Metabolism-based Structure Modification And Biological Activity Study Of Triptolide

In the long history between human and disease resistance,traditional Chinese medicine plays an important role and have become a huge treasure for drug development.Triptolide is derived from Chinese medicine Tripterygium wilfordii,a tri-epoxy diterpene compound with broad-spectrum and high-efficiency anti-tumor activity.It can inhibit the proliferation of many kinds of tumor cells.Its anticancer activity is even stronger than paclitaxel,doxorubicin,cisplatin and other first-line anticancer drugs.However,the pharmacologists' development of triptolide was ceased in the clinical stage,which is not only due to the severe side effect of triptolide,but also the poor drug-like properties have become an important reason to limit its clinical application in the treatment of cancer,especially in its rapid clearance rate in vivo and short half-life.This may lead to frequent dosing,which in turn increases the patient's physical,mental,and economic burden.Therefore,the aim of this study is to use the traditional Chinese medicine active ingredient triptolide as a lead compound to modify its structure.Design and computer screen and synthesis triptolide derivatives,and study the in vitro antitumor activity of different triptolide derivatives against different tumor cells and in vitro metabolic stability and kinetics of enzymatic reactions.A study of relationship between structure and activity of triptolide has been provided,expecting to guidance for the rational design and synthesis of new drugs.Four parts of experimental process have been described as follow:1.Chemical synthesis of triptolide derivatives: Triptolide is used as a lead compound,and an effective drug group is introduced through structural modification to block or increase the metabolic sites.triptolide derivatives with good properties have been synthetized by computer screening2.Structure identification of derivatives: The newly synthesized triptolide derivatives were structurally identified by spectroscopic methods(1H NMR,13 C NMR).3.Study on anti-proliferative ability of triptolide and its derivatives in vitro: LNCa P,SW-480,KAT-10,and A549 cells were cultured.Different concentrations of drugs have been added to different cells for 24 h,48 h and 72 h.Cell viability and IC50 of each compound have been determined.4.In vitro metabolic stability of triptolide derivatives: The metabolic cofactors NADPH and UDPGA were added into the reaction system to initiate cytochrome P450 and UGT enzyme-mediated biotransformation.The substrate enzyme elimination method was used to determine the apparent enzyme kinetic parameters of triptolide derivatives in human liver microsomes.In this summary,Triptolide derivative TP1-3 was obtained during the experiment,and its structure was confirmed by hydrogen spectroscopy and carbon spectroscopy.In vitro anti-proliferation experiments showed that the triptolide derivative TP2 had inhibition on LNCa P,SW-480,KAT-10,and A549 cells.Among them,TP1 had less inhibition on each cells.The TP3 can effectively inhibit LNCa P,A549,but its effect is lower than TP2.The metabolic stability of TP1 and TP2 has been significantly improved compared to triptolide,and the half-life has been extended,indicating that this structural modification has a significant effect on the metabolic stability of triptolide.