JPWO2020172440A5

JPWO2020172440A5 -

Info

Publication number: JPWO2020172440A5
Application number: JP2021548701A
Authority: JP
Publication date: 2023-02-13

Claims

A nucleic acid molecule comprising a nucleotide sequence encoding a chimeric antigen receptor that targets both BAFF-R and CD19, said chimeric antigen receptor comprising:
A targeting domain, which, from amino-terminus to carboxy-terminus, is:
a) scFv targeting BAFF-R and scFv targeting CD19,
b) scFv targeting CD19 and scFv targeting BAFF-R,
c) VL domain of CD19 scFv, scFv targeting BAFF-R and VH domain of CD19 scFv;
d) VH domain of CD19 scFv, scFv targeting BAFF-R and VL domain of CD19 scFv;
e) the VL domain of BAFF-R scFv, the scFv targeting CD19 and the VH domain of BAFF-R scFv, or
f) comprising the VH domain of BAFF-R scFv, the scFv targeting CD19 and the VL domain of BAFF-R scFv, followed by
spacer domain,
transmembrane domain,
a co-stimulatory domain, and a CD3zeta signaling domain;
A nucleic acid molecule comprising

The VH domain of BAFF-R scFv comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 13-16, and the VL domain of BAFF-R scFv comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 17-20. 2. The nucleic acid molecule of claim 1, comprising:

The VH domain of BAFF-R scFv comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:21-24, and the VL domain of BAFF-R scFv comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:25-28. 3. The nucleic acid molecule of claim 1 or 2 , comprising:

2. The nucleic acid molecule of Claim 1, wherein said chimeric antigen receptor comprises an amino acid sequence selected from SEQ ID NOS:60, 61, 62 and 63.

5. Any of claims 1 to 4 , wherein between said VL domain and scFv and between said VH domain and scFv there is a linker comprising 4-15 amino acids each, said linker comprising only G and S. A nucleic acid molecule according to claim 1.

The co-stimulatory domain is a CD28 co-stimulatory domain or a variant thereof modified by 1-5 amino acids, a 4-1BB co-stimulatory domain or a variant thereof modified by 1-5 amino acids, and an OX40 co-stimulatory domain. A nucleic acid molecule according to any one of claims 1 to 5, selected from the group consisting of a domain or a variant thereof in which 1 to 5 amino acids are modified.

The transmembrane domain is a CD4 transmembrane domain or a variant thereof modified by 1 to 5 amino acids, a CD8 transmembrane domain or a variant thereof modified by 1 to 5 amino acids, a CD28 transmembrane domain or amino acids the CD3ζ transmembrane domain or variants thereof modified by 1-5 amino acids, the co-stimulatory domain or variants thereof modified by 1-5 amino acids; and the CD3ζ signaling domain or variants thereof modified by 1 to 5 amino acids.

The spacer domain is IgG4 hinge (S→P), IgG4 hinge, IgG4 hinge (S228P) + linker, CD28 hinge, CD8 hinge-48aa, CD8 hinge-45aa, IgG4 (HL-CH3), IgG4 (L235E, N297Q) , IgG4 (S228P, L235E, N297Q), and IgG4 (CH3), and IgG4 hinge (S→P), IgG4 hinge, IgG4 hinge (S228P) + linker, CD28 hinge, CD8 hinge-48aa, CD8 hinge-45aa, IgG4 (HL-CH3), IgG4 (L235E, N297Q), IgG4 (S228P, L235E, N297Q), and each variant in which 1 to 5 amino acids of IgG4 (CH3) are modified. , the nucleic acid molecule according to any one of claims 1-7 .

Said BAFF-R scFv comprises a light chain variable region and a heavy chain variable region, said light chain variable region comprising CDR L1 (SEQ ID NO: 1), CDR L2 (SEQ ID NO: 2) and CDR L3 (SEQ ID NO: 3). , said heavy chain variable region comprises CDR H1 (SEQ ID NO:4), CDR H2 (SEQ ID NO:5) and CDR H3 (SEQ ID NO:6), or
The light chain variable region comprises CDR L1 (SEQ ID NO:7), CDR L2 (SEQ ID NO:8) and CDR L3 (SEQ ID NO:9), and the heavy chain variable region comprises CDR H1 (SEQ ID NO:10), CDR H2 (SEQ ID NO:9). No. 11) and CDR H3 (SEQ ID No. 12) .

The BAFF-R scFv comprises a light chain variable domain whose amino acid sequences are selected from SEQ ID NOs: 17-20 and 25-28 , and/or the BAFF-R scFv has the amino acid sequences of SEQ ID NOs: 13-16 and 10. The nucleic acid molecule of any one of claims 1-9 , comprising a heavy chain variable domain selected from 24.

ＣＤ１９ｓｃＦｖは、配列番号３０で表されるアミノ酸配列ＤＩＱＭＴＱＴＴＳＳＬＳＡＳＬＧＤＲＶＴＩＳＣＲＡＳＱＤＩＳＫＹＬＮＷＹＱＱＫＰＤＧＴＶＫＬＬＩＹＨＴＳＲＬＨＳＧＶＰＳＲＦＳＧＳＧＳＧＴＤＹＳＬＴＩＳＮＬＥＱＥＤＩＡＴＹＦＣＱＱＧＮＴＬＰＹＴＦＧＧＧＴＫＬＥＩＴを含むＶＬ、及び、配列番号３１で表されるアミノ酸配列ＥＶＫＬＱＥＳＧＰＧＬＶＡＰＳＱＳＬＳＶＴＣＴＶＳＧＶＳＬＰＤＹＧＶＳＷＩＲＱＰＰＲＫＧＬＥＷＬＧＶＩＷＧＳＥＴＴＹＹＮＳＡＬＫＳＲＬＴＩＩＫＤＮＳＫＳＱＶＦＬＫＭＮＳＬＱＴＤＤＴＡＩＹＹＣＡＫＨＹＹＹＧＧＳＹＡＭＤＹＷＧＱＧＴＳＶＴＶＳＳを含むＶＨを含む、請求項１～１０のいずれか一項に記載の核酸molecule.

A vector or lentiviral vector comprising a nucleic acid molecule according to any one of the preceding claims 1-11 .

13. A population of human T cells or a population of NK cells transduced by the vector or lentiviral vector of claim 12.

14. A composition comprising the population of human T cells of claim 13 for use in a method for treating cancer in a subject by administering to the subject a therapeutically effective amount, wherein the cancer is , lymphoma, leukemia or myeloma .

said lymphoma is mantle cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, marginal zone lymphoma or Burkitt 's lymphoma; or said leukemia is acute lymphoblastic leukemia, chronic lymphocytic leukemia or 15. The composition of claim 14, which is hairy cell leukemia .

16. The composition of claim 14 or 15, wherein said population of T cells is autologous or allogeneic to said patient, or said human T cell population comprises CD4+ and CD8+ cells .