Studies On The Chemical Constituents And Biological Activities Of Ilex Hainanensis And Miliusa Balansae

Ilex asprella.（Aquifoliaceae）is mainly distributed in the south of the People’s Republic of China.The roots of Ilex asprella has been used as Traditional Chinese Medicine for treating secularism,headache,vertigo,diarrhea,laryngalgia,gonorrhea, wound and so on.There are many reports on clinical research of "gang-mei" for treatment of cold.However,there were few reports on chemical constituents and biological activities of this plant.In order to expand the use of "gang-mei",to explain the chemical base of biological activity,and to found out the quality control,the chemical research of Ilex asprella was carried on.The biological activities of some compounds were also studied.Miliusa balaecae（Annonaceae）is widely distributed through out Guangdong,Guangxi,Guizhou,Yunnan,and Hainan provinces.It is used to treat rheumatism,gastropathy and glomerulonephropathy as herb medicine.In order to find novel lead-compound with bioactivity,the chemical research of Miliusa balaecae was carried on.The biological activities of some compounds were also studied.The contents of this paper are as follows:1)Chemical research of Ilex asprella:chromatography and spectrum methods were employed for isolation and identification of the chemical constituents.25 compounds were isolated and identified from the 50%and 95%ethanol extract of Ilex asprella.Among them,11 were triterpenoid saponins.Their structures were identified as follows:Ilexasprellanoside A（1）,Ilexasprellanoside B（2）,Ilexasprellanoside C（3）, Ilexasprellanoside D（4）,Ilexasprellanoside E（5）,Ilexasprellanoside F（6）, Ilexasprellanoside G（7）,3β-[（α-L-arabinopyranosyl）-oxy]urs-12,18-dien-28-oic acid（8）,suavissimosideR1（9）,3β-[（α-L-arabinopyranosyl）-oxyl]-19α-hydroxyurans12-en-28-oic acid 28-β-D-gluco-pyranosyl ester（10）,oleanolic-acid-3-O-β-glucurono- pyranoside（11）.4 were triterpenoids.Their structures were identified as follows:3β-acetoxy-28hydroxyurs-12-ene（12）, pomolic acid（13）,2β,3α-dihydroxyur-ursolic-acid（14）, oleanolic acid（19）.10 belonged to other types.Their structures were identified as follows:3-O-[6’-O-palmitoyl-β-D-glucosyl]-stigmasta-5, 25（27）-diene（15）,3-O-[6’-O-stearoyl-β-Dglucosyl]-stigmasta-5, 25（27）-diene（16）,3-O-β-D-glucosylstig-masta-5,25（27）-diene （17）,syringic acid（18）,palmitate（20）,octacontane（21）,hentriacontane（22）, 1-dotriacontanol（23）,glucose（24）,sitosterol（25）.Among these compounds,compounds 1,2,3,4,5,6,and 7 were new compounds. All the known compounds were isolated from Ilex hainanensis for the first time except for 17,19.2)Chemical research of Miliusa balansae Fin.et Gag..Chromatography and spectrum methods were employed for isolation and identification of the chemical constituents.23 compounds were isolated and identified from the 80%ethanol extract of Miliusa balansae.8 were phenolic glycosides,Their structures were identified as follows: miliusoside A（26）,miliusoside B（27）,miliusoside C（28）,2-（4-hydroxylphenyl） ethyl-O-β-D-apiosyl-O-（1â†'6）-β-D-glucopyranoside（29）,alang ionoside B（30）, cuchiloside（31）,1-（α-L-rhamnosyl-（1-6）-β-D-glucopyranosy-loxy）-3,4,5-trimethoxybenzene（32）, 3,4,5-trimethoxyphenol-β-D-glucopyran-oside（33）.6 were alkaloids.Their structures were identified as follows:norushinsunine（34）, allantion（35）,（+）-coclaurine（36）,（-）-N-methyl-coclaurine（37）,liriodenine（39）, reticuline N-oxide（41）.9 belonged to other types.Their structures were identified as follows:adenosine （38）,uridine（40）,kaemferol（42）,7,4’-dimethoxy-kaernkerol（43）,difructose（44）, sucrose（45）,glucose（46）,sitosterol（47）,daucosterol（48）.Among these compounds,compound 26,27,28 were new compounds.All the known compounds were isolated from Miliusa balansae for the first time except for 34. 3)Bioactivity research:Alkaloids and phenolic glycosides isolated from Miliusa balansae were assayed on anti-inflammatory activity by suppressing phosphodiesteraseâ… in vitro.Compounds 31,33,34,35,and 36 were enhanced phosphodiesteraseâ… activity,while compound 32 inhibited phosphodiesteraseâ… activity.Alkaloids and triterpenoid saponins isolated from Miliusa balansae were assayed on anti-arteriosclerosis activity by suppressing macrophage absorbing LDL. Compounds 5 and 41 showed some positive results on controlling macrophage’s absorption of LDL,suggesting the potential activity on anti-atherosclerosis.While compound 37 promoted the absorption of LDL by macrophage on the consistency of 0.2 mg/ml,which indicated that compound 37 promoted the formation of arteriosclerosis.Phenethanol glycosides and phenolic glycosides isolated from Ilex hainanensis and Miliusa balansae were assayed on the neuroprotective of PC12 stimulated by H₂O₂.The results revealed that compounds 27,31,and 33 showed some protection against H₂O₂-induced injury of PC12 cells,which implied that these compounds had the activity to protect the neruris system.Compound 29 showed cytotoxicity,the structures of 27 and 29 are different in the configuration of apiose,withα- andβ-configuration respectively,which indicated that theα- apiose may reinforce protection PC12 cells.4)The chemical constituents and bioactivity of genus Ilex and Miliusa were reviewed in this paper,which provide advice for further research of plants from these genuses.