Bispecific antibodies targeting cd22 and cd19 and chimeric antigen receptors and uses thereof
By designing bispecific antibodies and chimeric antigen receptors targeting CD22 and CD19, the problem of relapse caused by antigen mutations in CD19-CAR-T cell therapy was solved, enhancing the killing power against B-cell tumors, reducing the relapse rate, and achieving highly efficient anti-tumor treatment.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- CHONGQING PRECISION BIOTECH CO LTD
- Filing Date
- 2021-12-06
- Publication Date
- 2026-06-26
AI Technical Summary
Current CD19-CAR-T cell therapy for B-ALL can lead to relapse due to antigen mutations or loss, and cannot effectively target B-cell tumors. Furthermore, the low expression level of CD22 makes it difficult to improve treatment efficacy.
We designed bispecific antibodies and chimeric antigen receptors targeting CD22 and CD19, containing specific amino acid sequence combinations and linker structures, to construct dual-target CAR-T cells and enhance their ability to kill B-cell tumors.
It enhances the killing power against B-cell tumors, reduces immune escape, lowers the recurrence rate after treatment, is suitable for industrial production, and has shown significant anti-tumor effects in trials.
Smart Images
Figure CN116217732B_ABST
Abstract
Claims
1. A chimeric antigen receptor targeting CD22 and CD19, characterized in that, Includes one of the following structures: (1) The chimeric antigen receptor is sequentially connected to an extracellular structural domain, a hinge region, a transmembrane region, and an intracellular signaling domain, and its structure is one of the following: (a) CD22VL-linker4-CD22VH-Linker1-CD19VL-linker4-CD19VH-8h-8TM-BBZ; (b) CD19VL-linker4-CD19VH-Linker1-CD22VL-linker4-CD22VH-8h-8TM-BBZ; (c) CD22VL-linker4-CD22VH-Linker2-CD19VL-linker4-CD19VH-8h-8TM-BBZ; (d) CD22VL-linker3-CD19VL-linker4-CD19VH-linker3-CD22VH-8h-8TM-BBZ; (e) CD22VH-linker3-CD19VL-linker4-CD19VH-linker3-CD22VL-8h-8TM-BBZ; The amino acid sequence of CD22VL is shown in SEQ ID NO:1; the amino acid sequence of CD22VH is shown in SEQ ID NO:2; the amino acid sequence of CD19VL is shown in SEQ ID NO:3; and the amino acid sequence of CD19VH is shown in SEQ ID NO:
4. The amino acid sequence of linker4 is shown in SEQ ID NO:5; the amino acid sequence of linker1 is shown in SEQ ID NO:6; the amino acid sequence of linker2 is shown in SEQ ID NO:7; and the amino acid sequence of linker3 is shown in SEQ ID NO:
8. The amino acid sequence of the 8h is shown in SEQ ID NO:15; the amino acid sequence of the 8TM is shown in SEQ ID NO:17; in the BBZ, the amino acid sequence of BB is shown in SEQ ID NO:20, and the amino acid sequence of Z is shown in SEQ ID NO:
19. or (2) CD19VL-linker4-CD19VH-8h_1-8TM-BBZ-2A-CD22VL-linker4-CD22VH-8h_2-28TM-28Z; In the structure (2), the amino acid sequence of CD19VL is shown in SEQ ID NO:3; the amino acid sequence of CD19VH is shown in SEQ ID NO:4; the amino acid sequence of CD22VL is shown in SEQ ID NO:1; the amino acid sequence of CD22VH is shown in SEQ ID NO:2; and the amino acid sequence of linker4 is shown in SEQ ID NO:
5. The amino acid sequence of 8h_1 is shown in SEQ ID NO:15; the amino acid sequence of 8TM is shown in SEQ ID NO:17; in BBZ, the amino acid sequence of BB is shown in SEQ ID NO:20, and the amino acid sequence of Z is shown in SEQ ID NO:
19. The amino acid sequence of 2A is selected from the sequence shown in SEQ ID NO:14, SEQ ID NO:60 or SEQ ID NO:61; The amino acid sequence of 8h_2 is shown in SEQ ID NO:16; the amino acid sequence of 28TM is shown in SEQ ID NO:18; in 28Z, the amino acid sequence of 28 is shown in SEQ ID NO:22, and the amino acid sequence of Z is shown in SEQ ID NO:
21.
2. A nucleic acid molecule, characterized in that, The chimeric antigen receptor of claim 1 has a nucleic acid sequence as shown in SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:85 or SEQ ID NO:
86.
3. The nucleic acid molecule according to claim 2, characterized in that, The nucleic acid sequence further includes a promoter selected from one of the following: cytomegalovirus immediate early gene promoter (CMV), elongation factor 1α promoter (EF1-α), phosphoglycerate kinase-1 promoter (PGK), ubiquitin-C promoter (UBQ-C), cytomegalovirus enhancer / chicken β-actin promoter (CAG), polyomavirus enhancer / herpes simplex thymidine kinase promoter (MC1), β-actin promoter (β-ACT), simian virus 40 promoter (SV40), myeloproliferative sarcoma virus enhancer, MND promoter, and hypoxia promoter. The sequence of the hypoxia promoter is shown in SEQ ID NO:
62.
4. An expression vector comprising the nucleic acid molecule of claim 2.
5. The expression vector according to claim 4, characterized in that, The expression vector is selected from any one of lentiviral expression vectors, retroviral expression vectors, adenovirus expression vectors, adeno-associated virus expression vectors, RNA vectors, and plasmids.
6. An engineered cell, characterized in that, The cell is transduced with the nucleic acid molecule of claim 2-3 or the expression vector of any one of claim 4-5; the cell is a T cell, a T cell precursor, or an NK cell.
7. A cell product, characterized in that, The cell product includes an engineered cell as described in claim 6.
8. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises the chimeric antigen receptor of claim 1, the nucleic acid molecule of claims 2-3, the expression vector of any one of claims 4-5, an engineered cell of claim 6, or a cell product of claim 7.
9. The use of the chimeric antigen receptor of claim 1, the nucleic acid molecule of claims 2-3, the expression vector of any one of claims 4-5, the engineered cell of claim 6, the cell product of claim 7, or the pharmaceutical composition of claim 8 in the preparation of a drug for treating human B-cell acute lymphoblastic leukemia.