Chemical Constituents And Biological Activity From The Flowers Of Hemerocallis Fulva

Objective:（1）A systematic study on the chemical components of the whole flower of Hemerocallis fulva was carried out to clarify the main chemical components of Hemerocallis fulva and its medicinal material basis,so as to provide a basis for the formulation of the quality standard of Hemerocallis fulva.（2）The bioactivity of 95%ethanol extract of dried whole flower of Hemerocallis fulva was evaluated,and the corresponding pharmacodynamic material basis of heat clearing,detoxification and anti-inflammatory effects of the plant was explained.Methods:（1）The compounds were isolated and purified by normal phase silica gel column chromatography,thin layer preparative chromatography,reverse ODS column chromatography,MCI column chromatography,gel column chromatography（Sephadex LH-20）,semi preparative high performance liquid chromatography and recrystallization.Using 1D NMR（1H-NMR,13C-NMR）,two-dimensional nuclear magnetic resonance（HSQC,HMBC,NOESY）and mass spectrometry and other modern spectroscopy techniques,combined with published literature,the data were compared.Analyze and determine the structure of the compound.（2）In vitro cell hemolysis method was used to determine the anti complement activities of polar extraction parts and some monomer compounds of Hemerocallis fulva.（3）Based on the principle of network pharmacology,active molecules were screened and related targets were obtained through swissadme database;The target data were imported into CTD database for analysis to obtain relevant diseases,the KEGG pathway was analyzed by David online analysis tool,and the relevant network model diagram was drawn and analyzed by Cytoscape software.Results:（1）22 monomers were isolated and identified from the n-butanol part of 95%ethanol extract of the flowers of Hemerocallis fulva,including 4 new compounds,5-O-（1-O-pcoumaroylglycerol）jatropham（1）,1-O-pcoumaroylglycerol（2）,1-O-caffeoylglycerol（3）,sinapic acid（4）,3-hydroxy-4-methoxycinnamic acid（5）,ferulic acid（6）P-hydroxycinnamic acid（7）,hawthorn leaf element B（8）,imperylone A（9）,1,3-differoylglycerol（10）,perlolyrine（11）,pyrrolealkaloid（12）,5-（butoxymethyl）-1H-pyrrole-2-carbaldehyde（13）,5-O-butanejatropham（14）,5-O-methyljatropham（15）,（R）-jatropham（16）,5,5’-oxydi（3-methyl-3-pyrrolin-2-one）（17）quercetin（18）,（+）-pinoresinol（19）,methylsuccinic acid（20）.（2）The classical anti complement activity test method-cell hemolysis method was used to determine the anti complement activity of various polar extraction parts and some monomer compounds of the flowers of Hemerocallis fulva.Among them,the ethyl acetate extraction part of the flowers of Hemerocallis fulva has the most significant activity,CH₅₀ value is 0.016mg/ml,n-butanol extraction part has strong activity,CH₅₀value is 0.03mg/ml,petroleum ether extraction part has weak activity,CH₅₀ value is 0.06mg/ml,and the water extraction part has almost no activity,CH₅₀value is 0.291mg/ml.Monomeric compounds 3,8,9,12and 16 showed anticomplement activity,and their CH₅₀ values were 1.053 m M,1.422 m M,1.281 m M,0.614 m M and 1.433 m M respectively.（3）Nine compounds and 289 related targets were screened from the chemical constituents of Hemerocallis fulva.HSP90AA1（heat shock protein）and SRC（tyrosine specific protein kinase）are the key targets of Hemerocallis fulva flower in the treatment of tumor,inflammation and other diseases.The relationship between compound targets and related diseases and signal pathways shows that Hemerocallis fulva can play a good role in tumor,digestive system and immune system related diseases.Conclusions:（1）Among the 20 monomeric compounds isolated and identified from the n-butanol part of 95%ethanol extract of Hemerocallis fulva flower,3 were new structural compounds（compounds 1,13 and 14）.Among the structural types of compounds,10 are phenolic compounds（compounds 1～10）,7 are alkaloid compounds（compounds 11～17）,1 is flavonoid compound（compound 18）,1 is lignan compound（compound 19）and 1 is fatty acid compound（compound 20）.（2）Except compound 18,other compounds were isolated and identified from this plant for the first time,and also from Hemerocallis for the first time.（3）The content of phenols and alkaloids in Hemerocallis fulva flower is large.It is speculated that phenols and alkaloids are the pharmacodynamic material basis corresponding to the antipyretic,detoxifying and anti-inflammatory effects of Hemerocallis fulva flower.（4）The n-butanol parts of Hemerocallis fulva flower showed anticomplement activity,which is the pharmacodynamic material basis of the anti complement activity of Hemerocallis fulva flower.Monomeric compounds 3,8,9,12 and 16 constitute the anti complement active components of Hemerocallis fulva flower.（5）The results of network pharmacological analysis showed that the related targets of Hemerocallis fulva active components were related to tumors,immune system related diseases and signal pathways.The pharmacodynamic material basis of Hemerocallis fulva.