Humanized BCMA antibody and BCMA-CAR-T cells

Patent metadata

Field	Value
Publication number	US-11472884-B2
Application number	US-202217700160-A
Country	US
Kind code	B2
Filing date	Mar 21, 2022
Priority date	Jan 16, 2019
Publication date	Oct 18, 2022
Grant date	Oct 18, 2022

Abstract

The present invention is directed to a humanized BCMA single-chain variable fragment (scFv), comprising V H having the amino acid sequence of SEQ ID NO: 3 and V L having the amino acid sequence of SEQ ID NO: 5. The present invention is also directed to a BCMA chimeric antigen receptor fusion protein comprising from N-terminus to C-terminus: (i) a single-chain variable fragment (scFv) of the present invention, (ii) a transmembrane domain, (iii) at least one co-stimulatory domains, and (iv) an activating domain. This humanized BCMA-CAR-T cells have specific killing activity against BCMA-positive tumor cells.

First claim

What is claimed is:

1. A method of decreasing growth of a BCMA-positive tumor, the method comprising contacting the tumor with engineered immune cells expressing an anti-BCMA chimeric antigen receptor (CAR) having an anti-BCMA single-chain variable fragment (scFv) comprising:
a. a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 3, and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 5;
b. a transmembrane domain selected from the group consisting of a T cell receptor α chain, a T cell receptor β chain, a CD3 zeta chain, a CD28, a CD3ε, a CD45, a CD4, a CD5, a CD8, a CD9, a CD16, a CD22, a CD33, a CD37, a CD64, a CD80, a CD86, a CD134, a CD137, an ICOS, a CD154, and a GITR;
c. a co-stimulatory domain selected from the group consisting of a CD28, a 4-1BB, a GITR, an ICOS-1, a CD27, an OX-40, and a DAP10; and
d. a CD3zeta activating domain.

2. The method of claim 1 , wherein the engineered immune cells are selected from T-cells, NK cells, macrophages, and hematopoietic cells.

3. The method of claim 1 , wherein the transmembrane domain is a CD28 transmembrane domain, and the costimulatory domain is a 4-1BB costimulatory domain.

4. The method of claim 3 , wherein the CD28 transmembrane domain comprises SEQ ID NO: 12.

5. The method of claim 3 , wherein the CD28 transmembrane domain is encoded by SEQ ID NO: 11.

6. The method of claim 1 , wherein the CD3zeta activating domain comprises SEQ ID NO: 16.

7. The method of claim 6 , wherein the CD3zeta activating domain is encoded by SEQ ID NO: 15.

8. The method of claim 1 , wherein the CAR further comprises a hinge domain.

9. The method of claim 8 , wherein the hinge domain is a CD8 hinge domain.

10. The method of claim 9 , wherein the CD8 hinge domain comprises SEQ ID NO: 10.

11. The method of claim 1 , wherein the CAR further comprises a leader sequence.

12. The method of claim 11 , wherein the leader sequence is a CD8 leader sequence.

13. The method of claim 12 , wherein the CD8 leader sequence comprises SEQ ID NO: 8.

Assignees

• Caribou Biosciences Inc

Inventors

• Wu Lijun
• Golubovskaya Vita

Classifications

• A61P35/00 Primary

  Antineoplastic agents · CPC title

• C07K2317/622

  Single chain antibody (scFv) · CPC title

• A61K2035/124

  the cells being hematopoietic, bone marrow derived or blood cells · CPC title

• C12N2510/00

  Genetically modified cells · CPC title

• C07K14/70517

  CD8 · CPC title