Studies On The Constituents From The Fruits Of Evodia Rutaecarpa And Biological Activities

Objective:This dissertation describes studies on chemical constituents of 70%ethanol extract of the fruits of Evodia rutaecarpa and their various biological activities including hypoglycemic effect,anti-bacterial effect,and antitumor activity.Methods:The fruits of Evodia rutaecarpa was extracted with 70%Et OH.The extracts were combined and evaporated under reduced pressure to yield the aqueous residue.The aqueous residue was diluted with H₂O and sequentially partitioned with petroleum ether,Et OAc,and n-Bu OH.The compounds were isolated from the Et OAc extract by variety of chromatographic techiques such as silica gel column,MPLC,and HPLC.Their structures were elucidated on the basis of a variety of spectroscopic methods such as ¹H NMR,¹³C NMR,2D NMR,X-ray crystallography and MS,etc.Biological activities of isolated compounds were evaluated through their inhibitory activity of protein tyrosine phosphatase 1B（PTP1B）,α-glucosidase,bacteria proliferation and cytotoxicity.Results:Twenty five constituents（1-25）were isolated and identified from the70%Et OH extract from Evodia rutaecarpa on the basis of 1D NMR,2D NMR,HRESIMS,X-ray crystallography and comparison of experimental CD spectra with the calculated ECD spectra.The new compounds were named evolide A（1）,evolide B（2）,10-methoxygoshuyuamide-Ⅱ（3）and the known compounds were identified as goshuyuamide-Ⅱ（4）,（S）-7-hydroxysecorutaecarpine（5）,3-hydoxyrutaecarpine（6）,rutaecarpine（7）,evodiamine（8）,7β-hydroxyrutaecrpine（9）,dehydroevodiamine（10）,limonellone（11）,7-hydroxy-1-benzopiran-2-one（12）,limonin（13）,evodol（14）,evodirutaenin A（15）,isorhamnetin（16）,trans-ferulic acid（17）,trans-p-coumaric acid（18）,syringaldehyde（19）,vanillic acid（20）,protocatechuic acid ethyl ester（21）,cis-caffeic acidethyl ester（22）,（E）-ethyl 3-（3,4-dihydroxyphenyl）acrylate（23）,affeic acid methyl ester（24）,and（2E,6S）-6-hydroxy-2,6-dimethylocta-2,7-dienoic acid（25）,and compounds（1-3）were new compounds.The bioactivities of compounds（1-25）were evaluated on several pharmacological models.Compounds 3,4,and 6 could inhibit the PTP1B enzyme activity in a dose-dependent manner.The IC₅₀values were 75.83±14.7,60.61±8.31,and 16.19±0.39μM,respectively.Compounds 3,4,6,and 8 could inhibit theα-glucosidase activity in a dose-dependent manner.The IC₅₀values were 23.92±0.81,22.11±2.53,8.77±0.14,and16.21±1.39μM,respectively.By CCK8 method,the in vitro cytotoxicity activities against cancer cell lines of compounds（1-25）were assayed.Compound 3 exhibited significant cytotoxic activities against SHSY-5Y,the IC₅₀values were 24.7μM.Compound 8 exhibited significant cytotoxic activities against Hep G-2,A498 and SHSY-5Y,the IC₅₀values were<1μM.Compounds 3,8,and 9 displayed moderate inhibitory effects on Bacillus cereus,with MIC values 50μM,10μM and 25μM,respectively.Conclusion:This dissertation investigated chemical constituents of Evodia rutaecarpa and their bioactivities.The results indicated that there were various chemical constituents in Evodia rutaecarpa.These isolates are categorized as two sesquiterpenoids（1-2）,twenty two alkaloids（3-11）,one coumarin（12）,three limonin analogues（13-15）,one flavonoid（16）,and nine others（17-25）.They displayed various biological activities including anti-bacterial effect,antitumor activity,and hypoglycemic effect,which could develop the application of Evodia rutaecarpa.