Evidence for a Stereoselective Mechanism for Bitopic Activity by Extended-Length Antagonists of the D3 Dopamine Receptor

• Published in: ACS chemical neuroscience Vol. 11; no. 20; pp. 3309 - 3320
• Main Authors: Moritz, Amy E, Bonifazi, Alessandro, Guerrero, Adrian M, Kumar, Vivek, Free, R. Benjamin, Lane, J. Robert, Verma, Ravi Kumar, Shi, Lei, Newman, Amy Hauck, Sibley, David R
• Format: Journal Article
• Language: English
• Published: American Chemical Society 21-10-2020
• Subjects: D3 receptor; negative allosteric modulator; noncompetitive antagonist; Dopamine; stereoselectivity; bitopic
• ISSN: 1948-7193; 1948-7193
• DOI: 10.1021/acschemneuro.0c00425

The D3 dopamine receptor (D3R) has been suggested as a drug target for the treatment of a number of neuropsychiatric disorders, including substance use disorders (SUD). Many D3R-selective antagonists are bivalent in nature in that they engage two distinct sites on the receptora primary pharmacophore binds to the orthosteric site, where dopamine binds, whereas a secondary pharmacophore interacts with a unique secondary binding pocket (SBP). When engagement of the secondary pocket exerts allosteric activity, the compound is said to be bitopic. We recently reported the synthesis and characterization of two bitopic antagonists of the D3R, (±)-VK04-87 and (±)-VK05-95, which incorporated a racemic trans-cyclopropylmethyl linking chain. To gain a better understanding of the role of chirality in determining the pharmacology of such compounds, we resolved the enantiomers of (±)-VK04-87. We found that the (+)-isomer displays higher affinity for the D3R and exhibits greater selectivity versus the D2R than the (−)-isomer. Strikingly, using functional assays, we found that (+)-VK04-87 inhibits the D3R in a noncompetitive manner, while (−)-VK04-87 behaves as a purely competitive antagonist, indicating that the apparent allosteric activity of the racemate is due to the (+)-isomer. Molecular dynamic simulations of (+)-VK04-87 and (−)-VK04-87 binding to the D3R suggest that the (+)-isomer is able to interact with the SBP of the receptor whereas the (−)-isomer bends away from this pocket, thus potentially explaining their differing pharmacology. These results emphasize the importance of the linker, and its isomeric conformations, within extended-length molecules for their positioning and engagement within GPCR binding pockets.