Interleukin-21 mutein/ anti-PD-1 antibody conjugates
Provided herein are IL-21 muteins and fusion proteins comprising the same for use in methods of treating a disease. Related conjugates, nucleic acids, vectors, host cells, pharmaceutical compositions and kits are also provided herein. Methods of making the IL-21 muteins and fusion proteins comprising the same, as well as methods of treating a subject in need thereof, are provided by the present disclosure. Further provided are PD-1 antigen-binding proteins.
1. A conjugate comprising:
an IL-21 mutein comprising an amino acid sequence selected from SEQ ID NOs: 233-245; and
an anti-PD-1 antibody comprising:
two light chains, each comprising an LC CDR1, LC CDR2, and LC CDR3 comprising the amino acid sequences of SEQ ID NOs: 385, 386, and 387, respectively; and
two heavy chains, each comprising an HC CDR1, HC CDR2, and HC CDR3 comprising the amino acid sequences of SEQ ID NOs: 382, 383, and 384, respectively.
2. The conjugate of claim 1 , wherein the IL-21 mutein comprises the amino acid sequence of SEQ ID NO: 242.
3. The conjugate of claim 1 , wherein the IL-21 mutein comprises the amino acid sequence of SEQ ID NO: 244.
4. The conjugate of claim 1 , wherein the IL-21 mutein comprises the amino acid sequence of SEQ ID NO: 245.
5. The conjugate of claim 1 , wherein each light chain comprises a light chain variable domain (V L ) comprising the amino acid sequence of SEQ ID NO: 389 and each heavy chain comprises a heavy chain variable domain (V H ) comprising the amino acid sequence of SEQ ID NO: 388.
6. The conjugate of claim 5 , wherein each antibody light chain comprises the light chain amino acid sequence of SEQ ID NO: 391 and each antibody heavy chain comprises the heavy chain amino acid sequence of SEQ ID NO: 390.
7. The conjugate of claim 1 , wherein the IL-21 mutein is fused to the Fc of the anti-PD-1 antibody.
8. The conjugate of claim 1 , wherein the IL-21 mutein is directly fused to the Fc of the anti-PD-1 antibody without a linker.
9. The conjugate of claim 1 , wherein said conjugate comprises a single IL-21 mutein, wherein said single IL-21 mutein is fused to the C-terminus of one of the two heavy chains of the anti-PD-1 antibody.
10. The conjugate of 9 , wherein the anti-PD-1 antibody heavy chains comprise charge pair mutations.
11. The conjugate of claim 9 , wherein said single IL-21 mutein is directly fused to the C-terminus of one of the two heavy chains of the anti-PD-1 antibody without a linker.
12. The conjugate of claim 9 , wherein said single IL-21 mutein comprises the amino acid sequence of SEQ ID NO: 242.
13. The conjugate of claim 9 , wherein said single IL-21 mutein comprises the amino acid sequence of SEQ ID NO: 244.
14. The conjugate of claim 9 , wherein said single IL-21 mutein comprises the amino acid sequence of SEQ ID NO: 245.
15. The conjugate of claim 1 , wherein said conjugate
comprises a first IL-21 mutein and a second IL-21 mutein,
each fused to a C-terminus of a heavy chain of the anti-PD-1 antibody.
16. The conjugate of claim 15 , wherein said first IL-21 mutein and said second IL-21 mutein have the same amino acid sequence.
17. The conjugate of claim 15 , wherein said first IL-21 mutein and said second IL-21 mutein have different amino acid sequences.
18. The conjugate of claim 15 , wherein said first IL-21 mutein comprises the amino acid sequence of SEQ ID NO: 242.
19. The conjugate of claim 15 , wherein said first IL-21 mutein comprises the amino acid sequence of SEQ ID NO: 244.
20. The conjugate of claim 15 , wherein said first IL-21 mutein comprises the amino acid sequence of SEQ ID NO: 245.
21. A conjugate comprising:
an IL-21 mutein comprising an amino acid sequence selected from SEQ ID NOs: 233-245; and
an anti-PD-1 antibody comprising:
two light chains, each comprising an LC CDR1, LC CDR2, and LC CDR3 comprising the amino acid sequences of SEQ ID NOs: 365, 366, and 367, respectively; and
two heavy chains, each comprising an HC CDR1, HC CDR2, and HC CDR3 comprising the amino acid sequences of SEQ ID NOs: 362, 363, and 364, respectively.
22. The conjugate of claim 21 , wherein each light chain comprises a light chain variable domain (V L ) comprising the amino acid sequence of SEQ ID NO: 369 and each heavy chain comprises a heavy chain variable domain (V H ) comprising the amino acid sequence of SEQ ID NO: 368.
23. The conjugate of claim 22 , wherein each antibody light chain comprises the light chain amino acid sequence of SEQ ID NO: 371 and each antibody heavy chain amino acid sequence comprises the heavy chain of SEQ ID NO: 370.
24. A conjugate comprising an IL-21 mutein and an anti-PD-1 antibody, wherein the conjugate comprises:
(i) two antibody light chains, each comprising a light chain variable domain (V L ) comprising the amino acid sequence of SEQ ID NO: 389; and two antibody heavy chain-IL-21 mutein fusions, each comprising an amino acid sequence selected from SEQ ID NOs: 498-500 and 519; or
(ii) two antibody light chains, each comprising a V L comprising the amino acid sequence of SEQ ID NO: 389; one antibody heavy chain comprising a heavy chain amino acid sequence selected from SEQ ID NOs: 556-558, and one antibody heavy chain-IL-21 mutein fusion comprising an amino acid sequence selected from SEQ ID NOs: 501-506; or
(iii) two antibody light chains, each comprising a V L comprising the amino acid sequence of SEQ ID NO: 369; and two antibody heavy chain-IL-21 mutein fusions, each comprising an amino acid sequence selected from SEQ ID NOs: 508-512; or
(iv) two antibody light chains, each comprising a V L comprising the amino acid sequence of SEQ ID NO: 369; one antibody heavy chain comprising a heavy chain amino acid sequence selected from SEQ ID NOs: 559-561, and one antibody heavy chain-IL-21 mutein fusion comprising an amino acid sequence selected from SEQ ID NOs: 513-518.
25. The conjugate of claim 24 , comprising:
two antibody light chains, each comprising the light chain amino acid sequence of SEQ ID NO: 391;
one antibody heavy chain comprising the heavy chain amino acid sequence of SEQ ID NO: 556; and
one antibody heavy chain-IL-21 mutein fusion comprising the amino acid sequence of SEQ ID NO: 501.
26. The conjugate of claim 24 , comprising:
(i) two antibody light chains, each comprising the light chain amino acid sequence of SEQ ID NO: 391; and two antibody heavy chain-IL-21 mutein fusions, each comprising the amino acid sequence of SEQ ID NO: 499; or
(ii) two antibody light chains, each comprising the light chain amino acid sequence of SEQ ID NO: 391; and two antibody heavy chain-IL-21 mutein fusions, each comprising the amino acid sequence of SEQ ID NO: 519.
27. The conjugate of claim 24 , comprising:
two antibody light chains, each comprising the light chain amino acid sequence of SEQ ID NO: 371;
one antibody heavy chain comprising the heavy chain amino acid sequence of SEQ ID NO: 559; and
one antibody heavy chain —IL-21 mutein fusion comprising the amino acid sequence of SEQ ID NO: 513.
28. The conjugate of claim 24 , comprising:
(i) two antibody light chains, each comprising the light chain amino acid sequence of SEQ ID NO: 371; and two antibody heavy chain-IL-21 mutein fusions, each comprising the amino acid sequence of SEQ ID NO: 508; or
(ii) two antibody light chains, each comprising the light chain amino acid sequence of SEQ ID NO: 371; and two antibody heavy chain-IL-21 mutein fusions, each comprising the amino acid sequence of SEQ ID NO: 511.
29. The conjugate of claim 24 , comprising:
two antibody light chains, each comprising a V L comprising the amino acid sequence of SEQ ID NO: 389;
one antibody heavy chain comprising the heavy chain amino acid sequence of SEQ ID NO: 556; and
one antibody heavy chain-IL-21 mutein fusion comprising the amino acid sequence of SEQ ID NO: 501.
30. A pharmaceutical composition comprising:
the conjugate of any one of claims 13 , 1 , and 29 ; and
a pharmaceutically acceptable carrier, excipient, or diluent.
31. An IL-21 mutein comprising an amino acid sequence selected from SEQ ID NOs: 233-245.
32. The IL-21 mutein of claim 31 comprising the amino acid sequence of SEQ ID NO: 242.
33. The IL-21 mutein of claim 31 comprising the amino acid sequence of SEQ ID NO: 244.
34. The IL-21 mutein of claim 31 comprising the amino acid sequence of SEQ ID NO: 245.