Novel biotransformation processes of dihydroartemisinic acid and artemisinic acid to their hydroxylated derivatives by two plant cell culture systems

Novel biotransformation processes of dihydroartemisinic acid (1) and artemisinic acid (2) to their hydroxylated derivatives were investigated using the cell suspension cultures of Catharanthus roseus and Panax quinquefolium crown galls as two biocatalyst systems. Five biotransformation products, 3-alpha-hydroxydihydroartemisinic acid (3), 3-beta-hydroxydihydroartemisinic acid (4), 15-hydroxy-cadin-4-en-12-oic acid (5), 3-alpha-hydroxyartemisinic acid (6) and 3-beta-hydroxyartemisinic acid (7), were isolated by chromatograph methods and identified by the analysis of H-1 NMR, C-13 NMR, and ESI-MS spectra. Compounds 3-5 were obtained for the first time by biotransformation process. It was also the first time to transform artemisinic acid to yield epimeric 3-hydroxy artemisinic acids in plant cell culture system. The biocatalyst system of C. roseus cell cultures showed a great capacity of regio- and stereo-selective hydroxylation in allyl group of the exogenous substrates. The results also showed that the biocatalyst system of P. quinquefolium crown galls possessed the ability to hydroxylate propenyl group of exogenous substrates in a regio- and substrate-selective manner. Furthermore, the in vitro antitumor activity of the hydroxyl products was evaluated by MTT assay. The result indicated that a-hydroxyl products possessed stronger antitumor activity than alpha-hydroxyl products against the HepG2 and GLC-82 cell lines. (C) 2010 Elsevier Ltd. All rights reserved.