Studies On The Chemical Constituents And Bioactivities Of Inonotus Obliquus

The medicinal fungus Inonotus obliquus(Pers.:Fr.)Pilat,which belongs to the family Hymenochaetaceae,is widely distributed in Europe,Asia,and North America.In Russia and western Siberia,this mushroom has been used in folk medicine for cancer treatment for more than four centuries.Recently,the extract of this fungus was reported to possess antitumor,antioxidant,and anti-inflammatory activities.During an ongoing search for new bioactive metabolites from fungi,the EtOAc fraction(IO-3)of the 95%ethanol extract exhibited activity in protecting the injury by hydrogen peroxide oxidation on HUVEC.The water insolutable fraction of the 95%ethanol extract(IO-7),the 50%ethanol(10-11)and 70%ethanol(IO-12)fractions eluted from the macroporous resin column of the water extract showed inhibitory activities against PTKs.All fractions of the 95%ethanol extract(IO-2 to IO-7)exhibited selective cytotoxicities against human cancer cell lines.Moreover,fractions 10-11 and 10-12 also showed inhibitory activities against neuraminidase of the influenza vitus.The 95%ethanol extract and water extract of the sclerotia of I.obliquus were investigated respectively on their chemical constituents systematically by various kinds of chromatographic methods.Seventy-one compounds were isolated from I.obliquus,including fourteen new compounds,and their structures were elucidated on the basis of spectroscopic analysis,including homonuclear and heteronuclear correlation NMR('H-'H COSY,HSQC,HMBC,NOESY and ROESY)experiments,in addition to MS,IR,UV and CD experiments.Among the known compounds,fouty-seven compounds were isolated from I.obliquus for the first time.Forty-five compounds were isolated and identified from the 95%ethanol extract of the sclerotia of I.obliquus,including eighteen triterpenes,eight sterols,fourteen aromatic compounds and five fitty compounds.These compounds are listed as follows:inonotusol A(1*),inonotusol B(2*),inonotusol C(3*),inonotusol D(4*),inonotusol E(5*),inonotusol F(6*),inonotusol H(7*),inonotusic acid(8*),inotodiol(9),lanosterol(10),3?,22-dihydroxy-lanosta-7,9(11),24-triene(11),3?,22-dihydroxylanosta-8,24-diene-11-one(12),3?,22-dihydroxylanosta-8,24-diene-7-one(13),trametenolic acid(14),3?-hydroxy-lanosta-8,24-diene-21-al(15),botulin(16),21-hydroxylanosterol(17),3?-hydroxylanosta-7,9(11),24-trien-21-oic acid(18),5,8-epidioxyergosta-6,22-dien-3?-ol(19),ergostol(20),lawsaritol(21),ergosta-7-en-3?-ol(22),ergosta-4,6,8(14),22-tetraen-3-one(23),ergosta-7,22-dien-3?-ol(24),ergosta-7,22-dien-3-one(25),3?,5a-dihydroxy-(22E,24R)-ergosta-7,22-dien-6-one(26),phaeoporlactone(27*),vanillic acid(28),4-acetocatechol(29),protocatechuic aldehyde(30),2,3-dihydroxy-1-(4-hydroxy-3-methoxyphenyl)-propan-1-one(31),2,3-dihydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-1-propanone(32),protocatechuic acid(33),2-methoxyl-benzoic acid(34),1,2-dihydro-2-oxo-4-quinoline-carboxylic acid(35),gallic acid(36),syringic acid(37),(E)-4-(3,4-dihydeoxyphenyl)-but-3-en-2-one(38),benzoic acid(39),phelligridins A(40),2,7-dihydroxy-octanedioic acid(41),palmitic acid(42),monoolein(43),monopalmitin(44),hemisceramide(45).Compounds 1*-8*and 27*are new compounds.Compounds 16,19-26 and 28-45 were isolated from I.obliquus for the first time.Twenty-nine compounds were obtained and identified from the water extract of the sclerotia of I.obliquus,including ninteen aromatic compounds,two nucleosides,five sugers or suger alcohols,one amino acid,one degraded carotenoid and one alkaloid.These compounds are listed as follows:protocatechuic aldehyde(30),(E)-4-(3,4-dihydeoxyphenyl)-but-3-en-2-one(38),phelligridins A(40),inonotusin A(46*),inonotusin B(47*),inonotusin C(48*),inonotusane A(49*),inonotusane B(50*),sinapaldehyde(51),ethyl protacatechuate(52),syringaldehyde(53),caffeic acid(54),trans-cinnamic acid(55),glucosyringic acid(56),aquilarin A(57),3(5)-pryazole-carboxylic acid(58),5-benzofurancarboxylic acid(59),phelligridins D(60),3-methoxy-4-hydroxy-trans-cinamaldehyde(61),adenosine(62),guanosine(63),?-D-glucose(64),a-D-glucose(65),sucrose(66),galactitol(67),mannitol(68),L-phenylalanine(69),glycoric acid(70),betaine(71).Compounds 46*-50*are new compounds.Compounds 51-59 and 61-71 were isolated from I.obliquus for the first time.Among the compounds isolated from the 95%ethanol extract,nine compounds(1*,2*,12,13,14,27*,28,30 and 31)exhibited activities in protecting the injury by hydrogen peroxide oxidation on HUVEC(10 ?g/ml).Three compounds(27*,29 and 30)showed inhibitory activities against PTKs,with IC50 values of 24.9,24.6 and 7.7 ?mol/L,respectively(10 ?g/ml).Eight compounds(9,27*,29,30,38,40,47*and 60)exhibited selective cytotoxicities against KB(human nasopharynx cancer cell line),Bel-7402(human hepatoma cell line),A-549(human lung epithelial cell line),BGC-823(human gastric cell line),or A2780(human lysate cell line)cell lines by MTT method,with IC50 values of<10 ?mol/L or 5 ?g/mL,respectively,and using 5-fluorouracil or taxol as the positive control.Morover,the compounds isolated from the 95%ethanol extract were also bioassayed for their hepatoprotective effects against D-galactosamine-induced toxicity in WB-F344 cells,using the hepatoprotective activity drug bicyclol as the positive control.Ten compounds(6*,8*,9,10,12,14,21,22,24 and 28)showed hepatoprotective activities(10 ?mol/L).