The ADME-Tox parameters including permeability, metabolic stability, DDIs, and hepatotoxicity were carried out as described previously [9 (
link)–11 (
link), 13 (
link), 47 (
link)]. The ability of AS-1 to passively penetrate through the biological membranes was estimated by
Gentest Pre-coated PAMPA Plate System (Corning, Tewksbury, MA) and expressed as the permeability coefficient
Pe. The human metabolism of the compound AS-1 was studied using human liver microsomes (HLMs) provided by Sigma-Aldrich. The potential DDIs were predicted by luminescent
CYP3A4,
CYP2D6, and
CYP2C9 P450-Glo assays (Promega, Madison, WI). The respective strong CYP’s inhibitors, ketoconazole (KE, half maximal inhibitory concentration (IC
50) = 0.14 μM), quinidine (QD, IC
50 = 0.01 μM), and sulfaphenazole (SE, IC
50 = 0.08 μM), were used as the references. The hepatic safety of AS-1 was estimated here using hepatoma HepG2 cell growth. The cells were seeded in a 96-well plate and incubated in the presence of AS-1 at the concentration range 0.1–100 μM. One micromolar of cytostatic drug doxorubicin (DX) and 10 μM of mitochondrial toxin carbonyl cyanide 3-chlorophenylhydrazone (CCCP) were used as the references.
Kamiński K., Socała K., Zagaja M., Andres-Mach M., Abram M., Jakubiec M., Pieróg M., Nieoczym D., Rapacz A., Gawel K., Esguerra C.V., Latacz G., Lubelska A., Szulczyk B., Szewczyk A., Łuszczki J.J, & Wlaź P. (2019). N-Benzyl-(2,5-dioxopyrrolidin-1-yl)propanamide (AS-1) with Hybrid Structure as a Candidate for a Broad-Spectrum Antiepileptic Drug. Neurotherapeutics, 17(1), 309-328.
