Studies On Chemical Constituents And Pharmacological Effects Of Brucea Javanica(L.) Merr. Seeds

Brucea javanica(L.) Merr.(Simaroubaceae), also called laoyadan,kuzhenzi, kushenzi in China, is widely distributed in tropical and subtropicalof Asia, Indonesia and Australia. It’s mainly grown in Guangdong andGuangxi Provinces in China. The chemical compounds isolated from the seedsincluding quassinoids, flavonoids, triterpenoids and alkaloids. Thesechemicals show a wide spectrum of biological effects, such as antitumor,antimalarial, anti-inflammatory and antiviral etc. The antitumor effect ofquassinoids is the main research object for many scholars. We studied thechemical compositions of CH2Cl2 and Et OAc soluble extracts of Bruceajavanica and tested their biological effects.Objective: The chemical constituents of the seeds of Brucea javanicawere studied by using column chromatography, preparative thin layerchromatography and Sephadex LH-20 column chromatography, and allcompounds isolated from Brucea javanica were identified by UV, 1D and 2DNMR, MS and so on. In further research, we studied their antitumor activity invitro by MTT methods.Methods: The dry seeds of Brucea javanica(12 Kg) were extracted with95% Et OH under heated reflux, and then the extract was dispersed in H2 O andpartitioned with petroleum ether(Pet), dichloromethane(CH2Cl2) and ethylacetate(Et OAc) successively. The parts of CH2Cl2 and Et OAc were isolatedand purified by silica gel column chromatography, preparative thin layerchromatography, Sephadex LH-20 column chromatography and reverse phasesilica gel column chromatography. Structures of pure compounds wereelucidated with UV, 1H-NMR, 13C-NMR, 1H-1H COSY, HMBC, HMQC,NOESY, ROESY and MS. The inhibition effects to Bel-7402, SMMC-7721 and Hep G2 cell lines were further studied by MTT methods in last.Adriamycin, cisplatin, taxol and methotrexate were used as positive control.Results: Twenty-seven compounds were isolated from the seeds of Bruceajavanica, and twenty-three of them were identified, including nine quassinoids:bruceine A(3), bruceine B(4),(+)-dehydrobruceantinol(10), bruceantinol(11), bruceaketolic acid(12), bruceine C(15), bruceine D(16), yadanziosideG(17), dehydrobruceine B(22). Five triterpenoids: β-stigmasteryl 3-O-β-D-glucopyranoside(1), β-sitosteryl β-D-glucoside(2), β- sitosterol(5),stigmasta- 4, 22- diene- 3β, 6β-diol(8), 7-hydroxystigmasterol(9).Three flavonoids: luteolin(18), chrysoeriol(19), apigenin(20). Three benzoicacid esters: vanillic acid(14), syringic acid(21), gallincin(24). Two nitrogencompounds: 1H-indole-3-carbonxal-dehyde(7), 2, 4-dihydroxy pyrimidine(26). A nitrogen compound: dehydrochebulic acid trimethyl ester(23).Compounds 3, 4 and 15 showed strong inhibit activity to three tumor celllines(Bel-7402, SMMC-7721 and Hep G2 cell lines). When treated withcompound 3(concentration of 100 μmol/L), the survival rate of Bel-7402 celllines was about 25%, SMMC-7721 cecll lines was lower than 5%, and Hep G2 cell lines was about 50%. When treated with compound 4(concentration of100 μmol/L), the survival rate of three lines was about 15%, 5% and 50%,respectively. When treated with compound 15(concentration of 100 μmol/L),the survival rate of three lines was about 20%, 5% and 50%, respectively.Conclusion: Twenty-seven compounds were isolated and twenty-three ofthem were identified. Compounds 1, 2, 7, 8, 9, 20, 24, 26 were isolated fromBrucea javanica for the first time. Through the antitumor activities test, weknow that all of the three compounds were active to three tumor cell lines. Thesurvival rate of SMMC-7721 cells treated respectively with three isolatedcompounds were lower than 5%, and is far more lower than treated withadriamycin, cisplatin, taxol and methotrexate.