IL298326A

IL298326A - Anti-her2 antibody or antigen-binding fragment thereof, and chimeric antigen receptor comprising same

Info

Publication number: IL298326A
Application number: IL298326A
Authority: IL
Original assignee: Gc Cell Corp; Abclon Inc
Priority date: 2020-05-22
Filing date: 2021-05-21
Publication date: 2023-01-01
Also published as: EP4153635A1; WO2021235894A1; EP4153635A4; KR20230024911A; CA3184449A1

Description

1 WO 2021/235894 PCT/KR2021/006361 Description Title of Invention: Anti-HER2 Antibody or Antigen-binding Fragment thereof, and Chimeric Antigen Receptor Comprising Same Technical Field [1] The instant application includes a Sequence Listing which has been submitted elec- tronically in ASCII and is hereby incorporated by reference in its entirety. Said ASCII copy, created on May 5, 2020, is named 4411-0210002_Sequences_ST25.txt and is 151,877 bytes in size.[2] The research was conducted under the support of the Ministry of Trade, Industry and Energy of Korea with the project number 1415118385. The R&D management agency of the project is the Korea Institute for Advancement of Technology, the R&D project title is "Global innovation technology alliance", and the research title is "Development of global antibody drug based on novel epitope screening platform technology". The research was conducted by AbClon Inc. from November 1, 2011 until October 31, 2014.[3] This application claims the priority of Korean Patent Application No. 10-2017-0151841 filed on November 14, 2017 in the Korean Intellectual Property Office, the disclosure of which is incorporated herein by reference in its entirety.[4] The present disclosure relates to a novel anti-HER2 antibody or an antigen-binding fragment thereof, a chimeric antigen receptor including the same, and uses thereof.[5] Background Art [6] The Her2/neu (ErbB2) gene encodes a 185-kDa transmembrane glycoprotein that belongs to the family of epidermal growth factor receptors (EGFRs). The Her2 protein is composed of an extracellular domain consisting of 620 amino acid residues, a trans- membrane domain 23 amino acid residues, and an intracellular domain with tyrosine kinase activity, consisting of 490 amino acid residues (Akiyama T, et al., Science, 2(4758): 1644-1646(1986)).[7] Anti-HER2 antibodies with various characteristics have been described: Tagliabue et al., Int. J. Cancer 47: 933-937 (1991); McKenzie et al., Oncogene 4: 543-548 (1989); Maier et al., Cancer Res. 51: 5361-5369 (1991); Bacus et al., Molecular Car- cinogenesis 3: 350-362 (1990); Stancovski et al., PNAS USA 88: 8691-8695 (1991); Bacus et al., Cancer Research 52: 2580-2589 (1992); Xu et al., Int. J. Cancer 53: 401-408 (1993); WO94/00136; Kasprzyk et al., Cancer Research 52: 2771-27(1992); Hancock et al., Cancer Res. 51: 4575-4580 (1991); Shawver et al., Cancer Res. 2 WO 2021/235894 PCT/KR2021/006361 54: 1367-1373 (1994); Arteaga et al., Cancer Res. 54: 3758-3765 (1994); Harwerth et al., J. Biol. Chem. 267: 15160-15167 (1992); US Patent No. 5,783,186; Kao et al., US Patent Application Publication No. 2009/0285837 (2009); Ross et al., The Oncologist 8: 307-325 (2003); and Klapper et al., Oncogene 14: 2099-2109 (1997).[8] The most commercially successful anti-HER2 antibody is trastuzumab antibody (commercially available as HerceptinTM, US Patent No. 5,821,337) and many re- searches have been conducted thereon: Sapino, A., et al., Annals of Oncology (2007) 18: 1963-1968; Bussolati, G, et al., British Journal of Cancer (2005) 92, 1261-1267; and Glazyrin A, et al., J Histology & Cytochemistry (2007) 55 (1): 25-33.[9] Although the trastuzumab antibody has been commercially successful, use of the trastuzumab antibody for therapeutic purposes is limited because there are various cancer cells which have non-reactivity (or resistance) to the antibody or have reduced sensitivity. Accordingly, there have been attempts to resolve the therapeutic problem of the antibody.[10] For example, US Patent No. 7,674,460 discloses a method for increasing the HERsensitivity of cancer cells using an HER2 antagonist such as the trastuzumab antibody and a PC cell-derived growth factor (PCDGF) antagonist. WO 2011/127297 discloses a method for inhibiting the proliferation of trastuzumab-resistant tumor cells using a combination of a FoxMl inhibitor and the trastuzumab antibody.[11] US Patent Application Publication No. 2010-0183604 discloses a method for treating trastuzumab-resistant cancer using a cofilin inhibitor, a PAK1 inhibitor, a LIMK inhibitor, an RHO inhibitor, a ROCK1 inhibitor or a ROCK2 inhibitor.[12] Disclosure of Invention Technical Problem [13] The inventors of the present disclosure have made efforts to develop a novel antibody which is capable of preventing or treating cancer (particularly, breast cancer and gastric cancer), exhibits better killing ability (or proliferation-inhibiting ability) for cancer cells which have non-reactivity (or resistance) to the trastuzumab antibody or have reduced sensitivity, and is capable of preventing or treating cancer with improved anticancer activity when co-administered with the trastuzumab antibody as compared to single administration of trastuzumab. As a result, they have developed a novel antibody which exhibits better killing ability for HER2-overexpressed cancer cells on which the trastuzumab antibody hardly acts, or exhibits improved anticancer activity when co-administered with the trastuzumab antibody, and have completed the present disclosure.[14] 3 WO 2021/235894 PCT/KR2021/006361

Claims

1.WO 2021/235894 PCT/KR2021/006361

2.Claims [Claim 1] An antibody against HER2 (human epidermal growth factor receptor 2) comprising any one of (a) to (e), or an antigen-binding fragment thereof:(a) a heavy chain variable region comprising CDRH1 of SEQ ID NO: 1, CDRH2 of SEQ ID NO: 2 and CDRH3 of SEQ ID NO: 3, and a light chain variable region comprising CDRL1 of SEQ ID NO: 4, CDRL2 of SEQ ID NO: 5 and CDRL3 of SEQ ID NO: 6;(b) a heavy chain variable region comprising CDRH1 of SEQ ID NO: 7, CDRH2 of SEQ ID NO: 8 and CDRH3 of SEQ ID NO: 9, 71 or 72, and a light chain variable region comprising CDRL1 of SEQ ID NO: 10, CDRL2 of SEQ ID NO: 11 and CDRL3 of SEQ ID NO: 12, 73 or 74;(c) a heavy chain variable region comprising CDRH1 of SEQ ID NO: 13, CDRH2 of SEQ ID NO: 14 and CDRH3 of SEQ ID NO: 15, and a light chain variable region comprising CDRL1 of SEQ ID NO: 16, CDRL2 of SEQ ID NO: 17 and CDRL3 of SEQ ID NO: 18;(d) a heavy chain variable region comprising CDRH1 of SEQ ID NO: 19, CDRH2 of SEQ ID NO: 20 and CDRH3 of SEQ ID NO: 21, and a light chain variable region comprising CDRL1 of SEQ ID NO: 22, CDRL2 of SEQ ID NO: 23 and CDRL3 of SEQ ID NO: 24; and(e) a heavy chain variable region comprising CDRH1 of SEQ ID NO: 25, CDRH2 of SEQ ID NO: 26 and CDRH3 of SEQ ID NO: 27, and a light chain variable region comprising CDRL1 of SEQ ID NO: 28, CDRL2 of SEQ ID NO: 29 and CDRL3 of SEQ ID NO: 30.[Claim 2] The antibody or the antigen-binding fragment thereof according to claim 1, whereinthe heavy chain variable region of (a) comprises an amino acid sequence of SEQ ID NO: 31 or 75;the heavy chain variable region of (b) comprises an amino acidsequence of SEQ ID NO: 39, 83, 87, 95 or 103;the heavy chain variable region of (c) comprises an amino acid sequence of SEQ ID NO: 47;the heavy chain variable region of (d) comprises an amino acid sequence of SEQ ID NO: 55; andthe heavy chain variable region of (e) comprises an amino acid sequence of SEQ ID NO: 63 or 79. 49 WO 2021/235894 PCT/KR2021/006361

3.[Claim 3] The antibody or the antigen-binding fragment thereof according toclaim 1, whereinthe light chain variable region of (a) comprises an amino acid sequence of SEQ ID NO: 35 or 77;the light chain variable region of (b) comprises an amino acid sequence of SEQ ID NO: 43, 85, 91, 99 or 107;the light chain variable region of (c) comprises an amino acid sequence of SEQ ID NO: 51;the light chain variable region of (d) comprises an amino acid sequence of SEQ ID NO: 59; andthe light chain variable region of (e) comprises an amino acid sequence of SEQ ID NO: 67 or 81.

4.[Claim 4] The antibody or the antigen-binding fragment thereof according toclaim 1, whereinthe antibody or the antigen-binding fragment thereof comprising (a) comprises a heavy chain comprising an amino acid sequence of SEQ ID NO: 33;the antibody or the antigen-binding fragment thereof comprising (b) comprises a heavy chain comprising an amino acid sequence of SEQ ID NO: 41, 89, 97 or 105;the antibody or the antigen-binding fragment thereof comprising (c) comprises a heavy chain comprising an amino acid sequence of SEQ ID NO: 49;the antibody or the antigen-binding fragment thereof comprising (d) comprises a heavy chain comprising an amino acid sequence of SEQ ID NO: 57; andthe antibody or the antigen-binding fragment thereof comprising (e) comprises a heavy chain comprising an amino acid sequence of SEQ ID NO: 65.

5.[Claim 5] The antibody or the antigen-binding fragment thereof according toclaim 1, whereinthe antibody or the antigen-binding fragment thereof comprising (a) comprises a light chain comprising an amino acid sequence of SEQ ID NO: 37;the antibody or the antigen-binding fragment thereof comprising (b) comprises a light chain comprising an amino acid sequence of SEQ ID NO: 45, 93, 101 or 109;the antibody or the antigen-binding fragment thereof comprising (c) 50 WO 2021/235894 PCT/KR2021/006361 comprises a light chain comprising an amino acid sequence of SEQ ID NO: 53;the antibody or the antigen-binding fragment thereof comprising (d) comprises a light chain comprising an amino acid sequence of SEQ ID NO: 61; andthe antibody or the antigen-binding fragment thereof comprising (e) comprises a light chain comprising an amino acid sequence of SEQ ID NO: 69.

6.[Claim 6] A fusion protein comprising the antibody or the antigen-binding fragment thereof according to any of claims 1 to 5.

7.[Claim 7] A chimeric antigen receptor polypeptide comprising:(a) an HER2-binding domain;(b) a transmembrane domain (TM);(c) a costimulatory domain; and(d) an intracellular signaling domain (ICD).

8.[Claim 8] The chimeric antigen receptor polypeptide according to claim 7, wherein the HER2-binding domain comprises the antibody or the antigen-binding fragment thereof according to any of claims 1 to 5.

9.[Claim 9] The chimeric antigen receptor polypeptide according to claim 7, wherein the transmembrane domain is a transmembrane domain of a protein selected from a group consisting of T-cell receptor alpha, beta or zeta chain, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137 and CD154.

10.[Claim 10] The chimeric antigen receptor polypeptide according to claim 7, wherein the costimulatory domain is a functional signaling domain obtained from a protein selected from a group consisting of MHC class I molecule, TNF receptor protein, immunoglobulin-like protein, cytokine receptor, integrin, signaling lymphocytic activation molecule (SLAM), activating NK cell receptor, BTLA (B- and T-lymphocyte at- tenuator), Toll-like ligand receptor, 0X40, CD2, CD7, CD27, CD28, CD30, CD40, CDS, ICAM-1, LFA-1 (CDlla/CD18), 4-1BB (CD137), B7-H3, CDS, ICAM-1, ICOS (CD278), GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD 19, CD4, CD8alpha, CD8 beta, IL2R beta, IL2R gamma, IL7R alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CDlld, ITGAE, CD103, ITGAL, CDlla, LFA-1, ITGAM, CDllb, ITGAX, CDllc, ITGB1, CD29, 51 WO 2021/235894 PCT/KR2021/006361 ITGB2, CD 18, LFA-1, ITGB7, NKG2D, NKG2C, TNFR2, TRANCE/ RANKE, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD(Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD160 (BY55), PSGL1, CD 100 (SEMA4D), CD69, SLAMF6 (NTB-A, LylO8), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, EAT, GADS, SLP-76, PAG/Cbp, CD19a, and a ligand binding specifically to CD83.

11.[Claim 11] The chimeric antigen receptor polypeptide according to claim 7, wherein the intracellular signaling domain comprises a functional signaling domain of 4-IBB, CD28, 0X40 or CD3 zeta, or a com- bination thereof.

12.[Claim 12] A nucleic acid molecule encoding the anti-HER2 antibody or the antigen-binding fragment thereof according to claim 1.

13.[Claim 13] A nucleic acid molecule encoding the chimeric antigen receptor polypeptide according to claim 7.

14.[Claim 14] A recombinant vector comprising the nucleic acid molecule according to claim 12 or 13.

15.[Claim 15] A host cell transformed with the recombinant vector according to claim 4

16.[Claim 16]I-r.An effector cell expressing the chimeric antigen receptor polypeptide according to claim 7.

17.[Claim 17] The effector cell according to claim 16, wherein the effector cell is selected from a group consisting of a dendritic cell, a killer dendritic cell, a mast cell, a natural killer cell, a B lymphocyte, a T lymphocyte, a macrophage and precursor cells thereof.

18.[Claim 18] The effector cell according to claim 17, wherein the T lymphocyte is selected from a group consisting of an inflammatory T lymphocyte, a cytotoxic T lymphocyte, a regulatory T lymphocyte or a helper T lymphocyte.

19.[Claim 19] A pharmaceutical composition for preventing or treating cancer, comprising: (a) a pharmaceutically effective amount of the anti-HERantibody or the antigen-binding fragment thereof according to any of claims 1 to 3; and (b) a pharmaceutically acceptable carrier.

20.[Claim 20] A pharmaceutical composition for treating cancer, comprising the effector cell expressing the chimeric antigen receptor polypeptide according to claim 16.

21.[Claim 21] The composition according to claim 19 or 20, wherein the cancer is breast cancer, ovarian cancer, gastric cancer, lung cancer, liver cancer, 52 WO 2021/235894 PCT/KR2021/006361 bronchial cancer, nasopharyngeal cancer, laryngeal cancer, pancreatic cancer, bladder cancer, colorectal cancer, colon cancer, cervical cancer, brain cancer, prostate cancer, bone cancer, head and neck cancer, skin cancer, thyroid cancer, parathyroid cancer or ureteral cancer.

22.[Claim 22] The composition according to claim 19 or 20, wherein the pharma- ceutical composition further comprises the trastuzumab antibody.

23.[Claim 23] A kit for diagnosing cancer, comprising the anti-HER2 antibody or the antigen-binding fragment thereof according to any of claims 1 to 5.

24.[Claim 24] A chimeric antigen receptor comprising an extracellular domain that binds Her2, wherein the extracellular domain comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 1(hz39D2 (VL-GS linker-VH)).

25.[Claim 25] The chimeric antigen receptor of claim 24, wherein the extracellular domain comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 113 (hz39D2 (VL-GS linker-VH)).

26.[Claim 26] The chimeric antigen receptor of claim 24, wherein the extracellular domain comprises an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 113 (hz39D2 (VL-GS linker-VH)).

27.[Claim 27] The chimeric antigen receptor of claim 24, wherein the extracellular domain comprises SEQ ID NO: 113 (hz39D2 (VL-GS linker-VH)).

28.[Claim 28] The chimeric antigen receptor of claim 24, further comprising an extracellular signaling domain linked to the extracellular domain; a hinge domain linked to the extracellular domain;a transmembrane domain linked to the hinge domain; and an intracellular stimulatory signal linked to the hinge domain.

29.[Claim 29] The chimeric antigen receptor of claim 28, wherein the extracellular signaling domain comprises an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 111 (CD8a signal peptide).

30.[Claim 30] The chimeric antigen receptor of claim 29, wherein the extracellular signaling domain comprises SEQ ID NO: 111 (CD8a signal peptide).

31.[Claim 31] The chimeric antigen receptor of claim 30, wherein the hinge domain comprises an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 115 (CD8a hinge).

32.[Claim 32] The chimeric antigen receptor of claim 31, wherein the hinge domain comprises SEQ ID NO: 115 (CD8a hinge).

33.[Claim 33] The chimeric antigen receptor of claim 32, wherein the transmembrane domain comprises an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 117 (CD8a TM) or SEQ ID NO: 53 WO 2021/235894 PCT/KR2021/006361 119(CD28 TM).

34.[Claim 34] The chimeric antigen receptor of claim 33, wherein the transmembranedomain comprises SEQ ID NO: 117 CD8a TM) or SEQ ID NO: 1(CD28 TM).

35.[Claim 35] The chimeric antigen receptor of claim 34, wherein the intracellularstimulatory signal comprises an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 121 (CD3-؛).

36.[Claim 36] The chimeric antigen receptor of claim 35, wherein the intracellularstimulatory signal comprises SEQ ID NO: 121 (CD3-؛).

37.[Claim 37] The chimeric antigen receptor of claim 36, further comprising a secondintracellular stimulatory signal, wherein the second intracellular stimulatory signal comprises an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 123 (4-IBB) or SEQ ID NO: 125 (CD28).

38.[Claim 38] The chimeric antigen receptor of claim 37, wherein the second intra-cellular stimulatory signal comprises SEQ ID NO: 123 (4-IBB) or SEQ ID NO: 125 (CD28).

39.[Claim 39] The chimeric antigen receptor of claim 38, further comprising a thirdintracellular stimulatory signal, wherein the third intracellular stimulatory signal comprises an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 127 (OX40L).

40.[Claim 40] The chimeric antigen receptor of claim 39, wherein the third intra-cellular stimulatory signal comprises SEQ ID NO: 127 (OX40L).

41.[Claim 41] The chimeric antigen receptor of claim 28, wherein the chimericantigen receptor comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 129 (Clone 2), SEQ ID NO: 1(Clone 3), SEQ ID NO: 133 (Clone 6), or SEQ ID NO: 135 (Clone 14).

42.[Claim 42] The chimeric antigen receptor of claim 28, wherein the chimericantigen receptor comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 129 (Clone 2), SEQ ID NO: 1(Clone 3), SEQ ID NO: 133 (Clone 6), or SEQ ID NO: 135 (Clone 14).

43.[Claim 43] The chimeric antigen receptor of claim 28, wherein the chimericantigen receptor comprises an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 129 (Clone 2), SEQ ID NO: 1(Clone 3), SEQ ID NO: 133 (Clone 6), or SEQ ID NO: 135 (Clone 14).

44.[Claim 44] The chimeric antigen receptor of claim 28, wherein the chimericantigen receptor comprises SEQ ID NO: 129 (Clone 2), SEQ ID NO: 131 (Clone 3), SEQ ID NO: 133 (Clone 6), or SEQ ID NO: 135 (Clone 54 WO 2021/235894 PCT/KR2021/006361 14).

45.[Claim 45] A nucleic acid molecule encoding the chimeric antigen receptor of claim 24.

46.[Claim 46]

47.[Claim 47]

48.[Claim 48] A vector comprising the nucleic acid molecule of claim 45.An immune cell expressing the chimeric antigen receptor of claim 24.The immune cell of claim 47, wherein the immune cell is a natural killer cell.

49.[Claim 49] A pharmaceutical composition comprising the immune cell of claim and a pharmaceutically acceptable carrier.

50.[Claim 50] A method for treating cancer comprising administering to a subject in need thereof a therapeutically effective amount of the pharmaceutical composition of claim 49.

51.[Claim 51] The method of claim 50, wherein the cancer is selected from the group consisting of breast cancer, ovarian cancer, gastric cancer, lung cancer, liver cancer, bronchial cancer, nasopharyngeal cancer, laryngeal cancer, pancreatic cancer, bladder cancer, colorectal cancer, colon cancer, cervical cancer, brain cancer, prostate cancer, bone cancer, head and neck cancer, skin cancer, thyroid cancer, parathyroid cancer and ureteral cancer.

52.[Claim 52] The chimeric antigen receptor of claim 7, wherein the intracellular signaling domain comprises a functional signaling domain of 0Xligand.