| Prunella vulgaris L.(Labiatae) is widely famous for its various pharmacological research, moreover, it was confirmed that Prunella vulgaris has good anti-tumor effect. Nowadays, the aqueous extract from Prunella has been used in clinical. However, the anti-tumor basis and its molecular mechanism are not clear. In addition because of the complexity of component, mostly current preparations of Prunella were decoction or the simple formulation of its extract. And so for the above reasons, the assessment and control of quality of preparations were still difficult. In this study, three compounds with synergistic anti-tumor effects were identified from Prunella by using network pharmacology approach, and then were prepared to a multi-component preparation by solid dispersion technique. Studies on detection and evaluation in vitro and in vivo of the preparation were also conducted. This thesis focused on the identification of antitumor compounds of Prunella and the study of insoluble multi-component preparation mainly was divided into the following four parts: Part I ReviewIn this section, the information focus on anti-tumor drug trend, network pharmacology in traditional Chinese medicine, solid dispersion technologies and Prunella vulgaris was reviewed. Firstly, the analysis of anti-tumor drugs approved by FDA shown that multi-targets anti-tumor drugs has been a hotspot. Secondly, traditional Chinese medicine was used as a model to study multi-targets drugs because of their multi-targets synergistic action. Simultaneously, in this part, the research processes of traditional Chinese medicine by using network pharmacology approach were summarized, which provided methodological support for this study. Fourthly, the data about anti-tumor of Prunella were collected, which provided the data basis; At last, the present situation about solid dispersion technologies and traditional Chinese medicine preparation were generalized, which provided the theory basis for insoluble multi-component preparation. This section was the summary of ideas and methods as the important reference to carry out the study. Part II Multi-level interaction network based identification of anti-tumor active compounds from Prunella Vulgaris LIn this section, the Prunella anti-tumor network has been established consist of compounds, biomolecular(targets) and tumors, and the connectivity analysis and density-based network module analysis were used to screen anti-tumor active compounds. Because of the redundancy in the network, in order to select active compounds as few as possible, pathway analysis was carried out to further identify the active compounds based on the criterion that choosing the less compounds to affect the most anti-tumor related pathways. The synergies and the best proportion of the selected three compounds have been confirmed by MTT cell experiments. The results showed that the combination of Rutin, Quercetin and Ursolic acid could modulate most of the anti-tumor pathways and the inhibition rate reached to 96% when the dosage of all three compounds was 4μg. It suggested that Rutin, Quercetin and Ursolic acid performed synergistic anti-tumor effect. Part III The prescription design of insoluble multi-compoundsIn this section, the insoluble multi-component preparation of the identified compounds by network pharmacology approach was designed in order to improve the release rate of the three compounds and to make sure that all the compounds would smoothly release in a longer time. Here, the strategy for the preparation consists of two parts: first was to improve the cumulative release rate by velocity release preparation; second was to adjust the release process by choosing the appropriate release carriers. And there were two prescription design ideas. On the one hand, the three compounds was firstly solubilized and then prepared to sustained release solid dispersion respectively, finally mixed together. On the other hand, the three compounds were prepared as one whole like the simple prescription. At last, compared with the release rate of the prescriptions, the best prescription was determined: the three compounds were dissolved by ethanol together, PVPK30 and EC were choosing as the carriers with the rate of compounds and carries was controlled as 1:6:3, the action time was conformed as 30 min. The similarity factor between release curve of the three compounds all were over 50, which indicated that the three compounds from the preparation could release smoothly in vitro. The DSC and IR spectroscopy were used to verify that the insoluble multi-compounds preparation made by the above method was real solid dispersion. Part IV The study on in vivo pharmacokinetics of the insoluble multi-compounds preparation in ratsIn this section, the study on in vivo pharmacokinetics of the insoluble multi-compounds preparation using SD rats as animal models was carried out. The analytical method of the three compounds was established to investigate the release of every compound in vivo. The results showed that the peak concentration of Rutin, Quercetin and Ursolic acid were 2.920±0.987μg/m L,2.486±0.768μg/mL and 1.941±0.852μg/m L, which were all above the peak concentration of the three compounds in the control(physical mixture of three compounds, 0.933±0.135μg/m L, 0.883±0.1002μg/m L,0.919±0.221μg/m L), the half-life of Rutin, Quercetin and Ursolic acid were 9.219 h,11.887 h and 17.086 h, which were all prolonged 5.278 h,7.426 h,11.155 h than that of control, respectively. Simultaneously, the relative bioavailability of Rutin, Quercetin and Ursolic acid were 574.92%, 357.77% and 306.03%. Therefore the insoluble multi-compounds preparation has made the ingredients exist longer time in vivo and improved theirs’ bioavailability.Above all, it is feasible to identify the active compounds from traditional Chinese medicine through the network pharmacology approach, and to prepare multi-component preparation to promote the development of the modernization of the traditional Chinese medicine. |
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