Optimized monoclonal antibodies against tissue factor pathway inhibitor (TFPI)

Patent metadata

Field	Value
Publication number	US-RE47150-E
Application number	US-201115208498-A
Country	US
Kind code	E1
Filing date	Mar 1, 2011
Priority date	Mar 1, 2010
Publication date	Dec 4, 2018
Grant date	Dec 4, 2018

Abstract

Isolated monoclonal antibodies that bind human tissue factor pathway inhibitor (TFPI) are provided. Isolated nucleic acid molecules encoding monoclonal antibodies that bind TFPI are also contemplated. Pharmaceutical compositions comprising the anti-TFPI monoclonal antibodies and methods of treating deficiencies or defects in coagulation by administration of the antibodies are also provided. Methods of producing the antibodies are also provided.

First claim

We claim:

1. An isolated human monoclonal IgG antibody that binds specifically to human tissue factor pathway (TFPI) inhibitor, wherein the antibody comprises at least one or more amino acid substitution substitutions, comprising:
(a) a human heavy chain comprising:
a CDR 1 region with the amino acid sequence shown in SEQ ID NO: 41, optionally comprising a substitution at N32 relative to SEQ ID NO: 3;
a CDR2 region with the amino acid sequence shown in SEQ ID NO: 42, optionally comprising a substitution selected from the group consisting of K55 and D62 relative to SEQ ID NO: 3;
a CDR3 region that binds specifically to TFPI and has the amino acid sequence shown in SEQ ID NO: 43; optionally comprising a substitution at Y112 relative to SEQ ID NO: 3,
optionally, an amino acid substitution at G44 relative to SEQ ID NO: 3; and

(b) a human light chain comprising:
a CDR1 region with the amino acid sequence shown in SEQ ID NO: 44, optionally comprising a substitution of a position selected from the group consisting of S32 and N35 and combinations thereof relative to SEQ ID NO: 4;
a CDR2 region with the amino acid sequence shown in SEQ ID NO: 45;
a CDR3 region that binds specifically to TFPI and has the amino acid sequence shown in SEQ ID NO: 46;

wherein the one or more amino acid substitutions is selected from the group consisting of:
N32 relative to SEQ ID NO: 3 within the CDR1 region of the human heavy chain;
K55 relative to SEQ ID NO: 3 within the CDR2 region of the human heavy chain;
D62 relative to SEQ ID NO: 3 within the CDR2 region of the human heavy chain;
Y112 relative to SEQ ID NO: 3 within the CDR3 region of the human heavy chain;
G44 relative to SEQ ID NO: 3 within the human heavy chain;
S32 relative to SEQ ID NO: 4 within CDR1 of the human light chain; and
N35 relative to SEQ ID NO: 4 within CDR1 of the human light chain.

2. An isolated human monoclonal IgG antibody that binds specifically to human tissue factor pathway inhibitor (TFPI), wherein the antibody comprises:
(a) a human heavy chain comprising:
a CDR 1 region with the amino acid sequence shown in SEQ ID NO: 41, comprising a substitution of N32 relative to SEQ ID NO: 3,
a CDR2 region with the amino acid sequence shown in SEQ ID NO: 42, comprising at least one substitution relative to SEQ ID NO: 3 selected from the group consisting of K55Y and D62R and combinations thereof;
a CDR3 region that binds specifically to TFPI and has the amino acid sequence shown in SEQ ID NO: 43 comprising a substitution relative to SEQ ID NO: 43 of Y112D relative to SEQ ID NO: 3, and
a substitution relative to SEQ ID No. 3 of G44S; and

(b) a human light chain comprising:
a CDR1 region with the amino acid sequence shown in SEQ ID NO: 44, comprising at least one substitution selected from the group consisting of S32R and N35I relative to SEQ ID NO: 4 and combinations thereof;
a CDR2 region with the amino acid sequence shown in SEQ ID NO: 45,
a CDR3 region that binds specifically to TFPI and has the amino acid sequence shown in SEQ ID NO: 46.

3. The antibody of claim 1 , wherein the antibody comprises the amino acid substitution of N32 relative to SEQ ID NO: 3 within the CDR1 region of the human heavy chain, and wherein the substitution at N32 is N32D.

4. The antibody of claim 1 , wherein the antibody comprises the amino acid substitution of K55 relative to SEQ ID NO: 3 within the CDR2 region of the heavy chain, and wherein the substitution of K55 is K55Y.

5. The antibody of claim 1 , wherein the antibody comprises the amino acid substitution of G44 within the human heavy chain, and wherein the substitution of G44 is G44S.

6. The antibody of claim 1 , wherein the antibody comprises the amino acid substitution of D62 relative to SEQ ID NO: 3 within the CDR2 region of the human heavy chain, and wherein the substitution of D62 is D62R.

7. The antibody of claim 1 , wherein the antibody comprises the amino acid substitution of Y112 relative to SEQ ID NO: 3 within the CDR3 region of the human heavy chain, and wherein the substitution of Y112 is Y112D.

8. The antibody of claim 1 , wherein the antibody comprises the amino acid substitution of S32 relative to SEQ ID NO:4 within the CDR1 region of the human light chain, and wherein the substitution of S32 is S32R.

9. The antibody of claim 1 , wherein the antibody comprises the amino acid substitution of N35 relative to SEQ ID NO: 4 within the CDR1 region of the human light chain, and wherein the substitution of N35 is N35I.

10. The antibody of claim 1 , wherein the amino acid at position 1 of the heavy chain relative to SEQ ID NO: 3 is Q.

11. The antibody of claim 1, wherein the antibody is an IgG antibody.

12. A method of treating a subject suffering from hemophilia A or hemophilia B, comprising administering to the subject a therapeutically effective amount of an isolated human monoclonal antibody that binds specifically to human tissue factor pathway (TFPI) inhibitor, wherein the antibody comprises one or more amino acid substitutions, comprising:
(a) a human heavy chain comprising:
a CDR 1 region with the amino acid sequence shown in SEQ ID NO: 41;
a CDR2 region with the amino acid sequence shown in SEQ ID NO: 42;
a CDR3 region that binds specifically to TFPI and has the amino acid sequence shown in SEQ ID NO: 43; and
(b) a human light chain comprising:
a CDR 1 region with the amino acid sequence shown in SEQ ID NO: 44;
a CDR2 region with the amino acid sequence shown in SEQ ID NO: 45;
a CDR3 region that binds specifically to TFPI and has the amino acid sequence shown in SEQ ID NO: 46;
wherein the one or more amino acid substitutions is selected from the group consisting of:
N32 relative to SEQ ID NO: 3 within the CDR1 region of the human heavy chain;
K55 relative to SEQ ID NO: 3 within the CDR2 region of the human heavy chain;
D62 relative to SEQ ID NO: 3 within the CDR2 region of the human heavy chain;
Y112 relative to SEQ ID NO: 3 within the CDR3 region of the human heavy chain:
G44 relative to SEQ ID NO: 3 within the human heavy chain;
S32 relative to SEQ ID NO: 4 within CDR1 of the human light chain; and
N35 relative to SEQ ID NO: 4 within CDR1 of the human light chain.

13. A method of treating a subject suffering from hemophilia A or hemophilia B, comprising administering to the subject a therapeutically effective amount of an isolated human monoclonal antibody that binds specifically to human tissue factor pathway inhibitor (TFPI), wherein the antibody comprises:
(a) a human heavy chain comprising:
a CDR1 region with the amino acid sequence shown in SEQ ID NO: 41, comprising a substitution of N32 relative to SEQ ID NO: 3;
a CDR2 region with the amino acid sequence shown in SEQ ID NO: 42, comprising at least one substitution selected from the group consisting of K55Y, D62R, and a combination thereof, relative to SEQ ID NO: 3;
a CDR3 region that binds specifically to TFPI and has the amino acid sequence shown in SEQ ID NO: 43 comprising a substitution of Y112D relative to SEQ ID NO: 3; and
a substitution relative to SEQ ID NO: 3 of G44S; and
(b) a human light chain comprising:
a CDR1 region with the amino acid sequence shown in SEQ ID NO: 44, comprising at least one substitution selected from the group consisting of S32R, N35I, and a combination thereof, relative to SEQ ID NO: 4;
a CDR2 region with the amino acid sequence shown in SEQ ID NO: 45; and
a CDR3 region that binds specifically to TFPI and has the amino acid sequence shown in SEQ ID NO: 46.

14. The method of claim 12, wherein the subject is suffering from hemophilia A.

15. The method of claim

Assignees

• Bayer Healthcare Llc

Inventors

• Wang Zhuozhi
• Pan Junliang
• Grudzinska-Goebel Joanna
• Votsmeier Christian
• Tebbe Jan
• Birkenfeld Joerg
• Wobst Nina
• Brückner Simone
• Steinig Susanne
• Scholz Peter

Classifications

• A61P7/02

  Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors · CPC title

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P7/04

  Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents · CPC title

• C07K2317/92

  Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value · CPC title

• C07K2317/55

  Fab or Fab' · CPC title