| Bauhinia championii(Benth.) Benth., which comes from the bauhinia plant of caesalpinioideae of leguminous, is one of the traditional chinese medicines for treating rheumatoid arthritis, pain and cardiovascular disease.In this paper, 5 polymethoxyflavones(PMFs) from Bauhinia championii(Benth.)Benth. were isolated by the silica gel column chromatography, sephadex LH-20 column chromatography and semi-preparative reversed phase liquid chromatography, and were purified by recrystallization. The structures of PMFs were identified as 5,6,7,3’-tetramethoxy-4’,5’-methylenedioxyflavone(PMF1), 5,6,7,3’,4’,5’-hexamethoxyflavone(P MF2), 5,6,7,3’,4’-pentamethoxyflavone(PMF3), 5,7,3’-trimethoxy-4’,5’-methylenedioxyflavone(PMF4) and 5,7,3’,4’,5’-Pentamethoxyflavone(PMF5) using 1H and 13 C NMR in combination with literature and physicochemical characters analysis.TLC identification method of Bauhinia championii(Benth.) Benth.,which was used PMFs as reference substances, chosen petroleum ether(30-60 ℃)-water saturated ethyl acetate(3:10) for double outspread solvent and detected at 365 nm in the UV lamp, was successfully established for supportting to set up quality control standard of Bauhinia championii(Benth.) Benth..A validated DAD-HPLC method was developed for simultaneous quantification analysis of 5 PMFs of total flavones from Bauhinia championii(Benth.) Benth..C18 column, diode array detector and a mobile phase of acetonitrile-water(35:100, v:v) at a flow rate of 1m L·min-1 in isocratic elution mode have been used in HPLCanalysis. Besides, PMF1 and PMF3 were detected under 332 nm while PMF2, PMF4,PMF5 were acquired datas from 320 nm, 338 nm, 328 nm respectively. The developed method was fully validated in terms of linearity, accuracy, precision and system suitability and were successfully applied to simultaneous quantify the investigated compounds. The result showed that total flavones was containing PMF1(15.92%,w/w), P MF2(26.67%,w/w), PMF3(9.11%,w/w), PMF4(5.12%,w/w) and PMF5(23.35%,w/w).The average total amount of PMFs of total flavones was 80.18%.During the past decades, a number of studies have demostrated multiple biological effects of Bauhinia championii(Benth.) Benth. and total flavones from Bauhinia championii(Benth.) Benth.. In this study, on one hand, it was aimed to forecast targets related with rheumatoid arthritis(RA) and cardiovascular disease(CVD), which linked closely with the main effects of Bauhinia championii(Benth.) Benth., throught the targets of PMFs. On the other hand, it was expected to excavate potential pharmacological activities of PMFs and PMFs riching fractions, such as total flavones, through analyzing targets of PMFs as well. Targets of PMFs were evaluated and rapidly forecasted with Pharm Mapper, a reverse pharmacophore-based screening platform,and as well as with drug target databases, including PDTD, Drug Bank and TTD. Targets were comprehensively analyzed through pharmacology approach. In the end, a total of 106 homo sapiens targets were identified for five PMFs. Five targets with anti-RA potential including DHODH, BACE1, MMP8, MAPK14, CTSK and sixteen targets related with CAV including CDK2, BACE1, CCNA2, ESR2, MME, F2, MA PK10, FGFR1, HMGCR, SORD, MAPK14, MMP12, MMP3, PDE5 A, PLA2G2 A and SELE, were screened, respectively. Moreover, when the targets were classified into groups according to their related diseases, cancer was in the spotlight with 38 targets and there were 9 targets related with diabetes. Overall, this research provided a strong clue for the study of pharmacological activities and mechanisms of action of PMFs and PMFs-rich substances by analyzing the corresponding “drug-target-dise ase” interaction network. |
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