Synthesis And Anti-inflammatory Activity Evaluation Of Resveratrol-based Phthalide Derivatives

Inflammation is a clinically common disease that has received widespread attention as a basis for the pathogenesis of many major diseases.Inducement and pathogenesis of inflammation are complex.At present,the drugs for inflammation therapy are mainly non-steroidal anti-inflammatory drugs,but these drugs usually cause serious adverse reactions to patients.As a result,a lot of research are committed to look for anti-inflammatory drugs with more efficient and less side effects.In recent years,phthalide and resveratrol have been widely studied as important natural products,and have been widely used in the design of drugs because of their extensive physiological activities such as anti-oxidation,anti-inflammatory and anti-tumor.In this paper,a series of phthalide-resveratrol heterozygous derivatives were designed.The pyrophyll was used as the starting material to synthesize a total of 52compounds in two series.The synthesized compounds were characterized by nuclear magnetic resonance and high resolution mass spectrometry,and some compounds were confirmed by X-ray single crystal diffraction.LPS-induced RAW 264.7 cell line was used to establish an inflammatory cell model in vitro and explored the anti-inflammatory activity of the synthesized compounds.The effects of compounds on the release of NO from RAW264.7 cells induced by LPS were tested by Griess method.The results have shown that all synthetic resveratrol-phthalide hybrid derivatives have different degrees of inhibition on the release of NO,and the inhibition of compound D₂ is most potent(IC₅₀=0.76?M).The MTT assay demonstrated that all compounds were not significantly toxic to RAW264.7 cells when effectively exerting anti-inflammatory activity.Western blot demonstrated that compound D₂ has a good inhibitory effect on inducible nitric oxide synthase?iNOS?and cyclooxygenase-2?COX-2?,and can effectively inhibit I?B and p65 proteins in NF-?B signaling pathway.It was found that compound D₂ can inhibit the expression of IL-1?and IL-18.The inhibitory effect of D₂ on NLRP3,ASC and Caspase-1 demonstrated that the anti-inflammatory effects of D₂ was mainly mediated by the blockade of NF-?B-NLRP3-ASC-Caspase-1 signaling pathway.