Chimeric Antigen Receptors with MAGE-A4 Specificity and Uses Thereof

Patent metadata

Field	Value
Publication number	US-2024226163-A1
Application number	US-202218559009-A
Country	US
Kind code	A1
Filing date	May 3, 2022
Priority date	May 4, 2021
Publication date	Jul 11, 2024
Grant date	—

Abstract

MAGE-A4, or Melanoma-Associated Antigen A4, is a cancer-testis antigen (CTA) on the X chromosome. The present disclosure provides MAGE-A4-specific chimeric antigen receptors, cells expressing such chimeric antigen receptors, and MAGE-A4 specific isolated antibodies. In certain embodiments, engineered cells expressing the chimeric antigen receptors of the present disclosure are capable of inhibiting the growth of tumors expressing MAGE-A4. The engineered cells of the present disclosure are useful for the treatment of diseases and disorders in which an upregulated or induced MAGE-A4-targeted immune response is desired and/or therapeutically beneficial. For example, engineered cells expressing the MAGE-A4-specific chimeric antigen receptors of the present disclosure are useful for the treatment of various cancers.

First claim

What is claimed is:

1 . An antigen-binding protein that specifically binds an HLA-bound Melanoma-Associated Antigen A4 (MAGE-A4), wherein the antigen-binding protein comprises a light chain variable region (LCVR) and a heavy chain variable region (HCVR), wherein the LCVR comprises complementarity determining regions (CDRs) of a LCVR comprising the amino acid sequence of SEQ ID NO: 10 or SEQ ID NO: 115, and wherein the HCVR comprises CDRs of a HCVR comprising the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 83, or SEQ ID NO: 107.

2 . The antigen-binding protein of claim 1 , wherein the LCVR comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 10 or SEQ ID NO: 115.

3 . The antigen-binding protein of claim 1 or claim 2 , wherein the HCVR comprises an amino acid sequence having at least 95% identity to SEQ ID NO: 2, SEQ ID NO: 83, or SEQ ID NO: 107.

4 . The antigen-binding protein of any one of claims 1-3 , wherein said MAGE-A4-specific antigen-binding protein interacts with amino acids 286-294, or a portion thereof, of SEQ ID NO: 32.

5 . A MAGE-A4-specific chimeric antigen receptor (CAR) comprising from N-terminus to C-terminus: (a) an extracellular ligand-binding domain comprising an anti-MAGE-A4 single chain variable fragment (scFv) domain comprising a light chain variable region (LCVR) and a heavy chain variable region (HCVR); (b) a hinge; (c) a transmembrane domain; and (d) a cytoplasmic domain comprising a 4-1BB costimulatory domain or a CD28 costimulatory domain and a CD3zeta signaling domain, wherein the LCVR comprises complementarity determining regions (CDRs) of a LCVR comprising the amino acid sequence of SEQ ID NO: 10 or SEQ ID NO: 115, and CDRs of a HCVR comprising an amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 83 or SEQ ID NO: 107.

6 . The MAGE-A4 specific CAR of claim 5 , wherein said MAGE-A4-specific CAR comprises, from N-terminus to C-terminus: (a) the extracellular ligand-binding domain; (b) a hinge; (c) a transmembrane domain; and (d) a cytoplasmic domain comprising a costimulatory domain and a signaling domain.

7 . The MAGE-A4-specific CAR of claim 5 , wherein the anti-MAGE-A4 scFv domain comprises a first linker between the LCVR and the HCVR.

8 . The MAGE-A4-specific CAR of any one of claims 5-7 , further comprising a second linker between the extracellular ligand-binding domain and the hinge.

9 . The MAGE-A4-specific CAR of claim 8 , wherein the first linker comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-26, and wherein the second linker comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-26.

10 . The MAGE-A4-specific CAR of claim 9 , wherein the first linker comprises the amino acid sequence of SEQ ID NO: 25 and the second linker comprises the amino acid sequence of SEQ ID NO: 23.

11 . The MAGE-A4-specific CAR of any one of claims 5-10 , wherein the hinge, the transmembrane domain, or both, are from a CD8a polypeptide.

12 . The MAGE-A4-specific CAR of any one of claims 5-11 , wherein the costimulatory domain comprises a 4-1BB costimulatory domain.

13 . The MAGE-A4-specific CAR of any one of claims 5-11 , wherein the costimulatory domain comprises a CD28 costimulatory domain.

14 . The MAGE-A4-specific CAR of any one of claims 5-10, 12 and 13 , wherein the hinge, the transmembrane domain, or both, are from a CD28 polypeptide.

15 . The MAGE-A4-specific CAR of any one of claims 5-12 , wherein the hinge comprises the amino acid sequence of SEQ ID NO: 27.

16 . The MAGE-A4-specific CAR of any one of claims 5-12 , wherein the transmembrane domain comprises the amino acid sequence of SEQ ID NO: 28.

17 . The MAGE-A4-specific CAR of any one of claims 5-12 , wherein the 4-1BB costimulatory domain comprises the amino acid sequence of SEQ ID NO: 29.

18 . The MAGE-A4-specific CAR of any one of claims 5-10 and 12-14 , wherein the hinge comprises the amino acid sequence of SEQ ID NO: 34.

19 . The MAGE-A4-specific CAR of any one of claims 5-10, 12-14, and 18 , wherein the transmembrane domain comprises the amino acid sequence of SEQ ID NO: 36.

20 . The MAGE-A4-specific CAR of any one of claims 5-10, and 12-14, 18 and 19 , wherein the CD28 costimulatory domain comprises the amino acid sequence of SEQ ID NO: 38.

21 . The MAGE-A4-specific CAR of any one of claims 5-20 , wherein the signaling domain comprises a CD3zeta signaling domain.

22 . The MAGE-A4-specific CAR of claim 21 , wherein the CD3zeta signaling domain comprises the amino acid sequence of SEQ ID NO: 30.

23 . The antigen-binding protein of any one of claims 1-4 , wherein said antigen-binding protein is a MAGE-A4-specific antibody, or an antigen-binding fragment thereof.

24 . The antigen-binding protein of any one of claims 1-4 and 23 , or the MAGE-A4-specific CAR of any one of claims 5-22 , wherein said antigen-binding protein or MAGE-A4-specific CAR comprises three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) contained within a HCVR comprising the amino acid sequence set forth in SEQ ID NO: 2 or SEQ ID NO: 83.

25 . The antigen-binding protein or the MAGE-A4-specific CAR of claim 24 , wherein HCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 4, HCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 6, and HCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8.

26 . The antigen-binding protein or the MAGE-A4-specific CAR of claim 25 , wherein said HCVR comprises the amino acid sequence set forth in SEQ ID NO: 2.

27 . The antigen-binding protein or the MAGE-A4-specific CAR of claim 25 , wherein said HCVR comprises the amino acid sequence set forth in SEQ ID NO: 83.

28 . The antigen-binding protein of any one of claims 1-4 and 23 , or the MAGE-A4-specific CAR of any one of claims 5-22 , wherein said antigen-binding protein or MAGE-A4-specific CAR comprises three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) contained within a HCVR comprising the amino acid sequence set forth in SEQ ID NO: 107.

28 . The antigen-binding protein or the MAGE-A4-specific CAR of claim 28 , wherein HCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 109, HCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 111, and HCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 113.

29 . The antigen-binding protein or the MAGE-A4-specific CAR of claim 28 , wherein said HCVR comprises the amino acid sequence set forth in SEQ ID NO: 107.

30 . The antigen-binding protein of any one of claims 1-4 and 23 , the MAGE-A4-specific CAR of any one of claims 5-22 , or the antigen-binding protein or the MAGE-A4-specific CAR of any one of claims 24-29 , wherein said antigen-binding protein or MAGE-A4-specific CAR comprises three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained within a LCVR comprising the amino acid sequence set forth in SEQ ID NO: 10 or SEQ ID NO: 115.

31 . The antigen-binding protein or the MAGE-A4-specific CAR of claim 30 , wherein LCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 12, LCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 14, and LCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 16.

32 . The antigen-binding protein or the MAGE-A4-specific CAR of claim

Assignees

• Regeneron Pharma

Inventors

• Bray Kevin
• Delfino Frank
• Dilillo David

Classifications

• A61K40/11

  T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells · CPC title

• A61K40/31

  Chimeric antigen receptors [CAR] · CPC title

• A61K40/4268

  MAGE · CPC title

• A61K40/24

  Antigen-presenting cells [APC] · CPC title

• A61K35/17 Primary

  Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes (when activated by a specific antigen A61K39/00) · CPC title