Studies On The Bioactive Constituents From Holothuria Axiloga, Thyone Anomala And Actinpyga Miliaris

Holothuria axiloga, Thyone anomala and Actinopyga miliaris distributed abundantly in South China Sea. Guided by Pyricularia oryzae bioassay model, the anti-cancer and anti-fungal constituents of Holothuria axiloga, Thyone anomala and Actinopyga miliaris have been investigated by various chromatography methods including LPLC, MPLC, HPLC on silica gel, ODS RP C18 and Zorbax SB C18 respectively. Forty-two compounds were isolated and structures of all the compounds were elucidated by chemical and spectral analysis (IR, EI-MS, ESI-MS, HRESI-MS, 1HNMR, 13CNMR, 1H-1H COSY, DQCOSY, TOCSY, HMQC, HMBC, NOESY, etc.).From the ethanolic extracts of H. axiloga, twenty triterpene glycosides were isolated and the structures were identified as the below, HA-1:3-O-{(3-O-methyl-β-D-glucopyra nosyl)-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-xylopyranosyl-(1→2)-[(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)]-β-D-xylopyranosyl}-23-acetoxy-7(8),25(26)-holostene-3β-ol(stichlorosideA2);HA-2:3-O-{(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-xylopyranosyl-(1→2)-[(3-O-methyl-β-D-glucop-yranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)]-β-D-xylopyranosyl}-23-acetoxy-7(8)-hol-ostene-3β-ol(stichlorosideA1);HA-3:3-O-{(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-xylopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-[(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)]-β-D-xylopyranosyl}-23-acetoxy-7(8),25(26)-holo-stene-3β-ol(stichlorosideC2);HA-4:3-O-{(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-xylopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-[(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)]-β-D-xylopyranosyl}-23-acetoxy-7(8)-holostene-3β-ol(stichlorosideC1);HA-5:3-O-[β-D-quinovopyranosyl-(1→2)-4-O-sulfo-β-D-xylopyranosyl]-22,25-epoxy-9(11)-holostene-3β,12α,17α-triol(axilogosideA);HA-6:3-O-[β-D-quinovopyranosyl-(1→2)-4-O-sodiumsulfato-β-D-xylopyranosyl]-22,25-epoxy-9(11)-holostene-3β,12α,17α-triol(holothurinB);HA-7:3-O-{(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyran-osyl-(1→4)-β-D-quinovopyranosyl-(1→2)-[(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)]-β-D-xylopyranosyl}-22,25-epoxy-9(11)-holostene-3β,12α,17α-triol(marmorosideG);HA-8:3-O-[(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-4-O-sodiumsulfato-β-D-xylopyranosyl]-22,25-epoxy-9(11)-holostene-3β,12α,17α-triol(holothurinA);HA-9:3-O-[(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-4-O-sodiumsulfato-β-D-xylopyranosyl]-9(11)-holostene-3β,12α-diol(pervicosideC);HA-10:3-O-[(3-O-methyl- β-D-glucopyranosyl)-(1→3)-β-D-glu-copyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-4-O-sodiumsulfato-β-D-xylopyranosyl]-22-one-9(11)-holostene-3β,12α-diol(fuscocinerosideB);HA-11:3-O-[(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-4-O-sodiumsulfato-β-D-xylopyranosyl]-22-one-25-acetoxy-9(11)-holostene-3β,12α-diol(fuscocinerosideA);HA-12:3-O-[(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-4-O-sodiumsulfato-β-D-xylopyranosyl]-22,25-epoxy-9(11)-holostene-3β,12α-diol(fuscocinerosideC);HA-13:3-O-{(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→4)]-β-D-xylopyranosyl}-23-acetoxy-9(11),24(25)-holostene-3β,12α-diol(axilogosideC);HA-14:3-O-[(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-4-O-sodiumsulfato-β-D-xylop-yranosyl]-9(11)-holostene-3β,12α,17α,22-tetrol(holothurinA1);HA-15:3-O-[(3-O-me-thyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-4-O-sodiumsulfato-β-D-xylopyranosyl]-9(11)-holostene-3β,12α,7α-triol(echinosideA);HA-16:3-O-[(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-4-O-sodiumsulfato-β-D-xylopyranosyl]-25-acetoxy-9(11)-holostene-3β,12α-diol(pervicosideA);HA-17:3-O-{(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-glucopyranosyl-(1→2)-[(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)]-β-D-xylopyranosyl}-23-acetoxy-7(8)-holostene-3β-ol(axilogosideG);HA-18:3-O-{(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-xylopyranosyl-(1→4)-β-D-xylopyranosyl-(1→2)-[(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)]-β-D-xylopyranosyl}-23-acetoxy-7(8)-holostene-3β-ol(axilogosideB);HA-19:3-O-{(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→4)]-β-D-xylopyranosyl}-23-acetoxy-9(11),24(25)-holostene-3β,12α-diol(axilogosideC);HA-20:3-O-{(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-[(3-O-methyl-β-D-glucopyranosyl)-(1→4)]-β-D-xylopyranosyl}-23-acetoxy-9(11)-holostene-3β,12α-diol(axilogosideH).From the ethanolic extracts of Thyone anomala, six compounds were isolated and identified, and the structures were identified as the below, TA-1: 3-O-[3-O-methyl -β-D-glucopyranosyl-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-4-O-sodiumsulfate-β-D-xylopyranosyl]-22,25-epoxy-holostene-9(11)-ene-3β,12α,17α-triol(holothurinA);TA-2:3-O-[3-O-methyl-β-D-glucopyranosyl-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-4-O-sodiumsulfate-β-D-xylopyranosyl]-22-hydroxyholos t-9(11)-ene-holostene-3β,12α,17α-triol(holothurinA1);TA-3:3-O-[3-O-methyl-β-D-glucopyranosyl-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-4-O-sodiumsulfate-β-D-xylopyranosyl]-22,25-epoxy-9(11)-ene-holostene-3β,12α-diol(anomalosideA);TA-4:3-O-[3-O-methyl-β-D-glucopyranosyl-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-4-O-sodiumsulfate-β-D-xylopyranosyl]-9(11)-ene-holostene-3β,12α,17α-triol(echinosideA);TA-5:3-O-[β-D-quinovopyranosyl-(1→2)-4-O-sodiumsulfate-β-D-xylopyranosyl]-22,25-epoxy-9(11)-ene-holostene-3β,12α,17α-triol(holothurinB);TA-6:3-O-[β-D-quinovopyranosyl-(1→2)-4-O-sodiumsulfate-β-D-xylopyranosyl]-9(11)-ene-holostene-3β,12α,17α-triol(echinoside B).From the ethanolic extracts of Actinopyga miliaris, sixteen compounds were isolated and identified, and the structures were identified as the below, AM-1:3-O-{(3-O-methyl-β- D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-[(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)]-β-D-xylopyranosyl}-9 (11),24(25)-holostene-3β,12α,22-triol(miliarisideA);AM-2:3-O-{(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→4)]-β-D-xylopyranosyl}-23-acetoxy-9(11),24(25)-holostene-3β,12α,17α-triol(miliarisideB);AM-3:3-O-{(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyra-nosyl-(1→2)-[(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)]-β-D-xylopyranosyl}-24-oxo-9(11)-holostene-3β,12α-diol(miliarisideC);AM-4:3-O-{(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-[(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)]-β-D-xylopyranosyl}-16-acetoxy-9(11),24(25)-holostene-3β,12α,17α-triol(miliarisideD);AM-5:3-O-{(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyra-nosyl-(1→2)-[(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)]-β-D-xylopyranosyl}-9(11)-holostene-3β,12α-diol(bivittosideD);AM-6:3-O-[3-O-methyl-β-D-glucopyranosyl-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-4-O-sodiumsulfate-β-D-xylopyranosyl]-22,25-epoxy-9(11)-ene-holostene-3β,12α,17α-triol(holothurinA);AM-7:3-O-[3-O-methyl-β-D-glucopyranosyl-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-4-O-sodiumsulfate-β-D-xylopyranosyl]-9(11)-ene-holostene-3β,12α,17α-triol(echinosideA);AM-8:3-O-[β-D-quinovopyranosyl-(1→2)-4-O-sodiumsulfate-β-D-xylopyranosyl]-22,25-epoxy-9(11)-ene-holostene-3β,12α,17α-triol(holothurinB);AM-9:3-O-[β-D-quinovopyranosyl-(1→2)-4-O-sodiumsulfate-β-D-xylopyranosyl]-9(11)-ene-3β,12α,17α-triol(echinosideB);AM-10:3-O-{(3-O-methyl-β-D-glucopyra nosyl)-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-[(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)]-β-D-xylopyranosyl}-9(11),24(25)-holostene-3β,12α-diol(miliarisideE);AM-11:3-O-{(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→4)]-β-D-xylopyranosyl}-23-acetoxy-9(11),24(25)-holostene-3β,12α-triol(miliarisideE);AM-12:3-O-{(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-[(3-O-methyl-β-D-glucopyranosyl)-(1→3)-β-D-glucopyranosyl-(1→4)]-β-D-xylopyranosyl}-25-acetoxy-9(11)-holostene-3β,12α-diol(miliariside F). And four sterols:AM-13:3β-sodiumsulfate-5α-cholesta-7(8),24-diene;AM-14:3β-sodiumsulfate-24-methyl-7(8)-ene-5α-cholesta;AM-15:3β-sodiumsulfate-7(8)-ene-5α-cholesta; AM-16: 3β- sodiumsulfate-5α-cholesta.The preliminary cytotoxic assay of these saponins indicated that BEL-740, HL-60, A-549 and P388. In the anti-fungus screen, six triterpene glycosides were confirmed active against several fungus lines.Our studies focused on bioactive constituents of Holothuria axiloga, Thyone anomala and Actinopyga miliaris have established a foundation for further research and development of the kind sea cucumber with abundant resources in South China Sea, and provided important leading compounds for the development of new anti-cancer and anti-fungus drugs.