Stereoselectivity of evodiamine enantiomers in neuroprotective activity, pharmacokinetics and the ability across the blood-brain barrier.

Abstract

Evodiamine, a chiral quinazoline alkaloid in the traditional Chinese medicine Evodiae fructus, exhibited efficacy for CNS diseases. In this study, the pure enantiomers of evodiamine were prepared in large quantities via chemical resolution. Their structures were elucidated by MS, NMR and ECD. The optical purity was determined to be as high as 99.8 %. The differences of the enantiomers in protective effect against neuronal cell injury were evaluated using MTT assay. Notably, R-(-)-evodiamine showed better neuroprotection effects against H₂O₂-induced damage in PC12 cells. An efficient HPLC-MS/MS method for determination of evodiamine enantiomers in rat plasma and brain was developed and verified. Satisfactory enantioseparation of evodiamine was achieved on a Chiralcel OD-RH column with the mobile phase of acetonitrile-water (70:30, v/v) at a flow rate of 0.6 mL/min. The analytes were measured under multiple reactions monitoring (MRM) mode with m/z 304.3→134.3 for evodiamine and m/z 289.3→97.1 for testosterone (IS) using electrospray ionization source (ESI) in the positive ion mode. The method was validated and fulfilled the requirements of bioanalysis. It was successfully applied to study the stereoselectivity of evodiamine in pharmacokinetics and the ability across the blood-brain barrier in rats. After oral and intravenous administration of racemic evodiamine to rat, the area under the concentration-time curve of R-evodiamine was noticeably 1.70 and 1.33 times higher than those of S-evodiamine, respectively. Furthermore, while there was no significant difference in the B/P values, the concentration of R-(-)-evodiamine in the brain was approximately 1.31 times greater than that of S-(+)-evodiamine. The results indicated that evodiamine enantiomers exhibited significant stereoselectivity in pharmacokinetics after oral administration and intravenous administration. While the two enantiomers showed no significant stereoselectivity in the ability across the blood-brain barrier. These findings provided new insights into the development of a single enantiomer of evodiamine as a potential drug candidate for the treatment of CNS diseases.