Active Compounds Of Chickpea Against Diabetes Based On DPP-Ⅳ Target

Diabetes mellitus is a disease that glucose,protein and fat metabolism disorder,mainly for high blood glucose and urine glucose.Sustained hyperglycemia can lead to many complications,such as blindness,cardiovascular disease,kidney failure,et al.Its incidence is trending to rise year by year,and it has become one of the major disease that harm to the human health.In recent years,with the study of pathogenesis of diabetes and drug action mechanism,the research of new hypoglycemic drug is also developing rapidly.In numerous antidiabetic,DPP-Ⅳ inhibitors is always the key point of the research.Related studies showed that Xinjiang Cicer arietinum L.has a certain degree of hypoglycemic effetal,at the same time there are a large number of literature reported on chemical constituents and pharmacological effetals of chickpeas.But there are no reports in the literature showing which chickpeas extract have any DPP-Ⅳ inhibitory activity.This research expected that through the establishment of the DPP-Ⅳ screening model in vitro,separation and identify compounds can inhibit DPP-Ⅳ and find out its material basis and mechanism.The assays were conducted to establish a screening model in vitro of DPP-Ⅳ and to investigate the inhibit activity of 30%,50%,70%,95% ethanol elution and N-butyl alcohol,ethyl acetate extract.The results find out that the ethyl acetate extract and 50%,70%,95% ethanol elution have DPP-Ⅳ inhibit activity.The Cicer arietinum L.is separated on a silica gel column chromatography and RP-18-reversed-phase silica gel column.After separation processes by preparative HPLC of active fractions,thirteen compounds were obtained,and their structure were identified as biochanin A(1),biochanin A-7-O-β-D-glucopyranoside(2),formononetin(3),ent-6b,7b,13b-trihydroxy-kauran-19-carboxylic acid-17-O-β-D-glucopyranoside(4),(3S,5R,6S,9S)-3,6-dyhydroxyl-5,6-dihydro-β-ionol(5),(3R,5S,6R,9S)-3,6-dyhydroxyl-5,6-dihydro-β-ionol(6),(3R,5S,6R,9S)-3-acetyl-5,6-dihydro-β-ionol(7),N,N,N-trimethyl-hypaphorine(8),(1S,3S)-1-methyl-1,2,3,4-tetrahydro-β-carboline-3-carboxylic acid(9),7-phenyl-Obtucarbamate(10),N-N’-dioxo formyl-4,4’-diaminodiphenyl methane(11),3-Acetyl indole(12),triethyl citrate(13).Compounds 4,6,7,10 are isolated from the nature for the first time,compounds 5,11,12,13 are isolated from Cicer arietinum L.for the first time.Invistigate the compounds isolated from chickpea extraction such as biochanin A(1),biochanin A-7-O-β-D-glucopyranoside(2),formononetin(3)and genistein can inhibit DPP-Ⅳ.Results showed that the compound biochanin A-7-O-β-D-glucopyranoside(2)exhibiting strong DPP-Ⅳ inhibitory activities in vitro,IC50 = 720 mM.The inhibition of biochanin A-7-O-β-D-glucopyranoside is reported for the first time.This subject researched the chemical composition of chickpeas,and established the DPP-Ⅳ screening model in vitro,provide a scientific basis for the comprehensive development and utilization of Chickpea resources,and has important academic value and application prospect.