Extended Data Fig. 8: Verification of the activity of metabolites for two drugs, GSK1292263 and iloperidone.
From: Activity of GPCR-targeted drugs influenced by human gut microbiota metabolism
(a) Chemical transformation route for generating a GSK1292263 metabolite, GSK-459. (b) Structural elucidation of synthetic GSK-459 by NMR analyses. (c) LC-MS-based retention time comparison between synthetic standard (trace A in blue), drug metabolite from bacterial transformation (trace B in black) and co-injection (trace C in red). The EIC (with a ± 20 ppm window) for GSK-459 ion was determined using [M + H]+, m/z 459.2061. (d) MS2 similarity comparison with standard obtained from chemical reaction (top spectrum in blue) and drug metabolite from bacterial transformation (bottom spectrum in red). The predicted fragments from MS2 are indicated in panel b. (e) GPCR activity test (GPR119 agonist) on both GSK1292263 and its inactive metabolite GSK-459. n = 3 biological replicates. Data represents mean ± s.d. (f) Chemical structures of iloperidone and its metabolites. All metabolites were isolated from bacterial transformation culture broth, with their structures determined using NMR, excluding metabolite ilo-477, which was inferred by comparative tandem MS and isotope labelling studies. (g) Key 2D NMR (COSY and HMBC) correlations for the structural assignments of ilo-430. (h) GPCR activity test (DRD2 antagonist) on iloperidone and its metabolites. n = 3 biological replicates. Data represents mean ± s.d.
