Study On The Constituents, Bioactivity And Quality Control Of Resource Plant Maackia Amurensis

Maackia amurensis Rupr.et Maxim.,belongs to the Leaguminosae family,Maackia species, is a deciduous tree distributed widely in the north of China.The dried stem bark of M. amurensis has been used as folk medicine for the treatment of rheumatoid arthritis,pain,allay inflammation,stanch bleeding,etc.In our country,little research on the chemical constituents and pharmacological activities of M.amurensis has been reported.For further development of this medicinal plant material,we conducted research on the chemical constituents,bioactivity and quality control of the stem bark of M.amurensis.To study the chemical constituents of the M.amurensis,the constituents were isolated by various chromatography methods and the structures were elucidated on the basis of chemical and spectroscopic data.54 compounds were isolated from the 70%ethanol extract,including 9 new compounds:maackiaflavanone A（1）,maackiaflavanone B（2）,maackiaisoflavone（3）, maackia- pterocarpan A（4）,maackiapterocarpan B（5）,Maackiaisoflavonoside（15）, maackiachromone（39）,（2S,3S）-7-methoxyflavononol（41）,（-）-maackiacytisine（48）;45 known compounds:abyssinone V（6）,（—）-sigmoidin E（7）,5-hydroxysophoranone（8）, maackiaflavanone（9）,abyssinone II（10）,abyssinone IV（11）,euchrenone b₁（12）,ulexone B （13）,ulexone A（14）,Dalsympathetin（16）,ononin（17）,glycitin（18）,genestin（19）, Saikoisoflavonoside A（20）,Afrormosin-7-O-β-D- glucopyranoside（21）,gehuain（22）, Kushenol O（23）,7-hydroxy-4’,6-dimethoxyisoflavone-7-O-β-D-apiofuranosyl-（1→6）-β-D-glucopyranoside （24）,tectoridin（25）,biochanin A 7-O-β-D- gentiobioside（26）,7-hydroxy4’ -methoxyisoflavone-7-0-β-D-apiofuranosyl-（l→6）-β-.D-glucopy- ranoside（27）,glycitin-6"-O-acetate （28）,ononin-6"-O-acetate（29）,7,5’-dihydroxy-3’-methoxyisoflavone（30）, retusin-8-methylether（31）,7,4’-dihydroxy-3’-methoxyisofiavone（32）,di-O- methyldaidzein （33）,genistein（34）,afromosin（35）,formononetin（36）,prunetin（37）,glycitein（38）,（±）-2, 3-dihydro-3,5-dihydroxy-7- methoxyflavone（40）,3,4-dimethoxy caffeic acid methylester （42）,（-）-maackiain（43）,（-）- medicarpin（44）,ethyl P-hydroxylbenzoate（45）,anisic acid（46）, p-hydroxy benzaldehyde（47）,cytisine（49）,（±）-formylcytisine（50）,N-（3- oxobutyl） cytisine （51）,（-）-epibaptifoline（52）.N-methylcytisine（53）,β-sitosterol（54）.Compounds 6,7.10-14, 16,21-24,26-28,30,31and 45-47 were firstly isolated from Maackia species,and compounds 8,32 were isolated from M.amurensis for the first time.The cytotoxicities against human cancer cell lines（A375S2,HeLa,MCF-7 and HepG2） were tested by the MTT method in vitro,and all the tested compounds,including 4 new compounds:maackiaflavanone A（1）,maackiaflavanone B（2）,maackiaisoflavone（3）, maackiaptecarpan A（4）,demonstrated cytotoxic activities.Among the tested compounds, maackiaflavanone B（2） showed the strongest cytotoxic activity with IC₅0 value of 7.8μM against A375S2 and euchrenone b₁（12） showed the most potent cytotoxicity with IC₅0 value of 4.5μM against HeLa.The kinds of the tested compounds included prenylated flavonones, isoflavones and pterocarpans.We also tested the cytochrome P450 enzyme inhibitory activities of 5-hydroxyl -sophoranone（8）,（-）-maackiain（43）,（-）-medicarpin（44） against CYP1A2、CYP2C9 and CYP3A4.All the three compounds showed inhibitory activities and（-）-medicarpin（44） showed strong inhibitory activity with IC-50 value of 10.7μM against CYP1A2.For quality control of the stem bark of M.amurensis,the content of the total isofavonoids in the stem bark of M.amurensis were determined by UV Spectrophotometry .The HPLC fingerprint was studied on the bark of M.amurensis.Eighteen peaks in the HPLC fingerprint were identified by compared with the standard compounds,including 17 isoflavones.Additionally,the contents of four isoflavanes,genestin,tectoridin, Saikoisoflavonoside A and Maackiaisoflavonoside were determined.These standards will be useful for quality control of the medicinal material.