Ethylmethylhydroxypyridine Succinate Is an Inhibitor but Not a Substrate of ABCB1 and SLCO1B1.

2-Ethyl-6-methyl-3-hydroxypyridine succinate (EMHPS, Mexidol) is an original antioxidant and an anti-ischemic drug with the possibility of wide applications in the complex therapy of diseases, accompanied by the development of oxidative stress and ischemia; for example, ischemic stroke, chronic cerebral ischemia, and chronic heart failure. The use of EMHPS in the complex therapy of the above diseases may cause the development of drug-drug interactions, particularly pharmacokinetic interactions at the level of transporter proteins. In the present study, we evaluated the interaction of EMHPS with ABCB1 and SLCO1B1. In Caco-2 cells, it was shown that EMHPS is not a substrate of ABCB1 and that it does not affect its expression, but at the same time, it inhibits the activity of this transporter. Its inhibitory activity was inferior to verapamil-a classic inhibitor of ABCB1. In HEK293 and HEK293-SLCO1B1 cells, it was shown that EMHPS is not a substrate of SLCO1B1 either, but that it inhibited the activity of the transporter. However, its inhibitory activity was inferior to the classic inhibitor of SLCO1B1-rifampicin. Furthermore, it was found out that EMHPS does not affect SLCO1B1 expression in HepG2 cells. The approach proposed by the FDA (2020) and the International Transporter Consortium (2010) was used to assess the clinical significance of the study results. The effect of EMHPS on SLCO1B1 and the systemic inhibition of ABCB1 by EMPHS are not clinically significant, but ABCB1 inhibition by EMHPS in the gastrointestinal tract should be tested in vivo through clinical trials.

Mechanisms of Regulation of the P-Glycoprotein Transporter Protein Functioning under the Action of Nitric Oxide.

Mechanisms of regulation of the P-glycoprotein (Pgp) transporter under the action of nitric oxide (NO) were studied in Caco-2 cells. S-Nitrosoglutathione (GSNO) was used as a NO donor, which was added to the cells at concentrations 1, 10, 50, 100, and 500 µM and incubated for 3, 24, or 72 h. The amount of Pgp was analyzed using Western blotting, activity was determined by monitoring transport of its substrate, fexofenadine. The study showed that a short-term exposure to GSNO for 3 h at 500 µM concentration caused increase in the concentration of peroxynitrite in Caco-2 cells, which reduced the activity, but not the amount of Pgp. Increase in the duration of exposure to 24 h increased the amount and activity of Pgp at GSNO concentrations of 10 and 50 µM, increased the amount without increasing activity at 100 µM concentration, and decreased the amount of the transporter protein at 500 µM. Duration of exposure to GSNO of 72 h at concentration of 10 µM resulted in the increase of the amount and activity of Pgp, while at concentration of 100 and 500 µM it decreased the amount of the transport protein. At the same time, it was shown using specific inhibitors that the increase in the amount of Pgp under the influence of low concentrations of GSNO was realized through the NO-cGMP signaling pathway, and the effect of the higher concentration of GSNO and the respective development of nitrosative stress was realized through Nrf2 and the constitutive androstane receptor.