Affinity matured CRIg variants

Patent metadata

Field	Value
Publication number	US-9234024-B2
Application number	US-38779409-A
Country	US
Kind code	B2
Filing date	May 6, 2009
Priority date	May 6, 2008
Publication date	Jan 12, 2016
Grant date	Jan 12, 2016

Abstract

The present invention concerns affinity matured CRIg variants. In particular, the invention concerns CRIg variants having increased binding affinity to C3b and retaining selective binding to C3b over C3.

First claim

What is claimed is:

1. A CRIg variant or fragment of the ECD thereof comprising an amino acid substitution at one or more amino acid positions in the amino acid sequence of SEQ ID NO:68, wherein one or more substitutions is selected from the group consisting of E8W or Y; W14F; G42D; D44H; P45F; Q601 or K; Q64R; M86Y, W, or F; Q99R or K; Q105R or K; or K110D, N or Q; and wherein the CRIg variant or fragment thereof has at least a 2-fold increased binding affinity to C3b over native sequence human CRIg of SEQ ID NO:68 or at least 2-times more potent an inhibitor of the alternative complement pathway than native sequence human CRIg of SEQ ID NO:68.

2. The variant of claim 1 which selectively binds to C3b over C3, or a fragment thereof.

3. The variant of claim 1 which has increased binding affinity to C3b over native sequence human CRIg of SEQ ID NO:68.

4. The variant of claim 3 wherein the binding affinity is increased by at least 2 fold.

5. The variant of claim 3 wherein the binding affinity is increased by at least 5 fold.

6. The variant of claim 3 wherein the binding affinity is increased by at least 10 fold.

7. The variant of claim 3 wherein the binding affinity is increased by at least 90 fold.

8. The variant of claim 1 which is a more potent inhibitor of the alternative complement pathway than native sequence human CRIg of SEQ ID NO:68.

9. The variant of claim 8 which is at least 2-times more potent than native sequence human CRIg of SEQ ID NO:68.

10. The variant of claim 8 which is at least 5-times more potent than native sequence human CRIg of SEQ ID NO:68.

11. The variant of claim 8 which is at least 10-times more potent than native sequence human CRIg of SEQ ID NO:68.

12. The variant of claim 1 comprising an amino acid substitution at one or more of amino acid positions 60, 64, 86, 99, 105 and 110 in the amino acid sequence of SEQ ID N0:68.

13. The variant of claim 1 comprising one or more substitutions selected from the group consisting of E8W, W14F, E8Y/W14F; P45F; G42D/D44H/P45F; Q60; Q64R; Q60I/Q64R; M86Y; M86W, M86F, M86W/Q99R; M86F/Q99R; K110D, K110N; Q105R/K110N; Q105R/K110Q; and Q105K/K110D.

14. The variant of claim 1 comprising one or more substitutions selected from the group consisting of Q64R/M86Y; Q60I/Q64R/E8Y; Q60I/Q64R/G42D; Q60I/Q64R/P45F; Q60I/Q64R/G42D/D44H/P45F; Q60I/Q64R/M86Y; Q60I/Q64R/Q105R; Q60I/Q64R/Q105K; Q60I/Q64R/K110N; Q60I/Q105R/K110N; M86Y/E8Y; M86Y/G42D/D44H/P45F; M86Y/P45F; M86Y/G42D/D44H/P45F; M86Y/Q99K; M86Y/Q99R; M86Y/Q105R; M86Y/Q105K; and M86Y/Q105R/K110N.

15. The variant of claim 1 comprising one or more substitutions selected from the group consisting of Q60I; Q64R; Q60I/Q64R; M86Y; Q99L; Q105K/K110D; E8W/Q105R/K110N; Q64R/M86Y; Q60I/Q64R/E8Y; Q60I/Q64R/G42D; Q60I/Q64R/P45F; Q60I/Q64R/G42D/D44H/P45F; Q60I/Q64R/M86Y; Q60I/Q64R/Q105R; Q60I/Q64R/Q105K; Q60I/Q64R/K110N; M86Y/P45F; and M86Y/Q105K.

16. The variant of claim 1 comprising a Q60I/Q64R/M86Y or Q60I/Q64R/G42D/D44H/P45F substitution.

17. A chimeric molecule comprising a variant according to claim 1 .

18. The chimeric molecule of claim 17 which is an immunoadhesin.

19. The chimeric molecule of claim 18 wherein said CRIg variant is shorter than the mature full-length CRIg of SEQ ID NO:68.

20. The chimeric molecule of claim 19 comprising a CRIg extracellular domain.

21. A pharmaceutical composition comprising a CRIg variant according to claim 1 , in admixture with a pharmaceutically acceptable excipient.

22. A pharmaceutical composition comprising an immunoadhesin according to claim 18 , in admixture with a pharmaceutically acceptable excipient.

Assignees

• Sidhu Sachdev S
• Li Bing
• Campagne Menno Van Lookeren
• Wiesmann Christian
• Genentech Inc

Inventors

• Sidhu Sachdev S
• Li Bing
• Campagne Menno Van Lookeren
• Wiesmann Christian

Classifications

• A61P37/02

  Immunomodulators · CPC title

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• A61P37/08

  Antiallergic agents (antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14) · CPC title

• A61P31/20

  for DNA viruses · CPC title

• A61P7/10

  Antioedematous agents; Diuretics · CPC title