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Novel biological substance nesfatin and its related substances and uses thereof

A technology of substances and related factors, applied in the direction of DNA/RNA fragments, cells modified by introducing foreign genetic materials, genetic material components, etc., can solve physiological and pharmacological effects without reporting, without showing NEF feeding regulation and/or body weight Reconciliation related reporting and other issues

Inactive Publication Date: 2008-11-05
TEIJIN PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the NEFA polypeptide has a signal sequence at its amino terminal side, showing the possibility of extracellular secretion (医学の已み Takahashi Kazuo, Fujikatsu 2005, Volume 213, No. 6, pp. 549-554), but there is no report on the physiological and pharmacological effects produced by secretion out of cells
Also, there are no reports showing that NEF is involved in regulation of food intake and / or body weight

Method used

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  • Novel biological substance nesfatin and its related substances and uses thereof
  • Novel biological substance nesfatin and its related substances and uses thereof
  • Novel biological substance nesfatin and its related substances and uses thereof

Examples

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preparation example Construction

[0240] In the preparation method of the above-mentioned polypeptide of the present invention, in order to be suitable for expression, transcription, translation, etc. in transformants or transgenic non-human animals, the above-mentioned DNA sequence, plasmid, virus, etc. can be modified or changed more. For example, based on the synonymity of the genetic code (code), the entire protein coding region can be substituted with nucleotides. Such a sequence can be estimated from the amino acid sequence of a NESFATIN polypeptide or the like or the base sequence of a gene encoding the polypeptide, and assembled according to the following conventional synthesis methods. The above synthetic method can be carried out substantially according to the method of Itakura et al. (Itakura et al., Science 198, 1056, 1977) and the method of Crea et al. (Crea et al., Proc. Natl. Acad. Sci. USA75, 5765, 1978). Therefore, the gene encoding the above-mentioned NESFATIN polypeptide and the like used in...

Embodiment 1

[0320] Example 1 Cloning of genes induced by PPARγ agonist stimulation

[0321] In order to identify genes whose expression is regulated by PPARγ and which act on feeding and / or body weight regulation, genes that are specifically induced when stimulated with PPARγ agonists in SQ-5 cells (mainly from human lung cancer cells) were cloned and extracted from Among them, genes having a possibility of functioning as secreted factors were screened.

[0322] The method for obtaining genes whose expression is specifically induced by PPARγ in non-small cell lung cancer can be carried out according to the method of Satoh et al., Oncogene (England), 2002, volume 21, pages 2171-2180. This method is briefly described below. SQ-5 (Riken Biological Resource Center (Riken Bioliso-Scentent) RBC0110) is in use for 10 -4 Troglitazone (Sankyo Co., Ltd.), one of the M PPARγ agonists, was stimulated to culture for 48 hours, and Poly(A)+RNA ( mRNA), double-stranded cDNA was prepared by reverse tra...

Embodiment 2

[0329] Example 2 Stimulation with PPARγ agonist to induce expression of NEFA gene in cultured cell lines

[0330] In order to confirm the induction of the NEFA gene by troglitazone, the expression analysis of the NEFA gene was performed by Northern blotting using the PPARγ-expressing cell lines HTB185 and SQ5 and the adipocyte line 3T3-L1.

[0331] SQ-5 (Riken Bioresource Center RBC0110) was obtained by subculture with RPMI1640 medium (Inbitologien-BIL, product number 11875-085) containing 10% fetal bovine serum (Inbitologien-GIBGO). Human brain medulloblastoma cell line HTB185 (D283Med: ATCC HTB185) and mouse embryonic fibroblast cell line (preadipocyte cell line) 3T3-L1 cell line (ATCC CL-173) induced by insulin, dexamethasone, and IBMX The DMEM medium containing 10% fetal bovine serum (Invitrogen-GIBCO, product number 11995-040) was used for subculture. 10 suspended in 10ml medium 6 Each cell was seeded in a 10 cm culture dish, the cells were allowed to attach to the matr...

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Abstract

The present invention relates to a novel method of obtaining a factor involved in appetite control and / or body weight control, as well as genes obtained by said method, polypeptides encoded by said genes, or novel polypeptides obtained from the information on polypeptides encoded by said genes as a means for treating, controlling or diagnosing diseases associated with eating disorders and / or bodyweight control. Also the present invention relates to substances that inhibit the effects of said genes or said polypeptides as a means for treating, controlling or diagnosing diseases associated with appetite control and / or body weight control. By using thiazolidine diones having a PPAR agonist activity, genes and polypeptides involved in appetite regulation and / or body weight reduction can be obtained. NESFATIN or the like obtained by said method can be used as a means for treating, controlling or diagnosing diseases associated with eating disorders and / or body weight control.

Description

technical field [0001] The present invention relates to novel methods for obtaining factors associated with regulation of food intake and / or body weight regulation. The present invention also relates to the polypeptides related to food intake regulation and / or body weight regulation obtained by the method, nucleic acid molecules encoding the polypeptides, and the treatment, prevention, diagnosis method. The present invention also relates to methods for treating, preventing, and diagnosing diseases associated with increased appetite and / or increased weight gain using a substance that inhibits the action of the polypeptide or gene. The present invention further relates to an animal model of diseases related to feeding regulation and / or body weight regulation obtained by inhibiting the polypeptide, the gene or their activity, and a compound for regulating the action or expression of the polypeptide using the animal model screening method. The present invention also relates to ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/00A01K67/027A61K38/22A61K39/395A61K45/00A61K48/00A61P3/04C07K14/47C12N1/15C12N1/19C12N1/21C12N5/10C12N15/09C12P21/02C12Q1/02C12Q1/68G01N33/15G01N33/50
CPCA01K2217/05A61K38/16A61K48/00C07K14/47C07K16/18C12N15/113C12N2310/11C12Q1/6883G01N33/6893G01N2500/00G01N2800/02G01N2800/044G01N2800/303A61P1/16A61P11/00A61P15/12A61P19/06A61P3/10A61P35/00A61P3/04A61P3/06A61P41/00A61P9/10A61P9/12C12N15/00A61K38/22
Inventor 森昌朋清水弘行江口广志山本真则
Owner TEIJIN PHARMA CO LTD
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