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US-20260124303-A1 - FIBROBLAST ACTIVATION PROTEIN (FAP) CAR-INVARIANT NATURAL KILLER T CELLS AND USES THEREOF

US20260124303A1US 20260124303 A1US20260124303 A1US 20260124303A1US-20260124303-A1

Abstract

The present disclosure, at least in part, is based on the discovery of novel anti-Fibroblast activation protein (FAP) antibodies or antigen binding fragments thereof, and genetically modified cells (e.g., iNKT cells) expressing chimeric antigen receptors comprising the anti-FAP antibody or antigen binding fragment thereof demonstrate improved properties, including increased binding to FAP, killing of FAP-expressing cancer cells in vitro and in vivo; and enhanced persistent in a subject receiving the therapy.

Inventors

  • Xavier MICHELET
  • Vignesh Venkatraman
  • Paul lbbett
  • Eleni CHANTZOURA
  • Olivier LE TONQUEZE
  • Efrat ALTMAN-SHARONI
  • Martyna POPIS
  • Magdalena NIEDZIELSKA
  • Marc van Dijk
  • Dhan CHAND
  • Olga Ignatovich

Assignees

  • MINK THERAPEUTICS, INC.
  • AGENUS INC.

Dates

Publication Date
20260507
Application Date
20231004

Claims (20)

  1. 1 . An invariant natural killer T (iNKT) cell comprising a chimeric antigen receptor (CAR) that specifically binds fibroblast activation protein (FAP), wherein the chimeric antigen receptor comprises: (a) a CDRH1 having the amino acid sequence of SEQ ID NO: 1, a CDRH2 having the amino acid sequence of SEQ ID NO: 2, a CDRH3 having the amino acid sequence of SEQ ID NO: 3, a CDRL1 having the amino acid sequence of SEQ ID NO: 4, a CDRL2 having the amino acid sequence of SEQ ID NO: 5, and a CDRL3 having the amino acid sequence of SEQ ID NO: 6; (b) a CDRH1 having the amino acid sequence of SEQ ID NO: 10, a CDRH2 having the amino acid sequence of SEQ ID NO: 11, a CDRH3 having the amino acid sequence of SEQ ID NO: 12, a CDRL1 having the amino acid sequence of SEQ ID NO: 4, a CDRL2 having the amino acid sequence of SEQ ID NO: 5, and a CDRL3 having the amino acid sequence of SEQ ID NO: 13; (c) a CDRH1 having the amino acid sequence of SEQ ID NO: 40, a CDRH2 having the amino acid sequence of SEQ ID NO: 41, a CDRH3 having the amino acid sequence of SEQ ID NO: 42, a CDRL1 having the amino acid sequence of SEQ ID NO: 4, a CDRL2 having the amino acid sequence of SEQ ID NO: 5, and a CDRL3 having the amino acid sequence of SEQ ID NO: 43; (d) a CDRH1 having the amino acid sequence of SEQ ID NO: 10, a CDRH2 having the amino acid sequence of SEQ ID NO: 11, a CDRH3 having the amino acid sequence of SEQ ID NO: 42, a CDRL1 having the amino acid sequence of SEQ ID NO: 4, a CDRL2 having the amino acid sequence of SEQ ID NO: 5, and a CDRL3 having the amino acid sequence of SEQ ID NO: 43; or (e) a CDRH1 having the amino acid sequence of SEQ ID NO: 10, a CDRH2 having the amino acid sequence of SEQ ID NO: 11, a CDRH3 having the amino acid sequence of SEQ ID NO: 54, a CDRL1 having the amino acid sequence of SEQ ID NO: 4, a CDRL2 having the amino acid sequence of SEQ ID NO: 5, and a CDRL3 having the amino acid sequence of SEQ ID NO: 55.
  2. 2 . The iNKT cell of claim 1 , wherein the CAR comprises a CDRH1 having the amino acid sequence of SEQ ID NO: 10, a CDRH2 having the amino acid sequence of SEQ ID NO: 11, a CDRH3 having the amino acid sequence of SEQ ID NO: 42, a CDRL1 having the amino acid sequence of SEQ ID NO: 4, a CDRL2 having the amino acid sequence of SEQ ID NO: 5, and a CDRL3 having the amino acid sequence of SEQ ID NO: 43.
  3. 3 . The iNKT cell of claim 1 or 2 , wherein the CAR comprises: (a) a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 7 and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 8; (b) a VH comprising the amino acid sequence of SEQ ID NO: 14 and a VL comprising the amino acid sequence of SEQ ID NO: 15; (c) a VH comprising the amino acid sequence of SEQ ID NO: 44 and a VL comprising the amino acid sequence of SEQ ID NO: 45; (d) a VH comprising the amino acid sequence of SEQ ID NO: 47 and a VL comprising the amino acid sequence of SEQ ID NO: 48; or (e) a VH comprising the amino acid sequence of SEQ ID NO: 56 and a VL comprising the amino acid sequence of SEQ ID NO: 57.
  4. 4 . The iNK cell of any one of claims 1-3 , wherein the CAR comprises a VH comprising the amino acid sequence of SEQ ID NO: 47 and a VL comprising the amino acid sequence of SEQ ID NO: 48.
  5. 5 . The iNKT cell of any one of claims 1-4 , wherein the CAR comprises: (a) a scFv comprising the amino acid sequence of SEQ ID NO: 9; (b) a scFv comprising the amino acid sequence of SEQ ID NO: 16; (c) a scFv comprising the amino acid sequence of SEQ ID NO: 46; (d) a scFv comprising the amino acid sequence of SEQ ID NO: 49; or (e) a scFv comprising the amino acid sequence of SEQ ID NO: 58.
  6. 6 . The iNKT cell of any one of claims 1 - 6 , wherein the CAR comprises a scFv comprising the amino acid sequence of SEQ ID NO: 49.
  7. 7 . The iNKT cell of any one of claims 1-6 , wherein the CAR further comprises a hinge region.
  8. 8 . The iNKT cell of claim 7 , wherein the hinge region comprises an amino acid sequence at least 80% identical to SEQ ID NO: 30.
  9. 9 . The iNKT cell of any one of claims 1-8 , wherein the CAR further comprises a transmembrane domain.
  10. 10 . The iNKT cell of claim 9 , wherein the transmembrane domain comprises an amino acid sequence at least 80% identical to SEQ ID NO: 31.
  11. 11 . The iNKT cell of any one of claims 1-7 , wherein the CAR comprises a hinge/transmembrane domain comprising the amino acid sequence of any one of SEQ ID NOs: 83, 85, or 87.
  12. 12 . The iNKT cell of any of claims 1-11 , wherein the CAR further comprises one or more cytoplasmic domain.
  13. 13 . The iNKT cell of claim 12 wherein the one or more cytoplasmic domain comprises the amino acid sequence of any one of SEQ ID NO: 32, 33, 39, 90, 92, 94, 96, 98, 100, 102, or 105.
  14. 14 . The iNKT cell of claim 12 or 13 , wherein the cytoplasmic domain comprises an amino acid sequence at least 80% identical to SEQ ID NO: 32.
  15. 15 . The iNKT cell of any one of claims 1-14 , wherein the CAR further comprises a intracellular co-stimulatory domain.
  16. 16 . The iNKT cell of claim 15 , wherein the co-stimulatory domain comprises an amino acid sequence at least 80% identical to SEQ ID NO: 33.
  17. 17 . The iNKT cell of any one of claims 1-16 , wherein the CAR comprises the amino acid sequence of any one of SEQ ID NOs: 17, 18, 59, 60, or 62.
  18. 18 . The iNKT cell of any one of claims 1-17 , wherein the CAR comprises the amino acid sequence of SEQ ID NO: 60.
  19. 19 . The iNKT cell of any one of claims 1-18 , wherein the iNKT cell is engineered to express one or more immunoregulatory gene products.
  20. 20 . The iNKT cell of claim 19 , wherein the one or more immunoregulatory gene products comprises IL-15, IL-12, CD40L, or 4-1BB, IL-18, or IL-21.

Description

RELATED APPLICATIONS This application claims the benefit under 35 U.S.C. 119(e) of the filing date of U.S. provisional Application Ser. No. 63/413,236, filed Oct. 4, 2022, the entire contents of which are incorporated by reference herein. REFERENCE TO AN ELECTRONIC SEQUENCE LISTING The contents of the electronic sequence listing (A132770004WO00-SEQ-LGE.xml; Size: 179,363 bytes; and Date of Creation: Oct. 4, 2023) is herein incorporated by reference in its entirety. BACKGROUND Fibroblast activation protein (FAP) is a cell surface protein high expressed on stroma cells (e.g., cancer-associated fibroblasts (CAFs) in the tumor microenvironment (TME). FAP can modulate the TME by remodeling the extracellular matrix (ECM), and its overexpression on CAFs is associated with poor prognosis in various cancers. SUMMARY The present disclosure, at least in part, is based on the discovery that genetically modified cells (e.g., iNKT cells) expressing chimeric antigen receptors targeting fibroblast activation protein (FAP) are capable of killing FAP expressing cells (e.g., cancer-associated fibroblasts (CAFs)) in the tumor microenvironment (TME). In some embodiments, the anti-FAP CAR iNKT cell is engineered to express an armoring molecule (e.g., soluble IL-15). In some embodiments, the anti-FAP CAR iNKT cells kill FAP expressing cells (e.g., FAP expressing tumor cells and/or FAP expressing cancer-associated fibroblasts (CAFs)). In some embodiments, the anti-FAP CAR iNKT cells reduce the immunosuppression in the TME, thereby increasing the efficacy of other cancer therapy (e.g., CAR T cell targeting tumor antigen) In some embodiments, anti-FAP CAR iNKT cells described by the disclosure demonstrate improved properties relative to existing FAP-CAR cell therapy in the art, including but not limited to killing of FAP-expressing cells in vitro and in vivo; and enhanced persistence in a subject receiving the therapy. In some aspects, the present disclosure provides an invariant natural killer T (iNKT) cell comprising a chimeric antigen receptor (CAR) that specifically binds fibroblast activation protein (FAP). In some embodiments, the chimeric antigen receptor of the iNKT cells comprises a CDRH1 having the amino acid sequence of SEQ ID NO: 1, a CDRH2 having the amino acid sequence of SEQ ID NO: 2, a CDRH3 having the amino acid sequence of SEQ ID NO: 3, a CDRL1 having the amino acid sequence of SEQ ID NO: 4, a CDRL2 having the amino acid sequence of SEQ ID NO: 5, and a CDRL3 having the amino acid sequence of SEQ ID NO: 6. In some embodiments, the chimeric antigen receptor of the iNKT cells comprises a CDRH1 having the amino acid sequence of SEQ ID NO: 10, a CDRH2 having the amino acid sequence of SEQ ID NO: 11, a CDRH3 having the amino acid sequence of SEQ ID NO: 12, a CDRL1 having the amino acid sequence of SEQ ID NO: 4, a CDRL2 having the amino acid sequence of SEQ ID NO: 5, and a CDRL3 having the amino acid sequence of SEQ ID NO: 13. In some embodiments, the chimeric antigen receptor of the iNKT cells comprises a CDRH1 having the amino acid sequence of SEQ ID NO: 40, a CDRH2 having the amino acid sequence of SEQ ID NO: 41, a CDRH3 having the amino acid sequence of SEQ ID NO: 42, a CDRL1 having the amino acid sequence of SEQ ID NO: 4, a CDRL2 having the amino acid sequence of SEQ ID NO: 5, and a CDRL3 having the amino acid sequence of SEQ ID NO: 43. In some embodiments, the chimeric antigen receptor of the iNKT cells comprises a CDRH1 having the amino acid sequence of SEQ ID NO: 10, a CDRH2 having the amino acid sequence of SEQ ID NO: 11, a CDRH3 having the amino acid sequence of SEQ ID NO: 42, a CDRL1 having the amino acid sequence of SEQ ID NO: 4, a CDRL2 having the amino acid sequence of SEQ ID NO: 5, and a CDRL3 having the amino acid sequence of SEQ ID NO: 43. In some embodiments, the chimeric antigen receptor of the iNKT cells comprises a CDRH1 having the amino acid sequence of SEQ ID NO: 10, a CDRH2 having the amino acid sequence of SEQ ID NO: 11, a CDRH3 having the amino acid sequence of SEQ ID NO: 54, a CDRL1 having the amino acid sequence of SEQ ID NO: 4, a CDRL2 having the amino acid sequence of SEQ ID NO: 5, and a CDRL3 having the amino acid sequence of SEQ ID NO: 55. In some embodiments, the chimeric antigen receptor of the iNKT cells comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 7 and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 8. In some embodiments, the chimeric antigen receptor of the iNKT cells comprises a VH comprising the amino acid sequence of SEQ ID NO: 14 and a VL comprising the amino acid sequence of SEQ ID NO: 15. In some embodiments, the chimeric antigen receptor of the iNKT cells comprises a VH comprising the amino acid sequence of SEQ ID NO: 44 and a VL comprising the amino acid sequence of SEQ ID NO: 45. In some embodiments, the chimeric antigen receptor of the iNKT cells comprises a VH comprising the amino acid seq