# US 8,785,597 B2 · Mutant G-CSF fusion protein, and preparation and use thereof > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,785,597 B2 - Document ID: US8785597B2 - Status: Granted - Kind: B2 - Application number: 13/575,752 - Filed: 2010-06-30 - Published: 2013-06-27 - Granted: 2014-07-22 - Earliest claimed: 2010-06-30 - Assignees: Jiangsu T-Mab Biopharma Co., Ltd - Applicants: Xiaofang Wen; Yiliang Wu; Yefei Wang; Zhiyu Yang; Min Fan; Yujiao Wang; Xiaochun Fang; You Lu - Inventors: Xiaofang Wen; Yiliang Wu; Yefei Wang; Zhiyu Yang; Min Fan; Yujiao Wang; Xiaochun Fang; You Lu - Web page: https://patentgrep.com/patent/US8785597B2 ## CPC - C07K 14/535: Granulocyte CSF; Granulocyte-macrophage CSF - A61P 7/00: Drugs for disorders of the blood or the extracellular fluid - A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) - C07K 2319/31: fusions, other than Fc, for prolonged plasma life, e.g. albumin - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) ## Abstract The present invention relates to a mutant G-CSF fusion protein. The mutant G-CSF fusion protein is a fusion protein having the activity of stimulating the proliferation of neutrophilic granulocytes, and having a basic structure of G-CSF/carrier protein or carrier protein/G-CSF; wherein the G-CSF moiety comprises multipoint substitutions thus resulting in changes in biological activity and binding affinity. Compared with existing products, the mutant G-CSF fusion protein in the present invention has longer half-life and higher biological activity. Administration of the pharmaceutical preparation containing this mutant G-CSF fusion protein could be used in the treating neutropenia. ## Claims ### Claim 1 A mutant G-CSF fusion protein, comprising a G-CSF moiety and a carrier protein; said G-CSF moiety being derived from a human G-CSF protein with its amino acid sequence set forth in SEP ID NO: 1; said G-CSF moiety comprising at least one of the mutations of K34H, L35I, K40H and L41I; and said mutant G-CSF fusion protein stimulating proliferations of neutrophilic granulocytes. ### Claim 2 (depends on 1) The mutant G-CSF fusion protein as described in claim 1, wherein the G-CSF moiety has at least 90% sequence identity with SEQ ID NO: 1; and the carrier protein is human serum albumin, human transferrin, or antibody Fc fragment. ### Claim 3 (depends on 1) An isolated polynucleotide molecule encoding the mutant G-CSF fusion protein of claim 1. ### Claim 4 (depends on 3) An expression vector containing the isolated polynucleotide molecule of claim 3. ### Claim 5 (depends on 4) A recombinant host cell comprising the expression vector of claim 4. ### Claim 6 (depends on 5) A method of making the mutant G-CSF fusion protein comprising culturing the host cell of claim 5, and purifying the fusion protein through affinity chromatography, ion exchange chromatography and hydrophobic interaction chromatography. ### Claim 7 (depends on 1) A pharmaceutical composition comprising the mutant G-CSF fusion protein of claim 1 and at least a pharmaceutically acceptable carrier or excipient. ### Claim 8 (depends on 1) A method for treating neutropenia in mammalians, comprising administrating an effective amount of the mutant G-CSF fusion protein of claim 1. ### Claim 9 (depends on 1) The mutant G-CSF fusion protein as described in claim 1, wherein the G-CSF moiety further comprises one or more substitutions of T1A, L3T, G4Y and P5R. ### Claim 10 (depends on 9) The mutant G-CSF fusion protein as described in claim 9, wherein the G-CSF moiety has at least 90% sequence identity with SEQ ID NO: 1. ### Claim 11 (depends on 9) The mutant G-CSF fusion protein as described in claim 9, wherein the G-CSF moiety further comprises one or more substitutions of K34H, L35I, K40H, L41I, T1A, L3T, G4Y and P5R. ### Claim 12 (depends on 11) The mutant G-CSF fusion protein as described in claim 11, wherein the G-CSF moiety has at least 90% sequence identity with SEQ ID NO: 1; and the carrier protein is human serum albumin, human transferrin, or antibody Fc fragment. ### Claim 13 (depends on 11) The mutant G-CSF fusion protein as described in claim 11, wherein the G-CSF moiety comprises the amino acid sequence as set forth in SEQ ID NO: 2. ### Claim 14 (depends on 13) The mutant G-CSF fusion protein as described in claim 13, wherein the G-CSF moiety has at least 90% sequence identity with SEQ ID NO: 1; and the carrier protein is human serum albumin, human transferrin, or antibody Fc fragment. ### Claim 15 (depends on 11) The mutant G-CSF fusion protein as described in claim 11, wherein the mutant G-CSF fusion protein comprises the sequence set forth in SEQ ID NO: 4. ### Claim 16 (depends on 15) The mutant G-CSF fusion protein as described in claim 15, wherein the G-CSF moiety has at least 90% sequence identity with SEQ ID NO: 1; and the carrier protein is human serum albumin, human transferrin, or antibody Fc fragment.