Enhanced chimeric antigen receptors and uses thereof

Patent metadata

Field	Value
Publication number	US-2020317777-A1
Application number	US-202016911148-A
Country	US
Kind code	A1
Filing date	Jun 24, 2020
Priority date	Dec 29, 2017
Publication date	Oct 8, 2020
Grant date	—

Abstract

The presently disclosed subject matter provides methods and compositions for enhancing the immune response toward cancers and pathogens. It relates to novel designs of chimeric antigen receptors (CARs) and engineered immunoresponsive cells comprising the same. The engineered immunoresponsive cells comprising the novel CARs are antigen-directed and have extended persistence without compromising function.

First claim

What is claimed is:

1 . A chimeric antigen receptor (CAR) comprising an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain comprising a modified CD3ζ polypeptide, wherein the modified CD3ζ polypeptide lacks all or part of immunoreceptor tyrosine-based activation motifs (ITAMs), wherein the ITAMs are ITAM1, ITAM2, and ITAM3.

2 . The CAR of claim 1 , wherein the modified CD3ζ polypeptide lacks ITAM2 or a portion thereof.

3 . The CAR of claim 2 , wherein the modified CD3ζ polypeptide further lacks ITAM3 or a portion thereof.

4 . The CAR of claim 2 , wherein the modified CD3ζ polypeptide further lacks ITAM1 or a portion thereof.

5 . The CAR of claim 1 , wherein the modified CD3ζ polypeptide lacks ITAM1 or a portion thereof.

6 . The CAR of claim 5 , wherein the modified CD3ζ polypeptide further lacks ITAM3 or a portion thereof.

7 . The CAR of claim 1 , wherein the modified CD3ζ polypeptide lacks ITAM3 or a portion thereof.

8 . The CAR of claim 1 , wherein the modified CD3ζ polypeptide comprises a deletion of ITAM2 or a portion thereof.

9 . The CAR of claim 8 , wherein the modified CD3ζ polypeptide further comprises a deletion of ITAM3 or a portion thereof.

10 . The CAR of claim 8 , wherein the modified CD3ζ polypeptide further comprises a deletion of ITAM1 or a portion thereof.

11 . The CAR of claim 1 , wherein the modified CD3ζ polypeptide comprises a deletion of ITAM1 or a portion thereof.

12 . The CAR of claim 11 , wherein the modified CD3ζ polypeptide further comprises a deletion of ITAM3 or a portion thereof.

13 . The CAR of claim 1 , wherein the modified CD3ζ polypeptide comprises a deletion of ITAM3 or a portion thereof.

14 . The CAR of claim 1 , wherein the modified CD3ζ polypeptide lacks all or part of basic-rich stretch (BRS) regions, wherein the BRS regions are BRS1, BRS2, and BRS3.

15 . The CAR of claim 14 , wherein the modified CD3ζ polypeptide lacks BRS2 or a portion thereof.

16 . The CAR of claim 15 , wherein the modified CD3ζ polypeptide further lacks BRS3 or a portion thereof.

17 . The CAR of claim 16 , wherein the modified CD3ζ polypeptide further lacks BRS1 or a portion thereof.

18 . The CAR of claim 14 , wherein the modified CD3ζ polypeptide lacks BRS1 or a portion thereof.

19 . The CAR of claim 18 , wherein the modified CD3ζ polypeptide further lacks BRS3 or a portion thereof.

20 . The CAR of claim 14 , wherein the modified CD3ζ polypeptide lacks BRS3 or a portion thereof.

21 . The CAR of claim 14 , wherein the modified CD3ζ polypeptide lacks BRS1 or portion thereof, BRS2 or portion thereof, and BRS3 or a portion thereof.

22 . The CAR of claim 1 , wherein the modified CD3ζ polypeptide lacks ITAM2, ITAM3, BRS2, and BRS3.

23 . The CAR of claim 14 , wherein the modified CD3ζ polypeptide comprises a deletion of BRS2 or a portion thereof.

24 . The CAR of claim 23 , wherein the modified CD3ζ polypeptide further comprises a deletion of BRS3 or a portion thereof.

25 . The CAR of claim 24 , wherein the modified CD3ζ polypeptide further comprises a deletion of BRS1 or a portion thereof.

26 . The CAR of claim 14 , wherein the modified CD3ζ polypeptide comprises a deletion of BRS1 or a portion thereof.

27 . The CAR of claim 26 , wherein the modified CD3ζ polypeptide further comprises a deletion of BRS3 or a portion thereof.

28 . The CAR of claim 14 , wherein the modified CD3ζ polypeptide comprises a deletion of BRS3 or a portion thereof.

29 . The CAR of claim 14 , wherein the modified CD3ζ polypeptide comprises a deletion of BRS1 or portion thereof, BRS2 or portion thereof, and BRS3 or a portion thereof.

30 . The CAR of claim 1 , wherein the modified CD3ζ polypeptide comprises a deletion of ITAM2, ITAM3, BRS2, and BRS3.

31 . The CAR of claim 1 , wherein the CAR comprises the amino acid sequence set forth in SEQ ID NO: 45 or SEQ ID NO: 47.

32 . A chimeric antigen receptor (CAR) comprising an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain comprising a modified CD3ζ polypeptide, wherein the modified CD3ζ polypeptide lacks all or part of basic-rich stretch (BRS) regions, wherein the BRS regions are BRS1, BRS2, and BRS3.

33 . The CAR of claim 32 , wherein the modified CD3ζ polypeptide lacks BRS2 or a portion thereof.

34 . The CAR of claim 33 , wherein the modified CD3ζ polypeptide further lacks BRS3 or a portion thereof.

35 . The CAR of claim 34 , wherein the modified CD3ζ polypeptide further lacks BRS1 or a portion thereof.

36 . The CAR of claim 35 , wherein the modified CD3ζ polypeptide lacks BRS1 or a portion thereof.

37 . The CAR of claim 36 , wherein the modified CD3ζ polypeptide further lacks BRS3 or a portion thereof.

38 . The CAR of claim 37 , wherein the modified CD3ζ polypeptide lacks BRS3 or a portion thereof.

39 . The CAR of claim 32 , wherein the modified CD3ζ polypeptide lacks BRS1 or portion thereof, BRS2 or portion thereof, and BRS3 or a portion thereof.

40 . A chimeric antigen receptor (CAR) comprising an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain comprising a modified CD3ζ polypeptide, wherein the modified CD3ζ polypeptide comprises a BRS variant selected from a BRS1 variant, a BRS2 variant, and a BRS3 variant, wherein the BRS variant comprises one or more loss-of-function mutations.

41 . The CAR of claim 1 , further comprising a hinge/spacer region, wherein the hinge/spacer region comprises a CD8 polypeptide, a CD28 polypeptide, a CD3ζ polypeptide, a CD4 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, a CD166 polypeptide, a CD166 polypeptide, a CD8a polypeptide, a CD8b polypeptide, an ICOS polypeptide, an ICAM-1 polypeptide, a CTLA-4 polypeptide, a CD27 polypeptide, a CD40/My88 peptide, a NKGD2 peptide, or a combination thereof.

42 . The CAR of claim 41 , wherein the hinge/spacer region comprises a CD166 polypeptide.

43 . The CAR of claim 42 , wherein the CD166 polypeptide has amino acids 489 to 527 of SEQ ID NO: 3.

44 . A chimeric antigen receptor (CAR) comprising an extracellular antigen-binding domain, a hinge/spacer region, a transmembrane domain, and an intracellular signaling domain comprising a modified CD3ζ polypeptide,
wherein the modified CD3ζ polypeptide comprises one or more ITAM variant comprising one or more loss-of-function mutations, wherein each of the one or more ITAM variant is independently selected from the group consisting of an ITAM1 variant, an ITAM2 variant, and an ITAM3 variant, and
wherein the hinge/spacer region comprises a CD8 polypeptide, a CD28 polypeptide, a CD3ζ polypeptide, a CD4 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, a CD166 polypeptide, a CD166 polypeptide, a CD8a polypeptide, a CD8b polypeptide, an ICOS polypeptide, an ICAM-1 polypeptide, a CTLA-4 polypeptide, a CD27 polypeptide, a CD40/My88 peptide, a NKGD2 peptide, or a combination thereof.

45 . The method of claim 44 , wherein the modified CD3ζ polypept

Assignees

• Memorial Sloan Kettering Cancer Center

Inventors

• Sadelain Michel
• Feucht Judith
• Hamieh Mohamad
• Sun Jie
• Mansilla-Soto Jorge A

Classifications

• A61K2239/10

  characterized by the structure of the chimeric antigen receptor [CAR] · CPC title

• A61P35/02

  specific for leukemia · CPC title

• C07K14/7051

  T-cell receptor (TcR)-CD3 complex · CPC title

• A61K40/31 Primary

  Chimeric antigen receptors [CAR] · CPC title

• C07K2319/03

  containing a transmembrane segment · CPC title