Identification and characterization of an atypical Gas-biased β2AR agonist that fails to evoke airway smooth muscle cell tachyphylaxis

G protein-coupled receptors display multifunctional signaling, offering the potential for agonist structures to promote conformational selectivity for biased outputs. For beta(2)-adrenergic receptors (beta(2)AR), unbiased agonists stabilize conformation(s) that evoke coupling to Gas (cyclic adenosine monophosphate [cAMP] production/human airway smooth muscle [HASM] cell relaxation) and ss-arrestin engagement, the latter acting to quench Gas signaling, contributing to receptor desensitization/tachyphylaxis. We screened a 40-million-compound scaffold ranking library, revealing unanticipated agonists with dihydroimidazolyl-butylcyclic urea scaffolds. The S-stereoisomer of compound C1 shows no detectable beta-arrestin engagement/signaling by four methods. However, C1-S retained Gas signaling-a divergence of the outputs favorable for treating asthma. Functional studies with two models confirmed the biasing: beta(2)AR-mediated cAMP signaling underwent desensitization to the unbiased agonist albuterol but not to C1-S, and desensitization of HASM cell relaxation was observed with albuterol but not with C1-S. These HASM results indicate biologically pertinent biasing of C1-S, in the context of the relevant physiologic response, in the human cell type of interest. Thus, C1-S was apparently strongly biased away from beta-arrestin, in contrast to albuterol and C5-S. C1-S structural modeling and simulations revealed binding differences compared with unbiased epinephrine at transmembrane (TM) segments 3,5,6,7 and ECL2. C1-S (R2 = cyclohexane) was repositioned in the pocket such that it lost a TM6 interaction and gained a TM7 interaction compared with the analogous unbiased C5-S (R2 = benzene group), which appears to contribute to C1-S biasing away from beta-arrestin. Thus, an agnostic large chemical-space library identified agonists with receptor interactions that resulted in relevant signal splitting of beta(2)AR actions favorable for treating obstructive lung disease.

Automated summary

The study identified a potentially beneficial β(2)-adrenergic receptor agonist, C1-S, for treating asthma, which diverges from the influence of β-arrestin by retaining Gas signaling, resulting in biologically relevant biasing effects.