Study On The Active Constituents From Solanum Indicum And Solanum Lyratum For Anti-H5N1Influenza Virus

Influenza A viruses is one of the most important biological threats, not only forpoultry but also for other avian species and humans. Although human infection hasbeen rare, the highly pathogenic avian influenza virus (HPAIV) of the H5N1oftencauses a lethal illness, and reports of their sporadic human-to-human transmissionhave raised concern over the potential for a deadly new influenza pandemic. Screeningsafe and effective anti-HPAI compounds in Chinese herbal medicines provides a largeopportunity for the development of anti-HPAI drugs. In the dissertation, we selectedA/Tiger/Harbin/01/2002(HAB/01) activity screening model for H5N1virus strains toinfect MDCK cell and chose cytopathiceffect (CPE) as activity-evaluating method. Twosolanum species, solanum indicum and solanum lyratum, showed prominent anti-HPAIactivity. So our studies on chemical constituents and anti-HPAI activity of S. indicumand S. lyratum possess a great significance.S. indicum, from the family Solanum, mainly distributed in Yunnan province. Inthe research of the active constituents from the plant, the ethanol extracts of the plantwere systematically chemical investigated, getting sixty-seven compounds bychromatography methods. And the structures of sixty compounds were elucidated onthe basis of physical and chemical evidences and modern spectral analysis (includingIR，UV，1H-NMR，13C-NMR，HSQC，HMBC，MS，et al.). The structures of sixtycompounds were as follows: indicumine F(1), indicumine G(2), indicumine H(3),indicumine I(4), indicumine J(5),7-hydroxy-β-carboline-1-methylpropionate(6),10-oxo-N-trans-feruloyl tryptamine(7),10-oxo-N-trans-feruloyl-5-hydroxy-tryptami-ne(8), indicumine A(9), indicumine B(10), indicumine C(11), indicumine D(12),indicumine E(13),7-hydroxy-6,8-dimethoxy-3-(4’-hydroxy-3’-methoxyphenyl)coum-arin (14),7-hydroxy-1-carbonyl-1,2,3,4-hydrogen-carboline(15), indicumine K1(16),indicumine K2(17), N-trans-feruloyltyramine (18) tribulusamide A(19), flavumindole(3a)(20),7-hydroxy-1-(4-hydroxy-3-methoxyphenyl)-N2,N3-bis(4-hydroxyphenethyl)- 6-methoxy-1,2-dihydronaphthalene-2,3-dicarboxamide (21), thoreliamide C(22),erythro-canabisine H(23) hyoscyamide(24), grossamide K(25), N-trans-feruloyltryptamine(26), moschamine(27), arenarine D(28),7-hydroxy-3,4-dihydro-1-oxo-β-carboline(29), aervin(30),3-indole aldehyde(31),3-indole acid(32),3-hydroxy-2-phenyl acrylic amide(33), isofraxidin(34), scopoletin(35), fraxinol(36),5,7-dihydroxy-6-methoxy coumarin(37), arteminorin A(38), arteminorin B(39),4,4’-biisofraxidin(40), cleomiscosin A(41), medioresinol(42), vanillic acid(43),vanillin(44), ferulic acid ethyl ester(45), genistein(46), genstin(47), rubimaillin(48),kaempferol(49),6"-O-Acetylgenistin(50), munjistine(51), α-D-galactose ethylester(52),1,3–dihydroxyanthraquinone(53), C16acid(54), daturaolone(55),daturadiol(56), hyoscyamilactol(57), larnaxolide A(58), β-sitosterol (59),β-daucosterol (60). Compounds1-17were new compounds, including nine amides,indoles, six coumarins, and two lignans. Compounds20,22~30,33, and38~41, wereisolated from the genus Solanum for the first time. Compounds19,21,31~32,35~37,and42~60were isolated from Solanum indicum for the first time. S. lyratum is a plantof the Solanum family and distributes in the southern provinces of China. We alsoisolated eighteen compounds by chromatography methods from this plant, Theirstructures were elucidated as: lyratoside A(1), lyratoside B(2), lyratoside C(3),lyratoside D(4), lyratoside E(5), lyratoside F(6), lyratoside G(7),(25R)-26-O-β-D-glucopyranosyl)oxy]-22α-methoxy-5α-furostan-3β-O-β-D-glucopyranosyl-(1â†'2)-O-β-D-glucopyranosyl-(1â†'4)-β-D-galactopyranoside(8),26-O-β-D-glucopyranosyl-(22,25R)-3β,26-dihydroxy-22-methoxy-furost-5-ene-3-O-α-L-rhamnopyranosyl-(1â†'2)-β-D-glucopyranosiduronic acid methyl ester(9), pallidifoside B(10),soladulcidine(11), cholesteryl(12),5α-stigmanstan-3,6-dione(13),4-(2,6-dimethylbenzyl)-5,5-dimethyl-dihydro-2(3H)-furanone(14). Compounds1-7were newcompounds, including two steroidal alkaloids and five steroid saponins, Compounds8~13, and14were isolated from Solanum lyratum L. for the first time.A series of compounds, including10amides,14coumarins and15indoles, wereisolated from the seeds of Solanum indicum and enriched the chemical compositionsof this plant. Indicumine F(1), indicumine G(2), and indicumine H(3) are novel skeleton compounds. More than five new compounds, which were isolated from S.lyratum, have3β-O-β-D-glucopyranosyl-(1â†'2)-O-β-D-glucopyranosyl-(1â†'4)-β-D-galactopyranoside connecting to C-3position. Lyratoside D is a new nucleus from thegenus for the first time.At the effective dose of1.2μg/mL,2.4μg/mL,18.75μg/mL,37.5μg/mL,150μg/mL, the compounds X, Y, munjistine, kaempferol,3-indole acid didn’t exhibitapparent harming effects on cell viability, metabolism or proliferation, possessing apotent anti-influenza virus activity in MDCK cell cultures infected with H5N1subtype. Compared with the control group, compound X could extend the survivaltime of BABL/c mice for1~2days in vivo. Compound X might become leadingnatural compounds for the systhesis and development of new anti-HPAI drugs.The result of the research was meaningful to a widespread usage of these towplants, it is also helpful for the development of a new medicine.