# US 7,745,174 B2 · Hybrid hepatocyte growth factor gene having high expression efficiency of two heterotypes of hepatocyte growth factor > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 7,745,174 B2 - Document ID: US7745174B2 - Status: Granted - Kind: B2 - Application number: 12/650,860 - Filed: 2009-12-31 - Published: 2010-04-29 - Granted: 2010-06-29 - Earliest claimed: 2002-03-20 - Assignees: ViroMed Co., Ltd. - Inventors: Jong-Mook Kim; Woong Hahn; Eun-Jin Park - Web page: https://patentgrep.com/patent/US7745174B2 ## CPC - C07K 14/4753: {Hepatocyte growth factor; Scatter factor; Tumor cytotoxic factor II} - A61P 1/16: for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics - A61P 9/10: for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis - 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 ## Abstract The present invention relates to a hybrid Hepatocyte Growth Factor (HGF) gene which is prepared by inserting an inherent or foreign intron between exons 4 and 5 in HGF cDNA, which has a base sequence of SEQ ID NO: 2. The gene has high expression efficiency and simultaneously expresses two heterotypes of HGF and dHGF (deleted variant HGF). Further the gene may be used for treating or preventing ischemic or liver diseases. ## Claims ### Claim 1 A method for co-expressing two heterotypes of Hepatocyte Growth Factor (HGF) comprising transforming or transfecting to a cell a DNA construct comprising: (a) a promoter, (b) a first cDNA which has the same sequence as exons 1-4 of the human HGF gene wherein said exons 1-4 are arranged in sequential order without an intron therebetween, or degenerates thereof which do not alter the amino acid sequence encoded by said first cDNA, (c) a polynucleotide that has the same sequence as intron 4 of the HGF gene or a functional fragment thereof, and (d) a second cDNA which has the same sequence as exons 5-18 of the human HGF gene wherein said exons 5-18 are arranged in sequential order without an intron therebetween, or degenerates thereof which do not alter the amino acid sequence encoded by said second cDNA; wherein (c) is located between (b) and (d); and the HGF construct simultaneously encodes two heterotypes of human HGF. ### Claim 2 (depends on 1) The method of claim 1, wherein said intron has the same sequence as a fragment of intron 4 of the HGF gene. ### Claim 3 (depends on 2) The method of claim 2, wherein the construct comprises a nucleotide sequence not less than 90% identical to SEQ ID NO: 19. ### Claim 4 (depends on 3) The method of claim 3, wherein the construct comprises a nucleotide sequence not less than 95% identical to SEQ ID NO: 19. ### Claim 5 (depends on 4) The method of claim 4, wherein the construct comprises the sequence of SEQ ID NO: 19. ### Claim 6 (depends on 2) The method of claim 2, wherein the construct comprises a nucleotide sequence not less than 90% identical to SEQ ID NO: 20. ### Claim 7 (depends on 6) The method of claim 6, wherein the construct comprises a nucleotide sequence not less than 95% identical to SEQ ID NO: 20. ### Claim 8 (depends on 7) The method of claim 7, wherein the construct comprises the sequence of SEQ ID NO: 20. ### Claim 9 (depends on 2) The method of claim 2, wherein the construct comprises a nucleotide sequence not less than 90% identical to SEQ ID NO: 21. ### Claim 10 (depends on 9) The method of claim 9, wherein the construct comprises a nucleotide sequence not less than 95% identical to SEQ ID NO: 21. ### Claim 11 (depends on 10) The method of claim 10, wherein the construct comprises the sequence of SEQ ID NO: 21. ### Claim 12 (depends on 1) The method of claim 1, wherein the one intron has the same sequence as the full intron 4 of the HGF gene. ### Claim 13 (depends on 1) The method of claim 1, wherein the construct comprises a nucleotide sequence not less than 90% identical to SEQ ID NO: 2. ### Claim 14 (depends on 13) The method of claim 13, wherein the construct comprises a nucleotide sequence not less than 95% identical to SEQ ID NO: 2. ### Claim 15 (depends on 14) The method of claim 14, wherein the construct comprises the sequence of SEQ ID NO: 2. ### Claim 16 (depends on 1) The method of claim 1, wherein the construct further comprises a terminator sequence, a self-replication sequence, or a secretory signal. ### Claim 17 (depends on 1) The method of claim 1, wherein the expression efficiency of the construct is higher than the expression efficiency of HGF cDNA or deleted variant HGF (dHGF) cDNA. ### Claim 18 (depends on 1) The method of claim 1, wherein the expression level of the construct is about 20- to 100-fold higher than the expression level of the HGF cDNA or dHGF cDNA. ### Claim 19 (depends on 1) The method of claim 1, wherein the cell is a mammalian cell, a bacterial cell or a yeast cell. ### Claim 20 (depends on 19) The method of claim 19, wherein the cell is a mammalian cell. ### Claim 21 (depends on 20) The method of claim 20, wherein the transformation of said mammalian cell is in vivo.