Assays for modulators of asparaginyl hydroxylase

Details Assays for modulators of asparaginyl hydroxylase Assays for modulators of asparaginyl hydroxylase

C - Chemistry, Metallurgy

12

Q

C12Q 1/26 (2006.01) A61K 31/7088 (2006.01) A61K 38/44 (2006.01) A61K 45/00 (2006.01) A61K 48/00 (2006.01) C12N 9/02 (2006.01) G01N 33/68 (2006.01)

Patent

CA 2487303

A method of identifying an agent which modulates hydroxylation of hypoxia inducible factor (HIF), comprises contacting a HIF asparagine hydroxylase and a test substance in the presence of a substrate of the hydroxylase under conditions in which asparagine in the substrate is hydroxylated in the absence of the test substance; and determining hydroxylation of the substrate. Preferably the substrate is a HIF polypeptide comprising HIF-l.alpha., a fragment thereof comprising Asn 803 of HIF-l.alpha. or a peptide analogue of HIF-l.alpha. or fragment thereof comprising an asparagine equivalent to Asn 803 of HIF-l.alpha. and wherein hydroxylation of Asn 803 or of a said equivalent asparagine is determined.

L'invention concerne un procédé d'identification d'un agent qui module l'hydroxylation du facteur de transcription (HIF). Ce procédé consiste à mettre en contact une hydroxylase d'asparagine HIF et d'une substance test en présence d'un substrat de l'hydroxylase dans des conditions dans lesquelles l'asparagine du substrat est hydroxylée en l'absence de la substance test; et à déterminer l'hydroxylation du substrat. De préférence, le substrat est un polypeptide HIF comprenant HIF-l.alpha., un fragment de ce dernier contenant Asn 803 de HIF-l.alpha. ou un peptide analogue de HIF-l.alpha. ou un fragment de ce dernier comprenant un équivalent d'asparagine à Asn 803 de HIF-l.alpha., l'hydroxylation de Asn 803 ou de cet asparagine équivalent étant déterminé.

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