Assay and reagents for identifying anti-proliferative agents

C - Chemistry – Metallurgy – 12 – Q

Patent

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Details

C12Q 1/68 (2006.01) C12N 1/19 (2006.01) C12N 9/16 (2006.01) C12Q 1/02 (2006.01) C12Q 1/42 (2006.01) G01N 33/50 (2006.01)

Patent

CA 2163524

The present invention makes available assays and reagents for identifying antiproliferative agents, such as mitotic and meiotic inhibitors. The present assay provides a simple and rapid screening test which relies on scoring for positive cellular proliferation as indicative of anti-mitotic or anti-meiotic activity, and comprises contacting a candidate agent with a cell which has an impaired cell-cycle checkpoint and measuring the level of proliferation in the presence and absence of the agent. The checkpoint impairment is such that it either causes premature progression of the cell through at least a portion of a cell-cycle or inhibition of normal progression of the cell through at least a portion of a cell-cycle, but can be off- set by the action of an agent which inhibits at least one regulatory protein of the cell cycle (e.g., cdc25) in a manner which counter- balances the effect of the impairment.

L'invention concerne des techniques et des réactifs utilisés pour identifier des agents antiprolifératifs tels que des inhibiteurs mitotiques et méiotiques. Ladite technique permet un criblage simple et rapide reposant sur la cotation de la prolifération cellulaire positive en tant qu'indicateur de l'activité antimitotique ou antiméiotique et consistant à mettre un agent candidat en contact avec une cellule dont le point de contrôle du cycle cellulaire est défaillant, et à mesurer le niveau de prolifération en présence de l'agent et en l'absence de celui-ci. La défaillance du point de contrôle est telle qu'elle provoque la progression prématurée de la cellule dans au moins une partie d'un cycle de cellule ou l'inhibition de la progression normale de la cellule dans au moins une partie d'un cycle de cellule, mais peut être compensée par l'action d'un agent inhibant au moins une protéine de régulation du cycle de cellule (par exemple, cdc25) de manière à contrebalancer les effets de la défaillance.

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