Combined use of antibody-drug conjugate and poly(adenosine diphosphate ribose) polymerase inhibitor

Abstract

The present invention relates to the combined use of an antibody-drug conjugate and a poly(adenosine diphosphate ribose) polymerase inhibitor. Specifically, provided is the use of an antibody-drug conjugate or a pharmaceutically acceptable salt, metabolite or solvate thereof individually or combined with a poly(adenosine diphosphate ribose) polymerase inhibitor in the preparation of drugs for preventing and / or treating cancers.

Description

Combined use of antibody-drug conjugates and poly(ADP-ribose) polymerase inhibitors Technical Field

[0001] This application relates to the field of medicine and relates to the use of an antibody-drug conjugate, alone or in combination, in the preparation of a drug for the prevention and / or treatment of cancer. Specifically, the present invention provides an anti-B7 homolog 4 antibody-drug conjugate, or a pharmaceutically acceptable salt, metabolite, or solvate thereof, alone or in combination with a poly(ADP-ribose) polymerase inhibitor, in the preparation of a drug for the prevention and / or treatment of cancer. Background Art

[0002] Antibody-drug conjugates (ADCs) are a class of targeted biologics that link cytotoxic drugs to monoclonal antibodies via a linker. Using the monoclonal antibody as a carrier, small-molecule cytotoxic drugs are efficiently and effectively delivered to target tumor cells in a targeted manner. Tumor-specific antibodies enable ADCs to selectively deliver small-molecule cytotoxic drugs, minimizing off-target effects while preserving their anti-tumor properties, effectively improving the benefit-risk ratio of anti-tumor therapy. B7 homolog 4 is a newly discovered member of the B7 family. It plays a crucial role in multiple cellular biological processes, such as cell differentiation, proliferation, and apoptosis, and may influence tumor cell invasion and metastasis. Furthermore, the B7 family is an important costimulatory molecule that influences processes such as T cell proliferation and B cell activation. Studies have shown that B7 homolog 4 is highly expressed in various tumors, including cholangiocarcinoma, breast cancer, endometrial cancer, non-small cell lung cancer, ovarian cancer, gastric cancer, and pancreatic cancer, while its expression is limited in normal tissues. Therefore, B7 homolog 4 has great potential as a target for ADCs.

[0003] Poly (adenosinediphosphate ribose polymerase, PARP) inhibitors inhibit PARP1-mediated DNA single-strand damage repair, leading to the accumulation of single-strand damage and the initiation of DNA double-strand breaks. In tumors deficient in homologous recombination repair (HRR), DNA double-strand breaks cannot be accurately repaired, resulting in a "synthetic lethality" effect, which limits the anti-tumor activity of PARP inhibitors as single agents.

[0004] Based on the mechanism of action of PARP inhibitors, combining PARP inhibitors with other DNA-damaging drugs can significantly enhance therapeutic efficacy. On the other hand, PARP inhibitors and other DNA-damaging drugs face overlapping toxic side effects, including leukopenia and anemia, which limits the application of related combination therapies. Therefore, combining ADCs loaded with DNA-damaging drugs with PARP inhibitors can reduce drug exposure in normal tissues, improving efficacy while avoiding the accumulation of toxic side effects.

[0005] Summary of the Invention

[0006] The present disclosure provides a use of an antibody-drug conjugate and a poly(ADP-ribose) polymerase inhibitor in combination for preparing a drug for treating cancer, wherein the structure of the antibody-drug conjugate is shown in formula (I):

[0007] n is 1 to 10, preferably 2 to 8, more preferably 3 to 8, and n is a decimal or an integer; Pc is an anti-B7H4 antibody or an antigen-binding fragment thereof.

[0008] In some embodiments, the anti-B7H4 antibodies or antigen-binding fragments thereof described in the present disclosure comprise: heavy chain HCDR1, HCDR2, HCDR3 as shown in the amino acid sequences of SEQ ID NOs: 01, 02, and 03, respectively, and light chain LCDR1, LCDR2, and LCDR3 as shown in the amino acid sequences of SEQ ID NOs: 04, 05, and 06, respectively.

[0009] The CDR sequences mentioned above are shown in the following table:

[0010] Table 1 Heavy chain and light chain CDR sequences

[0011] Note: CDR sequences are derived from those shown in the Kabat definition.

[0012] In some embodiments, the anti-B7H4 antibodies or antigen-binding fragments thereof described in the present disclosure are selected from humanized antibodies or fragments thereof.

[0013] In some embodiments, the anti-B7H4 antibody or antigen-binding fragment thereof described in the present disclosure comprises a heavy chain constant region of human IgG1, IgG2, IgG3 or IgG4 isotype, and a light chain constant region comprising κ or λ; preferably, the anti-B7H4 antibody or antigen-binding fragment thereof comprises a heavy chain constant region of IgG1 or IgG4 isotype.

[0014] In some embodiments, the heavy chain variable region sequence of the anti-B7H4 antibody or antigen-binding fragment thereof described in the present disclosure is as shown in SEQ ID NO: 07 or a variant thereof, and the light chain variable region sequence is as shown in SEQ ID NO: 08 or a variant thereof.

[0015] The sequences of the heavy and light chain variable regions of the aforementioned anti-B7H4 antibodies or antigen-binding fragments thereof are shown below:

[0016] Heavy chain variable region sequence

[0017] Light chain variable region sequence

[0018] Note: The order is FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. The italics in the sequence are FR sequences, and the underlines are CDR sequences. The CDR sequences are derived from the Kabat definition rules.

[0019] In some embodiments, the heavy chain sequence of the anti-B7H4 antibody or antigen-binding fragment thereof described in the present disclosure is as shown in SEQ ID NO: 09 or a variant thereof, and the light chain sequence is as shown in SEQ ID NO: 10 or a variant thereof.

[0020] The sequences of the heavy and light chains of the aforementioned anti-B7H4 antibodies or antigen-binding fragments thereof are shown below:

[0021] Heavy chain (IgG1) amino acid sequence: (SEQ ID NO: 09)

[0022] Light chain (λ) amino acid sequence: (SEQ ID NO: 10)

[0023] In some embodiments, the poly(ADP-ribose) polymerase inhibitor is selected from one or more of Olaparib, Fluzoparib, Niraparib, Pamiparib, Rucaparib, Talazoparib, Veliparib, Senaparib, CEP-8983, BGB-290, or 1'-((7-ethyl-6-carbonyl-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide.

[0024] The combinations described in the present disclosure have additive or synergistic pharmacodynamic effects.

[0025] In an alternative embodiment, the antibody drug conjugate and the poly(ADP-ribose) polymerase inhibitor are contained in different preparations as active ingredients and are administered simultaneously, concurrently, sequentially, continuously, alternately or separately.

[0026] On the other hand, the present application discloses the use of the antibody-drug conjugate and the poly(ADP-ribose) polymerase inhibitor in combination with an anti-VEGF antibody in the preparation of a drug for treating cancer.

[0027] In an optional embodiment, the above-mentioned VEGF antibody is selected from Bevacizumab, Ranibizumab, Sevacizumab, Suvemcitug, Varisacumab, CMAB-801, and LYN-00101.

[0028] In an alternative embodiment, the antibody drug conjugate, the poly(ADP-ribose) polymerase inhibitor, and the anti-VEGF antibody are contained in different preparations as active ingredients, and are administered simultaneously, concurrently, sequentially, continuously, alternately, or separately.

[0029] On the other hand, the above-mentioned antibody-drug conjugate and poly(ADP-ribose) polymerase inhibitor are contained in a single preparation as active ingredients and administered.

[0030] On the other hand, the above-mentioned antibody-drug conjugate, poly(ADP-ribose) polymerase inhibitor, and anti-VEGF antibody are contained in a single preparation as active ingredients and administered.

[0031] In some embodiments, the cancer is a solid tumor. Preferably, the cancer is selected from uterine cancer, breast cancer, ovarian cancer, pancreatic cancer, bile duct cancer, fallopian tube cancer, gastric cancer, colorectal cancer, primary peritoneal cancer, prostate cancer, and lung cancer; more preferably, uterine cancer, breast cancer, and ovarian cancer.

[0032] Further, the cancer is selected from cervical cancer, endometrial cancer, triple-negative breast cancer, ovarian epithelial cancer, epithelial ovarian cancer, platinum-sensitive recurrent ovarian cancer, intrahepatic bile duct cancer, and extrahepatic bile duct cancer; preferably, endometrial cancer, triple-negative breast cancer, and ovarian epithelial cancer; further preferably, the ovarian epithelial cancer is selected from high-grade serous ovarian cancer, low-grade serous ovarian cancer, intrauterine-like ovarian cancer, mucinous ovarian cancer, and ovarian clear cell carcinoma.

[0033] In an optional embodiment, the dosage of the antibody drug conjugate is 0.1 mg / kg to 20.0 mg / kg, preferably 1.0 mg / kg to 15.0 mg / kg; more preferably 1.0 mg / kg, 1.2 mg / kg, 1.4 mg / kg, 1.6 mg / kg, 1.8 mg / kg, 2.0 mg / kg, 2.2 mg / kg, 2.4 mg / kg, 2.6 mg / kg, 2.8 mg / kg, 3.0 mg / kg, 3.2 mg / kg, 3.4 mg / kg, 3.6 mg / kg, 3.8mg / kg, 4.0mg / kg, 4.2mg / kg, 4.4mg / kg, 4.6mg / kg, 4.8mg / kg, 5.0mg / kg, 5.2mg / kg, 5.4mg / kg, 5.6mg / kg, 5.8mg / kg, 6.0mg / kg, 6.2mg / kg, 6.4mg / kg, 6.6mg / kg, 6.8mg / kg, 7.0mg / kg, 7.2mg / kg, 7.4mg / kg, 7.6mg / kg , 7.8mg / kg, 8.0mg / kg, 8.2mg / kg, 8.4mg / kg, 8.6mg / kg, 8.8mg / kg, 9.0mg / kg, 9.2mg / kg, 9.4mg / kg, 9.6mg / kg, 9 .8mg / kg, 10.0mg / kg, 10.2mg / kg, 10.4mg / kg, 10.6mg / kg, 10.8mg / kg, 11.0mg / kg, 11.2mg / kg, 11.4mg / kg, 11.6m g / kg, 11.8mg / kg, 12.0mg / kg, 12.2mg / kg, 12.4mg / kg, 12.6mg / kg, 12.8mg / kg, 13.0mg / kg, 13.2mg / kg, 13.4mg / kg, 13.6mg / kg, 13.8mg / kg, 14.0mg / kg, 14.2mg / kg, 14.4mg / kg, 14.6mg / kg, 14.8mg / kg, 15.0mg / kg or any value in between.

[0034] In alternative embodiments, the antibody drug conjugate is administered once a week, once every two weeks, once every three weeks, or once every four weeks.

[0035] In an optional embodiment, the dosage of the poly(ADP-ribose) polymerase inhibitor is 1-500 mg / kg, preferably 10 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 35 mg / kg, 40 mg / kg, 45 mg / kg, 50 mg / kg, 55 mg / kg, 60 mg / kg, 65 mg / kg, 70 mg / kg, 75 mg / kg, 80 mg / kg, 85 mg / kg, 90 mg / kg, 95 mg / kg, 100 mg / kg, 110 mg / kg, 120 mg / kg, 130 mg / kg, 140 mg / kg, 150 mg / kg, 160 mg / kg. or 500 mg / kg, or any value therebetween.

[0036] In a preferred embodiment, the dosage of the poly(ADP-ribose) polymerase inhibitor is 10-300 mg / kg, more preferably 10 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 35 mg / kg,

[0014] The present invention relates to an oral dosage form of at least one pharmaceutical composition comprising the pharmaceutical composition of the present invention and / or the pharmaceutical composition of the present invention. The pharmaceutical composition of the present invention may be any one of the following: 40 mg / kg, 45 mg / kg, 50 mg / kg, 55 mg / kg, 60 mg / kg, 65 mg / kg, 70 mg / kg, 75 mg / kg, 80 mg / kg, 85 mg / kg, 90 mg / kg, 95 mg / kg, 100 mg / kg, 110 mg / kg, 120 mg / kg, 130 mg / kg, 140 mg / kg, 150 mg / kg, 160 mg / kg, 170 mg / kg, 180 mg / kg, 190 mg / kg, 200 mg / kg, 210 mg / kg, 220 mg / kg, 230 mg / kg, 240 mg / kg, 250 mg / kg, 260 mg / kg, 270 mg / kg, 280 mg / kg, 290 mg / kg, 300 mg / kg, or any number therebetween.

[0037] In alternative embodiments, the poly(ADP-ribose) polymerase inhibitor is administered once a day, twice a day, three times a day, once every two days, or once every three days.

[0038] In an alternative embodiment, the dose of the anti-VEGF antibody is from 1.0 mg / kg to 100 mg / kg, preferably from 1.0 mg / kg to 40 mg / kg, more preferably from 1.0 mg / kg to 30 mg / kg, and further preferably 1.0 mg / kg, 1.2 mg / kg, 1.4 mg / kg, 1.6 mg / kg, 1.8 mg / kg, 2.0 mg / kg, 2.2 mg / kg, 2.4 mg / kg, 2.6 mg / kg, 2.8 mg / kg, 3.0 mg / kg, 3.2 mg / kg, 3.4 mg / kg, 3.6 mg / kg, 3.8 mg / kg, 4.0 mg / kg, 4.2 mg / kg, 4.4 mg / kg, 4.6 mg / kg, 4.8 mg / kg, 5.0 mg / kg, 5.2 mg / kg, 5.4 mg / kg, 5.6 mg / kg, 5.8 mg / kg, 6.0 mg / kg, 6.2 mg / kg, 6.4 mg / kg, 6.6 mg / kg, 6.8 mg / kg, 7.0 mg / kg, 7.2 mg / kg, 7.4 mg / kg, 7.6 mg / kg, 7.8 mg / kg, 8.0 mg / kg, 8.2 mg / kg, 8.4 mg / kg, 8.6 mg / kg, 8.8 mg / kg, 9.0 mg / kg, 9.2 mg / kg, 9.4 mg / kg, 9.6 mg / kg, 9.8 mg / kg, 10.0 mg / kg, 10.2 mg / kg, 10.4 mg / kg, 10.6 mg / kg, 10.8 mg / kg, 11.0 mg / kg, 11.2 mg / kg, 11.4 mg / kg, 11.6 mg / kg, 11.8 mg / kg, 12.0 mg / kg, 12.2 mg / kg, 12.4 mg / kg, 12.6 mg / kg, 12.8 mg / kg, 13.0 mg / kg, 13.2 mg / kg, 13.4 mg / kg, 13.6 mg / kg, 13.8 mg / kg, 14.0 mg / kg, 14.2 mg / kg, 14.4 mg / kg, 14.6 mg / kg, 14.8 mg / kg, 15.0 mg / kg, 15.2 mg / kg, 15.4 mg / kg, 15.6 mg / kg, 15.8 mg / kg, 16.0 mg / kg, 16.2 mg / kg, 16.4 mg / kg, 16.6 mg / kg, 16.8 mg / kg, 17.0 mg / kg, 17.2 mg / kg, 17.4 mg / kg, 17.6 mg / kg, 17.8 mg / kg, 18.0 mg / kg, 18.2 mg / kg, 18.4 mg / kg, 18.6 mg / kg, 18.8 mg / kg, 19.0 mg / kg, 19.2 mg / kg, 19.4 mg / kg, 19.6 mg / kg, 19.8 mg / kg, 20.0mg / kg, 20.2mg / kg, 20.4mg / kg, 20.6mg / kg, 20.8mg / kg, 30.0mg / kg. .

[0039] In alternative embodiments, the anti-VEGF antibody is administered once a week, once every two weeks, or once every three weeks.

[0040] In a preferred embodiment, the dose of the anti-VEGF antibody is 15 mg / kg and the administration frequency is once every three weeks.

[0041] The present disclosure also provides a pharmaceutical composition containing the above-mentioned antibody-drug conjugate and a poly(ADP-ribose) polymerase inhibitor, which comprises one or more pharmaceutically acceptable excipients, diluents or carriers.

[0042] The present disclosure also provides a pharmaceutical composition containing the above-mentioned antibody-drug conjugate, a poly(ADP-ribose) polymerase inhibitor and an anti-VEGF antibody, which contains one or more pharmaceutically acceptable excipients, diluents or carriers.

[0043] In addition, the pharmaceutical composition of the present disclosure can also be administered to patients or subjects in need of such treatment in any suitable manner, such as oral, parenteral, rectal, pulmonary, topical, subcutaneous, intramuscular, or intravenous administration.

[0044] The present disclosure also provides a method for treating cancer, comprising: administering to a patient an effective amount of an antibody-drug conjugate of formula (I) and a poly(ADP-ribose) polymerase inhibitor in combination, wherein the combined administration can be simultaneous, concurrent, sequential, continuous, alternating, or separate administration.

[0045] The present disclosure also provides a method for treating cancer, comprising: administering to a patient an effective amount of an antibody-drug conjugate represented by formula (I), a poly(ADP-ribose) polymerase inhibitor, and an anti-VEGF antibody in combination, wherein the combined administration can be simultaneous, concurrent, sequential, continuous, alternating, or separate.

[0046] The present disclosure also provides a method for treating or preventing cancer, comprising administering the above-mentioned antibody-drug conjugate, an anti-VEGF antibody, and a platinum drug in combination to a subject in need thereof during an induction therapy phase, wherein the combined administration may be simultaneous, concurrent, sequential, continuous, alternating, or separate administration; and administering the above-mentioned antibody-drug conjugate, an anti-VEGF antibody, and a poly(ADP-ribose) polymerase inhibitor in combination to a subject in need thereof during a maintenance therapy phase after the induction therapy, wherein the combined administration may be simultaneous, concurrent, sequential, continuous, alternating, or separate administration.

[0047] In an optional embodiment, the platinum drug is selected from carboplatin, cisplatin, oxaliplatin, nedaplatin, lobaplatin, satraplatin, cycloplatin, miboplatin, enloplatin, iproplatin, dicycloplatin; preferably carboplatin and / or cisplatin.

[0048] In an optional embodiment, the platinum drug is administered at a dose of 10 mg / m 2 Up to 500 mg / m 2 , preferably 10 mg / m 2 Up to 200 mg / m 2 , more preferably 25 mg / m 2 , 50mg / m 2 , 75mg / m 2 , 100mg / m 2 , 125mg / m 2 , 150mg / m 2 , 175mg / m 2 or 200 mg / m 2 The dosing frequency is once a week, once every two weeks, once every three weeks or once every four weeks.

[0049] In an optional embodiment, the dosage of the platinum drug is calculated as the area under the curve (AUC) and is 1 to 20 mg / ml / min, preferably 1 to 10 mg / ml / min, more preferably 2 mg / ml / min, 3 mg / ml / min, 4 mg / ml / min, 5 mg / ml / min, 6 mg / ml / min, 7 mg / ml / min, 8 mg / ml / min, 9 mg / ml / min, and the frequency of administration is once a week, once every two weeks, once every three weeks or once every four weeks.

[0050] In an alternative embodiment, the platinum drug is administered for up to 6 cycles.

[0051] In a preferred embodiment, the dosage of platinum drugs is: cisplatin 75 mg / m 2 Or carboplatin AUC 5mg / ml / min intravenous drip, the dosage frequency is once every three weeks.

[0052] In some embodiments, the cancer is a solid tumor. Preferably, the cancer is selected from uterine cancer, breast cancer, ovarian cancer, pancreatic cancer, bile duct cancer, fallopian tube cancer, gastric cancer, colorectal cancer, primary peritoneal cancer, prostate cancer, and lung cancer; more preferably, uterine cancer, breast cancer, and ovarian cancer.

[0053] In some embodiments, the cancer is selected from cervical cancer, endometrial cancer, triple-negative breast cancer, ovarian epithelial cancer, platinum-sensitive recurrent ovarian cancer, intrahepatic bile duct cancer, and extrahepatic bile duct cancer; preferably, endometrial cancer, triple-negative breast cancer, and ovarian epithelial cancer; further preferably, the ovarian epithelial cancer is selected from high-grade serous ovarian cancer, low-grade serous ovarian cancer, high-grade endometrioid epithelial ovarian cancer, endometrioid ovarian cancer, mucinous ovarian cancer, and ovarian clear cell carcinoma.

[0054] Another aspect of the present disclosure provides the aforementioned anti-B7H4 antibody-drug conjugate for use in treating cancer, wherein the anti-B7H4 antibody-drug conjugate is used in combination with the aforementioned poly(ADP-ribose) polymerase inhibitor.

[0055] Another aspect of the present disclosure provides the aforementioned poly(ADP-ribose) polymerase inhibitor for use in treating cancer, wherein the poly(ADP-ribose) polymerase inhibitor is used in combination with an anti-B7H4 antibody-drug conjugate.

[0056] In the present disclosure, the so-called "combination" is a mode of administration, which includes various situations in which two or more drugs are administered sequentially or simultaneously.

[0057] Administration by simultaneous administration, independent formulation and co-administration, or independent formulation and sequential administration all fall within the scope of combined administration described herein. "Simultaneously" herein refers to administering at least one dose of a poly(ADP-ribose) polymerase inhibitor and an anti-B7H4 antibody-drug conjugate within a certain time period, optionally within 3 days, 2 days, or 1 day, wherein both or more substances exhibit pharmacological effects. "Sequential" administration includes administering a poly(ADP-ribose) polymerase inhibitor and an anti-B7H4 antibody-drug conjugate separately within different dosing cycles. The time period can be within one dosing cycle, optionally within 4 weeks, 3 weeks, 2 weeks, or 1 week. This time period includes treatments in which the poly(ADP-ribose) polymerase inhibitor and the anti-B7H4 antibody-drug conjugate are administered via the same route of administration or different routes of administration.

[0058] the term

[0059] In order to make the present disclosure more easily understood, certain technical and scientific terms are specifically defined below. Unless otherwise explicitly defined herein, all other technical and scientific terms used herein have the meanings commonly understood by those skilled in the art to which the present disclosure belongs.

[0060] The present disclosure incorporates all the contents of application WO2020244657 into the present application.

[0061] The term "antibody drug conjugate" refers to an antibody linked to a biologically active drug via a stable linker. In the present disclosure, "antibody drug conjugate" refers to a monoclonal antibody or antibody fragment linked to a biologically active toxic drug via a stable linker.

[0062] The term "antibody" refers to immunoglobulins, which are tetrapeptide chains composed of two identical heavy chains and two identical light chains connected by interchain disulfide bonds. The amino acid composition and order of the constant region of immunoglobulins' heavy chains vary, resulting in different antigenicity. Consequently, immunoglobulins can be divided into five classes, or isotypes, namely IgM, IgD, IgG, IgA, and IgE, with their corresponding heavy chains being μ, δ, γ, α, and ε, respectively. Within the same class, Igs are further divided into subclasses based on the amino acid composition of their hinge regions and the number and location of heavy chain disulfide bonds. For example, IgG can be divided into IgG1, IgG2, IgG3, and IgG4. Light chains are classified as either kappa or lambda chains based on differences in their constant regions. Each of the five Ig classes can have either kappa or lambda chains.

[0063] The approximately 110 amino acids near the N-terminus of an antibody's heavy and light chains vary greatly in sequence, forming the variable region (Fv region); the remaining amino acid sequences near the C-terminus are relatively stable, forming the constant region. The variable region comprises three hypervariable regions (HVRs) and four framework regions (FRs), whose sequences are relatively conserved. These three hypervariable regions determine the antibody's specificity and are also known as complementarity-determining regions (CDRs). Each light chain variable region (LCVR) and heavy chain variable region (HCVR) consists of three CDR regions and four FR regions, arranged in the following order from amino to carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The three CDR regions of the light chain are LCDR1, LCDR2, and LCDR3; the three CDR regions of the heavy chain are HCDR1, HCDR2, and HCDR3.

[0064] In the present disclosure, the amino acid sequences of the above CDRs are shown according to the Kabat definition rules. However, it is well known to those skilled in the art that antibody CDRs can be defined in the art by a variety of methods, such as Chothia based on the three-dimensional structure of the antibody and the topology of the CDR loop (Chothia et al. (1989) Nature 342: 877-883, Al-Lazikani et al., "Standard conformations for the canonical structures of immunoglobulins", Journal of Molecular Biology, 273, 927-948 (1997)), Kabat based on antibody sequence variability (Kabat et al., Sequences of Proteins of Immunological Interest, 4th Edition, US Department of Health and Human Services, National Institutes of Health (1987)), AbM (University of Bath), Contact (University College London), the international ImMunoGeneTics database (IMGT) (world wide web imgt.cines.fr / ), and the North CDR definition based on affinity propagation clustering using a large number of crystal structures. It will be understood by those skilled in the art that, unless otherwise specified, the terms "CDR" and "complementarity determining region" of a given antibody or region thereof (e.g., variable region) should be understood to encompass complementarity determining regions defined by any of the above-mentioned known schemes described herein. Although the scope of protection claimed in the present invention is based on the sequences shown in the Kabat definition rules, amino acid sequences corresponding to other CDR definition rules should also fall within the scope of protection of the present invention.

[0065] The term "antigen-binding fragment" refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen. It has been shown that fragments of a full-length antibody can be used to perform the antigen-binding function of an antibody. Examples of binding fragments included in "antigen-binding fragments" include (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL, and CH1 domains; (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments connected by a disulfide bridge on the hinge region; (iii) a Fd fragment consisting of the VH and CH1 domains; (iv) a Fv fragment consisting of the VH and VL domains of a single arm of an antibody; (v) a single domain or dAb fragment (Ward et al., (1989) Nature 341: 544-546), which consists of a VH domain; and (vi) isolated complementarity determining regions (CDRs) or (vii) a combination of two or more isolated CDRs, optionally connected by a synthetic linker.

[0066] The term "drug loading" refers to the average number of cytotoxic drugs loaded per ligand in a molecule of Formula (I), and can also be expressed as the ratio of the amount of drug to the amount of antibody. The drug loading can range from 0 to 12, preferably 1 to 10, cytotoxic drugs (D) attached per antibody (Pc). In the embodiments of the present disclosure, the drug loading is expressed as n, also known as the DAR value, and exemplary values ​​are 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. The average number of drug products per ADC molecule after the conjugation reaction can be determined by conventional methods such as UV / visible spectroscopy, mass spectrometry, ELISA assays, and HPLC characterization.

[0067] The term "pharmaceutical composition" is a product comprising one or more active ingredients (e.g., antibodies, ADCs) in optionally specified amounts, as well as any product produced directly or indirectly by combining one or more active ingredients in optionally specified amounts. The different active ingredients in the pharmaceutical composition can be administered independently in separate formulations, including administration simultaneously or at different time points for combined synergistic effect. In the present disclosure, "pharmaceutical composition" and "formulation" are not mutually exclusive.

[0068] The term "treating" means administering an internal or external therapeutic agent, such as a composition comprising any of the binding compounds of the present disclosure, to a patient who has one or more symptoms of a disease for which the therapeutic agent is known to have a therapeutic effect. Typically, the therapeutic agent is administered in an amount effective to alleviate one or more symptoms of the disease in the treated patient or population to induce regression of such symptoms or inhibit the development of such symptoms to any clinically measurable degree. The amount of a therapeutic agent effective to alleviate any specific disease symptom (also referred to as a "therapeutically effective amount") can vary according to a variety of factors, such as the patient's disease state, age, and weight, and the ability of the drug to produce the desired therapeutic effect in the patient. Whether the symptoms of the disease have been alleviated can be assessed by any clinical test method commonly used by a physician or other health care professional to assess the severity or progression of the symptoms. Although embodiments of the present disclosure (e.g., methods of treatment or articles of manufacture) may not be effective in alleviating every symptom of the target disease, they should alleviate the target disease symptoms in a statistically significant number of patients as determined by any statistical test known in the art, such as Student's t-test, chi-square test, U test according to Mann and Whitney, Kruskal-Wallis test (H test), Jonckheere-Terpstra test, and Wilcoxon test. BRIEF DESCRIPTION OF THE DRAWINGS

[0069] Figure 1. Combined index of drug A combined with drug B in inhibiting proliferation of 7 ovarian cancer cell lines.

[0070] Figure 2 Combination index of drug A combined with drug B in inhibiting proliferation of 4 breast cancer cell lines.

[0071] Figure 3. Combined index of drug A combined with drug B in inhibiting the proliferation of 8 endometrial cancer cell lines.

[0072] Figure 4 shows the efficacy of drug A combined with drug B on the human breast cancer cell MX-1 subcutaneous xenograft tumor model.

[0073] Figure 5 Effects of drug A combined with drug B on the body weight of mice bearing human breast cancer cells MX-1.

[0074] Figure 6 shows the efficacy of drug A combined with drug B on the human ovarian cancer PDX model OV0243 subcutaneous xenograft tumor model.

[0075] Figure 7 Effects of drug A combined with drug B on the body weight of OV0243 tumor-bearing mice in the human ovarian cancer PDX model.

[0076] Figure 8 shows the efficacy of drug A combined with drug B on the human endometrial cancer cell RL95-2 subcutaneous xenograft tumor model.

[0077] Figure 9 Effects of drug A combined with drug B on the body weight of mice bearing human endometrial cancer cells RL95-2. DETAILED DESCRIPTION

[0078] The present application will be explained in more detail below with reference to the embodiments. The embodiments of the present application are only used to illustrate the technical solutions of the present application and are not intended to limit the essence and scope of the present application.

[0079] Example 1. Preparation of anti-B7H4 antibody drug conjugates and PARP inhibitors

[0080] According to the production methods described in WO2020244657, hu2F7 (an anti-B7H4 antibody) and an isotecan analog were used to prepare an anti-B7H4 antibody-drug conjugate as shown in the following structure. The HIC method calculated the average value: y = 6.1. The hu2F7 heavy chain sequence is shown in SEQ ID NO: 09, and the light chain sequence is shown in SEQ ID NO: 10. The anti-B7H4 antibody-drug conjugate serves as Drug A.

[0081] 1'-((7-ethyl-6-carbonyl-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide was prepared by the method disclosed in WO2022223025 and used as drug B.

[0082] Example 2. Evaluation of the inhibitory effect of drug A in combination with drug B on the proliferation of human ovarian cancer cells, human breast cancer cells and human endometrial cancer cells.

[0083] 1. Experimental Materials

[0084] 1.1 Cell lines and experimental reagents

[0085] Human ovarian cancer cells OAW42, KURAMOCHI, CAOV3, COV362, and OAW28; human breast cancer cells HCC38, MX-1, CAL-148, and CAL-51; and human endometrial cancer cells EN, RL95-2, Ishilawa, EFE184, MFE280, and KLE were purchased from Nanjing Kebai. Human ovarian cancer cells OVCAR-3 and human endometrial cancer cells AN3CA were purchased from ATCC, and human ovarian cancer cells SK-OV-3 were purchased from the Chinese Academy of Sciences Cell Bank. Human endometrial cancer cells HEC-1A were purchased from Guangzhou Geneo.

[0086] Drug A was prepared and dissolved in PBS, and drug B was prepared and dissolved in DMSO;

[0087] Luminescent Cell Viability Assay was purchased from Promega, catalog number G7573;

[0088] 1640 culture medium was purchased from Gibco, catalog number 22400-089;

[0089] Mccoy 5a medium was purchased from Gibco, catalog number 16600-082;

[0090] DMEM medium was purchased from Gibco, catalog number 11995-065;

[0091] EMEM medium was purchased from ATCC, catalog number 30-2003;

[0092] FBS was purchased from Gibco, catalog number 10091-148;

[0093] Pancreatin was purchased from Gibco, catalog number 25200-072;

[0094] PBS was purchased from Gibco with the product number 10010-023.

[0095] 1.2 Instruments:

[0096] Microplate reader (BioTek Synergy H1); pipette (Eppendorf).

[0097] 2. Experimental methods and processing results

[0098] 2.1 Evaluation of the combined effects of drug A and drug B on the growth inhibition of human ovarian cancer, breast cancer, and endometrial cancer cells

[0099] Tumor cells in good growth condition were inoculated into 96-well plates. After overnight adherence and growth, different concentrations of test substances were administered, and the two test substances were cross-acted at different concentrations.

[0100] The starting concentration of test drug A was set at 1000 nM, with a 1:3 dilution, and a total of six concentration gradients. The starting concentration of test drug B was set at 1000 nM, with a 1:3 dilution, and a total of six concentration gradients. The specific drug action schedule is shown in Table 2-1. Six days after the test substances were applied to each cell type, the CTG method was used to detect the in vitro proliferation inhibitory effect of the test drug combination on tumor cells.

[0101] Table 2-1 Dosage regimen of drug A and drug B

[0102] According to the signal value measured by the microplate reader, the cell growth inhibition rate was calculated according to the following formula:

[0103] Growth inhibition rate (%) = [1-(measured value - minimum mean value - background value) / (maximum mean value - background value - minimum mean value)] × 100% (measured value: compound well reading; minimum value: ZPE well reading (well reading without cell culture medium); maximum value: HPE well reading (DMSO-treated cell reading); background value: cell background value before compound addition).

[0104] The results of the combined effects of Drug A and Drug B on cell growth inhibition are shown in Tables 2-2 through 2-20. Drugs A and B inhibited tumor cell growth in a concentration-dependent manner across different concentration ranges. The combined effect of the two drugs significantly enhanced cell growth inhibition, demonstrating additive or synergistic effects.

[0105] Table 2-2 Inhibitory effect of drug A and drug B on the growth of OVCAR-3 cells

[0106] Table 2-3 Inhibitory effect of drug A and drug B on SK-OV-3 cell growth

[0107] Table 2-4 Inhibitory effect of drug A and drug B on the growth of OAW42 cells

[0108] Table 2-5 Inhibitory effect of drug A and drug B on KURAMOCHI cell growth

[0109] Table 2-6 Inhibitory effect of drug A and drug B on CAOV3 cell growth

[0110] Table 2-7 Inhibitory effect of drug A and drug B on COV362 cell growth

[0111] Table 2-8 Inhibitory effect of drug A and drug B on the growth of OAW28 cells

[0112] Table 2-9 Inhibitory effect of drug A and drug B on HCC38 cell growth

[0113] Table 2-10 Inhibitory effect of drug A and drug B on MX-1 cell growth

[0114] Table 2-11 Inhibitory effect of drug A and drug B combined on CAL-148 cell growth

[0115] Table 2-12 Inhibitory effect of the combination of drug A and drug B on the growth of CAL-51 cells

[0116] Table 2-13 Inhibitory effect of the combination of drug A and drug B on the growth of RL95-2 cells

[0117] Table 2-14 Inhibitory effect of the combination of drug A and drug B on the growth of HEC-1A cells

[0118] Table 2-15 Inhibitory effect of the combination of drug A and drug B on the growth of EN cells

[0119] Table 2-16 Inhibitory effect of the combination of drug A and drug B on the growth of Ishikawa cells

[0120] Table 2-17 Inhibitory effect of the combination of drug A and drug B on the growth of AN3CA cells

[0121] Table 2-18 Inhibitory effect of the combination of drug A and drug B on the growth of EFE184 cells

[0122] Table 2-19 Inhibitory effect of the combination of drug A and drug B on the growth of MFE280 cells

[0123] Table 2-20 Inhibitory effect of the combination of drug A and drug B on the growth of KLE cells

[0124] 2.2 Evaluation of the combined inhibitory effect of drug A combined with drug B on the growth of tumor cells

[0125] Based on the experimental results of Experiment 2.1, the cell growth inhibition rate was converted into cell survival rate, and the data of the tumor cell growth inhibition experiment were analyzed using Combenefit software (Di Veroli et al., 2016, Bioinformatics 32(18):2866-2868). The Bliss model was used to evaluate the synergistic / antagonistic effect, and the combined index (CI) of the two drugs was calculated. Among them, CI>10 indicates a synergistic effect of drug efficacy, -10<CI<10 indicates an additive effect of drug efficacy, and CI<-10 indicates an antagonistic effect of drug efficacy.

[0126] Figures 1, 2, and 3 show that the combination index of the two drugs was CI>10 in most cell lines, indicating that the two drugs had a synergistic effect in most cell lines. In addition, under different concentration combinations of the two drugs, the vast majority of CIs were >-10 or CI>10, indicating that the different concentration combinations of the two drugs had an additive or synergistic effect.

[0127] 3. Experimental conclusions:

[0128] The combination of drug A and drug B can enhance the inhibitory effect on tumor cell growth compared with single drug. The combination of drug A and drug B has a synergistic and additive effect.

[0129] Example 3. Evaluation of the Inhibitory Effect of Drug A and Drug B Combined on MX-1 Human Breast Cancer Cell Transplanted Tumors in Mice

[0130] 1. Experimental Materials

[0131] Drug A was prepared by diluting with normal saline, and drug B was prepared by diluting with 0.5% HPMC;

[0132] Human breast cancer MX-1 cells were cultured as monolayers in DMEM / F12 medium supplemented with 10% fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin at 37°C in a 5% CO2 incubator. Cells were routinely digested and passaged twice weekly using trypsin-EDTA. When cell saturation reached 80%-90% and the desired number of cells was reached, cells were harvested, counted, and plated.

[0133] Balb / c nude female mice, weighing 17-25 g, were purchased from Shanghai Bikeway Biotechnology Co., Ltd.

[0134] 2. Experimental Methods

[0135] MX-1 cells were resuspended in PBS at a density of 100 × 10 6 / mL, the resuspended cells were mixed with an equal volume of Matrigel and inoculated subcutaneously on the right back of each mouse, with 0.1 mL (5×10 6 cells / animal), and wait until the average tumor volume grows to 100-150mm 3 Grouping was performed at day 1 (D1), and dosing began on the same day. Mice were administered orally (po) once daily, and mice were administered via tail vein injection (iv) as a single dose. The dosing volume was 10 mL / kg. The solvent group received the same volume of solvent. Specific dosing schedules are shown in Table 3-1. Tumor volume was measured, mice were weighed, and the data were recorded.

[0136] The experimental indicator is to examine the effect of drugs on tumor growth, and the specific indicator is tumor inhibition rate TGI (%).

[0137] The tumor diameter was measured with a vernier caliper twice a week, and the tumor volume (V) was calculated as follows: V = 1 / 2 × a × b 2 Where a and b represent length and width respectively.

[0138] Calculation of TGI (%): If the tumor did not regress, TGI (%) = [1 - (average tumor volume at the end of dosing for a given treatment group - average tumor volume at the time of grouping for that treatment group) / (average tumor volume at the end of treatment for the vehicle control group - average tumor volume at the time of grouping for the vehicle control group)] × 100%. If the tumor regressed, TGI (%) = [1 - (average tumor volume at the end of dosing for a given treatment group - average tumor volume at the time of grouping for that treatment group) / average tumor volume at the time of grouping for that treatment group] × 100%.

[0139] The experiment ends, the experimental endpoint is reached, or the average tumor volume reaches 2000mm 3 The animals were killed by CO2 anesthesia, and then the tumors were removed by dissection and photographed.

[0140] Experimental data were analyzed and graphed using GraphPad Prism 9.4. Tumor volume data for each group at different time points were statistically analyzed using Dunnett's multiple comparisons test in a two-way ANOVA to assess intergroup differences. Differences in tumor volume between the two groups were analyzed using the t-test, with a p < 0.05 defining the difference as statistically significant.

[0141] Table 3-1. Dosing regimen of drug A combined with drug B in the MX-1 model

[0142] 3. Experimental Results

[0143] The growth inhibitory effects of Drug A combined with Drug B on the MX-1 model are shown in Table 3-2 and Figure 4 , and the changes in body weight of animals in each group of the MX-1 model are shown in Figure 5 . During the experimental period, the average body weight of animals in each group remained relatively stable. No mice experienced treatment-related deaths or other abnormal symptoms, indicating that tumor-bearing mice were well tolerated by the test drugs, including monotherapy with Drug A, monotherapy with Drug B, and the combination of Drugs A and B.

[0144] Table 3-2. Growth inhibitory effects of drug A combined with drug B on the MX-1 model

[0145] p-value D28: Values ​​calculated by Dunnett analysis using two-way ANOVA based on tumor volume in each group, with the vehicle group as the control. Statistical analysis was also performed using the t-test. The p-values ​​for the 0.3 mg / kg plus 1 mg / kg group, compared with the 0.3 mg / kg and 1 mg / kg groups, were <0.0001 and 0.0015, respectively. The p-values ​​for the 0.3 mg / kg plus 2 mg / kg group, compared with the 0.3 mg / kg and 2 mg / kg groups, were <0.0001 and 0.2139, respectively.

[0146] 4. Experimental conclusions:

[0147] In the MX-1 human breast cancer transplant mouse model, the tumor-bearing mice were tolerant to drug A monotherapy or combined drug B treatment. The anti-tumor efficacy of the drug B_0.3mg / kg and drug A_1mg / kg combination treatment group was significantly better than that of each single drug treatment group. The anti-tumor efficacy of the drug B_0.3mg / kg and drug A_2mg / kg combination treatment group was significantly better than that of the drug B_0.3mg / kg monotherapy group, and slightly better than that of the drug A_2mg / kg monotherapy group.

[0148] Example 4. Evaluation of the in vivo inhibitory effect of drug A combined with drug B on human ovarian cancer OV0243 in a mouse subcutaneous xenograft tumor model

[0149] 1. Experimental Materials

[0150] Drug A was prepared by diluting with normal saline, and drug B was prepared by diluting with 0.5% HPMC;

[0151] OV0243 (mild cachexia), human ovarian cancer tumor tissue, was provided by Crown Biotech (Beijing) Co., Ltd.

[0152] BALB / c nude female mice, weighing 16-22 g, were purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd.

[0153] 2. Experimental Methods

[0154] OV0243 human ovarian cancer tissue was evenly cut into tumor masses with a diameter of 2-3 mm and inoculated subcutaneously at the right anterior scapula of BALB / cnude mice. The tumor growth was observed regularly. When the average tumor volume reached 100-150 mm 3 The mice were randomly divided into groups according to the tumor size and weight as shown in Table 4-1. The tumor diameter was measured with a vernier caliper twice a week, and the tumor volume (V) was calculated as follows: V = 1 / 2 × a × b 2 Where a and b represent length and width respectively.

[0155] The experimental indicator is to examine the effect of drugs on tumor growth, and the specific indicator is tumor inhibition rate TGI (%).

[0156] Calculation of TGI (%): If the tumor did not regress, TGI (%) = [1 - (average tumor volume at the end of dosing for a given treatment group - average tumor volume at the time of grouping for that treatment group) / (average tumor volume at the end of treatment for the vehicle control group - average tumor volume at the time of grouping for the vehicle control group)] × 100%. If the tumor regressed, TGI (%) = [1 - (average tumor volume at the end of dosing for a given treatment group - average tumor volume at the time of grouping for that treatment group) / average tumor volume at the time of grouping for that treatment group] × 100%.

[0157] The experiment ends, the experimental endpoint is reached, or the average tumor volume reaches 2000mm 3 The animals were killed by CO2 anesthesia, and then the tumors were removed by dissection and photographed.

[0158] Experimental data were analyzed and graphed using GraphPad Prism 9.4. Tumor volume data for each group at different time points were statistically analyzed using Dunnett's multiple comparisons test in a two-way ANOVA to assess intergroup differences. Differences in tumor volume between the two groups were analyzed using the t-test, with a p < 0.05 defining the difference as statistically significant.

[0159] The invivoSyn method was used to analyze the synergistic effect of drug combination therapy. First, the tumor growth rate (eGR, exponential growth rate) of each mouse and the average tumor growth rate (eGRg) of each group were calculated based on the exponential growth model. Then, the conditional probability formula Pr(T|C)=S was used to calculate the synergistic effect of drug combination therapy. T / S C The relative survival rates of the drug-treated group and the control group were calculated, where S A 、S B and S AB They are defined as the relative survival rates of single-drug group A, single-drug group B and combination drug group at time point t, respectively.

[0160] The expected survival rate E(S) of the combined drug group was calculated based on the single drug highest effect model (HSA, Highest Single Agent) AB ). For the HSA model, E(S AB )=min(S A ,S B CI (Combination Index) is defined as the survival rate S observed in the combined drug group AB With E(SAB ) ratio, and the synergy score (SS, Synergy Score) is defined as E(S AB ) and S AB The percentage difference between the two groups was 0. A synergistic effect was defined when the CI value was less than 1 or the SS value was greater than 0. The p-value and confidence interval were calculated using the stratified bootstrap method.

[0161] Table 4-1. Dosing regimen of drug A combined with drug B in the OV0243 model

[0162] 3. Experimental Results

[0163] The growth inhibitory effects of drug A combined with drug B on the OV0243 model are shown in Table 4-2 and Figure 6 , and the synergistic effect analysis of drug A combined with drug B is shown in Table 4-3 . Body weight changes of animals in each group in the OV0243 model are shown in Figure 7 .

[0164] On day 27 after dosing, 5 animals (5 / 6) in the 0.1 mg / kg drug B group showed partial tumor regression, 3 animals (3 / 6) in the 1 mg / kg drug B group showed partial tumor regression, and 3 animals (3 / 6) showed complete tumor regression. Five animals (5 / 6) in the 5 mg / kg drug A group showed complete tumor regression, and one animal (1 / 6) showed partial tumor regression. All animals in the 0.1 mg / kg drug B plus 5 mg / kg drug A groups and the 1 mg / kg drug B plus 5 mg / kg drug A groups showed complete tumor regression.

[0165] Based on the data statistics from D0-D27, the average tumor volume of each treatment group decreased to varying degrees compared with the solvent group, and all showed statistically significant differences (p < 0.0001). In addition, based on the comparison of tumor volume between the treatment groups on day 27 after administration, the tumor inhibition effect of the drug B_0.1mg / kg combined with drug A_5mg / kg treatment group was significantly better than the drug B_0.1mg / kg treatment group (p < 0.0001) and slightly better than the drug A_5mg / kg treatment group (p = 0.3409). The drug B_1mg / kg combined with drug A_5mg / kg treatment group was slightly better than the drug B_1mg / kg treatment group and the drug A_5mg / kg treatment group (p = 0.0950, p = 0.3409).

[0166] Based on tumor volume on day 83 after dosing (i.e., day 56 after drug withdrawal), statistics showed that the group treated with 0.1 mg / kg of drug B plus 5 mg / kg of drug A had significantly better tumor inhibition than the group treated with 0.1 mg / kg of drug B (p=0.0101) and slightly better than the group treated with 5 mg / kg of drug A (p=0.3409). The group treated with 1 mg / kg of drug B plus 5 mg / kg of drug A had slightly better tumor inhibition than the group treated with 1 mg / kg of drug B and the group treated with 5 mg / kg of drug A (p=0.0603, p=0.3409, respectively).

[0167] When drug A 5 mg / kg was combined with drug B 0.1 mg / kg and drug B 1 mg / kg, the HSA model-based CI values ​​were all less than 1 (0.724 and 0.741, respectively), the Synergy Score (SS) was greater than 0 (0.503 and 0.471, respectively), and the p-values ​​were all less than 0.05. Therefore, the combination therapy had a significant synergistic effect, and the efficacy of the combination group was significantly higher than that of the single drug.

[0168] During the experiment, the average body weight of animals in the vehicle group was well maintained, with an average body weight change of 5.70% on day 27. The average body weight changes in the drug B 0.1 mg / kg, drug B 1 mg / kg, drug A 5 mg / kg, drug B 0.1 mg / kg combined with drug A 5 mg / kg, and drug B 1 mg / kg combined with drug A 5 mg / kg groups were 2.18%, 3.75%, 4.59%, 2.61%, and 2.85%, respectively. The OV0243 model exhibits mild cachexia. During dosing, one animal in the drug B 0.1 mg / kg group lost 17.39% of its body weight on day 23. Despite discontinuation of dosing (Days 23-27) and supplementation with nutritional gel, the mouse's weight did not recover. By day 51, the weight loss exceeded 20%, and the animal was euthanized in accordance with animal ethics regulations (individual differences in drug tolerance or underlying medical conditions cannot be ruled out). In addition, no other animals discontinued the drug due to weight loss during the entire experiment, and no animals became ill or died, indicating that tumor-bearing mice were able to tolerate the test drug A monotherapy, drug B monotherapy, and the combination of drugs A and B well.

[0169] Table 4-2. Growth inhibitory effect of drug A combined with drug B on the OV0243 model

[0170] p-value D27: Value obtained by Dunnett analysis using two-way ANOVA based on tumor volume in each group, with the vehicle group as the control. Additionally, based on tumor volume on day 27 after administration, statistical analysis was performed using a t-test. The p-values ​​for the 0.1 mg / kg plus 5 mg / kg group compared to the 0.1 mg / kg and 5 mg / kg groups were <0.0001 and 0.3409, respectively. The p-values ​​for the 1 mg / kg plus 5 mg / kg group compared to the 1 mg / kg and 5 mg / kg groups were 0.0950 and 0.3409, respectively, indicating no statistically significant differences. Statistical analysis was performed using a t-test based on tumor volume on day 83 after administration. The p-values ​​for the 0.1 mg / kg drug B plus 5 mg / kg drug group compared to the 0.1 mg / kg drug B group and the 5 mg / kg drug A group were 0.0101 and 0.3409, respectively. The p-values ​​for the 1 mg / kg drug B plus 5 mg / kg drug group compared to the 1 mg / kg drug B group and the 5 mg / kg drug A group were 0.0603 and 0.3409, respectively.

[0171] Table 4-3. Analysis of synergistic effects of drug A alone and in combination with drug B based on the HSA model

[0172] 4. Experimental conclusions:

[0173] In the OV0243 human ovarian cancer xenograft mouse model, tumor-bearing mice tolerated both drug A monotherapy, drug B monotherapy, and the combination of drugs A and B well. Both drug A and drug B monotherapy significantly inhibited the growth of xenograft tumors in nude mice. Drug A combined with drug B at 0.1 mg / kg and drug A combined with drug B at 1 mg / kg also demonstrated significant tumor inhibition, slightly superior to either drug A or drug B monotherapy. The combination therapy groups demonstrated a significant synergistic effect.

[0174] Example 5. Evaluation of the Inhibitory Effect of Drug A and Drug B Combined on RL95-2 Endometrial Cancer Cell Transplanted Tumors in Mice

[0175] 1. Experimental Materials

[0176] Drug A was prepared by diluting with normal saline, and drug B was prepared by diluting with 0.5% HPMC;

[0177] Human endometrial cancer cells RL95-2 were cultured in a 37°C, 5% CO2 incubator. Twice weekly, cells were routinely digested and passaged using trypsin-EDTA. When cell saturation reached 80%-90% and the desired number of cells was reached, cells were harvested, counted, and plated.

[0178] 2. Experimental Methods

[0179] RL95-2 cells were resuspended in PBS at a density of 100 × 10 6 / mL, the resuspended cells were mixed with an equal volume of Matrigel and inoculated subcutaneously on the right back of each mouse, with 0.1 mL (5×10 6 cells / animal), and wait until the average tumor volume grows to 100-150mm 3 Groups were assigned at day 1 (D1), and dosing began on the same day. Mice were administered orally (po) once daily; mice were administered via tail vein injection (iv) as a single dose; the dosing volume was 10 mL / kg. The solvent group received the same volume of solvent. Specific dosing schedules are shown in Table 5-1. Tumor volume was measured, mice were weighed, and the data were recorded.

[0180] The experimental indicator is to examine the effect of drugs on tumor growth, and the specific indicator is tumor inhibition rate TGI (%).

[0181] The tumor diameter was measured with a vernier caliper twice a week, and the tumor volume (V) was calculated as follows: V = 1 / 2 × a × b 2 Where a and b represent length and width respectively.

[0182] Calculation of TGI (%): If the tumor did not regress, TGI (%) = [1 - (average tumor volume at the end of dosing for a given treatment group - average tumor volume at the time of grouping for that treatment group) / (average tumor volume at the end of treatment for the vehicle control group - average tumor volume at the time of grouping for the vehicle control group)] × 100%. If the tumor regressed, TGI (%) = [1 - (average tumor volume at the end of dosing for a given treatment group - average tumor volume at the time of grouping for that treatment group) / average tumor volume at the time of grouping for that treatment group] × 100%.

[0183] The experiment ends, the experimental endpoint is reached, or the average tumor volume reaches 2000mm 3 The animals were killed by CO2 anesthesia, and then the tumors were removed by dissection and photographed.

[0184] Experimental data were analyzed and graphed using GraphPad Prism 9.4. Tumor volume data for each group at different time points were statistically analyzed using Dunnett's multiple comparisons test in a two-way ANOVA to assess intergroup differences. Differences in tumor volume between the two groups were analyzed using the t-test, with a p < 0.05 defining the difference as statistically significant.

[0185] Table 5-1. Dosage regimen of drug A combined with drug B in the RL95-2 model

[0186] 3. Experimental Results

[0187] The growth inhibitory effects of Drug A combined with Drug B on the RL95-2 model are shown in Table 5-2 and Figure 8 . The body weight changes of the animals in each group in the RL95-2 model are shown in Figure 9 . In the RL95-2 human endometrial cancer xenograft mouse model, tumor-bearing mice well tolerated treatment with Drug A alone, Drug B alone, and the combination of Drugs A and B. Drug A alone significantly inhibited the growth of xenograft tumors in nude mice, while Drug B alone had no significant inhibitory effect. The combination of Drugs A and B demonstrated a significant tumor-suppressing effect, significantly superior to treatment with either Drug A or Drug B alone.

[0188] Table 5-2. Growth inhibitory effect of drug A combined with drug B on RL95-2 model

[0189] p-value D28: The value obtained by Dunnett analysis using Two-way ANOVA based on the tumor volume of each animal in the different groups, with the solvent group as the control; in addition, statistical analysis was performed using t-test, and the p-values ​​of the drug B_1mg / kg combined with drug A_3mg / kg group were compared with the B_1mg / kg group and the A_3mg / kg group, respectively, were <0.0001 and 0.0270.

[0190] Example 6. In vivo pharmacodynamic study of drug A alone or in combination therapy in a subcutaneous xenograft tumor model of human ovarian cancer cell line OVCAR3 mice

[0191] 1. Test drug

[0192] Drug A: Prepared by the method in Example 1, using physiological saline.

[0193] Drug B: 1'-((7-ethyl-6-carbonyl-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide was prepared by the method disclosed in WO2022223025 and used as Drug B.

[0194] Drug C: Bevacizumab for injection, provided by Suzhou Shengdia Biopharmaceutical Co., Ltd., and prepared with normal saline.

[0195] 2. Experimental instruments and reagents

[0196] 2.1 Instruments

[0197] CO2 incubator (HERAcell-240i, Thermo Fisher Scientific); precision balance (SECURA225D-1CN, Sartorius AG, Germany); standard balance (HZ2002A, Changzhou Tianzhiping Instrument Equipment Co., Ltd.); biological safety cabinet (BSC1300-II-A2, Shandong Xinhua Medical Equipment Co., Ltd.); digital caliper ((0-150) mm / 0.01 mm, Mitutoyo, Japan); pipettes (20-200 μL; 100-1000 μL, Eppendorf) 2.2 Reagents

[0198] RPMI 1640 was purchased from Gibco, catalog number 22400-071; FBS was purchased from Gibco, catalog number 10091148; PBS was purchased from Gibco, catalog number 10010023; trypsin was purchased from Gibco, catalog number 25200056; Matrigel was purchased from Corning, catalog number 354234.

[0199] 3. Experimental operation and data processing

[0200] 3.1 Animals

[0201] NOD SCID mice, 6-8 weeks old, female, were purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd.

[0202] 3.2 Cell culture and cell suspension preparation

[0203] a. Remove one strain of OVCAR3 cells from the cell bank and resuscitate them in RPMI 1640 medium. Place the revived cells in a cell culture flask (label the flask with the cell type, date, and name of the culturer) and culture them in a CO2 incubator (37°C, 5% CO2).

[0204] b. Subculture the cells once a week and continue to culture them in a CO2 incubator. Repeat this process until the cell number meets the in vivo efficacy requirement.

[0205] c. Collect the cultured cells, count them using an automatic cell counter, and resuspend the cells in PBS according to the counting results to prepare a cell suspension (density 10×10 7 / mL), add an equal volume of Matrigel to the cell suspension, mix evenly, and place in an ice box for use.

[0206] 3.3 Cell seeding

[0207] a, Nude mice were marked with disposable ear tags for both mice and rats before inoculation.

[0208] b. Mix the cell suspension thoroughly during inoculation, draw out 0.2-1 mL of cell suspension with a 1 mL syringe, remove any bubbles, and place the syringe on an ice pack until ready to use.

[0209] c. Restrain the NOD SCID mouse with the left hand and disinfect the right side of the nude mouse's back near the right shoulder (inoculation site) with a 75% alcohol cotton ball. Start inoculation 30 seconds later.

[0210] d, The experimental NOD SCID mice were inoculated sequentially (0.1 mL of cell suspension per mouse).

[0211] 3.4 Tumor measurement, grouping, and drug administration in tumor-bearing mice

[0212] a. Tumors were measured and their size was calculated on days 20-30 after inoculation, depending on tumor growth.

[0213] Tumor volume calculation: Tumor volume (mm 3 ) = length (mm) × width (mm) × width (mm) / 2

[0214] b, Tumor-bearing mice were randomly divided into groups according to their weight and tumor size;

[0215] c. According to the grouping results, the test drug was started. The specific dosage and administration schedule are shown in Table 6-1.

[0216] Table 6-1. Dosage and grouping

[0217] a. Dosing volume: 10 μL / g of mouse body weight. Stop dosing if body weight decreases by more than 15% and resume dosing when body weight recovers to within 10%.

[0218] d, Tumors were measured and weighed twice a week after the start of the test drug administration.

[0219] e, Animals were euthanized after the experiment.

[0220] f. Data were processed using GraphPad Prism and other software. The anti-tumor efficacy of the compound was evaluated using TGI (%) and ΔT / ΔC (%). ΔT / ΔC (%) = (T-T0) / (C-C0) × 100, where T and C are the tumor volumes of the treatment group and the vehicle control group at the end of the experiment, respectively, and T0 and C0 are the tumor volumes of the treatment group and the vehicle control group at the beginning of the experiment. Tumor inhibition rate (TGI) (%) = 100-ΔT / ΔC (%). When the tumor regressed, the tumor inhibition rate (TGI) (%) = 100-(T-T0) / T0 × 100

[0221] 4. Experimental results and conclusions

[0222] Tumor-bearing mice tolerated drug A alone or in combination with drugs B or C. The drug A combined with drug B group significantly outperformed the single-drug treatment groups, and the drug A combined with drug C group significantly outperformed the single-drug treatment groups. Furthermore, the drug A combined with drug B and drug C group had the most significant tumor inhibition effect, outperforming the single-drug and combination treatment groups.

[0223] Example 7. In vivo pharmacodynamic study of drug A alone or in combination therapy in a subcutaneous xenograft tumor model of human endometrial cancer cell line RL95-2 mice

[0224] 1. Test drug

[0225] Drug A: Prepared by the method in Example 1, using physiological saline.

[0226] Drug B: 1'-((7-ethyl-6-carbonyl-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide was prepared by the method disclosed in WO2022223025 and used as Drug B.

[0227] Drug C: Bevacizumab for injection, provided by Suzhou Shengdia Biopharmaceutical Co., Ltd., and prepared with normal saline.

[0228] 2. Experimental Purpose

[0229] The in vivo efficacy of compound A as a single agent or in combination therapy was evaluated in a subcutaneous xenograft tumor model of nude mice with human endometrial cancer cell line RL95-2.

[0230] 3. Experimental instruments and reagents

[0231] 3.1 Instruments

[0232] CO2 incubator (HERAcell-240i, Thermo Fisher Scientific)

[0233] Precision balance (SECURA225D-1CN, Sartorius Group, Germany)

[0234] Ordinary balance (HZ2002A, Changzhou Tianzhiping Instrument Equipment Co., Ltd.)

[0235] Biological safety cabinet (BSC1300-II-A2, Shandong Xinhua Medical Instrument Co., Ltd.)

[0236] Digital caliper (0-150 mm / 0.01 mm, Mitutoyo, Japan)

[0237] Pipette (20-200 μL; 100-1000 μL, Eppendorf)

[0238] 3.2 Reagents

[0239] RPMI 1640 was purchased from Gibco, catalog number 22400-071

[0240] FBS was purchased from Gibco, catalog number 10091148;

[0241] PBS was purchased from Gibco, catalog number 10010023;

[0242] Pancreatin was purchased from Gibco, catalog number 25200056;

[0243] Matrigel was purchased from Corning, catalog number 354234.

[0244] 4. Experimental operation and data processing

[0245] 4.1 Animals

[0246] NOD SCID mice, 6-8 weeks old, female, were purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd.

[0247] 4.2 Cell culture and cell suspension preparation

[0248] a. Remove a strain of RL95-2 cells from the cell bank and resuscitate them in RPMI 1640 medium. Place the revived cells in a cell culture flask (label the flask with the cell type, date, and name of the culturer) and culture them in a CO2 incubator (37°C, 5% CO2).

[0249] b. Subculture cells once a week and continue to culture in a CO2 incubator. Repeat this process until the cell number meets the in vivo efficacy requirement.

[0250] c. Collect the cultured cells, count them using an automatic cell counter, and resuspend the cells in PBS according to the counting results to prepare a cell suspension (density 10×10 7 / mL), add an equal volume of Matrigel to the cell suspension, mix evenly, and place in an ice box for use.

[0251] 3.3 Cell seeding

[0252] a, Nude mice were marked with disposable ear tags for both mice and rats before inoculation.

[0253] b. Mix the cell suspension thoroughly during inoculation. Use a 1 mL syringe to draw 0.2-1 mL of cell suspension, remove any bubbles, and place the syringe on an ice pack until ready to use.

[0254] c. Restrain the NOD SCID mouse with the left hand and disinfect the right side of the nude mouse's back near the right shoulder (inoculation site) with a 75% alcohol cotton ball. Start inoculation 30 seconds later.

[0255] d, The experimental NOD SCID mice were inoculated sequentially (0.1 mL of cell suspension per mouse).

[0256] 3.4 Tumor measurement, grouping, and drug administration in tumor-bearing mice

[0257] a. Tumors were measured and their size was calculated on days 20-30 after inoculation, depending on tumor growth.

[0258] Tumor volume calculation: Tumor volume (mm 3 ) = length (mm) × width (mm) × width (mm) / 2

[0259] b, Tumor-bearing mice were randomly divided into groups according to their weight and tumor size;

[0260] c. According to the grouping results, the test drug was started. The specific dosage and administration schedule are shown in Table 7-1.

[0261] Table 7-1. Dosage and grouping

[0262] a. Dosing volume: 10 μL / g of mouse body weight. Stop dosing if body weight decreases by more than 15% and resume dosing when body weight recovers to within 10%.

[0263] d, Tumors were measured and weighed twice a week after the start of the test drug administration.

[0264] e, Animals were euthanized after the experiment.

[0265] f. Data were processed using GraphPad Prism and other software. The anti-tumor efficacy of the compound was evaluated using TGI (%) and ΔT / ΔC (%). ΔT / ΔC (%) = (T-T0) / (C-C0) × 100, where T and C are the tumor volumes of the treatment group and the vehicle control group at the end of the experiment, respectively, and T0 and C0 are the tumor volumes of the treatment group and the vehicle control group at the beginning of the experiment. Tumor inhibition rate (TGI) (%) = 100-ΔT / ΔC (%). When the tumor regressed, the tumor inhibition rate (TGI) (%) = 100-(T-T0) / T0 × 100

[0266] 4. Experimental results and conclusions

[0267] Tumor-bearing mice tolerated drug A alone or in combination with drugs B or C. The drug A combined with drug B group significantly outperformed the single-drug treatment groups, and the drug A combined with drug C group significantly outperformed the single-drug treatment groups. Furthermore, the drug A combined with drug B and drug C group had the most significant tumor inhibition effect, outperforming the single-drug and combination treatment groups.

[0268] Example 8. Clinical Trial of PARP Inhibitor Combined with Anti-B7H4 Antibody Drug Conjugate for the Treatment of Advanced Solid Tumors

[0269] 1. Research Objectives

[0270] Main research objectives:

[0271] To evaluate the safety and tolerability of PARP inhibitor combination therapy in subjects with advanced solid tumors.

[0272] Secondary study objectives:

[0273] 1. Evaluate the PK characteristics of PARP inhibitor combination therapy in subjects with advanced solid tumors;

[0274] 2. Evaluate other safety indicators of PARP inhibitor combination therapy in subjects with advanced solid tumors;

[0275] 3. Evaluate the efficacy of PARP inhibitor combination therapy in subjects with advanced solid tumors;

[0276] 4. Evaluate the PK characteristics of anti-B7H4 antibody-drug conjugate combination therapy in patients with advanced solid tumors;

[0277] 5. Evaluate the immunogenicity of anti-B7H4 antibody-drug conjugates in combination therapy.

[0278] Exploratory research objectives:

[0279] 1. The relationship between exposure and effect;

[0280] 2. Explore biomarkers that predict or influence the efficacy of combination therapy with anti-PARP inhibitors.

[0281] 2. Name of investigational drug:

[0282] (1) Anti-B7H4 Antibody-Drug Conjugates

[0283] Dosage form: Injection (lyophilized powder), Specification: 50 mg / bottle, Manufacturer: Shanghai Hansoh Biopharmaceutical Technology Co., Ltd.

[0284] (2) Poly adenosinediphosphate ribose polymerase (PARP) inhibitors

[0285] 1'-((7-ethyl-6-carbonyl-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide was prepared using the method disclosed in WO2022223025.

[0286] Dosage form: tablets, specifications: 10mg, 40mg, manufacturer: Shanghai Hansoh Biopharmaceutical Technology Co., Ltd.

[0287] 3. Target group:

[0288] ① Patients with recurrent ovarian cancer, fallopian tube cancer, primary peritoneal cancer, or advanced breast cancer confirmed by cytology or histology who have failed or are intolerant to standard treatment;

[0289] ② Patients with advanced solid tumors who have failed or are intolerant to other standard treatments and who are assessed by the investigator to be likely to benefit from the study treatment.

[0290] 4. Dosage regimen:

[0291] The dose-finding phase of this study has the following cohorts:

[0292] 4A: PARP inhibitor combined with anti-B7H4 antibody drug conjugate

[0293] The PARP inhibitor escalation phase begins with a 40 mg QD starting dose. Subjects will receive oral administration once daily starting on C1D1 (Day 1 of Cycle 1), with each 21-day treatment cycle consisting of one treatment cycle. Subjects should fast from 2 hours before to 1 hour after PARP inhibitor administration.

[0294] The B7H4 antibody-drug conjugate will be administered at a starting dose of 3.8 mg / kg Q3W during the escalation phase, with a total of three dose groups (2.8 mg / kg, 3.8 mg / kg, and 4.8 mg / kg Q3W). If the starting dose is not tolerated, the dose will be reduced to 2.8 mg / kg Q3W. Subjects will receive B7H4 antibody-drug conjugate intravenously starting on C1D1 (+3 days), with each 21-day treatment cycle consisting of a 21-day treatment cycle. Starting from C2 (Cycle 2), the interval between each subsequent first intravenous dose and the previous first intravenous dose will be 21 (±3) days.

[0295] 5. Study endpoints:

[0296] Primary study endpoint:

[0297] The maximum tolerated dose (MTD) or maximum applicable dose (MAD) of PARP inhibitor combination therapy.

[0298] Secondary study endpoints:

[0299] 1. PK characteristics of PARP inhibitor combination therapy;

[0300] 2. Safety of PARP inhibitor combination therapy;

[0301] 3. Effectiveness of PARP inhibitor combination therapy (investigator assessment):

[0302] a. Investigator-assessed objective response rate (ORR), disease control rate (DCR), and duration of response (DoR) according to RECIST v1.1 for subjects with target lesions at baseline (all solid tumors except prostate cancer);

[0303] b. Investigator-assessed progression-free survival (PFS) according to RECIST v1.1 (all solid tumors except prostate cancer);

[0304] c. Overall survival (OS);

[0305] d. Investigator-assessed ORR according to RECIST v1.1 and GCIG CA-125 criteria (ovarian cancer only);

[0306] e. Investigator-assessed ORR, DCR, DoR, and radiographic progression-free survival (rPFS) according to RECIST v1.1 (soft tissue) and PCWG3 criteria (bone lesions) (prostate cancer only);

[0307] f. Proportion of subjects with a CA-125 decrease of ≥50% from baseline and time to CA-125 progression (ovarian cancer only);

[0308] g.PSA50 response rate and time to PSA progression (prostate cancer only).

[0309] 6. PK characteristics of combined anti-B7H4 antibody-drug conjugate therapy in patients with advanced solid tumors.

[0310] Exploratory study endpoints:

[0311] 1. Explore the relationship between exposure and effect;

[0312] 2. Explore biomarkers that predict or influence the efficacy of PARP inhibitor combination therapy.

[0313] Example 9. Clinical Trial of PARP Inhibitor Combined with Anti-B7H4 Antibody Drug Conjugate and Bevacizumab for the Treatment of Advanced Solid Tumors

[0314] 1. Research Objectives

[0315] Main research objectives:

[0316] To evaluate the safety and tolerability of PARP inhibitor combination therapy in subjects with advanced solid tumors.

[0317] Secondary study objectives:

[0318] 1. Evaluate the PK characteristics of PARP inhibitor combination therapy in subjects with advanced solid tumors;

[0319] 2. Evaluate other safety indicators of PARP inhibitor combination therapy in subjects with advanced solid tumors;

[0320] 3. Evaluate the efficacy of PARP inhibitor combination therapy in subjects with advanced solid tumors;

[0321] 4. Evaluate the PK characteristics of anti-B7H4 antibody-drug conjugate combination therapy in patients with advanced solid tumors;

[0322] 5. Evaluate the immunogenicity of anti-B7H4 antibody-drug conjugates in combination therapy.

[0323] Exploratory research objectives:

[0324] 1. The relationship between exposure and effect;

[0325] 2. Explore biomarkers that predict or influence the efficacy of combination therapy with anti-PARP inhibitors.

[0326] 2. Name of investigational drug:

[0327] (1) Anti-B7H4 Antibody-Drug Conjugates

[0328] Dosage form: Injection (lyophilized powder), Specification: 50 mg / bottle, Manufacturer: Shanghai Hansoh Biopharmaceutical Technology Co., Ltd.

[0329] (2) Poly adenosinediphosphate ribose polymerase (PARP) inhibitors

[0330] 1'-((7-ethyl-6-carbonyl-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide was prepared using the method disclosed in WO2022223025.

[0331] Dosage form: tablets, specifications: 10mg, 40mg, manufacturer: Shanghai Hansoh Biopharmaceutical Technology Co., Ltd.

[0332] (3) Anti-VEGF antibodies

[0333] Bevacizumab injection, dosage form: sterile solution for injection, specification: 100 mg (4 mL) / bottle, manufacturer: Suzhou Shengdia Biotechnology Co., Ltd.

[0334] (4) Platinum drugs

[0335] Carboplatin injection, properties: Carboplatin for injection is white or off-white freeze-dried loose blocks or powder, manufacturer: Qilu Pharmaceutical Co., Ltd.

[0336] Cisplatin injection, properties: light yellow-green to light yellow to slightly viscous clear liquid, manufacturer: Jiangsu Hausen Pharmaceutical Group Co., Ltd.

[0337] 3. Target group:

[0338] Patients with platinum-sensitive recurrent ovarian cancer:

[0339] ① Histologically confirmed high-grade (poorly differentiated) serous or high-grade (poorly differentiated) endometrioid epithelial ovarian cancer, fallopian tube cancer, or primary peritoneal cancer.

[0340] ②Disease progression or recurrence after 2-3 lines of treatment, and disease progression or recurrence is ≥ 6 months from the completion of the last platinum-containing chemotherapy.

[0341] ③ No systemic anti-tumor treatment has been received since the most recent progression or recurrence.

[0342] ④Previously received at least one PARP inhibitor treatment.

[0343] ⑤ The researcher assesses that the patient is not suitable for tumor reduction surgery (e.g., surgical resection cannot achieve satisfactory tumor reduction), or the subject refuses tumor reduction surgery.

[0344] Newly diagnosed advanced ovarian cancer not receiving systemic anticancer therapy:

[0345] ① Patients with histologically or cytologically confirmed or with predominantly high-grade (poorly differentiated) serous or high-grade (poorly differentiated) endometrioid ovarian cancer, fallopian tube cancer, or primary peritoneal cancer, and FIGO stage III or IV. Patients with histologically classified as carcinosarcoma, undifferentiated, or mucinous cell carcinoma or clear cell carcinoma in epithelial ovarian cancer are not permitted.

[0346] ② The subject has completed or plans to undergo initial tumor cytoreductive surgery, or plans to undergo intermediate tumor cytoreductive surgery.

[0347] ③No previous treatment with PARP inhibitors.

[0348] ④The patients have not received any systemic anti-tumor treatment for advanced ovarian cancer in the past.

[0349] 4. Dosage regimen:

[0350] The dose-finding phase of this study has the following cohorts:

[0351] Cohort 5A: anti-B7H4 antibody-drug conjugate combined with a PARP inhibitor and bevacizumab.

[0352] In this dose-finding process, an anti-B7H4 antibody-drug conjugate was used in combination with bevacizumab and a platinum drug in the induction phase, and an anti-B7H4 antibody-drug conjugate was used in combination with bevacizumab and a PARP inhibitor in the sequential maintenance phase.

[0353] During the induction therapy phase, the day of the subject's first medication is recorded as C1D1 (the first day of the first cycle), and the day of each subsequent medication is the D1 of the next treatment cycle. The medication interval window period is 21±3 days; after completing the induction therapy phase and before entering maintenance therapy, the subject needs to undergo a tumor imaging assessment. If the disease has not progressed, they can enter the maintenance therapy phase. The first dose of PARP inhibitors in the maintenance therapy phase should be within 3 to 6 weeks after the last dose of platinum in the induction therapy phase. The date of the first PARP inhibitor drug treatment in the maintenance therapy phase is defined as D1 of the current visit, and every 21 days thereafter is a treatment cycle. The following is the dosing regimen for various drugs in this cohort:

[0354] The starting dose of the PARP inhibitor is 40 mg QD, with three pre-defined dose groups (20 mg, 40 mg, and 80 mg QD). If the starting dose is not tolerated, the dose is reduced to 20 mg QD. PARP inhibitor treatment should be started 3 to 6 weeks after the completion of platinum-based chemotherapy in the induction phase. Each 21-day treatment cycle is completed, and the drug should be taken orally once daily. Subjects should fast from 2 hours before to 1 hour after PARP inhibitor administration.

[0355] The starting dose of the anti-B7H4 antibody drug conjugate is 3.8 mg / kg every 3 weeks, with a total of three dose groups (2.8 mg / kg, 3.8 mg / kg, and 4.8 mg / kg every 3 weeks). If the starting dose is not tolerated, the dose will be reduced to 2.8 mg / kg every 3 weeks. Subjects will receive intravenous administration of the anti-B7H4 antibody drug conjugate starting on C1D1 (+3 days). Starting from C2, the interval between the first intravenous dose and the previous first intravenous dose will be 21 (±3) days.

[0356] Platinum: Cisplatin 75 mg / m2 at the investigator's option 2Patients with platinum-sensitive recurrent ovarian cancer will receive 6 cycles of platinum-based chemotherapy, or patients with newly diagnosed advanced ovarian cancer will receive 6 to 8 cycles of platinum-based chemotherapy, administered every 3 weeks or at a carboplatin AUC of 5 mg / mL / min every 3 weeks. Patients will receive intravenous platinum-based chemotherapy starting on C1D1 (+3 days). Starting on C2, the interval between the first intravenous dose and the previous first dose will be 21 (±3) days. Investigators may choose cisplatin or carboplatin based on the patient's specific condition. Cisplatin and carboplatin may be used interchangeably during treatment due to tolerability issues, but patients should be fully informed of the risk of cross-sensitivity. If a subject cannot tolerate the drug due to safety reasons, treatment will be discontinued.

[0357] Bevacizumab: All subjects in the dose-escalation phase will receive bevacizumab. Subjects will receive 15 mg / kg intravenously every three weeks. Newly diagnosed ovarian cancer subjects will receive bevacizumab starting on Day 1 (+3 days) and continuing for up to 15 months. Subjects with platinum-sensitive recurrent ovarian cancer will receive bevacizumab starting on Day 1 (+3 days) and continuing until objective disease progression (unless continued after disease progression) or other discontinuation criteria are met.

[0358] In this cohort, intravenous medications are recommended after PARP inhibitor administration. Intravenous medications are recommended to be administered in the following order: bevacizumab first (if applicable), followed by anti-B7H4 antibody-drug conjugates, and finally platinum (if applicable). All drugs are administered continuously until the completion of the pre-specified treatment course, objective disease progression (except for continued treatment after disease progression), or other discontinuation criteria are met.

[0359] 5. Study endpoints:

[0360] Primary study endpoint:

[0361] The maximum tolerated dose (MTD) or maximum applicable dose (MAD) of PARP inhibitor combination therapy.

[0362] Secondary study endpoints:

[0363] 1. PK characteristics of PARP inhibitor combination therapy;

[0364] 2. Safety of PARP inhibitor combination therapy;

[0365] 3. Effectiveness of PARP inhibitor combination therapy (investigator assessment):

[0366] a. Investigator-assessed objective response rate (ORR), disease control rate (DCR), and duration of response (DoR) according to RECIST v1.1 for subjects with target lesions at baseline (all solid tumors except prostate cancer);

[0367] b. Investigator-assessed progression-free survival (PFS) according to RECIST v1.1 (all solid tumors except prostate cancer);

[0368] c. Overall survival (OS);

[0369] d. Investigator-assessed ORR according to RECIST v1.1 and GCIG CA-125 criteria (ovarian cancer only);

[0370] e. Investigator-assessed ORR, DCR, DoR, and radiographic progression-free survival (rPFS) according to RECIST v1.1 (soft tissue) and PCWG3 criteria (bone lesions) (prostate cancer only);

[0371] f. Proportion of subjects with a CA-125 decrease of ≥50% from baseline and time to CA-125 progression (ovarian cancer only);

[0372] g.PSA50 response rate and time to PSA progression (prostate cancer only).

[0373] 6. PK characteristics of combined anti-B7H4 antibody-drug conjugate therapy in patients with advanced solid tumors.

[0374] Exploratory study endpoints:

[0375] 1. Explore the relationship between exposure and effect;

[0376] 2. Explore biomarkers that predict or influence the efficacy of PARP inhibitor combination therapy.

Claims

1. Use of an antibody-drug conjugate and a poly(ADP-ribose) polymerase inhibitor in the preparation of a drug for treating cancer, wherein the structure of the antibody-drug conjugate is shown in formula (I): in: n is 1 to 10, preferably 2 to 8, more preferably 3 to 8, and n is a decimal or an integer; Pc is an anti-B7H4 antibody or an antigen-binding fragment thereof.

2. The use according to claim 1, characterized in that The anti-B7H4 antibody or its antigen-binding fragment comprises: heavy chain HCDR1, HCDR2, HCDR3 as shown in the amino acid sequences of SEQ ID NOs: 01, 02 and 03, respectively, and light chain LCDR1, LCDR2 and LCDR3 as shown in the amino acid sequences of SEQ ID NOs: 04, 05 and 06, respectively.

3. The use according to claim 1 or 2, characterized in that The anti-B7H4 antibody or antigen-binding fragment thereof is selected from a humanized antibody or a fragment thereof.

4. The use according to claim 3, characterized in that The anti-B7H4 antibody or its antigen-binding fragment comprises a heavy chain constant region of human IgG1, IgG2, IgG3 or IgG4 isotype, and a light chain constant region comprising κ or λ; preferably, the anti-B7H4 antibody or its antigen-binding fragment comprises a heavy chain constant region of IgG1 or IgG4 isotype.

5. The use according to claim 3, characterized in that The heavy chain variable region sequence of the anti-B7H4 antibody or antigen-binding fragment thereof is as shown in SEQ ID NO: 07 or a variant thereof, and the light chain variable region sequence is as shown in SEQ ID NO: 08 or a variant thereof.

6. The use according to any one of claims 1 to 5, characterized in that The heavy chain sequence of the anti-B7H4 antibody or antigen-binding fragment thereof is the sequence shown in SEQ ID NO: 09 or a variant thereof, and the light chain sequence is the sequence shown in SEQ ID NO: 10 or a variant thereof.

7. The use according to any one of claims 1 to 6, characterized in that The poly(ADP-ribose) polymerase inhibitor is selected from one or more of Olaparib, Fluzoparib, Niraparib, Pamiparib, Rucaparib, Talazoparib, Veliparib, Senaparib, CEP-8983, BGB-290 or 1'-((7-ethyl-6-carbonyl-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide.

8. The use according to any one of claims 1 to 7, characterized in that The combination has additive or synergistic pharmacodynamic effects.

9. The use according to any one of claims 1 to 8, characterized in that The antibody-drug conjugate and the poly(ADP-ribose) polymerase inhibitor are contained in different preparations as active ingredients, respectively, and are administered simultaneously or at different times.

10. The use according to any one of claims 1 to 8, characterized in that The antibody drug conjugate and the poly(ADP-ribose) polymerase inhibitor are contained in a single preparation as active ingredients and administered.

11. The use according to any one of claims 1 to 10, further combined with an anti-VEGF antibody.

12. The use according to claim 11, wherein the anti-VEGF antibody is selected from Bevacizumab, Ranibizumab, Sevacizumab, Suvemcitug, Varisacumab, CMAB-801, LYN-00101.

13. The use according to claim 11 or 12, wherein the antibody-drug conjugate, poly(ADP-ribose) polymerase inhibitor, and anti-VEGF antibody are contained in different preparations as active ingredients, respectively, and are administered simultaneously, concurrently, sequentially, continuously, alternately or separately.

14. The use according to any one of claims 11 to 13, wherein the anti-VEGF antibody is administered at a dosage of 1.0 mg / kg to 100 mg / kg, preferably 1.0 mg / kg to 40 mg / kg, more preferably 1.0 mg / kg to 30 mg / kg, and the administration frequency is once a week, once every two weeks, once every three weeks or once every four weeks.

15. The use according to any one of claims 1 to 14, characterized in that The dosage of the antibody drug conjugate is 0.1 mg / kg to 20.0 mg / kg, preferably 1.0 mg / kg to 15.0 mg / kg; the administration frequency is once a week, once every two weeks, once every three weeks or once every four weeks.

16. The use according to any one of claims 1 to 15, characterized in that The dosage of the poly(ADP-ribose) polymerase inhibitor is 1-500 mg / kg, preferably 10 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 35 mg / kg, 40 mg / kg, 45 mg / kg, 50 mg / kg, 55 mg / kg, 60 mg / kg, 65 mg / kg, 70 mg / kg, 75 mg / kg, 80 mg / kg, 85 mg / kg, 90 mg / kg, 95 mg / kg, 100 mg / kg, 110 mg / kg, 120 mg / kg, 130 mg / kg, 140 mg / kg, 150 mg / kg, 160 mg / kg, 170 mg / kg, 180 or 500 mg / kg; the frequency of administration is once a day, twice a day, three times a day, once every two days or once every three days.

17. The use according to any one of claims 1 to 16, characterized in that The cancer is a solid tumor. Preferably, the cancer is selected from uterine cancer, breast cancer, ovarian cancer, pancreatic cancer, bile duct cancer, fallopian tube cancer, gastric cancer, colorectal cancer, primary peritoneal cancer, prostate cancer, and lung cancer; more preferably, uterine cancer, breast cancer, and ovarian cancer.

18. The use according to any one of claims 1 to 17, characterized in that The cancer is selected from cervical cancer, endometrial cancer, triple-negative breast cancer, ovarian epithelial cancer, epithelial ovarian cancer, platinum-sensitive recurrent ovarian cancer, intrahepatic bile duct carcinoma, and extrahepatic bile duct carcinoma; preferably, endometrial cancer, triple-negative breast cancer, and ovarian epithelial cancer; further preferably, the ovarian epithelial cancer is selected from high-grade serous ovarian cancer, low-grade serous ovarian cancer, intrauterine-like ovarian cancer, mucinous ovarian cancer, and ovarian clear cell carcinoma.

19. A pharmaceutical composition comprising the antibody-drug conjugate according to any one of claims 1 to 18 and a poly(ADP-ribose) polymerase inhibitor, characterized in that: Contains one or more pharmaceutically acceptable excipients, diluents or carriers.

20. A pharmaceutical combination comprising an antibody drug conjugate as defined in any one of claims 1 to 18 and a pharmaceutically acceptable excipient thereof, and a poly(ADP-ribose) polymerase inhibitor as defined in any one of claims 1 to 18 and a pharmaceutically acceptable excipient thereof.

21. A pharmaceutical composition comprising the antibody-drug conjugate according to any one of claims 11 to 18, a poly(ADP-ribose) polymerase inhibitor and an anti-VEGF antibody, characterized in that: Contains one or more pharmaceutically acceptable excipients, diluents or carriers.

22. A pharmaceutical combination comprising an antibody-drug conjugate as defined in any one of claims 1 to 18 and a pharmaceutically acceptable excipient thereof, a poly(ADP-ribose) polymerase inhibitor as defined in any one of claims 1 to 18 and a pharmaceutically acceptable excipient thereof, and an anti-VEGF antibody as defined in any one of claims 11 to 18 and a pharmaceutically acceptable excipient thereof.

23. A method for treating and / or preventing cancer, comprising: An effective amount of an antibody drug conjugate as defined in any one of claims 1 to 18, a poly(ADP-ribose) polymerase inhibitor, and optionally an anti-VEGF antibody as defined in any one of claims 11 to 18 is administered to a patient in combination, the combined administration being simultaneous, concurrent, sequential, continuous, alternating or separate administration.

24. A method for treating and / or preventing cancer, the method comprising administering to a patient an antibody-drug conjugate as defined in any one of claims 1 to 18, an anti-VEGF antibody as defined in any one of claims 11 to 18, and a platinum drug in combination during an induction therapy phase, wherein the combined administration may be simultaneous, concurrent, sequential, continuous, alternating, or separate administration, and in a maintenance therapy phase after the induction therapy, administering to the patient an effective amount of an antibody-drug conjugate as defined in any one of claims 1 to 18, a poly(ADP-ribose) polymerase inhibitor, and an anti-VEGF antibody as defined in any one of claims 11 to 18 in combination, wherein the combined administration may be simultaneous, concurrent, sequential, continuous, alternating, or separate administration.

25. The method according to claim 24, wherein the platinum drug is selected from carboplatin, cisplatin, oxaliplatin, nedaplatin, lobaplatin, satraplatin, cycloplatin, miboplatin, enloplatin, iproplatin, dicycloplatin; preferably carboplatin and / or cisplatin.

26. The method according to claims 23 to 25, characterized in that The cancer is a solid tumor. Preferably, the cancer is selected from uterine cancer, breast cancer, ovarian cancer, pancreatic cancer, bile duct cancer, fallopian tube cancer, gastric cancer, colorectal cancer, primary peritoneal cancer, prostate cancer, and lung cancer; more preferably, uterine cancer, breast cancer, and ovarian cancer.

27. The method according to claims 23 to 26, characterized in that The cancer is selected from cervical cancer, endometrial cancer, triple-negative breast cancer, ovarian epithelial cancer, platinum-sensitive recurrent ovarian cancer, intrahepatic bile duct carcinoma, and extrahepatic bile duct carcinoma; preferably, endometrial cancer, triple-negative breast cancer, and ovarian epithelial cancer; further preferably, the ovarian epithelial cancer is selected from high-grade serous ovarian cancer, low-grade serous ovarian cancer, high-grade endometrioid epithelial ovarian cancer, endometrioid ovarian cancer, mucinous ovarian cancer, and ovarian clear cell carcinoma.

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