Factor VII polypeptides that are modified and uses thereof
Nucleic acids encoding modified factor VII polypeptides, vectors and cells containing the nucleic acids, uses of the nucleic acids, methods of making the encoded polypeptides, and methods of treatment are provided. The encoded modified FVII polypeptides include Factor VIIa and other forms of Factor VII. Among the encoded modified FVII polypeptides provided are those that have altered activities, typically altered procoagulant activity, including increased procoagulant activities.
1. A method of catalyzing activation of a Factor X, comprising contacting the Factor X with a modified factor VII (FVII) polypeptide, wherein the modified FVII polypeptide has 1-7 amino acid replacements compared to an unmodified FVII polypeptide of SEQ ID NO: 3, wherein the modified FVII polypeptide comprises a replacement at a position corresponding to position Q286 in a FVII polypeptide having the sequence of amino acids set forth in SEQ ID NO: 3, wherein the modification at position Q286 is an amino acid replacement with Arg (R)(Q286R), wherein the modified FVII polypeptide has at least 90% sequence identity to a polypeptide of any of SEQ ID NOS.: 1-3, wherein the modified FVII polypeptide has a kinetic constant for catalytic activity greater than that of the unmodified FVII polypeptide of SEQ ID NO: 3, and wherein the modified FVII polypeptide is a single chain zymogen, a zymogen-like two-chain polypeptide, or a fully activated two-chain form.
2. The method of claim 1 , wherein the modified FVII polypeptide has only 1, 2, 3, 4, 5, 6 or 7 amino acid modifications compared to an unmodified FVII polypeptide of SEQ ID NO: 3.
3. The method of claim 1 , wherein the modified FVII polypeptide comprises a M298Q amino acid replacement.
4. The method of claim 1 , wherein the contacting is in vitro.
5. The method of claim 1 , wherein the contacting is in vivo.
6. The method of claim 1 , wherein the modified FVII polypeptide comprises amino acid replacements selected from among Q286R, Q286R/H257A, Q286R/S222A, Q286R/S222A/H257A, Q286R/S222A/Gla Swap FIX, Q286R/M298Q, Q286R/H257A/Gla Swap FIX, Q286R/S222A/H257A/Gla Swap FIX, Q286R/Q366V, Q286R/A292N/A294S/Q366V, A175S/Q286R/Q366V, S222A/Q286R/Q366V, H257S/Q286R, H257S/Q286R/Q366V, S222A/H257A/Q286R/Q366V, Q286R/H373A, S222A/H257A/Q286R/M158Q, Q286R/K341D, Q286R/Q366D, Q286R/Q366N, Q286R/H373F, T128N/P129A/Q286R/H373F, S52A/S60A/Q286R, S52A/S60A/Q286R/H373F, T239V/Q286R, T239V/Q286R/H373F, T239I/Q286R/H373F, T128N/P129A/A175S/Q286R, T128N/P129A/Q286R/M298Q, and A122N/G124S/A175S/Q286R.