Prokineticin polypeptides, related compositions and methods

a technology of prokineticin and polypeptide, applied in the field of prokineticin polypeptide, related compositions and methods, can solve the problems of reducing the efficacy of these regulators as therapeutics, limiting the utility of these regulators for therapeutic use, etc., and achieve the effect of inhibiting promoting the production of prokineticin receptor signals

Inactive Publication Date: 2005-04-07
RGT UNIV OF CALIFORNIA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Methods of recombinantly preparing these snake and frog polypeptides, or of recombinantly preparing other polypeptides containing 10 cysteines, have not previously been described, limiting the utility of these regulators for therapeutic use.
Additionally, snake and frog polypeptides could elicit antibodies if administered to mammals that would likely reduce their efficacy as therapeutics.

Method used

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  • Prokineticin polypeptides, related compositions and methods
  • Prokineticin polypeptides, related compositions and methods
  • Prokineticin polypeptides, related compositions and methods

Examples

Experimental program
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example i

Identification, Preparation and Characterization of Prokineticins 1 and 2

[0130] This example shows the cloning, recombinant expression, purification and biological activities of human prokineticins 1 and 2, as well as modifications thereof.

[0131] Materials and Methods

[0132] RNA Blot

[0133] Human multiple tissue RNA blots containing normalized samples of polyA RNA were used as described by the manufacture's instructions (Clontech). The blots were probed with random primer-labeled probes (nucleotides 1-550 and 1-1178 for prokineticin 1 and prokineticin 2 cDNAs), and signals were visualized by exposing to Kodak XAR film.

[0134] Production, Refolding and Purification of Recombinant Prokineticins

[0135] The coding sequences for mature prokineticins were cloned into prokacyotic expression vector pGEX-3X (Pharmacia). The extra nucleotides between the factor Xa protease digestion site of GST (Glutathione-s-transferase) tag and mature prokineticins were removed by site-directed mutagenesi...

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Abstract

The invention provides isolated polypeptides that stimulate gastroeintestinal smooth muscle contraction, including human prokineticin 1 and human prokineticin 2 polypeptides, and functional fragments and modifications thereof. Also provided are methods of stimulating gastrointestinal smooth muscle contraction in a mammal, by administering to the mammal an effective amount of a prokineticin polypeptide. The invention also provides nucleic acid molecules encoding a prokineticin polypeptide, and antibodies that selectively bind a prokineticin polypeptide. Further provided are methods of identifying a prokineticin receptor ligand, agonist or antagonist.

Description

BACKGROUND OF THE INVENTION [0001] The main function of gastrointestinal (GI) smooth muscle is to mix and propel intralumenal contents, which enables efficient digestion of food, progressive absorption of nutrients, and eventual evacuation of residual components. The activity of GI smooth muscle is regulated by intrinsic and extrinsic neural signals, including classical neurotransmitters, co-existing neuropeptides, and circulating peptide hormones. In addition, a number of humoral agents including histamine, serotonin, and adenosine that are produced by nonneural GI cells also influence the activity of smooth muscle cells. [0002] A number of clinical conditions are associated with altered GI motility, including irritable bowel syndrome, diabetic gastroparesis, postoperational ileus, chronic constipation, gastrointestinal reflux disease, chronic diarrhea, infectious diseases, malabsorptive disorders, inflammatory bowel disorders, and intestinal cancers. The identification of regulato...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K38/00A61K39/00A61P1/00G01N33/50A61P29/00A61P31/00A61P35/00C07K14/47C07K16/18C12N1/15C12N1/19C12N1/21C12N5/10C12N15/09C12N15/12C12P21/02C12Q1/02G01N33/15G01N33/53G01N33/566
CPCA61K38/00C07K14/47A61K39/00A61P1/00A61P29/00A61P31/00A61P35/00
Inventor ZHOU, QUN-YONGEHLERT, FREDERICK
Owner RGT UNIV OF CALIFORNIA
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