Humanized monoclonal antibodies against activated protein c and uses thereof

Patent metadata

Field	Value
Publication number	US-RE49099-E
Application number	US-201316419319-A
Country	US
Kind code	E1
Filing date	Nov 27, 2013
Priority date	Nov 29, 2012
Publication date	Jun 7, 2022
Grant date	Jun 7, 2022

Abstract

Provided are humanized antibodies that selectively bind to and inhibit activated protein C without binding to or inhibiting unactivated protein C. Methods of treatment employing these antibodies are described herein.

First claim

The invention claimed is:

1. A humanized antibody binding to activated protein C comprising:
(a) a heavy light chain comprising heavy light chain CDRs represented by SEQ ID NOS: 1, 2 and 3; and wherein the light chain framework regions are represented by amino acid residues 1 to 23, 39 to 53, 61 to 92 and 102 to 112 of SEQ ID NO: 26, or having 5 or fewer conservative amino acid substitutions to SEQ ID NO: 26 framework regions, and
(b) a light heavy chain comprising light heavy chain CDRs represented by SEQ ID NOS: 4, 5 and 6 and wherein the heavy chain framework regions are represented by amino acid residues 1 to 30, 36 to 49, 69 to 100 and 108 to 118 of SEQ ID NO: 16, or having 5 or fewer conservative amino acid substitutions to SEQ ID NO: 16 framework regions.

2. The antibody of claim 1 , wherein the heavy chain framework regions are represented by SEQ ID NOS: 7, 8, 9 and 10, or having 5 or fewer conservative amino acid substitutions.

3. The antibody of claim 1 , wherein the light chain framework regions are represented by SEQ ID NOS: 11, 12, 13 and 14, or having 5 or fewer conservative amino acid substitutions.

4. The antibody of claim 2 1, wherein residue 14 49 of SEQ ID NO: 8 16 is substituted with Alanine.

5. The antibody of claim 2 1, wherein residues 11, 13 and 31 of SEQ ID NO: 9 79, 81 and 99 of SEQ ID NO: 16 are is substituted with one or more of Serine (residue 11 79), Valine (residue 13 81) and Isoleucine (residue 31 99).

6. The antibody of claim 1 , wherein said heavy chain comprises SEQ ID NOS: 16-24 NO: 16, 17, 18, 19, 20, 21 or 22.

7. The antibody of claim 3 1, wherein residue 4 of SEQ ID NO: 11 26 is substituted with Leucine.

8. The antibody of claim 3 1, wherein residue 12 72 of SEQ ID NO: 13 26 is substituted with Arginine.

9. The antibody of claim 1 , wherein said light chain comprises SEQ ID NOS: 26-30 NO: 26, 27, 28 or 29.

10. The antibody of claim 1 , wherein the antibody is a single-chain antibody or antibody fragment binding activated protein C.

11. The antibody of claim 10 , wherein the antibody fragment is further defined as Fab′, Fab, F(ab′) 2 , a single domain antibody, Fv, or scFv.

12. A cell or cell line comprising a nucleic acid encoding a humanized antibody binding to activated protein C comprising:
(a) a heavy light chain comprising heavy light chain CDRs represented by SEQ ID NOS: 1, 2 and 3; and wherein the light chain framework regions are represented by amino acid residues 1 to 23, 39 to 53, 61 to 92 and 102 to 112 of SEQ ID NO: 26, or having 5 or fewer conservative amino acid substitutions to SEQ ID NO: 26 framework regions, and
(b) a light heavy chain comprising light heavy chain CDRs represented by SEQ ID NOS: 4, 5 and 6, wherein the heavy chain framework regions are represented by amino acid residues 1 to 30, 36 to 49, 69 to 100 and 108 to 118 of SEQ ID NO: 16, or having 5 or fewer conservative amino acid substitutions to SEQ ID NO: 16 framework regions.

13. The antibody of claim 1 , dispersed in a pharmaceutically acceptable carrier.

14. A method of inhibiting activated protein C anticoagulant activity and/or amidolytic activity in a subject, comprising administering an effective amount of an antibody according to claim 1 to said subject.

15. A method of treating a subject in need of blood coagulation comprising administering an effective amount of an antibody according to claim 1 to said subject.

16. The method of claim 15 , wherein said subject is suffering from hemophilia.

17. A method of promoting hemostasis or thrombosis in a subject, comprising administrating administering an effective amount of an antibody according to claim 1 to said subject.

18. The method of claim 17 , wherein the subject is a trauma patient.

19. A kit comprising an antibody according to claim 1 .

Assignees

• Bayer Healthcare Llc

Inventors

• Zhao Xiao-Yan
• Wang Zhuozhi
• Kim Ji-Yun
• Zhu Ying
• Tebbe Jan

Classifications

• A61P7/00

  Drugs for disorders of the blood or the extracellular fluid · CPC title

• C07K16/40 Primary

  against enzymes · CPC title

• C07K2317/24

  containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered · CPC title

• G01N2333/96461

  Protein C (3.4.21.69) · CPC title

• C07K2317/567

  Framework region [FR] · CPC title