Cheng, QiShah, NishankBröer, AngelikaFairweather, StephenJiang, YangSchmoll, DieterCorry, Ben AlexanderBröer, Stefan2017-04-060007-1188http://hdl.handle.net/1885/114511BACKGROUND AND PURPOSE The neutral amino acid transporter B(0) AT1 (SLC6A19) has recently been identified as a possible target to treat type 2 diabetes and related disorders. B(0) AT1 mediates the Na(+) -dependent uptake of all neutral amino acids. For surface expression and catalytic activity, B(0) AT1 requires coexpression of collectrin (TMEM27). In this study, we established tools to identify and evaluate novel inhibitors of B(0) AT1. EXPERIMENTAL APPROACH A CHO-based cell line was generated, stably expressing collectrin and B(0) AT1. Using this cell line, a high-throughput screening assay was developed, which uses a fluorescent dye to detect depolarisation of the cell membrane during amino acid uptake via B(0) AT1. In parallel to these functional assays, we ran a computational compound screen using AutoDock4 and a homology model of B(0) AT1 based on the high-resolution structure of the highly homologous Drosophila dopamine transporter. KEY RESULTS We characterized a series of novel inhibitors of the B(0) AT1 transporter. Benztropine was identified as a competitive inhibitor of the transporter showing an IC50 of 44 ± 9 μM. The compound was selective with regard to related transporters and blocked neutral amino acid uptake in inverted sections of mouse intestine. CONCLUSION AND IMPLICATIONS The tools established in this study can be widely used to identify new transport inhibitors. Using these tools, we were able to identify compounds that can be used to study epithelial transport, to induce protein restriction, or be developed further through medicinal chemistry.The work reported here was supported by a sponsored research contract with Sanofi (Germany).application/pdf© 2017 The British Pharmacological Society2017-0310.1111/bph.13711