Functional bioassay for g-protein coupled receptor agonists...

C - Chemistry – Metallurgy – 12 – N

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C12N 5/10 (2006.01) C12Q 1/02 (2006.01) G01N 33/50 (2006.01) G01N 33/566 (2006.01)

Patent

CA 2220715

Simple, rapid high-throughput functional bioassays for identifying agents that act as either agonists or antagonists of G-protein coupled receptors (GPCRs) are disclosed. In the methods of the invention, a test composition is contacted with an indicator cell expressing a GPCR and at least one parameter of cellular metabolism of the indicator cells is measured to identify a test compound(s) in the test composition as a receptor agonist or antagonist. The assays can be used to screen libraries of test compounds to identify therapeutically useful agonists or antagonists of GPCRs involved in disease conditions. The assays can also be used to identify ligands of "orphan" GPCRs whose natural ligands are unknown. Methods of generating indicator cells expressing GPCRs, and isolated populations of such indicator cells, are also disclosed.

Dosages biologiques fonctionnels simples, rapides et à rendement élevé, destinés à l'identification d'agents servant d'agonistes ou d'antagonistes des récepteurs couplés de la protéine G (GPCR). Selon les procédés de l'invention, on met une composition d'essai au contact d'une cellule indicatrice exprimant un GPCR, et on mesure au moins un paramètre du métabolisme cellulaire des cellules indicatrices afin de déterminer si un ou plusieurs composés d'essai présents dans la composition d'essai sont des agonistes ou antagonistes desdits récepteurs. Ces dosages permettent de rechercher, dans des banques de composés d'essai, des agonistes ou antagonistes intéressants sur le plan thérapeutique des GPCR impliqués dans différents états pathologiques. Lesdits dosages permettent également d'identifier des ligands de GPCR "orphelins" dont on ignore les ligands naturels. On a également prévu des procédés de production de cellules indicatrices exprimant les GPCR, ainsi que des populations isolées de cellules indicatrices de ce type.

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