olaparib-sensitive OV5304 PDX model;

[0124] FIG3 is a graph showing the growth curve of tumor volume in each group of mice treated with TH-302 and Olaparib in the Capan-1 CDX pancreatic cancer model;

[0125] Figure 4 is a curve chart of the percentage change in body weight of mice in each group in the pancreatic cancer Capan-1 CDX model treated with TH-302 and Olaparib. DETAILED DESCRIPTION

[0126] The present invention is described below with reference to specific examples. Those skilled in the art will appreciate that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention in any way.

[0127] The experimental methods in the following examples are conventional methods unless otherwise specified. The medicinal materials, reagents, etc. used are all commercially available products unless otherwise specified.

[0128] "Patient" and "subject" are used interchangeably to refer to a mammal in need of cancer treatment. Typically, the patient is a human. Typically, the patient is a human diagnosed with cancer. In certain embodiments, a "patient" or "subject" may refer to a non-human mammal, such as a non-human primate, dog, cat, rabbit, pig, mouse, or rat, used to screen, characterize, and evaluate drugs and therapies.

[0129] "Prodrug" refers to a compound that, after administration or administration, is metabolized or otherwise converted to a biologically active or more active compound (or drug) with respect to at least one property. A prodrug is chemically modified relative to a drug in a manner that renders it less active or inactive relative to the drug, but the chemical modification allows for the production of the corresponding drug by metabolism or other biological processes following administration of the prodrug. A prodrug may have altered metabolic stability or delivery characteristics, fewer side effects or lower toxicity, or an improved flavor relative to the active drug. Prodrugs may be synthesized using reactants other than the corresponding drug.

[0130] "Treatment" or "treating a patient" refers to administering, using or applying to a patient a therapeutically effective amount of a drug related to the present invention.

[0131] "Administering" or "applying" a drug to a patient refers to direct administration or administration (which may be administered or administered to a patient by a medical professional or may be self-administered or administered) and / or indirect administration or administration, which may be the act of prescribing a drug. For example, a physician who instructs a patient to self-administer or administer a drug and / or provides a prescription for a drug to a patient is administering or administering a drug to a patient.

[0132] A "therapeutically effective amount" of a drug refers to an amount of the drug that, when administered or used to a patient suffering from cancer, will have the desired therapeutic effect (e.g., alleviation, amelioration, remission, or elimination of the clinical manifestations of one or more cancers in the patient). The therapeutic effect does not necessarily occur by administering or applying a single dose and may only occur after administering or applying a series of doses. Thus, a therapeutically effective amount can be administered or applied in one or more doses.

[0133] "Treatment" of a condition or patient refers to taking steps to obtain beneficial or desired results (including clinical results). For the purposes of this invention, beneficial or desired clinical results include, but are not limited to, alleviation or amelioration of one or more symptoms of cancer; reduction in the extent of the disease; delay or slowing of disease progression; improvement, remission, or stabilization of the disease state; or other beneficial results. In some instances, treatment of cancer may result in a partial response or stabilization of the disease.

[0134] "Tumor cell" refers to a tumor cell of any appropriate species (eg, mammalian, such as murine, canine, feline, equine, or human).

[0135] The above description of the specific embodiments of the present invention does not limit the present invention. Those skilled in the art can make various changes or modifications based on the present invention. As long as they do not depart from the spirit of the present invention, they should fall within the scope of the claims attached to the present invention.

[0136] The following provides specific experiments of the present invention.

[0137] 1. Efficacy and Safety Evaluation of TH-302 in the Olaparib-Sensitive OV5304 PDX Model (Experiment E4354-B2136)

[0138] To evaluate the in vivo efficacy of TH-302 in an Olaparib-sensitive model, we selected the OV5304 PDX model of ovarian cancer with a BRCA pathogenic mutation and Olaparib sensitivity. This model carries a BRCA1 pathogenic mutation site p.F580Kfs.

[0139] BALB / c nude female mice were inoculated subcutaneously The OV5304 tumor was used to establish a subcutaneous transplant model of human ovarian cancer. The study was divided into a glucose injection control group, a test drug olaparib 50 mg / kg monotherapy group, a test drug TH-302 20 mg / kg, 40 mg / kg, and 80 mg / kg monotherapy group, and a combination of olaparib 50 mg / kg and TH-302 40 mg / kg group. The glucose injection control group, the test drug TH-302 20 mg / kg, 40 mg / kg, and 80 mg / kg monotherapy groups, and the olaparib 50 mg / kg and TH-302 40 mg / kg combination group were administered via tail vein injection once weekly for three consecutive weeks. The test drug, olaparib 50 mg / kg alone, and the olaparib 50 mg / kg and TH-302 40 mg / kg combination group, were administered orally by gavage once daily for 30 consecutive days. The specific dosing schedule and TGI are shown in Table 1.

[0140] Table 1. Efficacy analysis data of each group in the ovarian cancer OV5304 model Note: 1. Data are expressed as "mean ± standard error", i.e. 2. T / C% = T RTV / C RTV ×100% or T / C%=T TV / C TV ×100%(T RTV : Average RTV of treatment group; CRTV : Average RTV of vehicle control group; RTV = V t / V0, V0 is the tumor volume of the animal when grouped, V t is the tumor volume of the animal after treatment, the same below); 3. TGI% = (1-T / C) x 100% (T and C are the average relative tumor volumes (RTV) of the treatment group and the control group, respectively, at a specific time point).

[0141] The corresponding curves of tumor volume and percentage of mouse body weight changing with time are shown in Figures 1 and 2.

[0142] Efficacy: The results of the efficacy experiment showed that in patients with BRCA1 pathogenic mutations In the ovarian cancer OV5304 model, the test drug Olaparib (50 mg / kg) monotherapy group had a significant tumor inhibition effect on day 30 after the first dose, with a relative tumor inhibition rate (TGI) of 96.16%, which was statistically significantly different from the control group (p<0.001). The test drug TH-302 20 mg / kg treatment group, TH-302 40 mg / kg, and TH-302 80 mg / kg monotherapy groups also showed a significant tumor inhibition effect on day 30 after the first dose, with relative tumor inhibition rates (TGI) of 82.81%, 97.98%, and 100%, respectively, which were statistically significantly different from the control group (p<0.01, p<0.001, and <0.001, respectively). TH-302 monotherapy showed a dose-dependent effect in this model.

[0143] We used an intermediate TH-302 monotherapy dose of 40 mg / kg in combination with olaparib at a 50 mg / mg dose to test whether the two monotherapy agents had a synergistic effect in this olaparib-sensitive model. The results showed that the relative tumor inhibition rate (TGI) (%) in the combination group was 100%, which was statistically significant compared to the control group (p<0.001).

[0144] We found it difficult to distinguish differences in the efficacy of olaparib alone, TH-302 alone, and the combination groups based on relative tumor inhibition (TGI) results. However, our study showed that both TH-302 alone and in combination significantly improved tumor clearance compared to olaparib alone. Results showed that none of the six mice treated with olaparib alone achieved complete tumor clearance. However, complete tumor clearance was observed in one mouse treated with 40 mg / kg TH-302 and in six mice treated with 80 mg / kg TH-302 alone, for clearance rates of 16.7% and 100%, respectively. Complete tumor clearance was also observed in the combination of 40 mg / kg TH-302 and 50 mg / kg olaparib. Only one mouse achieved complete tumor clearance with 40 mg / kg TH-302, while no mouse achieved complete tumor clearance with olaparib alone. However, in the combination group, all six mice achieved complete tumor clearance at the same dose, for a 100% clearance rate. Statistical analysis of TGI in each group of mice showed that the TH-302 80mg / kg group exhibited significantly greater inhibitory effects on tumor proliferation compared to the Olaparib 50mg / kg group. Furthermore, the combination of the test drug TH-302 40mg / kg and Olaparib 50mg / kg demonstrated statistically significant superior tumor inhibition compared to the Oplaparib 50mg / kg and TH-302 40mg / kg monotherapy groups, demonstrating a synergistic anti-tumor effect between TH-302 and Olaparib.

[0145] The above results all indicate that:

[0146] 1. In the Olaparib-sensitive ovarian cancer OV5304 PDX model, both TH-302 alone and in combination with olaparib showed significantly superior tumor clearance and anti-tumor proliferation abilities compared to olaparib alone.

[0147] 2. Further comparison showed that TH-302 monotherapy was more effective than the PAPR inhibitor Olaparib alone in treating BRCA-mutated cancer models;

[0148] 3. In the experiment, it was observed that the efficacy of TH-302 combined with the PAPR inhibitor Olaparib was better than that of each single drug treatment.

[0149] In summary, in terms of efficacy, even in an Olaparib-sensitive experimental model, TH-302 alone was significantly more effective than the PARP inhibitor Olaparib, and its combination with Olaparib was even more effective.

[0150] In terms of safety, mice in the test drug TH-302 and Olaparib single-drug and combination treatment groups did not show any obvious weight loss, and were well tolerated during the treatment period. This preliminarily shows that the combination of the two drugs has a certain safety at a certain dosage level.

[0151] 2. Efficacy and safety evaluation of TH-302 in the Olaparib-resistant Capan-1 CDX pancreatic cancer model

[0152] Protocol: BALB / c nude mice were subcutaneously inoculated with human pancreatic cancer Capan-1 cells to establish a subcutaneous human pancreatic cancer transplantation model.

[0153] The experiment was divided into 7 groups, with 6 mice in each group, including the test drug Olaparib 100 mg / kg single drug group (Group 2), TH-302 50 mg / kg single drug group (Group 3, QD), TH-302 100 mg / kg single drug group (Group 4, QW), TH-302 50 mg / kg single drug group (Group 5, QW), TH-302 25 mg / kg single drug group (Group 6, QW), TH-302 25 mg / kg and Olaparib 100 mg / kg combined administration group (Group 7), and 10% anhydrous ethanol + 10% polyoxyethylene (35) castor oil + 80% glucose injection D5W (pH 7.4) vehicle control group (Group 1). The vehicle control group and each test drug TH-302 were administered by tail vein injection. Among them, the TH-302 50mg / kg monotherapy group (Group 3, QD) was administered once a day for 3 consecutive days, rested for 4 days, and then rested for two weeks; then administered every day for 3 consecutive days. The TH-302 100mg / kg (Group 4, QW), 50mg / kg (Group 5, QW), 25mg / kg (Group 6, QW) monotherapy groups and the TH-302 25mg / kg and Olaparib 100mg / kg combined administration group (Group 7) were all administered once a week for a total of three weeks. The test drug Olaparib in each group was administered orally by gavage once a day for a total of 30 days. The specific administration route, dosage and regimen in the human pancreatic cancer Capan-1 animal model are shown in Table 2.

[0154] Table 2: Administration routes, doses, and regimens in the human pancreatic cancer Capan-1 animal model Note: The administration volume is 10 μL / g.

[0155] Tumor growth was recorded for each treatment group and control group on different days of the experiment, as shown in Table 3. The corresponding tumor volume growth curves for each group are shown in Figure 3. Efficacy was evaluated based on relative tumor growth rate and relative tumor inhibition rate, and the efficacy analysis for each group is shown in Table 4. Post-dose body weight changes in the treatment and control groups were recorded to investigate the safety of each group in the human pancreatic cancer Capan-1 subcutaneous model. The body weight change rate of mice on day 43 is shown in Table 5, and the body weight change curves for each treatment group over time are shown in Figure 4.

[0156] Table 3: Changes in tumor volume of mice in each group with treatment time in the human pancreatic cancer Capan-1 model (mm 3 )

[0157] Table 4: Efficacy analysis of each group in the human pancreatic cancer Capan-1 subcutaneous model

[0158] Table 5: Body weight changes of each group in the human pancreatic cancer Capan-1 subcutaneous model (43 days after inoculation)

[0159] The above data show that:

[0160] The average tumor volume of mice in the vehicle control group was 1301.38 mm on Day 43 after tumor cell inoculation. 3 The average tumor volume of the test drug Olaparib 100mg / kg (Group 2) treatment group on Day 43 after tumor cell inoculation was 846.86mm 3 The relative tumor inhibition rate TGI (%) was 37.43%, which had no statistically significant difference compared with the control group (p>0.05).

[0161] In the test drug TH-302 50 mg / kg treatment group (Group 3, QD), the average tumor volume was 146.99 mm on Day 43 after tumor cell inoculation. 3 Compared with the control group, there was a statistically significant difference (p<0.001), with a relative tumor inhibition rate (TGI) of 89.17%. In the test drug TH-302 100mg / kg treatment group (Group 4, QW), the average tumor volume on Day 43 after tumor cell inoculation was 124.68mm 3Compared with the control group, there was a statistically significant difference (p<0.001), and the relative tumor inhibition rate (TGI) was 90.89%. The test drug TH-302 50mg / kg treatment group (Group 5, QW) had an average tumor volume of 263.45mm on Day 43 after tumor cell inoculation. 3 Compared with the control group, there was a statistically significant difference (p<0.001), with a relative tumor inhibition rate (TGI) of 80.18% and a complete tumor inhibition rate of 33.3%. In the test drug TH-302 25mg / kg treatment group (Group 6, QW), the average tumor volume on Day 43 after tumor cell inoculation was 1521.33mm 3 There was no statistically significant difference compared to the control group (p>0.05), and the relative tumor inhibition rate (TGI) was -21.59%. In the Olaparib 100mg / kg and TH-302 25mg / kg combination treatment group (Group 7), the average tumor volume was 378.56mm on Day 43 after tumor cell inoculation. 3 There was a statistically significant difference compared with the control group (p<0.001), and the relative tumor inhibition rate TGI (%) was 74.36%.

[0162] TH-302 monotherapy groups at 100 mg / kg (Group 4, QW), 50 mg / kg (Group 5, QW), and 25 mg / kg (Group 6, QW) demonstrated dose-dependent antitumor effects. Statistically significant differences were observed between the 100 mg / kg (Group 4, QW) and 50 mg / kg (Group 5, QW) monotherapy groups and the 25 mg / kg (Group 6, QW) monotherapy group (p < 0.001).

[0163] The anti-tumor effect of the combination treatment group of olaparib 100mg / kg and TH-302 25mg / kg (Group 7) was superior to that of the monotherapy group of olaparib 100mg / kg (Group 2) and TH-302 25mg / kg (Group 6), with statistically significant differences (p<0.05 and p<0.001). The specific dose combination of olaparib and TH-302 can show a synergistic effect in tumor inhibition.

[0164] There was no significant weight loss in mice treated with the test drugs Olaparib 100 mg / kg (Group 2), TH-302 25 mg / kg (Group 6), TH-302 50 mg / kg (Group 3, QD), TH-302 50 mg / kg (Group 5, QW), TH-302 100 mg / kg (Group 4, QW), Olaparib 100 mg / kg and TH-302 25 mg / kg combined group (Group 7), and vehicle control group (Group 1), indicating good tolerance.

[0165] The above research results show that:

[0166] 1. A specific dose of TH-302 monotherapy demonstrated excellent anti-tumor efficacy in pancreatic cancer models resistant to olaparib (the statistical average TGI in six experimental mice was 37.43%, which is generally judged by technicians to be around 60%, indicating a certain degree of resistance) (Groups 3, 4, and 5). However, a low-dose TH-302 monotherapy regimen had no anti-tumor effect (Group 6).

[0167] 2. Furthermore, the combination of low-dose TH-302 and Olaparib showed anti-tumor effects in Olaparib-resistant pancreatic cancer models (Group 7);

[0168] That is to say, for patients who are tolerant to Olaparib, or patients who can only receive low-dose TH-302 treatment (in this low-dose treatment regimen, tumor growth is poorly inhibited or cannot be effectively inhibited), the combination of TH-302 and Olaparib can achieve tumor treatment effects, and the combination regimen will provide them with a new treatment option.

[0169] In summary, the combination of TH-302 and Olaparib will expand the use scenarios of the two drugs, allowing patients who do not have a significant therapeutic effect on the single-dose regimen of either drug to benefit from the treatment effect of the combination without significantly reducing safety.

[0170] Although the PARP inhibitor selected in the examples of the present application is Olaparib, Rucaparib, Niraparib, Talazoparib, Fluzoparib, Pamiparib, etc. are also PARP inhibitors, and their mechanism of action is similar to that of Olaparib, which is to block the enzymes involved in repairing damaged DNA. Therefore, it can be inferred that PARPi such as Rucaparib, Niraparib, Talazoparib, Fluzoparib, Pamiparib, etc. have similar tumor inhibitory effects as Olaparib in the above experiments.

[0171] TH-302 is a hypoxia-activated DNA alkylating agent, a compound of the general formula of claim 1:

[0172] The mechanism of action of these compounds has been demonstrated to be similar to that of TH-302 in relevant patent applications, so it is entirely predictable that these compounds will have similar effects to TH-302.

Claims

1. A method of treating cancer or tumor patients using a drug containing a hypoxia-activated compound of formula (I) alone or in combination with a PARP inhibitor: in, R is independently selected from H, -CH3, -CH2CH3, -CF3, and X is independently selected from leaving functional groups such as Cl, Br, MsO, TsO, etc.

2. The method of claim 1, wherein The patient has impaired DNA repair enzymes.

3. The method according to claim 1 or 2, wherein The patient's tumor or cancer tissue is detected to have any one or two pathogenic gene mutations in the genes corresponding to BRCA1 and BRCA2; or The patient is detected to have any one pathogenic gene mutation or two pathogenic gene mutations in the genes corresponding to BRCA1 and BRCA2.

4. The method of claim 3, wherein: BRCA1 and BRCA2 mutations include germline mutations (gBRCAm) and somatic mutations (sBRCAm) of BRCA1 and BRCA2 mutations.

5. The method of claim 1 or 2, wherein: The hypoxia-activated compound of formula (I) is selected from the compounds of the following structures:

6. The method of claim 1 or 2, wherein: The PARP inhibitor is selected from Olaparib, Rucaparib, Niraparib, Talazoparib, Fluzoparib, and Pamiparib.

7. The method of claim 1 or 2, wherein: The cancer or tumor is selected from ovarian cancer, breast cancer, pancreatic cancer, fallopian tube cancer, primary peritoneal cancer, gastric cancer, prostate cancer, non-small cell lung cancer, small cell lung cancer, liver cancer, colon cancer, and rectal cancer.

8. The method of claim 1 or 2, wherein: The cancer or tumor patient is sensitive or resistant to PARP inhibitors.

9. The method of claim 1 or 2, wherein the compound having the following structure is used as a monotherapy to treat cancer or tumor patients that are sensitive or resistant to PARP inhibitors:

10. The method of claim 1 or 2, wherein the compound having the following structure is used in combination with a PARP inhibitor to treat cancer or tumor patients who are sensitive or resistant to PARP inhibitors:

11. The method of claim 1, wherein: The TMB (tumor mutation burden) level of the gene mutation is medium.

12. A method of treatment comprising the following steps: Detect BRCA1 and BRCA2 gene mutations in cancer and tumor patients; If a patient is found to have a pathogenic gene mutation in one or both of the genes corresponding to BRCA1 and BRCA2, treatment may be performed using a drug containing a hypoxia-activated compound of formula (I) alone or in combination with a PARP inhibitor: Wherein, R is independently selected from H, -CH3, -CH2CH3, -CF3, and X is independently selected from leaving functional groups such as Cl, Br, MsO, and TsO.

13. A unit package of a pharmaceutical preparation, comprising an independent packaging container containing a pharmaceutical preparation, an outer packaging component for accommodating the independent packaging container, and a drug instruction sheet, characterized in that: The pharmaceutical preparation contains a hypoxia-activated compound of formula (I), and the drug instructions state that the drug is used in combination with a PARP inhibitor to treat cancer and tumor patients: Wherein, R is independently selected from H, -CH3, -CH2CH3, -CF3, and X is independently selected from leaving functional groups such as Cl, Br, MsO, TsO, etc. Preferably, the patient's DNA repair enzyme is impaired, Preferably, the patient's tumor or cancer tissue is detected to have any one or two pathogenic gene mutations in the genes corresponding to BRCA1 and BRCA2; or the patient is detected to have any one or two pathogenic gene mutations in the genes corresponding to BRCA1 and BRCA2, Preferably, BRCA1 and BRCA2 mutations include germline mutations (gBRCAm) and somatic mutations (sBRCAm) of BRCA1 and BRCA2 mutations. Preferably, the hypoxia-activated compound of formula (I) is selected from the compounds of the following structures: Preferably, the PARP inhibitor is selected from Olaparib, Rucaparib, Niraparib, Talazoparib, Fluzoparib, Pamiparib, Preferably, the cancer or tumor is selected from ovarian cancer, breast cancer, pancreatic cancer, fallopian tube cancer, primary peritoneal cancer, gastric cancer, prostate cancer, non-small cell lung cancer, small cell lung cancer, liver cancer, colon cancer, and rectal cancer. Preferably, the cancer or tumor patient is sensitive or resistant to PARP inhibitors.

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