GENE ENCODING HUMAN GLUCOKINASE MUTANT, ENZYME ENCODED BY THE SAME, RECOMBINANT VECTORS AND HOSTS, PHARMACEUTICAL COMPOSITIONS AND USES THEREOF, METHODS FOR TREATING AND PREVENTING DISEASES

• US 20110286984A1
• Filed: 07/07/2011
• Published: 11/24/2011
• Est. Priority Date: 01/07/2009
• Status: Active Grant

• Current Assignee: Yiyuan (Shenzhen) Biotech Limtied
• Original Assignee: Haidong Huang
• Inventors: Huang, Haidong
• Granted Patent: US 9,029,142 B2
• Time in Patent Office: Days
• US Class Current: 424/93.21
• CPC Class Codes: A01K 2267/0362 Animal model for lipid/glucose metabolism, e.g. obesity, type-2 diabetes
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61K 38/45 Transferases (2)
• CPC Class Codes: A61K 48/005 characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/08 for glucose homeostasis pancreatic hormones A61P5/48
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2710/10343 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 9/1205 Phosphotransferases with an alcohol group as acceptor (2.7.1), e.g. protein kinases
• CPC Class Codes: C12Y 207/01002 Glucokinase (2.7.1.2)
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

Gene encoding human glucokinase mutant is provided. The gene has the nucleotide sequence chosen from the nucleotide sequence listed as SEQ ID NO:2 and the nucleotide sequence wherein the ORF region encodes the same amino acid sequence as the one encoded by ORF region (position 487 to 1884) of SEQ ID NO:2 and the rest of the region is same as the non-ORF region of SEQ ID NO:2. Human glucokinase mutant encoded by the gene, the recombinant vectors carrying the gene, the hosts comprising the vectors, pharmaceutical compositions thereof, uses thereof, and methods for treating and preventing diseases by using the same are provided. The human glucokinase mutant encoded by the gene has higher activity than that of the wild type human glucokinase, and thus provides a new way of controlling blood glucose and/or preventing and/or treating disturbance of carbohydrate metabolism, especially preventing and treating diabetes.

Claims

1. A human glucokinase mutant-encoding gene, characterized in that the gene comprises at least one of:

a nucleotide sequence as shown in SEQ ID NO;

2; and

a nucleotide sequence that is identical to the nucleotide sequence shown in SEQ ID NO;

2 except the open reading frame (ORF) ranging from position 487 to 1884 of the nucleotide sequence shown in SEQ ID NO;

2, the ORF encoding the same amino acid sequence as that encoded by the ORF of the nucleotide sequence shown in SEQ ID NO;

2.

2. A human glucokinase mutant, characterized in that a gene encoding of the human glucokinase mutant is the human glucokinase mutant-encoding gene as set forth in claim 1.

3. A recombinant vector comprising a vector and a gene of interest carried thereon, the gene of interest comprising the human glucokinase mutant-encoding gene as set forth in claim 1.

4. The recombinant vector according to claim 3, the vector selected from a group consisting of a cloning vector, a eukaryotic expression vector, a prokaryotic expression vector, and a shuttle vector.

5. The recombinant vector according to claim 3, the vector selected from a group consisting of pIRES2-EGFP, pCMVp-NEO.BAN, pEGFT-Actin, and an adenovirus vector.

6. A host comprising a recombinant vector as set forth in claim 3.

7. The host according to claim 6, the host selected from one or more of Escherichia coli, 239 cells, min-6 cells, and human islet beta-cells.

8. A pharmaceutical composition, comprising a pharmaceutically acceptable excipient and one or more selected from:

a human glucokinase mutant-encoding gene as set forth in claim 1;

a human glucokinase mutant as set forth in claim 2;

a recombinant vector as set forth in claim 3; and

a host as set forth in claim 6.

9. The pharmaceutical composition according to claim 8, the pharmaceutical composition an injection comprising one or more selected from the human glucokinase mutant-encoding gene and the recombinant vector.

10. The pharmaceutical composition according to claim 9, the injection being a one milliliter injection comprising:

the pharmaceutically acceptable excipient comprising a phosphate buffer with a pH value of 4.0-9.0; and

one or more selected from;

10²-10¹⁰ copies of the human glucokinase mutant-encoding gene, and

10²-10¹⁰ copies of the recombinant vector.

11. The pharmaceutical composition according to claim 10, the injection comprising at least one of:

a protectant, content of the protectant 0.01-30% by weight on basis of the injection, the protectant comprising one or more selected from inositol, sorbitol, and sucrose; and

an osmotic regulator, content of the osmotic regulator allowing osmotic pressure of the injection to be 200-700 mOsm/kg, the osmotic regulator comprising at least one of sodium chloride and potassium chloride.

12. A method of at least one of:

controlling blood glucose, preventing disturbances of carbohydrate metabolism, and treating disturbances of carbohydrate metabolism, comprising;

administering an agent to a patient, the agent selected from a group consisting of;

a human glucokinase mutant-encoding gene as set forth in claim 1,

a human glucokinase mutant as set forth in claim 2,

a recombinant vector as set forth in claim 3,

a host as set forth in claim 6, and

a pharmaceutical composition as set forth in claim 8.

13. A method of at least one of:

preventing diabetes mellitus and and treating diabetes mellitus, comprising;

administering an agent to a patient, the agent selected from a group consisting of;

a human glucokinase mutant-encoding gene as set forth in claim 1,

a human glucokinase mutant as set forth in claim 2,

a recombinant vector as set forth in claim 3,

a host as set forth in claim 6, and

a pharmaceutical composition as set forth in claim 8.