Use of bispecific antibody-drug conjugate in treating cancer

Abstract

Provided is use of an anti-EGFR / c-Met antibody-drug conjugate and a tautomer thereof, a mesomer thereof, a racemate thereof, an enantiomer thereof, a diastereomer thereof, a pharmaceutically acceptable salt thereof, a hydrate thereof, a solvate thereof, or an isotopically labeled compound thereof in the preparation of a drug for preventing and / or treating cancer.

Description

Uses of bispecific antibody-drug conjugates in cancer treatment Technical Field

[0001] This application belongs to the pharmaceutical field and relates to the use of a bispecific antibody-drug conjugate in the preparation of drugs for the prevention and / or treatment of cancer. Specifically, the present invention provides the use of a bispecific antibody-drug conjugate and its tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates, or isotope-labeled derivatives in the preparation of drugs for the prevention and / or treatment of cancer. Background Technology

[0002] Lung cancer is one of the most common and deadliest malignant tumors in the world. Data from 2022 shows that lung cancer is the leading cause of cancer morbidity and mortality in my country, accounting for over 20% of new cases. Non-small cell lung cancer (NSCLC) accounts for 80%–85% of all lung cancers, and approximately 70% of these cases are diagnosed at a locally advanced or metastatic stage, making surgical resection impossible. Furthermore, most patients who undergo early surgical resection experience distant metastasis upon recurrence, often leading to death.

[0003] EGFR mutations are one of the major driver genes in NSCLC, accounting for 10%–17% in Western populations and 30%–50% in Asian populations. EGFR mutations typically occur in exons 18–21, with exon 19 deletion (Ex19del) and exon 21 L858R mutations being the most common types, accounting for over 85% of all EGFR mutations. These patients respond well to EGFR tyrosine kinase inhibitors (TKIs). First-generation EGFR TKIs gefitinib and erlotinib, second-generation EGFR TKIs afatinib and dacomitinib, and third-generation EGFR TKI osimertinib are currently the standard first-line treatments for NSCLC patients with EGFR-sensitive mutations. However, after a period of treatment (approximately 10–18 months), drug resistance and disease progression inevitably occur in all these cases.

[0004] Recent preclinical and clinical studies have reported several EGFR-dependent and EGFR-independent resistance mechanisms. Among these, MET gene amplification and protein overexpression are important resistance mechanisms to EGFR-TKI targeted therapy, with secondary MET gene amplification occurring in 5%–50% of EGFR-TKI resistant patients. Furthermore, drugs targeting EGFR and c-MET have shown clinical benefit in EGFR-mutant NSCLC. Treatment of NSCLC after EGFR TKI resistance has become a clinical hot topic, as these patients require better treatment options to achieve survival benefits. Developing combination therapy strategies targeting MET amplification or overexpression resistance mechanisms may lead to better clinical outcomes for these patients.

[0005] Therefore, there is an urgent need to develop new drugs that target EGFR / c-MET to achieve better therapeutic effects in multiple tumor types such as NSCLC and HNSCC with high expression of EGFR and / or c-MET, thereby maximizing patient survival benefits. Summary of the Invention

[0006] This disclosure provides methods for treating cancer and pharmaceutical uses of anti-EGFR / c-Met antibody-drug conjugates and their tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates or isotope-labeled compounds.

[0007] In some implementations, this disclosure provides for any of the following uses:

[0008] (1) Use of anti-EGFR / c-Met antibody-drug conjugates and their tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates or isotope labels in the preparation of drugs for treating cancer.

[0009] (2) Anti-EGFR / c-Met antibody-drug conjugates and their tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates or isotope labels for the prevention and / or treatment of cancer.

[0010] (3) Anti-EGFR / c-Met antibody-drug conjugates and their tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates or isotopic labels are used for the prevention and / or treatment of subjects with cancer, wherein the anti-EGFR / c-Met antibody-drug conjugate is administered to the subject.

[0011] This disclosure also provides a method for treating cancer, the method comprising administering to a subject suffering from cancer a therapeutically effective amount of an anti-EGFR / c-Met antibody-drug conjugate and its tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates or isotopic labels.

[0012] This disclosure also provides a method for treating a subject with cancer, the method comprising administering to the subject a therapeutically effective amount of an anti-EGFR / c-Met antibody-drug conjugate and its tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates, or isotope-labeled compounds.

[0013] This disclosure also provides a method for inhibiting the growth or proliferation of cells expressing EGFR and / or c-Met, the method comprising contacting the cells with an anti-EGFR / c-Met antibody-drug conjugate and its tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates or isotope-labeled substances.

[0014] This disclosure also provides a method for inhibiting the growth or metastasis of tumor or cancer cells expressing EGFR and / or c-Met in a subject, the method comprising administering to the subject a therapeutically effective amount of an anti-EGFR / c-Met antibody-drug conjugate and its tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates, or isotopic markers to inhibit the growth or metastasis of tumor or cancer cells expressing EGFR and / or c-Met.

[0015] In some implementations, this disclosure provides a product as shown in any of the following:

[0016] (1) A pharmaceutical composition comprising an anti-EGFR / c-Met antibody-drug conjugate and its tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates or isotopes;

[0017] (2) A kit containing an anti-EGFR / c-Met antibody-drug conjugate and its tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates or isotopes.

[0018] (3) Products, including anti-EGFR / c-Met antibody-drug conjugates and their tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates or isotopes.

[0019] In some implementations, this disclosure provides for any of the following uses:

[0020] (1) Use of anti-EGFR / c-Met antibody drug conjugates and their tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates or isotopes in the preparation of medicaments for the treatment of cancer.

[0021] (2) Pharmaceutical compositions, kits or articles including anti-EGFR / c-Met antibody drug conjugates and their tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates or isotopes for the prevention and / or treatment of cancer.

[0022] (3) A pharmaceutical composition, kit or article comprising an anti-EGFR / c-Met antibody drug conjugate and its tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates or isotopes for the prevention and / or treatment of a subject suffering from cancer, wherein the pharmaceutical composition, kit or article is administered to the subject.

[0023] In some implementations, the structures of the above-mentioned anti-EGFR / c-Met antibody drug conjugates and their tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates, or isotope-labeled compounds are shown in general formula (1).

[0024] Wherein: BsAb is an anti-EGFR / c-Met antibody or its antigen-binding fragment, comprising: at least one antigen-binding module 1 specifically binding to EGFR and at least one antigen-binding module 2 specifically binding to c-Met; the antigen-binding module 1 comprises a heavy chain variable region 1 (VH1) and a light chain variable region 1 (VL1), and the antigen-binding module 2 comprises a heavy chain variable region 2 (VH2) and a light chain variable region 2 (VL2), wherein i) the VH1 comprises the heavy chain complementarity-determining region (HCDR)1 (HCDR1), HCDR2, and HCDR3 amino acid sequences of SEQ ID NO:07, 08, and 09, respectively; and ii) the VL1 comprises the light chain complementarity-determining region (LCDR)1 (LCDR1), LCDR2, and LCDR3 amino acid sequences of SEQ ID NO:10, 11, and 12, respectively; iii) the VH2 comprises the HCDR1, HCDR2, and HCDR3 amino acid sequences of SEQ ID NO:01, 02, and 03, respectively; and iv) the VL2 comprises the heavy chain complementarity-determining region (LCDR)1 (LCDR1), LCDR2, and LCDR3 amino acid sequences of SEQ ID NO:10, 11, and 12, respectively; The amino acid sequences of LCDR1, LCDR2, and LCDR3 in NO:04, 05, and 06;

[0025] y is selected from decimals or integers from 1 to 12;

[0026] -L- represents a connector that covalently links BsAb to D;

[0027] D is a cytotoxic drug, an immunostimulant, or a radioactive isotope, preferably a DNA topoisomerase inhibitor, a tubulin inhibitor, a DNA damaging agent, antimetabolite, or an antitumor antibiotic; more preferably, camptothecin or its analogues or derivatives, erribulin or its analogues or derivatives, auristatin derivatives, maytansine or its analogues or derivatives, dolastatin or its derivatives, calicacin or pyrrolobenzodiazepine (PBD); further preferably, exatecan or its analogues or derivatives, erribulin or its analogues or derivatives, MMAE, MMAF, MMAD, SN-38, DM1, or DM4.

[0028] In some embodiments, the above-mentioned anti-EGFR / c-Met antibody or its antigen-binding fragment: i) the above-mentioned VH1 contains an amino acid sequence having at least 90%, 95% or 99% sequence identity with SEQ ID NO: 15, and the above-mentioned VL1 contains an amino acid sequence having at least 90%, 95% or 99% sequence identity with SEQ ID NO: 16; ii) the above-mentioned VH2 contains an amino acid sequence having at least 90%, 95% or 99% sequence identity with SEQ ID NO: 13, and the above-mentioned VL2 contains an amino acid sequence having at least 90%, 95% or 99% sequence identity with SEQ ID NO: 14.

[0029] In a preferred embodiment, the above-mentioned anti-EGFR / c-Met antibody or its antigen-binding fragment: i) the VH1 contains the sequence shown in SEQ ID NO: 15, and the VL1 contains the sequence shown in SEQ ID NO: 16; ii) the VH2 contains the sequence shown in SEQ ID NO: 13, and the VL2 contains the sequence shown in SEQ ID NO: 14.

[0030] In a preferred embodiment, the above-mentioned anti-EGFR / c-Met antibody or its antigen-binding fragment: i) the VH1 sequence is as shown in SEQ ID NO: 15, and the VL1 sequence is as shown in SEQ ID NO: 16; ii) the VH2 sequence is as shown in SEQ ID NO: 13, and the VL2 sequence is as shown in SEQ ID NO: 14.

[0031] In some embodiments, the above-mentioned anti-EGFR / c-Met antibody or its antigen-binding fragment comprises: a) a first heavy chain (HC1) containing VH1 and an HC1 heavy chain constant region (HC1_CH), b) a second heavy chain (HC2) containing VH2 and an HC2 heavy chain constant region (HC2_CH), c) a first light chain (LC1) containing VL1 and an LC1 light chain constant region (LC1_CL), and d) a second light chain (LC2) containing VL2 and an LC2 light chain constant region (LC2_CL); wherein i) the above-mentioned HC1_CH and HC2_CH contain an IgG heavy chain constant region or a variant thereof, preferably containing an IgG1, IgG2, IgG3 or IgG4 heavy chain constant region or a variant thereof, more preferably containing a sequence as shown in SEQ ID NO: 17 or a variant thereof, ii) the above-mentioned LC1_CL and LC2_CL contain a k or λ light chain constant region or a variant thereof, preferably containing a k light chain constant region or a variant thereof; more preferably containing a sequence as shown in SEQ ID NO: 17 or a variant thereof. NO: 18 The sequence or a variant thereof; wherein, the variant described in any one of i), ii) has at least 80%, 85%, 90%, 95%, or 99% sequence identity with the sequence from which it is derived, or the variant has at most 20, 15, 10, 9, 8, 7, 6, or 5 amino acid substitutions, deletions, or additions compared with the sequence from which it is derived.

[0032] In some embodiments, the above-mentioned HC1_CH and HC2_CH contain sequences as shown in SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21 or SEQ ID NO: 22, and the above-mentioned LC1_CL and LC2_CL contain sequences as shown in SEQ ID NO: 18, SEQ ID NO: 23, SEQ ID NO: 24;

[0033] In a preferred embodiment, the HC1_CH contains a sequence as shown in SEQ ID NO: 17, SEQ ID NO: 19, or SEQ ID NO: 21; the HC2_CH contains a sequence as shown in SEQ ID NO: 17, SEQ ID NO: 20, or SEQ ID NO: 22; the LC1_CL contains a sequence as shown in SEQ ID NO: 18 or SEQ ID NO: 23; and the LC2_CL contains a sequence as shown in SEQ ID NO: 18 or SEQ ID NO: 24.

[0034] In some embodiments, HC1 contains a sequence as shown in SEQ ID NO: 25 or SEQ ID NO: 29, HC2 contains a sequence as shown in SEQ ID NO: 26 or SEQ ID NO: 30, LC1 contains a sequence as shown in SEQ ID NO: 27 or SEQ ID NO: 31, and LC2 contains a sequence as shown in SEQ ID NO: 28 or SEQ ID NO: 32.

[0035] In a preferred embodiment, HC1 comprises the sequence shown in SEQ ID NO: 25, HC2 comprises the sequence shown in SEQ ID NO: 26, LC1 comprises the sequence shown in SEQ ID NO: 27, and LC2 comprises the sequence shown in SEQ ID NO: 28.

[0036] In a preferred embodiment, HC1 comprises the sequence shown in SEQ ID NO: 29, HC2 comprises the sequence shown in SEQ ID NO: 30, LC1 comprises the sequence shown in SEQ ID NO: 31, and LC2 comprises the sequence shown in SEQ ID NO: 32.

[0037] In a preferred embodiment, the HC1 sequence is shown in SEQ ID NO: 25, the HC2 sequence is shown in SEQ ID NO: 26, the LC1 sequence is shown in SEQ ID NO: 27, and the LC2 sequence is shown in SEQ ID NO: 28.

[0038] In a preferred embodiment, the HC1 sequence is shown in SEQ ID NO: 29, the HC2 sequence is shown in SEQ ID NO: 30, the LC1 sequence is shown in SEQ ID NO: 31, and the LC2 sequence is shown in SEQ ID NO: 32.

[0039] In some embodiments, D is selected from the following compounds, and their pharmaceutically acceptable salts, solvates, tautomers, meso compounds, racemates, enantiomers, and diastereomers.

[0040] In the preferred embodiment, D is selected from the following compounds and their pharmaceutically acceptable salts, solvates, tautomers, meso compounds, racemates, enantiomers, and diastereomers.

[0041] In some embodiments, the D group is linked to the -L- group in the antibody-drug conjugate via a hydroxyl, amino, primary, secondary, or tertiary amino group thereon.

[0042] In some implementations, the structures of the above-mentioned anti-EGFR / c-Met antibody drug conjugates and their tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates, or isotope-labeled compounds are shown in general formulas (2)-(12).

[0043] Wherein, BsAb is as defined above, -L- is as defined above, and y is selected from an integer or decimal of 1-12, preferably 2-8, more preferably 2-6 or 3-8, and even more preferably 2, 3, 4, 5, 6, or 7.

[0044] In a preferred embodiment, the structures of the above-mentioned anti-EGFR / c-Met antibody drug conjugates and their tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates, or isotope-labeled compounds are shown in general formulas (11)-(12).

[0045] Wherein, BsAb is as defined above, -L- is as defined above, and y is selected from an integer or decimal of 1-12, preferably 2-8, more preferably 2-6 or 3-8, and even more preferably 2, 3, 4, 5, 6, or 7.

[0046] In some embodiments, the -L- is selected from -L1-L2-L3-L4-L5-, wherein -L1- is a covalent linking unit covalently linked to BsAb, -L2- is an extension unit, -L3- is selected from a bond or a polar hydrophilic group, -L4- is selected from peptide residues consisting of 1-8 amino acids, and -L5- is a bond or a self-cleaving fragment.

[0047] In one implementation, the above -L1- is selected from:

[0048] The above -L2- is selected from: -(CH2)s 1 -C(=O)-, -CH2-C(=O)-NH-(CH2)s 1 -C(=O)-,-(CH2CH2O)s 2 -(CH2)s 1 -C(=O)-, Among them, S 1 and S 2 Each number is independently selected from 0-8;

[0049] The above -L3- is selected from the key, Among them, S 3 Selected from 2-12, with 6-12 being the preferred choice;

[0050] The -L4- group is selected from peptide residues consisting of 1-8 amino acids, wherein the amino acids are selected from phenylalanine, isoleucine, leucine, tryptophan, valine, methionine, tyrosine, alanine, threonine, histidine, serine, glutamine, arginine, lysine, asparagine, glutamic acid, proline, citrulline, aspartic acid, and glycine; preferably, -L4- is selected from:

[0051] -L5- is selected from: bond, -NH-CH2-, or In some implementations, the L above is selected from:

[0052] In some implementations, the structures of the above-mentioned anti-EGFR / c-Met antibody drug conjugates and their tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates or isotope-labeled compounds are shown in general formulas (13)-(24).

[0053] Wherein, D is selected from the following compounds and their pharmaceutically acceptable salts, solvates, tautomers, meso compounds, racemates, enantiomers, and diastereomers:

[0054] Wherein, BsAb is as defined above, and y is selected from an integer or decimal of 1-12, preferably 2-8, more preferably 2-6 or 3-8, and even more preferably 2, 3, 4, 5, 6, or 7.

[0055] In some embodiments, the structures of the above-mentioned anti-EGFR / c-Met antibody drug conjugates and their tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates, or isotope-labeled derivatives are selected from the following:

[0056] Wherein, BsAb is as defined above, and y is selected from an integer or decimal of 1-12, preferably 2-8, more preferably 2-6 or 3-8, and even more preferably 2, 3, 4, 5, 6, or 7.

[0057] In a preferred embodiment, the structures of the above-mentioned anti-EGFR / c-Met antibody drug conjugate and its tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates, or isotope-labeled compounds are selected from the following:

[0058] Wherein, BsAb is as defined above, and y is selected from an integer or decimal of 1-12, preferably 2-8, more preferably 2-6 or 3-8, and even more preferably 2, 3, 4, 5, 6, or 7.

[0059] In some embodiments, the single-dose dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 1.0 mg / kg to 12.0 mg / kg, preferably 1.0 mg / kg, 2.0 mg / kg, 3.0 mg / kg, 4.0 mg / kg, 4.2 mg / kg, 4.5 mg / kg, 4.8 mg / kg, 5.0 mg / kg, 6.0 mg / kg, 7.0 mg / kg, 8.0 mg / kg, 9.0 mg / kg, 10.0 mg / kg, 11.0 mg / kg, and 12.0 mg / kg, more preferably 3.0 mg / kg and 4.0 mg / kg. The dosages are 4.2 mg / kg, 4.5 mg / kg, 4.8 mg / kg, 5.0 mg / kg, 6.0 mg / kg, 7.0 mg / kg, 8.0 mg / kg, 9.0 mg / kg, and 10.0 mg / kg; more preferably 3.0 mg / kg, 4.0 mg / kg, 4.2 mg / kg, 4.5 mg / kg, 4.8 mg / kg, 5.0 mg / kg, 6.0 mg / kg, 7.0 mg / kg, and 8.0 mg / kg; the dosing frequency is once a week, once every two weeks, once every three weeks, or once every four weeks, preferably once every two weeks or once every three weeks.

[0060] In a preferred embodiment, the single-dose dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 3.0 mg / kg, 4.0 mg / kg, 4.2 mg / kg, 4.5 mg / kg, 4.8 mg / kg, 5.0 mg / kg, 6.0 mg / kg, 7.0 mg / kg, 8.0 mg / kg, 9.0 mg / kg, and 10.0 mg / kg; the dosing frequency is once every two weeks and once every three weeks.

[0061] In a preferred embodiment, the single-dose dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 3.0 mg / kg, 4.0 mg / kg, 4.2 mg / kg, 4.5 mg / kg, 4.8 mg / kg, 5.0 mg / kg, 6.0 mg / kg, 7.0 mg / kg, 8.0 mg / kg, and 9.0 mg / kg; the dosing frequency is once every three weeks.

[0062] In a preferred embodiment, the single-dose dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 3.0 mg / kg, 4.0 mg / kg, 4.2 mg / kg, 4.5 mg / kg, 4.8 mg / kg, 5.0 mg / kg, 6.0 mg / kg, 7.0 mg / kg, 8.0 mg / kg, and 9.0 mg / kg; the dosing frequency is once every two weeks.

[0063] In a preferred embodiment, the single dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 3.0 mg / kg; the dosing frequency is once every three weeks. In a preferred embodiment, the single dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 4.0 mg / kg; the dosing frequency is once every three weeks. In a preferred embodiment, the single dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 4.2 mg / kg; the dosing frequency is once every three weeks. In a preferred embodiment, the single dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 4.5 mg / kg; the dosing frequency is once every three weeks. In a preferred embodiment, the single dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 4.8 mg / kg; the dosing frequency is once every three weeks. In a preferred embodiment, the single dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 5.0 mg / kg; the dosing frequency is once every three weeks. In a preferred embodiment, the single-dose dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 6.0 mg / kg; the dosing frequency is once every three weeks. In a preferred embodiment, the single-dose dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 7.0 mg / kg; the dosing frequency is once every three weeks. In a preferred embodiment, the single-dose dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 8.0 mg / kg; the dosing frequency is once every three weeks. In a preferred embodiment, the single-dose dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 9.0 mg / kg; the dosing frequency is once every three weeks.

[0064] In a preferred embodiment, the single dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 3.0 mg / kg, and the dosing frequency is once every two weeks. In a preferred embodiment, the single dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 4.0 mg / kg, and the dosing frequency is once every two weeks. In a preferred embodiment, the single dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 4.2 mg / kg, and the dosing frequency is once every two weeks. In a preferred embodiment, the single dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 4.5 mg / kg, and the dosing frequency is once every two weeks. In a preferred embodiment, the single dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 4.8 mg / kg, and the dosing frequency is once every two weeks. In a preferred embodiment, the single dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 5.0 mg / kg, and the dosing frequency is once every two weeks. In a preferred embodiment, the single-dose dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 6.0 mg / kg; the dosing frequency is once every two weeks. In a preferred embodiment, the single-dose dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 7.0 mg / kg; the dosing frequency is once every two weeks. In a preferred embodiment, the single-dose dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 8.0 mg / kg; the dosing frequency is once every two weeks. In a preferred embodiment, the single-dose dose of the above-mentioned anti-EGFR / c-Met antibody-drug conjugate is 9.0 mg / kg; the dosing frequency is once every two weeks.

[0065] In some implementations, the aforementioned anti-EGFR / c-Met antibody-drug conjugate is administered intravenously.

[0066] In some implementations, the aforementioned anti-EGFR / c-Met antibody-drug conjugate is administered subcutaneously.

[0067] In a preferred embodiment, the aforementioned cancers are lung cancer, head and neck cancer, colorectal cancer, nasopharyngeal cancer, esophageal cancer, esophagogastric junction cancer, oral cancer, oropharyngeal cancer, hypopharyngeal cancer, laryngeal cancer, tongue cancer, stomach cancer, liver cancer, kidney cancer, pancreatic cancer, spleen cancer, skin cancer, thymic cancer, thyroid cancer, prostate cancer, bladder cancer, vaginal cancer, testicular cancer, anal cancer, breast cancer, ovarian cancer, cervical cancer, epithelial cell carcinoma, or glioblastoma.

[0068] In a preferred embodiment, the aforementioned cancers are non-small cell lung cancer (NSCLC), small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, large cell lung cancer, head and neck squamous cell carcinoma, head and neck adenocarcinoma, colorectal adenocarcinoma, colorectal mucinous carcinoma, oral cavity, oropharynx, hypopharynx or larynx squamous cell carcinoma, nasopharyngeal carcinoma, esophageal adenocarcinoma, esophageal squamous cell carcinoma, esophagogastric junction adenocarcinoma, and esophagogastric junction squamous cell carcinoma.

[0069] In a preferred embodiment, the aforementioned cancers are non-squamous NSCLC, squamous NSCLC, head and neck squamous cell carcinoma, colorectal adenocarcinoma, oral cavity, oropharynx, hypopharynx or larynx squamous cell carcinoma, nasopharyngeal carcinoma, esophageal squamous cell carcinoma, and esophagogastric junction squamous cell carcinoma.

[0070] In some implementations, the aforementioned cancers are EGFR and / or c-Met cancers that are associated with wild-type EGFR, EGFR mutations, EGFR gene amplification, elevated levels of circulating HGF, wild-type c-Met, c-Met mutations, c-Met gene amplification, or mutant KRAS.

[0071] In some implementations, the aforementioned EGFR mutation includes at least one mutation selected from the group consisting of: L718Q, G719A, G719X (X being any amino acid), E746K, L747S, E749Q, A750P, A755V, V765M, T790M, C797S, L858R, L858P, L861X (X being any amino acid) substitutions; E746-A750 deletions; R748-P753 deletions; Ala(A) insertions between M766 and A767; Ser, Val, and Ala(SVA) insertions between S768 and V769; and Asn and Ser(NS) insertions between P772 and H773. Insertion of one or more amino acids between D761 and E762, between A763 and Y764, between Y764 and Y765, between M766 and A767, between A767 and V768, between S768 and V769, between V769 and D770, between D770 and N771, between N771 and P772, between P772 and H773, between H773 and V774, and between V774 and C775; deletion of one or more of EGFR exon 19 and EGFR exon 20; insertion of one or more of EGFR exon 19, EGFR exon 20, S768I, L861Q, and G719X (where X is any amino acid).

[0072] In a preferred embodiment, the above-mentioned EGFR mutation includes: exon 19 deletion (Ex19del), L858R mutation, or a combination thereof.

[0073] In some implementations, the aforementioned cancer refers to cancer that has failed, progressed, or is intolerable to one or more previous anticancer therapies.

[0074] In some implementations, the aforementioned one or more anticancer therapies include one or more chemotherapeutic agents, checkpoint inhibitors, targeted anticancer therapies, or kinase inhibitors, or any combination thereof.

[0075] In a preferred embodiment, one or more of the above-mentioned anticancer therapies include carboplatin, cisplatin, paclitaxel, gemcitabine, vinorelbine, docetaxel, PD-(L)1 inhibitors, EGFR inhibitors, VEGFR inhibitors, c-Met inhibitors, HER2 inhibitors, HER3 inhibitors, HER4 inhibitors, or any combination thereof.

[0076] In some implementation schemes, the aforementioned cancers are defined as failure of adequate standard treatment or intolerance to standard treatment.

[0077] In some implementations, the standard treatment described above includes one or more chemotherapeutic agents, checkpoint inhibitors, targeted anticancer therapies, or kinase inhibitors, or any combination thereof.

[0078] In a preferred embodiment, the above standard treatment includes carboplatin, cisplatin, paclitaxel, gemcitabine, vinorelbine, docetaxel, PD-1 inhibitors, PD-L1 inhibitors, EGFR inhibitors, VEGFR inhibitors, c-Met inhibitors, HER2 inhibitors, HER3 inhibitors, HER4 inhibitors, or any combination thereof.

[0079] In some implementation schemes, the aforementioned cancers are locally advanced or metastatic NSCLC, recurrent or metastatic head and neck squamous cell carcinoma, recurrent or metastatic colorectal adenocarcinoma, recurrent or metastatic oral, oropharyngeal, hypopharyngeal or laryngeal squamous cell carcinoma, metastatic nasopharyngeal carcinoma, recurrent or metastatic esophageal squamous cell carcinoma, and recurrent or metastatic esophagogastric junction squamous cell carcinoma.

[0080] In a preferred embodiment, the aforementioned cancers are locally advanced or metastatic NSCLC carrying EGFR-sensitive mutations, metastatic NSCLC with EGFR wild-type, recurrent or metastatic head and neck squamous cell carcinoma that has failed or is intolerant to standard treatment, recurrent or metastatic colorectal adenocarcinoma that has failed or is intolerant to standard treatment, recurrent or metastatic squamous cell carcinoma of the oral cavity, oropharynx, hypopharynx, or larynx that has failed or is intolerant to standard treatment, metastatic nasopharyngeal carcinoma that has failed or is intolerant to adequate standard treatment, recurrent or metastatic esophageal squamous cell carcinoma that has failed or is intolerant to standard treatment, and recurrent or metastatic esophagogastric junction squamous cell carcinoma that has failed or is intolerant to standard treatment.

[0081] In a preferred embodiment, the aforementioned cancers are: locally advanced or metastatic NSCLC with EGFR-sensitive mutations that have undergone systemic treatment with EGFR TKIs; metastatic NSCLC with EGFR wild-type cells that have previously received platinum-based chemotherapy or PD-1 / L1 inhibitor monotherapy or combination therapy; recurrent or metastatic head and neck squamous cell carcinoma that has previously received platinum-based chemotherapy or PD-1 / L1 inhibitor monotherapy or combination therapy; recurrent or metastatic colorectal adenocarcinoma that has failed or is intolerant to standard treatment and does not carry KRAS mutations, NRAS mutations, or BRAF V600E mutations; recurrent or metastatic squamous cell carcinoma of the oral cavity, oropharynx, hypopharynx, or larynx that has previously received platinum-based chemotherapy or PD-1 / L1 inhibitor monotherapy or combination therapy; and metastatic nasopharyngeal carcinoma that has previously received platinum-based chemotherapy or PD-1 / L1 inhibitor monotherapy or combination therapy.

[0082] In a preferred embodiment, the EGFR-sensitive mutations include EGFR exon 19 deletion and / or L858R mutation; the KRAS mutations include exon 2, 3, and 4 mutations; and the NRAS mutations include exon 2, 3, and 4 mutations.

[0083] In a preferred embodiment, the cancer is locally advanced or metastatic NSCLC with EGFR-sensitive mutations that has undergone systemic treatment with EGFR TKIs in an advanced stage, wherein the EGFR-sensitive mutations include EGFR exon 19 deletion and / or L858R mutation.

[0084] In a preferred embodiment, the cancer is EGFR wild-type metastatic NSCLC that has previously received platinum-based chemotherapy or PD-1 / L1 inhibitor monotherapy or combination therapy.

[0085] In a preferred embodiment, the cancer is recurrent or metastatic head and neck squamous cell carcinoma that has previously received platinum-based chemotherapy or PD-1 / L1 inhibitor monotherapy or combination therapy.

[0086] In a preferred embodiment, the cancer is a recurrent or metastatic colorectal adenocarcinoma that has failed or is intolerant to standard treatment and does not carry a KRAS mutation, NRAS mutation, or BRAF V600E mutation. The KRAS mutation includes exon 2, 3, and 4 mutations; the NRAS mutation includes exon 2, 3, and 4 mutations.

[0087] In a preferred embodiment, the cancer is a recurrent or metastatic squamous cell carcinoma of the oral cavity, oropharynx, hypopharynx, or larynx that has previously received platinum-based chemotherapy or PD-1 / L1 inhibitor monotherapy or combination therapy.

[0088] In a preferred embodiment, the cancer is metastatic nasopharyngeal carcinoma that has previously received platinum-based chemotherapy or PD-1 / L1 inhibitor monotherapy or combination therapy.

[0089] The anti-EGFR / c-Met antibody-drug conjugate disclosed herein possesses a series of superior properties: i) specific binding to EGFR and c-MET; ii) excellent endocytic activity in tumor cells expressing EGFR; and iii) superior in vitro and in vivo anticancer activity in tumor cells with different EGFR and c-MET expression levels. Specifically, the anti-EGFR / c-Met antibody of this disclosure, when conjugated with a cytotoxic drug, exhibits superior anticancer efficacy compared to the cytotoxic drug and is superior to existing conjugates of antibodies targeting the same target and cytotoxic drugs. Furthermore, the anti-EGFR / c-Met antibody-drug conjugate of this disclosure is superior to other cytotoxic drug-conjugated ADCs. Therefore, it can be used to treat cancer or to develop drugs for treating cancer.

[0090] the term

[0091] To facilitate understanding of this disclosure, certain technical and scientific terms are specifically defined below. Unless otherwise expressly defined herein, all other technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art to which this disclosure pertains.

[0092] The term "antibody-drug conjugate" refers to an antibody linked to a biologically active drug via a stable linker. In this disclosure, "antibody-drug conjugate" refers to a monoclonal antibody or antibody fragment linked to a biologically active toxic drug via a stable linker.

[0093] The terms "antibody-drug conjugate," "antibody-drug conjugate," and "ADC" have the same meaning: they refer to drugs formed from antibodies, cytotoxic drugs, and linkers that connect antibodies and cytotoxic drugs.

[0094] The term "antibody" refers to immunoglobulin, a tetrapeptide chain structure composed of two identical heavy chains and two identical light chains linked by interchain disulfide bonds. The amino acid composition and sequence of the constant region of the heavy chain of immunoglobulins differ, thus their antigenicity also differs. Based on this, immunoglobulins can be divided into five classes, or isotypes of immunoglobulins: IgM, IgD, IgG, IgA, and IgE, with their corresponding heavy chains being μ, δ, γ, α, and ε chains, respectively. Within the same class of Ig, differences in the amino acid composition of the hinge region and the number and position of disulfide bonds in the heavy chain can further divide them into different subclasses; for example, IgG can be divided into IgG1, IgG2, IgG3, and IgG4. The light chains are classified as κ or λ chains based on differences in the constant region. Each of the five classes of Ig can have either a κ chain or a λ chain.

[0095] The approximately 110 amino acids near the N-terminus of both the antibody heavy and light chains exhibit significant sequence variation and are termed the variable region (Fv region); the remaining amino acid sequences near the C-terminus are relatively stable and constitute the constant region. The variable region includes three hypervariable regions (HVRs) and four relatively conserved backbone regions (FRs). The three hypervariable regions determine the antibody's specificity and are also known as complementarity-determining regions (CDRs). Each light chain variable region (LCVR) and heavy chain variable region (HCVR) consists of three CDRs and four FRs, arranged from the amino terminus to the carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The three CDRs of the light chain refer to LCDR1, LCDR2, and LCDR3; the three CDRs of the heavy chain refer to HCDR1, HCDR2, and HCDR3.

[0096] In this disclosure, the amino acid sequences of the aforementioned CDRs are all derived according to the Kabat definition rules. However, it is well known to those skilled in the art that antibody CDRs can be defined in various ways, such as Chothia et al. (1989) Nature 342:877-883, Al-Lazikani et al., “Standard conformations for the canonical structures of immunoglobulins”, Journal of Molecular Biology, 273, 927-948 (1997)), Kabat et al. (1987) based on antibody sequence variability, AbM (University of Bath), Contact (University College London), the international ImMunoGeneTics database (IMGT) (imgt.cines.fr / ), and the North CDR definition based on affinity propagation clustering using a large number of crystal structures. Those skilled in the art will understand that, unless otherwise specified, the terms "CDR" and "complementarity-determining region" for a given antibody or its region (e.g., variable region) should be understood to encompass complementarity-determining regions defined as described above by any of the known schemes described in this invention. While the scope of protection claimed by this invention is based on the sequence shown in the Kabat definition rules, amino acid sequences corresponding to other CDR definition rules should also fall within the scope of protection of this invention.

[0097] The term “antigen-binding fragment” refers to one or more fragments of an antibody that maintain the ability to specifically bind to an antigen. It has been shown that fragments of full-length antibodies can be used for antigen-binding function. Examples of binding fragments included in “antigen-binding fragments” include (i) Fab fragments, monovalent fragments consisting of VL, VH, CL, and CH1 domains; (ii) F(ab')2 fragments, bivalent fragments comprising two Fab fragments linked by disulfide bridges on hinge regions; (iii) Fd fragments consisting of VH and CH1 domains; (iv) Fv fragments consisting of VH and VL domains of a single arm of the antibody; (v) single-domain or dAb fragments (Ward et al., (1989) Nature 341: 544-546) consisting of a VH domain; and (vi) separate complementarity-determining regions (CDRs) or (vii) combinations of two or more separate CDRs optionally linked by synthetic linkers.

[0098] The term "bispecific" refers to an antibody that specifically binds to two different antigens or two different epitopes within the same antigen. Bispecific antibodies may be cross-reactive to other related antigens, for example, to the same antigen from other species, such as humans or monkeys, such as cynomolgus monkeys or chimpanzees, or they may bind to epitopes shared between two or more different antigens.

[0099] The term "multispecific" refers to an antibody that specifically binds to two or more different antigens or two or more different epitopes within the same antigen. Multispecific antibodies may exhibit cross-reactivity with other related antigens, for example, cross-reactivity with the same antigen from other species, such as humans or monkeys, such as macaques or chimpanzees, or they may bind to epitopes shared between two or more different antigens.

[0100] The term "agonist" refers to a molecule that, when bound to a cellular protein, induces at least one response or activity induced by the protein's natural ligand. A molecule is an agonist when at least one response or activity is induced to a degree at least 20%, 30%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% greater than that induced in the absence of an agonist (e.g., a negative control), or when the induction is statistically significant compared to the induction in the absence of an agonist.

[0101] The terms "antagonist" or "inhibitor" refer to a molecule that, when bound to a cellular protein, inhibits at least one reaction or activity induced by the protein's natural ligand. A molecule is an antagonist when at least one reaction or activity is inhibited by at least 20%, 30%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% more than when at least one reaction or activity is inhibited in the absence of an antagonist (e.g., a negative control), or when the inhibition is statistically significant compared to the inhibition in the absence of an antagonist.

[0102] The term "drug loading" refers to the average number of cytotoxic drugs loaded onto each ligand in a molecule of formula (1), and can also be expressed as the ratio of drug amount to antibody amount. The drug loading range can be 0-12, preferably 1-10, cytotoxic drugs (D) linked to each antibody (Pc). In embodiments of this disclosure, the drug loading is expressed as n, also known as the DAR value, exemplarily the average of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. The average number of drugs per ADC molecule after the coupling reaction can be identified using conventional methods such as UV / visible spectroscopy, mass spectrometry, ELISA assays, and HPLC characterization.

[0103] The term "combination" or "drug combination" defines a fixed combination of dosage units or a kit of drugs for combined administration, wherein the therapeutic agents can be administered simultaneously or separately at time intervals, so that the therapeutic agents can exhibit a synergistic effect, such as a synergistic effect.

[0104] The term “pharmaceutically acceptable” is defined herein as those compounds, materials, biological agents, compositions and / or dosage forms that are suitable for contact with the tissues of a subject (such as mammals or humans) within the limits of reasonable medical judgment, without excessive toxicity, irritation, allergic reactions and other problematic complications, and with a reasonable benefit / risk ratio.

[0105] The term "pharmaceutical composition" refers to a product that optionally contains one or more active ingredients (e.g., antibodies, ADCs) in a specific amount, and any product directly or indirectly produced by combining one or more active ingredients in optional specific amounts. The different active ingredients in a pharmaceutical composition may be administered independently as separate formulations, including synergistically, either simultaneously or at different times. In this disclosure, "pharmaceutical composition" and "formulation" are not mutually exclusive.

[0106] The term "identity" refers to the sequence matching between two polypeptides or two nucleic acids. Two compared sequences are considered identical at that position when a position is occupied by the same base or amino acid monomeric subunit (e.g., a position in each of two DNA molecules is occupied by adenine, or a position in each of two polypeptides is occupied by lysine). The "percentage identity" between two sequences is a function of the number of matching positions shared by the two sequences divided by the number of positions compared, multiplied by 100.

[0107] The term "treatment" means administering an oral or topical therapeutic agent, such as a composition comprising any of the compounds disclosed herein, to a patient who has symptoms of one or more diseases, and the therapeutic agent is known to have a therapeutic effect on these symptoms. Typically, a therapeutic agent is administered in a treated patient or population in an amount that effectively relieves symptoms of one or more diseases to induce the regression of such symptoms or inhibit their progression to any clinically measurable extent. The amount of a therapeutic agent that effectively relieves any specific disease symptom (also referred to as a "therapeuticly effective amount") can vary depending on a variety of factors, such as the patient's disease state, age, and weight, and the drug's ability to produce the desired therapeutic effect in the patient. Whether the disease symptoms have been relieved can be evaluated using any clinical testing method commonly used by a physician or other healthcare professional to assess the severity or progression of the symptoms. Although the embodiments of this disclosure (e.g., treatment methods or products) may be ineffective in alleviating symptoms of each target disease, they should reduce symptoms of the target disease in a statistically significant number of patients, as determined by any statistical test known in the art, such as the Student t-test, chi-square test, U-test according to Mann and Whitney, Kruskal-Wallis test (H-test), Jonckheere-Terpstra test, and Wilcoxon test.

[0108] The term "effective dose" or "therapeutic effective dose" for a combination of therapeutic agents is the amount that is sufficient to provide a significant improvement in clinically observable signs and symptoms compared to baseline for the cancer treated by the combination.

[0109] The terms “cancer” and “tumor” are used interchangeably, referring to a large category of diseases characterized by the uncontrolled growth of abnormal cells in the body.

[0110] "Cancer expressing EGFR and / or c-Met" refers to cancer with detectable EGFR and / or c-Met expression or with EGFR and / or c-Met mutations or amplifications. EGFR and / or c-Met expression, amplification, and mutation status can be detected using known methods such as sequencing, fluorescence in situ hybridization, immunohistochemistry, flow cytometry, or Western blotting using tumor biopsy or blood samples. Expression can also be detected by sequencing circulating tumor DNA (ctDNA).

[0111] The term "subject" refers to a mammal, such as a primate, like a human. In some implementations, the subject (e.g., a human) has a tumor, or is at risk of having the aforementioned disease.

[0112] The term "effective amount" refers to an amount sufficient to achieve, or at least partially achieve, the desired effect. For example, an effective amount for preventing disease (e.g., cancer) is an amount sufficient to prevent, stop, or delay the onset of disease (e.g., cancer); an effective amount for treating disease is an amount sufficient to cure or at least partially stop the disease and its complications in a patient already suffering from the disease. Determining such an effective amount is entirely within the capabilities of those skilled in the art. For example, an effective amount for therapeutic purposes will depend on the severity of the disease to be treated, the overall state of the patient's own immune system, the patient's general characteristics such as age, weight, and sex, the manner of administration of the drug, and other concurrent treatments, etc.

[0113] The terms “including,” “comprising,” “having,” “containing,” or “involving,” and their other variations herein, are inclusive or open-ended and do not exclude other unlisted elements or method steps. Attached Figure Description

[0114] Figure 1: Schematic diagram of the dose exploration phase trial design.

[0115] Figure 2: Schematic diagram of the dose expansion phase experimental design. Detailed Implementation

[0116] The following embodiments are used to further describe the present invention, but these embodiments are not intended to limit the scope of the present invention.

[0117] Experimental methods not specifically described in the embodiments or test examples of this disclosure are generally performed under conventional conditions or as recommended by the raw material or product manufacturer. See Sambrook et al., Molecular Cloning, Laboratory Manual, Cold Spring Harbor Laboratory; Methods in Modern Molecular Biology, Ausubel et al., Greene Publishing Association, Wiley Interscience, NY. Reagents not specifically named are commercially available, conventional reagents.

[0118] Example 1. Preparation of anti-EGFR / c-Met antibody-drug conjugates

[0119] Preparation of bispecific antibodies

[0120] The bispecific antibody EGFRxc-MET-136Afu (named BsAb-1 in this application) was prepared according to the method disclosed in Example 6 of patent WO2023186092. The entire contents of the aforementioned patent WO2023186092 are incorporated herein by reference.

[0121] The bispecific antibody BsAb-1 is composed of EGFR heavy chain 1, c-Met heavy chain 1, EGFR light chain 1, and c-Met light chain 1 (SEQ ID NO: 25-28). The CDR region, variable region, constant region, and full-length amino acid sequence of the bispecific antibody are shown in Table 1. The amino acid residues in the CDR region are numbered based on the Kabat numbering system.

[0122] Table 1. Sequence information of anti-EGFR / c-Met antibodies

[0123] Preparation of drug loading

[0124] Anti-EGFR / c-Met antibody-drug conjugates were prepared by conjugating compounds C and D with anti-EGFR / c-Met antibodies. Compound C was prepared using the method disclosed in patent WO2020063673, and compound D was prepared using the method disclosed in patent WO2022161385. The structural information of compounds C and D is as follows:

[0125] Compounds C and D enter tumor cells via the endocytosis effect of ADCs to release the toxin molecule compound 2-A. Compound 2-A, as an effective inhibitor of DNA topoisomerase I, is disclosed in patent WO2021190480, and is prepared according to the method disclosed in that patent application.

[0126] Conjugation of bispecific antibodies to drug payload

[0127] The bispecific antibody BsAb-1 (5 mg / mL PBS, pH 7.4) was treated with a sufficient molar equivalent of TCEP (10 mM) at 37°C for 1 hour. Isotype mAb1 and mAb2 served as negative controls, while Ref1 and Ref2 were positive control antibodies. Ref1 antibody is cited from patent WO2023083846, and Ref2 antibody is cited from patent WO2014081954. Specifically, as described in patent WO2023083846, Ref1 antibody was simultaneously conjugated with the drug payload SG3932 to construct a positive control ADC.

[0128] A sufficient molar equivalent of the drug load (such as compound C or compound D mentioned above) in DMSO was added to PBS containing the reducing antibody. After incubation for 1 h, the reaction mixture was purified by size exclusion chromatography (SEC) to separate the ADC and the free unconjugated drug load. The ADC structure formed by conjugating the bispecific antibody with compound C is shown in general formula (ADC-A), and the ADC structure formed by conjugating it with compound D is shown in general formula (ADC-B).

[0129] The DAR values ​​(y) of the antibody-drug conjugates were determined using a TOF LC / MS system, and the average DAR values ​​are shown in Table 2. The average DAR values ​​of the ADC conjugated with compound C were approximately 4 and 6. The average DAR values ​​of the ADC conjugated with compound D were approximately 4, 6, and 8.

[0130] Table 2: Conjugation results of bispecific antibodies and drug loading

[0131] Example 2. Internalization efficiency of anti-EGFR / c-Met antibody-drug conjugate

[0132] The pHrodo™ iFL reagent was used to label the test substances with a pH-dependent fluorescent dye for flow cytometry detection to evaluate the differences in endocytosis of antibody-drug conjugates, naked antibodies, and EGFR-c-Met ADCs on tumor cells expressing different levels of EGFR and c-Met.

[0133] HCC827 and MDA-MB-468 monolayers in logarithmic growth phase were rinsed once with 10 mL of PBS, incubated at 37°C for 3 min with 6 mL of 0.25% Trypsin-EDTA cell digestion buffer, and then added 15 mL of complete culture medium to stop digestion. Cells were transferred to centrifuge tubes and centrifuged at 1000 rpm for 5 min. After rinsing once with 10 mL of PBS, cells were collected by centrifugation and resuspended in 5 mL of complete culture medium. Cell density was measured using a cell counter and adjusted to 2 × 10⁶ cells / mL. 6Cells / mL. Dilute the ADC and naked antibody stock solution to 40 nM or 80 nM with complete culture medium. Then dilute the pHrodo™ iFL reagent to 120 nM or 240 nM with complete culture medium. Finally, mix the test sample and pHrodo™ iFL reagent dilution at a molar ratio of 1:3 and a volume ratio of 1:1, and incubate at room temperature for 10 min. Add 50 μL / well of the test sample labeled with pHrodo™ iFL fluorescent dye to the cell plate using a 12-channel manual pipette, mix gently, and incubate at 37°C. The final concentration of the test sample is 10 nM, and the final concentration of the pHrodo™ iFL reagent is 30 nM.

[0134] Cells were removed from the 37°C incubator at three time points: 0.5h, 2h, and 4h. The cells were resuspended by pipetting and then placed on a flow cytometer for analysis. The data were imported into the Flowjo analysis software to obtain the average fluorescence signal intensity (MFI) of different sample wells. The MFI of the cells at each time point was compared with that of the cell wells (Baseline) with pHrodo™ iFL reagent added individually to obtain the proportion of MFI changes of different cells at each time point.

[0135] Table 3. Changes in mean fluorescence signal (MFI) of test substances at different time points

[0136] Example 3. In vitro cell killing assay of anti-EGFR / c-Met antibody-drug conjugate

[0137] The inhibitory effect of ADC drugs on tumor cell growth was detected using an in vitro cell killing assay. Different tumor cells in the logarithmic growth phase were collected and evenly distributed at a density of 2000 cells / well in 96-well plates. After overnight incubation, ADC was added to each well to achieve a final concentration range of 0.003 nM to 200 nM. After incubation for 4–7 days, the inhibitory effect was assessed using... The 2.0 assay kit was used to homogenize and detect the number of viable cells in cultured cells. The IC50 was obtained by curve fitting using GraphPad Prism 9 software. 50 Value, ADC for in vitro IC50 values ​​of tumor cell lines expressing different levels of EGFR / c-Met antigen 50 The values ​​are as follows.

[0138] ADCs can significantly inhibit tumor cells expressing EGFR / c-Met antigens from different tissue sources, and their efficacy is superior to that of control or Yangshen ADCs.

[0139] Table 4. IC50 of ADCs for inhibiting the proliferation of different tumor cells 50 Value (nM) Note: "-" indicates not detected. "NA" indicates not applicable.

[0140] use A chemiluminescent cell viability assay kit was used to detect the inhibitory effect of drugs on tumor cell proliferation and growth. A 10x working solution of the test drug was prepared by uniformly diluting the stock solution of the test drug to a maximum concentration of 1, 3, 10, or 20 μM using serum-free medium. Then, it was serially diluted 3-fold or 4-fold, resulting in a total of 9 concentrations. The prepared 10x test drug dilutions were added sequentially to 96-well cell culture plates of different cell types, 20 μL per well, resulting in a final test drug concentration of 2000 nM–1.526 pM. Each concentration was replicated. For the culture medium control wells, 20 μL of culture medium was added to each well. The cell culture plates were incubated at 37°C in a 5% CO2 incubator for 6 days.

[0141] After incubating the test drug with the cells for a period of time, remove the cell culture plate and equilibrate it to room temperature, then add 50 μL / well. The reagents were used to induce cell lysis and ATP release by shaking on a shaker for 10 min. The culture plate was then incubated at room temperature in the dark for 10 min. The chemiluminescence signal value was detected using a microplate reader at 1000 ms / well. The inhibition rate (%) was calculated from the chemiluminescence signal value: Inhibition rate (%) = (1 - sample well signal value / average signal value of solvent control wells) x 100%. Based on the inhibition rate of each concentration of the test drug, the IC50 was obtained by curve fitting using GraphPad Prism9 software. 50 value.

[0142] Both the tested ADC and compound 2-A significantly inhibited tumor cells expressing EGFR / cMet antigen from different tissue sources, and the inhibitory effect of ADC-12 was better than that of the control ADC and the cytotoxic drug 2-A.

[0143] Table 5. IC50 of ADCs for inhibiting the proliferation of different tumor cells 50 Value (nM)

[0144] Example 4. In vivo efficacy study of ADC in a nude mouse subcutaneous xenograft tumor model

[0145] 1. NCI-H1975 cell xenograft (CDX) model

[0146] NCI-H1975 tumor cells were subcutaneously implanted into Balb / C nude mice. When the tumor volume reached approximately 150-200 mm², the tumor cells were... 3 At that time, transplanted mice were randomly divided into 4 groups (n=5 per group). Each group was treated with control, ADC-08, and ADC-09, respectively. Mice were intravenously injected with 3 mpk of ADC, QWx3. The mean tumor growth inhibition (TGI) was calculated using the following formula:

[0147] TGI(%) = ((1-(mean tumor volume at the end of treatment group - mean tumor volume at the time of grouping)) / (mean tumor volume at the end of treatment in solvent control group - mean tumor volume at the time of grouping in solvent control group)) x 100%.

[0148] The results are shown in Table 6. Compared with the control group, the tumor growth of mice treated with ADC showed statistically significant differences, leading to tumor regression. The tumor growth of mice treated with ADC-08 and ADC-09 showed statistically significant differences compared with the control mice, with TGI values ​​of 103.2% and 104.0% on day 16, respectively.

[0149] Table 6. Evaluation of antitumor efficacy in the NCI-H1975 xenograft tumor model

[0150] In another experiment, NCI-H1975 tumor-bearing mice were randomly divided into 7 groups, with 5 mice in each group. Each group was treated with a single intravenous injection of either control ADC-11, ADC-01, or ADC-02 at two different dose levels. Compared with the control group, the tumor growth in ADC-treated mice was statistically different, leading to tumor regression. Compared with ADC-11, ADC-01 and ADC-02 significantly delayed tumor growth. ADC-02 was more effective than ADC-01. The results are shown in Table 7.

[0151] Table 7. Evaluation of antitumor efficacy in the NCI-H1975 xenograft tumor model

[0152] In another experiment, NCI-H1975 tumor-bearing mice were randomly divided into 12 groups, with 5 mice in each group. Each group was treated with a single intravenous injection of control, BsAb-1 ADC, Ref1 ADC, or BsAb-1, respectively. Compared with ADC-03, ADC-04, and BsAb-1, ADC-01 and ADC-02 significantly delayed tumor growth. The results are shown in Table 8.

[0153] Table 8. Evaluation of antitumor efficacy in the NCI-H1975 xenograft tumor model

[0154] 2. NCI-H1975-EGFR C797S cell xenograft (CDX) model

[0155] NCI-H1975 (EGFR C797S) tumor cells were subcutaneously implanted into NOD / SCID mice. When the tumor volume reached approximately 150-200 mm², the tumor cells were implanted into NOD / SCID mice. 3 At that time, transplanted mice were randomly divided into 4 groups (n=5 per group). Each group was treated with either a control or ADC-08. Mice were intravenously injected with ADC 3 mpk, QW×2. The mean tumor growth inhibition (TGI) was calculated using the following formula:

[0156] TGI(%) = ((1-(mean tumor volume at the end of treatment group - mean tumor volume at the time of grouping)) / (mean tumor volume at the end of treatment in solvent control group - mean tumor volume at the time of grouping in solvent control group)) x 100%.

[0157] The results are shown in Table 9. The tumor growth of mice treated with ADC-08 was statistically different from that in the Vehicle group, with a TGI value of 108.55% on day 28.

[0158] Table 9. Evaluation of antitumor efficacy in the NCI-H1975-EGFR C797S xenograft tumor model

[0159] 3. NCI-H441 xenograft tumor model

[0160] NCI-H441 tumor cells were subcutaneously implanted into BALB / c mice. When the tumor volume reached approximately 100-200 mm², the tumor cells were implanted. 3 At the time of transplantation, transplanted mice were randomly divided into 4 groups (n=6 per group). Each group was treated with BsAb-1 and compounds 2-A, ADC-12, and ADC-13, respectively. Mice were intravenously injected with ADC (1, 3, and 6 mg / kg) or a single dose of BsAb-1 + compound 2-A (mAb + drug_6 + 0.086 mg / kg), respectively. The mean tumor growth inhibition (TGI) was calculated using the following formula:

[0161] TGI(%) = ((1-(mean tumor volume at the end of treatment group - mean tumor volume at the time of grouping)) / (mean tumor volume at the end of treatment in solvent control group - mean tumor volume at the time of grouping in solvent control group)) x 100%.

[0162] Single intravenous administration of ADC-12 at doses of 1, 3, and 6 mg / kg all demonstrated significant tumor-suppressive effects, exhibiting a favorable dose-response relationship. At the same dose, ADC-12 showed superior tumor-suppressive activity compared to naked anti-BsAb-1 and ADC-13. Animals in all ADC-12 dose groups remained in good condition with no abnormalities.

[0163] Table 10. Evaluation of antitumor efficacy in the NCI-H441 xenograft tumor model

[0164] 4. SNU-1 xenograft tumor model

[0165] SNU-1 tumor cells were subcutaneously implanted into BALB / c mice. When the tumor volume reached approximately 150-200 mm², the tumor cells were implanted. 3At that time, transplanted mice were randomly divided into 4 groups (n=7 per group). Each group was treated with BsAb-1 and compounds 2-A, ADC-12, and ADC-13, respectively. Mice were intravenously injected with single doses of ADC (1, 3, and 9 mg / kg) and BsAb-1 + compound 2-A (mAb + drug 9 + 0.128 mg / kg), respectively. The mean tumor growth inhibition (TGI) was calculated using the following formula:

[0166] TGI(%) = ((1-(mean tumor volume at the end of treatment group - mean tumor volume at the time of grouping)) / (mean tumor volume at the end of treatment in solvent control group - mean tumor volume at the time of grouping in solvent control group)) x 100%.

[0167] Single intravenous injections of ADC-12 at doses of 1, 3, and 9 mg / kg all demonstrated significant tumor-suppressive effects, exhibiting a favorable dose-response relationship. At the same dose, ADC-12 showed significantly superior tumor-suppressive effects compared to naked anti-BsAb-1 and Yangshen ADC. All animals in the groups were in good condition with no abnormalities.

[0168] Table 11. Evaluation of antitumor efficacy in the SNU-1 xenograft tumor model

[0169] 5. HCT-116 xenograft tumor model

[0170] HCT-116 tumor cells were subcutaneously implanted into BALB / c mice. When the tumor volume reached approximately 100-200 mm², the cells were cultured. 3 At the time of transplantation, transplanted mice were randomly divided into 4 groups (n=7 per group). Each group was treated with BsAb-1 and compounds 2-A, ADC-12, and ADC-13, respectively. Mice were intravenously injected with ADC (1, 3, 8 mg / kg) and a single dose of BsAb-1 + compound 2-A (mAb + drug 8 + 0.114 mg / kg), respectively. The mean tumor growth inhibition (TGI) was calculated using the following formula:

[0171] TGI(%) = ((1-(mean tumor volume at the end of treatment group - mean tumor volume at the time of grouping)) / (mean tumor volume at the end of treatment in solvent control group - mean tumor volume at the time of grouping in solvent control group)) x 100%.

[0172] Single intravenous injections of ADC-12 at doses of 1, 3, and 8 mg / kg all demonstrated significant tumor-suppressive effects, exhibiting a favorable dose-response relationship. At the same dose, ADC-12 showed significantly superior tumor-suppressive effects compared to naked anti-BsAb-1 and Yangshen ADC. All animals in the groups were in good condition with no abnormalities.

[0173] Table 12. Evaluation of antitumor efficacy in the HCT-116 xenograft tumor model

[0174] 6. Detroit 562 xenograft tumor model

[0175] Detroit562 tumor cells were subcutaneously implanted into BALB / c mice. When the tumor volume reached approximately 100-200 mm², the tumor cells were successfully implanted. 3 At the time of transplantation, transplanted mice were randomly divided into 4 groups (n=6 per group). Each group was treated with BsAb-1 and compounds 2-A, ADC-12, and ADC-13, respectively. Mice were intravenously injected with ADC (0.3, 1, and 3 mg / kg) and a single dose of BsAb-1 + compound 2-A (mAb + drug 3 + 0.043 mg / kg), respectively. The mean tumor growth inhibition (TGI) was calculated using the following formula:

[0176] TGI(%) = ((1-(mean tumor volume at the end of treatment group - mean tumor volume at the time of grouping)) / (mean tumor volume at the end of treatment in solvent control group - mean tumor volume at the time of grouping in solvent control group)) x 100%.

[0177] Single intravenous injections of ADC-12 at doses of 0.3, 1, and 3 mg / kg all demonstrated significant tumor-suppressive effects, exhibiting a favorable dose-response relationship. At the same dose, ADC-12 showed significantly superior tumor-suppressive effects compared to naked anti-BsAb-1 and Yangshen ADC. All animals in the groups were in good condition with no abnormalities.

[0178] Table 13. Evaluation of the antitumor efficacy of Detroit 562 xenograft tumor model

[0179] Example 5. Clinical trial of anti-EGFR / c-Met antibody-drug conjugate for the treatment of advanced solid tumors

[0180] I. Research Objectives

[0181] Main research objective:

[0182] a) Evaluate the efficacy of intravenous administration of anti-EGFR / c-Met antibody-drug conjugates in patients with advanced solid tumors. b) Evaluate the safety and tolerability of intravenous administration of anti-EGFR / c-Met antibody-drug conjugates in patients with advanced solid tumors.

[0183] Secondary research objective:

[0184] a) Evaluate other efficacy endpoints of intravenous administration of anti-EGFR / c-Met antibody-drug conjugates in patients with advanced solid tumors. b) Evaluate the pharmacokinetic (PK) characteristics of intravenous administration of anti-EGFR / c-Met antibody-drug conjugates in patients with advanced solid tumors. c) Evaluate the immunogenicity of intravenous administration of anti-EGFR / c-Met antibody-drug conjugates in patients with advanced solid tumors.

[0185] II. Name of the test drug:

[0186] Anti-EGFR / c-Met antibody-drug conjugate

[0187] The anti-EGFR / c-Met bispecific antibody-drug conjugate is ADC-12, the preparation of which is described in Example 1. Dosage form: sterile lyophilized powder for injection, specification: 100mg / vial, provided in a combination packaging form of sterile borosilicate glass vial, film-coated butyl rubber stopper for injecting lyophilized sterile powder, and aluminum-plastic combination cap for antibiotic vials.

[0188] III. Target Audience:

[0189] 1. Males or females who are 18 years of age or older (≥18 years old)

[0190] 2. The dose exploration phase requires inclusion of pathologically confirmed advanced solid tumors that have failed adequate standard treatment or are intolerant to standard treatment, including but not limited to: NSCLC, HNSCC, or CRC; the dose expansion phase requires inclusion of pathologically confirmed advanced solid tumors that have failed adequate standard treatment or are intolerant to standard treatment. The criteria for each cohort are as follows:

[0191] Cohort 1: Histologically or cytologically confirmed locally advanced or metastatic NSCLC that has failed or is intolerant of adequate standard therapy, and is confirmed to have EGFR-sensitive mutations. Systemic therapy for advanced stages is limited to EGFR TKIs (combination therapy with bevacizumab or its biosimilars concurrent with the EGFR TKI may be accepted).

[0192] Cohort 2: Metastatic NSCLC confirmed by histology or cytology, which has failed or is intolerant of adequate standard therapy, is confirmed to be EGFR wild-type and has no other known manipulable gene alterations (no detected or detected mutations for which no corresponding antitumor drugs have been approved by the NMPA, excluding MET abnormalities such as exon 14 skipping, MET amplification determined by various methods, and MET overexpression). Patients have previously received platinum-based chemotherapy or PD-1 / L1 inhibitor therapy or combination therapy, but have received no more than two lines of systemic therapy for metastatic disease.

[0193] Cohort 3: Histologically or cytologically confirmed unresectable recurrent or metastatic colorectal adenocarcinoma that has failed adequate standard therapy or is intolerant to standard therapy, and is free from KRAS mutations (exon 2, 3, 4 mutations), NRAS mutations (exon 2, 3, 4 mutations), or BRAF V600E mutations. Patients with locally advanced / metastatic CRC have received no more than three lines of prior systemic therapy.

[0194] Cohort 4: Histologically confirmed recurrent or metastatic squamous cell carcinoma of the oral cavity, oropharynx, hypopharynx, or larynx that has failed or is intolerant of adequate standard treatment. Patients have previously received platinum-based chemotherapy, PD-1 / L1 inhibitor therapy, or combination therapy, but have not received more than two lines of systemic therapy for metastatic disease.

[0195] Cohort 5: Metastatic nasopharyngeal carcinoma confirmed by histology or cytology, which has failed adequate standard treatment or is intolerant to standard treatment. Patients have previously received platinum-based chemotherapy, PD-1 / L1 inhibitor therapy, or combination therapy, but have received no more than two lines of systemic therapy for metastatic disease.

[0196] Cohort 6: Histologically confirmed recurrent or metastatic squamous cell carcinoma of the esophagus or gastroesophageal junction that has failed adequate standard treatment or is intolerant to standard treatment. Prior systemic therapy for locally advanced / metastatic tumors should not exceed two lines.

[0197] Cohort 7: Other advanced solid tumors that have been histologically or cytologically confirmed and have failed adequate standard treatment or are intolerant to standard treatment.

[0198] 3. According to RECIST v1.1, participants must have at least one target lesion as assessed by the investigator. A target lesion is defined as a measurable lesion that has not undergone local treatment such as irradiation, or has clearly progressed after local treatment, with a longest diameter ≥10 mm at baseline (or a shortest diameter ≥15 mm for lymph nodes). Participants with only brain and / or bone lesions as target lesions will not be accepted.

[0199] 4. Good bone marrow reserves or good liver and kidney function.

[0200] 5. The ECOGPS score was 0–1, and there was no worsening in the two weeks prior to the first dose.

[0201] 6. Minimum expected survival is greater than 12 weeks.

[0202] 7. Voluntarily participate in this clinical trial, understand the research procedures, and be able to sign an informed consent form in writing.

[0203] IV. Dosing regimen:

[0204] The anti-EGFR / c-Met antibody-drug conjugate is administered via intravenous infusion. The total dose of the anti-EGFR / c-Met antibody-drug conjugate is calculated based on the participant's weight before each dose. The recommended duration of the initial intravenous infusion is 90 ± 10 minutes (excluding the flushing phase, which should be completed within 30 minutes of administration). Subsequent infusions may be adjusted based on the participant's tolerance, but should not be less than 30 minutes (excluding the flushing phase, which should be completed within 30 minutes of administration).

[0205] The dosing frequency for the anti-EGFR / c-Met antibody-drug conjugate is Q3W, with one treatment cycle lasting 21 days, continuing until objective disease progression (except for continued treatment after disease progression) or other criteria for discontinuing study treatment as specified in the protocol are met. Based on the obtained safety, pharmacokinetic, efficacy, and immunogenicity data, the dosing frequency may be adjusted (e.g., every Q2W). For example, if adjusted to Q2W, with one treatment cycle lasting 28 days, dosing once every 14 days.

[0206] The dose escalation phase employs a BOIN design. The starting dose of the anti-EGFR / c-Met antibody-drug conjugate is 3 mg / kg Q3W, with four dose escalations (3 mg / kg, 6 mg / kg, 8 mg / kg, and 10 mg / kg). The Safety Review Committee (SRC) will collaboratively discuss and decide whether additional dose groups (higher or intermediate doses, such as 5.0 mg / kg Q3W, 4.8 mg / kg Q3W, 4.5 mg / kg Q3W, 4.2 mg / kg Q3W, 4.0 mg / kg Q3W, 7 mg / kg Q3W, 9 mg / kg Q3W, etc.) are needed based on obtained safety, tolerability, pharmacokinetic, efficacy, and immunogenicity information. Exposure simulations will be performed based on the confirmed safe Q3W dose to determine the exploratory dose for Q2W (e.g., 4 mg / kg Q2W, 5 mg / kg Q2W, 6 mg / kg Q2W, 7 mg / kg Q2W, and 8 mg / kg Q2W, etc.).

[0207] The dose expansion phase will be conducted using the dosing regimen established in the dose escalation phase studies. Each cohort will be planned to enroll 30–50 participants at each dose level.

[0208] V. Study endpoint:

[0209] Primary study endpoint:

[0210] a) Based on ORR assessed by researchers according to RECIST v1.1 standards.

[0211] b) Based on investigator-assessed DCR, DoR, and PFS according to RECIST v1.1 criteria; overall survival (OS).

[0212] c) The maximum tolerated dose (MTD) or maximum applicable dose (MAD) of the intravenous anti-EGFR / c-Met antibody-drug conjugate.

[0213] Secondary study endpoints:

[0214] a) Incidence of adverse events (AEs), serious adverse events (SAEs), AEs leading to dose adjustment or permanent discontinuation of the drug, and specific laboratory abnormalities.

[0215] b) Blood drug concentrations of the anti-EGFR / c-Met antibody-drug conjugate and its components (including ADC, total antibody and free payload); pharmacokinetic parameters of the anti-EGFR / c-Met antibody-drug conjugate in humans.

[0216] c) The incidence of ADA.

[0217] VI. Research Results:

[0218] 6.1 Security Assessment

[0219] As of January 29, 2026, 54 patients were evaluable for safety. No DLT events were observed in any dose group, and no patients permanently discontinued the drug or died due to adverse events. Common adverse events related to the study drug included hematologic toxicity, infusion-related reactions, gastrointestinal toxicity, asthenia, alopecia, myalgia, and hepatotoxicity. Except for granulocytic toxicity (neutropenia) in the 8 mg / kg high-dose group, most other adverse events were grade 1-2 and were generally manageable and tolerable.

[0220] 6.2 Effectiveness Assessment

[0221] As of January 29, 2026, 25 patients with EGFR-mutant NSCLC in Cohort 1 who had previously received EGFR TKI therapy completed at least one tumor efficacy assessment across the three dose groups. Efficacy data are shown in Table 14.

[0222] Table 14. Summary of Objective Response Rate and Disease Control Rate

[0223] In cohort 2, patients with EGFR wild-type lung cancer (who had completed at least one tumor efficacy assessment) showed preliminary antitumor efficacy.

[0224] In summary, this study preliminarily validates the antitumor activity of the anti-EGFR / c-Met antibody-drug conjugate. The drug demonstrated clear and superior efficacy across all dosage groups, with a favorable safety profile; the nature and severity of adverse events were within expectations and were generally manageable and controllable.

Claims

1. Use of anti-EGFR / c-Met antibody-drug conjugates and their tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates or isotopic labels as shown in any one of (1)-(3) below: (1) Use in the preparation of drugs for treating cancer; (2) Used for the prevention and / or treatment of cancer; (3) For the prevention and / or treatment of a subject with cancer, wherein the anti-EGFR / c-Met antibody-drug conjugate and its tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates or isotopic labels are administered to the subject. in, The structures of the anti-EGFR / c-Met antibody-drug conjugates and their tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates, or isotope-labeled compounds are shown in general formula (1). in: BsAb is an anti-EGFR / c-Met antibody or its antigen-binding fragment, comprising: at least one antigen-binding module 1 that specifically binds to EGFR and at least one antigen-binding module 2 that specifically binds to c-Met; y is selected from decimals or integers from 1 to 12; -L- represents a connector that covalently links BsAb to D; D is a cytotoxic drug, an immunostimulant, or a radioactive isotope, preferably a DNA topoisomerase inhibitor, a tubulin inhibitor, a DNA damaging agent, antimetabolite, or an antitumor antibiotic; more preferably, camptothecin or its analogues or derivatives, erribulin or its analogues or derivatives, auristatin derivatives, maytansine or its analogues or derivatives, dolastatin or its derivatives, calicacin or pyrrolobenzodiazepine (PBD); further preferably, exatecan or its analogues or derivatives, erribulin or its analogues or derivatives, MMAE, MMAF, MMAD, SN-38, DM1, or DM4; Preferably, The antigen-binding module 1 includes a heavy chain variable region 1 (VH1) and a light chain variable region 1 (VL1), and the antigen-binding module 2 includes a heavy chain variable region 2 (VH2) and a light chain variable region 2 (VL2). in, i) The VH1 contains the amino acid sequences of the heavy chain complementarity-determining region (HCDR)1 (HCDR1), HCDR2, and HCDR3 as shown in SEQ ID NO:07, 08, and 09, respectively; and ii) The VL1 contains the amino acid sequences of the light chain complementarity-determining region (LCDR)1 (LCDR1), LCDR2 and LCDR3 as shown in SEQ ID NO:10, 11 and 12, respectively; iii) The VH2 comprises the amino acid sequences HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO:01, 02, and 03, respectively; and iv) The VL2 comprises the amino acid sequences LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO:04, 05 and 06, respectively; More preferably, the anti-EGFR / c-Met antibody or its antigen-binding fragment contains: i) The VH1 contains an amino acid sequence having at least 90%, 95%, or 99% sequence identity with SEQ ID NO: 15, and the VL1 contains an amino acid sequence having at least 90%, 95%, or 99% sequence identity with SEQ ID NO: 16; ii) The VH2 contains an amino acid sequence having at least 90%, 95%, or 99% sequence identity with SEQ ID NO: 13, and the VL2 contains an amino acid sequence having at least 90%, 95%, or 99% sequence identity with SEQ ID NO: 14; More preferably, i) The VH1 comprises the sequence shown in SEQ ID NO: 15, and the VL1 comprises the sequence shown in SEQ ID NO: 16; ii) The VH2 contains the sequence shown in SEQ ID NO: 13, and the VL2 contains the sequence shown in SEQ ID NO:

14.

2. The use according to claim 1, wherein the anti-EGFR / c-Met antibody or its antigen-binding fragment comprises: a) The first heavy chain (HC1), which contains the VH1 and HC1 heavy chain constant regions (HC1_CH). b) The second heavy chain (HC2), which contains the constant regions of VH2 and HC2 heavy chains (HC2_CH). c) The first light chain (LC1), which contains the constant regions of VL1 and LC1 light chains (LC1_CL), and d) The second light chain (LC2), which contains the constant regions of VL2 and LC2 light chains (LC2_CL); in, i) The HC1_CH and HC2_CH contain the IgG heavy chain constant region or a variant thereof, preferably the IgG1, IgG2, IgG3 or IgG4 heavy chain constant region or a variant thereof, more preferably the sequence as shown in SEQ ID NO: 17 or a variant thereof. ii) The LC1_CL and LC2_CL contain a k or λ light chain constant region or a variant thereof, preferably containing a k light chain constant region or a variant thereof, more preferably containing a sequence as shown in SEQ ID NO: 18 or a variant thereof; Wherein, the variant described in any one of i) and ii) has at least 80%, 85%, 90%, 95%, or 99% sequence identity with the sequence from which it is derived, or the variant has at most 20, 15, 10, 9, 8, 7, 6, or 5 amino acid substitutions, deletions, or additions compared with the sequence from which it is derived; Preferably, the HC1_CH and HC2_CH comprise sequences as shown in SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21 or SEQ ID NO: 22, and the LC1_CL and LC2_CL comprise sequences as shown in SEQ ID NO: 18, SEQ ID NO: 23, SEQ ID NO: 24; More preferably, the HC1_CH contains a sequence as shown in SEQ ID NO: 17, SEQ ID NO: 19, or SEQ ID NO: 21, the HC2_CH contains a sequence as shown in SEQ ID NO: 17, SEQ ID NO: 20, or SEQ ID NO: 22, the LC1_CL contains a sequence as shown in SEQ ID NO: 18, or SEQ ID NO: 23, and the LC2_CL contains a sequence as shown in SEQ ID NO: 18, or SEQ ID NO:

24.

3. The use according to claim 2, wherein HC1 comprises a sequence as shown in SEQ ID NO: 25 or SEQ ID NO: 29, HC2 comprises a sequence as shown in SEQ ID NO: 26 or SEQ ID NO: 30, LC1 comprises a sequence as shown in SEQ ID NO: 27 or SEQ ID NO: 31, and LC2 comprises a sequence as shown in SEQ ID NO: 28 or SEQ ID NO: 32; Preferably, HC1 comprises the sequence shown in SEQ ID NO: 25, HC2 comprises the sequence shown in SEQ ID NO: 26, LC1 comprises the sequence shown in SEQ ID NO: 27, and LC2 comprises the sequence shown in SEQ ID NO: 28, or The HC1 contains the sequence shown in SEQ ID NO: 29, the HC2 contains the sequence shown in SEQ ID NO: 30, the LC1 contains the sequence shown in SEQ ID NO: 31, and the LC2 contains the sequence shown in SEQ ID NO:

32.

4. The use according to any one of claims 1 to 3, wherein D is selected from the following compounds, and their pharmaceutically acceptable salts, solvates, tautomers, meso compounds, racemates, enantiomers, and diastereomers. Preferably, the D is linked to the -L- group in the antibody-drug conjugate via a hydroxyl, amino, primary, secondary, or tertiary amino group thereon.

5. The use according to any one of claims 1 to 4, wherein the structure of the anti-EGFR / c-Met antibody-drug conjugate and its tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates or isotope-labeled compounds is as shown in general formulas (2)-(12), in, BsAb is as defined in claims 1-3, -L- is as defined in claim 1, and y is selected from an integer or decimal of 1-12, preferably 2-8, more preferably 2-6 or 3-8, and even more preferably 2, 3, 4, 5, 6, or 7.

6. The use according to any one of claims 1 to 5, wherein the -L- is selected from -L1-L2-L3-L4-L5-, wherein, -L1- is a covalently connected unit, which is covalently connected to BsAb. -L2- is an extension unit. -L3- is selected from bonds or polar hydrophilic groups. -L4- is selected from peptide residues consisting of 1-8 amino acids. -L5- represents a bond, or a self-destructive fragment; Preferably, -L1- is selected from: -L2- is selected from: -(CH2)s 1 -C(=O)-,-CH2-C(=O)-NH-(CH2)s 1 -C(=O)-,-(CH2CH2O)s 2 -(CH2)s 1 -C(=O)-, Among them, S 1 and S 2 Each number is independently selected from 0-8; -L3- is selected from key, Among them, S 3 Selected from 2-12, preferably 6-12; -L4- is selected from peptide residues consisting of 1-8 amino acids, wherein the amino acids are selected from phenylalanine, isoleucine, leucine, tryptophan, valine, methionine, tyrosine, alanine, threonine, histidine, serine, glutamine, arginine, lysine, asparagine, glutamic acid, proline, citrulline, aspartic acid, and glycine; preferably, -L4- is selected from: -L5- is selected from: bond, -NH-CH2-, or 7. The use according to claim 6, wherein -L- is selected from...

8. The use according to any one of claims 1 to 7, wherein the structure of the anti-EGFR / c-Met antibody-drug conjugate and its tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates or isotopic labels is as shown in general formulas (13)-(24), in, D is selected from the following compounds and their pharmaceutically acceptable salts, solvates, tautomers, meso compounds, racemates, enantiomers, and diastereomers: Wherein, BsAb is defined as in claims 1-3, y is selected from an integer or decimal of 1-12, preferably 2-8, more preferably 2-6 or 3-8, and even more preferably 2, 3, 4, 5, 6, or 7.

9. The use according to any one of claims 1 to 8, wherein the structure of the anti-EGFR / c-Met antibody-drug conjugate and its tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates, or isotope-labeled compounds is selected from the following: in, BsAb is defined as in claims 1-3, where y is selected from an integer or decimal of 1-12, preferably 2-8, more preferably 2-6 or 3-8, and even more preferably 2, 3, 4, 5, 6, or 7.

10. The use according to any one of claims 1 to 9, wherein the single-dose dose of the anti-EGFR / c-Met antibody-drug conjugate is 1.0 mg / kg to 12.0 mg / kg, preferably 1.0 mg / kg, 2.0 mg / kg, 3.0 mg / kg, 4.0 mg / kg, 4.2 mg / kg, 4.5 mg / kg, 4.8 mg / kg, 5.0 mg / kg, 6.0 mg / kg, 7.0 mg / kg, 8.0 mg / kg, 9.0 mg / kg, 10.0 mg / kg, 11.0 mg / kg, or 12.0 mg / kg, more preferably 3.0 mg / kg, 4.0 mg / kg, 4.2 mg / kg, 4.5 mg / kg, 4.8 mg / kg, 5.0 mg / kg, 6.0 mg / kg, 7.0 mg / kg, 8.0 mg / kg, 9.0 mg / kg, or 10.0 mg / kg; The administration frequency is once a week, once every two weeks, once every three weeks, or once every four weeks, preferably once every two weeks or once every three weeks; Preferably, The single-dose administration of the anti-EGFR / c-Met antibody-drug conjugate is 3.0 mg / kg, 4.0 mg / kg, 4.2 mg / kg, 4.5 mg / kg, 4.8 mg / kg, 5.0 mg / kg, 6.0 mg / kg, 7.0 mg / kg, and 8.0 mg / kg, administered once every three weeks; or The single-dose doses of the anti-EGFR / c-Met antibody-drug conjugate are 3.0 mg / kg, 4.0 mg / kg, 4.2 mg / kg, 4.5 mg / kg, 4.8 mg / kg, 5.0 mg / kg, 6.0 mg / kg, 7.0 mg / kg, and 8.0 mg / kg, and the dosing frequency is once every two weeks.

11. The use according to any one of claims 1 to 10, wherein the cancer is lung cancer, head and neck cancer, colorectal cancer, nasopharyngeal cancer, esophageal cancer, esophagogastric junction cancer, oral cancer, oropharyngeal cancer, hypopharyngeal cancer, laryngeal cancer, tongue cancer, gastric cancer, liver cancer, kidney cancer, pancreatic cancer, spleen cancer, skin cancer, thymic cancer, thyroid cancer, prostate cancer, bladder cancer, vaginal cancer, testicular cancer, anal cancer, breast cancer, ovarian cancer, cervical cancer, epithelial cell carcinoma, or glioblastoma; preferably non-small cell lung cancer. (NSCLC), small cell lung cancer, large cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, head and neck squamous cell carcinoma, head and neck adenocarcinoma, colorectal adenocarcinoma, colorectal mucinous carcinoma, oral cavity, oropharynx, hypopharynx or larynx squamous cell carcinoma, nasopharyngeal carcinoma, esophageal adenocarcinoma, esophagogastric junction adenocarcinoma, esophagogastric junction squamous cell carcinoma; more preferably non-squamous NSCLC, squamous NSCLC, head and neck squamous cell carcinoma, colorectal adenocarcinoma, oral cavity, oropharynx, hypopharynx or larynx squamous cell carcinoma, nasopharyngeal carcinoma, esophageal squamous cell carcinoma, esophagogastric junction squamous cell carcinoma; Preferably, the cancer is a cancer expressing EGFR and / or c-Met, which is associated with wild-type EGFR, EGFR mutation, EGFR gene amplification, elevated levels of circulating HGF, wild-type c-Met, c-Met mutation, c-Met gene amplification, or mutant KRAS. Preferably, the EGFR mutation includes at least one mutation selected from the group consisting of: L718Q, G719A, G719X (X is any amino acid), E746K, L747S, E749Q, A750P, A755V, V765M, T790M, C797S, L858R, L858P, L861X (X is any amino acid) substitution; E746-A750 deletion; R748-P753 deletion; Ala(A) insertion between M766 and A767; Ser, Val, and Ala(SVA) insertion between S768 and V769; Asn and Ser(NS) insertion between P772 and H773. Insertion; insertion of one or more amino acids between D761 and E762, between A763 and Y764, between Y764 and Y765, between M766 and A767, between A767 and V768, between S768 and V769, between V769 and D770, between D770 and N771, between N771 and P772, between P772 and H773, between H773 and V774, and between V774 and C775; deletion of one or more of EGFR exon 19 and EGFR exon 20; insertion of one or more of EGFR exon 19, EGFR exon 20, S768I, L861Q, and G719X (where X is any amino acid); More preferably, the EGFR mutation includes exon 19 deletion and / or L858R mutation.

12. The use according to any one of claims 1 to 11, wherein the cancer is one that has failed, progressed, or is intolerable to previous treatment with one or more anticancer therapies.

13. The use according to claim 12, wherein the one or more anticancer therapies comprise one or more chemotherapeutic agents, checkpoint inhibitors, targeted anticancer therapies, or kinase inhibitors, or any combination thereof; Preferably, the one or more anticancer therapies include carboplatin, cisplatin, paclitaxel, gemcitabine, vinorelbine, docetaxel, PD-1 inhibitors, PD-L1 inhibitors, EGFR inhibitors, VEGFR inhibitors, c-Met inhibitors, HER2 inhibitors, HER3 inhibitors, HER4 inhibitors, or any combination thereof.

14. The use according to any one of claims 11 to 13, wherein the cancer is locally advanced or metastatic NSCLC, recurrent or metastatic head and neck squamous cell carcinoma, recurrent or metastatic colorectal adenocarcinoma, recurrent or metastatic oral, oropharyngeal, hypopharyngeal or laryngeal squamous cell carcinoma, metastatic nasopharyngeal carcinoma, recurrent or metastatic esophageal squamous cell carcinoma, or recurrent or metastatic esophagogastric junction squamous cell carcinoma. Preferred types include locally advanced or metastatic NSCLC carrying EGFR-sensitive mutations, metastatic NSCLC with EGFR wild-type, recurrent or metastatic head and neck squamous cell carcinoma that has failed or is intolerant to standard treatment, recurrent or metastatic colorectal adenocarcinoma that has failed or is intolerant to standard treatment, recurrent or metastatic squamous cell carcinoma of the oral cavity, oropharynx, hypopharynx, or larynx that has failed or is intolerant to standard treatment, metastatic nasopharyngeal carcinoma that has failed or is intolerant to adequate standard treatment, recurrent or metastatic esophageal squamous cell carcinoma that has failed or is intolerant to standard treatment, and recurrent or metastatic esophagogastric junction squamous cell carcinoma that has failed or is intolerant to standard treatment. More preferably, locally advanced or metastatic NSCLC with EGFR-sensitive mutations who have received systemic therapy with EGFR TKIs, metastatic NSCLC with EGFR wild-type who have previously received platinum-based chemotherapy or PD-1 / L1 inhibitor monotherapy or combination therapy, recurrent or metastatic head and neck squamous cell carcinoma who have previously received platinum-based chemotherapy or PD-1 / L1 inhibitor monotherapy or combination therapy, recurrent or metastatic colorectal adenocarcinoma that has failed or is intolerant to standard treatment and does not carry KRAS mutations, NRAS mutations or BRAF V600E mutations, recurrent or metastatic squamous cell carcinoma of the oral cavity, oropharynx, hypopharynx or larynx that have previously received platinum-based chemotherapy or PD-1 / L1 inhibitor monotherapy or combination therapy, and metastatic nasopharyngeal carcinoma that have previously received platinum-based chemotherapy or PD-1 / L1 inhibitor monotherapy or combination therapy; Preferably, the EGFR-sensitive mutations include EGFR exon 19 deletion and / or L858R mutation; the KRAS mutations include exon 2, 3, and 4 mutations; and the NRAS mutations include exon 2, 3, and 4 mutations.

15. A pharmaceutical composition comprising an anti-EGFR / c-Met antibody-drug conjugate as defined in any one of claims 1 to 10, and its tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates, or isotope labels.

16. A kit or article comprising an anti-EGFR / c-Met antibody-drug conjugate as defined in any one of claims 1 to 10, and its tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates, or isotopic labels.

17. Use of the pharmaceutical composition of claim 15, or the cassette or article of claim 16, in the preparation of a medicament for treating cancer; Preferably, the cancer is as defined in any one of claims 11 to 14.

18. A method for preventing and / or treating cancer, comprising administering to a subject in need an anti-EGFR / c-Met antibody-drug conjugate and its tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates or isotopic labels. The dosage of the anti-EGFR / c-Met antibody-drug conjugate and its tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates, or isotope-labeled compounds is 1.0 mg / kg to 12.0 mg / kg, preferably 1.0 mg / kg, 2.0 mg / kg, 3.0 mg / kg, 4.0 mg / kg, 4.2 mg / kg, 4.5 mg / kg, 4.8 mg / kg, and 5.0 mg / kg. 6.0mg / kg, 7.0mg / kg, 8.0mg / kg, 9.0mg / kg, 10.0mg / kg, 11.0mg / kg, 12.0mg / kg, more preferably 3.0mg / kg, 4.0mg / kg, 4.2mg / kg, 4.5mg / kg, 4.8mg / kg, 5.0mg / kg, 6.0mg / kg, 7.0mg / kg, 8.0mg / kg, 9.0mg / kg, 10.0mg / kg; The administration frequency is once a week, once every two weeks, once every three weeks, or once every four weeks, preferably once every two weeks or once every three weeks; Preferably, The single-dose dosage is 3.0 mg / kg, 4.0 mg / kg, 4.2 mg / kg, 4.5 mg / kg, 4.8 mg / kg, 5.0 mg / kg, 6.0 mg / kg, 7.0 mg / kg, and 8.0 mg / kg, and the dosing frequency is once every three weeks; or The single-dose dosages are 3.0 mg / kg, 4.0 mg / kg, 4.2 mg / kg, 4.5 mg / kg, 4.8 mg / kg, 5.0 mg / kg, 6.0 mg / kg, 7.0 mg / kg, and 8.0 mg / kg, and the dosing frequency is once every two weeks; Preferably, The anti-EGFR / c-Met antibody-drug conjugate and its tautomers, mesosomes, racemates, enantiomers, diastereomers, pharmaceutical salts, hydrates, solvates or isotope labels are as defined in any one of claims 1 to 9. The cancer is as defined in any one of claims 11 to 14.

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