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Screening method

A screening method and amino acid technology, applied in the field of peptides, can solve the problems of unknown relaxin-3, weight control without relaxin-3, unknown function, etc.

Inactive Publication Date: 2007-02-21
EISIA R&D MANAGEMENT CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

On the other hand, GPR100 has been reported as a systemically expressed receptor (Non-Patent Documents 8 and 9); however, its function is still unknown
[0007] On the other hand, relaxin-3 has been reported to be present in a region of the brain called the pons (Non-Patent Document 6), and it has been considered that relaxin-3 may function as an intracerebral peptide in the central nervous system; however , there is no report on whether relaxin-3 controls food intake or whether relaxin-3 is involved in weight control
Additionally, it is unknown whether relaxin-3 is involved in obesity

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0335] Example 1: Preparation of polynucleotides encoding SALPR

[0336] Based on the nucleic acid sequence represented by SEQ ID NO: 3, isolation of the polynucleotide encoding SALPR was performed as follows. In SEQ ID NO: 3, 1857 base pairs are shown and the region known to encode SALPR is from position 361 to position 1770 (1410 base pairs, 470 amino acid residues) (GenBank accession number: NM_016568). In order to isolate genes by polymerase chain reaction (PCR), PCR primers represented by SEQ ID NO: 5 and SEQ ID NO: 6 were prepared according to ordinary methods.

[0337] Using human genomic DNA (Roche Diagnostics) as a template, using a set of PCR primers represented by SEQ ID NO: 5 and SEQ ID NO: 6, using (Expand High Fidelity PCR System (Roche Diagnostics) according to the manufacturer's instructions, 30 repeated cycles ( 98° C. for 1 minute, 57° C. for 1 minute, and 72° C. for 3 minutes) for PCR reaction. As a result, a DNA fragment of about 1400 base pairs was obtain...

Embodiment 2

[0339] Embodiment 2: the preparation of retroviral vector plasmid

[0340] pBabe Puro (Morgenstern, J.P. and Land, H. Nucleic Acids Res. Vol. 18, 3587-3596 (1990) (SEQ ID NO: 7) was digested with SalI and ClaI to remove the SV40 promoter-puro (r) region, The fragment that obtains fills in with Klenow fragment.IRES-hyg(r) region is inserted at the cleavage site, thereby obtains pBabeXIH, wherein IRES-hyg(r) region is to adopt NsiI and XbaI enzyme digestion to excise from pIREShyg (Clontech) and use Filled with T4 polymerase.

[0341] pBabeXIH was digested with SspI and BamHI to remove the 5'-LTR-packaging signal. The 5'LTR-CMV promoter-packaging signal was inserted at the cleavage site to obtain pBabeCLXIH, wherein the 5'LTR-CMV promoter-packaging signal was excised from pCLXSN (IMGENEX) using SspI and BamHI digestion.

Embodiment 3

[0342] Example 3: Preparation of retroviral vector plasmids for SALPR gene transfer

[0343] The retroviral expression plasmid pBabeCLXIH described in Example 2 above was digested with the restriction enzyme HpaI. A polynucleotide encoding SALPR was inserted at the cleavage site to obtain pBabeCL(SALPR) IH, wherein the polynucleotide was excised from the pCR2.1-SALPR obtained in the above Example 1 by EcoRV digestion and filled in with T4 polymerase acquired( figure 1 ).

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Abstract

The present inventors have found that relaxin-3 has a feeding-stimulating activity, a body weight increasing activity, and fat weight increasing activity by intracerebroventricularly administering relaxin-3 to rats and observing the amount of feeding, body weight and fat weight, and so on after the administration. According to the present invention, a polypeptide having useful effects in stimulating feeding, increasing body weight, and fattening; a therapeutic agent containing said polypeptide; a method of screening for a compound, a substance, or a salt thereof which activates or suppresses a receptor of said polypeptide; a kit for said screening; and an agent which comprises a substance which inhibits expression of said polypeptide, such as a feeding-surpressing agent, a therapeutic agent for the treatment of obesity, and a therapeutic agent for the treatment of diabetes.

Description

field of invention [0001] The present invention relates to polypeptides having beneficial effects in stimulating feeding (feeding), increasing body weight and fattening; therapeutic agents containing said polypeptides; methods of screening compounds, substances or salts thereof which activate or inhibit said polypeptide receptors; A kit for said screening; and a medicament comprising a substance that inhibits the expression of said polypeptide, such as a feeding inhibitor, a therapeutic agent for treating obesity, and a therapeutic agent for treating diabetes. Background technique [0002] Ingestion (eating) is an essential behavior for animals to survive. Obesity is thought to be the result of the failure to control or balance food intake and energy expenditure in our current socially fed age. Obesity is of increasing social concern as it is a risk factor for lifestyle diseases and various other diseases. Although there are basic treatments such as diet therapy and exerci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09C12Q1/02C07K14/435C07K14/705A61K38/00A61P1/00A61P3/04G01N33/15
Inventor 飞弹隆之关矢智子泽井彻生木尚志高桥英机小笹美智子原田耕吉新井彻
Owner EISIA R&D MANAGEMENT CO LTD
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