Treatment of nasopharyngeal carcinoma with antibody-drug conjugates targeting EGFR

Abstract

The present invention relates to the treatment of nasopharyngeal carcinoma using an antibody-drug conjugate targeting EGFR. Specifically, the present invention has demonstrated that the antibody-drug conjugate MYK-3, administered at 2.3 mg / kg once every three weeks, can effectively treat cancers such as EGFR-positive, HER2-negative advanced gastric cancer, recurrent or metastatic nasopharyngeal carcinoma, recurrent or metastatic head and neck squamous cell carcinoma, EGFR-positive advanced biliary adenocarcinoma, and EGFR-positive advanced non-small cell lung cancer. Furthermore, clinical use of 2.3 mg / kg MYK-3 has been shown to produce better tumor suppression or delay effects in patients than a lower dose of 2.0 mg / kg, and adverse drug reactions can be safely controlled.

Description

[Technical Field]

[0001] The present invention relates to the field of medicinal chemistry, specifically to the use of antibody drug conjugates, and more specifically to the treatment of nasopharyngeal carcinoma with antibody drug conjugates that target EGFR. [Background technology]

[0002] The epidermal growth factor receptor (EGFR), also known as HER1 or c-ErbB1, is a cell surface receptor of the epidermal growth factor family. It is a transmembrane glycoprotein consisting of 1186 amino acid residues and a molecular weight of 170 kD. EGFR belongs to the type I tyrosine kinase receptor ErbB subfamily (ErbB1-4) and possesses tyrosine kinase activity. EGFR is stably expressed in many epithelial tissues, including skin and hair follicles. Abnormal expression of the epidermal growth factor receptor or activation by receptor mutations can lead to oncogenic transformation. Overexpression of the epidermal growth factor receptor has been found in many solid tumors, including colorectal, head and neck, lung, ovarian, cervical, bladder, and esophageal cancers. Growth factors such as transforming growth factor α and epidermal growth factor are endogenous ligands of the epidermal growth factor receptor. These ligands bind to the epidermal growth factor receptor, activate the receptor's intracellular tyrosine kinase, and initiate multiple downstream signaling pathways, thereby regulating the growth and differentiation of normal cells, increasing the invasiveness of tumor cells, promoting angiogenesis, and inhibiting apoptosis of tumor cells. Because the epidermal growth factor receptor is overexpressed in tumors and plays an important role in tumor cell proliferation and differentiation, it has attracted attention as a promising target for tumor therapy.

[0003] Antibody-drug conjugates (ADCs) are a new type of biopharmaceutical that combines a monoclonal antibody (mAb) targeting a specific antigen with a potent cytotoxic small molecule drug (payload) via a linker. ADCs utilize the specific recognition function of antibodies to guide small molecule drugs to target cancer cells, allowing them to be internalized within the cancer cells, and then release the cytotoxic drug, thereby specifically killing the cancer cells. Since its introduction, ADCs have undergone continuous innovation and optimization, and have achieved great success in tumor treatment.

[0004] Currently, anti-EGFR antibody-drug conjugates are being developed one after another, such as the anti-EGFR antibody-drug conjugate MYK-3 disclosed in registered patent CN106999606B. However, the clinical use of the anti-EGFR antibody-drug conjugate MYK-3 has not been fully studied, and further exploration and solutions are needed. Summary of the Invention

[0005] Through extensive experimentation and creative research, the inventors of this application have discovered that the antibody-drug conjugate MYK-3, when administered clinically at 2.3 mg / kg once every three weeks, is effective in treating cancers such as EGFR-positive, HER2-negative advanced gastric cancer, recurrent or metastatic nasopharyngeal cancer, recurrent or metastatic head and neck squamous cell carcinoma, EGFR-positive advanced biliary adenocarcinoma, and EGFR-positive advanced non-small cell lung cancer. Furthermore, compared to a lower dose of 2.0 mg / kg, clinical administration of 2.3 mg / kg of MYK-3 has been shown to be more effective in inhibiting tumors and delaying progression in patients, and adverse drug reactions are safely manageable.

[0006] Thus, in one aspect, the present invention provides an antibody-drug conjugate, or a pharmaceutically acceptable salt, solvate, or solvate of said salt, for treating an epidermal growth factor receptor (EGFR)-associated disease.

[0007] The antibody-drug conjugate has a structure represented by Formula I:

[0008] Ab-(LD) p Formula I where: Ab represents an anti-epidermal growth factor receptor antibody, which comprises a heavy chain and a light chain, wherein the heavy chain variable regions CDR1, CDR2, and CDR3 comprise the sequences shown in SEQ ID NO:5, SEQ ID NO:6, and SEQ ID NO:7, respectively, or variants thereof, and the light chain variable regions CDR1, CDR2, and CDR3 comprise the sequences shown in SEQ ID NO:12, SEQ ID NO:13, and SEQ ID NO:14, respectively, or variants thereof.

[0009] L represents a linker, and the linker is 6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (MC-vc-PAB).

[0010] D represents a cytotoxic agent, said cytotoxic agent being MMAE.

[0011] p is any number selected from 1 to 8 (e.g., 1, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5 .7, 5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9 or 8, or 1-1.5, 1.5-2, 2-2.5, 2.5-3, 3-3.5, 3.5-4, 4-4.5, 4.5-5, 5-5.5, 5.5-6, 6-6.5, 6.5-7, 7-7.5 or 7.5-8); Here, the antibody-drug conjugate, or a pharmaceutically acceptable salt, solvate, or solvate of the salt thereof is administered in a total dose of 2.1 to 2.4 mg / kg every three weeks.

[0012] In another aspect, the present invention provides a method for treating a disease associated with epidermal growth factor receptor (EGFR), comprising administering to a subject an antibody-drug conjugate, or a pharmaceutically acceptable salt, solvate, or solvate of said salt thereof.

[0013] The antibody-drug conjugate has the structure shown in Formula I:

[0014] Ab-(LD) p Formula I where: Ab represents an anti-epidermal growth factor receptor antibody, which comprises a heavy chain and a light chain, wherein the heavy chain variable regions CDR1, CDR2, and CDR3 comprise the sequences shown in SEQ ID NO:5, SEQ ID NO:6, and SEQ ID NO:7, respectively, or variants thereof, and the light chain variable regions CDR1, CDR2, and CDR3 comprise the sequences shown in SEQ ID NO:12, SEQ ID NO:13, and SEQ ID NO:14, respectively, or variants thereof.

[0015] L represents a linker, and the linker is 6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (MC-vc-PAB).

[0016] D represents a cytotoxic agent, said cytotoxic agent being MMAE.

[0017] p is any number selected from 1 to 8 (e.g., 1, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5 .7, 5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9 or 8, or 1-1.5, 1.5-2, 2-2.5, 2.5-3, 3-3.5, 3.5-4, 4-4.5, 4.5-5, 5-5.5, 5.5-6, 6-6.5, 6.5-7, 7-7.5 or 7.5-8).

[0018] Here, the antibody-drug conjugate, or a pharmaceutically acceptable salt, solvate, or solvate of the salt thereof is administered in a total dose of 2.1 to 2.4 mg / kg every three weeks.

[0019] In Formula I, LD represents a covalent bond between the linker and the cytotoxic agent to form an LD molecule, Ab-(LD) p indicates that p LD molecules are covalently bound to Ab.

[0020] In the present invention, the drug-antibody ratio (DAR) refers to the number of drug molecules (also called cytotoxic agents) conjugated to an antibody (e.g., p in Formula I). ​​The number of drug molecules contained in the antibody-drug conjugates described herein may be an integer or a decimal. Regardless of whether it is an integer or a decimal, it refers to the average number of drug molecules conjugated per antibody. "p is an arbitrary number selected from 1 to 8" means that p may be any integer selected from 1 to 8 (including both the 1 and 8 ends) or any decimal number selected from 1 to 8, such as 4.1. At the same time, those skilled in the art will understand that even when using the same preparation method, the DAR values ​​of antibody-drug conjugates prepared in different batches are not necessarily completely identical and may vary, for example, within a range not exceeding 0.5.

[0021] The drug-antibody ratio (DAR) can be measured by conventional means, such as mass spectrometry, ELISA assay, HIC, and HPLC. The quantitative distribution of ADCs with respect to p can also be measured. In some cases, separation, purification, and verification of homogeneous ADCs with a certain value of p from ADCs with other drug loads can be achieved by means such as HIC, reverse-phase HPLC, or electrophoresis.

[0022] Here, the structural formula of MC-vc-PAB is as follows: [ka] The structural formula of MMAE is as follows: [ka] In Formula I, LD is MC-vc-PAB-MMAE, the structural formula of which is as follows: [ka] When the above LD, i.e., MC-vc-PAB-MMAE, is covalently bound to Ab, it is formed by coupling between the succinimide at the end of the LD and the thiol group in the antibody, and the structural formula of the formed ADC is as follows:

[0023] [ka]

[0024] However, in the ADC formed above, the antibody Ab is bound to the carbon atom of the succinimide at the LD end via -S-, and this -S- is not a thiol group of the Ab introduced separately, but is a thiol group contained in the antibody itself after the antibody Ab is reduced to open the disulfide bond.

[0025] In some embodiments, the antibody-drug conjugate, or a pharmaceutically acceptable salt, solvate, or solvate of said salt thereof is administered at a total dose of 2.2 to 2.4 mg / kg every three weeks.

[0026] In some embodiments, the antibody-drug conjugate, or a pharmaceutically acceptable salt, solvate, or solvate of said salt thereof, is administered at a total dose of 2.3 mg / kg every three weeks.

[0027] The term "every 3 weeks" refers to a treatment cycle of 3 weeks, and the term "total dose every 3 weeks" refers to the total dose within that treatment cycle (i.e., every 3 weeks). For example, if the total dose every 3 weeks is 2.3 mg / kg, the administration regimen may be a single high dose but infrequent administration (e.g., a single dose of 2.3 mg / kg, but once every 3 weeks), or a single low dose but frequent administration (e.g., a single dose of 0.77 mg / kg, but once every 7 days, i.e., once a week). The specific number of treatment cycles may be, for example, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32, at least 33, at least 34, at least 35, at least 36, at least 37, at least 38, at least 39, or at least 40, or even more. In some embodiments, the specific setting or selection of the number of treatment cycles is based on the principle that treatment will continue until disease progression is confirmed. Additionally, one skilled in the art should understand that the dosage may vary slightly above and below that amount by up to 0.05 mg / kg.

[0028] In some embodiments, the single dose of the antibody-drug conjugate, or a pharmaceutically acceptable salt, solvate, or solvate of the salt thereof is 2.1 to 2.4 mg / kg (preferably 2.2 to 2.4 mg / kg, more preferably 2.3 mg / kg), and the administration frequency is once every three weeks. In some embodiments, the administration is on the first day of every three weeks.

[0029] In some embodiments, the single dose of the antibody-drug conjugate, or a pharmaceutically acceptable salt, solvate, or solvate of the salt thereof is 0.7 to 1.4 mg / kg (e.g., 0.7 to 1.1 mg / kg, specifically 0.77 mg / kg or 1.1 mg / kg), and the administration frequency is once every 7 to 10 days.

[0030] In some embodiments, the method of administration of the antibody-drug conjugate, or a pharmaceutically acceptable salt, solvate, or solvate of said salt thereof is intravenous infusion (also called intravenous administration, intravenous injection, etc.).

[0031] In some embodiments, the subject to which the antibody-drug conjugate, or a pharmaceutically acceptable salt, solvate, or solvate of said salt thereof is administered is a human.

[0032] In some embodiments, the subject to which the antibody-drug conjugate, or a pharmaceutically acceptable salt, solvate, or solvate of said salt thereof is administered is a patient with a disease associated with epidermal growth factor receptor (EGFR).

[0033] In some embodiments, the antibody-drug conjugate, or a pharmaceutically acceptable salt, solvate, or solvate of the salt thereof, is administered to a patient who has previously been treated with an anti-EGFR drug, preferably an anti-EGFR antibody.

[0034] In some embodiments, the subject to which the antibody-drug conjugate, or a pharmaceutically acceptable salt, solvate, or solvate of said salt thereof is administered is a patient who overexpresses EGFR.

[0035] In some embodiments, the antibody-drug conjugate, or a pharmaceutically acceptable salt, solvate, or solvate of said salt thereof, is administered to patients with recurrent or metastatic nasopharyngeal carcinoma (NPC) who have previously received systemic treatment with at least a first-line platinum-containing therapy and a PD-1 / PD-L1 inhibitor and whose disease has progressed during treatment, recurred after treatment, or was intolerant to treatment.

[0036] In some embodiments, the antibody-drug conjugate, or a pharmaceutically acceptable salt, solvate, or solvate of said salt thereof, is administered to patients with recurrent or metastatic nasopharyngeal carcinoma (NPC) who have previously received at least second-line systemic chemotherapy and PD-1 / PD-L1 therapy but have failed. In some embodiments, if the patient has previously received at least second-line systemic chemotherapy, the chemotherapy regimen must include at least a platinum-containing therapy, gemcitabine, and a taxane / capecitabine.

[0037] In some embodiments, the subject is a human.

[0038] In some embodiments, the subject is a patient with an epidermal growth factor receptor (EGFR)-associated disease.

[0039] In some embodiments, the subject has previously been treated with an anti-EGFR drug, preferably an anti-EGFR antibody.

[0040] In some embodiments, the subject is an EGFR-overexpressing patient.

[0041] In some embodiments, the subject has recurrent or metastatic nasopharyngeal carcinoma (NPC) who has previously been systematically treated with at least a first-line platinum-containing therapy and a PD-1 / PD-L1 inhibitor and whose disease has progressed during treatment, recurred after treatment, or was intolerant to treatment.

[0042] In some embodiments, the subject has recurrent or metastatic nasopharyngeal carcinoma (NPC) who has previously received at least second-line systemic chemotherapy and PD-1 / PD-L1 therapy but failed the regimen. In some embodiments, the chemotherapy regimen must include at least a platinum-containing therapy, gemcitabine, and a taxane / capecitabine.

[0043] In some embodiments, the taxane drugs include paclitaxel, docetaxel, albumin-bound paclitaxel, and paclitaxel liposomes.

[0044] In some embodiments, the platinum-containing therapy includes cisplatin, carboplatin, nedaplatin, or the like.

[0045] In some embodiments, the platinum-containing therapy includes, but is not limited to, the following therapies: Cisplatin plus gemcitabine (first-line); or Platinum (such as cisplatin or carboplatin) plus taxane (such as docetaxel or paclitaxel); or Platinum + EGFR monoclonal antibody (e.g., cetuximab or nimotuzumab).

[0046] In some embodiments, the epidermal growth factor receptor (EGFR)-associated disease is cancer.

[0047] In some embodiments, the cancer is selected from nasopharyngeal cancer, head and neck squamous cell carcinoma, biliary tract cancer, esophageal cancer, duodenal cancer, colorectal cancer, colon cancer, rectal cancer, head and neck cancer, lung cancer (e.g., non-small cell lung cancer), ovarian cancer, cervical cancer, bladder cancer, esophageal cancer, breast cancer, kidney cancer, prostate cancer, gastric cancer, pancreatic cancer, and glioma.

[0048] In some embodiments, the cancer is selected from nasopharyngeal carcinoma, head and neck squamous cell carcinoma, non-small cell lung cancer, biliary tract cancer, gastric cancer, esophageal cancer, and duodenal cancer.

[0049] In some embodiments, the cancer is nasopharyngeal carcinoma.

[0050] In some embodiments, the cancer is an advanced stage, heavily pre-treated solid tumor.

[0051] In some embodiments, the cancer is a recurrent or metastatic EGFR-positive advanced solid tumor after multiple prior tumor-related treatments.

[0052] In some embodiments, the cancer is selected from EGFR-positive, HER2-negative advanced gastric cancer, recurrent or metastatic nasopharyngeal carcinoma, recurrent or metastatic head and neck squamous cell carcinoma, EGFR-positive advanced biliary adenocarcinoma, EGFR-positive advanced non-small cell lung cancer, etc. Here, "EGFR-positive" refers to overexpression of EGFR, and "HER2-negative" refers to the absence of overexpression of HER2.

[0053] In some embodiments, the cancer is recurrent metastatic nasopharyngeal carcinoma.

[0054] In some embodiments, p is any number selected from 2-6.

[0055] In some embodiments, p is any number selected from 3-5.

[0056] In some embodiments, the FR1, FR2, FR3, and FR4 regions of the heavy chain variable region of the anti-epidermal growth factor receptor antibody comprise the sequences set forth in SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, or variants thereof, respectively.

[0057] In some embodiments, the FR1, FR2, FR3, and FR4 regions of the light chain variable region of the anti-epidermal growth factor receptor antibody comprise the sequences set forth in SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, or variants thereof, respectively.

[0058] In some embodiments, the heavy chain constant region of the anti-epidermal growth factor receptor antibody is selected from human IgG, IgM, IgA, IgD, IgE constant regions or variants of these constant regions.

[0059] In some embodiments, the IgG is selected from IgG1, IgG2, IgG3, and IgG4.

[0060] In some embodiments, the light chain constant region of the anti-epidermal growth factor receptor antibody is a human lambda constant region, a kappa constant region, or a variant of these constant regions.

[0061] In some embodiments, the sequence of the heavy chain variable region of the anti-epidermal growth factor receptor antibody comprises the sequence set forth in SEQ ID NO:1 or a sequence having greater than 70%, preferably greater than 75%, 80%, 85%, 90%, 95%, or 99% identity to the sequence set forth in SEQ ID NO:1.

[0062] In some embodiments, the sequence of the heavy chain variable region of the anti-epidermal growth factor receptor antibody is set forth in SEQ ID NO:1.

[0063] In some embodiments, the sequence of the light chain variable region of the anti-epidermal growth factor receptor antibody comprises the sequence set forth in SEQ ID NO:2, or a sequence having greater than 70%, preferably greater than 75%, 80%, 85%, 90%, 95%, or 99% identity to the sequence set forth in SEQ ID NO:2.

[0064] In some embodiments, the sequence of the light chain variable region of the anti-epidermal growth factor receptor antibody is set forth in SEQ ID NO:2.

[0065] In some embodiments, the sequence of the heavy chain constant region of the anti-epidermal growth factor receptor antibody comprises the sequence set forth in SEQ ID NO: 3 or a sequence having greater than 70%, preferably greater than 75%, 80%, 85%, 90%, 95%, or 99% identity to the sequence set forth in SEQ ID NO: 3.

[0066] In some embodiments, the sequence of the heavy chain constant region of the anti-epidermal growth factor receptor antibody is that shown in SEQ ID NO:3.

[0067] In some embodiments, the sequence of the light chain constant region of the anti-epidermal growth factor receptor antibody comprises the sequence set forth in SEQ ID NO: 4 or a sequence having greater than 70%, preferably greater than 75%, 80%, 85%, 90%, 95%, or 99% identity to the sequence set forth in SEQ ID NO: 4.

[0068] In some embodiments, the sequence of the light chain constant region of the anti-epidermal growth factor receptor antibody is set forth in SEQ ID NO:4.

[0069] Beneficial effects The objective of the present invention is to determine the clinical dosage and indications of MYK-3. Based on Phase I / II clinical studies, the inventors have found that MYK-3 administered at 2.3 mg / kg once every three weeks can treat EGFR-positive, HER2-negative advanced gastric cancer, recurrent metastatic nasopharyngeal carcinoma, recurrent or metastatic head and neck squamous cell carcinoma, EGFR-positive advanced biliary adenocarcinoma, and EGFR-positive advanced non-small cell lung cancer. MYK-3 exhibits better therapeutic effects in the expanded spectrum of recurrent metastatic nasopharyngeal carcinoma. Compared with doses of 2.0 mg / kg and 2.5 mg / kg, the inventors have found that clinical use of 2.3 mg / kg MYK-3 resulted in better tumor inhibition or delay in patients, and adverse drug reactions were safely controlled. [Brief explanation of the drawings]

[0070] [Figure 1] FIG. 1 shows an HIC-HPLC diagram measuring the drug / antibody ratio of an antibody-drug conjugate.

[0071] [Figure 2] FIG. 2 shows in vitro cell activity assays of monoclonal antibodies and antibody-drug conjugates, where ○ indicates BA03 monoclonal antibody and ▲ indicates MYK-3 antibody-drug conjugate.

[0072] [Figure 3]FIG. 3 shows the growth inhibitory activity of MYK-3 against colon cancer cell HT-29, where ○ indicates the BA03 monoclonal antibody and ▲ indicates the MYK-3 antibody-drug conjugate.

[0073] [Figure 4] FIG. 4 shows the growth inhibitory activity of MYK-3 against glioma cells U87-MG, where ○ indicates the BA03 monoclonal antibody and ▲ indicates the MYK-3 antibody-drug conjugate.

[0074] [Figure 5] Figure 5 shows the growth inhibitory activity of MYK-3 against lung cancer cell A549, where ○ indicates the BA03 monoclonal antibody and ▲ indicates the MYK-3 antibody-drug conjugate.

[0075] [Figure 6] Figure 6 shows the growth inhibitory activity of MYK-3 against KRAS mutant colon cancer cells LoVo, where ◇ indicates the cetuximab monoclonal antibody and ▲ indicates the MYK-3 antibody-drug conjugate. DETAILED DESCRIPTION OF THE INVENTION

[0076] Hereinafter, embodiments of the present invention will be described in detail with reference to examples. However, it should be understood by those skilled in the art that these examples are for the purpose of illustrating the present invention and do not limit the scope of the present invention. Examples for which specific conditions are not specified were carried out under standard conditions or conditions recommended by the manufacturer. Reagents and equipment used were not specified by manufacturers and were all commercially available and standard.

[0077] In the present invention, unless otherwise specified, scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. In addition, the terms related to protein and nucleic acid chemistry, molecular biology, cell and tissue culture, microbiology, immunology, and laboratory procedures used herein are terms and routine procedures widely used in the respective fields. Meanwhile, to facilitate understanding of the present invention, definitions and explanations of relevant terms are provided below.

[0078] In the present invention, unless otherwise specified, any numerical range is intended to include any value or any subrange within that range.

[0079] In the present invention, the term "antibody" refers to an immunoglobulin molecule that typically consists of two pairs of identical polypeptide chains, each pair having one "light" (L) chain and one "heavy" (H) chain. Antibody light chains can be classified into two types: kappa and lambda. Heavy chains can be classified into five types: μ, delta, gamma, alpha, or epsilon, and heavy chains can be classified into five types: IgM, IgD, IgG, IgA, and IgE. Within the light and heavy chains, the variable and constant regions are connected by a "J" region of about 12 or more amino acids, and heavy chains also contain a "D" region of about 3 or more amino acids. Each heavy chain contains a heavy chain variable region (V H ) and heavy chain constant region ( C H The heavy chain constant region consists of three domains (C H 1. C H 2, and C H Each light chain consists of a light chain variable region (V L ) and light chain constant region ( C L The light chain constant region consists of one domain, C L The constant regions of the antibody may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the complement system component C1q. H and V L The region may also be subdivided into regions of high variability (called complementarity determining regions (CDRs)), interspersed with relatively conserved regions (called framework regions (FRs)). H and V L The variable region (V) of each heavy / light chain pair consists of three CDRs and four FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. H and V L) each form an antibody binding site. The assignment of amino acids to each region or domain follows the definitions of the Kabat Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987 and 1991)) or Chothia & Lesk (1987) J. Mol. Biol. 196:901-917; Chothia et al. (1989) Nature 342:878-883.

[0080] The antibody of the present invention can be prepared by various methods known in the art, for example, by recombinant genetic engineering techniques. For example, DNA molecules encoding the heavy and light chain genes of the antibody of the present invention can be obtained by chemical synthesis or PCR amplification. The obtained DNA molecules are inserted into an expression vector and then transfected into host cells. The transfected host cells are then cultured under specific conditions to express the antibody of the present invention.

[0081] In the present invention, algorithms used to determine sequence homology and sequence similarity include, for example, the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al. (1977) Nucl. Acid. Res. 25:3389-3402 and Altschul et al. (1990) J. Mol. Biol. 215:403-410, respectively. BLAST and BLAST 2.0 can be used to determine percent amino acid sequence homology in the present invention, for example, using parameters described in the literature or default parameters. Software for performing BLAST analysis is publicly available through the National Center for Biotechnology Information (NCBI).

[0082] In the present invention, an amino acid sequence having at least 70% sequence identity to the amino acid sequence includes a polypeptide sequence substantially identical to the amino acid sequence, for example, a sequence having at least 70%, preferably 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to a polypeptide sequence of the present invention, using methods described herein (e.g., BLAST analysis using standard parameters).

[0083] In the present invention, the term "variant of the amino acid sequence" refers to a sequence having more than 70% homology to the amino acid sequence, for example, more than 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% homology, including, for example, a sequence having 3, 2, or 1 substituted, deleted, or added amino acids. Preferably, the number of substituted, added, or deleted amino acids is 3 or less. More preferably, the number of substituted, added, or deleted amino acids is 2 or less. Most preferably, the number of substituted, added, or deleted amino acids is 1 or less.

[0084] A "substitutional" variant refers to a variant in which at least one amino acid residue in a native sequence has been removed and a different amino acid inserted in its place. The substitutions may be single, substituting only one amino acid in the molecule, or multiple, substituting two or more amino acids in the molecule. The substitutions may be in consecutive positions. Similarly, an amino acid may be replaced by multiple amino acid residues, in which case the variant encompasses both "substitution" and "insertion." An "insertional" (or "additional") variant refers to a variant in which one or more amino acids are inserted adjacent to a particular position in the native sequence, where "adjacent to an amino acid" refers to attachment to the α-carboxyl or α-amino functionality of that amino acid. A "deletional" variant refers to a variant in which one or more amino acids have been removed from the native amino acid sequence. Generally, deletional variants result in the deletion of one or two amino acids in a particular region of the molecule.

[0085] In the present invention, EGFR overexpression refers to an increase in EGFR expression levels compared to the EGFR expression level on the surface of normal epithelial cells. Specifically, EGFR expression can be classified into high, moderate, and low expression. For example, DiFi cells are a cell line with high EGFR expression, LoVo cells are a cell line with moderate EGFR expression, and HT-29 cells are a cell line with low EGFR expression (Wild, R., et al., Mol. Cancer Ther. 2006: 5(1), pp. 104-113; Cetuximab preclinical antitumor activity (monotherapy and combination-based) is not predicted by relative total or activated epidermal growth factor receptor tumor expression levels).

[0086] In the present invention, the KRAS gene is synonymous with the K-RAS gene, a member of the RAS gene family that encodes the K-ras protein and is involved in the development, proliferation, metastasis, spread, and angiogenesis of various tumors. Common mutation sites are codons 12 and 13 in exon 2 and codon 61 in exon 3 of the K-RAS gene, among which there are seven mutation hotspots, namely, G12C, G12R, G12S, G12V, G12D, G12A, and G13V / D, and these seven mutations account for more than 90% of tumors. In one embodiment of the present invention, the tumor is a KRAS gene mutation tumor associated with EGFR overexpression.

[0087] In the present invention, the 20 common amino acids and their abbreviations follow common usage. For details, please refer to Immunology - A Synthesis (2nd ed., ES Golub and DR Gren, eds., Sinauer Associates, Sunderland, Mass. (1991)). The above document is incorporated herein by reference.

[0088] In the present invention, less than the theoretical maximum number of drug moieties are coupled to an antibody in the coupling reaction. Generally, antibodies do not contain many free and reactive cysteine ​​thiol groups that can be conjugated to drug moieties. In practice, most of the cysteine ​​thiol groups in antibodies exist as disulfide bridges. In some embodiments, antibodies can be reduced under partial or complete reduction conditions using a reducing agent such as dithiothreitol (DTT) or tricarbonylethylphosphine (TCEP) to generate reactive cysteine ​​thiol groups.

[0089] In the present invention, the term "pharmaceutically acceptable salt" refers to (i) a salt formed from an acidic functional group present in the complex provided by the present invention and a suitable inorganic or organic cation (base), including, but not limited to, alkali metal salts such as sodium salt, potassium salt, and lithium salt; alkaline earth metal salts such as calcium salt and magnesium salt; other metal salts such as aluminum salt, iron salt, zinc salt, copper salt, nickel salt, and cobalt salt; inorganic base salts such as ammonium salt; and organic base salts such as tertiary octylamine salt, dibenzylamine salt, morpholine salt, glucosamine salt, phenylglycine alkyl ester salt, ethylenediamine salt, N-methylglucamine salt, guanidine salt, diethylamine salt, triethylamine salt, dicyclohexylamine salt, N,N'-dibenzylethylenediamine salt, chloroprocaine salt, procaine salt, diethanolamine salt, N-benzyl-phenethylamine salt, piperazine salt, tetramethylamine salt, and tris(hydroxymethyl)aminomethane salt. and (ii) salts formed from the basic functional groups present in the complexes provided by the present invention and suitable inorganic or organic anions (acids), including, but not limited to, hydrohalides such as hydrofluoride, hydrochloride, hydrobromide, and hydroiodide; inorganic acid salts such as nitrate, perchlorate, sulfate, and phosphate; lower alkanesulfonates such as methanesulfonate, trifluoromethanesulfonate, and ethanesulfonate; arylsulfonates such as benzenesulfonate and p-benzenesulfonate; organic acid salts such as acetate, malate, fumarate, succinate, citrate, tartrate, oxalate, and maleate; and amino acid salts such as glycine, trimethylglycine, arginine, ornithine, glutamate, and aspartate.

[0090] Pharmaceutically acceptable salts can be obtained using standard procedures well known in the art, for example, by reacting a sufficient amount of a basic material with a suitable acid to provide a pharmaceutically acceptable anion, or by reacting a sufficient amount of an acidic material with a suitable base to provide a pharmaceutically acceptable cation.

[0091] In the present invention, a solvate refers to a complex in solid or liquid form formed by coordination of the antibody-drug conjugate with solvent molecules. A hydrate is a specific form of a solvate and has coordinated water molecules. In the present invention, a hydrate is a preferred solvate.

[0092] In the present invention, the term "treatment" generally means obtaining a desired pharmacological and / or physiological effect. The effect may be preventative, by complete or partial prevention of a disease or its symptoms, and / or therapeutic, by partial or complete stabilization or cure of the disease and / or its side effects. As used herein, "treatment" covers any treatment for a patient disease, including (a) preventing a disease or condition occurring in a patient who is susceptible to the disease or condition but has not yet been diagnosed with the disease, (b) suppressing the symptoms of the disease, i.e., preventing the progression of the disease, or (c) alleviating the symptoms of the disease, i.e., causing regression of the disease or condition.

[0093] In the present invention, the term "subject" refers to a vertebrate. In some embodiments, a vertebrate refers to a mammal. Mammals include, but are not limited to, livestock (such as cows), pets (such as cats, dogs, and horses), primates, mice, and rats. In some embodiments, a mammal refers to a human.

[0094] The antibody BA03 of the present invention is BA03 of Chinese patent application CN103772504A, and its preparation method is described in Example 3 of the same patent application. The sequences of each part of the antibody are as follows:

[0095] Heavy chain variable region sequence: QVQLQESGPGLVKPSETLSLTTCTVSGFSLS NYDVH WVRQAPGKGLEWLG VIWSGGNTDYNTPFTS RLTISVDTSKNQFSLKLSSVTAADTAVYYCAR ALDYYDYEFAY WGQGTLVTVSS (SEQ ID NO: 1).

[0096] Here, the underlined parts are CDR1 (SEQ ID NO: 5), CDR2 (SEQ ID NO: 6), and CDR3 (SEQ ID NO: 7), respectively.

[0097] The non-underlined portions are FR1 (SEQ ID NO:8), FR2 (SEQ ID NO:9), FR3 (SEQ ID NO:10), and FR4 (SEQ ID NO:11), respectively. Light chain variable region sequence: EIVLTQSPDFQSVTPKEKVTITC RASQSIGTNIH WYQQKPDQSPKLLIK YASESIS GIPSRFSGSGSGTDFTLTINSLEAEDAATYYC QQNNEWPTSF GQGTKLEIK (SEQ ID NO: 2).

[0098] Here, the underlined parts are CDR1 (SEQ ID NO: 12), CDR2 (SEQ ID NO: 13), and CDR3 (SEQ ID NO: 14), respectively.

[0099] The non-underlined portions are FR1 (SEQ ID NO:15), FR2 (SEQ ID NO:16), FR3 (SEQ ID NO:17), and FR4 (SEQ ID NO:18), respectively. Heavy chain constant region sequence: ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQ ID NO:3). Light chain constant region sequence: RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO:4). The present invention will be further described below using specific examples, but these examples should not be construed as limiting the scope of the present invention.

[0100] Example 1: Method for producing antibody-drug conjugates Ten milligrams of BA03 antibody was taken and substituted three times with reducing buffer (25 mM sodium borate, pH 8.0, 25 mM NaCl, 5 mM EDTA) using a 15 mL 30 KD ultrafiltration device. The substituted solution was transferred to a new Eppendorf tube (weighed), weighed, and the protein concentration was measured to calculate the total protein content. 2.5 times the molar equivalent of DTT was added to the antibody, and the mixture was incubated at room temperature for 2 hours. After continuous mixing, the substituted solution was substituted three times with coupling buffer (50 mM Tris, pH 7.2, 150 mM NaCl, 5 mM EDTA) using a 15 mL 30 KD ultrafiltration device. The concentrate was sampled, the protein concentration measured at A280, and the weight was used to calculate the total protein content. A 10 μL sample was then taken and the number of free thiol groups was measured using Ellman's method.

[0101] The molar concentration of free thiol was calculated using the formula shown below.

[0102] [ka] b: Optical path length of the colorimetric plate (usually 1 cm).

[0103] The number of moles of free thiol was calculated based on the molar concentration of free thiol and the volume of the total protein solution.

[0104] To the reduced antibody, vc-MMAE (Shanghai Haoyuan Chemexpress Co., Ltd., product number HY-15575) (dissolved in DMSO) was added in an amount equivalent to 1.1 times the molar amount of free thiols. The mixture was then mixed and allowed to react at room temperature for 2 hours with intermittent mixing. N-acetylcysteine ​​was added in an amount equivalent to 20 times the molar amount of vc-MMAE added to the reaction mixture, mixed, and allowed to stand for 5 minutes. The mixture was then substituted three times with conjugate stock (20 mM sodium citrate, 0.3% NaCl, 5% trehalose, 0.05% Tween-80, pH 6.0) using a 15 mL 30 kD ultrafiltration device to obtain the antibody-drug conjugate MYK-3. The sample was stored at 4°C.

[0105] Drug / Antibody Ratio Measurement: The prepared antibody-drug conjugate MYK-3 was analyzed by HIC-HPLC (Jun Ouyang, Drug-To-Antibody (DAR) Ratio and Drug Distribution by Hydrophobic Interaction Chromatography and Reverse Phase High Performance Chromatography, Laurent Ducry (ed.), Antibody Drug Conjugates, Chapter 17, Methods in Molecular Biology, Vol. 1045, pp. 275-283) to measure the drug-to-antibody ratio (DAR). As shown in Figure 1, the average drug loading number DAR calculated from the spectral peak area was 4.1.

[0106] Example 2: Assay of in vitro cellular activity of antibody drug conjugate MYK-3 Methods for detecting cell activity: After 3-4 passages, the restored cell lines were first removed from the medium, rinsed once with 5 mL of DPBS, digested with 3 mL of trypsin, resuspended in medium, centrifuged, and the supernatant discarded. The cells were then resuspended in medium again, and 0.5 mL of the supernatant was counted using a cell counter. The cells were seeded into 96-well cell culture plates (DiFi cells at 10,000 cells / well, HT-29 cells at 5,000 cells / well, A549 cells at 2,000 cells / well, U87-MG cells at 3,000 cells / well, and LoVo cells at 4,000 cells / well) and incubated for 24 hours. After incubation, serial dilutions of the monoclonal antibody BA03 and the antibody-drug conjugate MYK-3 were added and incubated for 72 hours. Then, 20 μL of CCK8 color reagent was added to each well, and OD450-650 was measured at wavelengths of 450-650 nm using a microplate reader, followed by four-parameter fitting.

[0107] In vitro cell activity experiment results: The following cell lines were purchased from the Shanghai Institute of Cell Biology, Chinese Academy of Sciences.

[0108] Activity in DiFi cells (human colorectal cancer cells) where EGFR is highly expressed: As shown in Figure 2, MYK-3 showed significantly increased cytostatic activity compared to monoclonal antibody BA03, and EC 50 was reduced by approximately 10 times (EC 50 was 51.9ng / mL, and the EC 50 is 5.1ng / mL).

[0109] Activity in other tumor cells with moderate or low EGFR expression: Compared to the monoclonal antibody itself, MYK-3 also exhibited significant cell growth inhibitory activity against cancer cells with moderate or low EGFR expression (human colon cancer cell HT-29, human lung cancer cell A549, and human astroblastoma cell U87-MG) (see Figures 3, 4, and 5). 50 EC of 611 ng / mL, A549 50 EC of U87-MG was 28.3 μg / mL. 50 was 5.3 μg / mL.

[0110] Furthermore, we also evaluated the activity of MYK-3 in LoVo, a KRAS mutant colon cancer cell line that overexpresses EGFR (Dunn EF, Ilda M, Myers RA, Hintz KA, Campbell DA, Armstrong EA, Li C and Wheeler DL. Dasatinib sensitizes KRAS mutant colorectal tumors to cetuximab. Oncogene 2011; 30:561-574). MYK-3 exhibited significant tumor growth suppression activity against LoVo KRAS mutant colon cancer cells (as shown in Figure 6). 50 = 3.2 μg / mL), whereas BA03 alone showed almost no inhibitory activity against the cell line.

[0111] Example 3: Clinical Trial of Antibody Drug Conjugate MYK-3 The study results are based on results from a Phase I clinical trial and an ongoing Phase II extension study.

[0112] 1) Clinical phase I Research method: In a Phase 1a dose-escalation study with a 3+3 design, patients with locally advanced or metastatic solid tumors who were refractory to or unable to undergo standard therapy received MYK-3 every 3 weeks (Q3W) for up to eight cycles. The starting dose of MYK-3 was 0.1 mg / kg, followed by dose escalation to 0.3, 0.6, 1.0, 1.5, 2.0, 2.5, and 3.0 mg / kg. The Phase 1b study included expansion cohorts for patients with advanced or metastatic colorectal cancer (CRC), squamous cell carcinoma of the head and neck (SCCHN), and nasopharyngeal carcinoma (NPC), who had progressed on multiple prior therapies and received MYK-3 at 2.5 mg / kg Q3W. Outcomes included adverse events (AEs), dose-limiting toxicities (DLTs), and antitumor activity assessed every two cycles.

[0113] result: A total of 22 patients without an EGFR-positive prescreen were enrolled in the phase Ia clinical trial. A total of 39 patients with CRC (12 patients), SCCHN (13 patients), and NPC (14 patients) who passed the EGFR-IHC prescreen were enrolled in the phase Ib clinical trial. The median ages of patients in phase Ia and Ib were 54.5 and 50.4 years, respectively, and the median number of prior therapies was 3 (1, 8) and 2 (1, 6), respectively. The dose in phase Ib was 2.5 mg / kg. In Phase I, common treatment-related adverse events (TRAEs) occurring in ≥20% of patients were rash, elevated aspartate aminotransferase (39%), alopecia (33%), decreased appetite (31%), elevated alanine aminotransferase (30%), pruritus (28%), decreased white blood cell count (25%), myalgia (23%), fever (21%), and decreased neutrophil count and fatigue (20%). Nineteen patients (31%) reported the most common grade ≥3 TRAEs. All grade ≥3 AEs occurred in the 2.5 mg / kg cohort. Among the 19 patients who underwent tumor evaluation in Phase Ia, the ORR was 5% (1 / 19), and the DCR was 32% (6 / 19). In 27 patients with stage Ib disease who were evaluable for response, the best overall response rate (BOR) was 29.6%, with 8 confirmed partial responses (PRs) and 12 stable disease (SDs). The ORR was 40% and DCR was 100% for patients with SCCHN, 44% and DCR was 89% for patients with NPC, and 0% and DCR was 25% for patients with CRC.

[0114] Conclusion: Compared with FDA-approved EGFR-targeted monoclonal antibodies (including cetuximab and panitumumab), the incidence of rash, pruritus, and other skin toxicities during MYK-3 treatment was lower than with cetuximab and panitumumab. MYK-3 demonstrated manageable safety and promising antitumor activity in phase I clinical trials, particularly in patients with EGFR-positive advanced solid tumors (including NPC and SCCHN) that had recurred or metastasized after multiple prior antitumor therapies. These results will be further validated in phase II trials. Based on patient safety data from the phase Ia dose-escalation and phase Ib dose-expansion phases, MYK-3 is expected to have improved safety and tolerability at a dose of 2.0 mg / kg compared to 2.5 mg / kg. Therefore, the phase II trial will further explore the possibility of establishing 2.0 mg / kg and 2.3 mg / kg as the recommended clinical doses for MYK-3.

[0115] 2) Clinical phase II Research method: An ongoing Phase II expansion study is investigating the treatment of recurrent metastatic nasopharyngeal carcinoma with MYK-3. Patients received MYK-3 once every three weeks (Q3W) at doses of 2.0 mg / kg and 2.3 mg / kg.

[0116] Exam Description: This clinical trial is evaluating the potential efficacy and safety of MYK-3 treatment in subjects with recurrent or metastatic nasopharyngeal carcinoma (NPC).

[0117] Research Subjects: The study consists of two parts: Part A (an exploratory Phase IIa study to confirm the efficacy of MYK-3 in NPC subjects eligible for second-line or greater therapy) and Part B (a confirmatory Phase IIb study in NPC subjects eligible for third-line or greater therapy).

[0118] All subjects were patients with recurrent metastatic nasopharyngeal carcinoma who were inoperable or ineligible for radiation therapy and had the following characteristics: (1) Part A: Patients who relapsed after initial surgery and radiation therapy, were ineligible for further surgery or radiation therapy, and had previously received at least first-line systemic treatment with a platinum-containing regimen and a PD-1 / PD-L1 inhibitor, with disease progression during treatment, recurrence after treatment, or intolerance. (2) Part B: Patients who had previously received at least second-line systemic chemotherapy and a PD-1 / PD-L1 inhibitor but failed. Previous chemotherapy regimens must have included a platinum-containing regimen, gemcitabine, or a taxane / capecitabine (platinum-containing regimens include cisplatin, carboplatin, and nedaplatin, and taxanes include paclitaxel, docetaxel, albumin-bound paclitaxel, and liposomal paclitaxel). (3) The disease was advanced, incurable metastatic tumor, and clinical trial participation was an appropriate treatment option. (4) Meet all inclusion criteria and do not meet any exclusion criteria. Platinum-containing regimens include cisplatin, carboplatin, and nedaplatin.

[0119] Research design: This Phase II trial consists of Parts A and B.

[0120] The exploratory phase of Part A will evaluate the safety and PK characteristics of MYK-3 in patients who are candidates for second-line or higher treatment, identify optimal dosing and tolerability, and verify the efficacy and antitumor activity of MYK-3 based on ORR assessed by an Independent Review Committee (IRC) using RECIST v1.1. If the safety and antitumor activity of MYK-3 are sufficiently demonstrated based on the recorded ORR in Part A, the study will proceed to Part B.

[0121] Part B is a randomized controlled clinical trial comparing the efficacy and safety of injectable MYK-3 with capecitabine / docetaxel in patients with recurrent metastatic nasopharyngeal carcinoma who have previously failed at least second-line systemic chemotherapy and PD-1 / PD-L1 therapy. This study will be stratified by two factors: liver metastasis (yes vs. no) and ECOG PS (0 vs. 1). Subjects in the experimental group will receive MYK-3 therapy, while subjects in the control group will receive capecitabine if they have not received capecitabine therapy in the past. If they have received capecitabine therapy in the past, they will receive either capecitabine or docetaxel at the investigator's discretion.

[0122] Based on the completed first-in-human data and subsequent analyses of efficacy, safety, PK, etc., the MTD was demonstrated to be 2.5 mg / kg in the Phase Ia dose-escalation phase. In the Phase Ib expansion phase, this dose level demonstrated favorable efficacy in the head and neck squamous cell carcinoma cohort and the nasopharyngeal carcinoma cohort. To evaluate the optimal dose for further development, taking into account the risk-benefit balance, this Phase II study selected doses below the MTD of 2.0 mg / kg and an intermediate dose of 2.3 mg / kg for further exploration in Part A of this study.

[0123] Part A: Subjects received 2.0 mg / kg or 2.3 mg / kg of MYK-3 intravenously on day 1 of every 3 weeks (Q3W, 3 weeks being one treatment cycle).

[0124] Part B: Subjects in the test group will receive 2.3 mg / kg of MYK-3 intravenously on day 1 of every 3 weeks (Q3W, 3 weeks = 1 treatment cycle). Subjects in the control group will receive 1000 mg / m on days 1-14 of every 3 weeks (Q3W, 3 weeks = 1 treatment cycle) at the investigator's option. 2 Oral capecitabine bid (if the first capecitabine dose is given on the night of D1, oral administration will continue until the morning of D15) or 75 mg / m bid on the first day of every 3 weeks (Q3W, 3 weeks is one treatment cycle). 2 of docetaxel was administered intravenously.

[0125] Based on comprehensive data on long-term administration safety, efficacy, PK, etc. from Part A and Phase I trials, the Safety Monitoring Committee (SMC) finally determined the recommended dose of MYK-3 in Part B to be 2.3 mg / kg.

[0126] Test results: The test results are shown in Tables 1, 2 and 3 below.

[0127] [Table 1]

[0128] [Table 2]

[0129] [Table 3]

[0130] Conclusion: An exploratory phase IIa study of MYK-3 demonstrated significant antitumor activity in patients with recurrent or metastatic nasopharyngeal carcinoma (NPC) who had previously received multiple tumor-related therapies. The objective tumor response rates at the 2.0 mg / kg and 2.3 mg / kg doses were 39.3% and 55.2%, respectively, offering new hope for this patient population, which currently lacks effective treatments. Because the 2.3 mg / kg dose group demonstrated superior therapeutic efficacy compared to the 2.0 mg / kg dose group and was also safe and well tolerated, 2.3 mg / kg was ultimately selected as the recommended dose for the treatment of recurrent and metastatic NPC with MYK-3.

Claims

1. 1. An antibody-drug conjugate having the structure shown in Formula I, or a pharmaceutically acceptable salt, solvate, or solvate of said salt, for treating a disease associated with epidermal growth factor receptor (EGFR). Ab-(LD) ｐ Formula I (In the formula, Ab represents an anti-epidermal growth factor receptor antibody, the anti-epidermal growth factor receptor antibody comprising a heavy chain and a light chain, wherein CDR1, CDR2, and CDR3 of the heavy chain variable region comprise the sequences shown in SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7, respectively, or variants thereof, and CDR1, CDR2, and CDR3 of the light chain variable region comprise the sequences shown in SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 14, respectively, or variants thereof; L represents a linker, and the linker is 6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (MC-vc-PAB); D represents a cytotoxic agent, said cytotoxic agent being MMAE; p is any number selected from 1 to 8, The antibody-drug conjugate, or a pharmaceutically acceptable salt, solvate, or solvate of said salt thereof, is administered at a total dose of 2.1 to 2.4 mg / kg every three weeks.

2. the antibody-drug conjugate, or a pharmaceutically acceptable salt, solvate, or solvate of said salt thereof, is administered at a total dose of 2.2 mg / kg to 2.4 mg / kg every three weeks; The antibody-drug conjugate, or a pharmaceutically acceptable salt, solvate, or solvate of the salt thereof, according to claim 1, characterized in that the antibody-drug conjugate, or a pharmaceutically acceptable salt, solvate, or solvate of the salt thereof is preferably administered at a total dose of 2.3 mg / kg every three weeks.

3. The antibody-drug conjugate, or the pharmacologically acceptable salt, solvate, or solvate of the salt thereof according to any one of claims 1 to 2, wherein a single dose of the antibody-drug conjugate, or the pharmacologically acceptable salt, solvate, or solvate of the salt thereof is 2.1 to 2.4 mg / kg (preferably 2.2 to 2.4 mg / kg, more preferably 2.3 mg / kg), and the administration frequency is once every three weeks.

4. The antibody-drug conjugate, or the pharmacologically acceptable salt, solvate, or solvate of the salt thereof according to any one of claims 1 to 2, wherein a single dose of the antibody-drug conjugate, or the pharmacologically acceptable salt, solvate, or solvate of the salt thereof is 0.7 to 1.4 mg / kg, and the administration frequency is once every 7 to 10 days.

5. The antibody-drug conjugate, or the pharmacologically acceptable salt, solvate, or solvate of the salt thereof according to any one of claims 1 to 4, wherein the antibody-drug conjugate, or the pharmacologically acceptable salt, solvate, or solvate of the salt thereof is administered by intravenous infusion.

6. the subject to which the antibody-drug conjugate, or the pharmacologically acceptable salt, solvate, or solvate of the salt thereof is to be administered is a human; Preferably, the subject to be administered with the antibody-drug conjugate, or a pharmacologically acceptable salt, solvate, or solvate of the salt thereof is a patient with a disease associated with epidermal growth factor receptor (EGFR), Preferably, the subject to be administered the antibody-drug conjugate, or a pharmacologically acceptable salt, solvate, or solvate of the salt thereof is a patient who has previously received treatment with an anti-EGFR drug, and preferably, the anti-EGFR drug is an anti-EGFR antibody; The antibody-drug conjugate, or the pharmacologically acceptable salt, solvate, or solvate of the salt thereof according to any one of claims 1 to 5, wherein the subject to be administered with the antibody-drug conjugate, or the pharmacologically acceptable salt, solvate, or solvate of the salt thereof is a patient who overexpresses EGFR.

7. the disease associated with epidermal growth factor receptor (EGFR) is cancer; Preferably, the cancer is selected from nasopharyngeal cancer, head and neck squamous cell carcinoma, biliary tract cancer, esophageal cancer, duodenal cancer, colorectal cancer, colon cancer, rectal cancer, head and neck cancer, lung cancer (e.g., non-small cell lung cancer), ovarian cancer, cervical cancer, bladder cancer, esophageal cancer, breast cancer, kidney cancer, prostate cancer, gastric cancer, pancreatic cancer, and glioma; Preferably, the cancer is selected from nasopharyngeal cancer, head and neck squamous cell carcinoma, non-small cell lung cancer, biliary tract cancer, gastric cancer, esophageal cancer, and duodenal cancer; The antibody-drug conjugate, or a pharmacologically acceptable salt, solvate, or solvate of the salt thereof, according to claim 1 or 6, characterized in that the cancer is preferably selected from EGFR-positive, HER2-negative advanced gastric cancer, recurrent or metastatic nasopharyngeal carcinoma, recurrent or metastatic head and neck squamous cell carcinoma, EGFR-positive advanced biliary adenocarcinoma, and EGFR-positive advanced non-small cell lung cancer.

8. p is any number selected from 2 to 6, The antibody-drug conjugate, or a pharmacologically acceptable salt, solvate, or solvate of said salt thereof, according to any one of claims 1 to 7, characterized in that p is preferably any number selected from 3 to 5.

9. The anti-epidermal growth factor receptor antibody has one or more of the following characteristics: 1) the FR1, FR2, FR3, and FR4 regions of the heavy chain variable region of the anti-epidermal growth factor receptor antibody comprise the sequences shown in SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and SEQ ID NO: 11, respectively, or variants thereof; 2) the FR1, FR2, FR3, and FR4 regions of the light chain variable region of the anti-epidermal growth factor receptor antibody comprise the sequences shown in SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, and SEQ ID NO: 18, respectively, or variants thereof; 3) the heavy chain constant region of the anti-epidermal growth factor receptor antibody is selected from human IgG, IgM, IgA, IgD, and IgE constant regions or mutants of these constant regions; Preferably, the IgG is selected from IgG1, IgG2, IgG3, and IgG4; 4) The antibody-drug conjugate according to any one of claims 1 to 8, wherein the light chain constant region of the anti-epidermal growth factor receptor antibody is a human-derived lambda constant region, a human kappa constant region, or a mutant of these constant regions, or a pharmacologically acceptable salt, solvate, or solvate of the salt thereof.

10. The anti-epidermal growth factor receptor antibody has one or more of the following characteristics: 1) the sequence of the heavy chain variable region of the anti-epidermal growth factor receptor antibody comprises the sequence shown in SEQ ID NO: 1 or a sequence having greater than 70%, preferably greater than 75%, 80%, 85%, 90%, 95%, or 99% homology to the sequence shown in SEQ ID NO: 1; Preferably, the sequence of the heavy chain variable region of the anti-epidermal growth factor receptor antibody is set forth in SEQ ID NO: 1, 2) the sequence of the light chain variable region of the anti-epidermal growth factor receptor antibody comprises the sequence shown in SEQ ID NO: 2 or a sequence having greater than 70%, preferably greater than 75%, 80%, 85%, 90%, 95%, or 99% homology to the sequence shown in SEQ ID NO: 2; Preferably, the sequence of the light chain variable region of the anti-epidermal growth factor receptor antibody is set forth in SEQ ID NO: 2, 3) the sequence of the heavy chain constant region of the anti-epidermal growth factor receptor antibody comprises the sequence shown in SEQ ID NO: 3 or a sequence having greater than 70%, preferably greater than 75%, 80%, 85%, 90%, 95%, or 99% homology to the sequence shown in SEQ ID NO: 3; Preferably, the sequence of the heavy chain constant region of the anti-epidermal growth factor receptor antibody is set forth in SEQ ID NO: 3, 4) the sequence of the light chain constant region of the anti-epidermal growth factor receptor antibody comprises the sequence shown in SEQ ID NO: 4 or a sequence having more than 70%, preferably more than 75%, 80%, 85%, 90%, 95%, or 99% homology to the sequence shown in SEQ ID NO: 4; The antibody-drug conjugate according to any one of claims 1 to 9, wherein the sequence of the light chain constant region of the anti-epidermal growth factor receptor antibody is preferably that shown in SEQ ID NO: 4, or a pharmacologically acceptable salt, solvate, or solvate of said salt thereof.

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