Anti-b7h3 antibody-eribulin conjugate and use thereof

Abstract

An anti-B7H3 antibody-eribulin conjugate of the following formula, a pharmaceutically acceptable salt or solvate thereof, a pharmaceutical composition comprising same, and use thereof. The conjugate is used for preparing a medicament for treating and / or preventing B7H3-mediated diseases or disorders, particularly diseases or disorders with positive B7H3 expression, such as cancer. The anti-B7H3 antibody-eribulin conjugate demonstrates high antigen affinity, strong cellular internalization, significantly improved drug release efficiency, and enhanced killing and inhibition abilities against tumor cells, while maintaining linker stability in plasma and low cytotoxicity. In the formula, TL represents the anti-B7H3 antibody, and n is an integer or decimal of 1-20.

Description

An anti-B7H3 antibody-eribulin conjugate and its application

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to patent application number “CN202410406439.8” filed on April 3, 2024, with the invention name “An anti-B7H3 antibody-eribulin conjugate and its application”, and patent application number “CN202410966155.4” filed on July 18, 2024, with the invention name “An anti-B7H3 antibody-eribulin conjugate and its application”, the contents of which are incorporated herein by reference in their entirety. Technical Field

[0003] The present application relates to the technical field of antibody-drug conjugates, and in particular, to an anti-B7H3 antibody-eribulin conjugate and its application. Background Art

[0004] Antibody-drug conjugates (ADCs) are produced by coupling a bioactive small molecule drug to a monoclonal antibody (mAb) via a linker. Currently, the vast majority of ADCs are composed of antibodies targeting tumor antigens coupled to highly cytotoxic small molecule drugs via linkers. Leveraging the specific binding properties of antibodies to target antigens, small molecule drugs are delivered to tumor cells, thereby exerting their tumor-killing effects. There are also cases where proteins and peptides with similar specific binding properties as antibodies, such as ligands, antigens, and peptides, are coupled to toxins via linkers.

[0005] B7H3, also known as CD276, is a member of the B7 family and a type I transmembrane protein. It consists of an amino-terminal signal peptide, an extracellular immunoglobulin-like variable (IgV) and constant (IgC) region, a transmembrane region, and a 45-amino acid cytoplasmic tail. Currently, B7H3 exists in two main splice forms: B7H3a and B7H3b. The extracellular segment of B7H3a is composed of two immunoglobulin domains, IgV-IgC, and is also known as 2IgB7H3. The extracellular segment of B7H3b is composed of four immunoglobulin domains, IgV-IgC-IgV-IgC, and is also known as 4IgB7H3.

[0006] B7H3 is considered a co-stimulatory molecule for T cell activation and IFN-γ production in immune responses. It is a key immune checkpoint protein that is not expressed or expressed at very low levels in normal tissues and cells, but is highly expressed in a variety of tumor tissues. It can promote tumor proliferation, metastasis, and recurrence, and is closely associated with tumor progression, patient survival, and prognosis. Clinically, B7H3 has been reported to be overexpressed in many cancer types, particularly melanoma, leukemia, breast cancer, non-small cell lung cancer, renal cancer, sarcoma, urothelial carcinoma, colorectal cancer, prostate cancer, glioblastoma multiforme, ovarian cancer, and pancreatic cancer. Therefore, B7H3 is considered a new tumor marker and potential therapeutic target.

[0007] Eribulin is a potent mitotic inhibitor. As a toxin in an antibody-drug conjugate, it possesses a large molecular weight, complex structure, and good hydrophilicity. The large molecular weight requires selection of a linker of appropriate length. Complex structures can result in varying efficiency of enzymatic cleavage due to steric hindrance. High hydrophilicity is therefore more suitable for preparing high-conjugation-ratio conjugates of a single antibody molecule coupled to multiple toxins.

[0008] Linkers are one of the key factors affecting drug stability and therapeutic window. First, the linker needs to have a certain degree of stability so that it can ensure the integrity of the ADC during blood circulation before reaching the tumor cells, avoid premature release of toxins leading to off-target toxicity, and affect the therapeutic window of the ADC drug. After entering the target cells, the linker must ensure the effective release of the toxin to exert its killing effect. PEG, a commonly used component of a linker, has good hydrophilicity and stability. The commonly used PEG length is 2 to 8, but its light absorption is not obvious and its synthesis is more complicated. Peptide linkers are a more common cleavable linker. As of September 2022, more than ten ADC drugs have been approved for marketing. Most of them are achieved by coupling the toxin to the cysteine ​​residue of the monoclonal antibody through a peptide linker. The peptide can be cleaved by the tissue protease of the tumor cell to release the toxin, producing an anti-tumor effect.

[0009] There is still a need in the art for antibody-drug conjugates that can specifically bind to antigens, effectively kill tumor cells, and have low cytotoxicity. Summary of the Invention

[0010] The present application provides an anti-B7H3 antibody-eribulin conjugate and its application, which has high antigen affinity, strong cellular internalization, significantly improved drug release efficiency, stronger tumor cell killing and inhibition capabilities, and the linker is stable in plasma and has low cytotoxicity; it has good application prospects in cancer treatment.

[0011] First, the present application provides an anti-B7H3 antibody-eribulin conjugate, or a pharmaceutically acceptable salt or solvate thereof. The structure of the anti-B7H3 antibody-eribulin conjugate is shown below:

[0012] Wherein, TL is an anti-B7H3 antibody, and n is an integer or decimal from 1 to 20.

[0013] Said n can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, and decimals between each integer. Said n is preferably an integer or decimal of 2 to 8, more preferably an integer or decimal of 3 to 6, 3 to 5, or 4 to 5, such as n=3, 3.5, 4, 4.25, 4.5, or 5.

[0014] The anti-B7H3 antibody has a heavy chain variable region HCVR as shown in SEQ ID NO: 1, a light chain variable region LCVR as shown in SEQ ID NO: 2, a heavy chain constant region as shown in SEQ ID NO: 3, and a light chain constant region as shown in SEQ ID NO: 4. Preferably, the anti-B7H3 antibody is a monoclonal antibody.

[0015] The present application also provides a pharmaceutical composition comprising the anti-B7H3 antibody-eribulin conjugate as described above, a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable excipient, diluent or carrier.

[0016] Optionally, the pharmaceutical composition of the present application further comprises one or more other active ingredients that are required for the specific indication being treated, preferably those having complementary activities that do not adversely affect each other.

[0017] The present application also provides the use of the anti-B7H3 antibody-eribulin conjugate, a pharmaceutically acceptable salt or solvate thereof, or the pharmaceutical composition described above in the preparation of a medicament for treating and / or preventing a B7H3-mediated disease or condition. Preferably, the disease or condition is a disease or condition in which B7H3 is positively expressed. Preferably, the disease or condition is cancer.

[0018] The present application provides a method for treating and / or preventing a B7H3-mediated disease or condition, comprising administering to a subject in need thereof an anti-B7H3 antibody-eribulin conjugate, a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition as described above. Preferably, the disease or condition is a disease or condition in which B7H3 is positively expressed. Preferably, the disease or condition is cancer.

[0019] In some embodiments, the cancer described herein may be a metastatic or recurrent cancer.

[0020] In some embodiments, the cancer described herein is a solid tumor or a hematological tumor selected from breast cancer (such as triple-negative breast cancer, HER-2 positive breast cancer, HER-2 positive triple-negative breast cancer, Luminal A breast cancer, Luminal B breast cancer), ovarian cancer (such as ovarian epithelial tumor, ovarian sex cord-stromal tumor, ovarian germ cell tumor, ovarian metastatic tumor), ovarian teratoma, cervical cancer, uterine cancer, prostate cancer (such as metastatic castration-resistant prostate cancer), kidney cancer, urethral cancer, testicular cancer, bladder cancer, liver cancer, gastric cancer, head and neck cancer, endometrial cancer, salivary gland cancer, esophageal cancer, esophageal cancer, lung cancer (such as non-small cell lung cancer, small cell lung cancer, degenerative lung cancer, squamous cell lung cancer), colon cancer, rectal cancer, colorectal cancer, bone cancer, skin cancer, thyroid cancer, pancreatic cancer, melanoma, neural tumors (such as neuroblastoma), glioma Solid tumors or hematologic tumors include gliomas (such as gliomas, multiforme gliomas, glioblastomas), sarcomas (such as rhabdomyosarcoma, fibrosarcoma, embryonal rhabdomyosarcoma, alveolar rhabdomyosarcoma, multiforme rhabdomyosarcoma, sclerosing rhabdomyosarcoma), squamous cell carcinoma (such as head and neck squamous cell carcinoma, lung squamous cell carcinoma), lymphomas (such as non-Hodgkin's lymphoma, diffuse large B-cell lymphoma), myeloma (such as multiple myeloma) and leukemias (such as chronic leukemia, acute leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, acute lymphocytic leukemia, chronic myeloid leukemia, chronic myeloid leukemia).

[0021] In some embodiments, preferably, the cancer described in the present application is selected from lung cancer (such as non-small cell lung cancer, degenerative lung cancer, squamous cell lung carcinoma), sarcoma (such as rhabdomyosarcoma, fibrosarcoma), ovarian cancer, ovarian teratoma, breast cancer (such as triple-negative breast cancer, HER-2 positive breast cancer, HER-2 positive triple-negative breast cancer).

[0022] Optionally, the method for treating and / or preventing a B7H3-mediated disease or condition described herein further comprises administering to the subject one or more other active ingredients, or administering one or more other therapies. The active ingredients are those required for the specific indication being treated, preferably those having complementary activities that do not adversely affect each other, and the one or more other active ingredients can be co-administered with the anti-B7H3 antibody-eribulin conjugate, its pharmaceutically acceptable salt or solvate, or the pharmaceutical composition as described above, or administered sequentially. The one or more other therapies refer to surgical treatment, radiotherapy, electric field therapy, etc.; they can be co-administered with the administration of the anti-B7H3 antibody-eribulin conjugate, its pharmaceutically acceptable salt or solvate, or the pharmaceutical composition as described above, and / or the administration of one or more other active ingredients, or administered separately.

[0023] The anti-B7H3 antibody-eribulin conjugates, and pharmaceutically acceptable salts or solvates thereof, described herein, have strong killing and inhibitory abilities against a variety of tumor cells. Therefore, the anti-B7H3 antibody-eribulin conjugates, and pharmaceutically acceptable salts or solvates thereof, described herein, are suitable for use in patients with tumors that are sensitive to eribulin. The anti-B7H3 antibody-eribulin conjugates, and pharmaceutically acceptable salts or solvates thereof, described herein, also exhibit significant anti-tumor effects in a DS-7300a-resistant xenograft model. Therefore, they are suitable for use in patients with tumors that are resistant to camptothecins and recombinant allosteric human tumor necrosis factor-related apoptosis-inducing ligands, such as those resistant to antibody-drug conjugates with DXD or CPT as payloads. The anti-B7H3 antibody-eribulin conjugate described in the present application, its pharmaceutically acceptable salt or solvate can be used for tumor patients resistant to topoisomerase 1 inhibitors, such as late-stage patients who are resistant to topoisomerase 1 inhibitors after receiving B7H3-ADC drug treatment based on topoisomerase 1 inhibitors.

[0024] The anti-B7H3 antibody-eribulin conjugate described in the present application, and its pharmaceutically acceptable salt or solvate are stable in human / monkey plasma. Experiments have shown that it has low toxicity and good tolerance in cynomolgus monkeys, and has a wider therapeutic window. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] In order to more clearly illustrate the specific implementation methods of the present application or the technical solutions in the prior art, the following is a brief introduction to the drawings required for use in the specific implementation methods or the description of the prior art. Obviously, the drawings described below are some implementation methods of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without any creative work.

[0026] Figures 1 and 2 show the in vitro anti-tumor experimental results of the anti-B7H3 antibody-eribulin conjugate ILB-3101 described in this application.

[0027] Figures 3 to 6 show the in vivo anti-tumor experimental results of the anti-B7H3 antibody-eribulin conjugate ILB-3101 described in this application.

[0028] FIG7 shows the pharmacokinetic experimental results of the anti-B7H3 antibody-eribulin conjugate ILB-3101 described in the present application in cynomolgus monkeys. DETAILED DESCRIPTION

[0029] References to embodiments of the present application will now be provided in detail, one or more examples of which are described below. Each example is provided to illustrate, not to limit, the present application. Indeed, it will be apparent to those skilled in the art that various modifications and variations may be made to the present application without departing from the scope or spirit of the present application. For example, features illustrated or described as part of one embodiment may be used in another embodiment to produce further embodiments.

[0030] Unless otherwise indicated, all terms (including technical and scientific terms) used to disclose this application have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. By way of further guidance, the following definitions are provided to better understand the teachings of this application. The interpretation of the terms used in the specification of this application is only for the purpose of describing specific embodiments and is not intended to limit this application.

[0031] All documents mentioned in this application are cited as references in this application, just as each document is cited as a reference individually. Unless they conflict with the invention purpose and / or technical solution of this application, the cited documents involved in this application are cited in their entirety and for all purposes. When cited documents are involved in this application, the definitions of relevant technical features, terms, nouns, phrases, etc. in the cited documents are also cited. When cited documents are involved in this application, the examples and preferred embodiments of the cited relevant technical features may also be incorporated into this application as references, but are limited to the ability to implement this application. It should be understood that when the cited content conflicts with the description in this application, the present application shall prevail or be adaptively amended according to the description in this application.

[0032] The terms "and / or", "or / and", and "and / or" used herein include any one of two or more related listed items, and also include any and all combinations of the related listed items, wherein the arbitrary and all combinations include any combination of two related listed items, any more related listed items, or all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", "or / and", and "and / or", it should be understood that in this application, the technical solution undoubtedly includes technical solutions connected by "logical and" and also undoubtedly includes technical solutions connected by "logical or". For example, "A and / or B" includes three parallel solutions of A, B and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, the technical solution of all being connected by "logical OR"), and also includes any and all combinations of A, B, C, and D, that is, the combination of any two or any three of A, B, C, and D, and also includes the four-item combination of A, B, C, and D (that is, the technical solution of all being connected by "logical AND").

[0033] As used in this application, the terms "comprising," "including," and "comprising" are synonymous and are inclusive or open-ended and do not exclude additional, unrecited members, elements, or method steps.

[0034]

[0014] The recitation of numerical ranges by endpoints herein includes all numbers and fractions subsumed within that range, as well as the recited endpoints.

[0035] Concentration values ​​used in this application include fluctuations within a certain range. For example, fluctuations within a certain precision range are permitted. For example, for a 2% value, fluctuations within ±0.1% are permitted. For larger values ​​or values ​​that do not require overly precise control, greater fluctuations are permitted. For example, for 100mM, fluctuations within ±1%, ±2%, ±5%, and so on are permitted. Regarding molecular weight, fluctuations within ±10% are permitted.

[0036] In this application, descriptions such as "multiple" and "multiple" refer to a quantity greater than or equal to 2 unless otherwise specified.

[0037] In this application, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0038] In this application, "preferred", "better", "more preferred", and "suitable" are only used to describe implementation methods or examples with better effects. It should be understood that they do not constitute a limitation on the scope of protection of this application. In this application, "optionally", "optional", and "optional" mean that they are optional, that is, they refer to either of the two parallel schemes of "yes" or "no". If multiple "optional" items appear in a technical solution, unless otherwise specified and there is no contradiction or mutual restriction, each "optional" item is independent.

[0039] As used herein, the term "eribulin" refers to a synthetic analog of halichondria B, a macrocyclic compound originally isolated from the sea sponge Halichondria okadais, having the CAS number 253128-41-5. Eribulin is a microtubule dynamics inhibitor that is believed to bind to tubulin and cause cell cycle arrest in the G2 / M phase by inhibiting mitotic spindle assembly. The term "eribulin mesylate," CAS number 441045-17-6, refers to the mesylate salt of eribulin, which is marketed under the trade name Halaven TM sell.

[0040] The term "pharmaceutically acceptable salt" refers to a salt of the anti-B7H3 antibody-eribulin conjugate described herein, meaning that an acceptable salt is administered to a patient (e.g., a mammal) (for a given dosage regimen, it is a salt containing a counterion with acceptable mammalian safety). Such salts can be derived from pharmaceutically acceptable inorganic or organic bases, as well as pharmaceutically acceptable inorganic or organic acids. The conjugate of the present application contains at least one amino group and can therefore form salts with acids. Non-limiting examples of pharmaceutically acceptable salts include: hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, citrate, acetate, succinate, ascorbate, oxalate, nitrate, sorbate, hydrogen phosphate, dihydrogen phosphate, salicylate, hydrogen citrate, tartrate, maleate, fumarate, formate, benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, and p-toluenesulfonate. A particularly preferred pharmaceutically acceptable salt is eribulin mesylate.

[0041] The term "solvate" refers to a combination of the conjugate of the present application and a solvent molecule formed by solvation. In some cases, the solvate refers to a hydrate, i.e., the solvent molecule is a water molecule, and the combination of the conjugate of the present application and water forms a hydrate.

[0042] The term "anti-B7H3 antibody" refers to a protein that exhibits binding specificity for the B7H3 antigen. Classical antibody molecules are usually tetramers composed of two identical heavy chains and two identical light chains interconnected by disulfide bonds. Based on the conservative differences in the amino acid sequence, the heavy chain and light chain are divided into a variable region (V) located at the amino terminus and a constant region (C) located at the carboxyl terminus. The variable region is used to recognize and bind to the antigen, and the constant region (such as the Fc fragment) is used to initiate downstream effects. Within the variable region of the heavy chain and light chain, there are three local areas with a higher degree of variation in amino acid composition and arrangement sequence, which are key locations for antibody-antigen binding and are therefore also called complementarity determining regions (CDRs). A monoclonal antibody (mAb) refers to a highly uniform antibody produced by a single B cell clone (whose gene can only encode one antibody) and targets only a specific antigen epitope.

[0043] The term "pharmaceutically acceptable excipient, diluent or carrier" includes any material that, when combined with the active ingredient, allows the ingredient to retain biological activity and is non-reactive with the subject's immune system.

[0044] The term "cancer" refers to a physiological condition or disease characterized by unregulated cell growth, including solid tumors or blood tumors, metastatic, recurrent cancers, which can be selected from solid tumors or blood tumors, breast cancer (such as triple-negative breast cancer, HER-2 positive breast cancer, HER-2 positive triple-negative breast cancer, luminal A breast cancer, luminal B breast cancer), ovarian cancer (such as ovarian epithelial tumors, ovarian sex cord-stromal tumors, ovarian germ cell tumors, ovarian metastatic tumors), ovarian teratoma, cervical cancer, uterine cancer, prostate cancer (such as metastatic castration-resistant prostate cancer), kidney cancer, urethral cancer, testicular cancer, bladder cancer, liver cancer, gastric cancer, head and neck cancer, endometrial cancer, salivary gland cancer, esophageal cancer, esophageal cancer, lung cancer (such as non-small cell lung cancer, small cell lung cancer, degenerative lung cancer, squamous cell lung cancer), colon cancer, rectal cancer, colorectal cancer, bone cancer, skin cancer, thyroid cancer, pancreatic cancer, melanoma, neural tumors (such as neuroblastoma), glioma Solid tumors or hematologic tumors include gliomas (such as gliomas, multiforme gliomas, glioblastomas), sarcomas (such as rhabdomyosarcoma, fibrosarcoma, embryonal rhabdomyosarcoma, alveolar rhabdomyosarcoma, multiforme rhabdomyosarcoma, sclerosing rhabdomyosarcoma), squamous cell carcinoma (such as head and neck squamous cell carcinoma, lung squamous cell carcinoma), lymphomas (such as non-Hodgkin's lymphoma, diffuse large B-cell lymphoma), myeloma (such as multiple myeloma) and leukemias (such as chronic leukemia, acute leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, acute lymphocytic leukemia, chronic myeloid leukemia, chronic myeloid leukemia).

[0045] The term "one or more other active ingredients" includes, but is not limited to, antiviral agents, antibiotics, analgesics, corticosteroids, steroids, oxygen, antioxidants, COX inhibitors, cardioprotectants, metal chelators, IFN-γ and / or NSAIDs, viral cancer vaccines (e.g., adenoviral vectors encoding cancer-specific antigens), bacterial cancer vaccines (e.g., non-pyrogenic E. coli expressing one or more cancer-specific antigens), yeast cancer vaccines, N-803 (also known as ALT-803, ALTOR Biosciences), chemotherapeutic drugs, antibodies (e.g., that bind to tumor-associated antigens or patient-specific tumor neoantigens), stem cell transplants (e.g., allogeneic or autologous), and tumor-targeted cytokines (e.g., NHS-IL12, IL-12 conjugated to a tumor-targeted antibody or fragment thereof).

[0046] The term "one or more other therapies" includes, but is not limited to, surgical therapy, radiation therapy, electric field therapy, and the like.

[0047] The terms "subject" and "patient" are used interchangeably herein to refer to any animal, such as any mammal, including but not limited to humans, non-human primates, rodents, dogs, cats, chimpanzees, orangutans, monkeys, gibbons, macaques, marmosets, cynomolgus monkeys, pigs, horses, pandas, and elephants, among others.

[0048] The term "treat" refers to any improvement in the consequences of any disease, such as prolonged survival, less morbidity, and / or reduced side effects of alternative treatment modalities. As is readily understood in the art, complete eradication of the disease is preferred, but not a necessary condition for therapeutic action. As used herein, "treat" refers to administering the conjugate, pharmaceutically acceptable salt, or solvate to a subject (e.g., a patient). Treatment can be curing, healing, alleviating, alleviating, altering, curing, improving, attenuating, improving, or affecting a condition, a symptom of a condition, or a predisposition to a disease, such as cancer.

[0049] The term "prevention" generally refers to an approach to obtaining a beneficial or desired effect, including, but not limited to, a prophylactic benefit. For prophylactic benefit, a pharmaceutical composition can be administered to a patient at risk for developing a particular disease, or to a patient reporting one or more physiological symptoms of a disease, even if the disease has not yet been diagnosed.

[0050] The pharmaceutical compositions of the present application can be administered by any route, as will be understood by those skilled in the art. In some embodiments, the pharmaceutical compositions of the present application are administered intravenously.

[0051] The embodiments of the present application will be described in detail below with reference to the examples. It should be understood that these examples are intended to illustrate the present application only and are not intended to limit the scope of the present application. The experimental methods for which specific conditions are not specified in the following examples are preferably referred to the guidance provided in the present application, and can also be based on the experimental manuals or conventional conditions of this area, or other experimental methods known in the art, or according to the conditions recommended by the manufacturer.

[0052] In the following specific examples, the measured parameters of raw material components may have slight deviations within the range of weighing accuracy unless otherwise specified. For temperature and time parameters, acceptable deviations caused by instrument testing accuracy or operational accuracy are allowed.

[0053] Example 1 Preparation of anti-B7H3 antibodies

[0054] Based on the anti-B7H3 antibody sequence in Chinese patent application CN113402610A, which was published on September 17, 2021, the expression and purification of monoclonal antibodies were further carried out.

[0055] Unless otherwise specified, all antibodies in this application can be expressed using the commercial vector pTT5, and the antibody sequence can be inserted between the EcoR I and Hind III restriction enzyme sites after its promoter. The nucleic acid sequence including the signal peptide before the antibody light chain or heavy chain sequence can be selected from the following sequences:

[0056] The EcoR I restriction enzyme cleavage site and the start amino acid codon are underlined in the above sequence.

[0057] The antibody light chain or heavy chain sequence is followed by:

[0058] TGAAAGCTT (which is the stop codon and Hind III restriction site).

[0059] The antibody amino acid sequence is optimized into a nucleotide sequence and then inserted between the above-mentioned signal peptide and terminator.

[0060] Heavy chain H00 sequence:

[0061] Light chain L00 sequence:

[0062] The anti-B7H3 antibody is clone 7B7, and the antibody with light chain L00 and heavy chain H00 is 7B7.

[0063] The above-mentioned antibody light and heavy chains were constructed into vectors, and the plasmids were extracted. Suspension-acclimated CHO-K1 cells were revived in OPM-CD TransCHO medium (manufacturer: OPM, catalog number P83059) and cultured to a density of 2 million cells / mL with a viability of over 95% in a volume of 1000 mL. 0.5 mg of the light chain plasmid and 0.5 mg of the heavy chain plasmid were combined and dissolved in 10 mL of culture medium. 3 mg of PEI (manufacturer: Polyscience, catalog number: 24765-1) dissolved in culture medium was diluted in 10 mL of culture medium. The plasmid and PEI solutions were mixed and incubated at room temperature for 10 minutes. The solution was then added dropwise to 1000 mL of cell culture medium. After incubation at 37°C for 5 days, the supernatant was harvested by centrifugation at 12,000 g for 15 minutes. The supernatant was purified using HiTrap Mabselect SuRe and eluted with 50 mM acetic acid. The neutralized peak was then filtered using a 30 kD ultrafiltration tube (Merck, Cat. No. UFC9030) and replaced with PBS pH 7.4. The antibody was measured for absorbance at 280 nm, and the concentration was calculated by dividing the absorbance by the theoretical extinction coefficient.

[0064] Example 2 Synthesis of Linker-Eribulin

[0065] Based on Chinese patent application number CN202310853999.3 filed on July 12, 2023, and international patent application PCT / CN2023 / 121374 filed on September 26, 2023, the synthesis of linker-Eribulin Mc-VAGGFG-PAB-Eribulin was carried out.

[0066] The structure of Mc-VAGGFG-PAB-Eribulin (LK-322022) is as follows:

[0067] The synthetic route of compound LK-322022 is:

[0068] Synthesis of Compound 2: Commercially available compound 1 (100 mg, 0.244 mmol) was dissolved in 3 mL of tetrahydrofuran, cooled to 0°C, and HOSU (33.7 mg, 0.293 mmol) and DCC (60.3 mg, 0.293 mmol) were added. The reaction was allowed to react at room temperature for 2 hours. After completion of the reaction, the reaction mixture was filtered and the filtrate was concentrated to obtain the crude product, which was directly used in the next reaction. ESI-MS m / z: 508 (M+H).

[0069] Synthesis of Compound 4: Compound 2 (crude, 0.244 mmol) and commercially available compound 3 (82 mg, 0.244 mmol) were mixed in 3 mL of N,N-dimethylformamide. N,N-diisopropylethylamine (126 mg, 0.976 mmol) was added and allowed to react at room temperature for 2 hours. After completion of the reaction, the solvent was removed by distillation under reduced pressure. The crude product was slurried with 10 mL of water and filtered to obtain the crude title compound 4 (200 mg, yield: 100%). ESI-MS m / z: 729 (M+H).

[0070] Synthesis of Compound 6: Compound 4 (200 mg, 0.274 mmol) was dissolved in 5 mL of N,N-dimethylformamide, cooled to 0°C, and compound 5 (41 mg, 0.329 mmol), N,N-diisopropylethylamine (107 mg, 0.823 mmol), and DEPBT (124 mg, 0.412 mmol) were added sequentially. The mixture was reacted at room temperature for 4 hours. After completion of the reaction, the reaction mixture was monitored by liquid chromatography-mass spectrometry. The reaction solution was quenched with 10 mL of aqueous ammonium chloride and extracted three times with ethyl acetate (10 mL each). The organic phases were separated, combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography [dichloromethane / methanol = 88:12 (v / v)] to give the title compound 6 (100 mg, yield: 43.8%). ESI-MS m / z: 834 (M+H).

[0071] Synthesis of Compound 7: Compound 6 (100 mg, 0.120 mmol) was dissolved in 3 mL of N,N-dimethylformamide, and piperidine (31 mg, 0.360 mmol) was added. The mixture was allowed to react at room temperature for 0.5 hour. After completion of the reaction, the solvent was removed by distillation under reduced pressure. The crude product was purified by silica gel column chromatography [dichloromethane / methanol = 82:18 (v / v)] to afford the title compound 7 (76 mg, yield: 100%). ESI-MS m / z: 612 (M+H).

[0072] Synthesis of Compound 9: Compound 7 (76 mg, 0.124 mmol) and commercially available compound 8 (46 mg, 0.149 mmol) were mixed in 3 mL of N,N-dimethylformamide. N,N-diisopropylethylamine (32 mg, 0.248 mmol) was added and the mixture was allowed to react at room temperature for 2 hours. After completion of the reaction, the solvent was removed by distillation under reduced pressure. The crude product was purified by silica gel column chromatography [dichloromethane / methanol = 82:18 (v / v)] to afford the title compound 9 (70 mg, yield: 70%). ESI-MS m / z: 805 (M+H).

[0073] Synthesis of Compound 10: Compound 9 (70 mg, 0.087 mmol) was dissolved in 3 mL of N,N-dimethylformamide, and Bis-PNP (40 mg, 0.130 mmol) and N,N-diisopropylethylamine (22 mg, 0.174 mmol) were added sequentially. The mixture was allowed to react at room temperature for 16 hours. After completion of the reaction, the solvent was removed by distillation under reduced pressure. The crude product was purified by silica gel column chromatography [dichloromethane / methanol = 90:10 (v / v)] to afford the title compound 10 (40 mg, yield: 47.6%). ESI-MS m / z: 970 (M+H).

[0074] Synthesis of compound LK-322022: Compound 10 (14 mg, 0.014 mmol) was dissolved in 1 mL of N,N-dimethylformamide, and N,N-diisopropylethylamine (3.5 mg, 0.027 mmol) and commercially available eribulin (5 mg, 0.007 mmol) were added sequentially. The mixture was allowed to react at room temperature for 16 hours. After completion of the reaction, the reaction was monitored by liquid chromatography-mass spectrometry. After addition of 5 mL of water, the mixture was extracted three times with 10 mL of ethyl acetate. The organic phases were separated, combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by preparative liquid chromatography to afford LK-322022 (5.1 mg, yield: 48.1%). ESI-MS m / z: 1560 (MH).

[0075] Example 3 Preparation of Conjugate 7B7Mc-VAGGFG-PAB-Eribulin (ILB-3101)

[0076] Take the antibody 7B7 to be conjugated in Example 1 above, take 3 mg of the antibody, dilute it to 3 mg / mL with 10 mM sodium phosphate buffer, pH 7.4, and add tris(2-hydroxyethyl)phosphine hydrochloride (TCEP) stock solution. The TCEP stock solution is prepared by preparing a 10 mM stock solution in 10 mM sodium phosphate buffer, pH 7.4 (TCEP, Sigma, Cat. No. C4706-2G, CAS 51805-45-9). Ensure that the final molar concentration of TCEP to the final molar concentration of antibody is 4:1. The antibody is reduced at 37°C for 45 minutes. Add the linker-eribulin stock solution in Example 2. The stock solution is prepared by dissolving the linker-eribulin to 5 mM in dimethyl sulfoxide (DMSO). Ensure that the final molar concentration of the linker-eribulin to the final molar concentration of the antibody is 6:1. The antibody is conjugated at 2-8°C for 2 hours. The antibody was then transferred to a 30KD ultrafiltration tube (Merck, catalog number UFC9030), placed at 3000g centrifugal force at 2-8°C for liquid exchange to reduce the residual toxin to one thousandth of the reaction concentration, followed by sterile filtration, determination of concentration, and aliquoting for cryopreservation.

[0077] To obtain products with different conjugation ratios (DARs), the molar ratio of antibody:reducing agent:linker toxin was adjusted between 1:1 and 6:2 and 15, while the other conditions remained unchanged. The DAR value of the final conjugate ILB-3101 was determined based on the test results.

[0078] Example 4 DAR value detection and purity detection of conjugate ILB-3101

[0079] Coupling ratio detection analysis

[0080] The average drug-adapted average (DAR), i.e., the conjugation ratio, was determined by hydrophobic chromatography-HPLC using a TSKgel Butyl-NPR column, 4.6 mm × 10 cm, 2.5 μm, manufactured by TOSOH, Cat. No. 042168. Approximately 0.3 mg of the sample to be tested was added to 150 μL of mobile phase A, and the mixture was centrifuged at 10,000 g for 5 minutes. The supernatant was collected. Mobile phase A consisted of 20 mM sodium phosphate + 1.5 M ammonium sulfate, pH 7.0; mobile phase B consisted of 20 mM sodium phosphate + 20% (v / v) acetonitrile, pH 7.0. The analytical column was connected to an Agilent 1260 HPLC at a flow rate of 0.6 mL / min. Mobile phase A was used for 30 minutes or more until the UV baseline at 280 nm stabilized. A 50 μg sample was injected. After a 2-minute mobile phase A wash, an 18-minute gradient elution (0% B–100% B) was performed. The wash was continued at 100% mobile phase B for 5 minutes before elution. The percentage of each DAR value was calculated based on its peak area. The product of the peak percentage and the DAR value was summed and divided by the total percentage to obtain the average DAR value.

[0081] The average DAR value of the conjugate ILB-3101 was 3.5.

[0082] Purity detection and analysis

[0083] Purity was determined by SEC-HPLC using a TSKgel G3000SWXL molecular sieve separation method. The analytical column was TOSOH, catalog number 08541, 7.8 mm × 30 cm, 5 μm. Approximately 0.3 mg of the sample was centrifuged at 10,000 g for 5 minutes in a volume of 300 μL. The supernatant was collected. The mobile phase consisted of 50 mM sodium phosphate and 0.1 M sodium chloride, pH 6.8. The column was connected to an Agilent 1260 HPLC instrument at a flow rate of 0.8 mL / min. The column was flushed with the mobile phase for at least 30 minutes until the UV baseline at 280 nm stabilized. A 100 μg sample was injected and the mobile phase was flushed for 20 minutes. The percentages of the high-molecular-weight, monomeric, and low-molecular-weight peaks were calculated based on the peak areas.

[0084] The SEC-HPLC purity of the conjugate ILB-3101 was 95.88%.

[0085] Example 5 Comparison of in vitro tumor inhibition effects of conjugate ILB-3101

[0086] Cell viability assay

[0087] Human Calu-6 (Procell, Catalog No. CL-327, human degenerative lung cancer cells) were revived and seeded into 96-well culture plates in DMEM / F12 (Hyclone, Catalog No. SH30023.01) containing 10% FBS (Gibco, Catalog No. 10099141). 2500 cells per well were cultured in 150 μl of culture medium at 37°C in a 5% carbon dioxide incubator. After about 20 hours, the cells were incubated with the conjugate ILB-3101 and the antibody-drug conjugate DS- 7300a (Daiichi Sankyo Co., Ltd.) culture medium was added to 50 μl of 12 dilutions with a final drug concentration between 0 and 100 nM (100 nM concentration in the first well, followed by 10 5-fold dilutions, with the final drug at zero, and repeated for each sample well). After continuing co-culture for 4 to 7 days, the culture plate was removed and 50 μl of CTG detection reagent (CellTiterGlo, Promega, Cat. No. G7575) was added to each well. The reaction was carried out for 2 minutes according to the recommended operation of the kit, and the fluorescence value was measured using Tecan Spark. The average value of the replicates was taken as the Y-axis, and the log10 value of the dilution gradient was used as the X-axis to make a smooth curve graph. The EC / IC was calculated according to the four parameters. 50 value.

[0088] A similar cell activity assay was used, but the cell lines were replaced with A673 human rhabdomyosarcoma cells, HT1080 human fibrosarcoma cells, PA1 human ovarian teratoma cells, OVCAR3 human ovarian cancer cells, PC9 non-small cell lung cancer cells, MDA-MB-468 HER-2 positive triple-negative breast cancer cells, and SK-BR-3 human triple-negative breast cancer cells for tumor inhibition testing. The experimental results, as shown in Figures 1 and 2, show that the conjugate ILB-3101 exhibited significant inhibitory effects on all tumor cell lines. Compared to DS-7300a, ILB-3101 exhibited superior activity against multiple tumor cell lines, with IC 50 About 10-100pM.

[0089] Example 6 In vivo pharmacodynamic analysis of the coupled product ILB-3101

[0090] Bioduro (www.bioduro-sundia.com) was commissioned to conduct an in vivo efficacy evaluation. A well-cultured human breast cancer cell line (MDA-MB-468) was inoculated into immunodeficient mice (B-NDG) at a rate of 10 million cells per mouse until the tumor grew to 100 mm. 3After randomization, the next day, ILB-3101 was administered once to three groups at 1.5 mg / kg, 3 mg / kg, and 6 mg / kg, while DS-7300a was administered to two groups at 3 mg / kg and 6 mg / kg, respectively. Tumor size was measured twice weekly until the end of the experiment. The entire procedure was approved by the Experimental Animal Research Committee.

[0091] Using the same animal model, the cell line was replaced with the OVCAR3 ovarian cancer cell line. ILB-3101 was administered to the three groups at 1.5 mg / kg, 3 mg / kg, and 6 mg / kg, while DS-7300 was administered to the three groups at 3 mg / kg, 6 mg / kg, and 12 mg / kg, respectively. Tumor size was measured twice weekly until the end of the experiment. The entire procedure was approved by the Experimental Animal Care Committee.

[0092] The same animal model was used, but the cell line was replaced with the SK-OV-3 ovarian cancer cell line. ILB-3101 was administered to the two groups at 3 mg / kg and 6 mg / kg, while DS-7300 was administered to the two groups at 3 mg / kg and 6 mg / kg, respectively. Tumor size was measured twice weekly until the end of the experiment. The entire procedure was approved by the Experimental Animal Care Committee.

[0093] The same animal model was used, but the cell line was replaced with the A2780 ovarian cancer cell line. ILB-3101 was administered to the two groups at 3 mg / kg and 6 mg / kg, while DS-7300 was administered to the two groups at 3 mg / kg and 6 mg / kg, respectively. Tumor size was measured twice weekly until the end of the experiment. The entire procedure was approved by the Experimental Animal Care Committee.

[0094] Using the same animal model, the cell line was replaced with the JIMT-1 HER-2-positive breast cancer cell line. ILB-3101 was administered to the two groups at 3 mg / kg and 6 mg / kg, while DS-7300 was administered to the two groups at 3 mg / kg and 6 mg / kg, respectively. Tumor size was measured twice weekly until the end of the experiment. The entire procedure was approved by the Experimental Animal Care Committee.

[0095] The same animal model was used, but the cell line was replaced with PC-9 non-small cell lung cancer cells. ILB-3101 was administered to the two groups at 3 mg / kg and 6 mg / kg, and DS-7300 was administered to the two groups at 3 mg / kg and 6 mg / kg, respectively, for a cumulative dose of one. Tumor size was measured twice weekly until the end of the experiment. The entire procedure was approved by the Experimental Animal Research Committee.

[0096] Using the same animal model, the U937 acute myeloid leukemia cell line was replaced with the ILB-3101 (0.15 mg / kg, 0.75 mg / kg, 3 mg / kg, and 6 mg / kg) cell line. Eribulin (0.15 mg / kg) and isotype ADC eribulin (6 mg / kg) were administered once for all test groups and the control group on day 7 after tumor cell inoculation. On day 20, the ILB-3101 (6 mg / kg) and isotype ADC eribulin (6 mg / kg) groups received an additional dose. Tumor cell fluorescence intensity, which reflects tumor cell number, was measured once or twice weekly until the end of the experiment. The entire procedure was approved by the Experimental Animal Care Committee. This demonstrates that ILB-3101 exhibits potent tumor suppressive activity in this acute myeloid leukemia animal model.

[0097] Using the same animal model, the cell line was replaced with the LK-2 squamous cell lung carcinoma cell line. ILB-3101 was administered at 3 and 6 mg / kg, and DS-7300 at 6 mg / kg. Dosing was repeated twice, on days 6 and 20 after tumor cell inoculation. Tumor size was measured twice weekly until the end of the experiment. The entire procedure was approved by the Experimental Animal Care Committee. This demonstrates that ILB-3101 exhibits potent tumor suppressive activity in this human squamous cell lung carcinoma model.

[0098] The experimental results, shown in Figures 3-6, show that the conjugate ILB-3101 significantly inhibited tumor volume in multiple tumor cell lines. Compared to DS-7300a, ILB-3101 demonstrated superior or equivalent in vivo anti-tumor activity. ILB-3101 also demonstrated significant anti-tumor activity in a DS7300a-resistant xenograft model.

[0099] Example 7 In vivo toxicity and pharmacokinetic testing of the conjugated product ILB-3101

[0100] The conjugate, ILB-3101, was administered intravenously to cynomolgus monkeys (2 monkeys per sex) every 3 weeks for 2 consecutive doses to test for potential toxicity.

[0101] experimental animals

[0102] Animal level: ordinary, non-naive;

[0103] Age at start of dosing (D1): Males: 3.7 years; Females: 3.0 years;

[0104] Body weight before drug administration (D-3): male: 3.10 kg, female: 3.10 kg;

[0105] Source of experimental animals: Guangxi Xiongsen Primate Experimental Animal Breeding and Development Co., Ltd.

[0106] Laboratory Animal Production License Number: SCXK(Gui)2021-0004;

[0107] Laboratory animal quality certificate number: 0002975 (male), 0002990 (female);

[0108] Production license issuing unit: Guangxi Zhuang Autonomous Region Science and Technology Department.

[0109] ILB-3101 was administered at 4 mg / kg and 8 mg / kg on Day 1 and Day 22, respectively. Animals were administered via intravenous infusion at a volume of 5 mL / kg. The day of first administration was defined as Day 1. Surviving animals were autopsied on Day 36.

[0110] During the experiment, the animals were subjected to clinical observation, body weight, food intake, body temperature, electrocardiogram, clinical pathology (blood cell count, coagulation function, blood biochemistry), immune cell phenotype, gross anatomy and histopathology and toxicokinetic analysis.

[0111] result

[0112] Death / Moribundity: During the study, no test article-related death or moribundity was observed in animals in each dose group.

[0113] At the 4 mg / kg dose (administered on D1): Male animals experienced significant decreases in food intake on D1, D9, and D11, while female animals experienced significant decreases on D1, D9, and D10. WBC, Neutral, Lymph, and Mono levels decreased most significantly on D8 (one week after administration) in both males and females, with recovery observed on D21. Eos decreased on D8 and D21 in males and on D3, D8, and D21 in females. RBC and HGB levels decreased on D21 (three weeks after administration). Retic levels decreased on D8 (one week after administration), with recovery or a recovery trend observed on D21. PLT levels decreased on D3 and D21. CD3+, CD4+, and CD8+ T lymphocyte counts in both males and females began to decrease on D3 or D8, respectively, two days and one week after the first dose, with recovery observed on D21.

[0114] At the 8 mg / kg dose (administered on D22), food intake continued to decrease significantly in male animals on D22, D26, D27, D31, and D32, and in female animals on D29 and D31. Compared to the first dose, WBC, Neut, Lymph, and Mono levels in both males and females decreased most significantly on D29 (one week after administration), with even greater decreases after the second, higher dose (8 mg / kg). Significant increases were observed in WBC, Neut, Lymph, Mono, Eos, and Baso levels on D36 after the final dose. RBC and HGB levels continued to decrease from D24 to D36, with a more pronounced decrease compared to the first dose. Retic levels in males continued to decrease on D29 (one week after administration), with recovery observed on D36. PLT levels continued to decrease on D24, with recovery observed on D36. The CD4+ / CD8+ ratio increased in both male and female animals on Day 24. CD3+, CD4+, and CD8+ T lymphocyte counts decreased similarly to those observed after the first dose on Day 24 or Day 29, but recovered by Day 36. FIB increased on Day 36, as did TBil on Day 24, in both male and female animals. Microscopic observation of the thymic cortex and medulla of euthanized animals revealed a mild decrease in lymphocytes.

[0115] During the trial, no abnormalities related to the test article were found in the clinical observation, body temperature, electrocardiogram, and gross anatomical examination of male and female animals.

[0116] Toxicokinetics: Serum concentrations of ILB-3101 and ILB-3101 total antibody were determined by ELISA, with a limit of quantification of 10.0 ng / mL. Serum concentrations of eribulin were determined by LC-MS / MS, with a limit of quantification of 145.98 pg / mL (0.200 nmol / L). Toxicokinetic parameters were calculated using the non-compartmental model (NCA) using WinnonLin 8.0.0.3176.

[0117] The toxicokinetic parameters of ILB-3101 and ILB-3101 total antibody in each group of animals are as follows:

[0118] After repeated intravenous infusion of ILB-3101 at doses of 4 mg / kg and 8 mg / kg to crab-eating monkeys, the peak drug concentration (Cmax) and drug exposure (AUC) of ILB-3101 in the animals were lower than the total antibody, and less eribulin existed in the serum in free form.

[0119] Table 1 The toxicokinetic parameters of ILB-3101 and ILB-3101 total antibody in each group of animals are as follows:

[0120] On day 22, approximately 1.2 mL of blood was collected from the subcutaneous vein of the hind limb. Blood was collected immediately (±1 min) after the end of dosing on day 22, and at 1 hour, 4 hours, 8 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 168 hours, 336 hours, and 504 hours after the start of dosing. The plasma concentration curve of ILB-3101 (8 mg / kg) in cynomolgus monkeys on day 22 is shown in Figure 7.

[0121] Conclusion: In summary, ILB-3101 was administered intravenously to cynomolgus macaques at doses of 4 mg / kg and 8 mg / kg on D1 and D22, respectively. No animals were found to be dying or near-dying. The main toxic reactions of ILB-3101 after administration were: decreased appetite, immune-related toxicity (including decreased WBC, Neut, Lymph, Mono, and Eos, and mild lymphocytopenia in the thymic cortex / medullary cells), and effects on red blood cell-related indices (decreased RBC, HGB, and Retic). Under the conditions of this study, the highest non-severely toxic dose (HNSTD) was 8 mg / kg. At this dose, the AUClast of ILB-3101 in animals was 2.62 h*mg / mL, and the Cmax was 148 μg / mL. The AUClast of ILB-3101 total antibody was 10.8 h*mg / mL, and the Cmax was 184 μg / mL. The AUClast of the animal small molecule toxin Eribulin was 199 h*ng / mL, and the Cmax was 2720 pg / mL.

[0122] It can be seen that the conjugate ILB-3101 is well tolerated in crab-eating macaques, with HNSTD ≥ 6 mg / kg and maximum tolerated dose MTD ≥ 8 mg / kg, indicating that the conjugate ILB-3101 has a wider therapeutic window and better application prospects.

[0123] The above-described embodiments merely represent several implementation methods of the present application. While the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the patent. It should be noted that a person skilled in the art could make several modifications and improvements without departing from the spirit of the present application, all of which fall within the scope of protection of the present application. Therefore, the scope of protection of the patent in this application shall be based on the appended claims, and the specification and drawings may be used to interpret the claims.

Claims

1. An anti-B7H3 antibody-eribulin conjugate, or a pharmaceutically acceptable salt or solvate thereof, wherein the structure of the anti-B7H3 antibody-eribulin conjugate is as follows: in, TL is an anti-B7H3 antibody, and n is an integer or decimal from 1 to 20.

2. The anti-B7H3 antibody-eribulin conjugate, or a pharmaceutically acceptable salt or solvate thereof according to claim 1, wherein: n is an integer or decimal from 3 to 5.

3. The anti-B7H3 antibody-eribulin conjugate, or a pharmaceutically acceptable salt or solvate thereof according to claim 1 or 2, wherein: The heavy chain variable region HCVR of the anti-B7H3 antibody is shown in SEQ ID NO: 1, the light chain variable region LCVR is shown in SEQ ID NO: 2, the heavy chain constant region is shown in SEQ ID NO: 3, and the light chain constant region is shown in SEQ ID NO:

4. 4 . A pharmaceutical composition comprising the anti-B7H3 antibody-eribulin conjugate according to claim 1 , a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable excipient, diluent or carrier. The pharmaceutical composition according to claim 4 , further comprising one or more other active ingredients.

6. Use of the anti-B7H3 antibody-eribulin conjugate according to any one of claims 1 to 3, a pharmaceutically acceptable salt or solvate thereof, or the pharmaceutical composition according to any one of claims 4 to 5 in the preparation of a medicament for treating and / or preventing a B7H3-mediated disease or condition.

7. The use according to claim 6, wherein The disease or disorder is a disease or disorder in which B7H3 is positively expressed.

8. The use according to claim 6 or 7, wherein The disease or condition is cancer.

9. The use according to claim 8, wherein The cancer is selected from breast cancer, ovarian cancer, ovarian teratoma, cervical cancer, uterine cancer, prostate cancer, kidney cancer, urethral cancer, testicular cancer, bladder cancer, liver cancer, gastric cancer, head and neck cancer, endometrial cancer, salivary gland cancer, esophageal cancer, esophageal cancer, lung cancer, colon cancer, rectal cancer, colorectal cancer, bone cancer, skin cancer, thyroid cancer, pancreatic cancer, melanoma, neural tumor, glioma, sarcoma, squamous cell carcinoma, lymphoma, myeloma and leukemia. Preferably, the cancer is selected from breast cancer, ovarian cancer, ovarian teratoma, lung cancer, sarcoma and leukemia.

10. The use according to claim 9, wherein the cancer is selected from triple-negative breast cancer, HER-2 positive breast cancer, luminal A breast cancer, luminal B breast cancer, rhabdomyosarcoma, fibrosarcoma, chronic leukemia, acute leukemia, metastatic castration-resistant prostate cancer, non-small cell lung cancer, small cell lung cancer, degenerative lung cancer, ovarian epithelial tumor, ovarian sex cord-stromal tumor, ovarian germ cell tumor, ovarian metastatic tumor, neuroblastoma, glioma, glioblastoma multiforme, head and neck squamous cell carcinoma, non-Hodgkin's lymphoma, diffuse large B-cell lymphoma, multiple myeloma; preferably, the cancer is selected from triple-negative breast cancer, HER-2 positive breast cancer, rhabdomyosarcoma, fibrosarcoma, non-small cell lung cancer, degenerative lung cancer, and acute leukemia.

11. The use according to claim 9, wherein the cancer is selected from HER-2 positive triple-negative breast cancer, glioblastoma, chronic lymphocytic leukemia, acute myeloid leukemia, acute lymphocytic leukemia, chronic myeloid leukemia, chronic myeloid leukemia, embryonal rhabdomyosarcoma, alveolar rhabdomyosarcoma, pleomorphic rhabdomyosarcoma, sclerosing rhabdomyosarcoma, and squamous cell carcinoma of the lung; preferably, the cancer is selected from HER-2 positive triple-negative breast cancer, acute myeloid leukemia, and squamous cell carcinoma of the lung.

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