Pharmaceutical composition containing, as active ingredient, granulocyte-colony stimulating factor mutant protein or transferrin fusion protein thereof
The present invention relates to a fusion protein in which transferrin is peptide-bonded to a terminal of a granulocyte-colony stimulating factor (G-CSF) protein or a G-CSF mutant protein in which the 116 th threonine is substituted with cysteine in the amino acid sequence of the G-CSF. Specifically, the granulocyte-colony stimulating factor (G-CSF) mutant protein of the present invention or the transferrin fusion protein thereof displays a significantly increased specific activity and blood stability, compared with the conventional human G-CSF, and has a higher purification efficiency than the conventional PEGylated G-CSF characterized by the extended half-life, so that it can be advantageously used for preventing or treating ischemic diseases or neutropenia.
1. An expression vector comprising a gene encoding a fusion protein in which transferrin is peptide-bonded to a G-CSF (granulocyte-colony stimulating factor) mutant protein having at least 80% amino acid sequence homology to SEQ ID NO: 1 and in which the 116th threonine is substituted with a cysteine,
wherein the expression vector has a sequence selected from the group consisting of SEQ ID NOS: 6, 7, 8 and 21.
2. The expression vector of claim 1 , wherein the transferrin of the fusion protein encoded by the gene has a deletion of a signal peptide at the amino terminal region and/or does not have a native signal peptide at the amino terminal region.
3. The expression vector of claim 1 , wherein the gene encoding the fusion protein has a substitution of a native nucleotide sequence for a restriction enzyme recognition site with a replacement nucleotide sequence encoding the same amino acid as the one encoded by the native nucleotide sequence.
4. The expression vector of claim 1 , wherein the gene encoding the protein additionally contains a restriction enzyme recognition site at the end of the gene.
5. The expression vector of claim 1 , wherein the expression vector has a sequence according to SEQ ID NO: 6.
6. The expression vector of claim 1 , wherein the expression vector has a sequence according to SEQ ID NO: 7.
7. The expression vector of claim 1 , wherein the expression vector has a sequence according to SEQ ID NO: 8.
8. The expression vector of claim 1 , wherein the expression vector has a sequence according to SEQ ID NO: 21.
9. A method for treating or preventing ischemic disease or neutropenia, the method comprising the step of administering the expression vector of claim 1 to a subject in need thereof.
10. The method of claim 9 , wherein the subject has cancer, the method comprising administering the expression vector to the subject to treat or prevent neutropenia associated with the cancer or treatment thereof.
11. The method of claim 9 , wherein the subject has ischemic disease or is at risk of developing ischemic disease, the method comprising administering the expression vector to the subject to treat or prevent the ischemic disease.