Chimeric antigen receptor and CAR-T cells that bind a herpes virus antigen

Patent metadata

Field	Value
Publication number	US-12054529-B2
Application number	US-201917048928-A
Country	US
Kind code	B2
Filing date	Apr 17, 2019
Priority date	Apr 20, 2018
Publication date	Aug 6, 2024
Grant date	Aug 6, 2024

Abstract

An isolated chimeric antigen receptor (CAR) polypeptide, wherein the CAR includes an extracellular antigen-binding domain, including an antibody or antibody fragment that binds to a protein encoded by a herpes virus, or to a protein complex including the protein (herpes virus antigen), wherein the herpes virus antigen is present on the surface of a human cell that is latently infected with said herpes virus and supports the lytic phase of viral replication. The invention further relates to a nucleic acid molecule encoding the CAR of the invention, a genetically modified immune cell, preferably a T cell, expressing the CAR of the invention and the use of the cell in the treatment of a medical disorder associated with human herpesvirus, such as herpes virus-associated cancers, chronic active herpes virus infections or primary herpes virus infections. In preferred embodiments the herpes virus is Epstein-Barr virus (EBV) and a preferred herpes virus antigen target of the CAR is the EBV glycoprotein 350/220 (gp350/gp220).

First claim

The invention claimed is:

1. A chimeric antigen receptor polypeptide (CAR), comprising:
i. an extracellular antigen-binding domain, comprising an antibody or antibody fragment that binds to an Epstein-Barr virus (EBV) glycoprotein 350/220 (gp350/gp220),
ii. a transmembrane domain, and
iii. an intracellular signaling domain,
wherein the antigen-binding domain comprises a variable heavy chain (VH), said VH comprising heavy chain complementary determining regions H-CDR1 according to SEQ ID NO: 1 (GLSLTSN), H-CDR2 according to SEQ ID NO: 2 (WSNGG), and H-CDR3 according to SEQ ID NO: 3 (PRYNSGYFFDY), and a variable light chain (VL), said VL comprising light chain complementary determining regions L-CDR1 according to SEQ ID NO: 4 (KASESVSTRMH), L-CDR2 according to SEQ ID NO: 5 (KTSNLAS), and L-CDR3 according to SEQ ID NO: 6 (QQSWNGPLT), or
wherein the antigen-binding domain comprises a variable heavy chain (VH), said VH comprising heavy chain complementary determining regions H-CDR1 according to SEQ ID NO: 7 (GFSLTSY), H-CDR2 according to SEQ ID NO: 8 (WSDGD), and H-CDR3 according to SEQ ID NO: 9 (LQSEDTATYYCARLQVFGYPGIRDYVMDA), and a variable light chain (VL), said VL comprising light chain complementary determining regions L-CDR1 according to SEQ ID NO: 10 (KSSQSLLSSRHQKNFLA), L-CDR2 according to SEQ ID NO: 11 (HASTRQS), and L-CDR3 according to SEQ ID NO: 12 (LQHYTSPYT).

2. The CAR polypeptide according to claim 1 , wherein the EBV glycoprotein gp350/gp220 is present on the surface of EBV-infected cells.

3. The CAR polypeptide according to claim 2 , wherein the EBV-infected cells are selected from the group consisting of EBV-infected cancer cells, EBV-infected B cells and EBV-infected epithelial cells.

4. The CAR polypeptide according to claim 1 :
wherein the CAR comprises a leader polypeptide positioned N-terminally of the VH and VL domains, and/or
wherein the extracellular antigen-binding domain comprises a linker polypeptide positioned between the VH and VL domains, and/or
comprising additionally a spacer polypeptide positioned between the extracellular antigen-binding domain and the transmembrane domain, and/or
wherein the transmembrane domain is a CD28 or a CD8 alpha transmembrane domain; and/or
wherein the intracellular domain comprises a CD28 or a 4-1BB co-stimulatory domain; and/or
wherein the intracellular domain comprises a CD3 zeta chain signaling domain; and/or
wherein the CAR comprises one or more linker polypeptides positioned between the VH and VL domains and the spacer, and/or between the spacer and the transmembrane domain.

5. The CAR polypeptide according to claim 4 , wherein the leader polypeptide is an IgHL leader, wherein the linker is a G4S linker, and wherein the spacer is an IgG1 CH3 or a IgG1 CH2-CH3 spacer.

6. An isolated nucleic acid molecule, comprising a nucleotide sequence which encodes a CAR polypeptide according to claim 1 .

7. A genetically modified immune cell comprising a nucleic acid molecule according to claim 6 and/or expressing a CAR according to claim 1 .

8. The genetically modified immune cell according to claim 7 , wherein the immune cell is selected from the group consisting of a T lymphocyte, an NK cell, a macrophage, a dendritic cell, a cytotoxic T lymphocyte and a T helper cell.

9. A method for the treatment of a medical condition associated with an EBV infection in a subject comprising administering to the subject a genetically modified immune cell according to claim 7 .

10. The method according to claim 9 , wherein the medical condition is an EBV-associated cancer.

11. The method according to claim 10 , wherein the medical condition is selected from the group consisting of a lymphoproliferative disorder (LPD), B-cell lymphoma, Burkitt lymphoma (BL), Hodgkin lymphoma (HL), a diffuse large B cell lymphoma (DLBCL), a post-transplant lymphoproliferative disorder (PTLD), an epithelial carcinoma, a lymphoepithelioma, a carcinoma with lymphoid stroma and a glioma.

12. The method according to claim 9 , wherein the medical condition associated with EBV infection is chronic active EBV infection (CAEBV) or primary EBV infection.

13. The method according to claim 9 for use in the treatment of immune deficient or immune compromised patients after chemotherapy, radiation, immune suppression or transplantation.

Assignees

• Medizinische Hochschule Hannover
• Helmholtz Zentrum Muenchen—Deutsches Forschungszentrum Fuer Gesundheit Und Umwelt Gmbh

Inventors

• Stripecke Renata
• Slabik Constanze
• Zeidler Reinhard
• Hammerschmidt Wolfgang

Classifications

• A61K40/46

  Viral antigens · CPC title

• A61K40/31

  Chimeric antigen receptors [CAR] · CPC title

• A61K40/11

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

• A61K2239/38

  characterised by the dose, timing or administration schedule · CPC title

• A61K2239/31

  characterized by the route of administration · CPC title