Pharmaceutical composition and use thereof

Abstract

The invention belongs to the field of biological medicine, and relates to a pharmaceutical composition and application thereof. Specifically, the pharmaceutical composition comprises a first active component and a second active component, and the first active component is an antibody or an antigen binding fragment thereof; the second active component is an antibody-conjugated drug, and the antibody-conjugated drug comprises the antibody or the antigen-binding fragment thereof, a linker and an effective load. The pharmaceutical composition disclosed by the invention has a good anti-tumor effect and relatively high safety.

Description

[0001] Cross Reference to Related Applications

[0002] This application is based on and claims priority to the application with CN application number 202410561279.4, filed on May 7, 2024, the entire contents of which are incorporated herein by reference in its entirety. TECHNICAL FIELD

[0003] The present application belongs to the field of biological medicine, and relates to a drug combination and use thereof. Specifically, the drug combination comprises an antibody and an antibody conjugate drug of the antibody. More specifically, the antibody is an anti-HER3 antibody, and the antibody conjugate drug is an antibody conjugate drug of the anti-HER3 antibody; or the antibody is an anti-TROP2 antibody, and the antibody conjugate drug is an antibody conjugate drug of the anti-TROP2 antibody. The present application also relates to a pharmaceutical composition comprising the drug combination and use thereof. BACKGROUND

[0004] Antibody-drug conjugate (ADC) is a new type of macromolecular targeted drug, which has the strong killing effect of traditional small molecule chemotherapy and the tumor targeting of antibody drugs. ADC is composed of three main parts: antibody responsible for selective recognition of cancer cell surface antigens, payload responsible for killing cancer cells, and linker connecting antibody and payload. After ADC drugs enter the blood circulation, their antibody parts specifically bind to the antigens on the surface of tumor cells, and the complex formed by the combination enters the cell through receptor-mediated endocytosis (which can be divided into four types: clathrin-mediated endocytosis, cave-like invagination, macropinocytosis, and clathrin- and caveolin-independent endocytosis). The cleavable linker is sensitive to environmental factors in tumor cells and will be cleaved by specific pH environment, proteases, or certain chemicals; ADC drugs carrying non-cleavable linkers are digested by lysosomes, releasing the drugs. Some small molecules of ADC drugs can penetrate the cell membrane to further kill surrounding cancer cells, i.e., bystander killing effect (Khongorzul, Puregmaa, et al. "Antibody-Drug Conjugates: A Comprehensive Review." Mol Cancer Res 1 (2020).; Kostova Vesela, Désos Patrice, Starck et al. The Chemistry Behind ADCs. [J]. Pharmaceuticals (Basel), 2021, 14: undefined.).

[0005] The accumulation of the payload in the tumor is limited, so it is necessary to kill the tumor at sub-nanomolar concentrations. In addition, it is also necessary to have good stability to maintain the structure and activity in the circulatory system and lysosomes. Common payloads mainly include microtubulin inhibitors and DNA damage agents. Microtubulin inhibitors can kill tumor cells and inhibit the rapid proliferation of tumor cells by inhibiting the generation and aggregation of microtubulin. The microtubulin inhibitors commonly used for ADC construction include Maytansines, Auristatins and Tubulysins, etc. DNA damage agents are divided into three categories according to different mechanisms: DNA double-strand breakage agents, DNA insertion agents and DNA alkylation agents. DNA is crucial for cell growth and proliferation, and its destruction can efficiently kill tumor cells and inhibit their rapid proliferation. The DNA damage agents commonly used for ADC construction include pyrrolobenzodiazepine and indolinobenzodiazepine (such as pyrrolo[2,1-c][1,4]benzodiazepine, PBD), duocarmycin, camptothecin (CPT) and its derivatives (such as topoisomerase I inhibitor DXd and SN-38), and calicheamicin, etc. (Dan Nirnoy, Setua Saini, Kashyap Vivek K et al. Antibody-Drug Conjugates for Cancer Therapy: Chemistry to Clinical Implications. [J]. Pharmaceuticals (Basel), 2018, 11: undefined.).

[0006] ADC has a broad application prospect, but such anti-cancer drugs are hindered by a relatively narrow therapeutic window. More potent payloads can reduce the minimum effective dose (MED) of ADC, but they often cause greater toxicity, resulting in a lower maximum tolerated dose (MTD). Their clinical success is often limited by low tumor penetration and poor distribution at relevant doses. The tumor distribution of ADC depends on many factors, including the dose itself, the physicochemical properties of the linker-payload, the systemic clearance rate, the amount of antigen expression, the endocytosis rate, and the tumor's surface-to-volume ratio (S / V), among others. Poor tissue penetration of ADC drugs is a major challenge in solid tumor antibody therapy. For ADC, on the one hand, the dose-limiting toxicity of small molecule payloads results in lower dosing than the parent antibody, reducing tumor penetration. Even with continuous dosing, the sustained internalization of ADC prevents delivery to all tumor cells. On the other hand, high antigen expression on solid tumors and high affinity between antigen and antibody result in most ADC binding to antigen being limited to the vicinity of tumor blood vessels, forming a binding-site-barrier (BSB) that prevents penetration into tumor cells away from blood vessels. There is little payload in deep tumor cells, resulting in a decrease in the tumor clearance efficiency of ADC drugs. For ADC drugs that do not exhibit bystander killing effects, the binding-site-barrier effect is more pronounced.

[0007] HER3 is a member of the human epidermal receptor (HER) family, consisting of an extracellular ligand-binding region, an alpha-helix transmembrane region, and an intracellular tyrosine kinase region. HER3 is abnormally overexpressed in various invasive tumors, including breast tumors, non-small cell lung cancer (NSCLC), metastatic colon cancer, head and neck cancer, pancreatic cancer, ovarian cancer, clear cell sarcoma, gastric cancer, and skin cancer. HER3 can bind to the ligand neuregulin (NRG), although it lacks intrinsic tyrosine kinase activity, but HER3 can induce cross-linking phosphorylation of highly conserved kinase residues in the cell by forming heterodimers with HER2, i.e., one receptor phosphorylates specific tyrosine residues on another receptor, thereby recruiting and activating downstream proteins, directly activating the PI3K / AKT pathway, causing signal cascade reactions, and promoting tumor cell development. Therefore, HER3 has become a new target for tumor therapy.

[0008] Trop2 promotes tumor growth and metastasis mainly by regulating multiple cell signaling pathways. After the intracellular tail serine residue (S303) of Trop2 is phosphorylated, it promotes the hydrolysis of 4,5-diphospho phosphatidylinositol (PIP2) into inositol triphosphate (IP3) and diacylglycerol (DAG), which in turn releases calcium ions from the endoplasmic reticulum, activates the MAPK signaling pathway, and promotes cell cycle progression [Wang J. Research Progress of Trop2 Gene and Triple-negative Breast Cancer [J]. Surgical Theory and Practice, 2022. DOI: 10.16139 / j.1007-9610.2022.05.018.]. Overexpression of Trop2 can also increase p42 / p44MAPK (ERK1 / 2) phosphorylation, thereby further enhancing the activity of downstream target AP-1 transcription factor, down-regulating Bcl-2 expression, and inhibiting apoptosis. In addition, AP-1 can also increase the expression of cyclin D1, cyclin E, and cyclin-dependent kinase (CDKs), inhibit the formation of cyclinD1-CDK4 and cyclinE-CDK4 complexes, and promote the transition of the cell cycle from G0 / G1 to S phase. At the same time, Trop2 can up-regulate the expression of the proliferation marker Ki-67, thereby promoting tumor cell proliferation [Liu T, Liu Y, Bao X, et al. Overexpression of TROP2 predicts poor prognosis of patients with cervical cancer and promotes the proliferation and invasion of cervical cancer cells by regulating ERK signaling pathway [J]. PLoS One, 2013, 8(9): e75864.]. In addition, Trop2 can promote the enrichment of cytoplasmic protein kinase C receptor 1 (RACK1) on the cell membrane, making it close to integrin β-1, reducing the binding of fibronectin to integrin β-1, and forming a complex of Trop2, integrin β-1, and talin protein, which together enhances the function of focal adhesion kinase FAK, thereby reducing tumor cell adhesion and promoting metastasis and diffusion [Yu Q, Miao QF. Research Progress of Anti-tumor Drug Target Trop2 [J]. Chinese Medicine and Biotechnology, 2018, 13(4): 5. DOI: 10.3969 / j.issn.1673-713X.2018.04.012.].

[0009] In summary, the development of ADC drugs with significant efficacy and small side effects and their administration strategies to further improve the efficacy and safety of ADCs has broad prospects. SUMMARY

[0010] The present inventors have made intensive studies and creative efforts, and as a result, have completed a pharmaceutical combination which has a good effect of treating or preventing a tumor or an autoimmune disease, and / or a high safety. The present invention has been made on the basis of the above findings.

[0011] One aspect of the present invention relates to a pharmaceutical combination comprising a first active ingredient and a second active ingredient, wherein:

[0012] The first active ingredient is an antibody or an antigen-binding fragment thereof;

[0013] The second active ingredient is an antibody-drug conjugate comprising the antibody or the antigen-binding fragment thereof, a linker, and a payload.

[0014] In the pharmaceutical combination of the present invention, the antibody or the antigen-binding fragment thereof comprised in the antibody-drug conjugate as the second active ingredient is the same as the antibody or the antigen-binding fragment thereof as the first active ingredient.

[0015] In some embodiments of the present invention, the pharmaceutical combination comprises an antibody and an antibody-drug conjugate of the antibody.

[0016] In some embodiments of the present invention, the pharmaceutical combination consists of an antibody and an antibody-drug conjugate of the antibody.

[0017] In some embodiments of the present invention, the pharmaceutical combination consists of active ingredients of an antibody and an antibody-drug conjugate of the antibody.

[0018] In some embodiments of the present invention, the pharmaceutical combination consists of a first active ingredient and a second active ingredient.

[0019] In some embodiments of the present invention, the pharmaceutical combination consists of active ingredients of a first active ingredient and a second active ingredient.

[0020] In some embodiments of the present invention, the pharmaceutical combination, wherein the antibody is a mono-specific antibody, a bi-specific antibody, or a multi-specific antibody (e.g., a tri-specific antibody).

[0021] In some embodiments of the present invention, the pharmaceutical combination, wherein the antibody is a human IgG antibody, preferably a human IgG1 antibody or a human IgG4 antibody.

[0022] In some embodiments of the present invention, the pharmaceutical combination, wherein the antibody is an antibody for treating a tumor or an autoimmune disease.

[0023] Suitable antibodies can bind to any disease-related antigen known in the art. When used against tumors, for example, many antigens expressed by or otherwise associated with tumor cells are known in the art, including but not limited to carbonic anhydrase IX, alpha-fetoprotein (AFP), alpha-actinin-4, A3, ART-4, B7, Ba 733, BAGE, BrE3 antigen, CA125, CAMEL, CAP-1, CASP-8 / m, CCL19, CCL21, CD1, CD1a, CD2, CD3, CD4, CD5, CD8, CD11A, CD14, CD15, CD16, CD18, CD19, CD20, CD21, CD22, CD23, CD25, CD29, CD30, CD32b, CD33, CD37, CD38, CD40, CD40L, CD44, CD45, CD46, CD52, CD54, CD55, CD59, CD64, CD66a-e, CD67, CD70, CD70L, CD74, CD79a, CD80, CD83, CD95, CD126, CD132, CD133, CD138, CD147, CD154, CDC27, CDK-4 / m, CDKN2A, CTLA-4, CXCR4, CXCR7, CXCL12, colon-specific antigen p (CSAp), CEA (CEACAM5), CEACAM6, c-Met, EGFR, EGFRvIII, Trop-2, EGP-2, Ep-CAM, her2, her3, claudin 18.2, ROR1, ROR2, dll3, folate receptor, G250 antigen, HLA-DR, HM1.24. Human chorionic gonadotropin (HCG) and its subunits, HMGB-1, IGF-1R, IFNRs, IL-4R, IL-6R, IL-13R, IL-15R, IL-17R, IL-18R, IL-2R, IL-8R, IL-12R, IL-15R, IL-17R, IL-18R, IL-23R, IL-25R, Insulin-like growth factor 1 (IGF-l), IGF-1R, MAGE, MAGE-3, Mucins (MUC) and their receptors, MUM-1 / 2, MUM-3, NCA66, NCA95, NCA90, PD-1, PD-L1, VEGFRs, PRAME, PSMA, ILGF-1R, T101, TAG-72, TRAIL receptors, Tumor necrosis factor, VEGNR, ED-B fibronectin, 17-1A antigen, Complement factors C3, C3a, C3b, C5a, C5, bd-6, Kras, oncogene markers and oncogene products (see, e.g., Sensi et al. Clin Cancer Res 2006 12:5023-32; Parmiani J Immunol 2007, 178:1975-79; Novellino et al. Cancer Immunol Immunother 2005, 54: 187-207).

[0024] In some embodiments of the application, the pharmaceutical combination is sought, wherein the antibody is an antibody targeting a tumor-associated antigen and / or an antibody targeting an immune checkpoint, or an antibody targeting an autoimmune disease-associated target;

[0025] Preferably, the tumor-associated antigen is one or more selected from the group consisting of PD-L1, CD19, CD20, CD22, CD30, CD33, CD79b, EGFR, Claudin 18.2, BCMA, HER2, HER3, TROP2, ROR1, CEACAM5, MET, Tissue Factor (TF), Folate Receptor alpha (FRa), NaPi2b, and Nectin-4;

[0026] Preferably, the immune checkpoint is one or more selected from the group consisting of PD-1, PD-L1, CTLA-4, LAG3, TIGIT, LMTK3, IDO, and TIM-3;

[0027] Preferably, the autoimmune disease-associated target is one or more selected from the group consisting of TNF-a, IL-1R, IL-2R, IL-6R, IL-7R, IL-17AR, IL-12R, IL-23R, and Integrin.

[0028] In some embodiments of the application, the pharmaceutical combination, wherein the antibody is selected from the group consisting of, but not limited to: abciximab (anti-glycoprotein Ilb / IIIa), alemtuzumab (anti-CD52), bevacizumab (anti-VEGF), cetuximab (anti-EGFR), gemtuzumab (anti-CD33), ibritumomab (anti-CD20), panitumumab (anti-EGFR), rituximab (anti-CD20), tositumomab (anti-CD20), trastuzumab (anti-ErbB2), lambrolizumab (anti-PD1), atezolizumab (anti-PD-L1), MEDI4736 (anti-PD-L1), nivolumab (anti-PD-1), ipilimumab (anti-CTLA-4), abagovomab (anti-CA-125), adecatumumab (anti-EpCAM), atlizumab (anti-IL-6 receptor), benralizumab (anti-CD125), obinutuzumab, CC49 (anti-TAG-72), AB-PG1-XG1-026 (anti-PSMA), D2 / B (anti-PSMA), tocilizumab (anti-IL-6 receptor), basiliximab (anti-CD25), daclizumab (anti-CD25), efalizumab (anti-CD11a), muromonab-CD3 (anti-CD3 receptor), natalizumab (anti-alpha 4 integrin), omalizumab (anti-IgE); anti-TNF-alpha antibodies such as CDP571, MTNFAI, M2TNFAI, M3TNFAI, M3TNFABI, M302B, M303, infliximab, certolizumab pegol, anti-CD40L, adalimumab, Benlysta.

[0029] In some embodiments of the application, the pharmaceutical combination, wherein the antibody is an anti-HER3 antibody.

[0030] In some embodiments of the present application, the pharmaceutical combination, wherein the antibody comprises a heavy chain variable region and a light chain variable region,

[0031] wherein,

[0032] the heavy chain variable region comprises HCDR1 as set forth in SEQ ID NO: 1, HCDR2 as set forth in SEQ ID NO: 2, and HCDR3 as set forth in SEQ ID NO: 3;

[0033] the light chain variable region comprises LCDR1 as set forth in SEQ ID NO: 4, LCDR2 as set forth in SEQ ID NO: 5, and LCDR3 as set forth in SEQ ID NO: 6.

[0034] In some embodiments of the present application, the pharmaceutical combination, wherein the heavy chain variable region has an amino acid sequence as set forth in SEQ ID NO: 7, and the light chain variable region has an amino acid sequence as set forth in SEQ ID NO: 9.

[0035] In some embodiments of the present application, the pharmaceutical combination, wherein the antibody has a heavy chain constant region of Ig gamma-1 chain C region or Ig gamma-4 chain C region; and a light chain constant region of Ig kappa chain C region.

[0036] In some embodiments of the present application, the pharmaceutical combination, wherein the antibody has a heavy chain constant region with an amino acid sequence as set forth in SEQ ID NO: 11; and a light chain constant region with an amino acid sequence as set forth in SEQ ID NO: 13.

[0037] In some embodiments of the present application, the pharmaceutical combination, wherein the antibody is an anti-TROP2 antibody.

[0038] In some embodiments of the present application, the pharmaceutical combination, wherein the antibody comprises a heavy chain variable region and a light chain variable region,

[0039] wherein,

[0040] the heavy chain variable region comprises HCDR1 as set forth in SEQ ID NO: 21, HCDR2 as set forth in SEQ ID NO: 22, and HCDR3 as set forth in SEQ ID NO: 23;

[0041] the light chain variable region comprises LCDR1 as set forth in SEQ ID NO: 24, LCDR2 as set forth in SEQ ID NO: 25, and LCDR3 as set forth in SEQ ID NO: 26.

[0042] In some embodiments of the application, the pharmaceutical combination, wherein the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 17, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 18.

[0043] In some embodiments of the application, the pharmaceutical combination, wherein the heavy chain constant region of the antibody is Ig gamma-1 chain C region or Ig gamma-4 chain C region; and the light chain constant region is Ig kappa chain C region.

[0044] In some embodiments of the application, the pharmaceutical combination, wherein the amino acid sequence of the heavy chain constant region of the antibody is set forth in SEQ ID NO: 11; and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO: 13.

[0045] In some embodiments of the application, the pharmaceutical combination, wherein the amino acid sequence of the heavy chain constant region of the antibody is set forth in SEQ ID NO: 19; and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO: 13.

[0046] In some embodiments of the application, the pharmaceutical combination, wherein,

[0047] The antibody is a human IgGl antibody, and the heavy chain constant region thereof has one or more mutations selected from the group consisting of:

[0048] L234A and L235A;

[0049] L234A and G237A;

[0050] L235A and G237A;

[0051] or

[0052] L234A, L235A and G237A;

[0053] Preferably, the heavy chain constant region of the immunoglobulin further has one or more mutations selected from the group consisting of:

[0054] N297A, D265A, D270A, P238D, L328E, E233D, H268D, P271G, A330R, C226S, C229S, E233P, P331S, S267E, L328F, A330L, M252Y, S254T, T256E, N297Q, P238S, P238A, A327Q, A327G, P329A, K322A, T394D, G236R, G236A, L328R, A330S, P331S, H268A, E318A, and K320A.

[0055] In the present application, if not otherwise specified, the letter before the site represents the amino acid before mutation, and the letter after the site represents the amino acid after mutation.

[0056] In some embodiments of the present application, the pharmaceutical combination, wherein the antigen-binding fragment is selected from a Fab, a Fab', a F(ab')2, a Fd, a Fv, a dAb, a complementarity determining region fragment, a single-chain antibody, a humanized antibody, a chimeric antibody, or a diabody.

[0057] In some embodiments of the present application, the pharmaceutical combination, wherein the antigen-binding fragment is a VHH, a scFv, a Fv fragment, a Fab fragment, or a F(ab')2 fragment.

[0058] In some embodiments of the present application, the pharmaceutical combination, wherein,

[0059] The linker is a hydrazone bond, a disulfide bond, a thioether bond, or a peptide bond, or a chemical group comprising a hydrazone bond, a disulfide bond, a thioether bond, or a peptide bond;

[0060] Preferably, the linker is one or more selected from 6-maleimidocaproyl (MC), maleimidopropionyl (MP), valine-citrulline (Val-Cit), alanine-phenylalanine (Ala-Phe), alanine-alanine-alanine (Ala-Ala-Ala), Mc-Ala-Ala-Ala, p-aminobenzyloxycarbonyl (PAB), 6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (MC-Val-Cit-PAB), Mal-PEGn-Val-Cit-PAB (n is a positive integer selected from 1-20), Phe-Lys(Fmoc)-PAB, Aloc-D-Ala-Phe-Lys(Aloc)-PAB-PNP, Boc-Phe-(Alloc)Lys-PAB-PNP, and 3-(pyridine-2-yl disulfide) propionic acid perfluorophenyl ester.

[0061] In some embodiments of the present application, the pharmaceutical combination, wherein,

[0062] the effective payload is one or more selected from the group consisting of a tubulin inhibitor, a DNA damaging agent, a topoisomerase inhibitor, an ALK inhibitor, and a PARP inhibitor;

[0063] Preferably, the tubulin inhibitor is one or more selected from the group consisting of dolastatins, auristatin cytotoxic molecules, and maytansine cytotoxic molecules; preferably, the auristatin cytotoxic molecule is selected from the group consisting of monomethyl auristatin E (MMAE) and derivatives thereof, and monomethyl auristatin F (MMAF) and derivatives thereof; preferably, the maytansine cytotoxic molecule is selected from the group consisting of DM1 and derivatives thereof, and DM4 and derivatives thereof;

[0064] Preferably, the DNA damaging agent is one or more selected from the group consisting of calicheamicin, duocarmycin, and an anthramycin derivative (PBD);

[0065] Preferably, the topoisomerase inhibitor is selected from the group consisting of camptothecins and camptothecin derivatives; preferably, the camptothecin and camptothecin derivative is 7-ethyl-10-hydroxy camptothecin (SN-38) or DXd;

[0066] Preferably, the PARP inhibitor is Niraparib.

[0067] In some embodiments of the present application, the pharmaceutical combination, wherein the linker-payload is a compound of Formula 0 or a pharmaceutically acceptable salt or ester thereof;

[0068] the compound of Formula 0 comprises a compound of Formula III, n alanines, and a compound of Formula IV connected in that order,

[0069]

[0070] wherein,

[0071] n is 2, 3, 4, 5, or 6;

[0072] the compound of Formula III, the n alanines, and the compound of Formula IV are independently connected to each other two by two either directly (e.g., by a chemical bond) or through a chemical group.

[0073] Pharmaceutically acceptable salts of the compounds of the present application include conventional salts prepared from pharmaceutically acceptable inorganic or organic acids, or inorganic or organic bases. Examples of suitable acid addition salts include those formed with hydrochloric, hydrobromic, sulfuric, phosphoric, nitric, perchloric, fumaric, acetic, propionic, succinic, glycolic, formic, lactic, maleic, tartaric, citric, pamoic, malonic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, fumaric, toluenesulfonic, methanesulfonic, naphthalene-2-sulfonic, benzenesulfonic, hydroxynaphthoic, hydroiodic, malic, tannic, and the like. Examples of suitable base addition salts include those formed with sodium, lithium, potassium, magnesium, aluminum, calcium, zinc, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methylglucosamine, procaine and the like.

[0074] In some embodiments of the present application, the pharmaceutical combination, wherein the compound of Formula 0 is represented by Formula I,

[0075]

[0076] wherein,

[0077] A represents alanine;

[0078] m is 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12;

[0079] n is 2, 3, 4, 5, or 6.

[0080] In some embodiments of the present application, the compound of Formula I, wherein:

[0081] m is 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12, and n is 2.

[0082] In some embodiments of the present application, the compound of Formula I, wherein:

[0083] m is 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12, and n is 3.

[0084] In some embodiments of the present application, the compound of Formula I, wherein:

[0085] m is 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12, and n is 4.

[0086] In some embodiments of the present application, the compound of Formula I, wherein:

[0087] m is 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12, and n is 5.

[0088] In some embodiments of the application, the compound of formula I is:

[0089] m is 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12, and n is 6.

[0090] In some embodiments of the application, the pharmaceutical combination, wherein the compound of formula 0 is of formula II,

[0091]

[0092]

[0093] In some embodiments of the application, the pharmaceutical combination, wherein the payload is a tubulin inhibitor.

[0094] In some embodiments of the application, the pharmaceutical combination, wherein the payload is an auristatin class of cytotoxic molecules.

[0095] In some embodiments of the application, the pharmaceutical combination, wherein the payload is monomethyl auristatin E (MMAE) or a derivative thereof.

[0096] In some embodiments of the application, the pharmaceutical combination, wherein the payload is monomethyl auristatin F (MMAF) or a derivative thereof.

[0097] In some embodiments of the application, the pharmaceutical combination, wherein the linker is one or more selected from 6-maleimidocaproyl (MC), valine-citrulline (Val-Cit), and p-aminobenzyloxycarbonyl (PAB).

[0098] In some embodiments of the application, the pharmaceutical combination, wherein the linker is mc-vc-PAB.

[0099] In some embodiments of the application, the pharmaceutical combination, wherein the linker-payload is mc-vc-PAB-MMAE.

[0100] In some embodiments of the application, the pharmaceutical combination, wherein the antibody-drug conjugate, the linker is attached to the antibody or antigen-binding fragment thereof via one or more thioether bonds.

[0101] Preferably, the linker forms a thioether bond with the sulfur atom of the disulfide bond position of the hinge region of the antibody.

[0102] Without being bound by theory, the N-maleimide group alkylates with the thiol group in the antibody to form a stable thioether linkage.

[0103] In some embodiments of the present application, the pharmaceutical combination, wherein the average number of Linker-Payload conjugated to each antibody molecule (DAR) is 1-12, 1-11, 1-10, 1-9, 1-8, 2-8, 2-6, 2-4, 3-8, 4-8, 5-8, 6-8, 7-8, 1, 2, 3, 4, 5, 6, 7 or 8.

[0104] Those skilled in the art can understand that, since the average number is calculated, the specific value of the average number can be a positive integer or a non-integer such as a decimal number, and the number of digits after the decimal point of the decimal number is not particularly limited and can be 1 digit, 2 digits, 3 digits, etc. Taking the average number of 1-8 as an example: 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9 all fall within the range of 1-8. Similar understanding can also be made for the average number of 1-12, 1-11, 1-10, 1-9, 2-8, 2-6, 2-4, 3-8, 4-8, 5-8, 6-8 or 7-8.

[0105] In some embodiments of the present application, the pharmaceutical combination, wherein the average number of Linker-Payload conjugated to each antibody molecule (DAR) is 1-12, 1-11, 1-10, 1-9, 1-8, 2-8, 2-6, 2-4, 3-8, 4-8, 5-8, 6-8, 7-8, 1, 2, 3, 4, 5, 6, 7 or 8.

[0106] The mass ratio or molar ratio of the first active ingredient to the second active ingredient is (0.1-30): 1, preferably (0.3-20): 1, (0.4-15): 1, (0.5-15): 1, (0.5-10): 1, (0.5-5): 1, (0.8-5): 1, (0.8-3): 1, (1-3): 1, (1.2-3): 1, (1.5-2.5): 1, (1.8-2.2): 1, 0.1: 1, 0.2: 1, 0.3: 1, 0.4: 1, 0.5: 1, 0.6: 1, 0.7: 1, 0.8: 1, 0.9: 1, 1: 1, 1.1: 1, 1.2: 1, 1.3: 1, 1.4: 1, 1.5: 1, 1.6: 1, 1.7: 1, 1.8: 1, 1.9: 1, 2: 1, 2.1: 1, 2.2: 1, 2.3: 1, 2.4: 1, or 2.5: 1.

[0107] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0108] The first active ingredient is an antibody or an antigen-binding fragment thereof,

[0109] The second active ingredient is an antibody drug conjugate;

[0110] The antibody drug conjugate comprises the antibody or the antigen-binding fragment thereof, a linker, and a payload, wherein the linker-payload is a compound of Formula 0 or a pharmaceutically acceptable salt or ester thereof;

[0111] The compound of Formula 0 comprises a compound of Formula III, n alanines, and a compound of Formula IV connected in sequence,

[0112]

[0113] wherein,

[0114] n is 2, 3, 4, 5, or 6;

[0115] The compound of Formula III, the n alanines, and the compound of Formula IV are independently connected to each other two by two, directly or through a chemical group.

[0116] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0117] The first active ingredient is an anti-HER3 antibody or an antigen-binding fragment thereof,

[0118] The second active ingredient is an antibody drug conjugate;

[0119] The antibody drug conjugate comprises the anti-HER3 antibody or the antigen-binding fragment thereof, a linker, and a payload, wherein:

[0120] the linker is a hydrazone bond, a disulfide bond, a thioether bond or a peptide bond, or is a chemical group comprising a hydrazone bond, a disulfide bond, a thioether bond or a peptide bond;

[0121] Preferably, the linker is one or more selected from 6-maleimidocaproyl (MC), maleimidopropionyl (MP), valine-citrulline (Val-Cit), alanine-phenylalanine (Ala-Phe), alanine-alanine-alanine (Ala-Ala-Ala), Mc-Ala-Ala-Ala, p-aminobenzyloxycarbonyl (PAB), 6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (MC-Val-Cit-PAB), Mal-PEGn-Val-Cit-PAB (n is a positive integer selected from 1-20), Phe-Lys(Fmoc)-PAB, Aloc-D-Ala-Phe-Lys(Aloc)-PAB-PNP, Boc-Phe-(Alloc)Lys-PAB-PNP and 3-(pyridine-2-yl disulfide) propionic acid perfluorophenyl ester;

[0122] the payload is one or more selected from a tubulin inhibitor, a DNA damaging agent, a topoisomerase inhibitor, an ALK inhibitor and a PARP inhibitor;

[0123] Preferably, the tubulin inhibitor is one or more selected from dolastatins, auristatin cytotoxic molecules and maytansine cytotoxic molecules; preferably, the auristatin cytotoxic molecule is selected from monomethyl auristatin E (MMAE) and derivatives thereof or monomethyl auristatin F (MMAF) and derivatives thereof; preferably, the maytansine cytotoxic molecule is selected from DM1 and derivatives thereof and DM4 and derivatives thereof;

[0124] Preferably, the DNA damaging agent is one or more selected from calicheamicin, duocarmycin and antrmycin (PBD) derivatives;

[0125] Preferably, the topoisomerase inhibitor is selected from camptothecins and camptothecin derivatives; preferably, the camptothecin and camptothecin derivative is 7-ethyl-10-hydroxy camptothecin (SN-38) or DXd;

[0126] Preferably, the PARP inhibitor is niraparib.

[0127] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0128] The first active ingredient is an anti-HER3 antibody or an antigen-binding fragment thereof,

[0129] The second active ingredient is an antibody drug conjugate;

[0130] The antibody drug conjugate comprises the anti-HER3 antibody or the antigen-binding fragment thereof, a linker, and a payload, wherein the linker-payload is a compound of Formula 0 or a pharmaceutically acceptable salt or ester thereof;

[0131] The compound of Formula 0 comprises a compound of Formula III, n alanines, and a compound of Formula IV connected in sequence,

[0132]

[0133] wherein,

[0134] n is 2, 3, 4, 5, or 6;

[0135] The compound of Formula III, the n alanines, and the compound of Formula IV are independently connected to each other two by two, directly or through a chemical group.

[0136] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0137] The first active ingredient is an anti-HER3 antibody or an antigen-binding fragment thereof,

[0138] The second active ingredient is an antibody drug conjugate;

[0139] The antibody drug conjugate comprises the anti-HER3 antibody or the antigen-binding fragment thereof, a linker, and a payload, wherein the linker-payload is a compound of Formula 0 or a pharmaceutically acceptable salt or ester thereof;

[0140] The compound of Formula 0 is shown in Formula I below,

[0141]

[0142] wherein,

[0143] A represents an alanine;

[0144] m is 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12;

[0145] n is 2, 3, 4, 5, or 6.

[0146] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0147] the first active ingredient is an anti-HER3 antibody or an antigen-binding fragment thereof,

[0148] the second active ingredient is an antibody drug conjugate;

[0149] wherein the antibody drug conjugate comprises the anti-HER3 antibody or the antigen-binding fragment thereof, a linker, and a payload, wherein the linker-payload is a compound of Formula 0 or a pharmaceutically acceptable salt or ester thereof:

[0150] wherein the compound of Formula 0 is shown in Formula II below,

[0151]

[0152] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0153] the first active ingredient is an anti-TROP2 antibody or an antigen-binding fragment thereof,

[0154] the second active ingredient is an antibody drug conjugate;

[0155] wherein the antibody drug conjugate comprises the anti-TROP2 antibody or the antigen-binding fragment thereof, a linker, and a payload, wherein:

[0156] the linker is a hydrazone bond, a disulfide bond, a thioether bond, or a peptide bond, or a chemical group comprising a hydrazone bond, a disulfide bond, a thioether bond, or a peptide bond;

[0157] Preferably, the linker is one or more selected from the group consisting of 6-maleimidocaproyl (MC), maleimidopropionyl (MP), valine-citrulline (Val-Cit), alanine-phenylalanine (Ala-Phe), alanine-alanine-alanine (Ala-Ala-Ala), Mc-Ala-Ala-Ala, p-aminobenzyloxy carbonyl (PAB), 6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxy carbonyl (MC-Val-Cit-PAB), Mal-PEGn-Val-Cit-PAB (n is a positive integer selected from 1-20), Phe-Lys(Fmoc)-PAB, Aloc-D-Ala-Phe-Lys(Aloc)-PAB-PNP, Boc-Phe-(Alloc)Lys-PAB-PNP, and 3-(pyridine-2-yl disulfide) propionic acid perfluorophenyl ester;

[0158] the payload is one or more selected from the group consisting of a tubulin inhibitor, a DNA damaging agent, a topoisomerase inhibitor, an ALK inhibitor, and a PARP inhibitor;

[0159] Preferably, the tubulin inhibitor is one or more selected from the group consisting of dolastatins, auristatin cytotoxic molecules, and maytansine cytotoxic molecules; preferably, the auristatin cytotoxic molecule is selected from the group consisting of monomethyl auristatin E (MMAE) and derivatives thereof, and monomethyl auristatin F (MMAF) and derivatives thereof; preferably, the maytansine cytotoxic molecule is selected from the group consisting of DM1 and derivatives thereof, and DM4 and derivatives thereof.

[0160] Preferably, the DNA damaging agent is one or more selected from the group consisting of calicheamicin, duocarmycin, and an anthramycin derivative (PBD).

[0161] Preferably, the topoisomerase inhibitor is selected from the group consisting of camptothecins and camptothecin derivatives; preferably, the camptothecin and camptothecin derivative is 7-ethyl-10-hydroxy camptothecin (SN-38) or DXd.

[0162] Preferably, the PARP inhibitor is Niraparib.

[0163] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0164] the first active ingredient is an antibody or an antigen-binding fragment thereof,

[0165] the second active ingredient is an antibody conjugate drug;

[0166] wherein the antibody conjugate drug comprises the antibody or the antigen-binding fragment thereof, a linker, and a payload, and the linker-payload is mc-vc-PAB-MMAE.

[0167] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0168] the first active ingredient is an anti-TROP2 antibody or an antigen-binding fragment thereof,

[0169] the second active ingredient is an antibody conjugate drug;

[0170] wherein the antibody drug conjugate comprises the anti-TROP2 antibody or antigen binding fragment thereof, a linker, and a payload, wherein the linker-payload is mc-vc-PAB-MMAE.

[0171] In some embodiments of the present application, the pharmaceutical combination comprises an anti-HER3 antibody and an antibody drug conjugate of the anti-HER3 antibody.

[0172] In some embodiments of the present application, the pharmaceutical combination consists of an anti-HER3 antibody and an antibody drug conjugate of the anti-HER3 antibody.

[0173] In some embodiments of the present application, the pharmaceutical combination consists of an anti-HER3 antibody and an antibody drug conjugate of the anti-HER3 antibody.

[0174] In some embodiments of the present application, the pharmaceutical combination, wherein the anti-HER3 antibody comprises a heavy chain variable region and a light chain variable region,

[0175] wherein,

[0176] the heavy chain variable region comprises HCDR1 as set forth in SEQ ID NO: 1, HCDR2 as set forth in SEQ ID NO: 2, and HCDR3 as set forth in SEQ ID NO: 3; and

[0177] the light chain variable region comprises LCDR1 as set forth in SEQ ID NO: 4, LCDR2 as set forth in SEQ ID NO: 5, and LCDR3 as set forth in SEQ ID NO: 6.

[0178] In some embodiments of the present application, the pharmaceutical combination, wherein the anti-HER3 antibody comprises a heavy chain variable region and a light chain variable region,

[0179] In some embodiments of the present application, the pharmaceutical combination, wherein the anti-HER3 antibody comprises a heavy chain variable region and a light chain variable region,

[0180] In some embodiments of the present application, the pharmaceutical combination, wherein the anti-HER3 antibody comprises a heavy chain variable region and a light chain variable region,

[0181] In some embodiments of the present application, the pharmaceutical combination comprises an anti-TROP2 antibody and an antibody-drug conjugate of the anti-TROP2 antibody.

[0182] In some embodiments of the present application, the pharmaceutical combination consists of an anti-TROP2 antibody and an antibody-drug conjugate of the anti-TROP2 antibody.

[0183] In some embodiments of the present application, the pharmaceutical combination consists of an anti-TROP2 antibody and an antibody-drug conjugate of the anti-TROP2 antibody.

[0184] In some embodiments of the present application, the pharmaceutical combination, wherein the anti-HER3 antibody comprises a heavy chain variable region and a light chain variable region,

[0185] wherein,

[0186] the heavy chain variable region comprises HCDR1 of SEQ ID NO: 21, HCDR2 of SEQ ID NO: 22, and HCDR3 of SEQ ID NO: 23; and

[0187] the light chain variable region comprises LCDR1 of SEQ ID NO: 24, LCDR2 of SEQ ID NO: 25, and LCDR3 of SEQ ID NO: 26.

[0188] In some embodiments of the present application, the pharmaceutical combination, wherein the anti-TROP2 antibody comprises a heavy chain variable region and a light chain variable region, the amino acid sequence of the heavy chain variable region is as set forth in SEQ ID NO: 17, and the amino acid sequence of the light chain variable region is as set forth in SEQ ID NO: 18.

[0189] In some embodiments of the present application, the pharmaceutical combination, wherein the anti-TROP2 antibody comprises a heavy chain constant region and a light chain constant region, the heavy chain constant region is a heavy chain constant region of human IgG1, IgG2, IgG3 or IgG4 or a variant thereof, preferably a heavy chain constant region of human IgG1; and the light chain constant region is Ig kappa chain C region.

[0190] In some embodiments of the present application, the pharmaceutical combination, wherein the anti-TROP2 antibody comprises a heavy chain constant region and a light chain constant region, the amino acid sequence of the heavy chain constant region is as set forth in SEQ ID NO: 11 or SEQ ID NO: 19; and the amino acid sequence of the light chain constant region is as set forth in SEQ ID NO: 13.

[0191] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0192] the first active ingredient is an anti-HER3 antibody or an antigen-binding fragment thereof,

[0193] the second active ingredient is an antibody drug conjugate;

[0194] wherein the antibody drug conjugate comprises the anti-HER3 antibody or the antigen-binding fragment thereof, a linker, and a payload, wherein the linker-payload is a compound of Formula 0 or a pharmaceutically acceptable salt or ester thereof;

[0195] the compound of Formula 0 comprises a compound of Formula III, n alanines, and a compound of Formula IV connected in sequence,

[0196]

[0197] wherein,

[0198] n is 2, 3, 4, 5, or 6;

[0199] the compound of Formula III, the n alanines, and the compound of Formula IV are independently connected to each other directly or through a chemical group;

[0200] wherein the anti-HER3 antibody comprises a heavy chain variable region and a light chain variable region, wherein,

[0201] the heavy chain variable region comprises HCDR1 set forth in SEQ ID NO: 1, HCDR2 set forth in SEQ ID NO: 2, and HCDR3 set forth in SEQ ID NO: 3;

[0202] the light chain variable region comprises LCDR1 set forth in SEQ ID NO: 4, LCDR2 set forth in SEQ ID NO: 5, and LCDR3 set forth in SEQ ID NO: 6.

[0203] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0204] the first active ingredient is an anti-HER3 antibody or an antigen-binding fragment thereof,

[0205] the second active ingredient is an antibody drug conjugate;

[0206] wherein the antibody drug conjugate comprises the anti-HER3 antibody or the antigen-binding fragment thereof, a linker, and a payload, wherein the linker-payload is a compound of Formula 0 or a pharmaceutically acceptable salt or ester thereof;

[0207] wherein the compound of Formula 0 is shown in Formula I below,

[0208]

[0209] wherein,

[0210] A represents alanine;

[0211] m is 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12;

[0212] n is 2, 3, 4, 5, or 6;

[0213] wherein the anti-HER3 antibody comprises a heavy chain variable region and a light chain variable region, wherein

[0214] the heavy chain variable region comprises HCDR1 set forth in SEQ ID NO: 1, HCDR2 set forth in SEQ ID NO: 2, and HCDR3 set forth in SEQ ID NO: 3;

[0215] the light chain variable region comprises LCDR1 set forth in SEQ ID NO: 4, LCDR2 set forth in SEQ ID NO: 5, and LCDR3 set forth in SEQ ID NO: 6.

[0216] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0217] the first active ingredient is an anti-HER3 antibody or an antigen-binding fragment thereof,

[0218] the second active ingredient is an antibody drug conjugate;

[0219] wherein the antibody drug conjugate comprises the anti-HER3 antibody or the antigen-binding fragment thereof, a linker, and a payload, wherein the linker-payload is a compound of Formula 0 or a pharmaceutically acceptable salt or ester thereof;

[0220] wherein the compound of Formula 0 is shown below as Formula II,

[0221]

[0222] wherein the anti-HER3 antibody comprises a heavy chain variable region and a light chain variable region, wherein

[0223] the heavy chain variable region comprises HCDR1 set forth in SEQ ID NO: 1, HCDR2 set forth in SEQ ID NO: 2, and HCDR3 set forth in SEQ ID NO: 3;

[0224] the light chain variable region comprises LCDR1 set forth in SEQ ID NO: 4, LCDR2 set forth in SEQ ID NO: 5, and LCDR3 set forth in SEQ ID NO: 6.

[0225] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0226] the first active ingredient is an anti-HER3 antibody or an antigen-binding fragment thereof,

[0227] the second active ingredient is an antibody drug conjugate;

[0228] wherein the antibody drug conjugate comprises the anti-HER3 antibody or the antigen-binding fragment thereof, a linker, and a payload, wherein the linker-payload is a compound of Formula 0 or a pharmaceutically acceptable salt or ester thereof;

[0229] wherein the compound of Formula 0 is shown below as Formula II,

[0230]

[0231] wherein the anti-HER3 antibody comprises a heavy chain variable region and a light chain variable region, wherein:

[0232] the heavy chain variable region comprises HCDR1 of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, and HCDR3 of SEQ ID NO: 3;

[0233] the light chain variable region comprises LCDR1 of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO: 6;

[0234] wherein the average number of linker-payload conjugated to each anti-HER3 antibody molecule is 1-8;

[0235] Preferably, the mass ratio or molar ratio of the first active ingredient to the second active ingredient is (1:2) to (2:1).

[0236] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0237] the first active ingredient is an anti-HER3 antibody or an antigen-binding fragment thereof,

[0238] the second active ingredient is an antibody drug conjugate;

[0239] wherein the antibody drug conjugate comprises the anti-HER3 antibody or the antigen-binding fragment thereof, a linker, and a payload, wherein the linker-payload is a compound of Formula 0 or a pharmaceutically acceptable salt or ester thereof;

[0240] wherein the compound of Formula 0 is shown below as Formula II,

[0241]

[0242] wherein the anti-HER3 antibody comprises a heavy chain variable region and a light chain variable region, wherein,

[0243] the amino acid sequence of the heavy chain variable region of the anti-HER3 antibody is set forth in SEQ ID NO: 7, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 9;

[0244] wherein the average number of linker-payload conjugated to each anti-HER3 antibody molecule is 1-8;

[0245] Preferably, the mass ratio or molar ratio of the first active ingredient to the second active ingredient is (1:2) to (2:1).

[0246] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0247] the first active ingredient is an anti-HER3 antibody or an antigen-binding fragment thereof,

[0248] the second active ingredient is an antibody-drug conjugate;

[0249] wherein the antibody-drug conjugate comprises the anti-HER3 antibody or the antigen-binding fragment thereof, a linker, and a payload, wherein the linker-payload is a compound of Formula 0 or a pharmaceutically acceptable salt or ester thereof;

[0250] wherein the compound of Formula 0 is shown in Formula II below,

[0251]

[0252] wherein the anti-HER3 antibody comprises a heavy chain variable region and a light chain variable region, wherein,

[0253] the amino acid sequence of the heavy chain variable region of the anti-HER3 antibody is set forth in SEQ ID NO: 7 and the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO: 11, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 9 and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO: 13;

[0254] wherein the average number of linker-payload conjugated to each anti-HER3 antibody molecule is 1-8;

[0255] Preferably, the mass ratio or molar ratio of the first active ingredient to the second active ingredient is (1:2) to (2:1).

[0256] In some embodiments of the application, the pharmaceutical combination, wherein the average number of linkers-payloads conjugated to each anti-HER3 antibody molecule (DAR) is 1-8, 2-8, 2-6, 2-4, 3-8, 4-8, 5-8, 6-8, 7-8, 1, 2, 3, 4, 5, 6, 7, or 8.

[0257] In some embodiments of the application, the pharmaceutical combination, wherein the average number of linkers-payloads conjugated to each anti-HER3 antibody molecule (DAR) is 1-8, 2-8, 2-6, 2-4, 3-8, 4-8, 5-8, 6-8, 7-8, 1, 2, 3, 4, 5, 6, 7, or 8.

[0258] The mass ratio or molar ratio of the first active ingredient to the second active ingredient (mass ratio or molar ratio of the anti-HER3 antibody to the antibody-drug conjugate against HER3) is (0.1-30): 1 or (1:2) to (2:1), preferably (0.3-20): 1, (0.4-15): 1, (0.5-15): 1, (0.5-10): 1, (0.5-5): 1, (0.8-5): 1, (0.8-3): 1, (1-4): 1, (1-3): 1, (1.2-3): 1, (1.5-2.5): 1, (1.8-2.2): 1, 0.1: 1, 0.2: 1, 0.3: 1, 0.4: 1, 0.5: 1, 0.6: 1, 0.7: 1, 0.8: 1, 0.9: 1, 1: 1, 1.1: 1, 1.2: 1, 1.3: 1, 1.4: 1, 1.5: 1, 1.6: 1, 1.7: 1, 1.8: 1, 1.9: 1, 2: 1, 2.1: 1, 2.2: 1, 2.3: 1, 2.4: 1, or 2.5: 1.

[0259] In some embodiments of the application, the pharmaceutical combination, wherein the average number of linkers-payloads conjugated to each anti-HER3 antibody molecule (DAR) is 1-8, 2-8, 2-6, 2-4, 3-8, 4-8, 5-8, 6-8, 7-8, 1, 2, 3, 4, 5, 6, 7, or 8.

[0260] The mass ratio or molar ratio of the first active ingredient to the second active ingredient (mass ratio or molar ratio of the anti-HER3 antibody to the antibody-drug conjugate against HER3) is 1: (0.1-30), preferably 1: (0.3-20), 1: (0.4-15), 1: (0.5-15), 1: (0.5-10), 1: (0.5-5), 1: (0.8-5), 1: (0.8-3), 1: (1-4), 1: (1-3), 1: (1.2-3), 1: (1.5-2.5), 1: (1.8-2.2), 1: 0.1, 1: 0.2, 1: 0.3, 1: 0.4, 1: 0.5, 1: 0.6, 1: 0.7, 1: 0.8, 1: 0.9, 1: 1, 1: 1.1, 1: 1.2, 1: 1.3, 1: 1.4, 1: 1.5, 1: 1.6, 1: 1.7, 1: 1.8, 1: 1.9, 1: 2, 1: 2.1, 1: 2.2, 1: 2.3, 1: 2.4, or 1: 2.5.

[0261] In some embodiments of the application, the pharmaceutical combination comprises KA0013 and Patritumab.

[0262] In some embodiments of the application, the active ingredients of the pharmaceutical combination consist of KA0013 and Patritumab.

[0263] In some embodiments of the application, the pharmaceutical combination consists of KA0013 and Patritumab.

[0264] In some embodiments of the application, the pharmaceutical combination, comprises a first active ingredient and a second active ingredient, wherein:

[0265] the first active ingredient is an anti-TROP2 antibody or an antigen-binding fragment thereof,

[0266] the second active ingredient is an antibody drug conjugate;

[0267] wherein the antibody drug conjugate comprises the anti-TROP2 antibody or the antigen-binding fragment thereof, a linker, and a payload, wherein the linker is one or more selected from 6-maleimidocaproyl (MC), valine-citrulline (Val-Cit), and p-aminobenzyloxycarbonyl (PAB); and the payload is MMAE or a derivative thereof, or MMAF or a derivative thereof.

[0268] wherein the anti-TROP2 antibody comprises a heavy chain variable region and a light chain variable region, wherein:

[0269] the heavy chain variable region comprises HCDR1 as set forth in SEQ ID NO: 21, HCDR2 as set forth in SEQ ID NO: 22, and HCDR3 as set forth in SEQ ID NO: 23;

[0270] the light chain variable region comprises LCDR1 as set forth in SEQ ID NO: 24, LCDR2 as set forth in SEQ ID NO: 25, and LCDR3 as set forth in SEQ ID NO: 26.

[0271] In some embodiments of the application, the pharmaceutical combination, comprises a first active ingredient and a second active ingredient, wherein:

[0272] the first active ingredient is an anti-TROP2 antibody or an antigen-binding fragment thereof,

[0273] the second active ingredient is an antibody drug conjugate;

[0274] wherein the antibody drug conjugate comprises the anti-TROP2 antibody or the antigen-binding fragment thereof, a linker, and a payload, wherein the linker is one or more selected from 6-maleimidocaproyl (MC), valine-citrulline (Val-Cit), and p-aminobenzyloxycarbonyl (PAB); and the payload is MMAE or a derivative thereof, or MMAF or a derivative thereof.

[0275] wherein the anti-TROP2 antibody comprises a heavy chain variable region and a light chain variable region, wherein:

[0276] the heavy chain variable region comprises HCDR1 set forth in SEQ ID NO: 21, HCDR2 set forth in SEQ ID NO: 22, and HCDR3 set forth in SEQ ID NO: 23;

[0277] the light chain variable region comprises LCDR1 set forth in SEQ ID NO: 24, LCDR2 set forth in SEQ ID NO: 25, and LCDR3 set forth in SEQ ID NO: 26.

[0278] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0279] the first active ingredient is an anti-TROP2 antibody or an antigen-binding fragment thereof,

[0280] the second active ingredient is an antibody drug conjugate;

[0281] wherein the antibody drug conjugate comprises the anti-TROP2 antibody or the antigen-binding fragment thereof, a linker, and a payload, wherein the linker-payload is mc-vc-PAB-MMAE.

[0282] wherein the anti-TROP2 antibody comprises a heavy chain variable region and a light chain variable region, wherein:

[0283] the heavy chain variable region comprises HCDR1 set forth in SEQ ID NO: 21, HCDR2 set forth in SEQ ID NO: 22, and HCDR3 set forth in SEQ ID NO: 23;

[0284] the light chain variable region comprises LCDR1 set forth in SEQ ID NO: 24, LCDR2 set forth in SEQ ID NO: 25, and LCDR3 set forth in SEQ ID NO: 26.

[0285] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0286] the first active ingredient is an anti-TROP2 antibody or an antigen-binding fragment thereof,

[0287] the second active ingredient is an antibody drug conjugate;

[0288] The antibody drug conjugate comprises the anti-TROP2 antibody or antigen binding fragment thereof, a linker, and a payload, wherein the payload is MMAE or a derivative thereof, or MMAF or a derivative thereof.

[0289] The anti-TROP2 antibody comprises a heavy chain variable region and a light chain variable region, wherein,

[0290] The heavy chain variable region comprises HCDR1 as set forth in SEQ ID NO: 21, HCDR2 as set forth in SEQ ID NO: 22, and HCDR3 as set forth in SEQ ID NO: 23.

[0291] The light chain variable region comprises LCDR1 as set forth in SEQ ID NO: 24, LCDR2 as set forth in SEQ ID NO: 25, and LCDR3 as set forth in SEQ ID NO: 26.

[0292] The average number of linker-payload conjugated to each anti-TROP2 antibody molecule is 1-8.

[0293] Preferably, the mass ratio or molar ratio of the first active ingredient to the second active ingredient is (1:2) to (2:1).

[0294] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0295] The first active ingredient is an anti-TROP2 antibody or antigen binding fragment thereof,

[0296] The second active ingredient is an antibody drug conjugate.

[0297] The antibody drug conjugate comprises the anti-TROP2 antibody or antigen binding fragment thereof, a linker, and a payload, wherein the linker is one or more selected from 6-maleimidocaproyl (MC), valine-citrulline (Val-Cit), and p-aminobenzyloxy carbonyl (PAB); and the payload is MMAE or a derivative thereof, or MMAF or a derivative thereof.

[0298] The anti-TROP2 antibody comprises a heavy chain variable region and a light chain variable region, wherein,

[0299] The heavy chain variable region comprises HCDR1 as set forth in SEQ ID NO: 21, HCDR2 as set forth in SEQ ID NO: 22, and HCDR3 as set forth in SEQ ID NO: 23.

[0300] the heavy chain variable region comprises HCDR1 set forth in SEQ ID NO: 21, HCDR2 set forth in SEQ ID NO: 22, and HCDR3 set forth in SEQ ID NO: 23;

[0301] wherein the average number of linker-payload conjugated to each anti-TROP2 antibody molecule is 1-8;

[0302] Preferably, the mass ratio or molar ratio of the first active ingredient to the second active ingredient is (1:2) to (2:1).

[0303] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0304] the first active ingredient is an anti-TROP2 antibody or an antigen-binding fragment thereof,

[0305] the second active ingredient is an antibody-drug conjugate;

[0306] wherein the antibody-drug conjugate comprises the anti-TROP2 antibody or the antigen-binding fragment thereof, a linker, and a payload, wherein the linker-payload is mc-vc-PAB-MMAE;

[0307] wherein the anti-TROP2 antibody comprises a heavy chain variable region and a light chain variable region, wherein,

[0308] the heavy chain variable region comprises HCDR1 set forth in SEQ ID NO: 21, HCDR2 set forth in SEQ ID NO: 22, and HCDR3 set forth in SEQ ID NO: 23;

[0309] the light chain variable region comprises LCDR1 set forth in SEQ ID NO: 24, LCDR2 set forth in SEQ ID NO: 25, and LCDR3 set forth in SEQ ID NO: 26;

[0310] wherein the average number of linker-payload conjugated to each anti-TROP2 antibody molecule is 1-8;

[0311] Preferably, the mass ratio or molar ratio of the first active ingredient to the second active ingredient is (1:2) to (2:1).

[0312] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0313] the first active ingredient is an anti-TROP2 antibody or an antigen-binding fragment thereof,

[0314] the second active ingredient is an antibody-drug conjugate;

[0315] wherein the antibody drug conjugate comprises the anti-TROP2 antibody or antigen binding fragment thereof, a linker, and a payload, wherein the payload is MMAE or a derivative thereof, or MMAF or a derivative thereof;

[0316] wherein the anti-TROP2 antibody comprises a heavy chain variable region and a light chain variable region, wherein,

[0317] the amino acid sequence of the heavy chain variable region of the anti-TROP2 antibody is set forth in SEQ ID NO: 17, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 18;

[0318] wherein the average number of linker-payload conjugated to each anti-TROP2 antibody molecule is 1-8;

[0319] Preferably, the mass ratio or molar ratio of the first active ingredient to the second active ingredient is (1:2) to (2:1).

[0320] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0321] the first active ingredient is an anti-TROP2 antibody or antigen binding fragment thereof,

[0322] the second active ingredient is an antibody drug conjugate;

[0323] wherein the antibody drug conjugate comprises the anti-TROP2 antibody or antigen binding fragment thereof, a linker, and a payload, wherein the linker is one or more selected from the group consisting of 6-maleimidocaproyl (MC), valine-citrulline (Val-Cit), and p-aminobenzyloxycarbonyl (PAB); and the payload is MMAE or a derivative thereof, or MMAF or a derivative thereof;

[0324] wherein the anti-TROP2 antibody comprises a heavy chain variable region and a light chain variable region, wherein,

[0325] the amino acid sequence of the heavy chain variable region of the anti-TROP2 antibody is set forth in SEQ ID NO: 17, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 18;

[0326] wherein the average number of linker-payload conjugated to each anti-TROP2 antibody molecule is 1-8;

[0327] Preferably, the mass ratio or molar ratio of the first active ingredient to the second active ingredient is (1:2) to (2:1).

[0328] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0329] the first active ingredient is an anti-TROP2 antibody or an antigen-binding fragment thereof,

[0330] the second active ingredient is an antibody drug conjugate;

[0331] wherein the antibody drug conjugate comprises the anti-TROP2 antibody or the antigen-binding fragment thereof, a linker, and a payload, and the linker-payload is mc-vc-PAB-MMAE;

[0332] wherein the anti-TROP2 antibody comprises a heavy chain variable region and a light chain variable region, wherein:

[0333] the amino acid sequence of the heavy chain variable region of the anti-TROP2 antibody is set forth in SEQ ID NO: 17, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 18;

[0334] wherein the average number of linker-payload conjugated to each anti-TROP2 antibody molecule is 1-8;

[0335] Preferably, the mass ratio or molar ratio of the first active ingredient to the second active ingredient is (1:2) to (2:1).

[0336] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0337] the first active ingredient is an anti-TROP2 antibody or an antigen-binding fragment thereof,

[0338] the second active ingredient is an antibody drug conjugate;

[0339] wherein the antibody drug conjugate comprises the anti-TROP2 antibody or the antigen-binding fragment thereof, a linker, and a payload, and the payload is MMAE or a derivative thereof, or MMAF or a derivative thereof;

[0340] wherein the anti-TROP2 antibody comprises a heavy chain variable region and a light chain variable region, wherein:

[0341] the amino acid sequence of the heavy chain variable region of the anti-TROP2 antibody is set forth in SEQ ID NO: 17 and the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO: 19, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 18 and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO: 13;

[0342] wherein the average number of linker-payload conjugated to each anti-TROP2 antibody molecule is 1-8;

[0343] Preferably, the mass ratio or molar ratio of the first active ingredient to the second active ingredient is (1:2) to (2:1).

[0344] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0345] The first active ingredient is an anti-TROP2 antibody or an antigen-binding fragment thereof,

[0346] The second active ingredient is an antibody-drug conjugate;

[0347] wherein the antibody-drug conjugate comprises the anti-TROP2 antibody or the antigen-binding fragment thereof, a linker, and a payload, wherein the linker is one or more selected from 6-maleimidocaproyl (MC), valine-citrulline (Val-Cit), and p-aminobenzyloxycarbonyl (PAB); and the payload is MMAE or a derivative thereof, or MMAF or a derivative thereof.

[0348] wherein the anti-TROP2 antibody comprises a heavy chain variable region and a light chain variable region, wherein,

[0349] the amino acid sequence of the heavy chain variable region of the anti-TROP2 antibody is as set forth in SEQ ID NO: 17 and the amino acid sequence of the heavy chain constant region is as set forth in SEQ ID NO: 19, and the amino acid sequence of the light chain variable region is as set forth in SEQ ID NO: 18 and the amino acid sequence of the light chain constant region is as set forth in SEQ ID NO: 13;

[0350] wherein the average number of linker-payload conjugated to each anti-TROP2 antibody molecule is 1-8;

[0351] Preferably, the mass ratio or molar ratio of the first active ingredient to the second active ingredient is (1:2) to (2:1).

[0352] In some embodiments of the present application, the pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein:

[0353] The first active ingredient is an anti-TROP2 antibody or an antigen-binding fragment thereof,

[0354] The second active ingredient is an antibody-drug conjugate;

[0355] wherein the antibody-drug conjugate comprises the anti-TROP2 antibody or the antigen-binding fragment thereof, a linker, and a payload, wherein the linker-payload is mc-vc-PAB-MMAE.

[0356] wherein the anti-TROP2 antibody comprises a heavy chain variable region and a light chain variable region, wherein,

[0357] the amino acid sequence of the heavy chain variable region of the anti-TROP2 antibody is set forth in SEQ ID NO: 17 and the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO: 19, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 18 and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO: 13;

[0358] wherein the average number of Linker-Payload conjugated to each anti-TROP2 antibody molecule is 1-8;

[0359] Preferably, the mass ratio or molar ratio of the first active ingredient to the second active ingredient is (1:2) to (2:1).

[0360] In some embodiments of the present application, the pharmaceutical combination, wherein the average number of Linker-Payload conjugated to each anti-TROP2 antibody molecule (DAR) is 1-8, 2-8, 2-6, 2-4, 3-8, 4-8, 5-8, 6-8, 7-8, 1, 2, 3, 4, 5, 6, 7 or 8.

[0361] In some embodiments of the present application, the pharmaceutical combination, wherein,

[0362] The mass ratio or molar ratio of the first active ingredient to the second active ingredient (mass ratio or molar ratio of anti-TROP2 antibody to anti-TROP2 antibody conjugate) is (0.1-30):1 or (1:2) to (2:1), preferably (0.3-20):1, (0.4-15):1, (0.5-15):1, (0.5-10):1, (0.5-5):1, (0.8-5):1, (0.8-3):1, (1-4):1, (1-3):1, (1.2-3):1, (1.5-2.5):1, (1.8-2.2):1, 0.1:1, 0.2:1, 0.3:1, 0.4:1, 0.5:1, 0.6:1, 0.7:1, 0.8:1, 0.9:1, 1:1, 1.1:1, 1.2:1, 1.3:1, 1.4:1, 1.5:1, 1.6:1, 1.7:1, 1.8:1, 1.9:1, 2:1, 2.1:1, 2.2:1, 2.3:1, 2.4:1 or 2.5:1.

[0363] In some embodiments of the present application, the pharmaceutical combination, wherein,

[0364] The mass ratio or molar ratio of the first active ingredient to the second active ingredient (mass ratio or molar ratio of the anti-TROP2 antibody to the antibody-drug conjugate against TROP2) is 1 : (0.1-30), preferably 1 : (0.3-20), 1 : (0.4-15), 1 : (0.5-15), 1 : (0.5-10), 1 : (0.5-5), 1 : (0.8-5), 1 : (0.8-3), 1 : (1-4), 1 : (1-3), 1 : (1.2-3), 1 : (1.5-2.5), 1 : (1.8-2.2), 1 :0.1, 1 :0.2, 1 :0.3, 1 :0.4, 1 :0.5, 1 :0.6, 1 :0.7, 1 :0.8, 1 :0.9, 1 :1, 1 :1.1, 1 :1.2, 1 :1.3, 1 :1.4, 1 :1.5, 1 :1.6, 1 :1.7, 1 :1.8, 1 :1.9, 1 :2, 1 :2.1, 1 :2.2, 1 :2.3, 1 :2.4, or 1 :2.5.

[0365] In some embodiments of the application, the pharmaceutical combination comprises hRS7(G1TM) and hRS7(G1TM)-mc-vc-PAB-MMAE.

[0366] In some embodiments of the application, the active ingredients of the pharmaceutical combination consist of hRS7(G1TM) and hRS7(G1TM)-mc-vc-PAB-MMAE.

[0367] In some embodiments of the application, the pharmaceutical combination consists of hRS7(G1TM) and hRS7(G1TM)-mc-vc-PAB-MMAE.

[0368] The pharmaceutical combination of the application can be used for the treatment or prevention of a tumor or an autoimmune disease;

[0369] Preferably, the tumor is a HER3-positive or TROP2-positive tumor.

[0370] Preferably, the tumor is selected from one or more of lung cancer, colon cancer, rectal cancer, breast cancer, clear cell sarcoma, skin cancer, renal cancer, urothelial cancer, prostate cancer, glioblastoma multiforme, ovarian cancer, pancreatic cancer, melanoma, liver cancer, bladder cancer, gastric cancer, gastrointestinal stromal tumor, cervical cancer, head and neck cancer, esophageal cancer, epidermoid cancer, peritoneal cancer, adult glioblastoma multiforme, uterine cancer, salivary gland cancer, vulvar cancer, thyroid cancer, anal cancer, and penile cancer.

[0371] Preferably, the lung cancer is non-small cell lung cancer (NSCLC), e.g. lung adenocarcinoma.

[0372] Preferably, the colon cancer is metastatic colon cancer.

[0373] Preferably, the autoimmune disease is one or more selected from systemic lupus erythematosus, rheumatoid arthritis, Sjogren's syndrome, ankylosing spondylitis, multiple sclerosis, autoimmune thyroid disease, and inflammatory bowel disease.

[0374] In some embodiments of the present application, the pharmaceutical combination, wherein the first active ingredient and the second active ingredient are mixed together, the pharmaceutical combination is a pharmaceutical composition; or

[0375] the first active ingredient and the second active ingredient are in separate packages from each other, the pharmaceutical combination is a combination product;

[0376] Preferably, the pharmaceutical combination further comprises one or more pharmaceutically acceptable excipients.

[0377] In some embodiments of the present application, the pharmaceutical combination, wherein the dosage is 0.1 mg / kg-15 mg / kg or 1 mg / kg-15 mg / kg, calculated according to the mass (mg) of the second active ingredient (antibody conjugate drug) in the pharmaceutical combination and the body weight (kg) of the subject;

[0378] Preferably, the administration dosage is 0.1 mg / kg, 0.2 mg / kg, 0.3 mg / kg, 0.4 mg / kg, 0.5 mg / kg, 0.6 mg / kg, 0.7 mg / kg, 0.8 mg / kg, 0.9 mg / kg, 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 mg / kg, 11.2 mg / kg, 11.4 mg / kg, 11.6 mg / kg, 11.8 mg / kg, 12 mg / kg, 12.2 mg / kg, 12.4 mg / kg, 12.6 mg / kg, 12.8 mg / kg, 13 mg / kg, 13.2 mg / kg, 13.4 mg / kg, 13.6 mg / kg, 13.8 mg / kg, 14 mg / kg, 14.2 mg / kg, 14.4 mg / kg, 14.6 mg / kg, 14.8 mg / kg, or 15 mg / kg.

[0379] Preferably, the administration frequency of the drug combination (including the first active ingredient and / or the second active ingredient) is once every 1 week, once every 2 weeks, once every 3 weeks, once every 4 weeks, once every 5 weeks, once every 6 weeks, once every 1 month, once every 2 months, once every 3 months, once every 4 months, once every 5 months, or once every 6 months.

[0380] Preferably, the administration mode is intratumoral injection, intravenous drip, intravenous injection, subcutaneous injection, intraperitoneal injection, or oral administration.

[0381] In some embodiments of the present application, the pharmaceutical combination, wherein the administration dosage of the second active ingredient (antibody conjugate drug) is half of the administration dosage of the first active ingredient (antibody or antigen binding fragment thereof) in the pharmaceutical combination, for example, 0.05 mg / kg-7.5 mg / kg or 0.5 mg / kg-7.5 mg / kg, calculated by the mass (mg) of the first active ingredient (antibody or antigen binding fragment thereof) and the weight (kg) of the subject; the preferred administration dosage of the second active ingredient (antibody conjugate drug) can also be calculated by half of the preferred administration dosage of the second active ingredient (antibody conjugate drug), for example: 0.05 mg / kg, 0.1 mg / kg, 0.15 mg / kg, …, 7.2 mg / kg, 7.3 mg / kg, 7.4 mg / kg, or 7.5 mg / kg.

[0382] Another aspect of the present application relates to a packaged product comprising an effective amount of the pharmaceutical combination of any one of the present application, and a packaging container;

[0383] Preferably, the packaging container is a syringe or an injection pen.

[0384] Preferably, the packaged product further comprises a product instruction.

[0385] Another aspect of the present application relates to a pharmaceutical composition comprising the pharmaceutical combination of any one of the present application, and one or more pharmaceutically acceptable excipients.

[0386] In some embodiments of the present application, the pharmaceutical composition, wherein the active ingredients thereof consist of the first active ingredient and the second active ingredient.

[0387] In some embodiments of the present application, the pharmaceutical composition, wherein the active ingredient thereof is the pharmaceutical combination of the present application.

[0388] In some embodiments of the present application, the pharmaceutical composition, wherein the active ingredient thereof consists of the pharmaceutical combination of the present application and one or more pharmaceutically acceptable excipients.

[0389] The pharmaceutical composition of the present application can be used for treating or preventing a tumor or an autoimmune disease.

[0390] Preferably, the tumor is a HER3-positive or TROP2-positive tumor.

[0391] Preferably, the tumor is selected from one or more of lung cancer, colon cancer, rectal cancer, breast cancer, clear cell sarcoma, skin cancer, renal cancer, urothelial cancer, prostate cancer, glioblastoma multiforme, ovarian cancer, pancreatic cancer, melanoma, liver cancer, bladder cancer, gastric cancer, gastrointestinal stromal tumor, cervical cancer, head and neck cancer, esophageal cancer, epidermoid cancer, peritoneal cancer, adult glioblastoma multiforme, uterine cancer, salivary gland cancer, vulvar cancer, thyroid cancer, anal cancer, and penile cancer.

[0392] Preferably, the lung cancer is non-small cell lung cancer (NSCLC) such as lung adenocarcinoma;

[0393] Preferably, the colon cancer is metastatic colon cancer.

[0394] Preferably, the autoimmune disease is one or more selected from systemic lupus erythematosus, rheumatoid arthritis, Sjogren's syndrome, ankylosing spondylitis, multiple sclerosis, autoimmune thyroid disease, and inflammatory bowel disease.

[0395] In some embodiments of the present application, the pharmaceutical composition, wherein the administration dosage is 0.1 mg / kg to 15 mg / kg or 1 mg / kg to 15 mg / kg, calculated by the mass (mg) of the second active ingredient (antibody conjugate drug) in the pharmaceutical composition and the body weight (kg) of the subject;

[0396] Preferably, the administration dosage is 0.1 mg / kg, 0.2 mg / kg, 0.3 mg / kg, 0.4 mg / kg, 0.5 mg / kg, 0.6 mg / kg, 0.7 mg / kg, 0.8 mg / kg, 0.9 mg / kg, 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 mg / kg, 11.2 mg / kg, 11.4 mg / kg, 11.6 mg / kg, 11.8 mg / kg, 12 mg / kg, 12.2 mg / kg, 12.4 mg / kg, 12.6 mg / kg, 12.8 mg / kg, 13 mg / kg, 13.2 mg / kg, 13.4 mg / kg, 13.6 mg / kg, 13.8 mg / kg, 14 mg / kg, 14.2 mg / kg, 14.4 mg / kg, 14.6 mg / kg, 14.8 mg / kg, or 15 mg / kg.

[0397] Preferably, the administration frequency of the pharmaceutical composition (including the first active ingredient and / or the second active ingredient) is once every 1 week, once every 2 weeks, once every 3 weeks, once every 4 weeks, once every 5 weeks, once every 6 weeks, once every 1 month, once every 2 months, once every 3 months, once every 4 months, once every 5 months, or once every 6 months.

[0398] Preferably, the administration mode is intratumoral injection, intravenous drip, intravenous injection, subcutaneous injection, intraperitoneal injection, or oral administration.

[0399] In some embodiments of the present application, the pharmaceutical composition, wherein the administration dosage of the second active ingredient (antibody conjugate drug) is half of the above-mentioned preferred administration dosage, for example, 0.05 mg / kg-7.5 mg / kg or 0.5 mg / kg-7.5 mg / kg; preferably, the administration dosage of the second active ingredient (antibody conjugate drug) can also be calculated as half of the above-mentioned preferred administration dosage, for example: 0.05 mg / kg, 0.1 mg / kg, 0.15 mg / kg, …, 7.2 mg / kg, 7.3 mg / kg, 7.4 mg / kg, or 7.5 mg / kg.

[0400] Still another aspect of the present application relates to the use of the pharmaceutical combination of any one of the present application or the pharmaceutical composition of the present application in the preparation of a medicament for treating or preventing a tumor or an autoimmune disease;

[0401] Preferably, the tumor is a HER3-positive or TROP2-positive tumor.

[0402] Preferably, the tumor is selected from one or more of lung cancer, colon cancer, rectal cancer, breast cancer, clear cell sarcoma, skin cancer, renal cancer, urothelial cancer, prostate cancer, glioblastoma multiforme, ovarian cancer, pancreatic cancer, melanoma, liver cancer, bladder cancer, gastric cancer, gastrointestinal stromal tumor, cervical cancer, head and neck cancer, esophageal cancer, epidermoid cancer, peritoneal cancer, adult glioblastoma multiforme, uterine cancer, salivary gland cancer, vulvar cancer, thyroid cancer, anal cancer, and penile cancer.

[0403] Preferably, the lung cancer is non-small cell lung cancer, for example, lung adenocarcinoma.

[0404] Preferably, the colon cancer is metastatic colon cancer.

[0405] Preferably, the autoimmune disease is selected from one or more of systemic lupus erythematosus, rheumatoid arthritis, Sjogren's syndrome, ankylosing spondylitis, multiple sclerosis, autoimmune thyroid disease, and inflammatory bowel disease.

[0406] In some embodiments of the present application, the use, wherein the administration dosage of the second active ingredient (antibody conjugate drug) is 0.1 mg / kg-15 mg / kg or 1 mg / kg-15 mg / kg, calculated by the mass (mg) of the second active ingredient (antibody conjugate drug) in the pharmaceutical combination and the body weight (kg) of the subject;

[0407] Preferably, the dosage administered is 0.1 mg / kg, 0.2 mg / kg, 0.3 mg / kg, 0.4 mg / kg, 0.5 mg / kg, 0.6 mg / kg, 0.7 mg / kg, 0.8 mg / kg, 0.9 mg / kg, 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 mg / kg, 11.2 mg / kg, 11.4 mg / kg, 11.6 mg / kg, 11.8 mg / kg, 12 mg / kg, 12.2 mg / kg, 12.4 mg / kg, 12.6 mg / kg, 12.8 mg / kg, 13 mg / kg, 13.2 mg / kg, 13.4 mg / kg, 13.6 mg / kg, 13.8 mg / kg, 14 mg / kg, 14.2 mg / kg, 14.4 mg / kg, 14.6 mg / kg, 14.8 mg / kg, or 15 mg / kg.

[0408] Preferably, the frequency of administration of the pharmaceutical combination (including the first active ingredient and / or the second active ingredient) is once every 1 week, once every 2 weeks, once every 3 weeks, once every 4 weeks, once every 5 weeks, once every 6 weeks, once every 1 month, once every 2 months, once every 3 months, once every 4 months, once every 5 months, or once every 6 months.

[0409] Preferably, the mode of administration is intratumoral injection, intravenous infusion, intravenous injection, subcutaneous injection, intraperitoneal injection, or oral administration.

[0410] In some embodiments of the present invention, the dosage is calculated based on the mass (mg) of the first active ingredient (antibody or its antigen-binding fragment) in the drug combination and the subject's weight (kg), and is half of the second active ingredient (antibody-drug conjugate), for example, 0.05 mg / kg-7.5 mg / kg or 0.5 mg / kg-7.5 mg / kg; the preferred dosage may also be calculated with reference to half of the preferred dosage of the second active ingredient (antibody-drug conjugate), for example: 0.05 mg / kg, 0.1 mg / kg, 0.15 mg / kg, ..., 7.2 mg / kg, 7.3 mg / kg, 7.4 mg / kg, or 7.5 mg / kg.

[0411] Another aspect of the invention relates to a method for treating or preventing tumors or autoimmune diseases, comprising the step of administering to a subject in need an effective amount of any of the pharmaceutical combinations or pharmaceutical compositions of the present invention.

[0412] Preferably, the tumor is a HER3-positive or TROP2-positive tumor;

[0413] Preferably, the tumor is selected from one or more of the following: lung cancer, colon cancer, rectal cancer, breast cancer, clear cell sarcoma, skin cancer, kidney cancer, urothelial carcinoma, prostate cancer, glioblastoma multiforme, ovarian cancer, pancreatic cancer, melanoma, liver cancer, bladder cancer, gastric cancer, gastrointestinal stromal tumor, cervical cancer, head and neck cancer, esophageal cancer, epidermoid carcinoma, peritoneal cancer, adult glioblastoma multiforme, uterine cancer, salivary gland cancer, vulvar cancer, thyroid cancer, anal cancer, and penile cancer.

[0414] Preferably, the lung cancer is non-small cell lung cancer (NSCLC), such as lung adenocarcinoma;

[0415] Preferably, the colon cancer is metastatic colon cancer;

[0416] Preferably, the autoimmune disease is selected from one or more of systemic lupus erythematosus, rheumatoid arthritis, Sjögren's syndrome, ankylosing spondylitis, multiple sclerosis, autoimmune thyroid disease, and inflammatory bowel disease.

[0417] In some embodiments of the present invention, the method for treating or preventing tumors is wherein the dosage is calculated based on the mass (mg) of the second active ingredient (antibody-drug conjugate) in the drug combination and the subject's weight (kg), and is 0.1 mg / kg-15 mg / kg or 1 mg / kg-15 mg / kg.

[0418] Preferably, the administration dose is 0.1 mg / kg, 0.2 mg / kg, 0.3 mg / kg, 0.4 mg / kg, 0.5 mg / kg, 0.6 mg / kg, 0.7 mg / kg, 0.8 mg / kg, 0.9 mg / kg, 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 mg / kg, 11.2 mg / kg, 11.4 mg / kg, 11.6 mg / kg, 11.8 mg / kg, 12 mg / kg, 12.2 mg / kg, 12.4 mg / kg, 12.6 mg / kg, 12.8 mg / kg, 13 mg / kg, 13.2 mg / kg, 13.4 mg / kg, 13.6 mg / kg, 13.8 mg / kg, 14 mg / kg, 14.2 mg / kg, 14.4 mg / kg, 14.6 mg / kg, 14.8 mg / kg, or 15 mg / kg.

[0419] Preferably, the administration frequency of the drug combination (including the first active ingredient and / or the second active ingredient) is once every 1 week, once every 2 weeks, once every 3 weeks, once every 4 weeks, once every 5 weeks, once every 6 weeks, once every 1 month, once every 2 months, once every 3 months, once every 4 months, once every 5 months, or once every 6 months.

[0420] Preferably, the administration mode is intratumoral injection, intravenous drip, intravenous injection, subcutaneous injection, intraperitoneal injection, or oral administration.

[0421] In some embodiments of the present application, the method for treating or preventing tumor, wherein the administration dosage of the second active ingredient (antibody conjugated drug) is half of the administration dosage of the first active ingredient (antibody or antigen binding fragment thereof) in the pharmaceutical combination, for example, 0.05 mg / kg-7.5 mg / kg or 0.5 mg / kg-7.5 mg / kg, calculated by the mass (mg) of the first active ingredient (antibody or antigen binding fragment thereof) and the weight (kg) of the subject; preferably, the administration dosage of the second active ingredient (antibody conjugated drug) can also be calculated by half of the preferred administration dosage of the second active ingredient (antibody conjugated drug), for example: 0.05 mg / kg, 0.1 mg / kg, 0.15 mg / kg, …, 7.2 mg / kg, 7.3 mg / kg, 7.4 mg / kg, or 7.5 mg / kg.

[0422] In the present application, unless otherwise specified, the scientific and technical terms used herein have the meanings commonly understood by a person skilled in the art. And, the cell culture, molecular genetics, nucleic acid chemistry, immunology laboratory operation steps used herein are the conventional steps widely used in the corresponding field. At the same time, in order to better understand the present application, the definitions and explanations of related terms are provided below.

[0423] In the present application, the term "therapeutic window" refers to the difference between the minimum effective dose and the toxic dose of a drug. Generally speaking, the wider the therapeutic window, the better the safety of the drug, and the narrower the therapeutic window, the worse the safety of the drug.

[0424] As used herein, the term EC 50 refers to the concentration for 50% of maximal effect, which refers to the concentration that can cause 50% of the maximum effect.

[0425] As used herein, the term "antibody" refers to an immunoglobulin molecule that is generally composed of two pairs of polypeptide chains (each pair having one "light" (L) chain and one "heavy" (H) chain). Antibody light chains can be assigned to a class, kappa and lambda. Heavy chains can be assigned to a class, mu, delta, gamma, alpha, or epsilon, and define a specificity of an antibody as IgM, IgD, IgG, IgA, and IgE, respectively. Within the light and heavy chains, the variable and constant regions are joined by a "J" region of about 12 or more amino acids, and the heavy chain also contains a "D" region of about 3 or more amino acids. Each heavy chain is composed of a heavy chain variable region (VH) and a heavy chain constant region (CH). The heavy chain constant region is comprised of three domains, CH1, CH2 and CH3. Each light chain is composed of a light chain variable region (VL) and a light chain constant region (CL). The light chain constant region is comprised of one domain, CL. The constant regions of the antibodies can mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system. The VH and VL regions can also be subdivided into regions of hypervariability, termed complementarity determining regions (CDRs), interspersed with regions that are more conserved, termed framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The variable regions of each heavy / light chain pair (VH and VL) form the antibody binding site. Assignment of amino acids to each region or domain follows the definition of Bethesda M.d., Kabat Sequences of Proteins of Immunological Interest (National Institutes of Health, (1987 and 1991)), or Chothia & Lesk J. Mol. Biol. 1987; 196:901-917; Chothia et al. Nature 1989; 342:878-883, or the IMGT numbering system definition, see Ehrenmann F, Kaas Q, Lefranc M P. IMGT / 3Dstructure-DB and IMGT / DomainGapAlign: a database and a tool for immunoglobulins or antibodies, T cell receptors, MHC, IgSF and MhcSF [J]. Nucleic acids research, 2009; 38(suppl_1):D301-D307.

[0426] The variable regions of both light and heavy chains determine antigen binding; the variable regions of each chain contain three hypervariable regions of about 10-15 amino acids each, referred to as Complementarity Determining Regions (CDRs), wherein the CDRs of the heavy chain (H) include HCDR1, HCDR2, HCDR3, and the CDRs of the light chain (L) include LCDR1, LCDR2, LCDR3. In the present application, CDRs are defined by the IMGT numbering system, see Ehrenmann F, Kaas Q, Lefranc MP. IMGT / 3Dstructure-DB and IMGT / DomainGapAlign: a database and a tool for immunoglobulins or antibodies, T cell receptors, MHC, IgSF and MhcSF [J]. Nucleic acids research, 2009; 38(suppl_1): D301-D307.

[0427] The term "antibody" is not restricted by the method in which the antibody is produced. For example, it includes recombinant antibodies, monoclonal antibodies, and polyclonal antibodies. The antibody can be an antibody of different isotype, for example, an IgG (e.g., IgG1, IgG2, IgG3, or IgG4 subtype), IgA1, IgA2, IgD, IgE, or IgM antibody.

[0428] As used herein, the term "Fc", "Fc fragment" or "Fc fragment", also known as fragment crystallizable. Generally, the Fc fragment comprises the domain 2 (CH2) and the domain 3 (CH3) of the heavy chain constant region. In some embodiments of the present application, the Fc fragment is the Fc fragment of human IgG. In some embodiments of the present application, the Fc fragment of human IgG is the Fc fragment of human IgG1.

[0429] As used herein, the term "antigen-binding fragment" of an antibody refers to a polypeptide that comprises a fragment of a full-length antibody that retains the ability to specifically bind the same antigen bound by the full-length antibody, and / or competes with the full-length antibody for specific binding to the antigen, which is also referred to as an "antigen-binding portion." See generally, Fundamental Immunology, Ch. 7 (Paul, W., ed., 2nded. Raven Press, N.Y. (1989)), which is incorporated herein by reference in its entirety for all purposes. Antigen-binding fragments of an antibody can be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact antibodies. In some cases, antigen-binding fragments include Fab, Fab', F(ab')2, Fd, Fv, dAb, and complementarity determining region (CDR) fragments, single-chain antibodies (e.g., scFv), chimeric antibodies, diabodies, and polypeptides that contain at least a portion of an antibody that is sufficient to confer specific antigen binding to the polypeptide.

[0430] As used herein, the term "Fd fragment" means an antibody fragment consisting of the V H and C H 1 domains; the term "Fv fragment" means an antibody fragment consisting of the V L and V H domains of a single arm of an antibody; the term "dAb fragment" means an antibody fragment which consists of a V H domain (Ward et al., Nature 341 :544-546 (1989)); the term "Fab fragment" means an antibody fragment consisting of V L , V H , C L , and C H 1 domains; the term "F(ab')2 fragment" means an antibody fragment that comprises two Fab fragments linked by a disulfide bridge at the hinge region.

[0431] In some cases, an antigen-binding fragment of an antibody is a diabody, i.e., a bivalent antibody comprising V H and V L domains expressed on a single polypeptide chain, but using a linker that is too short to allow for pairing between the two domains on the same chain, thereby forcing the domains to pair with the complementary domains of another chain and creating two antigen binding sites (see, e.g., Holliger P. et al., Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993), and Poljak R.J. et al., Structure 2:1121-1123 (1994)).

[0432] As used herein, the terms "monoclonal antibody" and "monoclonal" refer to one antibody or one fragment of an antibody from a population of highly homogenous antibody molecules, i.e., a population of identical antibody molecules except for possible naturally occurring mutations that can arise during production. A monoclonal antibody has high specificity for a single epitope on an antigen. Polyclonal antibodies are in contrast to monoclonal antibodies and generally comprise at least 2 or more different antibodies, which generally recognize different epitopes on an antigen. Monoclonal antibodies can generally be obtained using the hybridoma technology first described by Kohler et al. (Kohler, G., Milstein, C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 1975; 256(5517): 495), but can also be obtained using recombinant DNA technology (see, e.g., U.S. Patent 4,816,567). G, Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity [J]. nature, 1975; 256(5517): 495), but can also be obtained using recombinant DNA technology (see, e.g., U.S. Patent 4,816,567).

[0433] As used herein, the term "humanized antibody" refers to an antibody or antibody fragment in which all or a portion of the CDR regions of a human immunoglobulin (recipient antibody) are replaced by CDR regions of a non-human antibody (donor antibody), which can be a non-human (e.g., mouse, rat, or rabbit) antibody having the desired specificity, affinity, or reactivity. Furthermore, some of the amino acid residues in the framework regions (FRs) of the recipient antibody can be replaced by corresponding amino acid residues of the non-human antibody, or by amino acid residues of other antibodies, to further refine or optimize antibody performance. For further details of humanized antibodies, see, e.g., Jones et al., Nature 1986; 321: 522 525; Reichmann et al., Nature, 1988; 332: 323 329; Presta, Curr. Op. Struct. Biol. 1992; 2: 593 596; and Clark, Immunol. Today 2000; 21: 397 402. In some instances, the antigen binding fragment of an antibody is a diabody, in which the VHand VLdomains are expressed in a single polypeptide chain, but using a linker that is too short to allow for pairing between the two domains on the same chain, thereby forcing the domains to pair with the complementary domains of another chain and creating two antigen binding sites (see, e.g., Holliger P. et al., Proc. Natl. Acad. Sci. USA 1993; 90: 6444 6448 and Poljak R.J. et al., Structure 1994; 2: 1121 1123).

[0434] As used herein, the term "single chain fragment variable (ScFv)" refers to a molecule comprising an antibody heavy chain variable region (VH) and an antibody light chain variable region (VL) linked by a linker. Wherein the VL and VH domains pair to form a monovalent molecule by a linker that enables it to be produced as a single polypeptide chain (see, e.g., Bird et al, Science 1988; 242:423-426 and Huston et al, Proc. Natl. Acad. Sci. USA 1988; 85:5879-5883). Such scFv molecules can have the general structure: NH2-VL-linker-VH-COOH or NH2-VH-linker-VL-COOH. Suitable prior art linkers consist of repeating GGGGS (SEQ ID NO: 15) amino acid sequences or variants thereof. For example, a linker having the amino acid sequence (GGGGS)4(SEQ ID NO: 16) can be used, but variants thereof can also be used (Holliger et al, Proc. Natl. Acad. Sci. USA 1993; 90:6444-6448). Other linkers useful in the present application are described by Alfthan et al, Protein Eng. 1995; 8:725-731, Choi et al, Eur. J. Immunol. 2001; 31:94-106, Hu et al, Cancer Res. 1996; 56:3055-3061, Kipriyanov et al, J. Mol. Biol. 1999; 293:41-56 and Roovers et al, Cancer Immunology, Immunotherapy, 2001, 50(1): 51-59.

[0435] As used herein, the terms "single domain antibody (VHH)", "nanobody" have the same meaning, referring to the variable region of the heavy chain of a cloned antibody, a single domain antibody (VHH) consisting of only one heavy chain variable region is constructed, which is the smallest antigen binding fragment with complete function. Usually, after obtaining an antibody naturally lacking a light chain and a heavy chain constant region 1 (CH1), the variable region of the heavy chain of the antibody is cloned to construct a single domain antibody (VHH) consisting of only one heavy chain variable region.

[0436] As used herein, the term "antibody moiety" refers to the antibody portion in an antibody-drug conjugate, which in certain specific protocols is linked to an intermediate linker moiety through a specific functional group, the antibody moiety can specifically bind to an antigen.

[0437] As used herein, the term "linker" refers to a moiety that links an antibody and a cytotoxin, and is classified as cleavable or non-cleavable. Cleavable linkers are mainly sensitive chemical bonds that can be cleaved by specific chemical species (e.g., glutathione, pH, etc.) or enzyme concentrations in the body, facilitating the cleavage of the linker from the drug, and are mainly connected by the form of hydrazone bond, disulfide bond, polypeptide. Non-cleavable linkers do not have built-in chemical bonds that can trigger cleavage, and need to be converted into amino acids by proteolytic mechanisms in cancer cells, thereby releasing cytotoxic drugs with linker and amino acid fragments, and are mainly connected by the form of thioether.

[0438] As used herein, the term "payload" refers to a cytotoxic molecule in an antibody-drug conjugate that is mainly responsible for performing the cell-killing function, which is coupled with an intermediate linker moiety, and enters tumor cells by internalization, and releases cytotoxic molecules under the action of lysosomes, thereby exerting an anti-tumor effect. The common target is DNA in the nucleus and tubulin in the cytoplasm.

[0439] As used herein, the term "DAR (drug-to-antibody ratio)" refers to the average number of small molecule drugs coupled to each antibody molecule. DAR is one of the important quality attributes of ADC. According to the chemical properties of the linker and the small molecule cytotoxin, as well as the coupling method (amino coupling, thiol coupling, site-specific coupling, etc.), common characterization analysis methods include ultraviolet-visible spectrophotometry, hydrophobic chromatography, reverse phase chromatography, and mass spectrometry (MS).

[0440] As used herein, the term "specifically binds" refers to a non-random binding reaction between two molecules, such as the reaction between an antibody and the antigen it targets. In certain embodiments, an antibody that specifically binds to (or has specificity for) an antigen means that the antibody binds to the antigen with an affinity (KD) of less than about 10 -5 M, for example, less than about 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 M, or 10 -10 M or less.

[0441] As used herein, the term "KD" refers to the dissociation equilibrium constant of a particular antibody-antigen interaction, which is used to describe the binding affinity between an antibody and an antigen. The smaller the equilibrium dissociation constant, the tighter the antibody-antigen binding, and the higher the affinity between the antibody and the antigen. Generally, an antibody binds to an antigen with an affinity (KD) of less than about 10 -5 M, for example, less than about 10 -6 M, 10 -7M, 10- 8 M, 10 - 9 M, or 10 -10 M or smaller dissociation equilibrium constant (KD) binds to the antigen. KD can be determined using methods known to those skilled in the art, for example, using a Fortebio molecular interaction instrument.

[0442] As used herein, the terms "monoclonal antibody" and "monoclonal" have the same meaning and are used interchangeably; the terms "polyclonal antibody" and "polyclonal" have the same meaning and are used interchangeably. Also in the present application, amino acids are generally represented by either the one-letter and three-letter abbreviations well-known in the art. For example, alanine can be represented by A or Ala.

[0443] As used herein, the term "pharmaceutically acceptable carrier and / or excipient" refers to a carrier and / or excipient that is compatible, in pharmacological and / or physiological terms, with the subject and active ingredient, which is well-known in the art (see, for example, Remington's Pharmaceutical Sciences. Edited by Gennaro AR, 19th ed. Pennsylvania: Mack Publishing Company, 1995), and includes, but is not limited to, pH adjusting agents, surfactants, adjuvants, ionic strength enhancers. For example, pH adjusting agents include, but are not limited to, phosphate buffers; surfactants include, but are not limited to, cationic, anionic or non-ionic surfactants, such as Tween-80; ionic strength enhancers include, but are not limited to, sodium chloride.

[0444] As used herein, the term "effective amount" refers to an amount sufficient to achieve or at least partially achieve a desired effect. For example, an effective amount for preventing a disease (e.g., a tumor) refers to an amount sufficient to prevent, arrest, or delay the onset of the disease (e.g., a tumor); an effective amount for treating a disease refers to an amount sufficient to cure or at least partially arrest the disease and its complications in an already afflicted patient. Determining such effective amounts is well within the capabilities of those skilled in the art. For example, an effective amount for therapeutic use will depend on the severity of the disease to be treated, the general state of the patient's own immune system, the general condition of the patient, such as age, body weight, and sex, the mode of administration of the drug, and other therapies concurrently administered, and the like.

[0445] In the present application, if not otherwise specified, the "first" (e.g., first active ingredient) and "second" (e.g., second active ingredient) are merely for the purpose of distinction in terms of reference or clarity in expression, and do not have the typical meaning of order.

[0446] Advantages of the invention

[0447] The present application achieves one or more of the following technical effects:

[0448] (1) The antibody conjugate drug prepared by the present application has good biological activity.

[0449] (2) The antibody conjugate drug prepared by the present application can effectively release the payload part.

[0450] (3) The anti-HER3 antibody-drug conjugate KA0013 of the present application can effectively bind to HER3.

[0451] (4) The anti-HER3 antibody-drug conjugate KA0013 of the present application has internalization activity.

[0452] (5) The anti-HER3 antibody-drug conjugate KA0013 of the present application has good stability in plasma.

[0453] (6) The drug combination of the present application has improved efficacy relative to the antibody conjugate drug.

[0454] (7) The drug combination of the present application has improved safety relative to the antibody conjugate drug. BRIEF DESCRIPTION OF DRAWINGS

[0455] Figure 1 : Anti-HER3 antibody-drug conjugate KA0013 and cell surface HER3 expressed on SK-BR-3 cells surface HER3 binding activity detection results.

[0456] Figure 2 : Anti-HER3 antibody-drug conjugate KA0013 internalization activity detection results in MDA-MB-453 cells expressing HER3 on the cell surface.

[0457] Figure 3 : Anti-HER3 antibody-drug conjugate KA0013 non-specific killing stability detection results caused by linker-payload part cleavage.

[0458] Figure 4 : Anti-HER3 antibody-drug conjugate KA0013 and KA0011 plasma stability detection results.

[0459] Figure 5 : In vitro detection of cathepsin B cleavage of KA0013 and KA0011 linker-payload efficiency.

[0460] Figure 6 : The efficacy of each group of administration regimens on CB-17 SCID mice HCC827 cell subcutaneous xenograft tumor models.

[0461] Figure 7Effects of different dosing regimens on body weight in CB-17SCID mouse HCC827 cell subcutaneous xenograft tumor models.

[0462] Figure 8 The efficacy of each dosing regimen on the CB-17SCID mouse SW620 cell subcutaneous xenograft model.

[0463] Figure 9 Effects of different dosing regimens on body weight in CB-17SCID mouse SW620 cell subcutaneous xenograft tumor models.

[0464] Figure 10 Effects of different dosing regimens on the blood concentration of ADC in CB-17SCID mouse SW620 cell subcutaneous xenograft tumor model.

[0465] Figure 11 The efficacy of each dosing regimen on the CB-17SCID mouse SW620 cell subcutaneous xenograft model.

[0466] Figure 12 Effects of different dosing regimens on body weight in CB-17SCID mouse SW620 cell subcutaneous xenograft tumor models.

[0467] Figure 13 Bystander killing activity assay of anti-HER3 antibody-drug conjugate.

[0468] Figure 14 : Detection of skin damage in hTROP2 mice by each dosing regimen. Detailed Implementation

[0469] The embodiments of the present invention will be described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are for illustrative purposes only and should not be considered as limiting the scope of the invention. Unless otherwise specified in the examples, conventional conditions or conditions recommended by the manufacturer are followed. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0470] In the following experimental examples of the present invention, the positive control ADC drug KA0011 was used. Its sequence information and preparation method are described in Chinese patent document CN 106163559 B.

[0471] In the following experimental examples of the present application, the isotype control antibodies used, IgG-DXd, IgG-KA0012 and IgG-KA0010, IgG-MMAE, the IgG (i.e. anti-HEL) component, and hlgG1 and anti-HEL are all antibodies targeting human anti-chicken egg lysosome (HEL), the variable region sequences of which are from Acierno et al. Affinity maturation increases the stability and plasticity of the Fv domain of anti-protein antibodies (Acierno et al. J Mol Biol. 2007; 374(1): 130-46.), and the constant region fragments are Ig gamma-1 chain C region, ACCESSION: P01857 as heavy chain constant region, Ig kappa chain C region, ACCESSION: P01834 as light chain constant region; IgG-DXd or IgG-KA0012, hlgG1 and anti-HEL are all prepared in the laboratory of Kangfang Biopharmaceutical Co., Ltd.

[0472] Preparation Example 1: Preparation of Anti-HER3 antibody Patritumab

[0473] Amino acid sequence of the heavy chain variable region of Patritumab:

[0474] QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQP PGKGLEWIGEINHSGSTNYNPSLKSRVTISVETSKNQFSLKLSSV TAADTAVYYCARDKWTWYFDLWGRGTLVTVSS (SEQ ID NO: 7)

[0475] Nucleic acid sequence encoding the heavy chain variable region of Patritumab:

[0476] CAGGTGCAGCTGCAGCAGTGGGGCGCCGGACTGCTGAAGCCATCCGAGACCCTGTCTCTGACATGTGCCGTGTACGGCGGCTCCTTTTCTGGCTACTATTGGAGCTGGATCAGGCAGCCCCCTGGCAAGGGACTGGAGTGGATCGGCGAGATCAACCACAGCGGCTCCACCAACTATAATCCCTCTCTGAAGAGCCGGGTGACCATCAGCGTGGAGACATCCAAGAATCAGTTCTCCCTGAAGCTGAGCTCCGTGACCGCAGCAGACACAGCCGTGTACTATTGCGCCCGGGACAAGTGGACCTGGTACTTTGATCTGTGGGGCAGAGGCACCCTGGTGACAGTGTCTAGC (SEQ ID NO: 8)

[0477] Amino acid sequence of the heavy chain variable region of Patritumab:

[0478] DIQMTQSPSSLSVTAEGVDRTI Y Y Y MD V AT TLTTVSDGKTYLN Y Y Y MD V AT GTI Y Y Y AMD V KGRFTISRDNAADTAIYYCARYDKGVPYFDYWGQGTLVTVSS (SEQ ID NO: 7)

[0479] Nucleic acid sequence encoding the heavy chain variable region of Patritumab:

[0480] GACATCGAGATGACCCAGTCCCCTGATTCTCTGGCCGTGAGCCTGGGAGAGAGGGCAACAATCAACTGTAGAAGCTCCCAGTCCGTGCTGTACTCTAGCTCCAACCGGAATTACCTGGCCTGGTATCAGCAGAATCCAGGCCAGCCCCCTAAGCTGCTGATCTATTGGGCCAGCACCAGGGAGTCCGGAGTGCCAGACCGCTTCTCCGGCTCTGGCAGCGGCACAGACTTCACCCTGACAATCTCTAGCCTGCAGGCCGAGGACGTGGCCGTGTACTATTGCCAGCAGTACTATAGCACCCCCAGGACATTTGGCCAGGGCACCAAGGTGGAGATCAAG (SEQ ID NO: 10)

[0481] The heavy chain constant region of Patritumab employs Ig gamma-1 chain C region; the light chain constant region employs Ig kappa chain C region.

[0482] The amino acid sequence of the heavy chain constant region of Patritumab:

[0483] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 11)

[0484] The nucleic acid sequence of the heavy chain constant region of Patritumab:

[0485] GACGGAGTGCACACATTTCCAGCCGTGCTGCAGAGCAGCGGACTGTATAGCCTGAGCAGCGTGGTGACCGTGCCTTCTTCTAGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGGGTGGAGCCCAAGTCTTGCGACAAGACCCACACTTGTCCTCCTTGTCCAGCCCCAGAGCTGCTGGGAGGACCAAGCGTGTTCCTGTTCCCTCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCAGAAGTGACTTGCGTGGTGGTGGACGTGTCTCACGAGGACCCCGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAGGTGCACAACGCTAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACAGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAATAAGGCCCTGCCAGCCCCTATCGAGAAGACCATCAGCAAGGCCAAGGGCCAGCCTAGAGAGCCTCAGGTGTACACCCTGCCTCCTTCTCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACTTGCCTCGTGAAGGGCTTCTACCCCAGCGATATTGCCGTCGAGTGGGAGTCTAACGGCCAGCCCGAGAACAACTACAAGACCACACCTCCAGTGCTGGATAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAAAGCCGCTGGCAGCAGGGCAACGTGTTTTCTTGCAGCGTGATGCACGAAGCCCTGCACAACCACTACACCCAGAAGAGCCTGAGCCTGTCTCCAGGCAAG (SEQ ID NO: 2)NO:12)

[0486] Amino acid sequence of the light chain constant region of Patritumab:

[0487] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 13)

[0488] Nucleic acid sequence of the light chain constant region of Patritumab:

[0489] CGTACGGTGGCAGCCCCATCTGTCTTCATTTTTCCCCCTAGTGACGAGCAGCTGAAATCCGGAACAGCCTCTGTGGTCTGTCTGCTGAACAATTTCTACCCTCGCGAAGCCAAGGTGCAGTGGAAAGTCGATAACGCTCTGCAGAGTGGCAATTCACAGGAGAGCGTGACTGAACAGGACTCCAAGGATTCTACCTATAGTCTGAGCTCCACTCTGACCCTGTCCAAAGCAGATTACGAAAAGCACAAAGTGTATGCCTGTGAGGTCACCCACCAGGGGCTGAGTTCTCCAGTCACCAAATCCTTCAACAGAGGCGAATGT (SEQ ID NO: 14)

[0490] The heavy chain cDNA and the light chain cDNA of Patritumab were cloned into pcDNA3.1 vector respectively to obtain the recombinant expression plasmid of the antibody Patritumab. The recombinant plasmid was transfected into 293F cells. The 293F cell culture fluid was purified and then detected.

[0491] The anti-HER3 monoclonal antibody Patritumab was prepared.

[0492] According to IMGT numbering system, the 6 CDRs of the anti-HER3 monoclonal antibody Patritumab are as follows:

[0493] HCDR1: GGSFSGYY (SEQ ID NO: 1)

[0494] HCDR2: INHSGST (SEQ ID NO: 2)

[0495] HCDR3: ARDKWTWYFDL (SEQ ID NO: 3)

[0496] LCDR1: QSVLYSSSNRNY (SEQ ID NO: 4)

[0497] LCDR2: WAS (SEQ ID NO: 5)

[0498] LCDR3: QQYYSTPRT (SEQ ID NO: 6).

[0499] Preparation Example 2: Preparation of Mc-AAA-DXd (KA0012)

[0500] Mc-AAA-DXd is (N-maleimidyl)-CH2CH2CH2CH2CH2-C(=0)-AAA-NH-CH2-0-CH2- C(=0)-(NH-DX). Wherein the CAS number of Dxd is: 1599440-33-1.

[0501] Mc-AAA-DXd is named as KA0012 in the present application, whose structural formula is shown as formula II below:

[0502]

[0503] Wherein the structure of N-maleimidyl is shown as formula III below:

[0504]

[0505] Wherein the structural formula of -(NH-DX) is shown as formula IV below:

[0506]

[0507] The synthesis steps of KA0012 are as follows: steps 1 to 10 below.

[0508] Step 1: Preparation of intermediate HM-1315_6B_2

[0509]

[0510] The starting material HM-1315_6B_1 (450.0 g, 1.44 mol, 1 eq) was suspended in DCM (4.5 L, 10 V) at 0-5 °C, and HOSU (182.9 g, 1.59 mol, 1.1 eq), EDCI (304.8 g, 1.59 mol, 1.1 eq) were added successively. After the addition was completed, the reaction solution was dissolved to be clear. The reaction was carried out at 10-20 °C for 3 h. Half-saturated brine (1.0 L) was added to the reaction solution, stirred, and then allowed to stand to separate. The organic phase was washed once with half-saturated brine (1.0 L) and twice with saturated brine (1.0 L x 2). After drying over anhydrous sodium sulfate, concentration was carried out to obtain 579.0 g of a white solid with a yield of 98%.

[0511] 1 H NMR (400 MHz, DMSO) δ 8.13 (d, J = 7.4 Hz, 1H), 7.89 (d, J = 7.5 Hz, 2H), 7.71 (t, J = 6.7 Hz, 2H), 7.42 (t, J = 7.3 Hz, 2H), 7.33 (t, J = 7.4 Hz, 2H), 4.52 (t, J = 7.4 Hz, 1H), 4.41-4.17 (m, 3H), 2.79 (d, J = 15.5 Hz, 4H), 2.59 (s, 1H), 1.46 (d, J = 7.4 Hz, 3H). LCMS [M+23]: 431.1.

[0512] Step 2: Preparation of intermediate HM-1315_1

[0513]

[0514] The starting material HM-1315_6B_2 (579.0 g, 1.42 mol, 1.0 eq) was dissolved in acetonitrile (2895 mL, 5 v), and glycine (117.2 g, 1.56 mol, 1.1 eq) and water (2895 mL, 5 v) were added. DIEA (275.3 g, 2.13 mol, 1.5 eq) was slowly added at -5-0 °C, and the reaction was carried out at 20-25 °C for 3 h. Ethyl acetate (1.0 L) was added to dilute the reaction solution, and dilute hydrochloric acid (0.5 N) was added to adjust the pH to 3-4. Ethyl acetate (500 mL) was added to extract and separate, and the aqueous phase was extracted twice with ethyl acetate (1.0 L x 2). The combined organic phase was washed once with saturated brine (1.5 L), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was slurried with methyl tert-butyl ether / ethyl acetate (2895 mL / 289.5 mL, 5 v / 0.5 v) for 2 h, filtered, and the filter cake was dried with an oil pump to obtain 447.2 g of a white powdery solid with a yield of 85%.

[0515] 1H NMR (400 MHz, DMSO) δ 8.12 (t, J = 5.5 Hz, 1H), 7.89 (d, J = 7.5 Hz, 2H), 7.73 (t, J = 7.1 Hz, 2H), 7.54 (d, J = 7.8 Hz, 1H), 7.37 (dt, J = 34.6, 7.1 Hz, 4H), 4.24 (dd, J = 15.6, 5.9 Hz, 3H), 4.13 - 4.05 (m, 1H), 3.75 (dd, J = 14.9, 5.7 Hz, 3H), 1.23 (d, J = 7.2 Hz, 3H). LCMS [M+l]: 369.1, [M+23]: 392.1.

[0516] Step 3: Preparation of intermediate HM-1315_2

[0517]

[0518] The starting material HM-1315_1 (474.0 g, 1.29 mol, 1.0 eq) was dissolved in super dry DMF (2370 mL, 5 v) under nitrogen protection, and lead tetraacetate (1143.9 g, 2.58 mol, 2.0 eq), anhydrous copper acetate (234.3 g, 1.29 mol, 1.0 eq) and acetic acid (170.5 g, 2.84 mol, 2.2 eq) were added, and the reaction temperature rose to about 50 °C. The reaction liquid was placed in an oil bath at 50-55 °C for 0.5 h. The reaction liquid was cooled, and ethyl acetate (2370 mL, 5 v) was added, followed by ice water (2370 mL, 5 v), and a black sticky solid was precipitated. The black insoluble material was filtered off with a Buchner funnel, and the filtrate was separated with a separatory funnel, and the water layer was extracted twice with ethyl acetate (2.0 L x 2), and the combined organic phase was washed twice with water (1.0 L x 2), and then washed three times with saturated brine (1.0 L x 3), dried over anhydrous sodium sulfate, filtered and concentrated, and the obtained crude product was slurried with methyl tert-butyl ether (2370 mL) at room temperature for 2 h, filtered, and the filter cake was dried with an oil pump to obtain 367.7 g of white solid product, with a yield of 74%.

[0519] 1 H NMR (400 MHz, DMSO) δ 8.90 (s, 1H), 7.89 (d, J = 7.5 Hz, 2H), 7.73 (t, J = 6.6 Hz, 2H), 7.59 (d, J = 7.6 Hz, 1H), 7.42 (t, J = 7.4 Hz, 2H), 7.33 (t, J = 7.4 Hz, 2H), 5.10 (d, J = 6.7 Hz, 2H), 4.25 (dd, J = 20.3, 6.5 Hz, 3H), 4.12 - 4.01 (m, 1H), 1.99 (s, 3H), 1.22 (d, J = 7.2 Hz, 3H). LCMS [M+23]: 405.0.

[0520] Step 4: Preparation of intermediate HM-1315_4

[0521]

[0522]

[0523] The starting material HM-1315_2 (326.0 g, 0.85 mol, 1.0 eq) was dissolved in THF (3260.0 mL, 10 v), and HM-1315_3 (211.9 g, 1.27 mol, 1.5 eq) was added. After the addition was complete, the reaction was cooled to 0-10 °C, and a solution of lithium hydroxide (24.4 g, 1.02 mol, 1.2 eq) in water (163.0 mL, 0.5 v) was slowly added. After the addition was complete, the reaction was allowed to naturally warm to room temperature (15-25 °C) for 1 hour. Ethyl acetate (1.5 L) and water (1.5 L) were added to the reaction solution, and the mixture was separated. The aqueous layer was extracted once more with ethyl acetate (1.0 L), and the combined organic phases were washed twice with saturated brine (1.0 L x 2), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product obtained after concentration was purified by column chromatography. The product was eluted with petroleum ether / ethyl acetate = 1 / 1, collected, concentrated, and dried with an oil pump to obtain 311.8 g of white solid product, with a yield of 74%.

[0524] 1 H NMR (400 MHz, DMSO) δ 8.74 (s, 1H), 7.89 (d, J = 7.5 Hz, 2H), 7.72 (t, J = 7.9 Hz, 2H), 7.59 (d, J = 7.3 Hz, 1H), 7.47 - 7.25 (m, 9H), 5.13 (s, 2H), 4.64 (d, J = 6.7 Hz, 2H), 4.31 - 4.18 (m, 3H), 4.15 (s, 2H), 4.07 - 3.95 (m, 1H), 1.23 (d, J = 7.2 Hz, 3H). LCMS [M+23]: 511.1.

[0525] Step 5: Preparation of intermediate HM-1315_7

[0526]

[0527]

[0528] The raw material HM-1315_4 (60.00 g, 122.82 mmol, 1.0 eq) was dissolved in DMAc (600.0 mL, 10 v), and cooled to 0-5 °C, and DBU (18.70 g, 122.82 mmol, 1.0 eq) was slowly added. After the addition was completed, the reaction was carried out at 0-5 °C for 1 hour. The reaction solution was cooled to -5-0 °C, and PPTS (30.86 g, 122.82 mmol, 1.0 eq), EDCI (23.54 g, 122.82 mmol, 1.0 eq), HOBT (16.59 g, 122.82 mmol, 1.0 eq) and HM-1315_6 (42.27 g, 110.54 mmol, 0.9 eq) were added to the reaction solution in turn, and the reaction was carried out at -5-0 °C for 16 hours. After the addition was completed, the reaction solution was added to n-hexane (1200 mL, 20 v) and allowed to stand to separate, and the DMAc layer was washed with n-hexane (1.2 L x 4) four times, and then poured into water (1800 mL, 30 v), and a large amount of white solid was precipitated. Filtration was performed using a Buchner funnel, and the filter cake was dried to obtain a white solid crude product. Acetonitrile (300.0 mL, 5 v) was added to the reaction bottle, and the white crude product obtained in the previous step was added, and the slurry was stirred for 1 hour, and then filtered, and the filter cake was dried by oil pump to obtain 56.2 g of white solid product, with a yield of 80%.

[0529] 1 H NMR (400 MHz, DMSO) δ 8.68 (t, J = 6.7 Hz, 1H), 7.99 (dd, J = 15.4, 7.2 Hz, 2H), 7.89 (d, J = 7.5 Hz, 2H), 7.72 (t, J = 7.0 Hz, 2H), 7.54 (d, J = 7.4 Hz, 1H), 7.46 - 7.28 (m, 9H), 5.14 (s, 2H), 4.63 (d, J = 7.7 Hz, 2H), 4.32 - 4.16 (m, 5H), 4.13 (s, 2H), 4.09 - 4.01 (m, 1H), 1.21 (dd, J = 7.0, 2.6 Hz, 9H). LCMS [M+23]: 654.3.

[0530] Step 6: Preparation of intermediate HM-1315_8

[0531]

[0532]

[0533] The starting material HM-1315_7 (2.59 g, 4.10 mmol, 1.0 eq) was suspended in THF (51.8 mL, 20 v) and water (5.2 mL, 2 v), cooled to 0-10 °C, 10% palladium on carbon (0.39 g, 0.15 w / w) was added, replaced with hydrogen gas for three times, and reacted at 0-10 °C for 16 hours. The palladium on carbon was removed by filtration, the filter cake was rinsed with acetonitrile / water (10 mL / 10 mL) again, 2-methyltetrahydrofuran (50 mL) was added to the filtrate to extract, the organic phase was washed with water (50 mL) and saturated brine (50 mL) once respectively, dried over anhydrous sodium sulfate, filtered and concentrated to give 2.0 g of crude product as a white solid. The crude product (2.0 g, 3.70 mmol, 1.0 eq) was suspended in dichloromethane (20.0 mL, 10 v), DBU (0.56 g, 3.70 mmol, 1.0 eq) was added dropwise slowly at 20-25 °C, the reaction solution was partially dissolved after the addition was completed, and reacted at 20-25 °C for 16 hours, during which white solid was precipitated. Methyl tert-butyl ether (40.0 mL, 20 v) was added to the reaction solution, and more white solid was precipitated. The reaction solution was filtered, and the filter cake was transferred to a single-neck flask, acetonitrile (20.0 mL) was added, and the slurry was stirred for 2 h, filtered, and the filter cake was dried by oil pump to give 0.5 g of product as a white solid, with a yield of 38%.

[0534] 1 H NMR (400 MHz, D2O) δ 4.73 (q, J = 10.8 Hz, 1H), 4.43-4.21 (m, 1H), 4.04-3.82 (m, 1H), 1.47 (d, J = 7.0 Hz, 2H), 1.42 (d, J = 7.2 Hz, 3H). LCMS [M+l]: 319.1.

[0535] Step 7: Preparation of intermediate HM-1315_9

[0536]

[0537] The starting material HM-1315_8 (300 mg, 0.94 mmol, 1.0 eq) was dissolved in acetonitrile (3.0 mL, 10 v) and water (4.5 mL, 15 v), HM-297D_9 (290 mg, 0.94 mmol, 1.0 eq) was added, cooled to 0-10 °C, DIPEA (121 mg, 0.94 mmol, 1.0 eq) was added, and reacted at 0-10 °C for 16 hours to give the product (290 mg, 60% yield) as a white solid.

[0538] 1H NMR (400 MHz, DMSO) d 8.61 (t, J = 6.7 Hz, 1H), 7.97 (dd, J = 7.1, 3.4 Hz, 2H), 7.91 (d, J = 7.3 Hz, 1H), 7.00 (s, 2H), 4.66 - 4.52 (m, 2H), 4.29 - 4.12 (m, 3H), 3.95 (s, 2H), 3.44 - 3.35 (m, 5H), 2.08 (t, J = 7.4 Hz, 2H), 1.54 - 1.40 (m, 4H), 1.28 - 1.10 (m, 11H). LCMS [M+23]: 534.3.

[0539] Step 8: Preparation of intermediate HM-582_9

[0540]

[0541] Compound HM-582_7 (10 g, 39.98 mmol, 1.0 eq), compound HM-582_8 (10 g, 38.01 mmol, 0.95 eq), p-toluene sulfonic acid pyridine salt (6 g, 23.88 mmol, 0.60 eq) were added into a three-neck flask, toluene (500 mL) was added, the reaction was carried out at 130-140 °C for 48 hours, the reaction solution was cooled, filtered, washed with methyl tert-butyl ether (100 mL), the solid was collected and oven dried to give the product (18.4 g, 96.5%).

[0542] 1 H NMR (DMSO) d: 8.43 - 8.55 (m, 1H), 7.80 (d, J = 10.9 Hz, 1H), 7.31 (d, J = 3.8 Hz, 1H), 6.55 (br s, 1H), 5.52 - 5.57 (m, 1H), 5.42 (s, 2H), 5.16 - 5.20 (m, 2H), 3.14 - 3.18 (m, 2H), 2.39 (s, 3H), 2.11 - 2.16 (m, 2H), 1.80 - 1.92 (m, 5H), 0.86 - 0.89 (m, 3H). LCMS [M+1] 478.2.

[0543] Step 9: Preparation of intermediate HM-582_10

[0544]

[0545] Compound HM-582_9 (30 g) was suspended in water (600 mL), methanesulfonic acid (300 mL) was added slowly, the solid dissolved and there was an exothermic phenomenon, after nitrogen replacement, heated to 112 °C for 7 h, cooled to room temperature, filtered, the filter cake was washed with water (100 mL). The filtrate was diluted with ethanol (4 L), the solid precipitated, stirred at room temperature for 20 min, filtered, dried under suction, the crude was suspended in ethanol / water = 4: 1 (1 L), heated to reflux for 2 h, cooled to room temperature, filtered, the solid was washed with a small amount of ethanol and dried under suction, after lyophilization, irinotecan methanesulfonate (16.7 g, 50%) was obtained.

[0546] 1 H NMR (DMSO) δ 8.41-8.50 (m, 3H), 7.86 (d, J = 10.8 Hz, 1H), 7.33 (s, 1H), 5.66-5.74 (m, 1H), 5.37-5.44 (m, 2H), 5.10 (s, 1H), 3.27-3.32 (m, 1H), 3.08-3.17 (m, 1H), 2.41 (s, 3H), 2.31 (s, 3H), 2.17-2.24 (m, 1H), 1.80-1.95 (m, 2H), 0.88 (t, J = 7.3 Hz, 3H). LCMS [M+1] 436.1.

[0547] Step 10: Preparation of HM-1315_10 (KA0012)

[0548]

[0549] The starting material HM-1315_9 (150 mg, 0.29 mmol, 1.0 eq) was dissolved in DMAc (3.0 mL, 20v), HM-582_10 (156 mg, 0.29 mmol, 1.0 eq) was added, cooled to 0-10 °C, HATU (121 mg, 0.32 mmol, 1.0 eq) was added, 2,4,6-trimethylpyridine (70 mg, 0.58 mmol, 2.0 eq) was added dropwise, and the reaction was carried out at 0-10 °C for 16 h. LCMS showed that the starting material was completely converted, and the product (135 mg, 50% yield) was obtained as a light yellow solid by preparation separation.

[0550] 1H NMR (400 MHz, DMSO) δ 8.64 (t, J = 6.6 Hz, 1H), 8.47 (d, J = 8.8 Hz, 1H), 7.95 (dd, J = 9.3, 7.3 Hz, 2H), 7.87 (d, J = 7.2 Hz, 1H), 7.77 (d, J = 11.0 Hz, 1H), 7.30 (s, 1H), 6.99 (s, 2H), 6.51 (s, 1H), 5.60 (d, J = 8.0 Hz, 1H), 5.42 (s, 2H), 5.18 (s, 2H), 4.61 (dt, J = 10.1, 3.3 Hz, 2H), 4.17 (dt, J = 14.1, 7.1 Hz, 3H), 3.98 (s, 2H), 3.38 (s, 1H), 3.16 (dd, J = 14.1, 6.4 Hz, 2H), 2.38 (s, 3H), 2.18 (d, J = 5.4 Hz, 2H), 2.07 (t, J = 7.3 Hz, 2H), 1.85 (dd, J = 15.0, 7.4 Hz, 2H), 1.51 - 1.39 (m, 4H), 1.17 (dt, J = 14.5, 7.4 Hz, 11H), 0.86 (t, J = 7.3 Hz, 3H). LCMS [M+l]: 929.3, [M+23]: 952.3.

[0551] Preparation Example 3: Preparation of Anti-HER3 antibody-drug conjugate KA0013

[0552] The structures of KA0013 and KA0011 are shown below in Table 1.

[0553] Table 1

[0554]

[0555] wherein GGFG is represented as SEQ ID NO: 20.

[0556] An antibody-drug conjugate is obtained by forming a thioether bond in the disulfide bond site present in the hinge region of an anti-HER3 antibody, wherein the anti-HER3 antibody is treated under reducing conditions and then reacted with a compound (e.g., Formula II) to form the antibody-drug conjugate. Specifically, the reaction of the compound with the antibody having a thiol group can produce the antibody-drug conjugate.

[0557] An antibody having a thiol group can be obtained using a method known in the art (Hermanson, G. T, Bioconjugate Techniques, pp. 56-136, 456-493, Academic Press (1996)). Examples include: reaction of Traut's reagent with the amino group of the antibody; reaction of N-succinimidyl S-acetylthioalkanoate with the amino group of the antibody followed by reaction with hydroxylamine; reaction with a reducing agent after reaction with N-succinimidyl 3-(pyridyldithio)propionate; reaction of the antibody with a reducing agent such as dithiothreitol, 2-mercaptoethanol, and tris(2-carboxyethyl)phosphine hydrochloride (TCEP) to reduce the disulfide bond of the hinge portion within the antibody to form a thiol group, but it is not limited thereto.

[0558] Specifically, an antibody in which the disulfide bond of the hinge portion within the antibody is partially or completely reduced can be obtained by using 0.3 to 10 molar equivalents of TCEP as a reducing agent, in relation to 1 disulfide bond of the hinge portion within the antibody, and reacting it with the antibody in a buffer containing a chelating agent. Examples of the chelating agent include ethylenediaminetetraacetic acid (EDTA) and diethylenetriaminepentaacetic acid (DTPA).

[0559] It can be at a concentration of 1 mM to 20 mM. Examples of the buffer that can be used include a sodium phosphate solution, a sodium borate solution, or a sodium acetate solution. Specifically, an antibody having a thiol group that is partially or completely reduced can be obtained by reacting the antibody with TCEP at 4°C to 37°C for 1 to 4 hours.

[0560] Meanwhile, by performing a reaction in which a thiol group is added to the drug-linker moiety, the drug-linker moiety can be coupled through a thioether bond.

[0561] In relation to 1 antibody having a thiol group, 2 to 20 molar equivalents of the compound can be used to produce an antibody-drug conjugate in which 2 to 8 drug molecules are coupled per 1 antibody. Specifically, it is sufficient to add a solution in which the compound is dissolved to a buffer containing the antibody having a thiol group to perform the reaction. Here, examples of the buffer that can be used include a sodium acetate solution, a sodium phosphate, and a sodium borate. The pH of the reaction is 5 to 9, and more preferably the reaction is performed at around pH 7. Examples of the solvent used to dissolve the compound (1) include organic solvents such as dimethyl sulfoxide (DMSO), dimethylformamide (DMF), dimethylacetamide (DMA), and N-methyl-2-pyrrolidone (NMP).

[0562] The solution of the organic solvent in which the compound is dissolved can be added to the buffer containing the antibody having a thiol group at 1 to 20% v / v to perform the reaction. The reaction temperature is 0 to 37°C, more preferably 10 to 25°C, and the reaction time is 0.5 to 2 hours.

[0563] After the conjugation, the manufactured antibody-drug conjugate can be subjected to buffer exchange, measurement of the antibody concentration and average number of drugs per antibody molecule to identify the antibody-drug conjugate.

[0564] Common Operation A: Measurement of Antibody Concentration

[0565] The measurement of the antibody concentration was performed using a UV detector (Nanodrop 1000, Thermo Fisher Scientific Inc.) according to the method specified by the manufacturer. At this time, the known calculation method (Protein Science, 1995, Vol. 4, 2411-2423) can be used to calculate the theoretical absorbance coefficient at 280 nm according to the amino acid sequence of the antibody, such as 1.768 mL·mg -1 ·cm -1 of the absorbance coefficient at 280 nm according to the amino acid sequence of Patritumab as a presumption value.

[0566] Common Operation B: Buffer Exchange of Antibody-Drug Conjugate

[0567] The antibody-drug conjugate was subjected to ultrafiltration concentration and dialysis using an ultrafiltration membrane pack (Millipore P3C030C00, 30 kDa) according to the method specified by the manufacturer, and the dialysis was performed 10 times.

[0568] Common Operation C: Measurement of Average Number of Drugs per Antibody Molecule in Antibody-Drug Conjugate

[0569] The average number of drugs per antibody molecule in the antibody-drug conjugate was measured using liquid chromatography-mass spectrometry:

[0570] The sample was diluted with water to 2 mg / mL, 50 μL of the diluted sample was taken, 1 μL of 0.5 M TCEP was added, and the reaction was performed at room temperature for 30 min, and then the sample was prepared for injection. The chromatographic column was MabPac RP (2.1 x 50 mm), the mobile phase was mobile phase A-0.1% formic acid aqueous solution, mobile phase B-0.1% formic acid acetonitrile solution, the injection amount was 2 μg, the column temperature was 80°C, and the detection wavelength was 280 nm.

[0571] The liquid phase gradient was as shown in Table 2 below.

[0572] Table 2

[0573]

[0574] DAR values are calculated from the individual signal values.

[0575]

[0576] In the above formula, LCn represents the signal value of a light chain coupled with n linkers-payloads, and HCn represents the signal value of a heavy chain coupled with n linkers-payloads.

[0577] A specific conjugation example is as follows.

[0578] Reduction of antibody: Partritumab was diluted to about 10 mg / mL using phosphate buffer PB / EDTA (15 mM Na2HPO4, 5 mM NaH2PO4, 5 mM EDTA-2Na, pH 7.0). The antibody solution (100 mL) was added to a beaker at room temperature, followed by the addition of 10 mM TCEP aqueous solution (6.8 mL; 10.0 equivalents relative to one antibody molecule), and stirring using a magnetic stirrer, and incubated at room temperature for 3 hours to reduce the disulfide bond of the hinge part of the antibody.

[0579] Conjugation of antibody with drug linker: 10 mM KA0012-containing DMSO solution (7.48 mL; 11.0 equivalents relative to one antibody molecule) was slowly added thereto. Stirring at room temperature for 1 hour to conjugate the drug linker with the antibody.

[0580] Purification: ultrafiltration concentration exchange was performed using an ultrafiltration membrane bag, and the buffer was replaced into a formulation buffer while removing unconjugated drug linkers and other low molecular weight reagents, and the solution was concentrated. The resulting purified solution was subjected to sterilization filtration to obtain a solution containing antibody-drug conjugate KA0013.

[0581] Measurement of antibody concentration using Common Operation A: 23.64 mg / mL, measurement of average number of drug molecules conjugated per one antibody molecule (n) in KA0013 using Common Operation C: 7.87.

[0582] Preparation Example 4: Preparation of control drug

[0583] KA0010: Mc-GGFG-DXd, linker-payload as a control.

[0584] The structural formula of the control KA0010 is shown in the following Formula V:

[0585]

[0586]

[0587] KA0011: Patritumab-MC-GGFG-DXd.

[0588] IgG-KA0010: IgG-Mc-GGFG-DXd, abbreviated as IgG-DXd.

[0589] IgG-KA0012: IgG-Mc-AAA-DXd.

[0590] The preparation method of KA0010 and KA0011 refers to the relevant description in Chinese patent document CN 106163559B.

[0591] IgG-KA0010 and IgG-KA0012 are both prepared by using the common coupling method for thiol coupling, and the specific preparation method refers to the preparation method of KA0013 above, and also refers to the relevant description in Chinese patent document CN 106163559B.

[0592] The average drug coupling number (n) of each antibody molecule in KA0011, IgG-KA0010, and IgG-KA0012: 8.

[0593] Preparation Example 5: Preparation of anti-Trop2 antibody hRS7 (G1TM)

[0594] Amino acid sequence of the heavy chain variable region of hRS7 (G1TM):

[0595] QVQLQQSGSELKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYTDDFKGRFAFSLDTSVSTAYLQISSLKADDTAVYFCARGGFGSSYWYFDVWGQGSLVTVSS (SEQ ID NO: 17)

[0596] Amino acid sequence of the light chain variable region of hRS7 (G1TM):

[0597] DIQLTQSPSSLSASVGDRVSITCKASQDVSIAVAWYQQKPGKAPKLLIYSASYRYTGVPDRFSGSGSGTDFTLTISSLQPEDFAVYYCQQHYITPLTFGAGTKVEIK (SEQ ID NO: 18)

[0598] The heavy chain constant region of hRS7 (G1TM) adopts Ig gamma-1 chain C region; and the light chain constant region adopts Ig kappa chain C region.

[0599] The amino acid sequence of the heavy chain constant region of hRS7(G1TM):

[0600] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 19)

[0601] The amino acid sequence of the light chain constant region of hRS7(G1TM):

[0602] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 13)

[0603] The heavy chain cDNA and the light chain cDNA of hRS7(G1TM) were cloned into pcDNA3.1 vector respectively to obtain the recombinant expression plasmid of antibody hRS7(G1TM). The recombinant plasmid was transfected into 293F cells. The 293F cell culture fluid was purified and then detected.

[0604] The anti-Trop2 monoclonal antibody hRS7(G1TM) was prepared.

[0605] According to the IMGT numbering system, the 6 CDRs of the anti-Trop2 monoclonal antibody hRS7(G1TM) are as follows:

[0606] HCDR1: GYTFTNYG (SEQ ID NO: 21)

[0607] HCDR2: INTYTGEP (SEQ ID NO: 22)

[0608] HCDR3: ARGGFGSSYWYFDV (SEQ ID NO: 23)

[0609] LCDR1: QDVSIA (SEQ ID NO: 24)

[0610] LCDR2: SAS (SEQ ID NO: 25)

[0611] LCDR3: QQHYITPLT (SEQ ID NO: 26).

[0612] Preparation Example 6: Preparation of Anti-Trop2-MMAE The structure of Anti-Trop2-MMAE is shown in Table 3 below.

[0613] Table 3

[0614] Nomenclature Structure Anti-Trop2-MMAE hRS7(G1TM)-mc-vc-PAB-MMAE

[0615] Mc-vc-PAB-MMAE, also known as VcMMAE, is a MMAE derivative with a valine-citrulline (Vc) linker for the synthesis of antibody-drug conjugates (ADCs). Among them, Mc-VC-PAB is a cleavable ADC linker, and MMAE (Monomethyl auristatin E) is a synthetic antitumor drug and also a microtubulin inhibitor, which can be effectively released from VcMMAE in vitro and exert cytotoxic activity.

[0616] The structure of mc-vc-PAB-MMAE is shown in the following formula VI:

[0617]

[0618] An antibody-drug conjugate is obtained by forming a thioether bond in the disulfide bond site present in the hinge portion of the anti-Trop2 antibody, wherein the anti-Trop2 antibody is treated under reducing conditions, and then reacted with a compound (such as Formula II) to form an antibody-drug conjugate. Specifically, the reaction of the compound with the antibody having a thiol group can produce an antibody-drug conjugate.

[0619] The antibody having a thiol group can be obtained using a method known in the art (Hermanson, G. T, Bioconjugate Techniques, pp. 56-136, 456-493, Academic Press (1996)). Examples include: reaction of Traut's reagent with the amino group of the antibody; reaction of N-succinimidyl S-acetylthioalkanoate with the amino group of the antibody followed by reaction with hydroxylamine; reaction with a reducing agent after reaction with N-succinimidyl 3-(pyridyldithio)propionate; reaction of the antibody with a reducing agent such as dithiothreitol, 2-mercaptoethanol, and tris(2-carboxyethyl)phosphine hydrochloride (TCEP) to reduce the disulfide bond of the hinge portion within the antibody to form a thiol group, but it is not limited thereto.

[0620] Specifically, an antibody in which the disulfide bond of the hinge portion within the antibody is partially or completely reduced can be obtained by using 0.3 to 10 molar equivalents of TCEP as a reducing agent, reacting it with the antibody in a buffer, with respect to 1 disulfide bond of the hinge portion within the antibody.

[0621] It can be at a concentration of 1 mM to 20 mM. Examples of the buffer that can be used include a phosphate buffer system, a histidine-histidine hydrochloride buffer system, or a citric acid-sodium citrate buffer system. Specifically, an antibody having a partially or completely reduced thiol group can be obtained by reacting the antibody with TCEP at 4°C to 37°C for 1 to 24 hours.

[0622] Meanwhile, by performing a reaction in which a thiol group is added to the drug-linker moiety, the drug-linker moiety can be coupled through a sulfide bond.

[0623] With respect to 1 antibody having a thiol group, 2 to 20 molar equivalents of the compound can be used to produce an antibody-drug conjugate in which 2 to 8 drug molecules are coupled per 1 antibody. Specifically, it is sufficient to add a solution in which the compound is dissolved to a buffer containing the antibody having a thiol group to perform the reaction. Here, examples of the buffer that can be used include a phosphate buffer system, a histidine-histidine hydrochloride buffer system, or a citric acid-sodium citrate buffer system. The pH of the reaction is 5 to 9, and more preferably the reaction is performed at around pH 7. Examples of the solvent used to dissolve the compound (1) include organic solvents such as dimethyl sulfoxide (DMSO), dimethylformamide (DMF), dimethylacetamide (DMA), and N-methyl-2-pyrrolidone (NMP).

[0624] The solution of the organic solvent in which the compound is dissolved can be added to the buffer containing the antibody having a thiol group in 1 to 20% v / v to perform the reaction. The reaction temperature is 0 to 37°C, more preferably 10 to 25°C, and the reaction time is 0.5 to 2 hours.

[0625] After the conjugation, the manufactured antibody-drug conjugate can be subjected to buffer exchange, measurement of the antibody concentration and average number of drugs per antibody molecule to identify the antibody-drug conjugate.

[0626] Common Operation A: Measurement of Antibody Concentration

[0627] The measurement of the antibody concentration was performed using a UV detector (Nanodrop 1000, Thermo Fisher Scientific Inc.) according to the method prescribed by the manufacturer. At this time, the known calculation method (Protein Science, 1995, Vol. 4, 2411-2423) can be used to calculate the theoretical absorbance coefficient at 280 nm according to the amino acid sequence of the antibody, such as 1.560 mL·mg -1 ·cm -1 of the absorbance coefficient at 280 nm according to the amino acid sequence of hRS7 (G1TM) monoclonal antibody was used as a presumptive value.

[0628] Common Operation B: Buffer Exchange of Antibody-Drug Conjugate

[0629] The antibody-drug conjugate was subjected to ultrafiltration concentration and dialysis using an ultrafiltration membrane pack (Sartorius Hysard, 30 kDa) according to the method prescribed by the manufacturer, and the dialysis was performed 10 times.

[0630] Common Operation C: Measurement of Average Number of Drugs per Antibody Molecule in Antibody-Drug Conjugate

[0631] The average number of drugs per antibody molecule in the antibody-drug conjugate was measured using liquid chromatography-mass spectrometry:

[0632] The sample was diluted with water to 2 mg / mL, 50 μL of the diluted sample was taken, 1 μL of 0.5 M TCEP was added, and the reaction was performed at room temperature for 30 min, and then the sample was prepared for injection. The chromatographic column was MabPac RP (2.1 x 50 mm), the mobile phase was mobile phase A-0.1% formic acid aqueous solution, mobile phase B-0.1% formic acid acetonitrile solution, the injection amount was 2 μg, the column temperature was 80°C, and the detection wavelength was 280 nm.

[0633] The liquid phase gradient was as shown in Table 4 below.

[0634] Table 4

[0635]

[0636] DAR values are calculated from the values of the individual component signals.

[0637]

[0638] In the above formula, LCn represents the signal value of a light chain coupled with n linkers-payloads, and HCn represents the signal value of a heavy chain coupled with n linkers-payloads.

[0639] A specific coupling example is as follows.

[0640] Reduction of the antibody: 10 mM TCEP aqueous solution (10.0 equivalents relative to one antibody molecule) was added to the anti-Trop2 antibody, and stirring was performed using a magnetic stirrer, and incubation was performed at room temperature for 3 hours to reduce the disulfide bond of the hinge portion of the antibody.

[0641] Coupling of the antibody with the drug linker: 10 mM VcMMAE (mc-vc-PAB-MMAE, purchased from MedChemExpress, item number: HY-15575) in DMSO solution (5.0 equivalents relative to one antibody molecule) was slowly added thereto. Stirring was performed at room temperature for 1 hour to couple the drug linker with the antibody.

[0642] Purification: ultrafiltration concentration exchange was performed using an ultrafiltration membrane bag, and the buffer was replaced with a formulation buffer while removing the un-coupled drug linker and other low molecular weight reagents, and the solution was concentrated. The obtained purified solution was subjected to sterilization filtration to obtain a solution containing the antibody-drug conjugate Anti-Trop2-MMAE.

[0643] Measurement of the antibody concentration using Common Operation A: 8.15 mg / mL, measurement of the average number of drugs coupled per antibody molecule (n) in Anti-Trop2-MMAE using Common Operation C: 4.

[0644] IgG-MMAE: IgG-mc-vc-PAB-MMAE, the specific preparation method of which is described above. The average number of drugs coupled per antibody molecule (n) in IgG-MMAE: 4.

[0645] Example 1: FACS detection of the binding activity of Anti-HER3 antibody-drug conjugate KA0013 to HER3 on the surface of SK-BR-3 cells Figure 1

[0646] 1. Experimental drug

[0647] KA0013,

[0648] HER3 target monoclonal antibody Patritumab (prepared according to Preparation Example 1).

[0649] KA0011.

[0650] 2. Experimental method

[0651] SK-BR-3 cells (purchased from Shanghai Institute of Life Sciences, Chinese Academy of Sciences) were routinely collected, washed once with an appropriate amount of PBS, counted and the cell viability was determined; 3 x 10 5 The cell suspension was added to a 96-well plate at 3 x 10 50 The cell pellet was resuspended and incubated on ice for 40 minutes; 1% PBSA (PBS + 1% BSA) was added, centrifuged at 1000 x g for 5 minutes, and the supernatant was discarded; the washing was repeated twice; 100 μL of 300-fold diluted fluorescent antibody Mouse Anti-Human IgG Fc-Alexa Fluor 647 (SouthernBiotech, Cat. 9040-31) was added, the cell pellet was resuspended, and incubated on ice for 30 minutes in the dark; 1% PBSA was added, centrifuged at 1000 x g for 5 minutes, and the supernatant was discarded; the washing was repeated twice; 200 μL of 1% PBSA was added, and the cell pellet was resuspended for machine detection.

[0652] 3. Experimental results

[0653] The results are shown in Table 1. Example 2: FACS detection of the internalization activity of Anti-HER3 antibody-drug conjugate KA0013 on MDA-MB-453 cells

[0654] The results show that, under the same experimental conditions, the EC 50 of KA0013, KA0011, and HER3 target monoclonal antibody Patritumab binding to SK-BR-3 cells were 15.36 nM, 12.89 nM, and 8.481 nM, respectively.

[0655] The results show that, under the same experimental conditions, KA0013, KA0011, and Patritumab all have the activity of effectively binding to SK-BR-3 cells expressing HER3.

[0656] Figure 2

[0657] 1. Experimental drug

[0658] KA0013,

[0659] IgG-DXd,

[0660] KA0012,

[0661] KA0011.

[0662] 2. Experimental method

[0663] MDA-MB-453 cells (purchased from Nanjing Kebai Biotechnology Co., Ltd., item number: CBP60386) were collected, counted and the cell viability was determined; 1 x 10 5 The cell suspension was added to a 96-well low adsorption plate at 1 x 10

[0664] 3. Experimental results

[0665] The results are shown in Table 1. Example 3: Measurement of the non-specific killing activity of the linker-payload portion of Anti-HER3 antibody-drug conjugate

[0666] The results show that, under the same experimental conditions, KA0013 and KA0011 can be endocytosed in MDA-MB-453, and the endocytosis activity of the two is significantly different compared with the isotype control IgG-DXd (i.e. IgG-Mc-GGFG-DXd), produced by Zhongshan Kangfang Biotechnology Co., Ltd., batch number: 20230306).

[0667] Figure 3 Example 4: Measurement of the plasma stability of Anti-HER3 antibody-drug conjugate KA0013

[0668] 1. Experimental drug

[0669] IgG-KA0010,

[0670] IgG-KA0012.

[0671] 2. Experimental method

[0672] ​Non-specific killing activity detection was performed on the linker-payload part Mc-AAA-DXD (named KA0012) of the anti-HER3 antibody-drug conjugate KA0013 and the linker-payload part Mc-GGFG-DXD (named KA0010) of the control KA0011.

[0673] After collecting human venous blood, the heparin sodium anticoagulation tube was gently mixed and inverted, centrifuged at 3500 rpm for 10 minutes at room temperature, and the plasma layer was taken for experiment. MDA-MB-453 cells were routinely collected, counted and the cell viability was determined, and the cell density was adjusted to 2.5×10 4 After collecting human venous blood, the heparin sodium anticoagulation tube was gently mixed and inverted, centrifuged at 3500 rpm for 10 minutes at room temperature, and the plasma layer was taken for experiment. MDA-MB-453 cells were routinely collected, counted and the cell viability was determined, and the cell density was adjusted to 2.5×10

[0674] 3. Experimental results

[0675] The results are shown in Table 1. Time (min)

[0676] The results show that under the same experimental conditions, compared with the isotype control (IgG-KA0010, produced by Zhongshan Kangfang Biotechnology Co., Ltd., batch number: 20230306), the non-specific killing of IgG-KA0012 is weaker, indicating that Mc-AAA-DXd is more stable than Mc-GGFG-DXd.

[0677] Flow rate (mL / min)

[0678] 1. Experimental drug

[0679] KA0013,

[0680] KA0011.

[0681] 2. Experimental method

[0682] ​After collecting human venous blood, gently mix the heparin sodium anticoagulation tube up and down, centrifuge at 3500 rpm for 10 minutes at room temperature, and the supernatant is the plasma. Dilute the ADC sample with human plasma (working concentration is 100 μg / mL), incubate at 37°C in a 5% CO2 incubator for 0h, 24h, 48h, 72h, 96h, 139h, after incubation, take out 150 μL sample and freeze in liquid nitrogen, after collecting all samples, detect the change of free DXd in plasma by mass spectrometry.

[0683] Mass spectrometry sample preparation: take the above sample, mix with acetonitrile 1:1 to precipitate protein, centrifuge at 12000 rpm for 3 min, take 10 μL supernatant for mass spectrometry detection. Chromatographic column: MabPac RP 2.1x50mm 4μm; mobile phase A: 0.1% formic acid water; mobile phase B: 0.1% formic acid acetonitrile; column temperature: 30°C;

[0684] The gradient is as follows in Table 5.

[0685] Table 5

[0686] Figure 4 Example 5: In vitro detection of the cleavage efficiency of Cathepsin B on different linkers-payloads A(％) B(％) 0 0.4 95 5 1 0.4 95 5 5 0.4 10 90 6 0.4 10 90 6.5 0.4 95 5 7 0.4 10 90 8 0.4 10 90 8.1 0.4 95 5 10 0.4 95 5

[0687] 3. Experimental results

[0688] The results are shown in Time (min) .

[0689] The results show that under the same experimental conditions, the amount of free DXd in the plasma of the KA0013 group is lower than that of the KA0011 group, indicating that the stability of KA0013 in plasma is better than that of KA0011.

[0690] Flow rate (mL / min)

[0691] 1. Experimental sample:

[0692] KA0010,

[0693] KA0012.

[0694] 2. Experimental method

[0695] Dilute PBS (20-fold solution) (Shenguo, Cat.: B548117) 20-fold to obtain PBS working solution, dilute Cathepsin B (CTSB) (Sinobio, Cat.: 10483-H08H) and linker-payload with PBS working solution; mix 150 μL CTSB (working concentration is 3 nM) with equal volume of linker-payload (working concentration is 10 μM), incubate in a 37°C, 5% CO2 incubator for 0h, 0.5h, 1h, 2h, 4h; after incubation, add Halt Protease Inhibitor Cocktail (Thermo, Cat.: 87786) to stop the reaction, and then take 10 μL sample for mass spectrometry detection. TMProtease Inhibitor Cocktail, EDTA-Free (30X) (Thermofisher, Cat.: 87785) to terminate the reaction.

[0696] The sample after enzymolysis was directly loaded for mass spectrometry detection. The chromatographic column was MabPac RP 2.1x50mm 4um; the mobile phase A was 0.1% formic acid water; the mobile phase B was 0.1% formic acid acetonitrile; the sample loading amount was 10uM sample injection 1uL; the column temperature was 30°C.

[0697] The gradient was as shown in Table 6 below.

[0698] Table 6

[0699] Figure 5 Example 6: In vivo anti-tumor experiment (1) A(％) B(％) 0 0.4 95 5 1 0.4 95 5 5 0.4 10 90 6 0.4 10 90 6.5 0.4 95 5 7 0.4 10 90 8 0.4 10 90 8.1 0.4 95 5 10 0.4 95 5

[0700] 3. Experimental results

[0701] The results are shown in Figure 6 .

[0702] The results show that under the same experimental conditions, KA0012 (Mc-AAA-DXd) and KA0010 (Mc-GGFG-DXd) can be effectively cleaved by CTSB, and can effectively release the payload, and the efficiency of the two is the same.

[0703] Figure 7

[0704] HCC827 cells (purchased from Guangzhou Jiyieuo Biological Technology Co., Ltd.) were inoculated subcutaneously on the right side of the abdomen of 5-7 week old CB-17 SCID mice (purchased from Guangdong Nanmo Biological Technology Co., Ltd.), and on the 24th day after inoculation, when the average tumor volume reached 140mm 3 left, the mice were grouped and dosed according to the tumor volume, and the modeling and specific dosing methods are shown in Table 8. After dosing, the length and width of the tumors in each group were measured, and the tumor volume was calculated.

[0705] The preparation information of each reagent is shown in Table 7 below.

[0706] Table 7

[0707]

[0708] Table 8: Dosing regimen for treating HCC827 xenograft CB-17 SCID mouse model

[0709]

[0710] The results are shown in Example 7: In vivo anti-tumor experiment (2)As shown in the figure. Compared with the isotype control, both the KA0013 monotherapy group and the KA0013+Patritumab combination group effectively inhibited tumor growth in mice. When the ratio of KA0013 to its parent monoclonal antibody Patritumab in the combination group was 1:0.5 (KA0013 1.5 mg / kg + Patritumab 0.75 mg / kg), its antitumor efficacy was stronger than that of the KA0013 monotherapy group (KA0013 1.5 mg / kg) and also better than that of the Patritumab monotherapy group (Patritumab 3 mg / kg).

[0711] like Figures 8 to 10 As shown, when the ratio of KA0013 to its parent monoclonal antibody Patritumab in the drug combination was 1:0.5 (KA0013 1.5 mg / kg + Patritumab 0.75 mg / kg), the tolerability of tumor-bearing mice was better than that of the ADC monotherapy group, indicating that the in vivo safety of the drug combination of KA0013 and anti-HER3 antibody was better than that of its KA0013 monotherapy group (KA0013 1.5 mg / kg).

[0712] Figure 8

[0713] SW620 cells (purchased from the Chinese Academy of Sciences) were subcutaneously injected into the right ventral abdomen of 4-6 week old CB-17SCID mice (purchased from Guangdong Nanmo Biotechnology Co., Ltd.). On day 22 after inoculation, when the average tumor volume reached 400 mmHg... 3 When the tumor volume was measured, the tumors were grouped according to their size and administration method. The modeling process and specific administration methods are shown in Table 10. After administration, the length and width of the tumors in each group were measured, and the tumor volume was calculated.

[0714] The preparation information for each drug is shown in Table 9 below.

[0715] Table 9

[0716]

[0717] Table 10: Dosing regimens for treating SW620 xenograft CB-17SCID mouse models

[0718]

[0719] The results are as follows Figure 9 As shown.

[0720] like Figure 10 As shown, compared with the isotype control, both the KA0013 monotherapy group and the KA0013+Patritumab combination group effectively inhibited tumor growth in mice.

[0721] like Example 8: In vivo anti-tumor experiment (3) of pharmaceutical compositionAs shown, when the ratio of KA0013 to its parent monoclonal antibody Patritumab in the drug combination was 1:0.5 (KA0013 6 mg / kg + Patritumab 3 mg / kg), the tolerability of tumor-bearing mice was better than that of the ADC monotherapy group, indicating that the in vivo safety of the drug combination of KA0013 and anti-HER3 antibody was better than that of its KA0013 monotherapy group (KA0013 6 mg / kg).

[0722] like Figures 11 to 12 As shown, the ADC concentration in the blood of mice in the KA0013+Patritumab combination group (KA0013 6 mg / kg + Patritumab 3 mg / kg) was higher than that in the KA0013 monotherapy group (KA0013 6 mg / kg).

[0723] Figure 11

[0724] SW620 cells (purchased from the Chinese Academy of Sciences) were subcutaneously injected into the right ventral abdomen of 6-7 week old CB-17SCID mice (purchased from Guangdong Nanmo Biotechnology Co., Ltd.). On day 13 after inoculation, when the average tumor volume reached 150 mm, the tumor cells were inoculated. 3 When the tumor volume was measured, the tumors were grouped according to their size and administration method. The modeling process and specific administration methods are shown in Table 11. After administration, the length and width of the tumors in each group were measured, and the tumor volume was calculated.

[0725] Table 11: Dosing regimens for treating SW620 xenograft CB-17SCID mouse models

[0726]

[0727] The results are as follows Figure 12 As shown.

[0728] like Example 9: Measurement of the bystander killing activity of Anti-HER3 antibody-drug conjugate As shown, compared with the isotype control, both the KA0013 monotherapy group and the KA0013+Patritumab combination group effectively inhibited tumor growth in mice, and the KA0013+Patritumab combination group showed stronger inhibitory activity than KA0013 monotherapy.

[0729] like Figure 13 As shown, tumor-bearing mice showed good tolerance to all groups of drugs.

[0730] Example 10: In vivo experiment (4) of pharmaceutical composition

[0731] 1. Experimental drugs

[0732] KA0013

[0733] KA0011

[0734] IgG-KA0012

[0735] IgG-KA0010

[0736] 2. Experimental method

[0737] HCC1569 cells (purchased from Nanjing Kebai Biotechnology Co., Ltd., item number: CBP60372) were routinely collected, and HCC1569 cell suspension (about 1 x 10 3 cells per well) was inoculated in a 96-well plate (Corning, item number: 7007) at 100 μL per well, and was placed in a 37°C, 5% CO2 incubator for 4 days; after 4 days, Raji-luc cells (constructed by Kangfang Biotechnology) were routinely collected, and Raji-luc cell suspension was added to the 96-well plate containing the HCC1569 cell suspension at 50 μL per well, 50 μL of gradient-diluted experimental drug (6.67, 4.44, 2.96, 1.48, 0.49, 0.049, 0.0049 nM) was added, and a blank control was designed, with 2 replicate wells per group. It was placed in a 37°C, 5% CO2 incubator for incubation for 6 days; after the incubation was completed, Luciferase (Novozyme, item number: DD1208-03) was added, and after incubation at room temperature for 10 minutes in the dark, 100 μL of supernatant was taken to a black 96-well plate (Corning, item number: 3916) after resuspension and mixing, and a multi-label microplate detector was used to detect the fluorescence value.

[0738] 3. Experimental results

[0739] The results are shown in Table 12 and Figure 14 .

[0740] Table 12: Side-killing activity of anti-HER3 antibody-drug conjugates

[0741]

[0742] The results show that the anti-HER3 antibody-drug conjugates KA0013 and KA0011 both have side-killing effects, and the side-killing activity of KA0013 is stronger than that of KA0011.

[0743] ​

[0744] Experimental drug:

[0745] Anti-Trop2 is hRS7 (G1TM) prepared in Preparation Example 5;

[0746] Anti-Trop2-MMAE is prepared in Preparation Example 6.

[0747] The experiment was carried out using 4-6 week-old hTROP2 mice (purchased from Shanghai South Model Organism Technology Co., Ltd.), and the day of administration was recorded as D0. The mice were grouped according to their body weight on the day of administration, and the specific grouping and administration methods are shown in Table 13. After administration, the skin damage of the mice was observed, and the lesion area was calculated using ImageJ software.

[0748] Table 13: Drug administration scheme of the pharmaceutical composition on hTROP2 mice

[0749]

[0750] The results are shown in ​ , wherein the D11 skin damage quantitative data show that, compared with the ISO-MMAE control group, the Anti-Trop2-MMAE group and the Anti-Trop2-MMAE + Anti-Trop2 combination group both showed obvious skin damage, but the Anti-Trop2-MMAE + Anti-Trop2 combination group significantly reduced the skin damage of the mice compared with the Anti-Trop2-MMAE group.

[0751] Although the specific embodiments of the present application have been described in detail, those skilled in the art will understand that various modifications and substitutions can be made to those details in accordance with all the teachings disclosed herein, and such changes are within the scope of protection of the present application. The entire scope of the present application is given by the appended claims and any equivalents thereof.

Claims

1. A pharmaceutical combination comprising a first active ingredient and a second active ingredient, wherein: the first active ingredient is an antibody or an antigen-binding fragment thereof; the second active ingredient is an antibody conjugate drug comprising the antibody or the antigen-binding fragment thereof, a linker, and a payload.

2. The pharmaceutical combination according to claim 1, characterized in that any one or more of the following (1) to (4): (1) the antibody is a mono-specific antibody, a bi-specific antibody, or a multi-specific antibody (e.g., a tri-specific antibody); (2) the antibody is a human IgG antibody, preferably a human IgG1 antibody or a human IgG4 antibody; (3) the antibody is an antibody for treating a tumor or an autoimmune disease; (4) the antibody is an antibody targeting a tumor-associated antigen and / or an immune checkpoint, or an antibody targeting an autoimmune disease-associated target; preferably, the tumor-associated antigen is one or more selected from the group consisting of PD-L1, CD19, CD20, CD22, CD30, CD33, CD79b, EGFR, Claudin 18.2, BCMA, HER2, HER3, TROP2, ROR1, CEACAM5, MET, tissue factor (TF), folate receptor alpha (FRa), NaPi2b, and Nectin-4; preferably, the immune checkpoint is one or more selected from the group consisting of PD-1, PD-L1, CTLA-4, LAG3, TIGIT, LMTK3, IDO, and TIM-3; preferably, the autoimmune disease-associated target is one or more selected from the group consisting of TNF-a, IL-1R, IL-2R, IL-6R, IL-7R, IL-17AR, IL-12R, IL-23R, and Integrin.

3. The pharmaceutical combination of any one of claims 1 to 2, wherein, the antibody is an anti-HER3 antibody.

4. The pharmaceutical combination of claim 3, wherein, the antibody comprises a heavy chain variable region and a light chain variable region, wherein, the heavy chain variable region comprises HCDR1 as set forth in SEQ ID NO: 1, HCDR2 as set forth in SEQ ID NO: 2, and HCDR3 as set forth in SEQ ID NO: 3; the light chain variable region comprises LCDR1 as set forth in SEQ ID NO: 4, LCDR2 as set forth in SEQ ID NO: 5, and LCDR3 as set forth in SEQ ID NO:

6.

5. The pharmaceutical combination of claim 4, wherein, the amino acid sequence of the heavy chain variable region is as set forth in SEQ ID NO: 7, and the amino acid sequence of the light chain variable region is as set forth in SEQ ID NO:

9.

6. The pharmaceutical combination according to any one of claims 1 to 5, characterized in that any one or more of the following (1) to (3): (1) the heavy chain constant region of the antibody is Ig gamma-1 chain C region or Ig gamma-4 chain C region; and the light chain constant region of the antibody is Ig kappa chain C region; (2) the amino acid sequence of the heavy chain constant region of the antibody is as set forth in SEQ ID NO: 11; and the amino acid sequence of the light chain constant region of the antibody is as set forth in SEQ ID NO: 13; (3) the antibody is a human IgGl antibody, and its heavy chain constant region has the following mutations according to the EU numbering system: L234A and L235A; L234A and G237A; L235A and G237A; or L234A, L235A and G237A.

7. The pharmaceutical combination of any one of claims 1 to 6, wherein, the antigen-binding fragment is a VHH, a scFv, a Fv fragment, a Fab fragment or a F(ab')2 fragment.

8. The pharmaceutical combination according to any one of claims 1 to 7, wherein, the linker-payload is a compound of Formula 0 or a pharmaceutically acceptable salt or ester thereof; the compound of Formula 0 comprises a compound of Formula III, n alanines, and a compound of Formula IV connected in that order, wherein, n is 2, 3, 4, 5 or 6; the compound of Formula III, the n alanines, and the compound of Formula IV are independently connected to each other two by two either directly or through a chemical group.

9. The pharmaceutical combination of claim 8, wherein, the compound of Formula 0 is shown in the following Formula I, wherein, A represents an alanine; m is 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12; n is 2, 3, 4, 5 or 6.

10. The pharmaceutical combination according to any one of claims 8 to 9, wherein, the compound of Formula 0 is shown in the following Formula II, 11. The pharmaceutical combination of any one of claims 1 to 7, wherein, the linker is a hydrazone bond, a disulfide bond, a thioether bond or a peptide bond, or a chemical group comprising a hydrazone bond, a disulfide bond, a thioether bond or a peptide bond; preferably, the linker is one or more selected from 6-maleimidocaproyl (MC), maleimidopropionyl (MP), valine-citrulline (Val-Cit), alanine-phenylalanine (Ala-Phe), alanine-alanine-alanine (Ala-Ala-Ala), Mc-Ala-Ala-Ala, p-aminobenzyloxycarbonyl (PAB), 6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (MC-Val-Cit-PAB), Mal-PEGn-Val-Cit-PAB (n is a positive integer selected from 1-20), Phe-Lys(Fmoc)-PAB, Aloc-D-Ala-Phe-Lys(Aloc)-PAB-PNP, Boc-Phe-(Alloc)Lys-PAB-PNP and 3-(pyridine-2-yl disulfide) propionic acid perfluorophenyl ester.

12. The pharmaceutical combination of any one of claims 1 to 7 and 11, wherein, the payload is one or more selected from a tubulin inhibitor, a DNA damaging agent, a topoisomerase inhibitor, an ALK inhibitor and a PARP inhibitor; Preferably, the tubulin inhibitor is one or more selected from the group consisting of dolastatins, auristatin cytotoxic molecules, and maytansine cytotoxic molecules; preferably, the auristatin cytotoxic molecule is selected from the group consisting of monomethyl auristatin E (MMAE) and derivatives thereof, and monomethyl auristatin F (MMAF) and derivatives thereof; preferably, the maytansine cytotoxic molecule is selected from the group consisting of DM1 and derivatives thereof, and DM4 and derivatives thereof; Preferably, the DNA damaging agent is one or more selected from the group consisting of calicheamicin, duocarmycin, and antracycline derivatives (PBDs); Preferably, the topoisomerase inhibitor is selected from the group consisting of camptothecins and camptothecin derivatives; preferably, the camptothecin and camptothecin derivative is 7-ethyl-10-hydroxy camptothecin (SN-38) or DXd; Preferably, the PARP inhibitor is Niraparib.

13. The pharmaceutical combination of any one of claims 1 to 12, wherein the linker is linked to the antibody or antigen-binding fragment thereof via one or more thioether bonds. Preferably, the linker is linked to the sulfur atom of the disulfide bond position of the antibody hinge region via a thioether bond.

14. The pharmaceutical combination according to any one of claims 1 to 13, wherein, The average number of linker-payload conjugated to each antibody molecule is 1-12; Preferably, the average number of linker-payload conjugated to each antibody molecule is 1-8.

15. The pharmaceutical combination of any one of claims 1 to 14, wherein, the mass ratio or molar ratio of the first active ingredient to the second active ingredient is (0.1-30): 1, preferably (0.3-20): 1, (0.4-15): 1, (0.5-15): 1, (0.5-10): 1, (0.5-5): 1, (0.8-5): 1, (0.8-3): 1, (1-3): 1, (1.2-3): 1, (1.5-2.5): 1, (1.8-2.2): 1, 0.1: 1, 0.2: 1, 0.3: 1, 0.4: 1, 0.5: 1, 0.6: 1, 0.7: 1, 0.8: 1, 0.9: 1, 1: 1, 1.1: 1, 1.2: 1, 1.3: 1, 1.4: 1, 1.5: 1, 1.6: 1, 1.7: 1, 1.8: 1, 1.9: 1, 2: 1, 2.1: 1, 2.2: 1, 2.3: 1, 2.4: 1, or 2.5:

1.

16. The pharmaceutical combination according to any one of claims 1 to 15, wherein, the first active ingredient and the second active ingredient are mixed together, and the pharmaceutical combination is a pharmaceutical composition; or the first active ingredient and the second active ingredient are in separate packages from each other, and the pharmaceutical combination is a combination product. Preferably, the pharmaceutical combination further comprises one or more pharmaceutically acceptable excipients.

17. The pharmaceutical combination of claim 1, which is selected from (1) to (3) below: (1) The pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein: the first active ingredient is an anti-HER3 antibody or an antigen-binding fragment thereof, the second active ingredient is an antibody conjugate drug; wherein the antibody conjugate drug comprises the anti-HER3 antibody or the antigen-binding fragment thereof, a linker, and a payload, wherein the linker-payload is a compound of Formula 0 or a pharmaceutically acceptable salt or ester thereof; wherein the compound of Formula 0 is shown below as Formula II, wherein the anti-HER3 antibody comprises a heavy chain variable region and a light chain variable region, wherein: the heavy chain variable region comprises HCDR1 shown in SEQ ID NO: 1, HCDR2 shown in SEQ ID NO: 2, and HCDR3 shown in SEQ ID NO: 3; the light chain variable region comprises LCDR1 shown in SEQ ID NO: 4, LCDR2 shown in SEQ ID NO: 5, and LCDR3 shown in SEQ ID NO: 6; wherein the average number of linker-payload conjugated to each anti-HER3 antibody molecule is 1-8; Preferably, the mass ratio or molar ratio of the first active ingredient to the second active ingredient is (1:2) to (2:1). (2) The pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein: the first active ingredient is an anti-HER3 antibody or an antigen-binding fragment thereof, the second active ingredient is an antibody conjugate drug; wherein the antibody conjugate drug comprises the anti-HER3 antibody or the antigen-binding fragment thereof, a linker, and a payload, wherein the linker-payload is a compound of Formula 0 or a pharmaceutically acceptable salt or ester thereof; wherein the compound of Formula 0 is shown below as Formula II, wherein the anti-HER3 antibody comprises a heavy chain variable region and a light chain variable region, wherein: the amino acid sequence of the heavy chain variable region of the anti-HER3 antibody is shown in SEQ ID NO: 7, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 9; wherein the average number of linker-payload conjugated to each anti-HER3 antibody molecule is 1-8; Preferably, the mass ratio or molar ratio of the first active ingredient to the second active ingredient is (1:2) to (2:1). (3) The pharmaceutical combination comprises a first active ingredient and a second active ingredient, wherein: the first active ingredient is an anti-HER3 antibody or an antigen-binding fragment thereof, the second active ingredient is an antibody conjugate drug; wherein the antibody conjugate drug comprises the anti-HER3 antibody or the antigen-binding fragment thereof, a linker, and a payload, wherein the linker-payload is a compound of Formula 0 or a pharmaceutically acceptable salt or ester thereof; wherein the compound of Formula 0 is shown below as Formula II, wherein the anti-HER3 antibody comprises a heavy chain variable region and a light chain variable region, wherein: the amino acid sequence of the heavy chain variable region of the anti-HER3 antibody is shown in SEQ ID NO: 7, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 9; the amino acid sequence of the heavy chain variable region of the anti-HER3 antibody is set forth in SEQ ID NO: 7 and the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO: 11, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 9 and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO: 13; wherein the average number of linkers-payloads coupled to each anti-HER3 antibody molecule is 1-8; Preferably, the mass ratio or molar ratio of the first active ingredient to the second active ingredient is (1:2) to (2:1).

18. The pharmaceutical combination according to any one of claims 1 to 17 for use in the treatment or prevention of a tumor or an autoimmune disease; Preferably, the tumor is a HER3-positive tumor; Preferably, the tumor is selected from one or more of lung cancer, colon cancer, rectal cancer, breast cancer, clear cell sarcoma, skin cancer, renal cancer, urothelial cancer, prostate cancer, glioblastoma multiforme, ovarian cancer, pancreatic cancer, melanoma, liver cancer, bladder cancer, gastric cancer, gastrointestinal stromal tumor, cervical cancer, head and neck cancer, esophageal cancer, epidermoid cancer, peritoneal cancer, adult glioblastoma multiforme, uterine cancer, salivary gland cancer, vulvar cancer, thyroid cancer, anal cancer, and penile cancer; Preferably, the lung cancer is non-small cell lung cancer, e.g., lung adenocarcinoma; Preferably, the colon cancer is metastatic colon cancer; Preferably, the autoimmune disease is selected from one or more of systemic lupus erythematosus, rheumatoid arthritis, Sjogren's syndrome, ankylosing spondylitis, multiple sclerosis, autoimmune thyroid disease, and inflammatory bowel disease.

19. Use of the pharmaceutical combination according to any one of claims 1 to 17 for the manufacture of a medicament for the treatment or prevention of a tumor or an autoimmune disease; Preferably, the tumor is a HER3-positive tumor; Preferably, the tumor is selected from one or more of lung cancer, colon cancer, rectal cancer, breast cancer, clear cell sarcoma, skin cancer, renal cancer, urothelial cancer, prostate cancer, glioblastoma multiforme, ovarian cancer, pancreatic cancer, melanoma, liver cancer, bladder cancer, gastric cancer, gastrointestinal stromal tumor, cervical cancer, head and neck cancer, esophageal cancer, epidermoid cancer, peritoneal cancer, adult glioblastoma multiforme, uterine cancer, salivary gland cancer, vulvar cancer, thyroid cancer, anal cancer, and penile cancer; Preferably, the lung cancer is non-small cell lung cancer, e.g., lung adenocarcinoma; Preferably, the colon cancer is metastatic colon cancer; Preferably, the autoimmune disease is selected from one or more of systemic lupus erythematosus, rheumatoid arthritis, Sjogren's syndrome, ankylosing spondylitis, multiple sclerosis, autoimmune thyroid disease, and inflammatory bowel disease.

Images