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KR-20260066104-A - Antibody-drug conjugate targeting GPC3 and its applications

KR20260066104AKR 20260066104 AKR20260066104 AKR 20260066104AKR-20260066104-A

Abstract

The present invention relates to a GPC3 antibody-drug conjugate and its applications, specifically providing an antibody-drug conjugate, or a pharmaceutically acceptable salt, solvate, or solvate of the salt thereof. The antibody-drug conjugate has a structure represented by Formula I, wherein Ab is an anti-GPC3 antibody. The antibody-drug conjugate of the present invention exhibits good tumor cell growth inhibitory activity in vivo and in vitro and has promising application prospects. Ab-(LD) p Formula I

Inventors

  • 리, 홍펑
  • 팡, 레이
  • 왕, 옌춘
  • 리우, 원차오
  • 리우, 웨이
  • 마, 나
  • 후, 차오홍
  • 선, 하오
  • 추이, 페이페이
  • 양, 리우

Assignees

  • 레푸 바이오파마 컴퍼니 리미티드

Dates

Publication Date
20260512
Application Date
20240904
Priority Date
20230905

Claims (12)

  1. As an antibody-drug conjugate, or a pharmaceutically acceptable salt, solvate, or solvate of said salt thereof, The above antibody-drug conjugate has a structure represented by Formula I, and Ab-(LD) p Formula I In the above equation I, Ab is an anti-GPC3 antibody or an antigen-binding fragment thereof, wherein the anti-GPC3 antibody comprises a heavy chain variable region and a light chain variable region, and the heavy chain variable regions CDR1, CDR2, and CDR3 each comprise a sequence represented by SEQ ID NO. 1, SEQ ID NO. 2, and SEQ ID NO. 3 or a mutant thereof, and the light chain variable regions CDR1, CDR2, and CDR3 each comprise a sequence represented by SEQ ID NO. 9, SEQ ID NO. 10, and SEQ ID NO. 11 or a mutant thereof; L is a linker selected from mc-AAN, mc-AAQ, mcc-AAN, mcc-AAQ, mcc-Ala-Ala-(3-cyano-alanine), mcc-Ala-Ala-(2-amino-4-cyanobutanoic acid), mcc-Ala-Ala-Ala-Ala-Gln; D is a cytotoxin called exatecan; p is an arbitrary value between 1 and 10; An antibody-drug conjugate in which the mcc or mc end of the L linker is covalently bonded to Ab and the other end of the L linker is covalently bonded to D, or a pharmaceutically acceptable salt, solvate, or solvate of said salt thereof.
  2. In paragraph 1, The structural formula of LD before conjugation with Ab is as follows: Each LD is an antibody-drug conjugate conjugated to the mercapto of Ab through terminal succinimide, or a pharmaceutically acceptable salt, solvate, or solvate of said salt thereof.
  3. In any one of paragraphs 1 to 2, The antibody-drug conjugate is an antibody-drug conjugate having a structure represented by formulas (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), or (IIi), or a pharmaceutically acceptable salt, solvate, or solvate of said salt thereof: (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi).
  4. In any one of paragraphs 1 through 3, The above anti-GPC3 antibody has one or more selected from technical features (1) to technical features (2), and In technical feature (1), the heavy chain variable regions FR1, FR2, FR3, and FR4 of the anti-GPC3 antibody each comprise sequences represented by SEQ ID NO. 4, SEQ ID NO. 5, SEQ ID NO. 6, and SEQ ID NO. 7 or mutants thereof; In technical feature (2), the light chain variable regions FR1, FR2, FR3, and FR4 of the anti-GPC3 antibody each comprise sequences represented by SEQ ID NO. 12, SEQ ID NO. 13, SEQ ID NO. 14, and SEQ ID NO. 15 or mutants thereof, or the light chain variable regions FR1, FR2, FR3, and FR4 of the anti-GPC3 antibody each comprise sequences represented by SEQ ID NO. 12, SEQ ID NO. 13, SEQ ID NO. 17, and SEQ ID NO. 15 or mutants thereof, antibody-drug conjugate, or pharmaceutically acceptable salts, solvates, or solvates of said salts thereof.
  5. In any one of paragraphs 1 through 4, The above anti-GPC3 antibody has one or more selected from technical features (1) to technical features (2), and In technical feature (1), the heavy chain variable region of the anti-GPC3 antibody comprises the sequence represented by SEQ ID NO. 8 or a mutant thereof; In technical feature (2), the light chain variable region of the anti-GPC3 antibody comprises an antibody-drug conjugate comprising the sequence represented by SEQ ID NO. 16 or SEQ ID NO. 18 or a mutant thereof, or a pharmaceutically acceptable salt, solvate thereof, or solvate of said salt.
  6. In any one of paragraphs 1 through 5, The above anti-GPC3 antibody has one or more selected from technical features (1) to technical features (2), and In technical feature (1), the heavy chain constant region of the anti-GPC3 antibody is selected from human IgG, IgM, IgA, IgD, IgE constant regions or mutants of said constant region; Preferably, the IgG is selected from IgG1, IgG2, IgG3, and IgG4; In technical feature (2), the light chain constant region of the anti-GPC3 antibody is an antibody-drug conjugate selected from a human lambda constant region, a kappa constant region or a mutant of said constant region, or a pharmaceutically acceptable salt, solvate, or solvate of said salt.
  7. In any one of paragraphs 1 through 6, The above anti-GPC3 antibody has one or more selected from technical features (1) to technical features (2), and In technical feature (1), the heavy chain constant region of the anti-GPC3 antibody comprises a sequence represented by SEQ ID NO. 19, or comprises a sequence having homology with the sequence represented by SEQ ID NO. 19 greater than 70%, preferably greater than 75%, 80%, 85%, 90%, 95%, or 99%; Preferably, the sequence of the heavy chain constant region of the anti-GPC3 antibody is denoted by SEQ ID NO. 19; In technical feature (2), the light chain constant region of the anti-GPC3 antibody comprises a sequence represented by SEQ ID NO. 20, or comprises a sequence having homology with the sequence represented by SEQ ID NO. 20 greater than 70%, preferably greater than 75%, 80%, 85%, 90%, 95%, or 99%; Preferably, the sequence of the light chain constant region of the anti-GPC3 antibody is the antibody-drug conjugate represented by SEQ ID NO. 20, or a pharmaceutically acceptable salt, solvate, or solvate of the same.
  8. In any one of paragraphs 1 through 7, p is an arbitrary value between 2 and 8; Preferably, p is an antibody-drug conjugate having any value between 4 and 8 (e.g., 4.0, 7.9, or 8.0), or a pharmaceutically acceptable salt, solvate, or solvate of the same.
  9. As a pharmaceutical composition, An antibody-drug conjugate according to any one of claims 1 to 8, or comprising a pharmaceutically acceptable salt, solvate, or solvate of said salt thereof; Optionally, the pharmaceutical composition further comprises at least one pharmaceutical auxiliary substance; Or optionally, the pharmaceutical composition further comprises at least one of a chemotherapy agent, an immunotherapy agent, and an immunosuppressant for tumor treatment.
  10. In the manufacture of drugs, for use of an antibody-drug conjugate according to any one of claims 1 to 8, or a pharmaceutically acceptable salt, solvate, or solvate of said salt, or a pharmaceutical composition according to claim 9, The above drug is used for the prevention and/or treatment of GPC3-positive related diseases; Preferably, the GPC3-positive related disease is selected from liver cancer, lung cancer, gastric cancer, head and neck cancer, esophageal cancer, Merkel cell carcinoma, liposarcoma, breast cancer, ovarian cancer, melanoma, squamous cell carcinoma, renal cell carcinoma, pancreatic cancer, prostate cancer, colorectal cancer, glioma, kidney cancer, bladder cancer, cervical cancer, gallbladder cancer, and glioblastoma; More preferably, the above-mentioned GPC3-positive related disease is liver cancer.
  11. As a method for non-therapeutic inhibition of tumor angiogenesis in vitro, delaying tumor progression, inhibiting tumor growth, or inhibiting tumor cell proliferation, The method comprises the step of contacting a tumor cell with an antibody-drug conjugate according to any one of claims 1 to 8, or a pharmaceutically acceptable salt, solvate, or solvate of said salt, or a pharmaceutical composition according to claim 9, wherein the tumor is a GPC3-expressing tumor; Preferably, the GPC3-expressing tumor is selected from liver cancer, lung cancer, gastric cancer, head and neck cancer, esophageal cancer, Merkel cell carcinoma, liposarcoma, breast cancer, ovarian cancer, melanoma, squamous cell carcinoma, renal cell carcinoma, pancreatic cancer, prostate cancer, colorectal cancer, glioma, kidney cancer, bladder cancer, cervical cancer, gallbladder cancer, and glioblastoma; More preferably, the method in which the GPC3-expressing tumor is liver cancer.
  12. As a method for the treatment and/or prevention of GPC3-positive related diseases, The method comprises the step of administering to a subject in need a therapeutically and/or prophylactically effective amount of an antibody-drug conjugate according to any one of claims 1 to 8, or a pharmaceutically acceptable salt, solvate, or solvate of said salt, or a pharmaceutical composition according to claim 9; Preferably, the GPC3-positive related disease is selected from liver cancer, lung cancer, gastric cancer, head and neck cancer, esophageal cancer, Merkel cell carcinoma, liposarcoma, breast cancer, ovarian cancer, melanoma, squamous cell carcinoma, renal cell carcinoma, pancreatic cancer, prostate cancer, colorectal cancer, glioma, kidney cancer, bladder cancer, cervical cancer, gallbladder cancer, and glioblastoma; More preferably, the above-mentioned GPC3-positive related disease is liver cancer.

Description

Antibody-drug conjugate targeting GPC3 and its applications Cross-reference of related applications The present application claims all priority based on the application with CN application number 202311141587.3 and the filing date September 5, 2023, and the application with CN application number 202411179009.3 and the filing date August 26, 2024, and the contents of all CN applications are incorporated by reference into the present application. The present invention relates to the field of pharmaceutical chemistry, specifically to a GPC3 antibody-drug conjugate and its applications. Liver cancer is the fifth most common cancer worldwide and the third most common cause of cancer-related deaths. Hepatocellular carcinoma (HCC) is the major form of liver cancer, accounting for 90% of all liver cancers. Currently, the approved treatment is a multidisciplinary comprehensive approach centered on surgical resection. While short-term treatment efficacy has improved, long-term efficacy remains unsatisfactory. Therefore, seeking new treatment strategies that are more effective for liver cancer is currently a research hotspot both domestically and internationally, and the development of new drugs for liver cancer treatment remains an urgent priority. The formal name of GPC3 is Glypican-3. It is a member of the heparan sulfate proteoglycan family and is anchored to the cell surface by glycosylphosphatidylinositol on the cell membrane. The protein core of GPC3 consists of two subunits, with the N-terminal subunit measuring approximately 40 kDa and the C-terminal subunit measuring approximately 30 kDa. Six glypicans (GPC1-6) have been identified in mammals. GPC3 plays an important role in regulating cell proliferation, differentiation, adhesion, and migration. Some studies suggest that GPC3 interacts with Wnt proteins and Frizzled receptors to form complexes and trigger downstream signaling. The core protein of GPC3 can serve as a Wnt co-receptor or receiver. The expression of the GPC3 protein derived from the GPC3 gene in the human body varies significantly across different developmental stages and tissues. For example, it is present in very low amounts in normal adult tissues and is low or absent in cancers such as gastric, breast, and ovarian cancers, whereas it is often overexpressed in hepatocellular carcinoma (HCC) (positive rate over 70%). Studies have shown that GPC3 is a liver cancer-specific associated antigen that can help identify lesion characteristics in the early stages of HCC and significantly improve diagnostic accuracy. Therefore, GPC3 is considered to have great potential for targeted therapy in liver cancer. Recently, interest in GPC3-targeted therapies has been increasing. Currently, therapeutic strategies targeting GPC3 are primarily focused on antibody drugs, cell therapies, and vaccines. Therapeutic anti-GPC3 antibodies have been developed, including antibodies such as GC33 (completed Phase 2 clinical trials), YP7, and HN3, as well as bispecific antibodies currently in clinical trials such as ERY974 and CM350. Some of these antibodies may act by inhibiting Wnt signaling in liver cancer cells. In addition, various studies have developed chimeric antigen receptor (CAR) T-cell immunotherapy targeting GPC3 for the treatment of liver cancer. Studies using mouse models with xenografts or orthotopic liver cancer have revealed that CAR-T cells can eliminate GPC3-positive cancer cells by inducing apoptosis induced by perforin and granzyme and inhibiting WNT signaling in tumor cells. Figure 1 shows the affinity of each GPC3 antibody for GPC3-CHO-K1 cells. Figure 2 shows the affinity of each GPC3 antibody for HepG2 cells. Figure 3 shows the affinity of each GPC3 antibody for Huh-7 cells. Figure 4 shows the internalization of each GPC3 antibody in HepG2 cells. Figure 5 shows the hydrophobic interaction chromatogram of Hu 52H5D3B8-8-mc-AAN-Exatecan(ADC-1). Figure 6 shows the hydrophobic interaction chromatogram of Hu 52H5D3B8-8-mcc-AAN-Exatecan(ADC-2). Figure 7 shows the hydrophobic interaction chromatogram of Hu 52H5D3B8-8-mc-AAQ-Exatecan (ADC-3). Figure 8 shows the hydrophobic interaction chromatogram of Hu 52H5D3B8-8-mcc-AAQ-Exatecan (ADC-4). Figure 9 shows the hydrophobic interaction chromatogram of Hu 52H5D3B8-7-mcc-AAQ-Exatecan (ADC-5). Figure 10 shows the hydrophobic interaction chromatogram of Hu 52H5D3B8-8-mcc-AAQ-Exatecan (ADC-6). Figure 11 shows the hydrophobic interaction chromatogram of Hu 52H5D3B8-7-mcc-AAQ-Exatecan (ADC-7). Figure 12 shows the hydrophobic interaction chromatogram of Hu 52H5D3B8-8-mcc-AA-(3-cyano-L-alanine)-Exatecan(ADC-8). Figure 13 shows the hydrophobic interaction chromatogram of Hu 52H5D3B8-8-mcc-AAAAQ-Exatecan (ADC-9). Figure 14 shows the affinity of GPC3 antibody and ADC for Huh7 cells. Figure 15 shows the results of endocytosis of GPC3 antibody and ADC in Huh7 cells. Figures 16–18 show representative diagrams illustrating the killing of different GPC3-ADCs on Huh7 cells. Fi