Synthesis Of Podophyllotoxin Derivatives And In Vitro Antitumor Activity

Malignant tumor,which were often refered to as cancer,are one of the diseases that pose a great threat to mankind in today’s society,with roughly one in six people dying of cancer each year worldwide,and therefore the search for effective anti-cancer drugs has been a major focus of scholarly attention.Podophyllotoxin is a kind of lignan compounds,which can be isolated from plants of Berberidaceae family.It has a variety of biological activities including anti-tumor,anti-inflammatory and anti-viral,with anti-tumor activity being the most significant.Structural modification of podophyllotoxin to enhance its anti-tumor activity and the discovery of new anti-tumor drug candidates have important theoretical and practical significance.In this study,a series of amino acid and organic acid ester podophyllotoxin derivatives were synthesized,evaluated for cytotoxicity against five human-derived cancer cells,and the 3D-QSAR analysis was carried out,and the mechanism of action was studied,which laid the foundation for drug development of podophyllotoxin compounds.The main research contents and results are as follows.1.Synthesis of podophyllotoxin derivatives.Ten kinds of Boc-amino acids and ten kinds of organic acids were used to carry out the esterification reaction with the hydroxyl group at the C-4 position of the C ring of podophyllotoxin,explore the reaction conditions.After further separation and purification,twenty kinds of ester derivatives V-13～V-32 were obtained,and their structures were confirmed by technical means such as spectroscopy.2.Evaluation of the in vitro antitumor activity of podophyllotoxin and its derivatives.The cytotoxicity of compounds V-13～V-32 and podophyllotoxin was evaluated against human prostate cancer cells PC-3M,human hemangioma endothelial cells Hem ECs,human lung cancer cells A549,human breast cancer cells MCF-7,human liver cancer cells Hep G2.Most of the compounds were found to have a good inhibitory effect on PC-3M and Hem ECs cells.Based on the IC₅₀ values obtained,the dominant compound V-17(IC₅₀=1.28±0.1n M)for PC-3M cells and V-31(IC₅₀=79.46±4.9n M)for Hem ECs cells were screened,conducted a follow-up experimental study on the mechanism of action.3.Preliminary exploration of the pro-apoptotic mechanism of compound V-17 in PC-3M cells.DAPI staining experiments showed that V-17 induced apoptosis in PC-3M cells,and cell morphological changes are observed;Colony formation assay showed that V-17 can inhibit the proliferation of PC-3M cells;Migration assay showed that V-17 can inhibit the migration of PC-3M cells;Flow cytometry analysis of the apoptosis rate found that the apoptotic rate of PC-3M cells treated with V-17 increased from 4.13%to 35.31%,which promoted cell apoptosis;Western blot experiments showed that its mechanism of action may be achieved by down-regulating p-PI3K,p-Akt and Bcl-2,and up-regulating Cleaved caspase-3 and Bax.4.Preliminary exploration of the pro-apoptotic mechanism of compound V-31 in Hem ECs.DAPI staining of Hem ECs cells showed that the contours of Hem ECs under V-31 treatment became indistinct and the nuclei were fragmented;Colony formation assay and migration assay showed that V-31 inhibited the proliferation and migration of Hem ECs;Flow cytometry analysis of apoptosis rate indicated that V-31 significantly promoted cell apoptosis,with the apoptosis rate increasing from 12%to 46.2%;Western blot analysis of related protein expression levels showed that V-31 may induce apoptosis in Hem ECs by down-regulating p-PI3K,p-Akt and Bcl-2,and up-regulating Cleaved caspase-3 and Bax.5.3D-QSAR study of podophyllotoxin derivatives.Using SYBYL-X 2.0 molecular simulation software,3D-QSAR studies were carried out with Co MFA and Co MSIA methods to obtain 3D isopotential maps of the Co MFA and Co MSIA models established between the compounds and the p M of PC-3M cells and Hem ECs cells.Based on this,the activity of unknown compounds can be predicted,which pointed the direction for the design and screening of new podophyllotoxin compounds.