Isolation And Identification Of Compounds From Inonotus Obliquus And Initially Study On Some Of Their Biological Activities

Inonotus obliquus is a medicinal and edible fungus belonging to Inonotus species in Hymenochaetaceae family.The sclerotia is the main medicinal part of I.obliquus.A large number of pharmacological experiments show that the extracts of I.obliquus functioned as anti-diabetes,anti-cancer,and immunity improving agents.The bioactivity assays of the compounds isolated from the extracts were mainly focused on hypoglycemic,anti-tumor and anti-oxidation.However,there are few studies on the anti-inflammatory,neuroprotective and cholinesterase inhibitory effects of the chemical components from I.obliquus.In order to enrich the chemical diversity and find more potential active compounds,as well as provide theoretical basis for the better utilization of I.obliquus resources and the development of related products.We performed a phytochemical study on the chemical components from ethyl acetate fraction of I.obliquus.And some of the compounds were tested for their cholinesterase inhibitory,cytotoxic.In addition,the effect of the compounds on NO production by LPS-induced RAW 264.7 mouse macrophages and the effect of H2O2-induced injury on SH-SY5Y nerve cells were detected.In our study,34 compounds were isolated from I.obliquus ethyl acetate fraction with a variety of column chromatography.Their structures were identified based on the NMR,MS,IR,and UV spectroscopic data,as well as by comparision with the literatures:2,3?,22?-trihydroxylanosta-8,24-dien(1),22,25-epoxy-lanosta-8-ene-3-one(2),inonotsutriol D(3),(20R,24S)-3?,25-dihydroxylanost-8-en-21,24-epoxy(4),(20S,24S)-3?,25-dihydroxylanost-8-en-21,24-epoxy(5),inonotsuoxide C(6),lanosterol(7),inotodol(8),3?-hydroxy-lanosra-8,24-dien-21-al(9),trametenolic acid(10),3?,21-dihydroxy-lanosta-8,24-diene(11),inoterpene D(12),betulinic acid(13),betulin(14),betulin 3?-caffeate(15),fuscoporianol A(16),lanosta-8,23E-diene-3?,22R,25-triol(17),inonotsutriol A(18),inonotsutriol C(19),inonotsuoxide A(20),monogynol A(21),inonotusol F(22),(3?,5?,6?,22E)-6-methoxyergosta-7,22-diene-3,5-diol(23),(22E)-5?,8?-epidioxyergosta-6,22-dien-3?-ol(24),3-hydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-3-(4-(3-hydroxypropanoyl)-2,6-dimethoxypheno xy)propan-1-one(25),4-((3-hydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-1-oxopropan-2-yl)oxy)-3-methoxyb enzoicacid(26),4-((3-hydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-1-oxopropan-2-yl)oxy)-3,5-dimethoxybenzo ic acid(27),inoscavin A(28),3,4-dihydroxybenzalacetone(29),2-(3',4'-dihydroxyphenyl)-1,3-benzodioxole-5-aldehyde(30),2,3-dihydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)propan-1-one(31),3-hydroxy-1-(4-hy droxy-3,5-dimethoxyphenyl)propan-1-one(32),1,3,5-trimethoxyb enzene(33),vanillic acid(34).Among them,triterpenoids 1-6 and phenolic acids 25-27 were new compounds,compounds 21,30 and 31 were isolated from I.obliquus for the first time.Some compounds were tested for cholinesterase inhibitory activities.the results presented that compounds 4,8,13,14,and 22 showed significant inhibitory activity on acetylcholinesterase,compound 16 exhibited moderate inhibitory activity.Meawhile,compounds 4,8,10,11,13,14,18,22 displayed remarkable inhibitory activity on butyrylcholinesterase,and compounds 7,9,16,19 possessed moderate inhibitory effect.MTT method was used to test the cytotoxicity of some compounds against eight tumor cell lines and one normal cell line.The results showed that compounds 1,3,5,8-14,16-19,21 presented different inhibitory effect on the tumor cell lines.Using a inflammation model of NO production by LPS-induced RAW 264.7 mouse macrophage.The results showed that the treatment of compounds 3,17,28 and 29 can significantly reduce the content of pro-inflammatory factor NO produced in the cell,while compound 30(IC50>100 ?M)showed weak activity.Using a model by H2O2 to induce damage on SH-SY5Y cells.The results showed that after treatment with compounds 28,29,and 30,not only can repair the morphology of nerve cells and cell survival rate significantly improved.In this study,through the separation,identification and simple detection of some activities of the chemical components in Inonotus obliquus,provides scientific basis for the further exploration of the medicinal value of Inonotus obliquus.