Genetically modified TFPI and method of treating coagulation using the same
AssigneeFudan University
InventorsDuan Ma, Jingui Mu, Jiping Wang, Huijun Wang, Wang Liang
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Abstract
The present invention provides long half life genetically modified TFPI sequences (LTFPI) for anticoagulation. On the genetically modified TFPI sequence, the lysine at the carboxy-terminal sites 241, 254, 260 and 261 are replaced by alanin and the amino acid asparagine at glycosylation sites 117, 167, 228 and the amino acids serine and threonine at glycosylation sites 174 and 175 are substitutionally mutated. The present invention also provides methods of making the LTFPI through high efficient LTFPI expression from yeast production system.
Claims4
A synthesized polypeptide optimized for a Pichia pastoris expression system, wherein said polypeptide is capable of anticoagulation, comprising a synthesized Long-half-life Tissue Factor Pathway Inhibitor (LTFPI) polypeptide represented by SEQ ID NO:
- 2 which is mutated from human wild-type Tissue Factor Pathway Inhibitor (TFPI), wherein the lysines of the polypeptide at the carboxy-terminal residues 241, 254, 260 and 261, the asparagines at glycosylation residues 117, 167, 228, and the serine and threonine at glycosylation residues 174 and 175 are all substitutionally mutated to alanine, and wherein the polypeptide is optimized for Pichia pastoris expression.
A pharmaceutical composition for anticoagulation comprising a pharmaceutically effective amount of a genetically modified Long-half-life Tissue Factor Pathway Inhibitor (LTFPI) polypeptide represented by SEQ ID NO:
- 2 which is mutated from human wild type Tissue Factor Pathway Inhibitor (TFPI) wherein the lysines of the polypeptide at the carboxy-terminal residue 241, 254, 260 and 261, and amino acid asparagine at glycosylation residues 117, 167, 228 and amino acids serine and threonine at glycosylation residues 174 and 175 are all substitutionally mutated by amino acid alanine and wherein the polypeptide is optimized for Pichia pastoris expression.
A method for treating coagulation comprising administering to a host a pharmaceutically effective amount of a genetically modified Long-half-life Tissue Factor Pathway Inhibitor (LTFPI) polypeptide represented by SEQ ID NO:
- 2, wherein the lysines of the polypeptide at the carboxy-terminal residues 241, 254, 260 and 261, the asparagines at glycosylation residues 117, 167, 228, and the serine and threonine at glycosylation residues 174 and 175 are all substitutionally mutated to alanine, and wherein the polypeptide is optimized for Pichia pastoris expression.
Dependencies are inferred from the claim text.