The forebrain cholinergic system promotes higher brain function in part by signaling through the M1 muscarinic acetylcholine receptor (mAChR). During Alzheimer's disease (AD), these cholinergic neurons degenerate, therefore selectively activating M1 receptors could improve cognitive function in these patients while avoiding unwanted peripheral responses associated with non-selective muscarinic agonists. We describe here benzyl quinolone carboxylic acid (BQCA), a highly selective allosteric potentiator of the M1 mAChR. BQCA reduces the concentration of ACh required to activate M1 up to 129-fold with an inflection point value of 845 nM. No potentiation, agonism, or antagonism activity on other mAChRs is observed up to 100 μM. Furthermore studies in M1−/− mice demonstrates that BQCA requires M1 to promote inositol phosphate turnover in primary neurons and to increase c-fos and arc RNA expression and ERK phosphorylation in the brain. Radioligand-binding assays, molecular modeling, and site-directed mutagenesis experiments indicate that BQCA acts at an allosteric site involving residues Y179 and W400. BQCA reverses scopolamine-induced memory deficits in contextual fear conditioning, increases blood flow to the cerebral cortex, and increases wakefulness while reducing delta sleep. In contrast to M1 allosteric agonists, which do not improve memory in scopolamine-challenged mice in contextual fear conditioning, BQCA induces β-arrestin recruitment to M1, suggesting a role for this signal transduction mechanism in the cholinergic modulation of memory. In summary, BQCA exploits an allosteric potentiation mechanism to provide selectivity for the M1 receptor and represents a promising therapeutic strategy for cognitive disorders.

ABSTRACT, with a t 1/2 value of 59 min. In vitro autoradiography studies on rhesus monkey brain slices identified the highest level of binding in the cerebellum, brainstem, and meninges. Finally, as an index of central nervous system penetrability, the in vivo cerebrospinal fluid/plasma ratio was determined to be 2 to 3% in cisterna magna-ported rhesus monkeys.Migraine is one of the most prevalent and disabling neurological disorders, with characteristic symptoms that can last for several days. Despite its severity and high prevalence, migraine is not generally recognized as a serious medical condition and the societal burden is not fully appreciated. Migraine often affects people during their most productive years, which in turn burdens families and employers and ultimately affects the quality of life of the migraine sufferer. Migraine is generally agreed to be underdiagnosed and many migraineurs do not receive appropriate therapy, indicating there is significant room for improvement in the diagnosis and management of migraine.An overall improvement in migraine treatment occurred Article, publication date, and citation information can be found at

A growing body of evidence has implicated the calcitonin gene-related peptide (CGRP) receptors in migraine pathophysiology. With the approval of monoclonal antibodies targeting CGRP or the CGRP receptor, the inhibition of CGRP-mediated signaling emerged as a promising approach for preventive treatments of migraine in adults. Recently, small-molecule anti-CGRP treatments have shown efficacy for treating migraine. The current studies aimed to characterize the pharmacologic properties of ubrogepant, an orally bioavailable CGRP receptor antagonist for the acute treatment of migraine. In a series of ligand-binding assays, ubrogepant exhibited a high binding affinity for native (K i 5 0.067 nM) and cloned human (K i 5 0.070 nM) and rhesus CGRP receptors (K i 5 0.079 nM), with relatively lower affinities for CGRP receptors from rat, mouse, rabbit, and dog. In functional assays, ubrogepant potently blocked human a-CGRP2stimulated cAMP response (IC 50 of 0.08 nM) and exhibited highly selective antagonist activity for the CGRP receptor compared with other members of the human calcitonin receptor family. Furthermore, the in vivo CGRP receptor antagonist activity of ubrogepant was evaluated in a pharmacodynamic model of capsaicin-induced dermal vasodilation (CIDV) in rhesus monkeys and humans. Results demonstrated that ubrogepant produced concentration-dependent inhibition of CIDV with a mean EC 50 of 3.2 and 2.6 nM in rhesus monkeys and humans, respectively. Brain penetration studies with ubrogepant in monkeys showed a cerebrospinal fluid:plasma ratio of 0.03 and low CGRP receptor occupancy. In summary, ubrogepant is a competitive antagonist with high affinity, potency, and selectivity for the human CGRP receptor. SIGNIFICANCE STATEMENT Ubrogepant is a potent, selective, orally delivered, smallmolecule competitive antagonist of the human CGRP. In vivo studies using a pharmacodynamic model of CIDV in rhesus monkeys and humans demonstrated that ubrogepant produced concentration-dependent inhibition of CIDV, indicating a predictable pharmacokinetic-pharmacodynamic relationship.