Relaxin fusion polypeptides and uses thereof
The present invention relates to Relaxin fusion polypeptides, in particular to Relaxin 2 fusion polypeptides and uses thereof. Thus, the invention provides Relaxin fusion polypeptides, nucleic acid molecules, vectors, host cells, pharmaceutical compositions and kits comprising the same and uses of the same including methods of treatment. The polypeptides and compositions of the invention may be useful, in particular, in the treatment of cardiovascular diseases, for example for the treatment of heart failure.
1. A fusion polypeptide comprising A-L-B or B-L-A,
wherein:
A is a Relaxin A chain polypeptide or a variant thereof;
B is a Relaxin B chain polypeptide or a variant thereof; and
L is a linker polypeptide comprising at least 15 amino acids,
wherein the fusion polypeptide has Relaxin activity and further comprises a half-life extending moiety.
2. The fusion polypeptide of claim 1 , wherein the Relaxin A chain is a Relaxin 2 A chain and the Relaxin B chain is a Relaxin 2 B chain.
3. The fusion polypeptide of claim 1 , wherein the linker polypeptide L is 15 to 20 amino acids in length.
4. The fusion polypeptide of claim 1 , wherein the linker polypeptide L comprises three or more repeats of the sequence GGGGS (SEQ ID NO. 58).
5. The fusion polypeptide of claim 1 , wherein the linker polypeptide L comprises the sequence GGGGSGGGGSGGGGS (SEQ ID NO. 57).
6. The fusion polypeptide of claim 1 , wherein the linker polypeptide L consists of the sequence GGGGSGGGGSGGGGS (SEQ ID NO. 57).
7. The fusion polypeptide of claim 1 , wherein the half-life extending moiety is an immunoglobulin Fc region.
8. The fusion polypeptide of claim 7 , wherein the Fc region is derived from a human IgG1 immunoglobulin.
9. The fusion polypeptide of claim 7 , wherein the Fc region has an amino acid sequence comprising at least one of the following combinations of amino acid modifications:
(i) Fc-YTE (M252Y, S254T, T256E);
(ii) Fc-FQQ (L234F, L235Q, K322Q);
(iii) Fc-TM (L234F, L235E, P331S);
(iv) Fc-YTE-FQQ (M252Y, S254T, T256E, L234F, L235Q, K322Q);
(v) Fc-YTE-TM (M252Y, S254T, T256E, L234F, L235E, P331S),
(vi) Fc-TM-ΔTHT (L234F, L235E, P331S, D221G, K222G, T223G, H224S, T225A),
(vii) Fc-TM-ΔTHTΔK (L234F, L235E, P331S, D221G, K222G, T223G, H224S, T225A, ΔK447),
wherein the amino acid numbering is according to the EU index as in Kabat.
10. The fusion polypeptide of claim 1 , wherein the half-life extending moiety is attached to the N-terminus of the fusion polypeptide.
11. The fusion polypeptide of claim 1 , wherein the half-life extending moiety is attached to the fusion polypeptide via a connector polypeptide.
12. The fusion polypeptide of claim 11 , wherein the connector polypeptide comprises the amino acid sequence GGSP (SEQ ID NO. 56) or GGGGSGGGGSGGGGS (SEQ ID NO. 57).
13. The fusion polypeptide of claim 11 , wherein the connector polypeptide comprises the amino acid sequence GGGGSGGGGSGGGGSGGGGGS (SEQ ID NO. 69) or AAAGGGGSGGGGSGGGGSGGGGSA (SEQ ID NO. 70).
14. A pharmaceutical composition comprising the fusion polypeptide of claim 1 and a pharmaceutically acceptable excipient.
15. A fusion polypeptide having Relaxin activity, wherein the fusion polypeptide has the sequence as set forth in any one of SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, and 28.