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KR-20260068097-A - Bispecific antibody binding to CD3 and ganglioside NGcGM3

KR20260068097AKR 20260068097 AKR20260068097 AKR 20260068097AKR-20260068097-A

Abstract

The present invention relates to the fields of biotechnology and immuno-oncology. Disclosed are bispecific antibodies comprising an antibody, antibody fragment, or short-chain variable fragment that recognizes NGcGM3 gangliosides on tumor cells, and an antibody, antibody fragment, or short-chain variable fragment that recognizes CD3 antigens on human immune effector cells. These bispecific antibodies are characterized by their efficacy in mediating selective cytotoxicity against NGcGM3-positive tumor cells but not against normal cells capable of expressing these gangliosides, and in recruiting T lymphocytes as well as NKTs. The bispecific antibodies of the present invention and the nucleic acids encoding them are useful for the treatment of lymphoproliferative diseases and solid tumors expressing NGcGM3.

Inventors

  • 에르난데스 가르시아, 테이스
  • 플라센시아 이글레시아스, 클라우디아 아라셀리스
  • 디에고 마토스, 아나 카를라
  • 레온 몬손, 칼렛
  • 제크리, 라티파
  • 융, 군드람

Assignees

  • 센트로 데 인뮤놀러지아 모레큘러

Dates

Publication Date
20260513
Application Date
20240823
Priority Date
20230908

Claims (13)

  1. A bispecific antibody comprising an antibody, antibody fragment, or single chain variable fragment (scFv) that recognizes NGcGM3 gangliosides on tumor cells; and an antibody, antibody fragment, or scFv that recognizes human CD3 molecules.
  2. In paragraph 1, the antibody recognizing NGcGM3 is a bispecific antibody that is human IgG.
  3. In claim 1, the antibody is a bispecific antibody having a format selected from the group including: TanDab, DART, DART-Fc, DuoBody, CrossMab, KiH, BiTE, Triomab, IgG-scFv.
  4. In paragraph 2, a bispecific antibody in which the heavy chain is fused directly or through a peptide linker to an antibody, antibody fragment, or scFv specific to human CD3.
  5. In claim 4, the antibody, antibody fragment, or scFv that recognizes a human CD3 molecule is a bispecific antibody having a sequence selected from the group comprising the sequence of SEQ ID NOs 5 to 7, or a variant thereof having more than 90% identity with said sequence.
  6. A bispecific antibody according to claim 4, wherein the antibody, antibody fragment, or scFv recognizing NGcGM3 has a heavy chain variable region selected from a group comprising the sequences of SEQ ID NO. 10 and SEQ ID NO. 15, or variants thereof having more than 90% identity with these sequences.
  7. A bispecific antibody according to claim 4, wherein the antibody, antibody fragment, or scFv recognizing NGcGM3 has a light chain variable region selected from a group comprising the sequences of SEQ ID NOs 11 to 14, or variants thereof having more than 90% identity with said sequences.
  8. A bispecific antibody according to paragraph 2, wherein the human IgG sequence is selected from a group comprising the sequences of SEQ ID NOs 1 to 4 or variants thereof having more than 90% identity with the sequences.
  9. A pharmaceutical composition comprising, as an active ingredient, a bispecific antibody of any one of claims 1 to 8, and a pharmaceutically acceptable carrier.
  10. Use of a bispecific antibody of any one of claims 1 to 8 for treating lymphoproliferative diseases and solid tumors expressing NGcGM3.
  11. Use of a nucleic acid encoding BAb-T according to any one of claims 1 to 8 in gene therapy for tumors expressing NGcGM3.
  12. In claim 11, the gene therapy is based on the injection of mRNA encoding BAb-T or gene carrier particles.
  13. A treatment method comprising administering a bispecific antibody of any one of claims 1 to 8 to a subject requiring treatment via a subcutaneous, intravenous, intradermal, intramuscular, intratumoral, or intraperitoneal route in a dose range of 20 µg to 10 mg.

Description

Bispecific antibody binding to CD3 and ganglioside NGcGM3 The present invention relates to the fields of biotechnology and immuno-oncology. Specifically, it discloses a bispecific antibody that recognizes an NGcGM3 antigen expressed in a tumor and a CD3 molecule having the ability to recruit effector functions of T, NKT, and Tγδ cells. T cell-engaging bispecific antibodies (BAb-T) are increasingly establishing themselves as one of the most promising alternatives for the redirection of effector cells of the immune system toward tumors, where they recognize antigens on the surface of malignant cells (Wu, Z. and N. Cheung (2018) Pharmacology therapeutics 182:161-175). As such, the recruitment of T lymphocytes toward tumor cells, independent of the specific recognition of antigens by T cell receptors, is a very attractive and novel approach. One of the challenges of this type of therapy is the appropriate selection of tumor antigens to target. Tumor-specific antigens derived from genetic and epigenetic changes are those that selectively and maximally target tumors (Apavaloaei, A., et al. (2020) Cancers 12 (9):2607). Unfortunately, many of them are intracellular and are not accessible to standard BAb-T. Tumor-selective antigens include those overexpressed by tumors or located differently from normal tissues, which are also used in BAb-T-based therapies, but do not rule out the emergence of potential side effects. In particular, BAb-T for solid tumors often presents safety issues associated with cytokine release syndrome (CRS) and damages normal tissues expressing antigens. Preliminary results for BAb-T in solid tumors have reported overall higher CRS rates (19–91%) compared to those for the treatment of hematological malignancies such as blinatumumab (7–15%) (Bendell, J. C., et al. (2020); J Clin Oncol 38: 55-52; Borghaei, H., et al. (2020); Middleton, M. R., et al. (2020); Clinical Cancer Research 26 (22): 5869-5878; Tran, B., et al (2020). Annals of Oncology 31: S507). Toxicity to extratumor targets was a limiting factor in the use of BAb-T for this type of tumor (Kobold, S., et al. (2018) Frontiers in Oncology 8:285). NGcGM3 gangliosides have become a widely studied target due to their expression in human tumors of various sites and their near absence in normal tissues (Bardor, M., et al. (2005) Journal of Biological Chemistry 280(6): 4228-4237), and several therapies based on them have been conceived (Blanco, R., et al. (2011) International Scholarly Research Notices; Lahera, T., et al. (2014) Oncol). This includes vaccines such as Glycovax (Carr, A., et al. (2003) Journal of Clinical Oncology 21(6): 1015-1021), monoclonal antibodies (MAbs) such as 14F7, which feature fine recognition capable of distinguishing between the N-acetylated version (not recognized) of GM3 gangliosides and the N-glycosylated variant (recognized) (Carr, A., et al. (2002). Hybridoma hybridomics 21(6): 463-468; Casadesus, A. V., et al. (2013) Glycoconjugate 30: 687-699), and fragments derived from them such as 3FM and 8Bhl (Rojas, G., et al. (2004). Journal of immunological methods 293(1-2): This includes the development of 71-83). The 14F7 antibody can induce oncosis-like cell death in mouse L1210 tumor cells (lymphoid-derived), but not in normal mouse lymphocytes (B and TCD4+). This difference in cytotoxicity may be attributed to the distinct distribution of NGcGM3 gangliosides in the lipid rafts of the plasma membrane between normal and tumor cells (Roque-Navarro, L., et al. (2008) Molecular Cancer Therapeutics 7(7): 2033-2041). However, this difference between tumor and normal cells in terms of ganglioside distribution does not necessarily mean that similar selectivity for other types of death induced by the 14F7 binding domain and leading to the participation of effector cells is inevitable. Given the non-protein nature of NGcGM3, therapeutic approaches such as Mab and vaccines are inefficient in recruiting T cell responses and cytotoxic capabilities against tumors expressing this ganglioside. This increases the need to develop therapies that mediate T-lymphocyte cytotoxicity against NGcGM3-positive tumor cells. To date, no BAb-T specific to this ganglioside is known. Since the first generation BAb-T, various formats ranging from the smallest designs to those based on a complete IgG structure have been known (Godar, M. et al. (2018). Expert Opinion on Therapeutic patents 28(3):251-276). Although the selected format may affect the functionality of these antibodies, the key role of the target molecule in the efficacy and safety of BAb-T is well known (Li, H., et al. (2020) Cellular molecular immunology 17(5): 451-461). This is influenced by the location of the recognized epitope and the size of the target molecule. These characteristics play a key role in the efficiency of immune synapse formation and subsequent T lymphocyte activation, and are determinants of the cytotoxic efficacy of this type of antibody (Li, J., et al. (2017).