Rapid Identification of Aldose Reductase Inhibitory Compounds from Perilla frutescens

الملخص EN

The ethyl acetate (EtOAc) soluble fraction of methanol extracts of Perilla frutescens (P.

frutescens) inhibits aldose reductase (AR), the key enzyme in the polyol pathway.

Our investigation of inhibitory compounds from the EtOAc soluble fraction of P.

frutescens was followed by identification of the inhibitory compounds by a combination of HPLC microfractionation and a 96-well enzyme assay.

This allowed the biological activities to be efficiently matched with selected HPLC peaks.

Structural analyses of the active compounds were performed by LC-MSn.

The main AR inhibiting compounds were tentatively identified as chlorogenic acid and rosmarinic acid by LC-MSn.

A two-step high speed counter current chromatography (HSCCC) isolation method was developed with a solvent system of n-hexane-ethyl acetate-methanol-water at 1.5 : 5 : 1 : 5, v/v and 3 : 7 : 5 : 5, v/v.

The chemical structures of the isolated compounds were determined by 1H- and 13C-nuclear magnetic resonance spectrometry (NMR).

The main compounds inhibiting AR in the EtOAc fraction of methanol extracts of P.

frutescens were identified as chlorogenic acid (2) (IC50 = 3.16 μM), rosmarinic acid (4) (IC50 = 2.77 μM), luteolin (5) (IC50 = 6.34 μM), and methyl rosmarinic acid (6) (IC50 = 4.03 μM).