CPC G01N 33/502 (2013.01) [G01N 2333/726 (2013.01)] | 27 Claims |
1. A high-throughput assay system for identifying one or more compounds from a group of compounds wherein the one or more compounds positively modulates the activity of an olfactory receptor induced by an agonist odorant compound, comprising:
a) one or more isolated cells, each cell expressing one mammalian olfactory receptor, wherein the one or more cells comprises a target olfactory receptor that is activated by one or more odorant,
b) a first compound that binds to the olfactory receptor and that activates the olfactory receptor, and
c) at least one second compound that binds to the receptor non-competitively relative to the first compound and that enhances the activity of the receptor exposed to the first compound when compared to the activity of the receptor exposed to the first compound in the absence of the second compound,
wherein the olfactory receptor comprises a polypeptide that
comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 2, 4, 6, or 8; or
is encoded by a nucleic acid molecule comprising a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 1, 3, 5, or 7, or the reverse complement thereof,
wherein the first compound is an agonist odorant compound of the olfactory receptor,
wherein the first compound is a musk or a floral muguet compound,
wherein the first compound and the second compound are different compounds,
wherein the second compound is not an agonist of the receptor,
wherein the second compound has the structure:
in the form of any one of its stereoisomers or a mixture thereof,
wherein R represents a C3-11 linear alkyl group optionally substituted by a C1-4 alkyl carboxylester group or a hydroxyl group, a C4-11 branched alkyl group optionally substituted by a C1-3 alkoxy group, a C6-12 linear or branched alkenyl or alkadienyl group, a phenyl group substituted by one or two C1-3 alkyl groups, a C5-8 alicyclic alkenyl group or a benzyloxymethyl group, or
wherein the second compound is at least one positive allosteric modulator selected from the group consisting of (E)-dec-2-enal, 2-phenylpropanal; (E)-but-2-enal; 3-methylbenzaldehyde;
3-(1,3-benzodioxol-5-yl)-2-methylpropanal; (+−)-3-(4-methyl-3-penten-1-yl)-3-cyclohexene-1-carbaldehyde, (+−)-4-(4-methyl-3-penten-1-yl)-3-cyclohexene-1-carbaldehyde, and mixtures thereof; heptanal, 4-Propan-2-ylbenzaldehyde; (3R)-3,7-dimethyloct-6-enal; (+)-(3S)-3-[(1R)-4-methyl-3-cyclohexen-1-yl]butanal, (+)-(3R)-3-[(1R)-4-methyl-3-cyclohexen-1-yl]butanal, and mixtures thereof; hexanal; 2,6-dimethylhept-5-enal; benzaldehyde; 2-methyl-3-(4-methylphenyl)propanal; 3,5,6-trimethyl-3-cyclohexene-1-carbaldehyde, 2,4,6-trimethyl-3-cyclohexene-1-carbaldehyde, and mixtures thereof, 4-ethylbenzaldehyde; 6-methoxy-2,6-dimethylheptanal; (E)-non-2-enal; and combinations thereof,
wherein the second compound is a positive allosteric modulating compound, and
wherein the activity of the receptor is synergistically enhanced by the combination of the first compound and the second compound.
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