Synthesis And Anti-tumor Activity Studies Of Phenethyl Cinnamate Analogues

It is well known that propolis is a natural medicine. It possesses the ettect of immune regulation and other activities. CAPE is a natural ingredient extracted from propolis. It is known to exhibit antiinflammatory, antioxidant, antineoplastic and immunomodulatory properties, especially the antioxidant property. Based on this, we have synthesized a series of cinnamate analogues and discussed their antitumor activity.This paper is made up of four parts. The first chapter is the reviews; The second chapter describes the synthesis of caffeic acid phenethyl ester analogues; The third chapter represents the pharmacology of cinnamate analogues; The last chapter summarizes the conclusions of the research. The structure of CAPE is illustrated in Fig.1-1. Chapter1The ReviewsThis chapter sums up the research progress of CAPE and its analogues by reviewing more than60pieces of literatures.The first section introduces mainly the pharmacological activity of CAPE. Its activities were widely noted since it has been obtained from the extracts of the propolis; The second section summarizes the methods that how to synthesize CAPE. The last section puts emphasis on discussing the pharmacological activities of cinnamate analogues and their structure activity relationships. Chapter2The synthesis of cinnamate analoguesThe first section of this chapter provides the method to prepare compounds1-11. They are3-phenyl-acrylicacidphenethylester,3-(3,4-dimethoxy-phenyl)-acrylic acid phenethyl ester,3-(3,4,5-trimethoxy-phenyl)-acrylic acid phenethyl ester,3-(4-methoxy-phenyl)-acrylic acid phenethyl ester,3-(2,3-dimethoxy-phenyl)-acrylic acid phenethyl ester,3-p-tolyl-acrylicacidphenethylester,3-(2-methoxy-phenyl)-acrylic acidphenethylester,3-(3-methoxy-phenyl)-acrylic acid phenethylester,3-(2,5-dimethoxy-phenyl)-acrylic acid phenethyl ester,3-(2,3,4-trimethoxy-phenyl)-acrylic acid phenethyl ester and3-(4-dimethylamino-phenyl)-acrylic acid phenethyl ester. The second part mainly describes the route that synthesizes compounds12-17. They are3-(3-hydroxy-phenyl)-acrylicacidphenethylester,3-(4-hydroxy-phenyl)-acrylic acid phenethyl ester,3-(4-hydroxy-3-methoxy-phenyl)-acrylic acid phenethyl ester,3-(2-hydroxy-phenyl)-acrylic acid phenethyl ester,4-hydroxy-3-methoxy-benzoic acid phenethyl ester and4-hydroxy-benzoicacidphenethylester. Chapter3The experiments of cinnamate analogues’pharmacological activitiesThis part presents the antitumor activity about the raw material and cinnamate analogues. Firstly, the raw material and seventeen caffeic acid phenethyl ester analogues whose antiproliferative effects were evaluated toward Bel-7402, MCF-7and Hela human cancer cell lines; Secondly, we find that compound14is the most effective compound and its antitumor activity is evaluated toward HepG2human cancer cell lines. Besides, the apoptosis route was determined by flow cytometry. Then, based on the activity screening, overall observation on the effect of the compound14was made by establishing H22mice tumor models. What’s more, using pathological section dyeing with HE to find further progress on antitumor activity.Chapter4Summary and DiscussionBased on that propolis possesses a broad spectrum of biological and pharmacological properties and that CAPE is derived from propolis extract, this paper presents the method of synthesis of seventeen cinnamate analogues. Three of them are first synthesized. Their antiproliferative effects were evaluated toward Bel-7402and MCF-7cancer cell lines by MTT method. The following compounds are active:3-hydroxy-benzoic acid phenethyl ester,4-hydroxy-3-methoxy-benzoic acid phenethyl ester,3-(4-hydroxy-phenyl)-acrylic acid phenethyl ester,3-(3-hydroxy-phenyl)-acrylic acid phenethyl ester,3-(2-hydroxy-phenyl)-acrylic acid phenethyl ester,3-(4-hydroxy-3-methoxy-phenyl)-acrylic acid phenethyl este and CAPE. Their activity increased in turn. For the Bel-7402cell, the IC50values respectively are>200μmol·L-1,101.6μmol·L-1,92.87μmol·L-1,67.11μmol·L-1,58.55μmol·L-1,51.98μmol·L-1,42.26μmol·L-1; for the MCF-7cell, the IC50values respectively are81.26μmol·L-1,80.13μmol·L-1,55.67μmol·L-1,51.36μmol·L-1,44.59μmol·L-1,41.87μmol·L-1,34.6μmol·L-1. Some conclusions were obtained. The a,(3-unsaturated double bond and the phenolic hydroxyl group in caffeic acid phenethyl ester were essential for the antitumor activities. Besides, the cytotoxic properties of these compounds were determined by the methoxy groups on the phenyl ring and the location of aromatic hydroxyl groups. The A ring with the adjacent hydroxyl could enhance its effect in antitumor function. Besides, the cytotoxic properties of these compounds were determined by the methoxy groups on the phenyl ring and the location of aromatic hydroxyl groups. The A ring with the adjacent hydroxyl could enhance its effect in antitumor function. For the human Hela cancer celles,3-(2-hydroxy-phenyl)-acrylic acid phenethyl ester exhibits the best activity, and its IC50value is34.41μmol·L-1.Using the method of cell activity, the most effective compound was found and it was3-(4-hydroxy-3-methoxy-phenyl)-acrylic acid phenethyl ester. The percentage of apoptosis cells was detected by flow cytometry and the percentage is66.4. After its cytotoxicity in vitro by MTT assays, the anti-tumor effects on H22bearing mice in vivo were tested. The positive drug is cyclophosphamide and the anti-titumor rate of3-(4-hydroxy-3-methoxy-phenyl)-acrylic acid phenethyl ester is48.56%when using the concentration of30mg/kg. At last, using pathological section dyeing with HE to explore the mechanisms of pathology. These results may be of great benefit to people in screening new anticancer drug candidates.