Studies On Chemical Constituents And Bioactivities Of Rhizomes Of Acorus Calamus L.Crowing In Hunan Province

Acorus calamus L.is the plant of Araceae(Acorus)and mainly distributed in Hu Bei,Hu Nan,Liao Ning,and Si Chuan Provinces in China,it has been commonly used in traditional Chinese medicine to treat phlegm syncope,stroke,epilepsy,palpitation and amnesia,tinnitus and deafness,dyspepsia inducing abdominal pain,dysentery,rheumatic pain,eczema and scabies.Pharmacological studies have shown that it possesses mainly chemical compositions,such as sesquiterpenes,flavonoids,and saponins,and extensive antispasmodic,antimicrobial,antioxidant,antiinflammatory,antidiabetic,nootropic,hypolipidemic,antitumor and insecticidal activities.Because of biological activity and the local rich resources,We continued to systematically research chemical constituents of ethanol extract and volatile oil of A.calamus,and screen biological activities of the crude extract,different polarity parts,separating fragments and chemical constituents,in order to provide a theoretical basis for the rational use and development of local resources.In this thesis,kinds of modern chromatographic and spectral methods were used to isolate the 50 compounds from Rhizomes of A calamus L.,and elucidate their structures.These compounds included 30 sesquiterpenoids(compounds 1-30):1α,5α-guaiane-4α,10α-diol-6-one(1),4β,6β-dihydroxy-1α,5β(H)-guai-9-ene(2),4β,6β-dihydroxy-1α,5β(H)-guai-10(14)-ene(3),teuclatriol(4),calamusenone(5),isocalamusenone(6),spathulenol(7),(-)-alloaromadendrane-3β,9β-diol(8),ent-prelacinan-7-one(9),isocalamendiol(10),calamendiol(11),calamusin H(12),1β,7β(H)-4(15)-cadinadiene-5β,6α,10β-triol(13),1β,7β(H)-3-cadinadiene-6β,10α-diol(14)，T-muurolol(15),1α,7α(H)-cadinane-4β,6β,10β-triol(16),1α,7α(H)-cadinane-4α,6β,10α,15-tetrol(17),cadinadiene A-2,4-epoxide(18),cadinadiene A-1,2-epoxide(19),oxyphyllenodiols A(20),oplodiol(21),ananosmin(22),8-hydroxyl-epishyobunone(23),epishyobunone(24),isoshyobunone(25),Neo-acorane A(26),acoric acid(27),calamusin D(28),(1R,5S,6R,7R,10R)-10-dihydroxy-oplopanane-4-one(29),bullatantriol(30);5 alkaloids(compounds 31-35):tatarine A(31),Neo-tatarine(32),aurantiamide acetate(33),2,6-dioxopiperidin-3-yl acetate(34),palmatine(35);6 benzene derivatives(compounds 36-41):β-asarone(36),a-asarone(37),γ-asarone(38),(Z)-methylisoeugenol(39),2,4,5-trimethoxybenzalde-hyde(40),syringic acid(41);1 sesquinlignans:tatanan C(42);3 sterols(compounds 43-45):cerevisterol(43),daucosterol(44),β-sitosterol(45);4 fatty acids(compounds 46-49):heptadecanoic acid(46),palmitic acid(47),monopentadecanoin(48),monononadecanoin(49);1 monoterpenoids:camphor(50).Compounds 1,13,14,16,17,18,19,23,26 and 29 were new sesquiterpenoids.Compounds 32 was new alkaloid.13 compounds were repoted for the first isolation from the plants of Acorus.14 compounds were isolated from the plant for the first time.This data provided a scientific basis for material basis research of Acorus calamus L.The antimicrobial activities were screened for volatile oil,ethanol extract and different polarity extract parts from Rhizomes of A calamus L.in vitro.The experimental results showed that the volatile oil,ethanol extract and extract parts had more significantly antibacterial activity against gram-positive bacteria,Staphylococcus aureus,S.epidermidis,and Strptococous hemolyticus than gram-negative bacteria,Escherichia coli and Shigella dysenteriae,and aslo had strong inhibition effect agaist Helicobacter pylori and Monilia albicans,in addition to Pseudomonas aeruginosa.Ethanol extract,petroleum ether extract,and ethyl acetate extract had significant inhibitory against,.hemolyticus(MIC 3.13 mg/mL),chloroform extract had significant inhibitory against(MIC 3.13 mg/mL).Compounds 4,22,27,30 and 34 isolated from ethanol extract had significantly inhibitory activity against S.aureus,E.coli,M.albicans,and P.aeruginosa in vitro,in which had good bactericidal effect for compound 22 against above four tested strains(MBC 20.00 μg/mL),compound 30 against P.aeruginosa(MBC 16.62μg/mL),compound 34 against S.aureus(MBC 10.44μg/mL).Directly bioautography was applied to screen inhibition activity of plant fungals of the volatile oil,separating fragments and isolated compounds.Experiment results show that compound 5,6,36,37 and 39,which were isolated from the active fragment and the volatile oil had antimicrobial activities against Colletotrichum acutatum、C.fragariae,and C.gloeosporioides.There were more than 40%inhibition rate for compound 6(150 μM and 300μM),compound 37(150 μM and 300μM),and compound 39(300 μM)against Botrytis cinerea(48 h),compound 37(300 μM)against Fusarium oxysporum(48 h),compound 37(300μM)against P.obscurans(120 h).Herbicidal active experiment showed that the volatile oil and seven compounds isolated from rhizoma Acori calami had no inhibitory effects to the growth of Lettuce.The volatile oil,compound 36(1000μM),compound 37(1000μM)and compound 39(333μM and 1000μW)had significantly inhibited the growth of Agrostis.Compounds 10,11,27,31 and 32 exhibited protective activities on the Aβ25-35-induced injuries of PC12 cells at 30μM;when compound 27(5.0μg/mL),31(0.5 and 1.0μg/mL),and 32(1.0μg/mL)effected the damaged cells,the viability was more than 95%.This data provided experiment basis for clarifying effect of phlegm syncope,stroke,epilepsy and antimicrobial activity.