Study On The Chemical Constituents Of Anti-influenza Virus Isolated From The Roots Of Baphicacanthus Cusia (Nees) Bremek From Guizhou Province

Nan-ban-lan-gen is the root of Baphicacanthus cusia?Nees?Bremek,which has been used as the medicine and as a material of many TCM formulations for a long history in China.The formulations of Nan-ban-lan-gen usually have good antviral and anti-inflammatory effects.With the frequent outbreaks of inflenza in recent years,B.cusia has gradually become a research hotspots.However,the effective conponents of B.cusia is still unclear.This thesis focuses on study of the chemical constituents of Nan-ban-lan-gen which collected from Dushan,Guizhou province,and their bioloical activities such as anti-influenza virus,antioxidant and anticancer.Finally,the content of six active constituents of different parts of B.cusia in different habitats were analyzed.Aiming to as above,twenty-six compounds were isolated and identified from the extracts of methanol of the roots of B.cusia using various chromatographic and spectroscopy techniques,including eight phenylethanoid glycosides:Acteoside?1?,Isoacteoside?2?,Isocrenatoside?3?,Martynoside?4?,Isomartynoside?5?,2-?3,4-dihydroxy-phenyl?ethyl-?6-O-caffeoyl?-?-D-glucopyranoside?6?,Decaffeoylverbascoside?7?,?-phenylethoxy-?-D-glucopyranoside?8?,four benzoxazinone and compounds:?2R?-2-O-?-D-glucopyranosyl-5-hydroxy-2H-1,4-benzoxazin-3?4H?-one?9?,?2R?-2-O-?-D-glucopyranosyl-2H-1,4-benzoxazin-3?4H?-one?10?,7-Chloro-?2R?-2-O-?-D-glucopyranosyl-2H-1,4-benzoxazin-3?4H?-one?11?,2-hydroxy-?2H?-1,4-benzoxazin-3?4H?-one?12?,and five lignins:?+?-Syringaresinol?13?,???Episyringaresinol?14?,?+?-syringaresinol-O-?-D-glucoside?15?,5',5'-dimethoxy-4'-hydroxy-2,4,6,8-diepoxylignan-4'-O-?-Dglucopyranoside?16?,Elenther osideE?17?,andtwosteroids:Stigmasterol-?-D-glucopyranoside?18?,?-Daucosterol?19?,and two Triterpenoids:Lupeol?20?,Betulin?21?,as well as five other compounds:2-Benzoxazolinone?22?,Iisobutyl phthalate?23?,Dibutyl phthalate?24?,Niacinamide?25?,Squalene?26?.Among them,compound 3?6?7?8?9?11?12?13?14?15?16?17?19?23?24?25?26 were isolated from this plant for the first time.The isolated 20 compounds were screened for anti-influenza virus activity,three of them?3,18 and 19?showed significant activities against influenza?H3N2?virus(IC₅₀:1.54?g/mL-12.83?g/mL).On the other hand,13 compounds were screened for their antioxidant activity?DPPH?,compounds 1,2,3,4,5 and 15 showed strong antioxidant activity(IC₅₀:1.03?g/mL-8.62?g/mL),the strongth order of which was2>1>5>4>15>3 under the concentration circumstance of 1.56?g/ml and 3.125?g/mL,respectively by ORAC method.Anticancer activity of compounds 1,2,3,and 9 were also studied,it was found that Compound 1 had a significant growth inhibitory activity against tumor cell line CB3 in vitro with the IC₅₀=2.26±0.22?M.This thesis also established a RP-HPLC method for analysis of bioactive constituents such as tryptanthrin?1?,2?3H?-benzoxazolinone?2?,indirubin?3?,indigo?4?,acteoside?5?,martynoside?6?,in oder to Analysis of the content of these compounds in different habitats and different parts of B.cusia.The resluts indicated that the content of tryptanthrin?1?was the highest in the upper parts of B.cusia in Luokun County,Guizhou province;the content of indigo?3?was the highest in the upper parts of B.cusia of Guilin City,Guangxi province;B.cusia distributed in Paidao Village,Danzhai County,Guizhou province,the content of aconiside?5?was the highest in the aerial parts;the contents of 2-benzoxazolone?2?in the rhizomes and roots of B.cusia distributed in Honghe of Yunnan province were higher;the contents of indigo?4?in the upper parts of B.cusia of Guilin,Guangxi province were higher than the other places;and the content of flavonoids?6?in the upper part of B.cusia in Xiongqi Village,Danzhai County,Guizhou province is relatively higher,respectively.Many compounds with anti-influenza,anti-oxidation and anti-tumor activities were first identified from Nan-ban-lan-gen distributed in Guizhou Province.Also,a convenient and rational metheod was established for the quality-evaluation and quality-control of Nan-ban-lan-gen.The results of this thesis provide the evidence for the utilization of this medicinal plant,and provide reference for development and research of anti-influenza,anti-oxidation and anti-tumor new drugs from traditional Chinese medicine.