Non-conserved residues dictate dopamine transporter selectivity for the potent synthetic cathinone and psychostimulant MDPV. (1st December 2021)

• Record Type: Journal Article
• Authors: Steele, Tyler W.E.; Spires, Zachary; Jones, Charles B.; Glennon, Richard A.; Dukat, Małgorzata; Eltit, Jose M.
• Is Part Of: Neuropharmacology. Volume 200(2021)
• Publication Date: 2021-12-01
• Subjects: hDAT -- dDAT -- Drugs of abuse -- Structure -- Monoamine transporters -- Calcium channels -- Flakka -- Bath salts -- 3D molecular modeling -- α-PVP -- MDPPP -- Synthetic cathinones; Neuropsychopharmacology -- Periodicals; Autonomic Agents -- Periodicals; Neuropsychopharmacologie -- Périodiques; Neuropsychopharmacology; Periodicals; Electronic journals; 615.78
• DOI: 10.1016/j.neuropharm.2021.108820
• Languages: English
• ISSNs: 0028-3908

Abstract

Abstract: Clandestine chemists are currently exploiting the pyrrolidinophenone scaffold to develop new designer drugs that carry the risk of abuse and overdose. These drugs promote addiction through the rewarding effects of increased dopaminergic neurotransmission. 3, 4-Methylenedioxypyrovalerone (MDPV) and its analogs are illicit psychostimulants of this class that are ∼50-fold more potent than cocaine at inhibiting the human dopamine transporter (hDAT). In contrast, MDPV is a weak inhibitor at both the human serotonin transporter (hSERT) and, as it is shown here, the Drosophila melanogaster DAT (dDAT). We studied three conserved residues between hSERT and dDAT that are unique in hDAT (A117, F318, and P323 in dDAT), and one residue that is different in all three transporters (D121 in dDAT). hDAT residues were replaced in the dDAT sequence at these positions using site-directed mutagenesis and stable cell lines were generated expressing these mutant transporters. The potencies of MDPV and two of its analogs were determined using a Ca 2+ -mobilization assay. In this assay, voltage-gated Ca 2+ channels are expressed to sense the membrane electrical depolarization evoked when dopamine is transported through DAT. Each individual mutant slightly improved MDPV's potency, but the combination of all four increased its potency ∼100-fold (2 log units) in inhibiting dDAT activity. Molecular modeling and docking studies were conducted to explore the possible mode of interaction between MDPV and DAT in silico. Two of the studied residues (F318 and P323) are at the entrance of the S1 binding site, whereas the other two (A117 and D121) face the aryl moiety of MDPV when bound to this site. Therefore, these four non-conserved residues can influence MDPV selectivity not only by stabilizing binding, but also by controlling access to its binding site at DAT. Highlights: Analogs of MDPV are frequent-appearing designer drugs. Four non-conserved residues of hDAT account for MDPV high potency. These four residues are marginally implicated in dopamine recognition. Access and interaction with the S1 site might control MDPV selectivity at DAT.