GPRC5D antibody and its applications

Abstract

This invention provides antibodies and related biological agents that can be used in multiple myeloma patients who are B-cell maturation antigen (BCMA) negative or have low BCMA expression, and that can provide a better clinical response. [Solution] The present invention provides an antibody relating to G protein-coupled receptor class C group 5 member D (GPRC5D) having a specific sequence. Specifically, the present invention discloses an antibody or antigen-binding fragment thereof that specifically binds to GPRC5D, a nucleic acid encoding it, an expression vector and expression cells, a method of manufacture, a pharmaceutical composition, and their uses in the manufacture of a pharmaceutical composition for treating a disease, for example, in the treatment of tumors.

Claims

1. An antibody or antigen-binding fragment thereof that specifically binds to G protein-coupled receptor class C group 5 member D (GPRC5D), wherein the antibody or antigen-binding fragment comprises a heavy chain variable region and a light chain variable region, where, (1) The light chain variable region includes LCDR1, LCDR2, and LCDR3, wherein LCDR1 has any one of the LCDR1 sequences shown below or a sequence having 1, 2, 3 or more amino acid insertions, deletions, and / or substitutions compared to the aforementioned sequence, LCDR2 has any one of the LCDR2 sequences shown below or a sequence having 1, 2, 3 or more amino acid insertions, deletions, and / or substitutions compared to the aforementioned sequence, and LCDR3 has any one of the LCDR3 sequences shown below or a sequence having 1, 2, 3 or more amino acid insertions, deletions, and / or substitutions compared to the aforementioned sequence, Table 1 and, (2) The heavy chain variable region includes HCDR1, HCDR2 and HCDR3, wherein HCDR1 has any one of the following HCDR1 sequences or a sequence having 1, 2, 3 or more amino acid insertions, deletions and / or substitutions compared to the aforementioned sequence, HCDR2 has any one of the following HCDR2 sequences or a sequence having 1, 2, 3 or more amino acid insertions, deletions and / or substitutions compared to the aforementioned sequence, and HCDR3 has any one of the following HCDR3 sequences or a sequence having 1, 2, 3 or more amino acid insertions, deletions and / or substitutions compared to the aforementioned sequence, Table 2 Preferably, the antibody or its antigen-binding fragment is L1+H1, L2+H2, L3+H3, L4+H4, L5+H5, L6+H6, L7+H7, L8+H8, L9+H9, L10+H10, L11+H11, L12+H12, L13+H13, L14+H14, L15+H15, L16+H16, L3+H17, L3+H18, L5+H19, L5+H An antibody or antigen-binding fragment thereof comprising a sequence of six CDRs in a combination of light chain variable region and heavy chain variable region, namely L17+H13, L18+H13, L19+H13, L20+H13, or L21+H13, or a sequence of six CDRs having one, two, three, or more amino acid insertions, deletions, and / or substitutions compared to the sequence of the six CDRs.

2. (1) The light chain variable region sequence includes any one of the sequences shown in SEQ ID NO: 14, 16, 18, 20, 22, 24, 26, 28, 118-120, 130-131, 144-145, 160-163, 172-175, 184-187, 196-197, 206-208, 218-221 and 231-233, or a sequence having 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher identity with the sequence. and, (2) The heavy chain variable region sequence includes any one of the sequences shown in SEQ ID NO: 13, 15, 17, 19, 21, 23, 25, 27, 121-125, 132-137, 146-152, 164-166, 176-178, 188-190, 198-201, 209-212, 222 and 234-238, or a sequence having 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher identity with the sequence. Preferably, the antibody or its antigen-binding fragment has the following light chain variable region and heavy chain variable region, (1) The light chain variable region and the heavy chain variable region each include the sequences shown in SEQ ID NO: 14 and SEQ ID NO: 13, (2) The light chain variable region and the heavy chain variable region each include the sequences shown in SEQ ID NO: 16 and SEQ ID NO: 15, (3) The light chain variable region and the heavy chain variable region each include the sequences shown in SEQ ID NO: 18 and SEQ ID NO: 17, (4) The light chain variable region and the heavy chain variable region each include the sequences shown in SEQ ID NO: 20 and SEQ ID NO: 19, (5) The light chain variable region and the heavy chain variable region each include the sequences shown in SEQ ID NO: 22 and SEQ ID NO: 21, (6) The light chain variable region and the heavy chain variable region each include the sequences shown in SEQ ID NO: 24 and SEQ ID NO: 23, (7) The light chain variable region and the heavy chain variable region each include the sequences shown in SEQ ID NO: 26 and SEQ ID NO: 25, (8) The light chain variable region and the heavy chain variable region each include the sequences shown in SEQ ID NO: 28 and SEQ ID NO: 27, (9) The light chain variable region includes the sequence shown in any one of SEQ ID NO: 118 to 120, and the heavy chain variable region includes the sequence shown in any one of SEQ ID NO: 121 to 125. (10) The light chain variable region includes the sequence shown in any one of SEQ ID NO: 130 to 131, and the heavy chain variable region includes the sequence shown in any one of SEQ ID NO: 132 to 137. (11) The light chain variable region includes the sequence shown in any one of SEQ ID NO: 144 to 145, and the heavy chain variable region includes the sequence shown in any one of SEQ ID NO: 146 to 152. (12) The light chain variable region includes the sequence shown in any one of SEQ ID NO: 160 to 163, and the heavy chain variable region includes the sequence shown in any one of SEQ ID NO: 164 to 166. (13) The light chain variable region includes the sequence shown in any one of SEQ ID NO: 172 to 175, and the heavy chain variable region includes the sequence shown in any one of SEQ ID NO: 176 to 178. (14) The light chain variable region includes the sequence shown in any one of SEQ ID NO: 184 to 187, and the heavy chain variable region includes the sequence shown in any one of SEQ ID NO: 188 to 190. (15) The light chain variable region includes the sequence shown in any one of SEQ ID NO: 196 to 197, and the heavy chain variable region includes the sequence shown in any one of SEQ ID NO: 198 to 201. (16) The light chain variable region includes the sequence shown in any one of SEQ ID NO: 206 to 208, and the heavy chain variable region includes the sequence shown in any one of SEQ ID NO: 209 to 212. (17) The light chain variable region includes the sequence shown in any one of SEQ ID NO: 218 to 221, and the heavy chain variable region includes the sequence shown in SEQ ID NO:

222. (18) The light chain variable region includes the sequence shown in any one of SEQ ID NO: 231 to 233, and the heavy chain variable region includes the sequence shown in any one of SEQ ID NO: 234 to 238. Or (19) the antibody or antigen-binding fragment according to claim 1, wherein the light chain variable region includes a sequence having 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher identity with the light chain variable region shown in any one of (1) to (18), and the heavy chain variable region includes a sequence having 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher identity with the heavy chain variable region shown in any one of (1) to (18).

3. The antibody or antigen-binding fragment thereof according to claim 1 or 2, which is a chimeric, humanized, or entirely human-derived antibody.

4. An antibody or antigen-binding fragment thereof according to any one of claims 1 to 3, which can bind to human or monkey GPRC5D.

5. The antigen-binding fragment is F(ab) ２ An antibody or antigen-binding fragment thereof according to any one of claims 1 to 4, selected from one or more of Fab', Fab, Fv, scFv, nanoantibody, or affibody.

6. The antibody or its antigen-binding fragment further comprises a constant region sequence of any one of the human or mouse antibodies IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE, and IgD, preferably comprising a constant region sequence of a human or mouse antibody IgG1, IgG2, IgG3, or IgG4, or comprising a sequence having 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher identity with the constant region sequence of a human or mouse antibody IgG1, IgG2, IgG3, or IgG4, and further comprising the antibody or its antigen-binding fragment further comprises An antibody or antigen-binding fragment thereof according to any one of claims 1 to 5, wherein a therapeutic agent or tracer is conjugated, preferably the therapeutic agent is selected from radioisotopes, chemotherapeutic agents or immunomodulators, the tracer is selected from radiocontrast agents, paramagnetic ions, metals, fluorescent labels, chemiluminescent labels, ultrasonic contrast agents and photosensitizers, and more preferably the cytotoxic agent is selected from alkaloids, methotrexate, anthracycline antibiotics, taxanes, pyrrolobenzodiazepines or toxin compounds.

7. A multispecific antigen-binding molecule comprising an antibody or antigen-binding fragment thereof as described in any one of claims 1 to 6, and another antigen-binding molecule that binds to an antigen other than GPRC5D, or to a GPRC5D epitope different from the antibody or antigen-binding fragment described in any one of claims 1 to 6, Preferably, the other antigen-binding molecule is an antibody or an antigen-binding fragment thereof. Preferably, the multispecific antigen-binding molecule may be bispecific, triplicate, or quadruplicate. Preferably, the multispecific antigen-binding molecule may be divalent, trivalent, tetravalent, pentavalent, or hexavalent.

8. A chimeric antigen receptor (CAR), wherein the chimeric antigen receptor comprises at least a signal peptide, an extracellular antigen-binding domain, a hinge region, a transmembrane domain, and an intracellular signaling domain, and the extracellular antigen-binding domain comprises a GPRC5D antibody or its antigen-binding fragment according to any one of claims 1 to 6, or a multispecific antigen-binding molecule according to claim 7.

9. An immune effector cell, wherein the immune effector cell expresses the chimeric antigen receptor described in claim 8, or comprises a nucleic acid fragment encoding the chimeric antigen receptor described in claim 8, preferably the immune effector cell is selected from T cells, NK cells, NKT cells, DNT cells, monocytes, macrophages, dendritic cells, or mast cells, and the T cell is preferably selected from cytotoxic T cells (CTLs), regulatory T cells, or helper T cells, and preferably the immune effector cell is an autologous immune effector cell or an allogeneic immune effector cell.

10. An isolated nucleic acid fragment, wherein the nucleic acid fragment encodes an antibody or antigen-binding fragment thereof according to any one of claims 1 to 6, a multispecific antigen-binding molecule according to claim 7, or a chimeric antigen receptor according to claim 8.

11. A vector comprising the nucleic acid fragment described in claim 10.

12. A host cell comprising the vector described in claim 11, preferably the cell being a prokaryotic or eukaryotic cell, such as a bacterium (e.g., Escherichia coli), a fungus (e.g., yeast), an insect cell, or a mammalian cell (e.g., a CHO cell line or a 293T cell line).

13. A method for producing an antibody or antigen-binding fragment thereof according to any one of claims 1 to 6 or a multispecific antigen-binding molecule according to claim 7, the method comprising culturing the cells according to claim 12 and isolating the antibody, antigen-binding fragment, or multispecific antigen-binding molecule expressed by the cells.

14. A method for producing the aforementioned immune effector cells, the method comprising introducing a nucleic acid fragment encoding the CAR described in claim 8 into the immune effector cells, and optionally further comprising priming the immune effector cells to express the CAR described in claim 8.

15. A pharmaceutical composition comprising an antibody or antigen-binding fragment thereof according to any one of claims 1 to 6, a multispecific antigen-binding molecule according to claim 7, an immune effector cell according to claim 9, a nucleic acid fragment according to claim 10, a vector according to claim 11, or a product produced according to the method according to claim 13 or 14, wherein the pharmaceutical composition optionally further comprises a pharmaceutically acceptable carrier, diluent or adjuvant, and optionally further comprises an additional antitumor agent.

16. A method for treating a tumor or cancer, the method comprising administering to a subject an effective amount of an antibody or antigen-binding fragment thereof according to any one of claims 1 to 6, a multispecific antigen-binding molecule according to claim 7, an immune effector cell according to claim 9, a nucleic acid fragment according to claim 10, a vector according to claim 11, a product produced according to the method according to claim 13 or 14, or a pharmaceutical composition according to claim 15. Preferably, the tumor or cancer is a tumor or cancer expressing GPRC5D, preferably a B-cell lymphoma, and more preferably multiple myeloma (MM), in a method.

17. An antibody or antigen-binding fragment thereof according to any one of claims 1 to 6, a multispecific antigen-binding molecule according to claim 7, an immune effector cell according to claim 9, a nucleic acid fragment according to claim 10, a vector according to claim 11, a product produced according to the method according to claim 13 or 14, or a pharmaceutical composition according to claim 15, used in the manufacture of a drug for treating tumors or cancer, Preferably, the tumor or cancer is a tumor or cancer expressing GPRC5D, preferably a B-cell lymphoma, and more preferably multiple myeloma (MM), for use.

18. A kit comprising an antibody or antigen-binding fragment thereof according to any one of claims 1 to 6, a multispecific antigen-binding molecule according to claim 7, an immunoeffector cell according to claim 9, a nucleic acid fragment according to claim 10, a vector according to claim 11, a product produced according to the method according to claim 13 or 14, or a pharmaceutical composition according to claim 15.

19. A method for detecting GPRC5D expression in a biological sample, the method comprising contacting the biological sample with the antibody or antigen-binding fragment described in any one of claims 1 to 6 under conditions that allow for the formation of a complex between the antibody or antigen-binding fragment and GPRC5D, preferably the method further comprising detecting the formation of the complex and indicating the presence or expression level of GPRC5D in the sample.

20. Uses of the antibody or antigen-binding fragment thereof according to any one of claims 1 to 6 in the production of a GPRC5D detection reagent.

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