Study On The Componunds Of Estrogen Receptor Regulating Effect In The Toddalia Asiatica

Rheumatoid arthritis?RA?is a systemic,autoimmune disease that belongs to the category of"bi syndrome"in traditional Chinese medicine and is one of the clinically refractory diseases.Toddalia asiatica[Toddalia asiatica?L.?Lam.]is the dried root bark.It has a long history of folk treatment for rheumatoid arthritis.However,the active ingredients and mechanism of action of toddalia asiatica to treat rheumatoid arthritis are still unclear.In the early stage of this project,we used chemmapper to search for potential targets of the coumarin compounds in the toddalia asiatica,Estrogen receptors account for the largest proportion of related targets.Based on this,this paper intends to explore whether estrogen receptors are potential targets for the treatment of rheumatoid arthritis by toddalia asiatica.In order to clarify active ingredients in the toddalia asiatica,this research used preparative liquid phase separation technology to separate the chemical composition of the ethyl acetate portion in the toddalia asiatica.while 14 compounds were isolated.Thereare10coumines.whichconsistof6-?3-Ethoxy-2-hydroxy-3-methylbutyl?-5,7-dimethoxy-2H-1-benzopyran-2-one?1?,coumarin?2?,toddanol?3?,toddanone?4?,toddaculin?5?,norbraylin?6?,braylin?7?,coumurrayin?8?,O-methylcedrelopsin?9?,artanin?10?;And there are 2 alkoloids:skimmianine?11?,?-fagarine?12?.One of phenylpropanoid:nelumol A?13?;One of the other compounds:tricyclo[7.2.0.0^2,4]undecane-5,8-diol,4,8,11,11-tetramethyl?14?.The new compounds is compound 1.The two-dimensional chromatographic separation technique was used to separate the n-butanol extraction part of the toddalia asiatica.As a result,4 compounds were obtained.Using high-resolution mass spectrometry,nuclear magnetic resonance,optical rotation and other spectroscopy techniques combined with reference data comparison methods to identify compounds.The structures of the identified compounds are?-?-toddalolactone2'-O-?-D-glucopyranoside?15?,?+?-toddalolactone 3'-O-?-D-glucopyranoside?16?,?-?-toddalolactone 2'-O-?-D-glucopyranoside?17?and hesperidin?18?.Compound 15 and16 are the new compounds,respectively.In order to verify the accuracy of the chemmapper database to predict the active target,this paper uses molecular docking technology to molecularly dock the isolated 10coumarin compounds with the crystal structure of the estrogen receptor,and screen for binding based on the binding free energy and inhibition constant Strong active ingredient.The results of molecular docking show that compared with other compounds,compounds2?5,7-dimethoxy-2H-1-benzopyran-2-one?,7?braeline?,9?O-methylcedrelopsin?and estrogen two different types of receptor proteins?1ERR,1QKN?have low free energy of binding,It is speculated that compounds 2,7,9 have strong estrogen receptor inhibitory activity.To our knowledge,this is the first report on the interaction between the coumarin component in the T.asiatica and estrogen receptors.And this is the first report on separating glycosides from T.asiatica using two-dimensional HPLC method.It lays a material foundation for further research and development of drugs with estrogen receptor modulation,which has important research value and application prospects.