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CN-122011203-A - Chimeric antigen receptor with chimeric intracellular signaling domain based on cd3ζ and cd3δ and uses thereof

CN122011203ACN 122011203 ACN122011203 ACN 122011203ACN-122011203-A

Abstract

Chimeric antigen receptors having chimeric intracellular signaling domains based on CD3ζ and CD3δ and uses thereof are disclosed. In particular, the invention provides a Chimeric Antigen Receptor (CAR) comprising, from amino terminus to carboxy terminus, a signal peptide region, an antigen binding domain, a hinge region, a transmembrane domain, a co-stimulatory domain, a signaling domain, connected in sequence, wherein the signaling domain comprises ITAM1 and CD3 delta of CD3 zeta, connected in sequence. The chimeric antigen receptor of the invention can obviously strengthen the killing function and the anti-tumor effect of the CAR-T cells.

Inventors

  • WANG WEN
  • CHU QINGSONG
  • CHEN TING
  • YANG CHUNHUI

Assignees

  • 天宜康医药(上海)有限公司

Dates

Publication Date
20260512
Application Date
20250918

Claims (10)

  1. 1. A Chimeric Antigen Receptor (CAR), comprising, from amino terminus to carboxy terminus, a signal peptide region, an antigen binding domain, a hinge region, a transmembrane domain, a costimulatory domain, a signal transduction domain, all of which are sequentially linked; wherein the signal transduction domain comprises ITAM1 and cd3δ of cd3ζ connected in sequence.
  2. 2. The chimeric antigen receptor according to claim 1, wherein the amino acid sequence of ITAM1 of CD3 zeta is shown in SEQ ID NO. 24 and the amino acid sequence of CD3 delta is shown in SEQ ID NO. 26.
  3. 3. The chimeric antigen receptor of claim 1, wherein the chimeric antigen receptor is a single-target chimeric antigen receptor or a dual-target chimeric antigen receptor.
  4. 4. The chimeric antigen receptor of claim 3, wherein when the chimeric antigen receptor is a dual-target chimeric antigen receptor, the antigen binding domain comprises a first antigen binding domain and a second antigen binding domain, wherein the first antigen binding domain and the second antigen binding domain are sequentially linked by a linker; And, the first antigen binding domain binds ：CD19、CD7、CD2、CD3、CD4、CD5、CD8、CD20、CD22、CD25、CD28、CD30、CD33、CD38、CD40、CD44V6、CD47、CD52、CD56、CD57、CD58、CD79b、CD80、CD86、CD81、CD123、CD133、CD151、CD171、CD276、CLL1、B7H4、BCMA、VEGFR-2、EGFR、GPC3、PMSA、Her2、Mesothelin、CS1、MUC16、GD2、CEA、CD138、EGFR、EGFRVIII、LewisY、DLL3、GPRC5D、MG7、IL13Rα2、 or a combination thereof to an antigen selected from the group consisting of; the second antigen binding domain binds ：CD19、CD7、CD2、CD3、CD4、CD5、CD8、CD20、CD22、CD25、CD28、CD30、CD33、CD38、CD40、CD44V6、CD47、CD52、CD56、CD57、CD58、CD79b、CD80、CD86、CD81、CD123、CD133、CD151、CD171、CD276、CLL1、B7H4、BCMA、VEGFR-2、EGFR、GPC3、PMSA、Her2、Mesothelin、CS1、MUC16、GD2、CEA、CD138、EGFR、EGFRVIII、LewisY、DLL3、GPRC5D、MG7、IL13Rα2、 or a combination thereof to an antigen selected from the group consisting of.
  5. 5. A conjugate, characterized in that, the conjugate comprises: (a) The chimeric antigen receptor of claim 1, and (B) A coupling moiety coupled to the chimeric antigen receptor, the coupling moiety being a detectable label.
  6. 6. A nucleic acid molecule encoding the chimeric antigen receptor of claim 1.
  7. 7. A vector comprising the nucleic acid molecule of claim 6.
  8. 8. A genetically engineered host cell comprising the vector of claim 7, or having integrated into its genome the nucleic acid molecule of claim 6, or expressing the chimeric antigen receptor of claim 1.
  9. 9. Use of a chimeric antigen receptor according to claim 1, a nucleic acid molecule according to claim 6, a vector according to claim 7, a host cell according to claim 8 for the preparation of a medicament for the treatment of an immune-related disorder, which is an autoimmune disorder or a tumor.
  10. 10. A pharmaceutical composition comprising the expression vector of claim 7 or the host cell of claim 8.

Description

Chimeric antigen receptor with chimeric intracellular signaling domain based on cd3ζ and cd3δ and uses thereof Technical Field The present invention is in the field of immunotherapy, in particular to chimeric antigen receptors with chimeric intracellular signaling domains based on cd3ζ and cd3δ and uses thereof. Background Chimeric antigen receptor T cell (CAR-T) cell therapy is an emerging immune cell therapy, the safety and efficacy of which has been widely demonstrated in a variety of blood tumor types and in part in solid tumors. In a partial hematological tumor patient, the patient can remain cancer-free for years or even heal after CAR-T cell therapy. Early CAR-T cell therapies have a number of limitations, such as cytokine storm (CRS) due to CAR-T cell overactivation, neurotoxicity due to CAR molecule off-target (on target off tumor), and immunosuppression and premature depletion of T cell function in solid tumors. In recent years, with advances in technology, the safety risk of CAR-T cells has been greatly improved, but CAR-T treatment of solid tumors still faces a great challenge. Optimization of chimeric antigen receptor CAR structure is of great benefit for improving CAR-T therapeutic effect, reducing side effects. The CAR molecular structure skeleton mainly consists of antigen binding domain (anti binder), hinge region (hinge), transmembrane domain (TM), intracellular co-stimulatory domain (ICS), and signal transduction domain (CD 3 z) functional elements. It is generally believed that the affinity of the antigen binding domain correlates with the activation efficiency of the CAR molecule, and that the transmembrane domain may affect the sensitivity of antigen recognition, while the intracellular co-stimulatory domain correlates with a variety of cellular functions, such as cytokine secretion, cell proliferation efficiency, CD4/CD8 ratio, in vivo persistence, and the like. Currently, fewer attempts are made to engineer optimization strategies for the signaling domain (CD 3 z) in the CAR molecular scaffold. Thus, it would be of great significance in the art to develop improved methods of signaling domains for CARs, thereby obtaining novel signaling domains to achieve an improvement in the therapeutic efficacy of CAR-T. Disclosure of Invention The invention provides a chimeric intracellular signaling domain based on CD3δ and CD3ζ and uses thereof in CAR-T cells. In a first aspect of the invention, there is provided a Chimeric Antigen Receptor (CAR) comprising, from amino-terminus to carboxy-terminus, a signal peptide region, an antigen binding domain, a hinge region, a transmembrane domain, a co-stimulatory domain, a signal transduction domain, connected in sequence; wherein the signal transduction domain comprises ITAM1 and cd3δ of cd3ζ connected in sequence. In another preferred embodiment, the amino acid sequence of ITAM1 of CD3 ζ is shown in SEQ ID NO. 24. In another preferred embodiment, the amino acid sequence of CD3 delta is shown in SEQ ID NO. 26. In another preferred embodiment, the chimeric antigen receptor is a single-target chimeric antigen receptor or a dual-target chimeric antigen receptor. In another preferred embodiment, the signal peptide is a CD8 leader peptide. In another preferred embodiment, the amino acid sequence of the CD8 leader is shown in SEQ ID NO. 2. In another preferred embodiment, when the chimeric antigen receptor is a single-target chimeric antigen receptor, the antigen binding domain binds to an antigen selected from the group consisting of ：CD19、CD7、CD2、CD3、CD4、CD5、CD8、CD20、CD22、CD25、CD28、CD30、CD33、CD38、CD40、CD44V6、CD47、CD52、CD56、CD57、CD58、CD79b、CD80、CD86、CD81、CD123、CD133、CD151、CD171、CD276、CLL1、B7H4、BCMA、VEGFR-2、EGFR、GPC3、PMSA、Her2、Mesothelin、CS1、MUC16、GD2、CEA、CD138、EGFR、EGFRVIII、LewisY、DLL3、GPRC5D、MG7、IL13Rα2、 or a combination thereof. In another preferred embodiment, when the chimeric antigen receptor is a dual-target chimeric antigen receptor, the antigen binding domain comprises a first antigen binding domain and a second antigen binding domain. In another preferred embodiment, the first antigen binding domain and the second antigen binding domain are linked in sequence by a linker. In another preferred embodiment, the first antigen binding domain binds ：CD19、CD7、CD2、CD3、CD4、CD5、CD8、CD20、CD22、CD25、CD28、CD30、CD33、CD38、CD40、CD44V6、CD47、CD52、CD56、CD57、CD58、CD79b、CD80、CD86、CD81、CD123、CD133、CD151、CD171、CD276、CLL1、B7H4、BCMA、VEGFR-2、EGFR、GPC3、PMSA、Her2、Mesothelin、CS1、MUC16、GD2、CEA、CD138、EGFR、EGFRVIII、LewisY、DLL3、GPRC5D、MG7、IL13Rα2、 or a combination thereof to an antigen selected from the group consisting of. In another preferred embodiment, the second antigen binding domain binds ：CD19、CD7、CD2、CD3、CD4、CD5、CD8、CD20、CD22、CD25、CD28、CD30、CD33、CD38、CD40、CD44V6、CD47、CD52、CD56、CD57、CD58、CD79b、CD80、CD86、CD81、CD123、CD133、CD151、CD171、CD276、CLL1、B7H4、BCMA、VEGFR-2、EGFR、GPC3、PMSA、Her2、Mesothelin、CS1、MUC16、GD2、CEA、CD138、EGFR、EGFRVIII、LewisY、DLL3、GPRC5D、MG7、IL13Rα2、 or a combinati