Synthesis And Anti-inflammatory Activities Of 1-methylhydantoin Cinnamic Acid Derivatives

Inflammation is closely related to a variety of diseases that humans suffer.Current treatments for inflammation mostly use non-steroidal anti-inflammatory drugs.However,long-term high-dose use of such drugs will cause serious gastrointestinal adverse reactions.Therefore,it is of great significance to develop new drugs with low toxicity and high efficiency.Studies have shown that cinnamic acid derivatives and 1-methylhydantoin are small molecule compounds with anti-inflammatory potential.In this paper,1-methylhydantoin and cinnamic acid compounds were used as the basic skeleton based on the principle of drug combination,and the target compounds were obtained by synthesizing 1-methylhydantoin cinnamamide structure,so as to carry out anti-inflammatory research and screen out compounds with anti-inflammatory effects.In this project,a large amount of 1-methylhydantoin was prepared based on the preliminary research in the laboratory,and with 1-methylhydantoin and cinnamic acid compounds as the basic skeleton,five 1-methyl hydantoin cinnamic acid derivatives were initially synthesized through acetylation reaction,acid chloride reaction and amide reaction.The target compounds were characterized by ¹H NMR,¹³C NMR,IR,UV,MS.In the optimization of the synthesis process,the amount of acetic anhydride in the acetylation reaction;the amount of solvent,temperature and SOCl₂ in the acid chloride reaction;the reaction time and the amount of 1-methylhydantoin in the amide reaction were investigated.The results showed that when the molar amount of acetic anhydride was 3 times that of coumaric acid and ferulic acid,the acetylation reaction was complete and the yield was high;In the acid chloride reaction,all five cinnamic acid derivatives could be converted into acid chlorides at room temperature,when CHCl₂ was the solvent and SOCl₂ was 1.1 times that of carboxylic acid;the optimal reaction time of the five compounds（A-E）in the amide reaction was:6 h,6 h,4 h,8 h,8 h,and the optimal molar ratio of acid chloride to 1-methylhydantoin was:1:0.9,1:0.9,1:1,1:0.9,1:1.When investigating anti-inflammatory activity,firstly,the purity of the five compounds was determined using the high-performance liquid.Combining the chromatogram and chromatographic peak information,it can be seen that the content of the five compounds is more than 95%,which provides a good basis for subsequent activity experiments.Secondly,CCK-8 method was used to detect the cytotoxicity of1-methylhydantoin cinnamic acid derivatives to RAW 264.7 cells,and LPS was used to induce RAW 264.7 cells to form an in vitro cell inflammation model.The content of NO,TNF-αand IL-1βin the cell supernatant was used to evaluate the in vitro anti-inflammatory activity of 1-methylhydantoin cinnamic acid derivatives.The results showed that when the concentration of compound A,compound C,and compound E was lower than 20μM and the concentration of compound B and compound D was lower than 80μM,the cell survival rate was above 95%.Compound B and compound D could significantly inhibit the release of NO,reduce the secretion of TNF-αand IL-1β,and compound C on TNF-αproduction also had an inhibitory effect.The in vivo anti-inflammatory activity of five compounds was evaluated by establishing an animal model of xylene-induced ear swelling.The results showed that compounds A,B,C,D,and E could alleviate xylene-induced ear edema in mice,and they were dose-dependent,compound E had the most significant effect,and the inhibition rate of ear swelling was as high as 52.08%under the action of high dose.Finally,using molecular docking technology to predict compounds with the potential to selectively inhibit COX-2,computer simulation results showed that the affinity of compounds B,C,D,E to COX-2 was higher than that of COX-1,and had the potential to selectively inhibit COX-2.