Construction and Screening of Fractional Library of Radix et Rhizoma for the Rapid Identification of Active Compounds against Prostate Cancer.

Efficient screening of anticancer agents is in urgent need to develop new drugs that combat malignant tumors and drug resistance. In this study, a combined strategy composed by solvent partition and HPLC fractionation was developed to generate an herbal fraction library of Radix et Rhizoma to quickly and efficiently screen anticancer agents. All library entries are directed into 96 well plates which are well mapped with HPLC chromatograms. The cell proliferation assay revealed seven active subfractions. Then, the major active ten peaks in these subfractions were prepared and isolated by semipreparative HPLC, and their inhibitory activities against prostate cancer cells were then tested at the same concentration level, leading to the identification of several active compounds. In addition, the structures of compounds arucadiol (), 15,16-dihydrotanshinone I (), methyl tanshinonate (), cryptanshinone (), 1,2-dihydrotanshinquinone I (), and tanshinone IIA () were characterized by mass spectrometry and X-ray crystallographic analysis, and they were confirmed to be active in suppressing prostate cancer cell proliferation at 7.5 or 15 g/mL, among which, the minor compounds , , and showed higher activities than and . This study provided a rapid strategy of identifying new anticancer agents in Radix et Rhizoma, which can be applied in other herbal medicines.