Discovering And Screening Of Withanolides And Physalins With Anti-inflammatory And Cytotoxic Activities

【Subject basis and Research content】The control and treatment of inflammation and tumors remains a difficult and hot topic in medical research.Discovering anti-inflammatory and cytotoxic candidate molecules from natural products is a current mainstream and feasible pathway.of new drug discovery.Withanolide is a kind of 28 carbon steroid natural product derived from ergostane skeleton.Its structural feature is that C-22 and C-26 or C-23 and C-26are linked in the side chain to form aδ-orγ-Ester;Physalin is a kind of ergostane features a basic mother nucleus 13,14-cracked-16,24-cycloergosterol,named for its bitter taste.Some of the compounds of these two classes exhibit significant activity against tumors and inflammation.Our previous investigations and studies have found that these two components are present in two kinds of Solanaceae plants,Nicandra physaloides and Physalis alkekengi L.In order to further obtain structurally diverse withanolides and physalins,to lay the material foundation for screening natural products with anti-inflammatory and cytotoxic activities,and to provide candidate molecules for the research and development of innovative anti-inflammatory and cytotoxic drugs,we first carried out a systematic separation of N.physaloides and P.alkekengi L.to obtain withanolides and physalins and identified their structures by mass spectrometry,nuclear magnetic resonance and other spectroscopy techniques,Then we screened for their anti-inflammatory and cytotoxic activity usinge an in vitro cell model.This study will lay the foundation for the development of anti-inflammatory and cytotoxic innovative drugs with independent intellectual property rights.【Methods and Results】In this experiment,38 compounds were isolated and identified by extraction and various separation methods from plants of P.alkekengi L.and N.physaloides.In total,19 compounds（compounds 1-19）were isolated from N.physaloides and were identified by analytical spectroscopy data,all of which were withanolides,of which 3were new compounds.They are:nicphysatone A（1*）,nicphysatone B（2*）,nicphysatone C（3*）,Altogether 19 compounds（compounds 20-38）isolated and identified from P.alkekengi L.,including 6 physalins,and 13 withanolides.There are three new compounds among them,they are alkekenginin C（20*）,physapubescin M（21*）,physapubescin N（22*）,respectively.The effects of different compounds on the proliferation of tumor cells A-549 and MCF-7 were detected by CCK-8 method.The results showed that physapubescin M（21*）,physalin A（28）,physapubescin（29）,physapubescin K（30）and alkekenginin F（33）inhibited the proliferation of A-549and MCF-7 cells with IC₅₀ values of 3.96-27.10μM;The macrophage RAW 264.7model was stimulated by lipopolysaccharide（LPS）to detect the effects of different compounds on the release of NO.The anti-inflammatory activity was observed.The results showed that physapubescin M,physapubescin K and alkekenginin F had strong anti-inflammatory activity.The IC₅₀ values were 1.70-3.00μM.【Conclusion】This project mainly studies the compounds and activities of two kinds of compounds,namely,withanolides and physalins.A total of 32 withanolides and 6physalins were isolated from N.physaloides and P.alkekengi L.,of which 6compounds were new withanolides.The project lays a preliminary foundation for further obtaining structurally diverse withanolides and physalins,and developing a natural product with independent intellectual property rights.In the activity investigation,the compounds physapubescin M,physapubescin K and alkekenginin F have both cytotoxic and anti-inflammatory activity,and physapubescin M is withanolide which isolated from the P.alkekengi L for the first time.The project provides candidate molecules for the research and development of innovative anti-inflammatory and cytotoxic drugs.