Study On The Constituents And Bioactivities Of Medicinal Fungus Fomes Officinalis And Fomes Fomentarius

Fomes officinalis, a fungus of Fomes genus in Polyporaceace family, is a wood rotting fungus which is found on the trunks of living or dead coniferous trees in the northern regions of China and in the Pacific Northwest United States, Canada and in Europe. It is traditionally used in Chinese Uigur prescription to treat cough and asthma. As a folk medicine, the fruit body of F. officinialis is traditionally used in the treatment of coughs, gastric cancer, rheumatism and hydropsia. Modern pharmacological studies have shown that the extract of A-Li-Hong have a variety of pharmacological action including:anti-tumor, eliminating phlegm, scavenging hydroxyl radical and enhancement of anti-stress. However, there have been no quantitative method to control the quality of crude drug in the Drug Specifications Promulgated by the Ministry of Public Health, PR China. Therefore, a simple and accurate HPLC-DAD-ESI/MS method was developed in this study for the simultaneous quantification and identification of the main bioactive constituents in F. officinalis. In order to obtain a more comprehensive understanding of its effective physiological constituents, in this doctoral dissertation, the systematic phytochemical and pharmacological studies have been carried out.Thirty-four compounds were isolated from the ethanol extracts of its dried sclerotium. On the basis of various modern spectroscopic analysis(IR, UV, MS,'HNMR,13CNMR, 'H-'HCOSY, HMBC, HSQC, DEPT) and classical chemical methods, thirty-twor compounds were determined, of which three were new, seven were isolated from this fungus for the first time. The new compounds'structures were determined as follows:fomic acid(3, 7-dihydroxy-driman-l 1-oic acid), fomefficinin(18,23-epoxy-9a-ethoxy-3a-carbohydroxy acetyloxy-24a-methyl-lanost-8-en-26,23-olide),2-(1-carboxyhexadecylamino)-2-amino-succinic acid.The known compounds were identified as albicanic acid, fomitopsins C, eburicol,3-keto-dehydrosulfurenic acid, dehydroeburicoic acid, eburicoic acid, sulphurenic acid, dehydrosulphurenic acid, dehydroeburiconic acid, versisponic acid D, versisponic acid C, fomefficinic acid A, fomefficinic acid B, fomefficinic acid C, fomefficinic acid D, fomefficinic acid E, fomefficinic acid F, fomefficinic acid G, fomefficinol A, fomefficinol B, officinalic acid, fomlactone B, fomlactone A, fomlactone C, laricinolic acid, ergosterol, ergosta-7,22,dien-3(3-ol, ergosta-5,22,dien-3β-ol, dodecane.A simple and accurate HPLC-UV method was developed for the simultaneous quantification of bioactive triterpenes in the fruit bodies of F. officinalis. The developed assay could be considered as a suitable quality control method for this folk medicine. Fragmentation study of isolated lanostane type triterpenes in F. officinalis by LC-ESI could help to identify the constituents with skeleton of lanostane triterpene.We tested 12 compounds by acetylcholinesterase activity model, with the minimum and maximum concentration of 5μg/mL and 50μg/mL. The results show that major of the comounds performed weak activity compared with the positive control and they showed inhibition activity with a concentration-dependent.At the same time,Some of the compounds were tested for cytotoxicity against six human cancer lines in vitro.We found that the new sesquiterpene named fomic acid showed inhibitory activity to HL-60 and BGC-823 cell lines at a high concentration, otherwise, the other compounds do not have obvious activity but concentration-dependent.HL-60, Bel-7402 and KB cancer lines in vitro, and they appeared to be modestly active.Fomes fomentarius(L.Ex.Fr.), is a folk medicinal fungus of Polyporaceae in family. It has the function of anti-tumor, eliminating phlegm, scavenging hydroxyl radical and enhancement of anti-stress. In order to find its effective compounds, we made a systematic research on this fungus.Using kinds of chromatography technique,16 compounds were isolated from the seeds alcohol extract of F. fomentarius. On the basis of various modern spectroscopic analyzing,14 compounds were determined, The new compound's structure was determined as:fomentarinin (ethyl tetrahydro-5-tridecylfuran-2-carboxylate) (Ⅰ). The known compounds were identified as paulownin (II), protocatechualdehyde (Ⅲ), 4-(3,4-dihydroxyphenyl)-but-3-en-2-one (IV), syringic acid (V), syringyl alcohol(VI), vanillin(VII),5α,8a-epidioxyergosta-6,22-dien-3β-ol(VIII), ergosta-7,22-dien-3β-ol (IX), ergosta-7,22-diene-3,6-dione (X), ergosta-7,22-dien-3-one(XI), ergosterol(XII), P-sitosterol (XIII),2-hydroxy-hexacosanoic acid ethyl ester(XIV). Compound II-Ⅷ,X,XIII and XIV were isolated from F. fomentarius for the first time.Some of the compounds were tested for cytotoxicity against four human cancer lines in vitro. We found that protocatechualdehyde and 4-(3,4-dihydroxyphenyl)-but-3-en-2-one showed inhibitory activity to BGC-823 and Bel-7402 cell lines at a high concentration.