Study On Diterpenoid Alkaloids From The Whole Plant Of Aconitum Tanguticum

Aconitum tanguticum is the whole plant of Aconitum tanguticum(Maxim.) Stapf, and it is a commonly used traditional Tibetan medicine and has a tibetan name Ponka. Aconitum naviculare(Bruhl.) Stapf is also another plant of Ponka, Ponka is firstly recorded in Somaratsa, It is cool in nature, bitter in taste and slightly toxic, and has the effects of reducing fever and removing toxicity. Ponka is usually used in the Tibetan patent medicines for treating infectious fever, influenza, hepatitis, cholecystitis, pneumonia and food poisoning etc. Modern phytochemical study results showed A. tanguticum contanined various kinds of chemical constituents, inclucing diterpenoid alkaloids, flavonoids,phenolic acids and volatile oils. Pharmacological studies showed that A. tanguticum has antiviral, antimicrobic, antiinflammatory, antitumor and antiarrhythmic effects.In the study, 43 compounds were isolated from A.tanguticum total alkaloids fraciton by using various column chromatographis and chemical methods, such as silica gel, ODS,Sephadex LH-20, preparative HPLC, preparative TLC and recrystallization and so on. The structures of 30 compounds were elucidated based on physical and chemical property analysis, spectroscopy technology(UV, MS, 1H-NMR, 13C-NMR, HSQC, HMBC and1 H,1H-COSY), x-ray analysis and comparison with the literature data. Among them, 20 compounds are alkaloidss(including 9 diterpenoid alkaloids):They are 2β,9β,11β,13β-tetra-hydrohetisine, hetisane-2α,11α,13α,19β-tetrol-2-benzoate-11-sulfonate, 13-dehydro-2β,3α,6β-trihydroxyhetisine, sinapic amido-4-O-β-D-glucopyranoside, 11α,13α-dihydroxyhetisin-one, heteratisine,tanaconitine, guan-fu base H, benzamide, hetisine, N-oxide hordenine,candicinechloride, dihydroatisine, isolumichrome, p-hydroxy phenylethylamine,3-(4-aminophenyl) propenoic acid, adenosine, 3-(p-hydroxyphenyl) propionic acid amide,N-methyltyramine, cyclo-(Ile-Leu), phenylalanine. 5 phenolic acids, They are p-hydroxy phenylethanol, p-hydroxycinnamic acid, butylparaben, salidroside, trans-ferulic acid-4-O-β-D-allopyranoside) and 4 other compounds, They are β-sitosterol, phthalic acid butyl isobutyl ester, benzoic acid,(2R,3R)-2,3,4-trihydroxy-butanoic acid). Compound 1~4 are new compounds, 24 compounds are isolated from this species for the first time. The antiviral activity of 7 diterpenoid alkaloids(11α,13α-dihydroxyhetisinone, heteratisine,tanaconitine, guan-fu base H, hetisine, 2β,9β,11β,13β-tetrahydrohetisine,13-dehydro-2β,3α,6β-trihydroxyhetisine)were tested by CPE method, and the result showed 11α,13α-dihydro-xyhetisinone has good inhibition effect on the growth of H1N1 virus.In order to efficiently control the quality of Ponka, its quality standard was established in this study. Ponka is recorded in Chinese Pharmacopeia(2015 edition), Which is ranunculaceae plants of the genus Aconitum(Aconitum tanguticum(Maxim.) Stapf) or(Aconitum naviculare(Bruhl.) Stapf) dry grass. The tests of water content, total ash and ethanol-soluble extractives of the crude drug were carried out based on the methods recorded in appendix of Chinese Pharmacopeia(2015 edition, volume 4). The content of total alkaloids in Ponka was derermined by ultraviolet spectrophotometry, using atisine hydrochloride as the reference substance, The content of total flavonoid glycosides was assayed by HPLC on a Welch C18 column, using quercetin and kaempferol as the reference substances. The moisture content, total ash content, total ethanol-soluble extractives, total alkaloids and total flavonoid glycosides of 17 batches samples were varied in the ranges of7.23%~10.2%, 5.94%~18.6%, 11.3%~29.4%, 0.386%~1.99% and 0.420%~3.09%, the average content were 8.31%, 10.7%, 21.4%, 0.896% and 1.63%, respectively. According to the limitation standards establishment principle in 2015 edition of Chinese Pharmacopoeia,this paper tentatively recommended the moisture content should not be over 12.0%, the total ash content should not be over 15.0%, the ethanol-soluble extract mass fraction should not be less than 17.0%, the total alkaloids content should not be less than 0.4% and total falvonoid glycosides content should not be less than 0.8% for Ponka.This paper also summarized the research progresses of Tibetan medicines of Aconitum genus, including the research on the genus judgement and geographic distribution, chemical constituents, pharmacological activities, historical evolution of processing methods and toxicities of these Tibetan medicines, and the review will provide theoretical basis for the use of Tibetan medicinal plants of Aconitum genus.