Exploring the molecular determinants for subtype-selectivity of 2-amino-1,4,5,6-tetrahydropyrimidine-5-carboxylic acid analogs as betaine/GABA transporter 1 (BGT1) substrate-inhibitors
| Autor: | Kickinger, Stefanie, Al-Khawaja, Anas, Haugaard, Anne Stæhr, Lie, Maria E. K., Bavo, Francesco, Löffler, Rebekka, Damgaard, Maria, Ecker, Gerhard F., Frølund, Bente, Wellendorph, Petrine |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
GABA Plasma Membrane Transport Proteins
Proton Magnetic Resonance Spectroscopy DIVERSITY lcsh:Medicine Molecular Dynamics Simulation Crystallography X-Ray Article HIGH-AFFINITY Structure-Activity Relationship Computational Chemistry TARGETS CEREBRAL-CORTEX Humans lcsh:Science Transporters in the nervous system IDENTIFICATION GABA TRANSPORTER lcsh:R LOCALIZATION Stereoisomerism BINDING-SITE Pharmacodynamics Drug Design lcsh:Q PHARMACOLOGICAL CHARACTERIZATION ANTICONVULSANT ACTION |
| Zdroj: | Scientific Reports, Vol 10, Iss 1, Pp 1-15 (2020) Scientific Reports Kickinger, S, Al-Khawaja, A, Haugaard, A S, Lie, M E K, Bavo, F, Loeffler, R, Damgaard, M, Ecker, G F, Frolund, B & Wellendorph, P 2020, ' Exploring the molecular determinants for subtype-selectivity of 2-amino-1,4,5,6-tetrahydropyrimidine-5-carboxylic acid analogs as betaine/GABA transporter 1 (BGT1) substrate-inhibitors ', Scientific Reports, vol. 10, no. 1, 12992 . https://doi.org/10.1038/s41598-020-69908-w |
| DOI: | 10.1038/s41598-020-69908-w |
| Popis: | We have previously identified 2-amino-1,4,5,6-tetrahydropyrimidine-5-carboxylic acid (ATPCA) as the most potent substrate-inhibitor of the betaine/GABA transporter 1 (BGT1) (IC50 2.5 mu M) reported to date. Herein, we characterize the binding mode of 20 novel analogs and propose the molecular determinants driving BGT1-selectivity. A series of N-1-, exocyclic-N-, and C-4-substituted analogs was synthesized and pharmacologically characterized in radioligand-based uptake assays at the four human GABA transporters (hGATs) recombinantly expressed in mammalian cells. Overall, the analogs retained subtype-selectivity for hBGT1, though with lower inhibitory activities (mid to high micromolar IC50 values) compared to ATPCA. Further characterization of five of these BGT1-active analogs in a fluorescence-based FMP assay revealed that the compounds are substrates for hBGT1, suggesting they interact with the orthosteric site of the transporter. In silico-guided mutagenesis experiments showed that the non-conserved residues Q299 and E52 in hBGT1 as well as the conformational flexibility of the compounds potentially contribute to the subtype-selectivity of ATPCA and its analogs. Overall, this study provides new insights into the molecular interactions governing the subtype-selectivity of BGT1 substrate-inhibitors. The findings may guide the rational design of BGT1-selective pharmacological tool compounds for future drug discovery. |
| Databáze: | OpenAIRE |
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