Functional characterization of N-octyl 4-methylamphetamine variants and related bivalent compounds at the dopamine and serotonin transporters using Ca
| Autor: | Iwona, Ruchala, Umberto M, Battisti, Vy T, Nguyen, Rita Yu-Tzu, Chen, Richard A, Glennon, Jose M, Eltit |
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| Rok vydání: | 2021 |
| Předmět: |
Serotonin Plasma Membrane Transport Proteins
Dopamine Plasma Membrane Transport Proteins Time Factors Molecular Structure Green Fluorescent Proteins Biosensing Techniques Article Methamphetamine Structure-Activity Relationship HEK293 Cells Dopamine Uptake Inhibitors Microscopy Fluorescence Humans Calcium Calcium Channels Selective Serotonin Reuptake Inhibitors |
| Zdroj: | Toxicol Appl Pharmacol |
| ISSN: | 1096-0333 |
| Popis: | The early characterization of ligands at the dopamine and serotonin transporters, DAT and SERT, respectively, is important for drug discovery, forensic sciences, and drug abuse research. 4-Methyl amphetamine (4-MA) is a good example of an abused drug whose overdose can be fatal. It is a potent substrate at DAT and SERT where its simplest secondary amine (N-methyl 4-MA) retains substrate activity at them. In contrast, N-n-butyl 4-MA is very weak, therefore it was categorized as inactive at these transporters. Here, N-octyl 4-MA and other related compounds were synthesized, and their activities were evaluated at DAT and SERT. To expedite this endeavor, cells expressing DAT or SERT were co-transfected with a voltage-gated Ca(2+) channel and, the genetically-encoded Ca(2+) sensor, GCaMP6s. Control compounds and the newly synthesized molecules were tested on these cells using an automated multi-well fluorescence plate reader; substrates and inhibitors were identified successfully at DAT and SERT. N-Octyl 4-MA and three bivalent compounds were inhibitors at these transporters. These findings were validated by measuring Ca(2+)-mobilization using quantitative fluorescence microscopy. The bivalent molecules were the most potent of the series and were further characterized in an uptake-inhibition assay. Compared to cocaine, they showed comparable potency inhibiting uptake at DAT and higher potency at SERT. These observations support a previous hypothesis that amphetamine-related (and, here, N-extended alkyl and) bivalent arylalkylamine molecules are active at monoamine transporters, showing potent activity as reuptake inhibitors, and implicate the involvement of a distant auxiliary binding feature to account for their actions at DAT and SERT. |
| Databáze: | OpenAIRE |
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