NZ740714B2

NZ740714B2 - T cell receptors recognizing hla-cw8 restricted mutated kras

Info

Publication number: NZ740714B2
Application number: NZ740714A
Authority: NZ
Inventors: Yong Chen Lu; Steven A Rosenberg; Eric Tran
Original assignee: The United States Of America As Represented By The Secretary Department Of Health And Human Services
Filing date: 2016-09-09
Publication date: 2025-05-01

Abstract

Disclosed is an isolated or purified T cell receptor (TCR) having antigenic specificity for mutated Kirsten rat sarcoma viral oncogene homolog (KRAS) presented in the context of an HLA-Cw*0802 molecule. Related polypeptides and proteins, as well as related nucleic acids, recombinant expression vectors, host cells, populations of cells, and pharmaceutical compositions are also provided. Also disclosed are methods of detecting the presence of cancer in a mammal and methods of treating or preventing cancer in a mammal.

Claims

CLAIM (S):

1. An isolated or purified T cell receptor (TCR) comprising: (I) an a chain complementarity determining region (CDR) 1 comprising the amino acid sequence of SEQ ID NO: 3, an a chain CDR2 comprising the amino acid sequence of SEQ ID NO: 4, and an a chain CDR3 comprising the amino acid sequence of SEQ ID NO: 5; (II) a ß chain CDR1 comprising the amino acid sequence of SEQ ID NO: 6, a ß chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a ß chain CDR3 comprising the amino acid sequence of SEQ ID NO: 8.

2. The isolated or purified TCR of claim 1, wherein the TCR has antigenic specificity for the mutated KRAS amino acid sequence of GADGVGKSA (SEQ ID NO: 18) presented by a human leukocyte antigen (HLA)-Cw8 molecule.

3. The isolated or purified TCR of claim 2, wherein the HLA-Cw8 molecule is HLA- Cw*0802.

4. The isolated or purified TCR of any one of claims 1-3, comprising: (I) an a chain variable region comprising an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 9; (II) a ß chain variable region comprising an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 10; (III) an a chain variable region comprising an amino acid sequence at least 99% identical to amino acids 20-129 of SEQ ID NO: 9; (IV) a ß chain variable region comprising an amino acid sequence at least 99% identical to amino acids 22-132 of SEQ ID NO: 10; or (V) both (I) and (II), both (I) and (IV), both (II) and (III), or both (III) and (IV).

5. The isolated or purified TCR of any one of claims 1-4, comprising: (I) an a chain variable region comprising the amino acid sequence of SEQ ID NO: 9; (II) a ß chain variable region comprising the amino acid sequence of SEQ ID NO: 10; (III) an a chain variable region comprising amino acids 20-129 of SEQ ID NO: 9; (IV) a ß chain variable region comprising amino acids 22-132 of SEQ ID NO: 10; or (V) both (I) and (II), both (I) and (IV), both (II) and (III), or both (III) and (IV).

6. The isolated or purified TCR of any one of claims 1-5, further comprising: (a) an a chain constant region comprising an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 11, wherein: (i) X at position 48 of SEQ ID NO: 11 is Thr or Cys; (ii) X at position 112 of SEQ ID NO: 11 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 114 of SEQ ID NO: 11 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; (iv) X at position 115 of SEQ ID NO: 11 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (b) a ß chain constant region comprising an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 12, wherein X at position 57 of SEQ ID NO: 12 is Ser or Cys; or (c) both (a) and (b).

7. The isolated or purified TCR of any one of claims 1-5, further comprising: (a) an a chain constant region comprising the amino acid sequence of SEQ ID NO: 11, wherein: (i) X at position 48 of SEQ ID NO: 11 is Thr or Cys; (ii) X at position 112 of SEQ ID NO: 11 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 114 of SEQ ID NO: 11 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; (iv) X at position 115 of SEQ ID NO: 11 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (b) a ß chain constant region comprising the amino acid sequence of SEQ ID NO: 12, wherein X at position 57 of SEQ ID NO: 12 is Ser or Cys; or (c) both (a) and (b).

8. The isolated or purified TCR of any one of claims 1-7, comprising: (a) an a chain comprising an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 13, wherein: (i) X at position 177 of SEQ ID NO: 13 is Thr or Cys; (ii) X at position 241 of SEQ ID NO: 13 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 243 of SEQ ID NO: 13 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; (iv) X at position 244 of SEQ ID NO: 13 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (b) a ß chain comprising an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 14, wherein X at position 189 of SEQ ID NO: 14 is Ser or (c) an a chain comprising an amino acid sequence at least 99% identical to amino acids 20-266 of SEQ ID NO: 13, wherein: (i) X at position 177 of SEQ ID NO: 13 is Thr or Cys; (ii) X at position 241 of SEQ ID NO: 13 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 243 of SEQ ID NO: 13 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; (iv) X at position 244 of SEQ ID NO: 13 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (d) a ß chain comprising an amino acid sequence at least 99% identical to amino acids 22-305 of SEQ ID NO: 14, wherein X at position 189 of SEQ ID NO: 14 is Ser or Cys; or (e) both (a) and (b), both (a) and (d), both (b) and (c), or both (c) and (d).

9. The isolated or purified TCR of any one of claims 1-7, comprising: (a) an a chain comprising the amino acid sequence of SEQ ID NO: 13, wherein: (i) X at position 177 of SEQ ID NO: 13 is Thr or Cys; (ii) X at position 241 of SEQ ID NO: 13 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 243 of SEQ ID NO: 13 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; (iv) X at position 244 of SEQ ID NO: 13 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (b) a ß chain comprising the amino acid sequence of SEQ ID NO: 14, wherein X at position 189 of SEQ ID NO: 14 is Ser or Cys; or (c) an a chain comprising amino acids 20-266 of SEQ ID NO: 13, wherein: (i) X at position 177 of SEQ ID NO: 13 is Thr or Cys; (ii) X at position 241 of SEQ ID NO: 13 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 243 of SEQ ID NO: 13 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; (iv) X at position 244 of SEQ ID NO: 13 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (d) a ß chain comprising amino acids 22-305 of SEQ ID NO: 14, wherein X at position 189 of SEQ ID NO: 14 is Ser or Cys; or (e) both (a) and (b), both (a) and (d), both (b) and (c), or both (c) and (d).

10. An isolated or purified polypeptide comprising a functional portion of the TCR of any one of claims 1-9, wherein the functional portion comprises the amino acid sequences of SEQ ID NOs: 3-8.

11. The isolated or purified polypeptide of claim 10, wherein the functional portion comprises amino acid sequence(s): (I) at least 99% identical to the amino acid sequence of SEQ ID NO: 9; (II) at least 99% identical to the amino acid sequence of SEQ ID NO: 10; (III) at least 99% identical to amino acids 20-129 of SEQ ID NO: 9; (IV) at least 99% identical to amino acids 22-132 of SEQ ID NO: 10; or (V) both (I) and (II); both (I) and IV); both (II) and (III); or both (III) and (IV).

12. The isolated or purified polypeptide of claim 10, wherein the functional portion comprises the amino acid sequences of (I) SEQ ID NO: 9; (II) SEQ ID NO: 10; (III) amino acids 20-129 of SEQ ID NO: 9; (IV) amino acids 22-132 of SEQ ID NO: 10; or (V) both (I) and (II); both (I) and (IV); both (II) and (III); or both (III) and (IV).

13. The isolated or purified polypeptide of any one of claims 10-12, further comprising: (a) an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 11, wherein: (i) X at position 48 of SEQ ID NO: 11 is Thr or Cys; (ii) X at position 112 of SEQ ID NO: 11 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 114 of SEQ ID NO: 11 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; (iv) X at position 115 of SEQ ID NO: 11 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (b) an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 12, wherein X at position 57 of SEQ ID NO: 12 is Ser or Cys; or (c) both (a) and (b).

14. The isolated or purified polypeptide of any one of claims 10-12, further comprising: (a) the amino acid sequence of SEQ ID NO: 11, wherein: (i) X at position 48 of SEQ ID NO: 11 is Thr or Cys; (ii) X at position 112 of SEQ ID NO: 11 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 114 of SEQ ID NO: 11 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; (iv) X at position 115 of SEQ ID NO: 11 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (b) the amino acid sequence of SEQ ID NO: 12, wherein X at position 57 of SEQ ID NO: 12 is Ser or Cys; or (c) both (a) and (b).

15. The isolated or purified polypeptide of any one of claims 10-14, comprising: (a) an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 13, wherein: (i) X at position 177 of SEQ ID NO: 13 is Thr or Cys; (ii) X at position 241 of SEQ ID NO: 13 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 243 of SEQ ID NO: 13 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; (iv) X at position 244 of SEQ ID NO: 13 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (b) an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 14, wherein X at position 189 of SEQ ID NO: 14 is Ser or Cys; (c) an amino acid sequence at least 99% identical to amino acids 20-266 of SEQ ID NO: 13, wherein: (i) X at position 177 of SEQ ID NO: 13 is Thr or Cys; (ii) X at position 241 of SEQ ID NO: 13 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 243 of SEQ ID NO: 13 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; (iv) X at position 244 of SEQ ID NO: 13 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (d) an amino acid sequence at least 99% identical to amino acids 22-305 of SEQ ID NO: 14, wherein X at position 189 of SEQ ID NO: 14 is Ser or Cys; or (e) both (a) and (b), both (a) and (d), both (b) and (c), or both (c) and (d).

16. The isolated or purified polypeptide of any one of claims 10-14, comprising: (a) the amino acid sequence of SEQ ID NO: 13, wherein: (i) X at position 177 of SEQ ID NO: 13 is Thr or Cys; (ii) X at position 241 of SEQ ID NO: 13 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 243 of SEQ ID NO: 13 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; (iv) X at position 244 of SEQ ID NO: 13 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (b) the amino acid sequence of SEQ ID NO: 14, wherein X at position 189 of SEQ ID NO: 14 is Ser or Cys; (c) amino acids 20-266 of SEQ ID NO: 13, wherein: (i) X at position 177 of SEQ ID NO: 13 is Thr or Cys; (ii) X at position 241 of SEQ ID NO: 13 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 243 of SEQ ID NO: 13 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; (iv) X at position 244 of SEQ ID NO: 13 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (d) amino acids 22-305 of SEQ ID NO: 14, wherein X at position 189 of SEQ ID NO: 14 is Ser or Cys; or (e) both (a) and (b), both (a) and (d), both (b) and (c), or both (c) and (d).

17. An isolated or purified protein comprising a functional portion of the TCR of any one of claims 1-9, wherein the functional portion comprises the amino acid sequences of SEQ ID NOs: 3-8, and wherein the protein comprises a first polypeptide chain comprising the amino acid sequences of SEQ ID NOs: 3-5 and a second polypeptide chain comprising the amino acid sequences of SEQ ID NOs: 6-8.

18. The isolated or purified protein of claim 17, wherein: (I) the first polypeptide chain comprises an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 9; (II) the second polypeptide chain comprises an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 10; (III) the first polypeptide chain comprises an amino acid sequence at least 99% identical to amino acids 20-129 of SEQ ID NO: 9; (IV) the second polypeptide chain comprises an amino acid sequence at least 99% identical to amino acids 22-132 of SEQ ID NO: 10; or (V) both (I) and (II), both (I) and (IV), both (II) and (III), or both (III) and (IV).

19. The isolated or purified protein of claim 17, wherein: (I) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 9; (II) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: (III) the first polypeptide chain comprises amino acids 20-129 of SEQ ID NO: 9; (IV) the second polypeptide chain comprises amino acids 22-132 of SEQ ID NO: 10; (V) both (I) and (II), both (I) and (IV), both (II) and (III), or both (III) and (IV).

20. The isolated or purified protein of any one of claims 17-19, wherein: (a) the first polypeptide chain comprises an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 11, wherein: (i) X at position 48 of SEQ ID NO: 11 is Thr or Cys; (ii) X at position 112 of SEQ ID NO: 11 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 114 of SEQ ID NO: 11 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; (iv) X at position 115 of SEQ ID NO: 11 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (b) the second polypeptide chain comprises an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 12, wherein X at position 57 of SEQ ID NO: 12 is Ser or Cys; or (c) both (a) and (b).

21. The isolated or purified protein of any one of claims 17-19, wherein: (a) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 11, wherein: (i) X at position 48 of SEQ ID NO: 11 is Thr or Cys; (ii) X at position 112 of SEQ ID NO: 11 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 114 of SEQ ID NO: 11 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; (iv) X at position 115 of SEQ ID NO: 11 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (b) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 12, wherein X at position 57 of SEQ ID NO: 12 is Ser or Cys; or (c) both (a) and (b).

22. The isolated or purified protein of any one of claims 17-21, wherein: (a) the first polypeptide chain comprises an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 13, wherein: (i) X at position 177 of SEQ ID NO: 13 is Thr or Cys; (ii) X at position 241 of SEQ ID NO: 13 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 243 of SEQ ID NO: 13 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; (iv) X at position 244 of SEQ ID NO: 13 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (b) the second polypeptide chain comprises an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 14, wherein X at position 189 of SEQ ID NO: 14 is Ser or Cys; (c) the first polypeptide chain comprises an amino acid sequence at least 99% identical to amino acids 20-266 of SEQ ID NO: 13, wherein: (i) X at position 177 of SEQ ID NO: 13 is Thr or Cys; (ii) X at position 241 of SEQ ID NO: 13 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 243 of SEQ ID NO: 13 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; (iv) X at position 244 of SEQ ID NO: 13 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (d) the second polypeptide chain comprises an amino acid sequence at least 99% identical to amino acids 22-305 of SEQ ID NO: 14, wherein X at position 189 of SEQ ID NO: 14 is Ser or Cys; or (e) both (a) and (b), both (a) and (d), both (b) and (c), or both (c) and (d).

23. The isolated or purified protein of any one of claims 17-21, wherein: (a) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 13, wherein: (i) X at position 177 of SEQ ID NO: 13 is Thr or Cys; (ii) X at position 241 of SEQ ID NO: 13 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 243 of SEQ ID NO: 13 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; (iv) X at position 244 of SEQ ID NO: 13 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (b) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 14, wherein X at position 189 of SEQ ID NO: 14 is Ser or Cys; or (c) the first polypeptide chain comprises amino acids 20-266 of SEQ ID NO: 13, wherein: (i) X at position 177 of SEQ ID NO: 13 is Thr or Cys; (ii) X at position 241 of SEQ ID NO: 13 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 243 of SEQ ID NO: 13 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; (iv) X at position 244 of SEQ ID NO: 13 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (d) the second polypeptide chain comprises amino acids 22-305 of SEQ ID NO: 14, wherein X at position 189 of SEQ ID NO: 14 is Ser or Cys; or (e) both (a) and (b), both (a) and (d), both (b) and (c), or both (c) and (d).

24. An isolated or purified nucleic acid which encodes the TCR of any one of claims 1-9, the polypeptide of any one of claims 10-16, or the protein of any one of claims 17-23.

25. A recombinant expression vector comprising the nucleic acid of claim 24.

26. An isolated or purified host cell comprising the recombinant expression vector of claim 25.

27. An isolated or purified population of cells comprising at least two isolated or purified host cells of claim 26.

28. A pharmaceutical composition comprising the TCR of any one of claims 1-9, the polypeptide of any one of claims 10-16, the protein of any one of claims 17-23, the nucleic acid of claim 24, the recombinant expression vector of claim 25, the host cell of claim 26, or the population of host cells of claim 27, and a pharmaceutically acceptable carrier.

29. A method of detecting the presence of cancer in a mammal, the method comprising: (a) contacting a sample obtained from said mammal, wherein said sample comprises cells of the cancer, with the TCR of any one of claims 1-9, the polypeptide of any one of claims 10-16, the protein of any one of claims 17-23, the nucleic acid of claim 24, the recombinant expression vector of claim 25, the host cell of claim 26, the population of host cells of claim 27, or the pharmaceutical composition of claim 28, thereby forming a complex; (b) detecting the complex, wherein detection of the complex is indicative of the presence of cancer in the mammal.

30. The method of claim 29, wherein the cancer is pancreatic, colorectal, lung, endometrial, ovarian, or prostate cancer.

31. Use of the TCR of any one of claims 1-9, the polypeptide of any one of claims 10-16, the protein of any one of claims 17-23, the nucleic acid of claim 24, the recombinant expression vector of claim 25, the host cell of claim 26, the population of host cells of claim 27, or the pharmaceutical composition of claim 28, in the manufacture of a medicament for the treatment or prevention of cancer in a human.

32. The use of claim 31, wherein the cancer is pancreatic, colorectal, lung, endometrial, ovarian, or prostate cancer.

33. An isolated or purified T cell receptor (TCR) as claimed in any one of claims 1-9; an isolated or purified polypeptide as claimed in any one of claims 10-16; an isolated or purified protein as claimed in any one of claims 17-23; an isolated or purified nucleic acid as claimed in claim 24; a recombinant expression vector as claimed in claim 25; an isolated or purified host cell as claimed in claim 26; an isolated or purified population of cells as claimed in claim 27; a pharmaceutical composition as claimed in claim 28; a method as claimed in claim 29 or 20; or a use as claimed in claim 31 or 32, substantially as described herein and with reference to any example thereof.