Novel Anti- cldn18.2 antibodies

Abstract

The present invention provides an anti-CLDN18.2 antibody or an antigen-binding fragment thereof, an isolated polynucleotide encoding the antibody or an antigen-binding fragment thereof, a pharmaceutical composition comprising the antibody or an antigen-binding fragment thereof, and uses thereof.

Description

Technical field

Prior Technology

Content of invention

Implementation

Claims

1. An anti-CLDN18.2 antibody or its antigen-binding fragment, comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises heavy chain HCDR1, HCDR2, and HCDR3 sequences, and the light chain variable region comprises light chain LCDR1, LCDR2, and LCDR3 sequences, wherein: a) HCDR1 contains the sequence of SEQ ID NO: 1, HCDR2 contains the sequence of SEQ ID NO: 19, and HCDR3 contains the sequence of SEQ ID NO: 21; LCDR1 contains the sequence of SEQ ID NO: 14, LCDR2 contains the sequence of SEQ ID NO: 16, and LCDR3 contains the sequence of SEQ ID NO: 18; b) HCDR1 contains the sequence of SEQ ID NO: 1, HCDR2 contains the sequence of SEQ ID NO: 3, and HCDR3 contains the sequence of SEQ ID NO: 5; LCDR1 contains the sequence of SEQ ID NO: 2, LCDR2 contains the sequence of SEQ ID NO: 4, and LCDR3 contains the sequence of SEQ ID NO: 6; c) HCDR1 contains the sequence of SEQ ID NO: 1, HCDR2 contains the sequence of SEQ ID NO: 7, and HCDR3 contains the sequence of SEQ ID NO: 5; LCDR1 contains the sequence of SEQ ID NO: 2, LCDR2 contains the sequence of SEQ ID NO: 4, and LCDR3 contains the sequence of SEQ ID NO:

18. d) The HCDR1 contains the sequence of SEQ ID NO: 1, the HCDR2 contains the sequence of SEQ ID NO: 9, and the HCDR3 contains the sequence of SEQ ID NO: 11; the LCDR1 contains the sequence of SEQ ID NO: 10, the LCDR2 contains the sequence of SEQ ID NO: 4, and the LCDR3 contains the sequence of SEQ ID NO: 6; e) The HCDR1 contains the sequence of SEQ ID NO: 13, the HCDR2 contains the sequence of SEQ ID NO: 15, and the HCDR3 contains the sequence of SEQ ID NO: 17; the LCDR1 contains the sequence of SEQ ID NO: 2, the LCDR2 contains the sequence of SEQ ID NO: 4, and the LCDR3 contains the sequence of SEQ ID NO: 12; or f) The HCDR1 contains the sequence of SEQ ID NO: 1, the HCDR2 contains the sequence of SEQ ID NO: 22, and the HCDR3 contains the sequence of SEQ ID NO: 5; the LCDR1 contains the sequence of SEQ ID NO: 20, the LCDR2 contains the sequence of SEQ ID NO: 8, the LCDR2 contains the sequence of SEQ ID NO: 9, and the LCDR3 contains the sequence of SEQ ID NO: 11; the LCDR1 contains the sequence of SEQ ID NO: 10, the LCDR2 contains the sequence of SEQ ID NO: 4, and the LCDR3 contains the sequence of SEQ ID NO: 6; The sequence of NO:4, and the sequence of SEQ ID NO:6 contained in the LCDR3.

2. The antibody or antigen-binding fragment thereof as claimed in claim 1, further comprising one or more heavy chains HFR1, HFR2, HFR3 and HFR4, and / or one or more light chains LFR1, LFR2, LFR3 and LFR4, wherein: HFR1 contains QVQLVQSGAEVKKPGASVKVSCKASGYX17FT (SEQ ID NO: 54), HFR2 contains WVX18QAPGQGLEWX19G (SEQ ID NO: 55), HFR3 contains RVTX20TIDKSTSTVYMELSSLRSEDTAVYYCAR (SEQ ID NO: 56), HFR4 contains WGQGTTVTVSS (SEQ ID NO: 57), LFR1 contains DIVMTQSPDSLAVSLGERATX21NC (SEQ ID NO: 58), LFR2 contains WYQQKPGQPPKLLIY (SEQ ID NO: 59), LFR3 contains GVPDRFX22GSGSGTDFTLTISSLQAEDVAVYYC (SEQ ID NO: 60), and LFR4 contains FGGGTKVEIK (SEQ ID NO: 60). NO: 61), where X17 is T or S, X18 is R or K, X19 is M or I, X20 is M or L, X21 is I or M, and X22 is S or T.

3. The antibody or its antigen-binding fragment as requested in claim 2, wherein: The HFR1 contains sequences selected from the following groups: SEQ ID NO: 62 and 63; the HFR2 contains sequences selected from the following groups: SEQ ID NO: 64 and 65; the HFR3 contains sequences selected from the following groups: SEQ ID NO: 66 and 67; the HFR4 contains a sequence of SEQ ID NO: 57; the LFR1 contains sequences selected from the following groups: SEQ ID NO: 68 and 69; the LFR2 contains a sequence of SEQ ID NO: 59; the LFR3 contains sequences selected from the following groups: SEQ ID NO: 70 and 71; and the LFR4 contains a sequence of SEQ ID NO:

61.

4. The antibody or antigen-binding fragment thereof as claimed in claim 1, wherein the variable region of the heavy chain comprises sequences selected from the following groups: SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45 and SEQ ID NO: 47; and wherein the variable region of the light chain comprises sequences selected from the following groups: SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46 and SEQ ID NO:

48.

5. The antibody or its antigen-binding fragment as requested in claim 1, wherein: The heavy chain variable region contains the sequence of SEQ ID NO: 25, and the light chain variable region contains the sequence of SEQ ID NO: 26; the heavy chain variable region contains the sequence of SEQ ID NO: 27, and the light chain variable region contains the sequence of SEQ ID NO: 28; the heavy chain variable region contains the sequence of SEQ ID NO: 29, and the light chain variable region contains the sequence of SEQ ID NO: 26 or 28; the heavy chain variable region contains the sequence of SEQ ID NO: 37, and the light chain variable region contains the sequence of SEQ ID NO: 38; the heavy chain variable region contains the sequence of SEQ ID NO: 39, and the light chain variable region contains the sequence of SEQ ID NO: 40; the heavy chain variable region contains the sequence of SEQ ID NO: 41, and the light chain variable region contains the sequence of SEQ ID NO: 42; the heavy chain variable region contains the sequence of SEQ ID NO: 43, and the light chain variable region contains the sequence of SEQ ID NO: 44; the heavy chain variable region contains the sequence of SEQ ID NO: 45, and the light chain variable region contains the sequence of SEQ ID NO:

26. The sequence of SEQ ID NO: 46; or the heavy chain variable region contains the sequence of SEQ ID NO: 47, and the light chain variable region contains the sequence of SEQ ID NO:

48.

6. The antibody or antigen-binding fragment thereof of claim 1 further comprises an immunoglobulin constant region, further comprises a human Ig constant region, or further comprises a human IgG constant region.

7. The antibody or antigen-binding fragment thereof as claimed in claim 6, wherein the constant region comprises a substitution for one or more amino acid residues in SEQ ID NO: 49, the substitution being selected from the group consisting of L235V, F243L, R292P, Y300L, P396L or any combination thereof.

8. The antibody or antigen-binding fragment thereof as claimed in claim 7, wherein the constant region contains the sequence of SEQ ID NO:

51.

9. The antibody or antigen-binding fragment thereof, as requested in item 1, is either humanized or non-fucosylated.

10. The antibody or its antigen-binding fragment, as requested in item 1, is a bifunctional antibody (diabody), Fab, Fab', F(ab')2, Fd, Fv fragment, disulfide-stabilized Fv fragment (dsFv), (dsFv)2, bispecific dsFv (dsFv-dsFv'), disulfide-stabilized bifunctional antibody (dsdiabody), single-chain antibody molecule (scFv), scFv dimer (bivalent bifunctional antibody), multispecific antibody, camelized single domain antibody, nano antibody, domain antibody, or bivalent domain antibody.

11. The antibody or its antigen-binding fragment as claimed in claim 1 is bispecific and can specifically bind to CLDN18.2 and the second antigen.

12. The antibody or antigen-binding fragment thereof as claimed in claim 11, wherein the second antigen is an immune-associated target, wherein the immune-associated target is selected from the following group: PD-L1, PD-L2, PD-1, CLTA-4, TIM-3, LAG3, CD160, 2B4, TGFβ, VISTA, BTLA, TIGIT, LAIR1, OX40, CD2, CD27, ICAM-1, NKG2C, SLAMF7, NKp80, CD160, B7-H3, LFA-1, ICOS, 4-1BB, GITR, CD30, CD40, BAFFR, HVEM, CD7, LIGHT, IL-2, IL-15, CD3, CD16, and CD83.

13. The antibody or antigen-binding fragment thereof as claimed in claim 11, wherein the second antigen comprises a tumor antigen, wherein the tumor antigen comprises CA-125, gangliosides G(D2), G(M2) and G(D3), CD20, CD52, CD33, Ep-CAM, CEA, bombesin-like peptides, PSA, HER2 / neu, epidermal growth factor receptor (EGFR), erbB2, erbB3 / HER3, erbB4, CD44v6, Ki-67, cancer-associated mucin, VEGF, VEGFR (e.g., VEGFR3), estrogen receptor, Lewis-Y antigen, TGFβ1, IGF-1 receptor, EGFα, c-Kit receptor, transferrin receptor, IL-2R or CO17-1A.

14. The antibody or antigen-binding fragment thereof of claim 1, which is linked to one or more conjugate moiety, wherein the conjugate moiety comprises a clearance-modifying agent, a chemotherapeutic agent, a toxin, a radioisotope, a lanthanide element, a luminescent label, a fluorescent label, an enzyme acceptor label, a DNA alkylating agent, a topoisomerase inhibitor, a tubulin binder (e.g., VCMMAE), or other anticancer drugs.

15. A pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof as claimed in any one of claims 1 to 14, and one or more pharmaceutically acceptable carriers.

16. An isolated polynucleotide encoding an antibody or antigen-binding fragment thereof as claimed in any one of claims 1 to 14.

17. A carrier comprising isolated polynucleotides as claimed in claim 16.

18. A host cell comprising the vector as claimed in claim 17.

19. A method of expressing an antibody or an antigen-binding fragment thereof as claimed in any one of claims 1 to 14, comprising culturing a host cell containing a vector comprising an isolated polynucleotide encoding an antibody or an antigen-binding fragment thereof as claimed in any one of claims 1 to 14, wherein the host cell is cultured under conditions expressing the vector.

20. Use of an antibody or antigen-binding fragment thereof as claimed in any one of claims 1 to 14 for the preparation of a pharmaceutical product, wherein the pharmaceutical product is intended to treat in an individual a disease or condition that may benefit from regulation of CLDN18.2 activity.

21. For the purposes of claim 20, wherein the disease or condition is cancer, optionally the cancer is cancer manifested as CLDN18.

2.

22. As used in claim 21, wherein the cancer exhibiting CLDN18.2 is a CLDN18.2 high-expressing cancer cell, a CLDN18.2 intermediate-expressing cancer cell, or a CLDN18.2 low-expressing cancer cell.

23. As claimed in claim 22, wherein the CLDN18.2 high-expressing cancer cells are CLDN18.2 at a level of at least 2+ intensity as determined by IHC and at least 70% of the cells in the IHC are positively stained; the CLDN18.2 intermediate-expressing cancer cells are CLDN18.2 at a level of at least 1+ intensity as determined by IHC and at least 40% but less than 70% of the cells in the IHC are positively stained; and the CLDN18.2 low-expressing cancer cells are CLDN18.2 at a level of at least 1+ intensity as determined by IHC and at least 0% but less than 40% of the cells in the IHC are positively stained.

24. For any of the uses in claims 21 to 23, wherein the cancer is gastric cancer, lung cancer, bronchial cancer, bone cancer, hepatobiliary cancer, pancreatic cancer, breast cancer, liver cancer, ovarian cancer, testicular cancer, kidney cancer, bladder cancer, head and neck cancer, spinal cancer, brain cancer, cervical cancer, uterine cancer, endometrial cancer, colon cancer, colorectal cancer, rectal cancer, anal cancer, esophageal cancer, gastrointestinal cancer, skin cancer, prostate cancer, pituitary cancer, stomach cancer, vaginal cancer, thyroid cancer, glioblastoma, astrocytoma, melanoma, myelodysplastic syndrome, sarcoma, teratoma, and adenocarcinoma.

25. As used in claim 20, wherein the individual is a person.

26. As claimed in claim 20, wherein the pharmaceutical product further comprises or is used in combination with a second therapeutic agent.

27. A kit comprising an antibody or an antigen-binding fragment thereof as claimed in any one of claims 1 to 14, and a second therapeutic agent.

28. A method for diagnosing a CLDN18.2-related disease or condition in an individual, comprising: a) contacting a sample obtained from the individual with an antibody or antigen-binding fragment thereof as claimed in any one of claims 1 to 14; b) determining the presence or amount of CLDN18.2 in the sample; and c) associating the presence or amount of CLDN18.2 with the presence or status of a CLDN18.2-related disease or condition in the individual.

29. A kit for detecting CLDN18.2, comprising an antibody or an antigen-binding fragment thereof as claimed in any one of claims 1 to 14.

30. A chimeric antigen receptor (CAR) comprising an antigen-binding domain, a transmembrane domain, a co-stimulatory signal transduction region, and a TCR signal transduction domain, wherein the antigen-binding domain is coupled to CLDN. 18.2 An antigen-binding fragment that specifically binds to and contains an antibody or an antigen-binding fragment thereof as claimed in any of claims 1 to 14.

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