Combination of POLθ inhibitor and PARP inhibitor and use thereof

Abstract

A combination of a POLθ inhibitor and a PARP inhibitor, having a synergistic inhibitory effect in the treatment of tumors.

Description

A combination of POLθ inhibitor and PARP inhibitor and its application

[0001] This application claims priority to Chinese patent application 2024112105506, filed on August 30, 2024. The entire contents of the aforementioned Chinese patent application are incorporated herein by reference. Technical Field

[0002] This invention belongs to the field of biomedical technology, specifically relating to a combination of a POLθ inhibitor and a PARP inhibitor and their application. Background Technology

[0003] PARP inhibitors are cancer therapies targeting poly-ADP-ribose polymerase. They were the first anticancer drugs to be approved for clinical use using the concept of synthetic lethality. Currently approved and investigational drugs include olaparib, niraparib, tapazoli, pamiparib, rucaparib, fluzoparib, sennaparib, and saruparib (AZD5305). However, clinical observations have shown that tumors can develop resistance to PARP inhibitors through various mechanisms, and the hematologic toxicity and other side effects of PARP itself limit its clinical value. Therefore, there is an urgent need for combination therapies that can synergistically enhance efficacy and reduce toxicity.

[0004] POLθ (Polymerase theta) is a member of the A family of polymerases, encoding a protein of approximately 290 kDa. Its unique structure comprises an N-terminal helicase-like domain and a C-terminal DNA polymerase domain, with an amorphous region in between. This structure endows POLθ with special biochemical properties. POLθ is known to promote alternative non-homologous end joining (alt-NHEJ) repair pathways. In cancers with BRCA1 / 2 deficiency or reduced homologous recombination (HR) repair capacity, cells rely more heavily on the alt-NHEJ pathway for DNA repair. On the other hand, single-stranded DNA gaps generated after cell replication are repaired by homologous recombination pathways involving proteins such as BRCA1 / 2. In cells with BRCA1 / 2 gene mutations or loss of function, this repair mechanism is impaired, leading to the accumulation of DNA damage. When these cells are treated with PARP inhibitors, the damage is further aggravated. To address this damage, cells rely on the POLθ-mediated end-joining repair pathway to attempt to close these gaps. Therefore, POLθ inhibitors and PARP inhibitors have the potential for synergistic effects when used in combination in cells with impaired homologous recombination repair function. Summary of the Invention

[0005] To address the technical problem of the lack of effective combination therapy for cancer treatment using POLθ inhibitors and PARP inhibitors in existing technologies, this invention provides a combination of POLθ inhibitors and PARP inhibitors and its application. This combination has demonstrated good synergistic inhibition in multiple solid tumor models, exhibiting superior and more durable therapeutic effects compared to other PARP inhibitor-based combination therapies. Furthermore, it maintains good and durable therapeutic effects even when the PARP inhibitor dose is reduced to half the clinical dose, demonstrating potential for synergistic enhancement and toxicity reduction.

[0006] The present invention solves the above-mentioned technical problems through the following technical solutions.

[0007] A first aspect of the present invention provides a combination of a POLθ inhibitor and a PARP inhibitor, the combination comprising a POLθ inhibitor and a PARP inhibitor; the POLθ inhibitor comprising one or more compounds represented by formulas 1-3 below and pharmaceutically acceptable salts thereof:

[0008] In a preferred embodiment, the PARP inhibitor is selected from one or more of olaparib, niraparib, talazoparib, pamiparib, rucaparib, fluzoparib, senaparib, saruparib (AZD5305), and their pharmaceutically acceptable salts, and the specific structures of these PARP inhibitors are as follows:

[0009] In a preferred embodiment, the pharmaceutically acceptable salt of the POLθ inhibitor is selected from one or more of hydrochloride, sulfate, trifluoroacetate, formate, and p-toluenesulfonate.

[0010] In a preferred embodiment, the mass ratio of the POLθ inhibitor to the PARP inhibitor is 1:(0.001-1) or 0.5:(1-300), for example 1:0.002, 1:0.6, 1:1, 1:2, 1:20, 1:50, 1:100 or 1:200.

[0011] In a preferred embodiment, the combination comprises one or more of the compounds shown in Formulas 1-3 above, and one or more PARP inhibitors selected from the following: olaparib, niraparib, and saruparib.

[0012] In one preferred embodiment, the combination comprises the compound shown in Formula 3 and olaparib;

[0013] The combination includes the compound shown in Formula 1 and olaparib;

[0014] The combination includes the compound shown in Formula 1 and niraparib;

[0015] The combination comprises the compound shown in Formula 1 and Saruparib; or

[0016] The combination includes the compound shown in Formula 2 and olaparib.

[0017] A second aspect of the invention provides the use of the combination as described in the first aspect of the invention in the preparation of a medicament for treating and / or preventing tumors.

[0018] In a preferred embodiment, the tumor includes one or more of the following: endometrial cancer, lung cancer, breast cancer, gastric cancer, ovarian cancer, prostate cancer, colorectal cancer, pancreatic cancer, uterine leiomyosarcoma, multiple myeloma, cervical cancer, urinary tract-related cancer, skin cancer, and kidney cancer. For example, the tumor is endometrial cancer, breast cancer, gastric cancer, ovarian cancer, colorectal cancer, or pancreatic cancer.

[0019] In a preferred embodiment, the tumor includes one or more of the following: tumors deficient in homologous recombination repair genes, tumors deficient in TLS (translesion synthesis) polymerase, tumors deficient in the Shieldin complex, and tumors with high POLθ expression.

[0020] In a preferred embodiment, the tumor includes a homologous recombination repair gene-deficient tumor and / or a TLS polymerase-deficient tumor.

[0021] In a preferred embodiment, the homologous recombination repair gene-deficient tumor is a tumor that is positive as assessed by the HRD assessment system. The score of the HRD assessment system is calculated based on the unweighted sum of scores of three ultrastructural features: large segment migration (LST), telomere allele imbalance (TAI), and loss of heterozygosity (LOH). See Moore GM, et al, Examining the preyalence of homologous recombination repair defects in ER+ breast cancers. Breast Cancer Res Treat. 2022 Apr; 192(3):649-653.

[0022] In a preferred embodiment, the homologous recombination repair gene defect is a gene defect that causes HRD (homologous recombination deficiency) in cells, preferably including one or more of the following target genes with decreased expression and / or abnormal protein function: BRCA1, BRCA2, PALB2, RAD51, RAD51C, RAD51B, RAD51D, SLX4, FANCD2, FANCF, and XRCC3; preferably, the homologous recombination repair gene defect includes one or more of the following target genes with decreased expression and / or abnormal protein function: BRCA1, BRCA2, RAD51C, and XRCC3; more preferably, the homologous recombination repair gene defect includes one or more of the following target genes with decreased expression and / or abnormal protein function: BRCA1, BRCA2, RAD51C, and XRCC3.

[0023] In a preferred embodiment, the TLS polymerase gene defect includes one or more of the following target genes with decreased expression and / or abnormal protein function: POLI, POLH, RAD18, REV1, REV3, POLK, POLL, PCNA, and REV7; preferably, the TLS polymerase defect includes one or more of the following target genes with decreased expression and / or abnormal protein function: REV7, POLE2, POLH, and REV3; more preferably, the TLS polymerase gene defect includes one or two of the following target genes with decreased expression and / or abnormal protein function: REV7 and REV3.

[0024] In a preferred embodiment, the Shieldin complex defect includes one or more of the following target genes with decreased expression and / or abnormal protein function: RIF1, 53BP1, SHLD2, SHLD1, SHLD3, and REV7; preferably, the Shieldin complex defect includes one or two of the following target genes with decreased expression and / or abnormal protein function: REV7, RIF1, 53BP1, and SHLD2.

[0025] In this invention, the decrease in expression includes the case of no expression.

[0026] In a preferred embodiment, the tumor includes one or more of the following: endometrial cancer with a deficiency in homologous recombination repair genes, lung cancer with a deficiency in homologous recombination repair genes, breast cancer with a deficiency in homologous recombination repair genes, gastric cancer with a deficiency in homologous recombination repair genes, ovarian cancer with a deficiency in homologous recombination repair genes, prostate cancer with a deficiency in homologous recombination repair genes, colorectal cancer with a deficiency in homologous recombination repair genes, pancreatic cancer with a deficiency in homologous recombination repair genes, uterine leiomyosarcoma with a deficiency in homologous recombination repair genes, multiple myeloma with a deficiency in homologous recombination repair genes, cervical cancer with a deficiency in homologous recombination repair genes, urinary tract-related cancer with a deficiency in homologous recombination repair genes, skin cancer with a deficiency in homologous recombination repair genes, and renal cancer with a deficiency in homologous recombination repair genes. Preferably, the tumor further includes a deficiency in the Shieldin complex (e.g., SHLD2 deficiency).

[0027] In a preferred embodiment, the tumor includes TLS polymerase-deficient endometrial cancer, TLS polymerase-deficient lung cancer, TLS polymerase-deficient breast cancer, TLS polymerase-deficient gastric cancer, TLS polymerase-deficient ovarian cancer, TLS polymerase-deficient prostate cancer, TLS polymerase-deficient colorectal cancer, TLS polymerase-deficient pancreatic cancer, TLS polymerase-deficient uterine leiomyosarcoma, TLS polymerase-deficient multiple myeloma, TLS polymerase-deficient cervical cancer, TLS polymerase-deficient urinary tract-related cancer, TLS polymerase-deficient skin cancer, or TLS polymerase-deficient kidney cancer.

[0028] In a preferred embodiment, the homologous recombination repair gene-deficient tumor is a homologous recombination repair gene-deficient ovarian cancer, including homologous recombination repair gene-deficient primary ovarian cancer and homologous recombination repair gene-deficient metastatic ovarian cancer. The homologous recombination repair gene-deficient ovarian cancer may include one or more of BRCA2 deficiency, BRCA1 deficiency, XRCC3 deficiency, RAD51B deficiency, and RAD51C deficiency. Preferably, the homologous recombination repair gene-deficient ovarian cancer further includes one or more of RIF1 deficiency, SHLD2 deficiency, and 53BP1 deficiency.

[0029] In a preferred embodiment, the homologous recombination repair gene-deficient tumor is homologous recombination repair gene-deficient lung cancer, including homologous recombination repair gene-deficient primary lung cancer and homologous recombination repair gene-deficient metastatic lung cancer. The homologous recombination repair gene-deficient lung cancer may include one or more of BRCA2 deficiency, BRCA1 deficiency, XRCC3 deficiency, RAD51B deficiency, and RAD51C deficiency. Preferably, the homologous recombination repair gene-deficient lung cancer further includes one or more of RIF1 deficiency, SHLD2 deficiency, REV7 deficiency, and 53BP1 deficiency.

[0030] In a preferred embodiment, the homologous recombination repair gene-deficient tumor is a homologous recombination repair gene-deficient breast cancer, including homologous recombination repair gene-deficient primary breast cancer and homologous recombination repair gene-deficient metastatic breast cancer. The homologous recombination repair gene-deficient breast cancer may include one or more of BRCA2 deficiency, BRCA1 deficiency, XRCC3 deficiency, RAD51B deficiency, and RAD51C deficiency. Preferably, the homologous recombination repair gene-deficient breast cancer further includes one or more of RIF1 deficiency, SHLD2 deficiency, REV7 deficiency, and 53BP1 deficiency.

[0031] In a preferred embodiment, the homologous recombination repair gene-deficient tumor is homologous recombination repair gene-deficient gastric cancer, including homologous recombination repair gene-deficient primary gastric cancer and homologous recombination repair gene-deficient metastatic gastric cancer. The homologous recombination repair gene-deficient gastric cancer may include one or more of BRCA2 deficiency, BRCA1 deficiency, XRCC3 deficiency, RAD51B deficiency, and RAD51C deficiency. Preferably, the homologous recombination repair gene-deficient gastric cancer further includes one or more of RIF1 deficiency, SHLD2 deficiency, REV7 deficiency, and 53BP1 deficiency.

[0032] In a preferred embodiment, the homologous recombination repair gene-deficient tumor is homologous recombination repair gene-deficient prostate cancer, including homologous recombination repair gene-deficient primary prostate cancer and homologous recombination repair gene-deficient metastatic prostate cancer. The homologous recombination repair gene-deficient prostate cancer may include one or more of BRCA2 deficiency, BRCA1 deficiency, XRCC3 deficiency, RAD51B deficiency, and RAD51C deficiency. Preferably, the homologous recombination repair gene-deficient prostate cancer further includes one or more of RIF1 deficiency, SHLD2 deficiency, REV7 deficiency, and 53BP1 deficiency.

[0033] In a preferred embodiment, the homologous recombination repair gene-deficient tumor is homologous recombination repair gene-deficient colorectal cancer, including homologous recombination repair gene-deficient primary colorectal cancer and homologous recombination repair gene-deficient metastatic colorectal cancer. The homologous recombination repair gene-deficient colorectal cancer may include one or more of BRCA2 deficiency, BRCA1 deficiency, XRCC3 deficiency, RAD51B deficiency, and RAD51C deficiency. Preferably, the homologous recombination repair gene-deficient colorectal cancer further includes one or more of RIF1 deficiency, SHLD2 deficiency, REV7 deficiency, and 53BP1 deficiency.

[0034] In a preferred embodiment, the homologous recombination repair gene-deficient tumor is homologous recombination repair gene-deficient pancreatic cancer, including homologous recombination repair gene-deficient primary pancreatic cancer and homologous recombination repair gene-deficient metastatic pancreatic cancer. The homologous recombination repair gene-deficient pancreatic cancer may include one or more of BRCA2 deficiency, BRCA1 deficiency, XRCC3 deficiency, RAD51B deficiency, and RAD51C deficiency. Preferably, the homologous recombination repair gene-deficient pancreatic cancer further includes one or more of RIF1 deficiency, SHLD2 deficiency, REV7 deficiency, and / or 53BP1 deficiency.

[0035] In a preferred embodiment, the homologous recombination repair gene-deficient tumor is a homologous recombination repair gene-deficient uterine leiomyosarcoma, including primary uterine leiomyosarcoma and metastatic uterine leiomyosarcoma. The homologous recombination repair gene-deficient uterine leiomyosarcoma may further include one or more of BRCA2 deficiency, BRCA1 deficiency, XRCC3 deficiency, RAD51B deficiency, and RAD51C deficiency. Preferably, the homologous recombination repair gene-deficient uterine leiomyosarcoma further includes one or more of RIF1 deficiency, SHLD2 deficiency, REV7 deficiency, and 53BP1 deficiency.

[0036] In a preferred embodiment, the homologous recombination repair gene-deficient tumor is a homologous recombination repair gene-deficient multiple myeloma, including homologous recombination repair gene-deficient primary multiple myeloma and homologous recombination repair gene-deficient metastatic multiple myeloma. The homologous recombination repair gene-deficient multiple myeloma may further include one or more of BRCA2 deficiency, BRCA1 deficiency, XRCC3 deficiency, RAD51B deficiency, and RAD51C deficiency. Preferably, the homologous recombination repair gene-deficient multiple myeloma further includes one or more of RIF1 deficiency, SHLD2 deficiency, REV7 deficiency, and 53BP1 deficiency multiple myeloma.

[0037] In a preferred embodiment, the TLS polymerase-deficient cancers include TLS polymerase-deficient primary cancers and TLS polymerase-deficient metastatic cancers, such as TLS polymerase-deficient primary lung cancer, TLS polymerase-deficient metastatic lung cancer, TLS polymerase-deficient primary breast cancer, TLS polymerase-deficient metastatic breast cancer, TLS polymerase-deficient primary gastric cancer, TLS polymerase-deficient metastatic gastric cancer, TLS polymerase-deficient primary ovarian cancer, TLS polymerase-deficient metastatic ovarian cancer, TLS polymerase-deficient primary prostate cancer, TLS polymerase-deficient metastatic prostate cancer, TLS polymerase-deficient primary colorectal cancer, TLS polymerase-deficient metastatic colorectal cancer, TLS polymerase-deficient primary pancreatic cancer, TLS polymerase-deficient metastatic pancreatic cancer, TLS polymerase-deficient primary uterine leiomyosarcoma, TLS polymerase-deficient metastatic uterine leiomyosarcoma, TLS polymerase-deficient multiple primary myeloma, or TLS polymerase-deficient multiple metastatic myeloma.

[0038] In a preferred embodiment, the homologous recombination repair gene-deficient cancers include primary cancers and metastatic cancers, such as primary lung cancer, metastatic lung cancer, primary breast cancer, metastatic breast cancer, primary gastric cancer, metastatic gastric cancer, primary ovarian cancer, and metastatic ovarian cancer. Cancer, homologous recombination repair gene-deficient metastatic prostate cancer, homologous recombination repair gene-deficient primary prostate cancer, homologous recombination repair gene-deficient primary colorectal cancer, homologous recombination repair gene-deficient metastatic colorectal cancer, homologous recombination repair gene-deficient primary pancreatic cancer, homologous recombination repair gene-deficient metastatic pancreatic cancer, homologous recombination repair gene-deficient primary uterine leiomyosarcoma, homologous recombination repair gene-deficient metastatic uterine leiomyosarcoma, homologous recombination repair gene-deficient multiple primary myeloma or homologous recombination repair gene-deficient multiple metastatic myeloma.

[0039] In a preferred embodiment, the homologous recombination repair gene-deficient tumor is a homologous recombination repair gene-deficient endometrial cancer, including homologous recombination repair gene-deficient primary endometrial cancer and homologous recombination repair gene-deficient metastatic endometrial cancer. The homologous recombination repair gene-deficient endometrial cancer may further include one or more of BRCA2 deficiency, BRCA1 deficiency, XRCC3 deficiency, RAD51B deficiency, and RAD51C deficiency. Preferably, the homologous recombination repair gene-deficient endometrial cancer further includes one or more of RIF1 deficiency, SHLD2 deficiency, REV7 deficiency, and 53BP1 deficiency.

[0040] In a preferred embodiment, the Shieldin complex-deficient tumor includes one or more of the following: Shieldin complex-deficient endometrial cancer, Shieldin complex-deficient lung cancer, Shieldin complex-deficient breast cancer, Shieldin complex-deficient gastric cancer, Shieldin complex-deficient ovarian cancer, Shieldin complex-deficient prostate cancer, Shieldin complex-deficient colorectal cancer, Shieldin complex-deficient pancreatic cancer, Shieldin complex-deficient uterine leiomyosarcoma, Shieldin complex-deficient multiple myeloma, Shieldin complex-deficient cervical cancer, Shieldin complex-deficient urinary tract-related cancer, Shieldin complex-deficient skin cancer, and Shieldin complex-deficient renal cancer; preferably, the Shieldin complex-deficient tumor includes SHLD2-deficient tumors and / or RIF1-deficient tumors.

[0041] In a preferred embodiment, the homologous recombination repair gene-deficient cancer is BRCA2-deficient colorectal cancer, BRCA2-deficient primary colon cancer, BRCA2-deficient prostate cancer, RAD51B-deficient, POLE2-deficient, and POLH-deficient metastatic prostate cancer (preferably, the metastatic prostate cancer contains RAD51B, POLB, POLE2, POLH, MSH6, and RNASEH2B deficiencies), BRCA2-deficient ovarian cancer, BRCA2-deficient uterine leiomyosarcoma, BRCA-deficient breast cancer, XRCC3-deficient colon cancer, RAD51C-deficient colon cancer, 53BP1-deficient breast cancer, BRCA2-deficient and SHLD2-deficient breast cancer, 53BP1-deficient and SHLD2-deficient breast cancer, BRCA1-deficient and SHLD2-deficient breast cancer, FANCF-deficient ovarian cancer, POLθ-overexpressing gastric cancer, BRCA2-deficient and FANCD2-deficient multiple myeloma, or TLS polymerase-deficient colon cancer.

[0042] In a preferred embodiment, the tumor is BRCA2-deficient endometrial cancer, BRCA1-deficient breast cancer, BRCA1-deficient and SHLD2-deficient breast cancer, BRCA2-deficient gastric cancer, RAD51C-deficient and RIF1-deficient gastric cancer, BRCA2-deficient colorectal cancer, or BRCA2-deficient pancreatic cancer.

[0043] A third aspect of the present invention provides a medicament for treating and / or preventing tumors, said medicament comprising a POLθ inhibitor and a PARP inhibitor;

[0044] The POLθ inhibitor and PARP inhibitor are defined as combinations as described in the first aspect of the invention;

[0045] The tumor is defined as the application described in the second aspect of the present invention.

[0046] In a preferred embodiment, the mass ratio of the POLθ inhibitor to the PARP inhibitor in the drug is 1:(0.001-1) or 0.5:(1-300), for example 1:0.002, 1:0.6, 1:1, 1:2, 1:20, 1:50, 1:100 or 1:200.

[0047] In a preferred embodiment, the drug can be formulated into various suitable dosage forms depending on the route of administration, including gastrointestinal dosage forms (e.g., oral dosage forms) and non-gastrointestinal dosage forms (e.g., injection dosage forms, respiratory dosage forms, mucosal dosage forms, or cavity dosage forms), such as capsules, tablets, pills, granules, powders, or oral liquids, with tablets being preferred.

[0048] In a preferred embodiment, the drug is administered in a manner conventional to such drugs in the art, such as by injection (e.g., intravenous injection, subcutaneous injection, peritoneal injection or intramuscular injection) or oral administration, preferably oral administration.

[0049] A fourth aspect of the present invention provides a pillbox set, the pillbox set comprising pillbox A and pillbox B, wherein:

[0050] The medicine box A contains a POLθ inhibitor, which is defined as the combination described in the first aspect of the present invention;

[0051] The cassette B contains a PARP inhibitor, which is defined as the combination described in the first aspect of the invention.

[0052] In this invention, the "A" and "B" in medicine box A and medicine box B are only used for distinction and do not have any actual meaning.

[0053] The fifth aspect of the present invention provides the use of POLθ inhibitors in the preparation of medicaments that enhance the efficacy of PARP inhibitors;

[0054] The POLθ inhibitor or PARP inhibitor is defined as a combination as described in the first aspect of the invention.

[0055] In this invention, "enhancing the efficacy of PARP inhibitors" may refer to achieving the same or better efficacy with a smaller amount of PARP inhibitor (e.g., for mice, the minimum dose may be 25 mg / kg), or POLθ inhibitors that make PARP inhibitors more effective in applications where PARP inhibitors alone are ineffective.

[0056] In a preferred embodiment, the PARP inhibitor is effective in inhibiting the function of the PARP enzyme; further, it can be used for the prevention and / or treatment of PARP-related tumors, or for non-diagnostic or therapeutic purposes to inhibit the PARP enzyme (e.g., in vitro pharmacological experiments). Preferably, the tumor is defined as the application described in the second aspect of the invention.

[0057] A sixth aspect of the invention provides a combination as described in the first aspect of the invention for preparing a treatment and / or prevention of tumors, said tumors as defined by the application described in the second aspect of the invention.

[0058] A seventh aspect of the present invention provides a method for treating tumors, the method comprising administering a therapeutically effective amount of a POLθ inhibitor and a PARP inhibitor to a patient in need; or, the method comprising administering a therapeutically effective amount of a POLθ inhibitor to a patient in need of receiving PARP inhibitor treatment.

[0059] The POLθ inhibitor or PARP inhibitor is defined as a combination as described in the first aspect of the invention;

[0060] The tumor is defined as the application described in the second aspect of the present invention.

[0061] The eighth aspect of the present invention provides a method for enhancing the therapeutic effect of a PARP inhibitor, the method comprising administering to a patient in need a dose of a PARP inhibitor that effectively enhances the therapeutic effect of the PARP inhibitor;

[0062] The POLθ inhibitor or PARP inhibitor is defined as a combination as described in the first aspect of the invention.

[0063] In a preferred embodiment, the PARP inhibitor has the effect of preventing and / or treating tumors; preferably, the tumor is defined as the application described in the second aspect of the invention.

[0064] In one preferred embodiment, the dosage of the POLθ inhibitor is 50-600 mg / day, calculated based on the human and mouse species and dosage type; the dosage of the PARP inhibitor is 10-500 mg / day, calculated based on the human and mouse species and dosage type, for example, the dosage of olaparib can be 121.6-486.4 mg / day, and the dosage of niraparib can be 146 mg / day. The conversion of human and mouse species and dosage type is based on the industry guidance of the U.S. Food and Drug Administration (FDA), "Estimating the Maximum Safe Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers". In another preferred embodiment, the dosage of the PARP inhibitor can also be calculated based on the effective exposure to the target (see Clin Cancer Res 2022; 28:4724–36; AACR2024 CT014), for example, the dosage of AZD5305 can be 10 mg / day. Those skilled in the art can adjust the dosage appropriately according to the patient's condition (symptoms, number of treatments, intervals, etc.).

[0065] Terminology Explanation

[0066] The term "pharmaceutical acceptable" means that something is relatively non-toxic, safe, and suitable for patient use.

[0067] The term "pharmaceutically acceptable salt" refers to a salt obtained by reacting a compound with a pharmaceutically acceptable acid or base. When a compound contains a relatively acidic functional group, a base addition salt can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable base in a suitable inert solvent. Pharmaceutically acceptable base addition salts include, but are not limited to, sodium, potassium, calcium, aluminum, magnesium, bismuth, and ammonium salts. When a compound contains a relatively basic functional group, an acid addition salt can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable acid in a suitable inert solvent. Pharmaceutically acceptable acid addition salts include, but are not limited to, hydrochloride, sulfate, trifluoroacetate, formate, and p-toluenesulfonate. For details, see Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl, Camille G. Wermuth, 2011, 2nd Revised Edition).

[0068] The reagents and raw materials used in this invention are all commercially available.

[0069] The positive and progressive effects of this invention are as follows:

[0070] The combination of POLθ inhibitor and PARP inhibitor provided by this invention has good synergistic inhibition in the treatment of tumors, and can exert a better and more durable therapeutic effect than other combination therapies based on PARP inhibitors. It has high safety and has a promising prospect for the treatment of tumors. Attached Figure Description

[0071] Figure 1 shows the effects of compound 3 and olaparib in DLD1 BRCA2. - / - Results of anti-tumor cell viability assay on colorectal cancer cell lines.

[0072] Figure 2 shows the results of the anti-tumor cell viability test of compound 3 and olaparib on the PEO1 ovarian cancer cell line.

[0073] Figure 3 shows the results of the antitumor cell viability assay of compound 1 and Saruparib (AZD5305) on the gastric cancer cell line SNU601.

[0074] Figure 4 shows a crystal violet staining image of the inhibition of colony formation in the breast cancer cell line HCC1937 by compound 1 and olaparib.

[0075] Figure 5 shows the colony formation inhibition rates of compound 1 and olaparib on the breast cancer cell line HCC1937.

[0076] Figure 6 shows a crystal violet staining image of the inhibition of clonogenicity of compound 1 and AZD5305 on the pancreatic cancer cell line CAPAN-1.

[0077] Figure 7 shows the synergistic effect of compound 1 and AZD5305 on the antitumor cell viability of the pancreatic cancer cell line CAPAN-1.

[0078] Figure 8A shows the inhibitory effect of compound 2 and olaparib on the subcutaneous tumor volume of human breast cancer MDA-MB-436 in nude mice.

[0079] Figure 8B shows the effects of compound 2 and olaparib on the body weight of nude mice bearing human breast cancer MDA-MB-436.

[0080] Figure 9A shows the inhibitory effect of compound 1, p-toluenesulfonate, and olaparib on the volume of subcutaneous xenografts of human endometrial cancer EN-11-0101NOD-SCID in mice.

[0081] Figure 9B shows the effects of compound 1, p-toluenesulfonate, and olaparib on body weight in NOD-SCID mice carrying human endometrial cancer EN-11-0101.

[0082] Figure 10A shows the inhibitory effects of compound 1, p-toluenesulfonate, and olaparib on the volume of subcutaneous xenografts of human gastric cancer ST-02-0393NOD-SCID in mice.

[0083] Figure 10B shows the effects of compound 1, p-toluenesulfonate, and olaparib on body weight in NOD-SCID mice bearing human gastric cancer ST-02-0393.

[0084] Figure 11A shows the effects of compound 1 hydrochloride and niraparib / AZD5305 on human colorectal cancer DLD1 BRCA2. - / - Inhibition effect on subcutaneous tumor volume in nude mice.

[0085] Figure 11B shows the effect of compound 1 hydrochloride and niraparib / AZD5305 on BRCA2 in human colorectal cancer DLD1. - / - The effect on body weight in nude mice.

[0086] Figure 12A shows the effect of compound 1, p-toluenesulfonate, on human colorectal cancer DLD1 BRCA2 with low-dose olaparib. - / - Inhibition effect on subcutaneous tumor volume in nude mice.

[0087] Figure 12B shows the effects of compound 1, p-toluenesulfonate, and low-dose olaparib on human colorectal cancer DLD1 BRCA2. - / - The effect on body weight in nude mice.

[0088] Figure 13A shows the inhibitory effect of compound 1, p-toluenesulfonate, on the volume of subcutaneous xenografts of human breast cancer BR-05-0483E NOD-SCID mice compared with olaparib.

[0089] Figure 13B shows the effects of compound 1, p-toluenesulfonate, and olaparib on body weight in NOD-SCID mice bearing human breast cancer BR-05-0483E. Detailed Implementation

[0090] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0091] The structural formulas of POLθ inhibitor compounds 1, 2 and 3 used in the following examples are shown as Formulas 1, 2 and 3, respectively, and can be prepared according to the method disclosed in patent application 2023114104096 (see Examples 46, 87 and 216 of that patent application).

[0092] Example 1: Preparation of compound 1, p-toluenesulfonate

[0093] 30 mg of compound 1 (shown in Formula 1) and 10.44 mg of p-toluenesulfonic acid monohydrate were added to 0.5 mL of acetone / water (95:5, v:v), and the mixture was heated to 50 °C. The entire system was stirred at 50 °C for 2 hours, then cooled to 25 °C and stirred again. The mixture was filtered, and the resulting wet product was dried under vacuum at 50 °C for 2 hours to obtain the compound shown in Formula 4. NMR: 1H NMR (400MHz, DMSO-d6) δ 13.64 (s, 1H), 9.15 (d, J = 2.0Hz, 1H), 8.91 (s, 1H), 8.18 (s, 1H), 7.63 (s, 1H), 7.53 (s, 1H), 7.47 (d, J = 8.4Hz, 2H), 7.11 (d, J = 7.6Hz, 2H), 4.03 (s, 3H), 3.61 (s, 3H), 2.63 (s, 3H), 2.29 (s, 3H), 2.26 (d, J = 1.6Hz, 3H).

[0094] Example 2 Preparation of Compound 1 Hydrochloride

[0095] 25 L of ethyl acetate, 500 mL of methanol, and 200 g of free base of compound 1 were added to a reaction vessel. Stirring was started until the system was completely dissolved. 950 mL of a 4.0 N ethyl hydrochloride solution was added dropwise at room temperature. After the addition was complete, the system was stirred continuously at room temperature for 0.5 hours. Solid-liquid separation was then performed by filtration, and the filter cake was washed with 1 L of ethyl acetate. Finally, the filter cake was dried at 45–50 °C to obtain 201.6 g of the target compound, with a yield of 94.2%. NMR: 1H NMR (400MHz, DMSO-d6) δ 13.92 (s, 1H), 9.17 (d, J = 1.7Hz, 1H), 9.11 (s, 1H), 8.25 (s, 1H), 7.88 (s, 1H), 7.70 (s, 1H), 4.04 (s, 3H), 3.64 (s, 3H), 2.78 (s, 3H), 2.27 (d, J = 1.4Hz, 3H).

[0096] PARP inhibitors used in efficacy test cases

[0097] Olaparib (MedChemExpress, HY-10162 or Bidex Pharmaceuticals, BD162101)

[0098] Niraparib (Bide Pharmaceuticals, BD23406)

[0099] AZD5305 (MedchemExpress, HY-132167 or Bidex Pharmaceuticals, DQL796)

[0100] Cell lines and PDX used in the efficacy test cases

[0101] DLD1-BRCA2 - / - Cells: Hangzhou Shengyu Biomedical Technology Co., Ltd. established its own cells according to the literature (Ran, FA, Hsu, PD, Wright, J., Agarwala, V., Scott, DA, & Zhang, F. (2013). Genome engineering using the CRISPR-Cas9 system. Nature Protocols, 8(11), 2281-2308. doi.org / 10.1038 / nprot.2013.143). The parental cell line was derived from ATCC (CCL-221).

[0102] PEO1 is derived from ECACC(10032308);

[0103] SNU601 is derived from KCLB(00601);

[0104] HCC1937 is derived from ATCC (CRL-2336);

[0105] CAPAN1 is derived from ATCC (HTB-79);

[0106] MDA-MB-436 is derived from ATCC (HTB-130);

[0107] EN-11-0101, ST-02-0393, and BR-05-0483E are human tumor models established by WuXi AppTec.

[0108] Efficacy Test Example 1: Compound 3 and Olaparib in DLD1 BRCA2- / - Anti-tumor cell viability assay on colorectal cancer cell lines (BRCA2-deficient colon cancer cell lines)

[0109] Experimental Methods: The cell viability assay, commonly used in vitro, was performed using CellTiter Glo reagent (Promega, G7573) to assess the combined effect of compound 3 and olaparib, specifically the CTG assay. The specific assay method is as follows:

[0110] (1) Cultivating DLD1-BRCA2 - / -Cells were digested with trypsin (0.025% trypsin-EDTA, Hyclone) one day before the assay, centrifuged at 1000 rpm for 3 min, and collected. Cells were counted using a cell counter (Shanghai Mengwei Biomedical Technology Co., Ltd., SmartCell600A.SC1006) and seeded into 384-well cell culture plates at an appropriate seeding ratio (175 cells / well).

[0111] (2) Drug treatment was performed 24 hours after cell seeding, designated as day 0. An automated pipette (Thermo, Multidrop 8) was used to add the compounds. The plates were set to 384 wells, with an initial concentration of 50 μM for all compounds. Olapazone was diluted with DMSO, and 10 detection points were set (50 μM, 16.67 μM, 5.56 μM, 1.85 μM, 0.62 μM, 0.21 μM, 0.07 μM, 0.02 μM, 0.01 μM, and 0 μM). Compound 3 was diluted with DMSO, and 7 detection points were set (50 μM, 10 μM, 2 μM, 0.4 μM, 0.08 μM, 0.04 μM, and 0 μM). Two replicates were performed. Cells were cultured at 37°C in a 5% CO2 incubator.

[0112] (3) On day 7, remove the cell culture plate and add an equal volume of CTG reagent (the CTG reagent needs to be pre-mixed according to the reagent instructions to restore the temperature). Gently mix it for 10 minutes (speed 300) on a constant temperature mixer (Hangzhou Aosheng Instrument Co., Ltd., MSC-100). Perform chemiluminescence detection using an ELISA reader (Thermo, Varioskan LUX).

[0113] Data analysis: The coefficient of variation (CV%) of the control group test results should be less than 30%, and the z' value should be greater than 0.5. Data meeting the above quality control results were used to calculate cell viability (inhibition rate (%) = 100 × (control group average - experimental group) / (control group average - culture medium control group average). GraphPad Prism8 was used to perform nonlinear regression fitting of the compound's inhibition rate curve and derive the IC50. 50 value.

[0114] Experimental results: Compound 3 combined with olaparib showed a synergistic effect on olaparib at all concentrations (see Figure 1).

[0115] Example 2: Antitumor cell viability assay of compound 3 and olaparib in the PEO1 ovarian cancer cell line (BRCA2 mutant ovarian cancer cell line).

[0116] Experimental Methods: The cell viability assay, commonly used in vitro, was performed using CellTiter Glo reagent (Promega, G7573) to assess the combined effect of compound 3 and olaparib, specifically the CTG assay. The specific assay method is as follows:

[0117] (1) PEO1 cells were cultured. One day before the start of the assay, the cells were digested with trypsin (trypsin-EDTA, Hyclone), centrifuged at 1000 rpm for 3 min, and the cells were collected. The cells were counted using a counter (Shanghai Mengwei Biomedical Technology Co., Ltd., SmartCell600A.SC1006) and seeded into 384-well cell culture plates at an appropriate seeding ratio (300 cells / well).

[0118] (2) Drug treatment was performed 24 hours after cell seeding, marked as day 0. An automated pipette (Thermo, Multidrop 8) was used for compound addition. The plate was set to 384 wells. The initial concentration of olaparib was set to 50 μM, and it was diluted 1 / 3 with DMSO. Compound 3 was used to set four detection points (0 μM, 0.4 μM, 0.081 μM, and 0.016 μM), with two replicates. Cells were cultured at 37°C in a 5% CO2 incubator.

[0119] (3) On the 7th day, remove the cell culture plate and add an equal volume of CTG reagent (the CTG reagent needs to be restored to the temperature in advance according to the reagent instructions and premixed). Gently mix for 10 minutes (speed300) on a constant temperature mixer (Hangzhou Aosheng Instrument Co., Ltd., MSC-100). Use an ELISA reader (Thermo, Varioskan LUX) for chemiluminescence detection.

[0120] Data analysis: The CV% of the control group should be less than 30%, and the z' value should be greater than 0.5. Data meeting these quality control requirements were used to calculate cell viability (inhibition rate (%) = 100 × (control group average - experimental group) / (control group average - culture medium control group average). A nonlinear regression was used with GraphPad Prism8 to fit the inhibition rate curve of the compound and derive the IC50 value.

[0121] Experimental results: Compound 3, when used in combination with olaparib, showed a synergistic effect on olaparib at all concentrations (see Figure 2).

[0122] Example 3: Antitumor cell viability assay of compound 1 and Saruparib (AZD5305) in the gastric cancer cell line SNU601 (HRD, RAD51C methylated gastric cancer cell line).

[0123] Experimental Methods: The cell viability of compound 1 and AZD5305 in vitro was assessed using CellTiter Glo reagent (Promega, G7573), specifically the commonly used CTG assay. The specific assay method is as follows:

[0124] (1) SNU601 cells were cultured. One day before the assay, the cells were digested with trypsin (0.025% trypsin-EDTA, Hyclone), centrifuged at 1000 rpm for 3 min, and the cells were collected. The cells were counted using a cell counter (Shanghai Mengwei Biomedical Technology Co., Ltd., SmartCell600A.SC1006) and seeded into 384-well cell culture plates at an appropriate seeding ratio.

[0125] (2) 24 hours after cell seeding, the cells were treated with the drug, designated as day 0. An automated pipette (Thermo, Multidrop 8) was used to add the compounds. The plate was set to 384 wells. The initial concentration of AZD5305 was set to 200 nM, and each well was diluted 1 / 3 with DMSO. Compound 1 was used to set three detection points (0 nM, 20.58 nM, 185.19 nM), with two replicates. The cells were cultured at 37°C in a 5% CO2 incubator.

[0126] (3) On the seventh day, remove the cell culture plate and add an equal volume of CTG reagent (the CTG reagent needs to be pre-mixed according to the reagent instructions to restore the temperature). Gently mix it for 10 minutes (speed 300) on a constant temperature mixer (Hangzhou Ausheng Instrument Co., Ltd., MSC-100). Perform chemiluminescence detection using an ELISA reader (Thermo, Varioskan LUX).

[0127] Data analysis: The CV% of the control group should be less than 30%, and the z' value should be greater than 0.5. Data meeting these quality control results were used to calculate cell viability (inhibition rate (%) = 100 × (control group average - experimental group) / (control group average - culture medium control group average). GraphPad Prism 8 was used to perform nonlinear regression fitting of the inhibition rate curve of the compound and to derive the IC50. 50 value.

[0128] Experimental results: Compound 1, when used in combination with AZD5305, showed a synergistic effect on AZD5305 at both concentrations (see Figure 3).

[0129] Example 4: Clonogenesis inhibition rate test of compound 1 and olaparib in breast cancer cell line HCC1937 (a breast cancer cell line with BRCA1 mutation and Shieldin complex mutation, TNBC).

[0130] Experimental methods: The cell colony formation assay was performed in vitro to investigate the combined effects of compound 1 and olaparib using crystal violet staining. The specific detection methods are as follows:

[0131] (1) Cell culture: The tumor cell line HCC1937 was cultured in an incubator at 37°C and 5% CO2. Cells were passaged at 1 / 3 rate (once every 3 days), and cells in the logarithmic growth phase were used for plating.

[0132] (2) Cell plating: Cells were stained with trypan blue and viable cells were counted. The cell concentration was adjusted to 1000 / well. 2 mL of cell suspension was added to each well of the culture plate. The culture plate was incubated overnight at 37°C, 5% CO2 and 100% relative humidity.

[0133] (3) Compound combination treatment of cells: Take 10 μL of 10 mM compound 1 and add it to 40 μL of DMSO for serial dilution (1 / 5) to obtain working solutions with concentrations of 2 mM, 400 μM, 80 μM, 16 μM, and 3.2 μM; take 10 μL of 20 mM olaparib stock solution and add it to 40 μL of DMSO for serial dilution (1 / 5) to obtain working solutions with concentrations of 4 mM, 800 μM, 160 μM, 32 μM, and 6.4 μM; add 1 μL of the diluted compound working solution to a cell culture plate, and add 2 μL of DMSO to the blank control. Place the six-well cell plate back into the incubator for culture, and replace the culture medium with fresh drug solution as described above.

[0134] (4) Crystal violet staining: Remove the six-well plate from the incubator, remove the culture medium, and wash with PBS. Fix the cells with 1 mL of 100% methanol per well (at room temperature for 15 minutes). After fixation, wash with PBS to remove the methanol. Stain with 1 mL of 0.1% crystal violet per well for 1 hour. After staining, wash three times with 1×PBS until no background staining is visible. Take photos and record the experimental data.

[0135] Data analysis: The cloning area of ​​each well was calculated using ImageJ software. The inhibition rate (IR) of the detected compound was calculated using the following formula: IR (%) = ((blank control – RLU compound) / (blank control)) * 100%. The inhibition rates of different concentrations of the compound were calculated in Excel, and then inhibition curves and related parameters were plotted using GraphPad Prism software.

[0136] Experimental results: Compound 1 itself also has a good inhibitory effect; when compound 1 is used in combination with olaparib, better clonogenic inhibition effect is achieved (see Figures 4 and 5).

[0137] Example 5: Colony formation inhibition rate test of compound 1 and AZD5305 in the pancreatic cancer cell line CAPAN-1 (a pancreatic cancer cell line with a BRCA2 mutation).

[0138] Experimental methods: The cell colony formation effect of the combined use of compound 1 and AZD5305 in vitro was investigated using crystal violet staining. The specific detection methods are as follows:

[0139] (1) Cell culture: The tumor cell line CAPAN-1 was cultured in an incubator at 37°C and 5% CO2. Cells were passaged at 1 / 3 rate (once every 3 days), and cells in the logarithmic growth phase were used for plating.

[0140] (2) Cell plating: Cells were stained with trypan blue and viable cells were counted. The cell concentration was adjusted to 4000 / well. 2 mL of cell suspension was added to each well of the culture plate. The culture plate was incubated overnight at 37°C, 5% CO2 and 100% relative humidity.

[0141] (3) Compound combination treatment of cells: Take 10 μL of 10 mM compound 1 and add it to 40 μL of DMSO for serial dilution (1 / 5) to obtain working solutions with concentrations of 2 mM, 400 μM, 80 μM, 16 μM and 3.2 μM; take 10 μL of compound 1 from 5 mM AZD5305 stock solution and add it to 40 μL of DMSO for serial dilution (1 / 5) to obtain working solutions with concentrations of 1 mM, 200 μM, 40 μM, 8 μM and 1.6 μM; add 1 μL of the diluted compound working solution to a cell culture plate, and add 2 μL of DMSO to the blank control. Place the six-well cell plate back into the incubator for culture, and replace the culture medium with fresh drug solution as described above.

[0142] (4) Crystal violet staining: Remove the six-well plate from the incubator, remove the culture medium, and wash with PBS. Fix the cells with 1 mL of 100% methanol per well (at room temperature for 15 minutes). After fixation, wash with PBS to remove the methanol. Stain with 1 mL of 0.1% crystal violet per well for 1 hour. After staining, wash three times with 1×PBS until no background staining is visible. Take photos and record the experimental data.

[0143] Data Analysis: The clonal area of ​​each well was calculated using ImageJ software. The inhibition rate (IR) of the detected compound was calculated using the following formula: IR (%) = ((blank control – RLU compound) / (blank control)) * 100%. The inhibition rates of different concentrations of the compound were calculated in Excel, and then plotted using the online tool https: / / synergyfinder.fimm.fi. Specifically, the HSA (highest single agent) method was used to calculate the synergistic effect index. When the synergistic effect index < -10, the two drugs have an antagonistic effect; when -10 < synergistic effect index < 10, the two drugs have an additive effect; when the synergistic effect index > 10, the two drugs have a synergistic effect.

[0144] Experimental results: The combination of compound 1 and AZD5305 showed a synergy score of 37.79, indicating a significant synergistic effect between the two drugs (see Table 1, Figure 6 and Figure 7).

[0145] Table 1. Coefficients of colony formation inhibition rates of compound 1 and AZD5305 in the pancreatic cancer cell line CAPAN-1

[0146] Efficacy Test Example 6: Treatment of subcutaneous xenografts of human breast cancer MDA-MB-436 (BRCA1-mutated breast cancer cell line, TNBC) in nude mice.

[0147] Test drug: Compound 2, purity 98.81%. Olaparib, commercially available, purchased from MedChemExpress.

[0148] Preparation method: Compound 2 was dissolved in 5% (volume ratio relative to the final working solution volume; unless otherwise specified, solution preparation steps in the following effect test examples also indicate volume ratios) DMSO, and then prepared into a working solution using 40% PEG400 + 20% (10% TPGS) + 35% H2O. Olaparib was dissolved in a system of DMSO:30% HP-β-CD ((2-hydroxypropyl)-β-cyclodextrin) = 1:9 to obtain the working solution.

[0149] Laboratory animals: BALB / c nude mice, 6-8 weeks old, female. Purchased from Shanghai Jihui Laboratory Animal Breeding Co., Ltd.

[0150] Experimental methods: MDA-MB-436 cells (ATCC, HTB-130) were cultured in vitro as a monolayer under Leibovitz's L-15 medium (containing 10% FBS, 10 μg / ml insulin, and 16 μg / ml glutathione). 0.2 mL of 10 × 10⁻⁶ cells was added...6 One MDA-MB-436 cell (with matrix gel and DPBS at a volume ratio of 1:1) was subcutaneously inoculated into the right posterior dorsal region of each mouse. Twenty-seven days after inoculation, the average tumor volume reached approximately 199.7 mm². 3 Administer medication according to the prescribed regimen. Specific dosages and administration regimens are shown in Table 2. Measure tumor volume and weigh the mice twice weekly, and record the data.

[0151] Table 2. Dosing regimen and evaluation of antitumor efficacy (based on tumor volume calculated on day 35 after administration).

[0152] Note:

[0153] a. Mean ± SEM;

[0154] b. The antitumor efficacy of the compound was evaluated using TGI (%). TGI (%) reflects the tumor growth inhibition rate. The calculation of TGI (%) is as follows: TGI (%) = [1 - (mean tumor volume on day 35 after grouping of a treatment group - mean tumor volume at the start of treatment in that treatment group) / (mean tumor volume on day 35 after grouping of the solvent control group - mean tumor volume at the start of treatment in the solvent control group)] × 100%.

[0155] The cp value was calculated based on tumor volume and analyzed using the Dunnett (2-sided) test (Brown-Forsythe test: p = 0.7489). A p < 0.05 was considered statistically significant.

[0156] The dp value was calculated based on tumor volume and obtained using T-test analysis. A p < 0.05 was considered statistically significant.

[0157] Data statistics: The formula for calculating tumor volume is: V = 0.5a × b 2 , where a and b represent the long and short diameters of the tumor, respectively. The tumor-suppressive efficacy of the compound was evaluated using TGI (%). TGI (%) = [1 - (mean tumor volume at the end of treatment in a certain treatment group - mean tumor volume at the start of treatment in that treatment group) / (mean tumor volume at the end of treatment in the solvent control group - mean tumor volume at the start of treatment in the solvent control group)] × 100%. Data analysis was performed using GraphPad Prism based on data at the end of the trial. T-tests were used for comparisons between two groups. One-way ANOVA was used for comparisons between three or more groups. p < 0.05 was considered statistically significant.

[0158] Experimental Results: In this study, compound 2 showed antitumor activity at 50 mg / kg alone, with a TGI greater than 50%. Compound 2 at 50 mg / kg had a significant synergistic effect with olaparib at 50 mg / kg, achieving tumor regression (TGI greater than 100%, i.e., tumor volume reduced after drug treatment relative to the initial tumor volume) during the administration period. Compared with the olaparib 50 mg / kg monotherapy group, the combination therapy significantly prolonged the tumor regression period after drug withdrawal. The combination regimen was effective and safe, with no mouse deaths or significant weight loss during the entire administration period (see Table 2, Figure 8A and Figure 8B, where data points represent within-group means, error bars represent standard errors (SEM); "*" indicates p < 0.05).

[0159] Efficacy Test Example 7: Treatment of compound 1 and olaparib on NOD-SCID mouse subcutaneous xenografts of human endometrial cancer EN-11-0101 (endometrial cancer with BRCA2 mutation)

[0160] Test drug: Compound 1 p-toluenesulfonate (hereinafter referred to as compound 1 in this example) was prepared as in Example 1, with a free state content of 69.9%. Olaparib, commercially available, was purchased from MedChemExpress.

[0161] Preparation method: Compound 1 was dissolved in 5% DMSO, and then prepared into a working solution using 40% PEG400 + 20% (10% TPGS) + 35% H2O. Olaparib was dissolved in a system of DMSO: 30% HP-β-CD = 1:9 to obtain a working solution.

[0162] Laboratory animals: NOD SCID mice, 6-8 weeks old, female. Purchased from Zhejiang Vital River Laboratory Animal Technology Co., Ltd.

[0163] Experimental Methods: EN-11-0101 is a PDX model of endometrial cancer established by WuXi AppTec. A 30mm... 3 EN-11-0101FP5 tumor tissue blocks were subcutaneously injected into the right posterior back of each mouse, and tumor growth was observed. On day 27 post-inoculation, the average tumor volume reached 122 mmHg. 3 Randomization and administration began at a certain time. Oral administration (PO) was used, with specific dosages and regimens shown in Table 3, where QD indicates once daily. Tumor volume and mouse weight were measured twice weekly, and data were recorded.

[0164] Table 3. Dosing regimen and evaluation of antitumor efficacy (based on tumor volume calculated on day 21 after administration).

[0165] Note:

[0166] a. Mean ± SEM;

[0167] b. The antitumor efficacy of the compound was evaluated using TGI (%). TGI (%) reflects the tumor growth inhibition rate. The calculation of TGI (%) is as follows: TGI (%) = [1 - (mean tumor volume on day 21 after grouping of a treatment group - mean tumor volume at the start of treatment in that treatment group) / (mean tumor volume on day 21 after grouping of the solvent control group - mean tumor volume at the start of treatment in the solvent control group)] × 100%.

[0168] The cp value was calculated based on tumor volume and analyzed using the Dunnett (2-sided) test (Brown-Forsythe test: p = 0.7489). A p < 0.05 was considered statistically significant.

[0169] The dp value was calculated based on tumor volume and obtained using T-test analysis. A p < 0.05 was considered statistically significant.

[0170] Data statistics: The formula for calculating tumor volume is: V = 0.5a × b 2 , where a and b represent the long and short diameters of the tumor, respectively. The tumor-suppressive efficacy of the compound was evaluated using TGI (%). TGI (%) = [1 - (mean tumor volume at the end of treatment in a certain treatment group - mean tumor volume at the start of treatment in that treatment group) / (mean tumor volume at the end of treatment in the solvent control group - mean tumor volume at the start of treatment in the solvent control group)] × 100%. Data analysis was performed using GraphPad Prism based on data at the end of the trial. T-tests were used for comparisons between two groups. One-way ANOVA was used for comparisons between three or more groups. p < 0.05 was considered statistically significant.

[0171] Experimental Results: The EN-11-0101 tumor xenograft model is an olaparib resistance model. In this study, neither olaparib 100 mg / kg nor compound 1 alone showed significant antitumor activity. However, the combination therapy of olaparib 100 mg / kg and compound 1 at a dose of 50 mg / kg showed significantly better antitumor effects than either of the two single-use groups, with a TGI greater than 60%, and a statistically significant difference compared to the solvent control group. The combination regimen was effective and safe, with no mouse deaths or significant weight loss during the entire administration period (see Table 3, Figure 9A and Figure 9B, where data points represent within-group means, error bars represent standard errors (SEM); "*" indicates p < 0.05).

[0172] Efficacy Test Example 8: Treatment of compound 1 and olaparib on NOD-SCID mouse subcutaneous xenografts of human gastric cancer ST-02-0393 (BRCA2-bearing endometrial cancer)

[0173] Test drug: Compound 1 p-toluenesulfonate (hereinafter referred to as compound 1 in this example) was prepared as in Example 1, with a free state content of 69.9%. Olaparib, commercially available, was purchased from MedChemExpress.

[0174] Preparation method: Compound 1 was dissolved in 5% DMSO, and then prepared into a working solution using 40% PEG400 + 20% (10% TPGS) + 35% H2O. Olaparib was dissolved in a system of DMSO: 30% HP-β-CD = 1:9 to obtain a working solution.

[0175] Laboratory animals: NOD SCID mice, 6-8 weeks old, female. Purchased from Zhejiang Vital River Laboratory Animal Technology Co., Ltd.

[0176] Experimental Methods: ST-02-0393 is a gastric cancer PDX model established by WuXi AppTec. A 30mm... 3 ST-02-0393FP6 tumor tissue blocks were subcutaneously injected into the right posterior back of each mouse, and tumor growth was observed. On day 25 post-inoculation, the average tumor volume reached 119 mm. 3 Randomized grouping and drug administration began at that time. Specific dosages and administration regimens are shown in Table 4. Tumor volume was measured twice weekly, and mouse weight was recorded.

[0177] Table 4. Dosing regimen and evaluation of antitumor efficacy (based on tumor volume calculated on day 31 after administration).

[0178] Note:

[0179] a. Mean ± SEM;

[0180] b. The antitumor efficacy of the compound was evaluated using TGI (%). TGI (%) reflects the tumor growth inhibition rate. The calculation of TGI (%) is as follows: TGI (%) = [1 - (mean tumor volume on day 31 after grouping of a treatment group - mean tumor volume at the start of treatment in that treatment group) / (mean tumor volume on day 31 after grouping of the solvent control group - mean tumor volume at the start of treatment in the solvent control group)] × 100%.

[0181] The cp value was calculated based on tumor volume and analyzed using the Dunnett (2-sided) test (Brown-Forsythe test: p = 0.7489). A p < 0.05 was considered statistically significant.

[0182] The dp value was calculated based on tumor volume and obtained using T-test analysis. A p < 0.05 was considered statistically significant.

[0183] Data statistics: The formula for calculating tumor volume is: V = 0.5a × b2 , where a and b represent the long and short diameters of the tumor, respectively. The tumor-suppressive efficacy of the compound was evaluated using TGI (%). TGI (%) = [1 - (mean tumor volume at the end of treatment in a certain treatment group - mean tumor volume at the start of treatment in that treatment group) / (mean tumor volume at the end of treatment in the solvent control group - mean tumor volume at the start of treatment in the solvent control group)] × 100%. Data analysis was performed using GraphPad Prism based on data at the end of the trial. T-tests were used for comparisons between two groups. One-way ANOVA was used for comparisons between three or more groups. p < 0.05 was considered statistically significant.

[0184] Experimental Results: The ST-02-0393 tumor xenograft model is a BRCA2-mutated gastric cancer model. In this study, neither of the two dose groups of compound 1 showed significant antitumor activity when used alone, and olaparib 50 mg / kg showed only partial antitumor activity. However, the combination therapy of olaparib 50 mg / kg and compound 1 50 mg / kg significantly improved the antitumor effect compared to the two single-dose groups, with a TGI greater than 80%, which was statistically significant compared to the olaparib monotherapy group. The combination regimen was effective and safe, with no mouse deaths or significant weight loss during the entire administration period (see Table 4, Figure 10A and Figure 10B, where data points represent within-group means, error bars represent standard errors (SEM); "**" indicates p < 0.01).

[0185] Efficacy test example 9: Compound 1, niraparib, and AZD5305 against human colorectal cancer DLD1 BRCA2 - / - Treatment of subcutaneous xenograft tumors in nude mice

[0186] Test drug: The hydrochloride salt of compound 1 (hereinafter referred to as compound 1 in this example) was prepared as in Example 2, with a free state of 86.2%. Niraparib and AZD5305, commercially available, were purchased from Bidex Pharmaceuticals.

[0187] Preparation method: Compound 1 was dissolved in 5% DMSO, and then prepared into a working solution using 40% PEG400 + 20% (10% TPGS) + 35% H2O. Niraparib was dissolved in a system of 5% DMSO, 40% PEG400, 5% Tween 80 + H2O, and AZD5305 was dissolved in a system of 5% HPMC and 0.1% Tween 80 + H2O to obtain the working solution.

[0188] Laboratory animals: BALB / c nude mice, 6-8 weeks old, female. Purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0189] Experimental method: DLD1 BRCA2 - / -Cells were cultured in a monolayer in vitro under complete culture medium + 10% FBS. 0.1 mL of 5 × 10⁶ cells / mL was added. 6 DLD1 BRCA2 - / - Cells were subcutaneously injected into the right posterior dorsal region of each mouse. Seven days after inoculation, the average tumor volume reached approximately 150 mm². 3 Administer medication according to the prescribed regimen. Specific dosages and administration regimens are shown in Table 5. Measure tumor volume and weigh the mice twice weekly, and record the data.

[0190] Table 5. Dosing regimen and evaluation of antitumor efficacy (all calculated based on tumor volume on day 41 after administration).

[0191] Note:

[0192] a. Mean ± SEM;

[0193] b. The antitumor efficacy of the compound was evaluated using TGI (%). TGI (%) reflects the tumor growth inhibition rate. Calculation of TGI (%): TGI (%) = [1 - (mean tumor volume on day 41 after grouping of a treatment group - mean tumor volume at the start of treatment in that treatment group) / (mean tumor volume on day 41 after grouping of the solvent control group - mean tumor volume at the start of treatment in the solvent control group)] × 100%.

[0194] The cp value was calculated based on tumor volume and analyzed using the Dunnett (2-sided) test (Brown-Forsythe test: p = 0.7489). A p < 0.05 was considered statistically significant.

[0195] The dp value was calculated based on tumor volume and obtained using T-test analysis. A p < 0.05 was considered statistically significant.

[0196] The ep value was calculated based on tumor volume and obtained using T-test analysis. A p < 0.05 was considered statistically significant.

[0197] Data statistics: The formula for calculating tumor volume is: V = 0.5a × b 2 , where a and b represent the long and short diameters of the tumor, respectively. The tumor-suppressive efficacy of the compound was evaluated using TGI (%). TGI (%) = [1 - (mean tumor volume at the end of treatment in a certain treatment group - mean tumor volume at the start of treatment in that treatment group) / (mean tumor volume at the end of treatment in the solvent control group - mean tumor volume at the start of treatment in the solvent control group)] × 100%. Data analysis was performed using GraphPad Prism based on data at the end of the trial. T-tests were used for comparisons between two groups. One-way ANOVA was used for comparisons between three or more groups. p < 0.05 was considered statistically significant.

[0198] Experimental results: DLD1 BRCA2 - / - In a colorectal cancer xenograft model study, compound 1, administered at a dose of 50 mg / kg QD, was combined with niraparib and AZD5305. This combination not only significantly enhanced efficacy during administration but also resulted in sustained tumor regression. While the niraparib and AZD5305 monotherapy groups experienced rapid relapse after drug discontinuation, the combination of compound 1 and both drugs achieved durable tumor regression with no recurrence observed after 90 days. The combination regimen was effective and safe (see Table 5, Figures 11A and 11B. Data points represent within-group means, and error bars represent standard errors (SEM); "**" indicates p < 0.01).

[0199] Efficacy Test Example 10: Compound 1 and low-dose olaparib in human colorectal cancer DLD1 BRCA2 - / - Treatment of subcutaneous xenograft tumors in nude mice

[0200] Test drug: Compound 1, p-toluenesulfonate (hereinafter referred to as compound 1 in this example), was prepared as in Example 1, with a free state content of 69.9%. Olaparib, commercially available, was purchased from Bidex Pharmaceuticals.

[0201] Preparation method: Compound 1 was dissolved in 5% DMSO, and then prepared into a working solution using 40% PEG400 + 20% (10% TPGS) + 35% H2O. Olaparib was dissolved in a system of DMSO: 30% HP-β-CD = 1:9 to obtain a working solution.

[0202] Laboratory animals: BALB / c nude mice, 6-8 weeks old, female. Purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0203] Experimental method: DLD1 BRCA2 - / - Cells were cultured in a monolayer in vitro under complete culture medium + 10% FBS. 0.1 mL of 5 × 10⁶ cells / mL was added. 6 DLD1 BRCA2 - / - Cells were subcutaneously injected into the right posterior dorsal region of each mouse. Seven days after inoculation, the average tumor volume reached approximately 155 mm². 3 Administer medication according to the prescribed regimen. Specific dosages and administration regimens are shown in Table 6. Measure tumor volume and weigh mice twice weekly, and record the data.

[0204] Table 6. Dosing regimen and evaluation of antitumor efficacy (based on tumor volume calculated on day 21 after administration).

[0205] Note:

[0206] a. Mean ± SEM;

[0207] b. The antitumor efficacy of the compound was evaluated using TGI (%). TGI (%) reflects the tumor growth inhibition rate. The calculation of TGI (%) is as follows: TGI (%) = [1 - (mean tumor volume on day 21 after grouping of a treatment group - mean tumor volume at the start of treatment in that treatment group) / (mean tumor volume on day 21 after grouping of the solvent control group - mean tumor volume at the start of treatment in the solvent control group)] × 100%.

[0208] The cp value was calculated based on tumor volume and analyzed using the Dunnett (2-sided) test (Brown-Forsythe test: p = 0.7489). A p < 0.05 was considered statistically significant.

[0209] The dp value was calculated based on tumor volume and obtained using T-test analysis. A p < 0.05 was considered statistically significant.

[0210] Data statistics: The formula for calculating tumor volume is: V = 0.5a × b 2 , where a and b represent the long and short diameters of the tumor, respectively. The tumor-suppressive efficacy of the compound was evaluated using TGI (%). TGI (%) = [1 - (mean tumor volume at the end of treatment in a certain treatment group - mean tumor volume at the start of treatment in that treatment group) / (mean tumor volume at the end of treatment in the solvent control group - mean tumor volume at the start of treatment in the solvent control group)] × 100%. Data analysis was performed using GraphPad Prism based on data at the end of the trial. T-tests were used for comparisons between two groups. One-way ANOVA was used for comparisons between three or more groups. p < 0.05 was considered statistically significant.

[0211] Experimental results: DLD1 BRCA2 - / - In a colorectal cancer xenograft model study, compound 1 significantly enhanced the efficacy of olaparib at an extremely low dose of 0.5 mg / kg QD. Even when the dose of olaparib was reduced to 25 mg / kg QD (below the clinically relevant dose), the combination with compound 1 at 0.25 mg / kg BID still showed excellent synergistic effects, resulting in significant tumor volume reduction. Furthermore, the combination regimen was effective and safe, and also superior to the olaparib monotherapy group in terms of mouse body weight (see Table 6, Figures 12A and 12B. Data points represent within-group means, and error bars represent standard errors (SEM). "**" indicates p < 0.01). These results suggest that compound 1 has the potential to reduce toxicity and enhance efficacy.

[0212] Efficacy Test Example 11: Treatment of compound 1 and olaparib on human breast cancer BR-05-0483E (BRCA1-mutated breast cancer, TNBC) NOD-SCID mouse subcutaneous xenografts.

[0213] Test drug: Compound 1, p-toluenesulfonate, was prepared as in Example 1, with a free compound content of 69.9%. Olaparib, commercially available, purchased from Medchem, lot number 279339.

[0214] Preparation method: Compound 1 was dissolved in 5% DMSO, and then prepared into a working solution using 40% PEG400 + 20% (10% TPGS) + 35% H2O. Olaparib was dissolved in a system of DMSO: 30% HP-β-CD = 1:9 to obtain a working solution.

[0215] Laboratory animals: NOD SCID mice, 6-8 weeks old, female. Purchased from Zhejiang Vital River Laboratory Animal Technology Co., Ltd.

[0216] Experimental Methods: Three days prior to tumor transfer, each mouse was inoculated with a 0.36 mg estrogen tablet (60-day sustained-release) into the left hindquarters. BR-05-0483E is a breast cancer PDX model established by WuXi AppTec. BR-05-0483E cells, after resuscitation and transfer to mice, were defined as FP4. In this experiment, 30 mm... 3 BR-05-0483E FP4 tumor tissue blocks were subcutaneously injected into the right posterior back of each mouse, and tumor growth was observed. On day 21 post-inoculation, the average tumor volume reached 117 mm². 3 Randomized grouping and drug administration began at that time. Specific dosages and administration regimens are shown in Table 7. Tumor volume was measured twice weekly, and mouse weight was recorded.

[0217] Table 7. Dosing regimen and evaluation of antitumor efficacy (based on tumor volume calculated on day 28 after administration).

[0218] Note:

[0219] a. Mean ± SEM;

[0220] b. The antitumor efficacy of the compound was evaluated using TGI (%). TGI (%) reflects the tumor growth inhibition rate. The calculation of TGI (%) is as follows: TGI (%) = [1 - (mean tumor volume on day 28 after grouping of a treatment group - mean tumor volume at the start of treatment in that treatment group) / (mean tumor volume on day 28 after grouping of the solvent control group - mean tumor volume at the start of treatment in the solvent control group)] × 100%.

[0221] The cp value was calculated based on tumor volume and analyzed using the Dunnett (2-sided) test (Brown-Forsythe test: p = 0.7489). A p < 0.05 was considered statistically significant.

[0222] The dp value was calculated based on tumor volume and obtained using T-test analysis. A p < 0.05 was considered statistically significant.

[0223] Data statistics: The formula for calculating tumor volume is: V = 0.5a × b 2 , where a and b represent the long and short diameters of the tumor, respectively. The tumor-suppressive efficacy of the compound was evaluated using TGI (%). TGI (%) = [1 - (mean tumor volume at the end of treatment in a certain treatment group - mean tumor volume at the start of treatment in that treatment group) / (mean tumor volume at the end of treatment in the solvent control group - mean tumor volume at the start of treatment in the solvent control group)] × 100%. Data analysis was performed using GraphPad Prism based on data at the end of the trial. T-tests were used for comparisons between two groups. One-way ANOVA was used for comparisons between three or more groups. p < 0.05 was considered statistically significant.

[0224] Experimental Results: The BR-05-0483E tumor xenograft model is a BRCA1-mutated breast cancer model. In this study, the high and low dose groups of compound 1 showed dose-dependent antitumor activity when used alone. Compared with the olaparib 50 mg / kg monotherapy group, the combination therapy of olaparib 50 mg / kg and compound 1 50 mg / kg showed significantly better antitumor effects than either monotherapy group. After drug withdrawal, the olaparib monotherapy group relapsed quickly, while the combination therapy group remained in a state of tumor regression for about 60 days, significantly delaying relapse. The combination therapy was effective and safe, with no mouse deaths during the entire administration period (see Table 7, Figure 13A and Figure 13B; where data points represent within-group means, error bars represent standard errors (SEM); "**" indicates p < 0.01).

[0225] While specific embodiments of the present invention have been described above, those skilled in the art should understand that these are merely illustrative examples, and various changes or modifications can be made to these embodiments without departing from the principles and essence of the present invention. Therefore, the scope of protection of the present invention is defined by the appended claims.

Claims

1. A combination of a POLθ inhibitor and a PARP inhibitor, characterized in that, The combination includes POLθ inhibitors and PARP inhibitors; the POLθ inhibitors include one or more compounds represented by Formulas 1-3 below and their pharmaceutically acceptable salts:

2. The combination as described in claim 1, characterized in that, The PARP inhibitor is selected from one or more of olaparib, niraparib, tapazoli, pamiparib, rucaparib, fluzoparib, senaparib, saruparib, and their pharmaceutically acceptable salts; and / or, the pharmaceutically acceptable salt of the PARP inhibitor is selected from one or more of hydrochloride, sulfate, trifluoroacetate, formate, and p-toluenesulfonate; and / or, The mass ratio of the POLθ inhibitor to the PARP inhibitor is 1:(0.001-1) or 0.5:(1-300), for example 1:0.002, 1:0.6, 1:1, 1:2, 1:20, 1:50, 1:100 or 1:200; Preferably, the combination comprises one or more of the compounds shown in Formulas 1-3 above, and one or more PARP inhibitors selected from the following: olaparib, niraparib, and Saruparib; More preferably, the combination comprises the compound shown in Formula 3 and olaparib; The combination includes the compound shown in Formula 1 and olaparib; The combination includes the compound shown in Formula 1 and niraparib; The combination comprises the compound shown in Formula 1 and Saruparib; or The combination includes the compound shown in Formula 2 and olaparib.

3. The use of the combination as described in claim 1 or 2 in the preparation of a medicament for treating and / or preventing tumors.

4. The application as described in claim 3, characterized in that, The tumors include one or more of the following: endometrial cancer, lung cancer, breast cancer, gastric cancer, ovarian cancer, prostate cancer, colorectal cancer, pancreatic cancer, uterine leiomyosarcoma, multiple myeloma, cervical cancer, urinary tract-related cancers, skin cancer, and kidney cancer. Preferably, the tumor includes one or more of endometrial cancer, breast cancer, gastric cancer, ovarian cancer, colorectal cancer, and pancreatic cancer.

5. The application as described in claim 3, characterized in that, The tumor includes one or more of the following: tumors deficient in homologous recombination repair genes, tumors deficient in TLS polymerase, tumors deficient in the Shieldin complex, and tumors highly expressing POLθ; preferably, it includes tumors deficient in homologous recombination repair genes and / or tumors deficient in TLS polymerase; more preferably, the tumor meets one or more of the following conditions: (1) The tumors with homologous recombination repair gene deficiency are tumors that are assessed as positive by the HRD assessment system; (2) The homologous recombination repair gene defect is a gene defect that can cause cells to produce HRD, preferably including one or more of the following target genes with decreased expression and / or abnormal protein function: BRCA1, BRCA2, PALB2, RAD51, RAD51C, RAD51B, RAD51D, SLX4, FANCD2, FANCF and XRCC3; preferably, the homologous recombination repair gene defect includes one or more of the following target genes with decreased expression and / or abnormal protein function: BRCA2, BRCA1, XRCC3, RAD51B, RAD51C and FANCF; more preferably, the homologous recombination repair gene defect includes one or more of the following target genes with decreased expression and / or abnormal protein function: BRCA1, BRCA2, RAD51C and XRCC3; (3) The TLS polymerase gene defect includes one or more of the following target genes with decreased expression and / or abnormal protein function: POLI, POLH, RAD18, REV1, REV3, POLK, POLL, PCNA and REV7; preferably, the TLS polymerase defect includes one or more of the following target genes with decreased expression and / or abnormal protein function: REV7, POLE2, POLH and REV3; more preferably, the TLS polymerase gene defect includes one or two of the following target genes with decreased expression and / or abnormal protein function: REV7 and REV3; (4) The Shieldin complex defect includes one or more of the following target genes with decreased expression and / or abnormal protein function: RIF1, 53BP1, SHLD2, SHLD1, SHLD3 and REV7; preferably, the Shieldin complex defect includes one or two of the following target genes with decreased expression and / or abnormal protein function: REV7, RIF1, 53BP1 and SHLD2.

6. The application as described in claim 5, characterized in that, The tumor meets one or more of the following conditions: (1) The tumor includes one or more of the following: endometrial cancer with a deficiency of homologous recombination repair gene, lung cancer with a deficiency of homologous recombination repair gene, breast cancer with a deficiency of homologous recombination repair gene, gastric cancer with a deficiency of homologous recombination repair gene, ovarian cancer with a deficiency of homologous recombination repair gene, prostate cancer with a deficiency of homologous recombination repair gene, colorectal cancer with a deficiency of homologous recombination repair gene, pancreatic cancer with a deficiency of homologous recombination repair gene, uterine leiomyosarcoma with a deficiency of homologous recombination repair gene, multiple myeloma with a deficiency of homologous recombination repair gene, cervical cancer with a deficiency of homologous recombination repair gene, urinary tract-related cancer with a deficiency of homologous recombination repair gene, skin cancer with a deficiency of homologous recombination repair gene, and renal cancer with a deficiency of homologous recombination repair gene; preferably, the tumor further includes a deficiency of Shieldin complex, such as SHLD2 deficiency; (2) The tumors include one or more of the following: TLS polymerase-deficient endometrial cancer, TLS polymerase-deficient lung cancer, TLS polymerase-deficient breast cancer, TLS polymerase-deficient gastric cancer, TLS polymerase-deficient ovarian cancer, TLS polymerase-deficient prostate cancer, TLS polymerase-deficient colorectal cancer, TLS polymerase-deficient pancreatic cancer, TLS polymerase-deficient uterine leiomyosarcoma, TLS polymerase-deficient multiple myeloma, TLS polymerase-deficient cervical cancer, TLS polymerase-deficient urinary tract-associated cancer, TLS polymerase-deficient skin cancer, and TLS polymerase-deficient renal cancer.

7. The application as described in claim 5 or 6, characterized in that, The tumor meets one or more of the following conditions: (1) The homologous recombination repair gene-deficient tumor is homologous recombination repair gene-deficient ovarian cancer, including homologous recombination repair gene-deficient primary ovarian cancer and homologous recombination repair gene-deficient metastatic ovarian cancer. The homologous recombination repair gene-deficient ovarian cancer may include one or more of BRCA2 deficiency, BRCA1 deficiency, XRCC3 deficiency, RAD51B deficiency and RAD51C deficiency; preferably, the homologous recombination repair gene-deficient ovarian cancer further includes one or more of RIF1 deficiency, SHLD2 deficiency and 53BP1 deficiency. (2) The homologous recombination repair gene-deficient tumor is a homologous recombination repair gene-deficient lung cancer, including homologous recombination repair gene-deficient primary lung cancer and homologous recombination repair gene-deficient metastatic lung cancer. The homologous recombination repair gene-deficient lung cancer may include one or more of BRCA2 deficiency, BRCA1 deficiency, XRCC3 deficiency, RAD51B deficiency and RAD51C deficiency; preferably, the homologous recombination repair gene-deficient lung cancer further includes one or more of RIF1 deficiency, SHLD2 deficiency, REV7 deficiency and 53BP1 deficiency. (3) The homologous recombination repair gene-deficient tumor is a homologous recombination repair gene-deficient breast cancer, including homologous recombination repair gene-deficient primary breast cancer and homologous recombination repair gene-deficient metastatic breast cancer. The homologous recombination repair gene-deficient breast cancer may include one or more of BRCA2 deficiency, BRCA1 deficiency, XRCC3 deficiency, RAD51B deficiency and RAD51C deficiency; preferably, the homologous recombination repair gene-deficient breast cancer further includes one or more of RIF1 deficiency, SHLD2 deficiency, REV7 deficiency and 53BP1 deficiency. (4) The homologous recombination repair gene-deficient tumor is homologous recombination repair gene-deficient gastric cancer, including homologous recombination repair gene-deficient primary gastric cancer and homologous recombination repair gene-deficient metastatic gastric cancer. The homologous recombination repair gene-deficient gastric cancer may include one or more of BRCA2 deficiency, BRCA1 deficiency, XRCC3 deficiency, RAD51B deficiency and RAD51C deficiency; preferably, the homologous recombination repair gene-deficient gastric cancer further includes one or more of RIF1 deficiency, SHLD2 deficiency, REV7 deficiency and 53BP1 deficiency. (5) The homologous recombination repair gene-deficient tumor is homologous recombination repair gene-deficient prostate cancer, including homologous recombination repair gene-deficient primary prostate cancer and homologous recombination repair gene-deficient metastatic prostate cancer. The homologous recombination repair gene-deficient prostate cancer may include one or more of BRCA2 deficiency, BRCA1 deficiency, XRCC3 deficiency, RAD51B deficiency and RAD51C deficiency; preferably, the homologous recombination repair gene-deficient prostate cancer further includes one or more of RIF1 deficiency, SHLD2 deficiency, REV7 deficiency and 53BP1 deficiency. (6) The homologous recombination repair gene-deficient tumor is homologous recombination repair gene-deficient colorectal cancer, including homologous recombination repair gene-deficient primary colorectal cancer and homologous recombination repair gene-deficient metastatic colorectal cancer. The homologous recombination repair gene-deficient colorectal cancer may include one or more of BRCA2 deficiency, BRCA1 deficiency, XRCC3 deficiency, RAD51B deficiency, and RAD51C deficiency. Preferably, the homologous recombination repair gene-deficient colorectal cancer further includes one or more of RIF1 deficiency, SHLD2 deficiency, REV7 deficiency, and 53BP1 deficiency colorectal cancer. (7) The homologous recombination repair gene-deficient tumor is homologous recombination repair gene-deficient pancreatic cancer, including homologous recombination repair gene-deficient primary pancreatic cancer and homologous recombination repair gene-deficient metastatic pancreatic cancer. The homologous recombination repair gene-deficient pancreatic cancer may include one or more of BRCA2 deficiency, BRCA1 deficiency, XRCC3 deficiency, RAD51B deficiency and RAD51C deficiency; preferably, the homologous recombination repair gene-deficient pancreatic cancer further includes one or more of RIF1 deficiency, SHLD2 deficiency, REV7 deficiency and 53BP1 deficiency. (8) The homologous recombination repair gene-deficient tumor is a homologous recombination repair gene-deficient uterine leiomyosarcoma, including a homologous recombination repair gene-deficient primary uterine leiomyosarcoma and a homologous recombination repair gene-deficient metastatic uterine leiomyosarcoma. The homologous recombination repair gene-deficient uterine leiomyosarcoma may further include one or more of BRCA2 deficiency, BRCA1 deficiency, XRCC3 deficiency, RAD51B deficiency, and RAD51C deficiency; preferably, the homologous recombination repair gene-deficient uterine leiomyosarcoma further includes one or more of RIF1 deficiency, SHLD2 deficiency, REV7 deficiency, and 53BP1 deficiency. (9) The homologous recombination repair gene-deficient tumor is a homologous recombination repair gene-deficient multiple myeloma, including homologous recombination repair gene-deficient primary multiple myeloma and homologous recombination repair gene-deficient metastatic multiple myeloma. The homologous recombination repair gene-deficient multiple myeloma may further include one or more of BRCA2 deficiency, BRCA1 deficiency, XRCC3 deficiency, RAD51B deficiency, and RAD51C deficiency. Preferably, the homologous recombination repair gene-deficient multiple myeloma further includes one or more of RIF1 deficiency, SHLD2 deficiency, REV7 deficiency, and 53BP1 deficiency. (10) The TLS polymerase-deficient cancers include TLS polymerase-deficient primary cancers and TLS polymerase-deficient metastatic cancers, such as TLS polymerase-deficient primary lung cancer, TLS polymerase-deficient metastatic lung cancer, TLS polymerase-deficient primary breast cancer, TLS polymerase-deficient metastatic breast cancer, TLS polymerase-deficient primary gastric cancer, TLS polymerase-deficient metastatic gastric cancer, TLS polymerase-deficient primary ovarian cancer, TLS polymerase-deficient metastatic ovarian cancer, TLS polymerase-deficient primary prostate cancer, TLS polymerase-deficient metastatic prostate cancer, TLS polymerase-deficient primary colorectal cancer, TLS polymerase-deficient metastatic colorectal cancer, TLS polymerase-deficient primary pancreatic cancer, TLS polymerase-deficient metastatic pancreatic cancer, TLS polymerase-deficient primary uterine leiomyosarcoma, TLS polymerase-deficient metastatic uterine leiomyosarcoma, TLS polymerase-deficient multiple primary myeloma, or TLS polymerase-deficient multiple metastatic myeloma. (11) The homologous recombination repair gene-deficient tumor is a homologous recombination repair gene-deficient endometrial cancer, including homologous recombination repair gene-deficient primary endometrial cancer and homologous recombination repair gene-deficient metastatic endometrial cancer. The homologous recombination repair gene-deficient endometrial cancer may further include one or more of BRCA2 deficiency, BRCA1 deficiency, XRCC3 deficiency, RAD51B deficiency, and RAD51C deficiency. Preferably, the homologous recombination repair gene-deficient endometrial cancer further includes one or more of RIF1 deficiency, SHLD2 deficiency, REV7 deficiency, and 53BP1 deficiency. (13) The Shieldin complex-deficient tumors include one or more of the following: Shieldin complex-deficient endometrial cancer, Shieldin complex-deficient lung cancer, Shieldin complex-deficient breast cancer, Shieldin complex-deficient gastric cancer, Shieldin complex-deficient ovarian cancer, Shieldin complex-deficient prostate cancer, Shieldin complex-deficient colorectal cancer, Shieldin complex-deficient pancreatic cancer, Shieldin complex-deficient uterine leiomyosarcoma, Shieldin complex-deficient multiple myeloma, Shieldin complex-deficient cervical cancer, Shieldin complex-deficient urinary tract-related cancer, Shieldin complex-deficient skin cancer, and Shieldin complex-deficient renal cancer; preferably, the Shieldin complex-deficient tumors include SHLD2-deficient tumors and / or RIF1-deficient tumors; Preferably, the tumor is BRCA2-deficient endometrial cancer, BRCA1-deficient breast cancer, BRCA1-deficient and SHLD2-deficient breast cancer, BRCA2-deficient gastric cancer, RAD51C-deficient and RIF1-deficient gastric cancer, BRCA2-deficient colorectal cancer, or BRCA2-deficient pancreatic cancer.

8. A drug for treating and / or preventing tumors, characterized in that, The drugs include POLθ inhibitors and PARP inhibitors; The POLθ inhibitor and PARP inhibitor are defined as combinations as described in claims 1 and 2, respectively; The tumor is defined as the application as described in any one of claims 4-7; Preferably, the drug satisfies one or more of the following conditions: (1) In the drug, the mass ratio of the POLθ inhibitor to the PARP inhibitor is 1:(0.001-1) or 0.5:(1-300), for example 1:0.002, 1:0.6, 1:1, 1:2, 1:20, 1:50, 1:100 or 1:200; (2) The drug is a gastrointestinal or non-gastrointestinal dosage form, such as capsules, tablets, pills, granules, powders, oral liquids, injections, respiratory dosage forms, mucosal dosage forms, or cavity dosage forms, preferably tablets; (3) The drug is administered by injection or oral administration, such as intravenous injection, subcutaneous injection, peritoneal injection or intramuscular injection; oral administration is preferred.

9. A medicine box set, characterized in that, The pillbox set includes pillbox A and pillbox B, wherein: The cassette A contains a POLθ inhibitor, as defined in the combination described in claim 1; The kit B contains a PARP inhibitor, which is defined as the combination described in claim 2.

10. Application of POLθ inhibitors in the preparation of drugs that enhance the efficacy of PARP inhibitors; The POLθ inhibitor and PARP inhibitor are defined as combinations as described in claims 1 and 2, respectively; Preferably, the PARP inhibitor is effective in inhibiting the function of PARP enzyme; more preferably, the PARP inhibitor is used for the prevention and / or treatment of PARP-related tumors; more preferably, the tumor is defined as described in any one of claims 4-7.

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