US 2008/0305500 A1 · NOVEL CELL-BASED ASSAYS FOR IDENTIFYING ENHANCERS OR INHIBITORS OF T1R TASTE RECEPTORS (T1R2/T1R3 SWEET) AND UMAMI (T1R1/T1R3 UMAMI) TASTE RECEPTORS

Abstract

This invention relates to improved assays for identifying modulators (enhancers or inhibitors) of sweet (T1R2/T1R3) and umami (T1R1/T1R3) taste receptors. These receptors may comprise the endogenous T1Rs e.g., from humans or rodents, or may comprise functional variants such as chimeric taste receptors comprising the extracellular portion of one T1R or a variant or fragment thereof, either T1R1 or T1R2, and the transmembrane portion of another T1R or a variant or fragment thereof, either T1R1 or T1R2, preferably associated with a T1R3 polypeptide and a suitable G protein. The subject assays preferably use endogenous taste or gastrointestinal cells which express T1R taste receptors or recombinant cell which express such T1Rs, for example mammalian cells or Xenopus oocytes. The modulators (enhancers or inhibitors) identified according to the subject assays are useful as taste modulators (which is confirmed in taste tests) and/or as therapeutics for treating conditions such as diabetes, obesity, weight control, fat metabolism, glucose metabolism, insulin metabolism, satiety and/or the release of satiety peptides such as GLP-1.

Claims53

Dependent claims

An assay method for identifying a modulator (enhancer or inhibitor) of an activator the sweet (T1R2/T1R3) or umami (T1R1/T1R3) taste receptor comprising

(i) preincubating a sweet receptor or umami receptor respectively comprised of T1R2 and T1R3 polypeptides or T1R1 and T1R3 polypeptides with at least one first compound that is being screened as a potential modulator of said sweet or umami receptor;
(ii) after said preincubation step contacting said preincubated sweet or umami receptor with a suboptimal concentration of at least one second compound that activates said sweet or umami receptor, wherein a suboptimal concentration of said second compound is a concentration below the saturation concentration that results in maximal detectable activity of the sweet or umami receptor;
(iii) detecting sweet or umami receptor activity after step (ii) and comparing said activity to sweet or umami receptor activity that is detected when said sweet or umami receptor is contacted with said suboptimal concentration of said at least one second compound that activates the sweet or umami receptor in the absence of said preincubation step with said at least one first compound;
(iv) identifying the screened first compound as a putative enhancer or inhibitor of said second compound on the activation of the sweet or umami receptor if the detected sweet or umami receptor activity is respectively greater or lesser in the presence of the preincubation step (ii) than in the absence of said preincubation step (ii).

1 more element

Dependencies are inferred from the claim text.

Novel cell-based assays for identifying enhancers or inhibitors of t1r taste receptors (t1r2/t1r3 sweet) and umami (t1r1/t1r3 umami) taste receptors

Life of this patent

Novel cell-based assays for identifying enhancers or inhibitors of t1r taste receptors (t1r2/t1r3 sweet) and umami (t1r1/t1r3 umami) taste receptors

Life of this patent

Abstract

Claims53