β2AR-IN-15 is a selective β2-adrenergic receptor ( β2AR ) antagonist with a K d of 1.7 μM. β2AR-IN-15 binds to an intracellular β2AR region overlapping the G-protein binding site, enhancing orthosteric inverse agonist binding while negatively modulating binding of orthosteric agonists with EC 50 values of 1.9 and 0.48 μM. β2AR-IN-15 shows inhibitory effect on cAMP production and β-arrestin recruitment to activated β2AR. β2AR-IN-15 can be used for the research of cardiovascular and pulmonary diseases [1].
In Vitro, β2AR-IN-15 (Compound 15) acts as a negative allosteric modulator for human β2AR reconstituted in nanodiscs, decreasing orthosteric agonist binding with an EC50 of 1.9 μM and increasing inverse agonist binding with an EC50 of 0.48 μM[1]. β2AR-IN-15 binds directly to purified human β2AR with a Kd of 1.7 μM and a 1:1 binding stoichiometry[1]. β2AR-IN-15 competes with the intracellular-binding nanobody Nb60 for binding to human β2AR, indicating its binding site is located on the receptor's intracellular region[1]. β2AR-IN-15 inhibits transducer-promoted high-affinity agonist binding to human β2AR reconstituted in nanodiscs, with EC50 values of 2.8 μM for Gαβγ and 2.2 μM for β-arrestin1, showing no bias between the two transducers[1]. β2AR-IN-15 (1.25-50 μM; 20 min) inhibits isoproterenol-stimulated cAMP production in HEK-293 cells with endogenous β2AR and β-arrestin recruitment to chimeric β2V2R, causing 7.1-fold and 7.4-fold right shifts of the isoproterenol EC50, respectively[1]. β2AR-IN-15 (1.25-50 μM; 20 min) selectively inhibits agonist-stimulated β-arrestin recruitment to human β2AR, with significantly reduced or no activity on β1AR, V2R, VIPR, and AT1R[1].
References:
[1]. Ahn S, et al. Allosteric "beta-blocker" isolated from a DNA-encoded small molecule library. Proc Natl Acad Sci U S A. 2017;114(7):1708-1713.
In Vitro, β2AR-IN-15 (Compound 15) acts as a negative allosteric modulator for human β2AR reconstituted in nanodiscs, decreasing orthosteric agonist binding with an EC50 of 1.9 μM and increasing inverse agonist binding with an EC50 of 0.48 μM[1]. β2AR-IN-15 binds directly to purified human β2AR with a Kd of 1.7 μM and a 1:1 binding stoichiometry[1]. β2AR-IN-15 competes with the intracellular-binding nanobody Nb60 for binding to human β2AR, indicating its binding site is located on the receptor's intracellular region[1]. β2AR-IN-15 inhibits transducer-promoted high-affinity agonist binding to human β2AR reconstituted in nanodiscs, with EC50 values of 2.8 μM for Gαβγ and 2.2 μM for β-arrestin1, showing no bias between the two transducers[1]. β2AR-IN-15 (1.25-50 μM; 20 min) inhibits isoproterenol-stimulated cAMP production in HEK-293 cells with endogenous β2AR and β-arrestin recruitment to chimeric β2V2R, causing 7.1-fold and 7.4-fold right shifts of the isoproterenol EC50, respectively[1]. β2AR-IN-15 (1.25-50 μM; 20 min) selectively inhibits agonist-stimulated β-arrestin recruitment to human β2AR, with significantly reduced or no activity on β1AR, V2R, VIPR, and AT1R[1].
References:
[1]. Ahn S, et al. Allosteric "beta-blocker" isolated from a DNA-encoded small molecule library. Proc Natl Acad Sci U S A. 2017;114(7):1708-1713.