Tissue factor pathway inhibitor antibodies and uses thereof
The invention relates to antibodies, and antigen-binding fragments thereof, that specifically bind TFPI and inhibit an activity thereof. Such antibodies and fragments are useful for treating bleeding disorders and shortening clotting time.
1. An isolated antibody or antigen-binding fragment thereof that specifically binds to an epitope in Kunitz Domain 2 (K2) of Tissue Pathway Factor Inhibitor (TFPI), wherein the antibody is selected from the group consisting of an antibody comprising:
(a) a heavy chain variable region (VH) comprising a VH complementarity determining region one (CDR-H1) comprising the amino acid sequence of SEQ ID NO:38, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:39, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO:40, and a light chain variable region (VL) comprising a VL complementarity determining region one (CDR-L1) comprising the amino acid sequence of SEQ ID NO:33, a CDR-L2 comprising the amino acid sequence of SEQ ID NO:34, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO:35;
(b) a VH comprising the amino acid sequence of SEQ ID NO:63 and a VL comprising the amino acid sequence of SEQ ID NO:36;
(c) a heavy chain consisting of the amino acid sequence of SEQ ID NO:64 and a light chain consisting of the amino acid sequence of SEQ ID NO:37;
(d) a VH comprising the amino acid sequence of SEQ ID NO:41 and a VL comprising the amino acid sequence of SEQ ID NO: 36; and
(e) a heavy chain consisting of the amino acid sequence of SEQ ID NO: 42 and a light chain consisting of the amino acid sequence of SEQ ID NO: 37.
2. The antibody, or antigen-binding fragment thereof, of claim 1 , wherein said antibody, or antigen-binding fragment thereof, binds to TFPI with a binding affinity (K D ) value of from about 1×10 −8 M to about 1×10 −10 M.
3. The antibody or antigen-binding fragment thereof of claim 1 , wherein said antibody or antigen-binding fragment thereof: (i) decreases clotting time as measured in a plasma based dilute prothrombin time assay; (ii) reduces clotting time in whole blood as measured by thromboelastrography; (iii) increases thrombin generation; (iv) increases FXa activity in the presence of TFPI; (v) enhances platelet accumulation in the presence of TFPI; (vi) increases fibrin generation in the presence of TFPI; or (vii) any combination thereof.
4. The antibody or antigen-binding fragment thereof of claim 3 , wherein said plasma or whole blood is deficient in Factor VIII or Factor IX.
5. A pharmaceutical composition comprising an antibody or antigen-binding fragment thereof of claim 1 , and a pharmaceutically acceptable carrier or excipient.
6. The antibody, or antigen-binding fragment thereof, of claim 1 , wherein said VH is encoded by the nucleic acid sequence of the insert present in the plasmid deposited under ATCC Accession No. PTA-122329, and said VL is encoded by the nucleic acid sequence of the insert present in the plasmid deposited under ATCC Accession No. PTA-122328.
7. The antibody, or antigen-binding fragment thereof, of claim 1 , wherein said epitope comprises residues Ile105, Arg107, Leu131, Cys106, Gly108, Cys130, Gly132, Asp102, Arg112, Tyr127, Gly129, Met134, and Glu138, according to the numbering of SEQ ID NO: 2 .
8. An isolated antibody or antigen-binding fragment thereof that specifically binds the K2 of TFPI, wherein said antibody or antigen-binding fragment comprises the CDR-H1, CDR-H2, and CDR-H3 sequences of SEQ ID NO: 63, and the CDR-L1, CDR-L2, and CDR-L3 sequences of SEQ ID NO: 36.
9. An isolated antibody or antigen-binding fragment thereof that specifically binds the K2 of TFPI, wherein said antibody or antigen-binding fragment consists of a heavy chain consisting of the amino acid sequence of SEQ ID NO:64 and a light chain consisting of the amino acid sequence of SEQ ID NO:37.