Studies On The Bioactive Constituents From Sea Cucumber Holothuria Leucospilota And Holothuria Scabra

H. leucospilota and H. scabra Jaeger, are sea cucumber widely distributed in the South China Sea, along the coast of Hainan Province, China. Guided by P. oryzae bioassay model, the antitumor and antifungal constituents of these two kinds of sea cucumber have been investigated by various chromatography methods including LPLC, MPLC, HPLC on resin, silica gel, ODS RP C18 and Zobax SB C18 respectively, and twenty four compounds were isolated. Structures of twenty two compounds were elucidated by chemical and spectral analysis (IR, EI-MS, ESI-MS, HRESI-MS, 1H NMR, ¹³C NMR, ¹H-¹HCOSY, DQCOSY, TOCSY, HMQC, HMBC, NOESY, etc.).The structure of ten triterpene glycosides from the H. leucospilota were elucidated as:3-O-{β-D-quinovopyranosyl(1→2)-4'-O-sulfate-β-D-xylopyranosyl}-holosta-22,25- epoxy-9-ene-3β,12a,17a-triol(HL-1,holothurinB),3-O-{β-D-quinov-opyranosyl(1→2)-4'-O-sulfate-β-D-xylopyranosyl}-holosta-9-ene-3β,12a,17a,25a-tetraol(HL-2,HL-5),3-O- {β- D-quinovopyranosyl(1→2)-4'-O-sulfate-β-D-xylopyra-nosyl}-holosta-9-ene-23-keto- ne-3β,12a,17a,25a-tetraol(HL-3),3-O-{β-D-quinovo-pyranosyl(1→2) -4'-O-sulfate-β-D- xylopyranosyl}-holosta-9-ene-3β,12a,17a-triol (HL-4),3-O-{β-D-quinovopyranosyl(1→2)-4'-O-sulfate-β-D-xylopyranosyl}-holos-ta-9-ene-22-ketone-3β,12a,17a,25a-tetraol (HL-6,leucospilotaside A);3-O-{β-D-quinovopyranosyl(1→2)-4'-O-sulfate-β-D-xylopyr- anosyl}-holosta-9-ene-3β,12a,17a,22a-tetraol(HL-7), 3-O-{β-D-quinovopyranosyl(1→2) -4'-O-sulfate-β-D-xylopy-ranosyl}-holosta-9-ene-3β,12a,17a,24β-tetraol(HL-8, leucos- pilotaside B),3-O-{4'-O-sulfate-β-D-xylopyranosyl}-holosta-22,25-epoxy-9-ene-3β,12a, 17a-triol(HLB3b,leucospilotaside C),3-O-{3''''-O-methyl-β-D- glucopyranosyl(1→3)-β-D-glucopyranosyl(1→4)-β-D-quinovopyranosyl(1→2)-4'-O-sulfate-β-D-xylop yransyl} -holos-ta-22,25-epoxy-9-ene-3β,12a,17a-triol(HL-9,holothurinA). Desulfated structure of leucospilotaside A was elucidated as:O-{β-D-quinovopyranosyl(1→2)-β-D-xylop- yanosyl}-holosta-9-ene-22-ketone-3β,12a, 17a,25a-tetraol(DSHL-6,leucospilotaside a).The structure of ten triterpene glycosides from the H scabra Jaeger were elucidated as: 3-O-{3''''-O-methyl-β-D-glucopyranosyl(1→3)-β-D-glucopyranosyl(1→4)-β-D-qui- novopyranosyl(1→2)-4'-O-sulfate-β-D-xylopyransyl}-holosta-9-ene-23-ketone-3β,12a, 17a-triol(HS-1),3-O-{3''''-O-methyl-β-D-glucopyranosyl(1→3)-β-D-glucopyranosyl(1→4)-β-D-quinovopyranosyl(1→2)-4'-O-sulfate-β-D-xylopyransyl}-holosta-9,25-diene -3β,12a,17a-triol(HS-2,scabrasideA),3-O-{3''''-O-methyl-β-D-glucopyranosyl(1→3)-β-D-glucopyranosyl(1→4)-β-D-quinovopyranosyl(1→2)-4'-O-sulfate-β-D-xylopyran- syl}-holosta-9,24-diene-3β,12a,17a-triol(HS-3),3-O-{3''''-O-meth-yl-β-D-glucopyrano-syl(1→3)-β-D-glucopyranosyl(1→4)-β-D-quinovopyranosyl(1→2)-4'-O-sulfate-β-D- xylopyransyl}-holosta-9-ene-3β,12a,17a-triol(HS-4),3-O-{3''''-O-methyl-β-D-glucopy- ranosyl(1→3)-β-D-glucopyranosyl(1→4)-β-D-quinovopyranosyl(1→2)-4'-O-sulfate-β-D-xylopyransyl}-holosta-9-diene-3β,12a,17a,22a-tetraol(HS-5,holothurinA1), holothurinA(HS-6),3-O-{3''''-O-methyl-β-D-glucopyranosyl(1→3)-β-D-glucopyranosyl(1→4)-β-D-quinovopyranosyl(1→2)-4'-O-sulfate-β-D-xylopyransyl}-holosta-9-ene-22- ketone-3β,12a,17a-triol(HS-7,scabrasideB),3-O-{3''''-O-methyl-β-D-glucopyranosyl (1→3)-β-D-glucopyranosyl(1→4)-β-D-quinovopyranosyl(1→2)-4'-O-sulfate-β-D-xylo-pyransyl}-holosta-9,23-diene-3β,12a,17a-triol(HS-8),3-O-{3''''-O-methyl-β-D-glucopy- ranosyl(1→3)-β-D-glucopyranosyl(1→4)-β-D-quinovopyranosyl(1→2)-4'-O-sulfate-β- D-xylopyransyl}-holosta-9-ene-3β,12a,17a,25a-tetraol(HS-10),3-O-{3''''-O-methyl-β- D-glucopyranosyl(1→3)-β-D-glucopyranosyl(1→4)-β-D-quinovopyranosyl(1→2)-4'-O-sulfate-β-D-xylopyransyl}-holosta-22,25-epoxy-9-ene-3β,12a-diol(HS-11).All triterpene glycosides showed activity against P.oryzae.HS-3, HS-4and HS-5 exhibited significant cytotoxicity against HL-60, MOLT-4 leukemia cell lines respectively; while HL-1and HL-9 exhibited significant cytotoxicity against BEL-7402 cell line.HS-1^HS-5, HS-7, HS-8 and HL-4 showed significant active against seven fungi lines,and HS-3 against seven fungi lines have with MIC80 1 2, 4μg/ml.Our studies focused on bioactive constituents of H. leucospilota and H. scabra Jaeger , and we have established a foundation for further development of the sea cucumber with abundant resources in South China Sea,and provided important leading compounds for the development of new antitumor and antifungus drugs.