NZ747974B2

NZ747974B2 - T cell receptors

Info

Publication number: NZ747974B2
Application number: NZ747974A
Authority: NZ
Inventors: Eleanor Bagg; William Lawrance; Nicholas Tribble
Original assignee: Adaptimmune Limited
Filing date: 2017-04-10
Publication date: 2025-03-21

Abstract

The present invention relates to T cell receptors (TCRs) which bind the HLA-A*0201 restricted peptide GVYDGREHTV (SEQ ID NO: 1) derived from the MAGE-A4 protein. The TCRs of the invention demonstrate excellent specificity profiles for this MAGE epitope. Also provided are nucleic acids encoding the TCRs, cells engineered to present the TCRs, cells harbouring expression vectors encoding the TCRs and pharmaceutical compositions comprising the TCRs, nucleic acids or cells of the invention.

Claims

The Claims Defining the Invention are as Follows:

1. A T cell receptor (TCR) that comprises a TCR alpha chain variable domain and a TCR beta chain variable domain, wherein: Alpha CDR1 is VSPFSN; 5 Alpha CDR2 is LTIMTF or LTIVTF or LTILTF; Alpha CDR3 is CVVSGGTDSWGKLQF; Beta CDR1 is KGHDR; Beta CDR2 is SFDVK; and Beta CDR3 is CATSGQGAYEEQFF. 10

2. A TCR according to claim 1, wherein the TCR has the property of binding to GVYDGREHTV (SEQ ID No: 1) in complex with HLA-A*0201 with a dissociation constant of from 0.05 µM to 20.0 µM when measured with surface plasmon resonance at 25ºC and at a pH between 7.1 and 7.5 using a soluble form of the TCR, and has at least a ten-fold selectivity of binding to SEQ ID No:1 in complex with HLA-A*0201 over binding to GVYDGEEHSV (SEQ ID No 2) in 15 complex with HLA-A*0201, and wherein the TCR variable domains form contacts with at least residues V2, Y3 and D4 of GVYDGREHTV (SEQ ID No: 1).

3. A TCR according to claim 1 or 2, which is an alpha-beta heterodimer, having an alpha chain TRAV10 + TRAC constant domain sequence and a beta chain TRBV24-1 + TRBC-2 constant domain sequence. 20 4. A TCR as claimed in claim 1 or 2, which is in single chain format of the type Va-L-Vß, Vß-

4. L-Va, Va-Ca-L-Vß, or Va-L-Vß-Cß, wherein Va and Vß are TCR a and ß variable regions respectively, Ca and Cß are TCR a and ß constant regions respectively, and L is a linker sequence.

5. A TCR as claimed in any one of the preceding claims, which is covalently associated with a 25 detectable label, a therapeutic agent or a PK modifying moiety.

6. A TCR as claimed in any one of the preceding claims, wherein the TCR alpha chain variable domain comprises an amino acid sequence that has at least 80% identity to the sequence of amino acid residues 1-105 of SEQ ID No: 3 and wherein Alpha CDR2 is LTIVTF or LTILTF, and/or the TCR beta chain variable domain comprises an amino acid sequence that has at least 30 80% identity to the sequence of amino acid residues 1-123 of SEQ ID No: 4.

7. A TCR as claimed in any one of the preceding claims, wherein the TCR alpha chain variable domain is comprised in a sequence in which: (i) amino acid residues 1-27 thereof has (a) at least 90% identity to the sequence of amino acid residues 1-27 of SEQ ID No: 3 or (b) one, two or three amino acid insertions, deletions or 35 substitutions relative to the sequence of (a); (ii) amino acid residues 28-33 is VSPFSN; (iii) amino acid residues 34-47 thereof has (a) at least 90% identity to the sequence of amino acid residues 34-47 of SEQ ID NO: 3 or (b) one, two or three amino acid insertions, deletions or substitutions relative to the sequence of (a); 5 (iv) amino acid residues 48-53 is LTIMTF or LTIVTF or LTILTF; (v) amino acid residues 54-90 thereof has (a) at least 90% identity to the sequence of amino acid residues 54-90 of SEQ ID No: 3 or (b) one, two or three amino acid insertions, deletions or substitutions relative to the sequence of (a); and (vi) amino acid residues 91-105 is CVVSGGTDSWGKLQF. 10 8. A TCR as claimed in any one of the preceding claims, wherein the TCR alpha chain variable domain is comprised in the amino acid sequence of SEQ ID No: 3 or 7 or

8.

9. A TCR as claimed in any one of the preceding claims, wherein the TCR alpha chain variable domain is comprised in the amino acid sequence of SEQ ID No: 7 or 8.

10. A TCR according to any one of the preceding claims, wherein the TCR beta chain variable 15 domain is comprised in a sequence in which: (i) amino acid residues 1-45 thereof has (a) at least 90% identity to the sequence of amino acid residues 1-45 of SEQ ID No: 4 or (b) one, two or three amino acid insertions, deletions or substitutions relative to the sequence of (a); (ii) amino acid residues 46-50 is KGHDR; 20 (iii) amino acid residues 51-67 thereof has (a) at least 90% identity to the sequence of amino acid residues 51-67 of SEQ ID NO: 4 or (b) one, two or three amino acid insertions, deletions or substitutions to the sequence of (a); (iv) amino acid residues 68-72 is SFDVK; (v) amino acid residues 73-109 thereof has (a) at least 90% identity to the sequence of amino 25 acid residues 73-109 of SEQ ID NO: 4 or (b) one, two or three amino acid insertions, deletions or substitutions relative to the sequence of (a); and (vi) amino acid residues 110-123 is CATSGQGAYEEQFF.

11. A TCR as claimed in any one of the preceding claims, wherein the TCR beta chain variable domain is comprised in the amino acid sequence of SEQ ID No: 9.

12. A TCR as claimed in any one of the preceding claims, wherein the TCR alpha chain variable domain is comprised in the amino acid sequence of any one of SEQ ID Nos: 3, 7 and 8, and the TCR beta chain variable domain is comprised in the amino acid sequence of SEQ ID

13. No: 9. 5 13. A TCR as claimed in any one of the preceding claims, wherein the TCR alpha chain variable domain is comprised in the amino acid sequence of SEQ ID No: 8, and the TCR beta chain variable domain is comprised in the amino acid sequence of SEQ ID No: 9.

14. Nucleic acid encoding a TCR as claimed in any one of the preceding claims.

15. An isolated cell, presenting a TCR as claimed in any one of claims 1 to 13. 10

16. The isolated cell as claimed in claim 15, wherein the cell is a T-cell.

17. An isolated cell presenting a TCR encoded by a nucleic acid, wherein the nucleic acid encodes a TCR that comprises: (A) a TCR alpha chain variable domain in which: (i) amino acid residues 1-27 thereof has (a) at least 90% identity to the sequence of amino 15 acid residues 1-27 of SEQ ID No: 3 or (b) one, two or three amino acid insertions, deletions or substitutions relative to the sequence of (a); (ii) amino acid residues 28-33 is VSPFSN; (iii) amino acid residues 34-47 thereof has (a) at least 90% identity to the sequence of amino acid residues 34-47 of SEQ ID NO: 3 or (b) one, two or three amino acid insertions, 20 deletions or substitutions relative to the sequence of (a); (iv) amino acid residues 48-53 is LTIMTF or LTIVTF or LTILTF; (v) amino acid residues 54-90 thereof has (a) at least 90% identity to the sequence of amino acid residues 54-90 of SEQ ID No: 3 or (b) one, two or three amino acid insertions, deletions or substitutions relative to the sequence of (a); and 25 (vi) amino acid residues 91-105 is CVVSGGTDSWGKLQF; and (B) a TCR beta chain variable domain in which: (i) amino acid residues 1-45 thereof has (a) at least 90% identity to the sequence of amino acid residues 1-45 of SEQ ID No: 4 or (b) one, two or three amino acid insertions, deletions or substitutions relative to the sequence of (a); 30 (ii) amino acid residues 46-50 is KGHDR; (iii) amino acid residues 51-67 thereof has (a) at least 90% identity to the sequence of amino acid residues 51-67 of SEQ ID NO: 4 or (b) one, two or three amino acid insertions, deletions or substitutions relative to the sequence of (a); (iv) amino acid residues 68-72 is SFDVK; 35 (v) amino acid residues 73-109 thereof has (a) at least 90% identity to the sequence of amino acid residues 73-109 of SEQ ID NO: 4 or (b) one, two or three amino acid insertions, deletions or substitutions relative to the sequence of (a); and (vi) amino acid residues 110-123 is CATSGQGAYEEQFF.

18. An isolated cell harbouring: (a) a TCR expression vector which comprises nucleic acid as claimed in claim 14 in a single open reading frame, or two distinct open reading frames encoding the alpha chain and the 5 beta chain respectively; or (b) a first expression vector which comprises nucleic acid encoding the alpha chain of a TCR as claimed in any one of claims 1 to 13, and a second expression vector which comprises nucleic acid encoding the beta chain of a TCR as claimed in any one of claims 1 to 13.

19. A pharmaceutical composition comprising a TCR as claimed in any one of claims 1 to 13, 10 nucleic acid of claim 14 or a cell as claimed in any one of claims 15 to 18, together with one or more pharmaceutically acceptable carriers or excipients.

20. The TCR of any one of claims 1 to 13, nucleic acid of claim 14 or cell of any one of claims 15 to 18 for use in medicine.

21. Use of the TCR of any one of claims 1 to 13, the nucleic acid of claim 14, or the cell of any 15 one of claims 15 to 18, in the manufacture of a medicament for the treatment of cancer.