| This paper is composed of two parts:Antitumor activity of the compositions from Chlorophytum Laxum and Antitumor activity of alkaloids from Melicope ptelefolia.Objective:The main purpose of Chinese medicine chemistry is to find and discover the chemical constituents of Chinese medicine with pharmacological activity and to clarify the basis of medicinal plants.There is a rich resource of traditional Chinese medicine in our country,drugs classics from forefathers play a guiding role for the research and development of traditional Chinese medicine.The subject choosed two kinds of medicinal plants,which were not enough perfect for modem research:Liliacea,Chlorophytum,the root of Chlorophytum Laxum R.Br;Rutaceae,Melicope,stems and branchs of Melicope ptelefolia(Champ.Ex Benth)Hartley,researching antitumor activity of the compositions from Chlorophytum Laxum and antitumor activity of alkaloids from Melicope ptelefolia to provide the basis for clinical application of two kinds of medicinal plants.Chlorophytum Laxum R.Br belongs to Chlorophytum of Liliacea,also known as Sanjiaocao,Shuhuadiaolan(flora of China),shanjiucai(A Checklist of Vascular Plants of Guangxi).There are about 100 species plants from this genus,distributed mainly in tropical zone of Africa and Asia.There are four species plants from this genus in our country,which are distributed mainly in the southwest and Guangdong and Guangxi region.Chlorophytum Laxum R.Br is the poisonous plant,which shows pharmacological activities such as febrifugal,acesodyne,detumescence.In the previous studies,Our research group found ethanol extract of Chlorophytum Laxum showed strong inhibition activity to nasopharyngeal carcinoma cell lines and gastric cancer cell lines.But there was not more indepth research for the the basic,We don't know which some components play a role of antitumor.The mechanism of components showed strong antitumor activity,is still unknown,which will not be conducive to further to the development and utilization of Chlorophytum Laxum.In addition,there is few literature about active ingredient research of Chlorophytum Laxum both at home and abroad.Our research group used to report the research about chemical composition and extract of Chlorophytum Laxum.On the basis of preliminary experiment,we will further study the chemical compositions of Chlorophytum Laxum root and their antitumor activity,to clear antitumor active ingredient,which can provide the basis for the comprehensive utilization of Chlorophytum Laxum.Melicope ptelefolia(Champ.Ex Benth)Hartley belongs to Melicope of Rutaceae,in"lingnan herbalism record",also known as Sanyaku,Sanchaku,Sanzhiqiang,and so on,widely distributed in northeast to southwest in China,It is a commonly used Chinese herbal medicine,which showed pharmacological activities such as febrifugal,dehumidification.It was used in clinical to cure sore throats,malaria,icteric model hepatitis,rheumatism bone pain,and so on.The medicinal material is developed as Chinesepatent medicines,the clinical application is more extensive.There are a few leratures about alkaloids components and their active research,which reported the activity of the alkaloids in many aspects,such as inhibition of cholinesterase,which is beneficial to improve Parkinson's syndrome;sore throat,and used for the prevention and treatment of influenza,epidemic cerebrospinal meningitis and hepatitis B.It showed that Melicope ptelefolia as medicinal plant has important medicinal value.Alkaloids from Melicope ptelefolia are mainly quinoline alkaloids,a few of literatures reported the kind of alkaloids had extensive antitumor activity,but the literatures about antitumor activity study of alkaloids from Melicope ptelefolia reported less.In early experiments,we found four alkaloids were isolated from Melicopepetelfolia showed strong antitumor activity to some tumor cell lines.This paper focus on researching the compositions and antitumor activity of alkaloid from Melicope ptelefolia which is beneficial to clear antitumor activity of alkaloid from Melicope ptelefolia.There will be the extremely vital significance for the clinical application or secondary development of the medicinal material.Method:Isolate chemical compositions from two kinds of plant and research their antitumor activity.1.Isolate chemical compositions from Chlorophytum Laxum and research their antitumor activity.Dry root of Chlorophytum Laxum(1.2 Kg)were extracted with 95%ethanol by heating reflux extraction three times,after removal of solvent under reduce pressure to obtain the 95%EtOH crude extract.The crude extract was suspened by water,then petroleum ether,ethyl acetate,n-butanol was used to multiple extract compositions from the water in 2 times volume in turn.After removal of solvent under reduce pressure to obtain the petroleum ether crude extract(11 g),ethyl acetate crude extract(27g),n-butanol crude extract(30 g).ethyl acetate crude extract(27g)and n-butanol crude extract(30 g)were subjected to chromatography on silica gel,sephadexLH-20,semipreparative HPLC and recrystallization to monomer composition.Integrated use spectroscopic methods(UV,IRm MS,NMR)combination of the physical and chemical properties to confirm the structure of chemical composition.Literature reported that ethanol extract of Chlorophytum Laxum showed inhibition activity of human nasopharyngeal carcinoma cell line and human gastric carcinoma cell line,therefore,on the basis of study on chemical composition,detecting antitumor activity of the compositions.CCK8 method is used to screening antitumor activity of the compositions.2.Isolate alkaloids from Melicope ptelefolia and research their antitumor activity.Dried stems and barks of Evodia Lapta(30Kg)were extracted with 95%EtOH at room temperature six times.After removal of solvent under reduce pressure to obtain the 95%EtOH crude extract(885g).Using acid extraction and alkali precipitation to extract alkaloids of the EtOH crude extract,then used the column chromatography to isolate alkaloids.Firstly,the EtOH crude extract(875g)was suspended in acid water(HC1 pH=3)and then filtrated.The filtrate was disposed with alkaline liquor(NH3[aq]pH=10),then extract 10 times by the chloroform.After removal of solvent under reduce pressure to obtain the crude alkaloid 1(30,0g);Aqueous alkali was adjusted pH to 13 with NaOH,hen extract 10 times by the chloroform.After removal of solvent under reduce pressure to obtain the crude alkaloid 2(2.5g).According to the results of TLC,combine the crude alkaloids 1 and the crude alkaloids 2.The crude alkaloids(32.5g),reserved sample(2.5g),the crude alkaloids(30 g)were subjected to chromatography on silica gel,sephadexLH-20,semipreparative HPLC and recrystallization to monomer composition.Integrated use spectroscopic methods(UV,IR,MS,NMR)combination of the physical and chemical properties to confirm the structure of chemical composition.Researche suggested that alkaloids of Melicope ptelefolia showed antitumor activity,therefore,on the basis of study on chemical composition,detecting antitumor activity of the alkaloids.MTT method is used to screening antitumor activity of the compositions.Results:A total of 27 compounds were isolated from ethanol extract(185 g)of Chlorophytum Laxum,the structural of 25 compounds were identified,including a new compound:25-R-spirosta-3,5-dien-12?-ol(Compound 1),hendecane(Compound 2),methyl palmitate(Compound 3),butyl-isobutyl-phthalate(Compound 4),tridecanol(Compound 5),?-sitosterols(Compound 6),heptadecan-1-ol(Compound 7),1-octacosanol(Compound 8),stigmasterol(Compound 9),n-dotriacontanol(Compound 10),octadecane(Compound 11),nepethalate A(Compound 12),bis(2-ethyhe xyl)benzene-1,2-dicarboxylate(Compound 13),hentriacontane(Compound 14),Estigmasterol-3-O-?-D-glicopiranosideo(Compound 15),Palmitic monoglycerol ester(Compound 16),Heneicosane(Compound 17),Pentatriacontane(Compound 18),pentacosane(Compound 19),bis(2-ethyloctyl)phthalate(Compound 20),Apigenin-6-C-glucosylglucoside(Compound 21),7-2"-di-O-?-glucopyranosylisovitexin(Compound 22),o-phthalic acid bis-(2-ethyl decyl)-ester(Compound 23),3-O-?-authemisol(Compound 24),Diosgenin(Compound 25).CCK8 method is used to screening antitumor activity of ten compositions(Compounds 1?4?6?9?12?15?20?21?24?25).Theresults showed that 5-D-spirosta-3,5-dien-12??ol(compound 1),Apigenin-6-C-glucosylglucoside(compound 21),Diosgenin(compound 25)to activity of human nasopharyngeal carcinoma cell line(5-8F)had strong inhibitory effect,inhibitory effect of other compounds were not obvious.Apigenin-6-C-glucosylglucoside(compound 21)had significant inhibitory activity to gastric cancer cell line(BGC),5-D-spirosta-3,5-dien-12?-ol(compound 1)and Diosgenin(compound 25)showed strong inhibitory effect,inhibitory effect of other compounds were not obvious.A total of 11 alkaloids were isolated from Melicope ptelefolia:(-)-R-geibalansine(co-mpound 1),evolitrine(compound 2),skimmianine(compound 3),Kokusaginine(compound-4),7-hydroxydictamnine(compound 5),4-O-methyl-qumnolin-2-one(compound 6),4-meth-oxy-2(1H)-quinolinone(compound 7),R-(+)-platydesmin(componnd 8),dictamnine(com-pound 9),atamin(compound 10),N-P-trans-Coumaroyltyramine(compound 11).This paper showed the inhibition activity of alkaloids to Human breast cancer cells MCF-,Human colon cancer cells SW-480,Human cervical cancer HeLa cells.7-hyd roxydict amnine(IC50 8? pmol/L)and(-)-R-geibalansine(IC50 8?mol/L)showed very strong inhibitory activity to Human breast cancer cells MCF-7,R-(+)-Platydesmin(IC5012?mol/L)4-methoxy-2(1H)-quinolinone(IC50 14?mol/L)showed moderate intensity inhibitory activity,inhibitory activity of other alka loids was weak;(-)-R-geibalansine(IC508?mol/L)showed very strong inhibitory activity to Human colon cancer cells SW-480,dictamnine(IC50 10?mol/L),7-hydroxydictamnine(IC50 12 ?mol/L),R-(+)-platydesm(IC50 14 ?mol/L)showed moderate intensity inhibitory activity,inhibitory activity of othe ralka loids was weak;R-(+)-Platydesmin(IC50 8 ?mol/L),(-)-R-geibalansine(IC50 8?mol/L)showed very strong inhibitory activi ty to Human cervical cancer HeLa cells,dictamnine(IC50 12?mol/L),7-hydroxydictamnine(IC50 10?mol/L),showed moderate intensity inhibitory activity,inhibitory activity of othe ralka loids was weak.Conclusion:A total of 27 compounds were isolated from ethanol extract(185g)of Chlorophytum Laxum,the structural of 25 compounds were identified,including six steroids and glycoside,6 alkane compounds,5 benzoate compounds,4 senior fatty alcohols,2 fatty acids,2 flavor noid glycosides compounds,compound 1 was an new compound.15 compounds(compou-nds 3,4,5,7,8,10,12,13,15,16,20,21,22,23,24)were isolated from Chlorophytum Laxum for the first time.9 compounds(compounds 3,4,12,13,14,16,20,21,22)were isolated from this genus for the first time.The result showed antitumor activity of monomer chemical composition from Chlorophytu m Laxum,we can further define for the structures of the diosgenin and flavonoid glycosides,which have strong antitumor activity.A total of 11 alkaloids were isolated from Melicope ptelefolia,The 11 alkaloids belong to different alkaloids structure type,including 5 furoquinoline,4 quinolineketone,1 pyranoq uinoline,1 pyrrolidine.7 alkaloids were isolated from Melicope ptelefolia for the first time:(-)-R-geibalansine(compound 1),7-hydroxydictamnine(compound 5),4-0-methyl-quinolin-2-one(compound 6),4-methoxy-2(1 H)-quinolinone(compound 7),R-(+)-platydesmin(compound 8),atamin(compound 10),N-p-trans-Coumaroyltyramine(compound 11).The result showed a few of alkaloids from Melicope ptelefolia were quinoline.Study antitumor activity of alkaloids from Melicope ptelefolia in vitro,the alkaloids showed antitumor activity,but different alkaloid,antitumor activity is different,talkaloids with hydroxyl in the structure of quinoline showed stronger antitumor activity. |
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