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US 2026/0265336 A1Published application
High-affinity tcr for identifying mage-a4 antigen, and sequence and use thereof
AssigneeXlifesc, Ltd.
InventorsKai Zhan, Jinhua Huang, Wenjing Zheng, Qianxia Yin, Xianqing Tang
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Abstract
Provided are a high-affinity TCR for identifying a MAGE-A4 antigen, and a sequence and a use thereof. The TCR has the characteristic of binding to a GVYDGREHTV-HLA A0201 complex. Also provided are a multivalent TCR complex, a nucleic acid molecule encoding the TCR, a vector comprising the nucleic acid molecule, a cell expressing the TCR, and a pharmaceutical composition comprising the substance, and a preparation method therefor.
Claims21
1:
- A T cell receptor (TCR), comprising a TCR α chain variable domain and a TCR β chain variable domain, wherein the TCR has an activity of binding to a GVYDGREHTV-HLA A0201 complex;
- an amino acid sequence of the TCR α chain variable domain has at least 90% sequence homology to an amino acid sequence shown in SEQ ID NO:1, and an amino acid sequence of the TCR β chain variable domain has at least 90% sequence homology to an amino acid sequence shown in SEQ ID NO:2.
2:
- The TCR according to claim 1, wherein three complementarity determining regions (CDRs) of the TCR β chain variable domain are:
- CDR1β:
- MNHEY, CDR2β:
- SVGEGT, and CDR 3β:
- ASSLGRAYEQY;
- preferably, the amino acid sequence of the TCR β chain variable domain is SEQ ID NO:2.
3 more elements
3:
- The TCR according to claim 1, wherein the amino acid sequence of the TCR α chain variable domain has at least 95% sequence homology to the amino acid sequence shown in SEQ ID NO:1, and the amino acid sequence of the TCR β chain variable domain has at least 95% sequence homology to the amino acid sequence shown in SEQ ID NO:2.
4:
- The TCR according to claim 1, wherein the affinity of the TCR to the GVYDGREHTV-HLA A0201 complex is at least 5 times that of a wild-type TCR.
5:
- The TCR according to claim 1, wherein reference sequences of three CDRs (complementarity-determining regions) of the TCR α chain variable domain are as follows:
- CDR1α:
- DSSSTY,
- CDR2α:
- IFSNMDM, and
- CDR3α:
- AEQSFGNEKLT;
- CDR3α contains at least one of the following mutations:
- Residue before mutation
- Residue after mutation
- E at position 2 of CDR3α
- W, V, A, T, S, H, Q, N, or Y
- Q at position 3 of CDR3α
- H or S
- S at position 4 of CDR3α
- T, Q, N, E, or D
- F at position 5 of CDR3α
- K, R, or H
- G at position 6 of CDR3α
- Q
- preferably, an amino acid mutation in CDR3α contains:
- Residue before mutation
- Residue after mutation
- Q at position 3 of CDR3α
- S
22 more elements
6:
- The TCR according to claim 1, wherein the TCR has CDRs selected from the following groups:
- CDR number
- α-CDR1
- α-CDR2
- α-CDR3
- ß-CDR1
- ß-CDR2
- ß-CDR3
- 4
- DSSSTY
- IFSNMDM
- AASEFGNEKLT
- MNHEY
- SVGEGT
- ASSLGRAYEQY
- 2
- DSSSTY
- IFSNMDM
- AWSQFGNEKLT
- MNHEY
- SVGEGT
- ASSLGRAYEQY
- 1
- DSSSTY
- IFSNMDM
- AEHTKQNEKLT
- MNHEY
- SVGEGT
- ASSLGRAYEQY
- 3
- DSSSTY
- IFSNMDM
- AVSNFGNEKLT
- MNHEY
- SVGEGT
- ASSLGRAYEQY
- 5
- DSSSTY
- IFSNMDM
- AASTFGNEKLT
- MNHEY
- SVGEGT
- ASSLGRAYEQY
- 6
- DSSSTY
- IFSNMDM
- AASQFGNEKLT
- MNHEY
- SVGEGT
- ASSLGRAYEQY
- 7
- DSSSTY
- IFSNMDM
- AASQRGNEKLT
- MNHEY
- SVGEGT
- ASSLGRAYEQY
- 8
- DSSSTY
- IFSNMDM
- AVSQFGNEKLT
- MNHEY
- SVGEGT
- ASSLGRAYEQY
- 9
- DSSSTY
- IFSNMDM
- AVSDFGNEKLT
- MNHEY
- SVGEGT
- ASSLGRAYEQY
- 10
- DSSSTY
- IFSNMDM
- AVSSFGNEKLT
- MNHEY
- SVGEGT
- ASSLGRAYEQY
- 11
- DSSSTY
- IFSNMDM
- ATSTFGNEKLT
- MNHEY
- SVGEGT
- ASSLGRAYEQY
- 12
- DSSSTY
- IFSNMDM
- ATSQFGNEKLT
- MNHEY
- SVGEGT
- ASSLGRAYEQY
- 13
- DSSSTY
- IFSNMDM
- ATSDFGNEKLT
- MNHEY
- SVGEGT
- ASSLGRAYEQY
- 14
- DSSSTY
- IFSNMDM
- ASSQFGNEKLT
- MNHEY
- SVGEGT
- ASSLGRAYEQY
- 15
- DSSSTY
- IFSNMDM
- AWSSFGNEKLT
- MNHEY
- SVGEGT
- ASSLGRAYEQY
- 16
- DSSSTY
- IFSNMDM
- AHSQFGNEKLT
- MNHEY
- SVGEGT
- ASSLGRAYEQY
- 17
- DSSSTY
- IFSNMDM
- AQSQFGNEKLT
- MNHEY
- SVGEGT
- ASSLGRAYEQY
- 18
- DSSSTY
- IFSNMDM
- ANSTFGNEKLT
- MNHEY
- SVGEGT
- ASSLGRAYEQY
- 19
- DSSSTY
- IFSNMDM
- AYSQFGNEKLT
- MNHEY
- SVGEGT
- ASSLGRAYEQY
- 20
- DSSSTY
- IFSNMDM
- AEHTHQNEKLT
- MNHEY
- SVGEGT
- ASSLGRAYEQY
145 more elements
7:
- The TCR according to claim 1, wherein the TCR is soluble;
- preferably, the TCR contains (i) the TCR α chain variable domain and all or a part of a TCR α chain constant region except a transmembrane domain;
- and (ii) the TCR β chain variable domain and all or a part of a TCR β chain constant region except a transmembrane domain.
8:
- The TCR according to claim 1, wherein the TCR is an αβ heterodimeric TCR, containing an α chain TRAC constant region sequence and a β chain TRBC1 or TRBC2 constant region sequence.
9:
- The TCR according to claim 1, wherein the TCR contains an α chain constant region and a β chain constant region;
- an artificial interchain disulfide bond is contained between the α chain constant region and the β chain constant region;
- preferably, cysteine residues forming the artificial interchain disulfide bond between the α chain constant region and the β chain constant region of the TCR replace one or more of the following groups of sites:
- Thr48 of exon 1 of TRAC*01 and Ser57 of exon 1 of TRBC1*01 or TRBC2*01;
- Thr45 of exon 1 of TRAC*01 and Ser77 of exon 1 of TRBC1*01 or TRBC2*01;
- Tyr10 of exon 1 of TRAC*01 and Ser17 of exon 1 of TRBC1*01 or TRBC2*01;
- Thr45 of exon 1 of TRAC*01 and Asp59 of exon 1 of TRBC1*01 or TRBC2*01;
- Ser15 of exon 1 of TRAC*01 and Glu15 of exon 1 of TRBC1*01 or TRBC2*01;
- Arg53 of exon 1 of TRAC*01 and Ser54 of exon 1 of TRBC1*01 or TRBC2*01;
- Pro89 of exon 1 of TRAC*01 and Ala19 of exon 1 of TRBC1*01 or TRBC2*01;
- and
- Tyr10 of exon 1 of TRAC*01 and Glu20 of exon 1 of TRBC1*01 or TRBC2*01.
9 more elements
10:
- The TCR according to claim 1, wherein the amino acid sequence of the TCR α chain variable domain is one of SEQ ID NO:16, 14, 13, 15, or 17-32;
- and the amino acid sequence of the TCR β chain variable domain is SEQ ID NO:
- 2.
11:
- The TCR according to claim 1, wherein the TCR is a single-chain TCR;
- preferably, the TCR is a single-chain TCR comprising the α chain variable domain and the β chain variable domain;
- the α chain variable domain and the β chain variable domain are linked by a flexible short peptide sequence (linker).
12:
- The TCR according to claim 1, wherein a C- or N-terminus of the α chain and/or β chain of the TCR is bound with a conjugate;
- the conjugate is a detectable marker or a therapeutic agent;
- or
- the therapeutic agent that binds to the TCR is an anti-CD3 antibody linked to the C- or N-terminus of the α chain or β chain of the TCR.
1 more element
13:
- A multivalent T cell receptor (TCR) complex, containing at least two TCR molecules, wherein at least one TCR molecule is the TCR according to claim 1.
14:
- A nucleic acid molecule, containing a nucleic acid sequence encoding the TCR according to claim 1 or a complementary sequence of the nucleic acid sequence.
15:
- A vector, containing the nucleic acid molecule according to claim 14.
16:
- A host cell, containing a vector containing a nucleic acid sequence encoding the TCR according to claim 1 or a complementary sequence of the nucleic acid sequence or having a chromosome integrated with an exogenous nucleic acid molecule containing a nucleic acid sequence encoding the TCR according to claim 1 or a complementary sequence of the nucleic acid sequence.
17:
- An isolated cell, expressing the T cell receptor (TCR) according to claim 1, wherein the isolated cell is a T cell.
18:
- A pharmaceutical composition, containing the T cell receptor (TCR) according to claim 1, the TCR complex containing at least two TCR molecules, wherein at least one TCR molecule is the TCR according to claim 1, or an isolated cell expressing the T cell receptor (TCR) according to claim 1, wherein the isolated cell is a T cell.
19:
- A method for treating a disease, comprising:
- administering the T cell receptor (TCR) according to claim 1, the TCR complex containing at least two TCR molecules, wherein at least one TCR molecule is the TCR according to claim 1, an isolated cell expressing the T cell receptor (TCR) according to claim 1, wherein the isolated cell is a T cell, or a pharmaceutical composition containing the T cell receptor (TCR) according to claim 1, the TCR complex containing at least two TCR molecules, wherein at least one TCR molecule is the TCR according to claim 1, or an isolated cell expressing the T cell receptor (TCR) according to claim 1, wherein the isolated cell is a T cell to a subject in need of treatment, wherein the disease is a MAGE-A4 positive tumor.
20:
- A method for preparing the T cell receptor (TCR) according to claim 1, comprising the steps of:
- (i) culturing an isolated cell expressing the T cell receptor (TCR) according to claim 1, wherein the isolated cell is a T cell to express the TCR according to claim 1;
- and
- (ii) obtaining the TCR through isolating or purifying.
1 more element
21-26. (canceled)