Studies On The Chemical Constituents Of Veratrum Nigrum L., Veratrum Japonicum (Baker) Loes.F. And Leonurus Heterophyllus Sweet, And Studies On Mechanism Of Processing Of Veratrum Nigrum L.

The large genus Veratrum is belong to Liliaceae family. The dried roots and rhizomes of Veratrum Linn. is a extremely poisonous chinese herbal medicine, possessing the functions of hypotensive activity, emetic, antithrom and antifungus. At the same time it also has toxicities of irritant action on digestive tract mucosa, nucleus nervi vagi and central nervous system.According to theory of processing chinese herbal medicine, we processed the dried roots and rhizomes of Veratrum nigrum L. with rice vinegar. The systematic investigation on chemical constituents led to the isolation of fourteen compounds. On the basis of chemical evidences and spectral analysis, the structures of the 14 compounds were elucidated, and they are 12β-hydroxylveratroylzygadenine(1)^*, 1β, 3α-dihydroxy-5β-jervanin-12-en-11-one (2)^*, neoverapatuline(3)^*, veratramine(4), veratrosine(5), rubijervine(6), gadenine (7), jervine(8), veramarine(9), verdine(10), pseudojervine(11), resveratol(12), piceid(13),β-sitosterol(14). Among them, 1, 2 and 3 are new compounds.The systematic investigation on chemical constituents of crude alkaloids of the dried roots and rhizomes of [Veratrum japonicum(Baker)Loes. f. ] led to the isolation of 6 compounds. On the basis of chemical evidences and spectral analysis, the structures of the 5 compounds were elucidated, and they are neogermine(15)^*, germine(16), germerine(17), neogermbudine(18), veramitaline(19). Among them, compound 15 is new compound.The quantitative analysis of veratramine (9) from crude drug of Veratrum nigrum L. and drag processed with rice vinegar of Veratrum nigrum L. was performed by HPLC; The quantitative analysis of veratrosine(5)from crude drug of Veratrum nigrum L., drug processed with rice vinegar of Veratrum nigrum L. and drug processed with mixture of rice flour and water of Veratrum nigrum L. An HPLC method was developed for the determination of resveratol(12) and piceid(13) in crude drug of Veratrum nigrum L. and drug processed with rice vinegar of Veratrum nigrum L.. An HPLC method was developed for the determination of 10 compounds in crude drug of Veratrum nigrum L., drug processed with rice vinegar of Veratrum nigrum L. and drug processed with mixture of rice flour and water of Veratrum nigrum L..Some of these compounds were tested for their antitumor activity (cancer cell HL-60 and SF188). 12β-hydroxylveratroylzygadenine (1), veramarine (9) and veratroylzygadenine (7) showed moderate cytotoxicity against HL-60 cell lines, and veratramine (4) and rubijervine (6)exhibited weak cytotoxicity against SF188 cell line.Evaluation of acute toxicity on water extracts of crude drug of Veratrum nigrum L. and of drug processed with rice vinegar of Veratrum nigrum L. showed LD₅₀ of water extracts of crude drug of Veratrum nigrum L. on mice by lavage is 4.2g/kg with 95% confidence level of 3.4～5.4g/kg; LD₅₀ of water extracts of drug processed with rice vinegar of Veratrum nigrum L. on mice by lavage is 3.2g/kg with 95% confidence level of 2.5～3.9g/kg.Measuring the effect of the water extract of Vertrum japonicum (Baker)Loes. f. on locomotor activity in mice showed that the water extract of Vertrum japonicum (Baker)Loes. f. have the function of central inhibition in extent of 0.5-1.40g/kg with ED₅₀ 0.684 g/kg.We investigated in vivo the DNA damage induced by water extract, crude alkaloid, neogermine(15), germine(16), germerine(17) and neogermbudine(18) from the dried roots and rhizomes of Veratrum japonicum(Baker) Loes. f. in mouse brain cells by using single cell gel electrophoresis.Leonurus heterophyllus Sweet is a grass widely distributed in the Eurasia, which belong to Labiatae family. It possesses the functions of promoting blood circulation to remove blood stasis, detumescense and diuresis. It is available in curing irregular menstruation, puerperal congestion, cardiocerebrovascular diseases, blood diseases and so on.From the water extract, 95% ethanol extract and 70% ethanol extract of the aerial part of Leonurus heterophyllus Sweet, by solvent extraction, chromatograghic and recrystalllized methods, 14 compounds were isolated. On the basis of their physicochemical properties and spectroscopic analysis, their structures were identified as heteronoside(20)^*, apigenin (21), quercetin(22) tiliroside (23), rutine(24), quercetin-3-O-robinobioside(25), megstigmane (26), leonurine (27), ajugoside (25), 2, 6-dimethyl-2E, 7-octadiene-1, 6-diol (29), syringic acid (30), benzoic acid (31),β-sitosterol(32), daucosterol(33). Among them, compound 20 is new compound and compounds 25, 26, 29, 30, 31 were isolated from this genus for the first time.