Design,Synthesis And Preliminary Activity Study Of 5-FU-HDAC Target Analogue

5-Fluorouracil(5-fluorouracil,5-FU)is an anti-metabolic drug that prevents tumor cell pyrimidine nucleotide formation and is mainly used for the treatment of colorectal cancer,gastric carcinoma,liver cancer etc.However,the clinical applications of 5-FU are subjected to great limitations due to its short plasma half-life,poor tumor selectivity,bone marrow suppression,intestinal toxicity and so on.Therefore,researchers have been working to develop fluorouracil derivatives with high efficacy and low toxicity as a new type of antitumor drug.In recent years,HDACs are promising nonkinase targets for therapeutic interventions intended to reverse aberrant epigenetic states associated with cancer.HDAC can mediate the change of nucleosome structure and regulates gene expression,it also participates in cell cycle progression and differentiation.HDAC can inhibit the acetylation of tumor cells,this process can condense DNA structure and prevent transcription,leading to cell differentiation and death.SAHA(Vorinostat),an HDAC inhibitor,was approved by the FDA for the treatment of cutaneous T cell lymphoma(CTCL)in 2006.A variety of HDAC inhibitors are currently in clinical studies,either individually or in combination with other anticancer drugs.HDACs are a type of Zn~+-dependent metalloproteinases located in nucleus,and their inhibitors contains the corresponding zinc-binding group(ZBG).Morevoer,the target of 5-FU is also located in the nucleus.In order to improve the liposolubility and side effects of 5-FU,we reviewed the progess of the structural modification of 5-FU and HDAC inhibitors,and found that the N-1 and N-3 of 5-FU and the para-and meta-of the phenyl ring of SAHA can be modified.We took the twin drug Benorilate and Progabide for example,with SAHA as carrier,designed and synthesized a 5-FU-SAHA analogue as a new antitumor prodrug.The biological activity of the compound was evaluated preliminarily.At last,Computer Aided Drug Design(CADD)was used to screen the HDAC inhibitors with high scores,which laid the foundation for the research of new types of HDAC inhibitors contained 5-FU.1.5-FU reacted with chloroacetic acid in potassium hydroxide solution at controlled temperature to prepare 5-fluorouracil-1-yl acetic acid.Methyl 8-chloro-8-oxooctanoate was synthesized form suberic acid by esterification,hydrolysis and acylation.Then it reacted with 4-nitrobenzylamine hydrochloride to prepare the intermediate compound WQ-04.The intermediate WQ-04 was reduced by iron powder to obtained compound WQ-05.Finally,target compound WQ-06 was synthesized by 5-fluorouracil-1-yl acetic acid condensated with WQ-05.The structures of all the above compounds were confirmed by nucler magnetic resonance and mass spectrometry.2.The biological activity of the compound was evaluated preliminarily by the comparison of 5-FU and SAHA.The result indicated that the in vitro antitumor activity of compound WQ-06 was higher than that of SAHA but lower than 5-FU,and suggested that compound WQ-06 does not affect the antitumor activity of 5-FU and SAHA after the modification.In the study of absorption,the absorption of WQ-06 was better than 5-FU,and indicated that the the modification of 5-FU can improve its liposolubility and targeting without affecting its anticancer activity.3.We selected HDAC as the target protein and the contained small molecule SAHA as the positive control.First,ompounds with ZBG were picked out through the UNITY module in SYBYL.Then the 30 compuonds with better docking score were selected as hits by the Surflex-dock module.We analyzed the binding modes of hits with HDAC that provided a theoretical basis for the research of novel lead 5-FU-HDAC target compounds.