Compositions and methods for modulating hemostasis
View Patent ↗Novel thrombin/prothrombin protease/zymogen variants which have anticoagulation activity and methods of use thereof are disclosed.
1. A variant zymogen/protease which modulates hemostasis wherein said variant comprises amino acids 44-622 of the sequence of SEQ ID NO: 1 said variant containing at least one modification in SEQ ID NO: 1 selected from the group consisting of:
a) Ile at position 364 is replaced with Thr, or Val;
b) Val at position 365 is replaced with Ala, or Ile, or Glu and
c) Glu at position 366 is replaced with Thr, or Gly.
2. The variant zymogen/protease of claim 1 , wherein the amino acid sequence at positions 364-346 in SEQ ID NO: 1 is selected from the group consisting of Thr-Ala-Thr, Val-Ile-Glu and Ile-Glu-Gly.
3. A pharmaceutical composition comprising the variant zymogen/protease of claim 1 in a biologically compatible carrier.
4. A nucleic acid encoding the variant zymogen/protease of claim 1 .
5. A nucleic acid as claimed in claim 4 , wherein the nucleotides at positions 1121-1129 of SEQ ID NO: 2 encode amino acids selected from the group consisting of Thr-Ala-Thr, Val-Ile-Glu and Ile-Glu-Gly.
6. The variant zymogen/protease encoded by the nucleic acid of claim 5 , present in a biologically compatible carrier.
7. The nucleic acid of claim 5 cloned into an expression vector.
8. The vector of claim 7 , selected from the group consisting of an adenoviral vector, an adenovirus-associated vector, a retroviral vector, a plasmid, and a lentiviral vector.
9. A host cell expressing the vector of claim 7 .