Method for the identification and/or validation of receptor...

Details Method for the identification and/or validation of receptor... Method for the identification and/or validation of receptor...

C - Chemistry, Metallurgy

12

Q

C12Q 1/48 (2006.01)

Patent

CA 2544074

An in vivo method for the identification and/or validation of receptor tyrosine kinase inhibitors is described. Said method is characterised by the following steps: a) providing host cells comprising a nucleic acid construct encoding a peptide which comprises a tyrosine kinase domain of a receptor tyrosine kinase wherein said peptide lacks a transmembrane domain or a functional fragment thereof and said tyrosine kinase activity in the cytoplasma leads to proliferation arrest, b) contacting said host cells with a candidate compound and c) identification of inhibitors of said tyrosine kinase activity by cultivation of said host cells under suitable conditions such that the modulation of the tyrosine kinase activity by the candidate compound leads to cell growth.

Cette invention se rapporte à un procédé in vivo servant à l'identification et/ou à la validation d'inhibiteurs de tyrosine-kinases réceptrices. Ce procédé se caractérise par les étapes suivantes: (a) formation de cellules hôtes comprenant un produit de synthèse d'acide nucléique codant un peptide qui comprend un domaine tyrosine-kinase d'une tyrosine-kinase réceptrice, dans laquelle ce peptide est exempt d'un domaine transmembranaire ou d'un fragment fonctionnel de ce domaine et l'activité de la tyrosine-kinase dans le cytoplasme conduit à l'arrêt de la prolifération; (b) mise en contact de ces cellules hôtes avec un composé candidat; et (c) identification des inhibiteurs de l'activité de cette tyrosine-kinase par culture des cellules hôtes dans des conditions appropriées, pour que la modulation de l'activité de tyrosine-kinase par le composé candidat conduise à une croissance cellulaire.

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