阅读说明：本技术 一种嵌合抗原受体及其应用 (Chimeric antigen receptor and application thereof ) 是由 杨选明 傅阳心 李范林 于 2020-01-20 设计创作，主要内容包括：本申请提供一种嵌合抗原受体(CAR),其中所述CAR包含靶向部分、跨膜结构域、共刺激结构域和胞内信号传导结构域,其中所述靶向部分特异性结合T细胞受体(TCR)的可变区Vα、Vβ和/或它们的片段。(The present application provides a Chimeric Antigen Receptor (CAR), wherein the CAR comprises a targeting moiety, a transmembrane domain, a costimulatory domain, and an intracellular signaling domain, wherein the targeting moiety specifically binds to the variable regions va, ν β, and/or fragments thereof of a T Cell Receptor (TCR).)

A Chimeric Antigen Receptor (CAR), wherein the CAR comprises a targeting moiety, a transmembrane domain, a costimulatory domain, and an intracellular signaling domain, wherein the targeting moiety specifically binds to the variable region va, ν β, and/or fragments thereof of a T Cell Receptor (TCR).

The CAR of claim 1, wherein the targeting moiety specifically binds to the variable region V α or a fragment thereof of the TCR.

The CAR of claim 2, wherein the V α or fragment thereof comprises V α 24 or a fragment thereof.

The CAR of any one of claims 2-3, wherein the V α or fragment thereof further comprises J α or fragment thereof.

The CAR of claim 4, wherein the J α or fragment thereof comprises J α 18 or fragment thereof.

The CAR of claim 1, wherein the targeting moiety specifically binds to the variable region V β of the TCR, or a fragment thereof.

The CAR of claim 6, wherein the V β or fragment thereof comprises V β 8 or a fragment thereof.

The CAR of any one of claims 1-7, wherein the fragment of the TCR variable region comprises the CDR3 of the TCR variable region.

The CAR of any one of claims 1-8, wherein the variable regions of the TCR V α, V β and/or fragments thereof comprise the amino acid sequence of SEQ ID NO: 58-61.

The CAR of any one of claims 1-9, wherein the targeting moiety comprises an antibody or fragment thereof.

The CAR of claim 10, wherein the antibody competes for binding to the variable regions va, ν β, and/or fragments thereof of the TCR with a reference antibody, wherein the reference antibody comprises a light chain variable region comprising LCDR1, LCDR2 and LCDR3, the LCDR1 comprising SEQ ID NO: 3. SEQ ID NO: 64 and SEQ ID NO: 21; the LCDR2 comprises SEQ ID NO: 4. SEQ ID NO: 65 and SEQ ID NO: 22; the LCDR3 comprises SEQ ID NO: 5. SEQ ID NO: 66 and SEQ ID NO: 23; and the number of the first and second electrodes,

the heavy chain variable region of the reference antibody comprises HCDR1, HCDR2, and HCDR3, the HCDR1 comprises SEQ ID NO: 11. SEQ ID NO: 67 and SEQ ID NO: 29; the HCDR2 comprises SEQ ID NO: 12. SEQ ID NO: 68 and SEQ ID NO: 30; the HCDR3 comprises SEQ ID NO: 13. SEQ ID NO: 69 and SEQ ID NO: 31, or a pharmaceutically acceptable salt thereof.

The CAR of claim 11, wherein the light chain variable region of the reference antibody comprises the amino acid sequence of SEQ ID NO: 9. SEQ ID NO: 70 and SEQ ID NO: 27, or a pharmaceutically acceptable salt thereof; and the heavy chain variable region of the reference antibody comprises SEQ ID NO: 17. SEQ ID NO: 71 and SEQ ID NO: 35, or a pharmaceutically acceptable salt thereof.

The CAR of any one of claims 11-12, wherein the reference antibody comprises a va 24 ja 18 antibody.

The CAR of any one of claims 10-13, wherein the antibody comprises heavy chain complementarity determining region 1(HCDR1), the HCDR1 comprising SEQ ID NO: 11. SEQ ID NO: 67 and SEQ ID NO: 29, or a pharmaceutically acceptable salt thereof.

The CAR of any one of claims 10-14, wherein the antibody further comprises heavy chain complementarity determining region 2(HCDR2), the HCDR2 comprising SEQ ID NO: 12. SEQ ID NO: 68 and SEQ ID NO: 30, or a pharmaceutically acceptable salt thereof.

The CAR of any one of claims 10-15, wherein the antibody further comprises heavy chain complementarity determining region 3(HCDR3), the HCDR3 comprising SEQ ID NO: 13. SEQ ID NO: 69 and SEQ ID NO: 31, or a pharmaceutically acceptable salt thereof.

The CAR of any one of claims 10-16, wherein the antibody comprises light chain complementarity determining region 1(LCDR1), the LCDR1 comprising SEQ ID NO: 3. SEQ ID NO: 64 and SEQ ID NO: 21, or a pharmaceutically acceptable salt thereof.

The CAR of any one of claims 10-17, wherein the antibody further comprises light chain complementarity determining region 2(LCDR2), the LCDR2 comprising SEQ ID NO: 4. SEQ ID NO: 65 and SEQ ID NO: 22, or a pharmaceutically acceptable salt thereof.

The CAR of any one of claims 10-18, wherein the antibody further comprises light chain complementarity determining region 3(LCDR3), the LCDR3 comprising SEQ ID NO: 5. SEQ ID NO: 66 and SEQ ID NO: 23, or a pharmaceutically acceptable salt thereof.

The CAR of any one of claims 10-19, wherein the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 17. SEQ ID NO: 71 and SEQ ID NO: 35, or a pharmaceutically acceptable salt thereof.

The CAR of any one of claims 10-20, wherein the antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 9. SEQ ID NO: 70 and SEQ ID NO: 27, or a pharmaceutically acceptable salt thereof.

The CAR of any one of claims 10-21, wherein the antibody is a single chain antibody.

The CAR of any one of claims 10-22, wherein the antibody comprises the amino acid sequence of SEQ ID NO: 1. SEQ ID NO: 62 and SEQ ID NO: 19, or a pharmaceutically acceptable salt thereof.

The CAR of any one of claims 1-23, wherein the transmembrane domain comprises a polypeptide derived from a protein selected from the group consisting of: CD8, CD28, CD137, and/or CD 3.

The CAR of any one of claims 1-24, wherein the transmembrane domain comprises the amino acid sequence of SEQ ID NO: 40, or a pharmaceutically acceptable salt thereof.

The CAR of any one of claims 1-25, wherein the co-stimulatory domain comprises a polypeptide derived from a protein selected from the group consisting of: ICOS, CD28, CD27, HVEM, LIGHT, CD40L, 4-1BB, OX40, DR3, GITR, CD30, TIM1, SLAM, CD2, and/or CD 226.

The CAR of any one of claims 1-26, wherein the co-stimulatory domain comprises the amino acid sequence of SEQ ID NO: 42.

The CAR of any one of claims 1-27, wherein the intracellular signaling domain comprises a signaling domain from CD3 ζ.

The CAR of any one of claims 1-28, wherein the intracellular signaling domain comprises SEQ ID NO: 44, or a pharmaceutically acceptable salt thereof.

The CAR of any one of claims 1-29, wherein the CAR further comprises a hinge region that connects the antibody and the transmembrane domain.

The CAR of claim 30, wherein the hinge region comprises a polypeptide derived from a protein selected from the group consisting of: CD8a and/or IgG.

The CAR of any one of claims 30-31, wherein the hinge region comprises the amino acid sequence of SEQ ID NO: 38, or a pharmaceutically acceptable salt thereof.

The CAR of any one of claims 1-32, comprising the amino acid sequence of SEQ ID NO: 46. SEQ ID NO: 72 and SEQ ID NO: 50.

An isolated nucleic acid molecule encoding the CAR of any one of claims 1-33.

An isolated nucleic acid molecule encoding a CAR comprising SEQ ID NO: 47. SEQ ID NO: 73 or SEQ ID NO: 51.

A vector comprising the nucleic acid molecule of any one of claims 34-35.

The vector of claim 36, comprising an mRNA vector, a plasmid vector, a retroviral vector, and/or a lentiviral vector.

An immune effector cell comprising and/or capable of expressing a CAR of any one of claims 1-33, a nucleic acid molecule of any one of claims 34-35, or a vector of any one of claims 36-37.

The cell of claim 38, wherein the immune effector cell is selected from a T lymphocyte and a Natural Killer (NK) cell.

The cell of claim 39, wherein the T lymphocyte and/or natural killer cell is of natural origin, or is induced by a stem cell.

A method of making an immune effector cell, comprising introducing into an immune effector cell the vector of any one of claims 36-37.

A composition comprising the immune effector cell of any one of claims 38-40.

Use of the CAR of any one of claims 1-33, the nucleic acid molecule of any one of claims 34-35, the vector of any one of claims 36-37, or the immune effector cell of any one of claims 38-40 for the manufacture of a medicament, wherein the medicament is for treating a T-cell lymphoma or leukemia.

The use of claim 43, wherein the T-cell lymphoma or leukemia comprises one or more selected from the group consisting of: peripheral T cell lymphoma, non-specific peripheral T cell lymphoma, angioimmunoblastic lymphoma, extranodal NKT cell lymphoma, anaplastic T cell lymphoma, ALK-positive anaplastic cell lymphoma, ALK-negative anaplastic cell lymphoma, T lymphoblastic lymphoma, cytotoxic T cell lymphoma, cutaneous T cell lymphoma, adult T cell leukemia/lymphoma (ATLL), T prolymphocytic leukemia (T-PLL), T large granular lymphocyte (T-LGL) leukemia, hepatosplenic T cell lymphoma (HSTL), Sezary Syndrome (SS), subcutaneous tonsillar T cell lymphoma, and undifferentiated T cell lymphoma.