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We present a paradigm, combining chemical profiling, absorbed components detection in plasma and network analysis, for investigating the pharmacology of combination drugs and complex formulae. On the one hand, the composition of the formula is investigated comprehensively via mass spectrometry analysis, followed by pharmacological studies of the fractions as well as the plasma concentration testing for the ingredients. On the other hand, both the candidate target proteins and the effective ingredients of the formula are predicted via analyzing the corresponding networks. The most probable active compounds can then be identified by combining the experimental results with the network analysis. In order to illustrate the performance of the paradigm, we apply it to the Danggui-Jianzhong formula (DJF) from traditional Chinese medicine (TCM) and predict 4 probably active ingredients, 3 of which are verified experimentally to display anti-platelet activity, i.e., (Z)-Ligustilide, Licochalcone A and Pentagalloylglucose. Moreover, the 3-compound formulae composed of these 3 chemicals show better anti-platelet activity than DJF. In addition, the paradigm predicts the association between these 3 compounds and COX-1, and our experimental validation further shows that such association comes from the inhibitory effects of the compounds on the activity of COX-1.

Combining Chemical Profiling and Network Analysis to Investigate the Pharmacology of Complex Prescriptions in Traditional Chinese Medicine