RT Journal Article SR Electronic T1 Identification of Positive Allosteric Modulators of the D1 Dopamine Receptor That Act at Diverse Binding Sites JF Molecular Pharmacology JO Mol Pharmacol FD American Society for Pharmacology and Experimental Therapeutics SP 1197 OP 1209 DO 10.1124/mol.118.113175 VO 94 IS 4 A1 Kathryn D. Luderman A1 Jennie L. Conroy A1 R. Benjamin Free A1 Noel Southall A1 Marc Ferrer A1 Marta Sanchez-Soto A1 Amy E. Moritz A1 Blair K. A. Willette A1 Tim J. Fyfe A1 Prashi Jain A1 Steve Titus A1 Lisa A. Hazelwood A1 Jeffrey Aubé A1 J. Robert Lane A1 Kevin J. Frankowski A1 David R. Sibley YR 2018 UL http://molpharm.aspetjournals.org/content/94/4/1197.abstract AB The D1 dopamine receptor is linked to a variety of neuropsychiatric disorders and represents an attractive drug target for the enhancement of cognition in schizophrenia, Alzheimer disease, and other disorders. Positive allosteric modulators (PAMs), with their potential for greater selectivity and larger therapeutic windows, may represent a viable drug development strategy, as orthosteric D1 receptor agonists possess known clinical liabilities. We discovered two structurally distinct D1 receptor PAMs, MLS6585 and MLS1082, via a high-throughput screen of the NIH Molecular Libraries program small-molecule library. Both compounds potentiate dopamine-stimulated G protein- and β-arrestin-mediated signaling and increase the affinity of dopamine for the D1 receptor with low micromolar potencies. Neither compound displayed any intrinsic agonist activity. Both compounds were also found to potentiate the efficacy of partial agonists. We tested maximally effective concentrations of each PAM in combination to determine if the compounds might act at separate or similar sites. In combination, MLS1082 + MLS6585 produced an additive potentiation of dopamine potency beyond that caused by either PAM alone for both β-arrestin recruitment and cAMP accumulation, suggesting diverse sites of action. In addition, MLS6585, but not MLS1082, had additive activity with the previously described D1 receptor PAM “Compound B,” suggesting that MLS1082 and Compound B may share a common binding site. A point mutation (R130Q) in the D1 receptor was found to abrogate MLS1082 activity without affecting that of MLS6585, suggesting this residue may be involved in the binding/activity of MLS1082 but not that of MLS6585. Together, MLS1082 and MLS6585 may serve as important tool compounds for the characterization of diverse allosteric sites on the D1 receptor as well as the development of optimized lead compounds for therapeutic use.