Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Synthesis and characterisation of a new benzamide-containing nitrobenzoxadiazole as a GSTP1-1 inhibitor endowed with high stability to metabolic hydrolysis

doi: 10.1080/14756366.2019.1617287

Figure Lengend Snippet: Assessment of metabolic stability of compounds 3 and 4 in human liver microsomes (HLMs). Compound 3 (panel A) or 4 (panel B) was incubated with pooled, mixed-gender HLMs (0.05 mg of protein/mL) in 0.1 M potassium phosphate buffer, pH 7.4 (-•-) or buffer only (-□-) at 37 °C for various time intervals ( n = 5). The final concentration of each NBD derivative in the incubation mixture was 10 µM. To evaluate the role of CES on depletion of 3 in human liver microsomal incubations, the NBD derivative (10 µM) was incubated at 37 °C with pooled, mixed-gender HLMs (0.05 mg of protein/mL) in 0.1 M potassium phosphate buffer (pH 7.4) for 0 and 10 min, both in the absence and in the presence of 50 µM benzil (panel C). The data are expressed as percentage of compound remaining at each time compared with time 0 min, and represent the mean ± SD of three independent determinations. Error bars smaller than the symbols are not visible. *** P < 0.001 vs. control (minus benzil) at 10 min.

Article Snippet: The role of human liver microsomal CES in hydrolysis of compound 3 was investigated by incubating the test compound (10 μM) at 37 °C for 0 and 10 min in 0.2 mL of 0.1 M potassium phosphate buffer (pH 7.4) containing 0.05 mg of protein/mL of pooled, mixed-gender HLMs (Xenotech LLC) and the nonselective CES inhibitor benzil (50 μM) ; the incubation protocol and sample preparation for HPLC analysis were the same as described above.

Techniques: Incubation, Concentration Assay, Control