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Identification of novel inhibitors of the amino acid transporter B 0 AT1 (SLC6A19), a potential target to induce protein restriction and to treat type 2 diabetes
Author(s) -
Cheng Qi,
Shah Nishank,
Bröer Angelika,
Fairweather Stephen,
Jiang Yang,
Schmoll Dieter,
Corry Ben,
Bröer Stefan
Publication year - 2017
Publication title -
british journal of pharmacology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.432
H-Index - 211
eISSN - 1476-5381
pISSN - 0007-1188
DOI - 10.1111/bph.13711
Subject(s) - amino acid transporter , amino acid , transporter , biochemistry , chemistry , angiotensin ii receptor type 1 , biology , angiotensin ii , receptor , gene
Background 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 IC 50 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.