Design,Synthesis And Anti-hepatoma Activity In Vitro Of Winolide A Derivatives

Hepatocellular carcinoma?HCC?is one of the most common malignant tumors in the world.Its clinical features are concealment,rapid progress and poor prognosis,leading to the second highest mortality rate among all cancers in the world.The incidence of HCC in China is highest around the world,and the number of patients with hepatocellular carcinoma accounts for more than 50%of the global total.At present,sorafenib is the only one drug used in the treatment of patients with advanced liver cancer.However,sorafenib is prone to produce drug resistance and side effects.So,it is urgent to develop new effective anti-liver cancer drugs.Searching novel active compound from natural products is an important way to develop new drugs.In the previous study,we found that Winolide A inhibited the proliferation of hepatoma cells and induced the apoptosis of four hepatoma cells?SMMC-7721,QGY-7703,Huh-7 and HepG2?.In this study,the structural modifications were performed with Winolide A as the lead compound,aiming to obtain compounds with better activity and less toxicity.A total of 61 derivatives were synthesized using propiolic acid and cis-2-butene-1,4-diol as raw materials,followed by esterification,ene acetylene rearrangement,oxidation and amidation,.All derivatives were confirmed by mass spectroscopy,¹H-NMR,and ¹³C-NMR.Secondly,the experiments on inhibiting the proliferation of hepatoma cells and the toxicity on normal liver cells L02 were performed.The results showed that the activity of compound 2-3 against hepatocellular carcinoma cells was nearly 10-fold higher than that of the Winorlide A,and the IC₅₀ of the hepatoma cell line Huh-7 was1.81??,and the toxicity to the normal liver cell L02 was reduced.In addition,compounds 2-2 and 2-6 also displayed good activity on human hepatocellular carcinoma cell line Huh-7,IC₅₀ were 2.55??and 4.01??.The preliminary apoptosis mechanism was explored through apoptosis and western-blot experiments.The results showed that compounds 2-2,2-3 and 2-6significantly induced apoptosis of four hepatoma cells in a dose-dependent manner.2-3 had the best apoptosis-inducing effect on hepatoma cell line Huh-7;all three compounds could regulate the expression of caspase 3 and caspase 9 in four types of hepatocellular carcinoma cells,achieving the effect of apoptosis.Based on the above experimental results,derivative 2-3 can be further studied as a preferred compound.This study hopes to provide a new direction for the discovery of the anti-HCC candidate,and also provides some references for other natural products.