Granted Patent B2

US 12,454,564 · App. 17/821,664 · Granted Oct 28, 2025

Inventors: Xinxin Wang (Shanghai, CN); Tao Jin (Shanghai, CN); Chunhui Yang (Shanghai, CN); Zhongdong Shi (Shanghai, CN); Liping Liu (Shanghai, CN); Jing Sun (Shanghai, CN); Shuyi Qiu (Shanghai, CN); Wei Cao (Shanghai, CN)

Assignees: Gracell Biotechnologies (Shanghai) Co., Ltd.; Suzhou Gracell Biotechnologies Co., Ltd.

C07K14/7051A61K40/11A61K40/31A61K40/4202A61K40/4204A61K40/421A61K40/4211A61K40/4215A61P35/00C07K14/70507C07K14/70596C12N5/0636A61K38/00A61K40/50A61K2239/23A61K2239/29A61K2239/31A61K2239/38A61K2239/48

Abstract

Claims (34)

1. A method for treating cancer in a human subject while reducing host-versus-graft (HvG) rejection by an innate T cell or NK cell in the human subject, the method comprising:

administering an engineered cytotoxic T cell to the human subject, wherein the engineered cytotoxic T cell comprises one or more chimeric antigen receptors (CARs) comprising:

a first antigen binding moiety exhibiting specific binding to CD7, wherein the first antigen binding moiety comprises a first scFv comprising a heavy chain variable domain (VH1) comprising the polypeptide sequence of SEQ ID NO: 91 and a light chain variable domain (VL1) comprising the polypeptide sequence of SEQ ID NO: 90; and

a second antigen binding moiety exhibiting specific binding to CD19, wherein the second antigen binding domain comprises a second scFv comprising a heavy chain variable domain (VH2) comprising the polypeptide sequence of SEQ ID NO: 88 and a light chain variable domain (VL2) comprising the polypeptide sequence of SEQ ID NO: 87,

wherein each CAR of the one or more CARs further comprises:

a hinge polypeptide, which is C-terminal to the first antigen binding moiety and the second antigen-binding moiety;

a transmembrane polypeptide, which is C-terminal to the hinge polypeptide; and

an intracellular signaling polypeptide comprising a signaling domain of CD3-zeta, which is C-terminal to the transmembrane polypeptide.

2. The method of claim 1 , wherein the first antigen binding moiety and the second antigen binding moiety are arranged, from the amino terminus to the carboxyl terminus, according to one of the following formulas (I-a)-(I-f):

the VL2- the VL1- the VH1- the VH2 (I-a);

the VH2- the VH1- the VL1- the VL2 (I-b);

the VL1- the VL2- the VH2- the VH1 (I-c);

the VH1- the VH2- the VL2- the VL1 (I-d);

the VL2- the VH2- the VH1- the VH1 (I-e); and

the VL1- the VH1- the VH2- the VL2 (I-f).

3. The method of claim 1 , wherein the first antigen binding moiety and the second antigen binding moiety are arranged, from the amino terminus to the carboxyl terminus, according to one of the following formulas (II-a)-(II-j):

the VL2- the VH1- the VL1- the VH2 (II-a);

the VH2- the VL1- the VH1- the VL2 (II-b);

the VL1- the VH2- the VL2- the VH1 (II-c);

the VH1- the VL2- the VH2- the VL1 (II-d);

the VL2- the VH2- the VL1- the VH1 (II-e);

the VL1- the VH1- the VL2- the VH2 (II-f);

the VH2- the VL2- the VL1- the VH1 (II-g);

the VH2- the VL2- the VH1- the VL1 (II-h);

the VH1- the VL1- the VL2- the VH2 (II-i); and

the VH1- the VL1- the VH2- the VL2 (II-j).

4. The method of claim 1 , wherein, in the engineered cytotoxic T cell, an endogenous gene encoding CD7 is knocked out or silenced, to induce fratricide resistance.

5. The method of claim 1 , wherein a subunit of an endogenous T cell receptor (TCR) of the engineered cytotoxic T cell is knocked out or silenced, to reduce graft-versus-host disease (GVHD) effect of the engineered cytotoxic T cell in the human subject.

6. The method of claim 5 , wherein the subunit is TCR alpha.

7. The method of claim 1 , wherein the engineered cytotoxic T cell further comprises a heterologous polypeptide comprising the polypeptide sequence of SEQ ID NO: 11.

8. The method of claim 1 , wherein the engineered cytotoxic T cell further comprises a heterologous polypeptide comprising the polypeptide sequence of SEQ ID NO: 15.

9. The method of claim 1 , wherein the engineered cytotoxic T cell further comprises a heterologous polypeptide comprising the polypeptide sequence of SEQ ID NO: 17.

10. The method of claim 1 , wherein the first scFV comprises the polypeptide sequence of SEQ ID NO: 75.

11. The method of claim 1 , wherein the intracellular signaling polypeptide further comprises a costimulatory signaling domain, wherein the costimulatory signaling domain comprises a signaling domain of 4-1BB, a signaling domain of CD28, or the signaling domains of both 4-1BB and CD28.

Assignments (3)

Priority Claims (5)

Continuity (3)

References Cited (239)