Patent ID: 8349571

Claim:
A high content screening (HCS) assay for rapidly screening one or more compounds to determine functional response or pharmacological properties thereof, comprising: i) priming a cell or cell material expressing a GPCR with a sensor for a biological response; ii) contacting the compound(s)to be tested with the primed cell or cell material or contacting a cell or cell material which has been contacted with the compound(s) with the primed cell or cell material, whereby the compound(s) is to be pharmacologically analyzed as a ligand for GPCRs; iii) simultaneously or subsequently contacting with a fluorescent agonist or a fluorescent antagonist whose pharmacological properties are known, selected from a GPCR ligand moiety linked to a fluorescent moiety via a linker at a linking site which maintains ligand activity, the fluorescent moiety comprising a red absorbing BDI dye wherein the binding of the fluorescent agonist or antagonist and its associated biological response are detected or monitored in the same cell and are distinct allowing separate readout, wherein the fluorescent agonist or fluorescent antagonist is a novel fluorescent histamine receptor agonist or fluorescent histamine receptor antagonist selected from red absorbing BODIPY derivatives of GPCR ligands wherein the ligand is selected from the formula R h R hI R h2 L.h wherein R h is selected from C 1-20 hydrocarbyl including one or more heteroaryl, aryl, cycloaryl, heterocyclyl optionally together with or substituted by or including one or more heteroatoms or halo such as O, N, Cl R h1 is selected from C 0-4 alkyl R h2 is a single bond or is selected from —NR h3 —, —H 1 C(═H 2 )NH—or wherein H 1 is NR h3 or O or S and H 2 is selected from —HN, —O and —S and wherein R h3 is selected from H, C 1-3 alkyl such as CH 3 and CN and R h4 is selected from a single bond, C 1-6 ether or etheramide Or R h2 is selected from HNC(═NR h3 )NH, OCH 2 C(═O)NH, SC(═NH)NH, L.h comprises linking functionality J T which is amino, and linker linker L which is selected from C 1-12 straight chain alkyl, C 6-12 cycloalkyl or aryl and combinations thereof optionally comprising one or more heteroatoms O and optionally substituted by C 1 aliphatic.