Studies On The Bioactive Constituents From Holothuria Arenicola Semper

Holothuria arenicola Semper distributed abundantly in South China Sea. No chemical works have been performed on this sea cucumber up to date. Guided by Pyricularia oryzae bioassay model, the anti-cancer and anti-fungal constituents of Holothuria arenicola Semper have been investigated by various chromatography methods including LPLC, MPLC, HPLC on silica gel, ODS RP C18 and Zorbax SB C18 respectively. Eighteen compounds were isolated and structures of twelve 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 Holothuria arenicola Semper, twelve compounds were isolated and identified, including eight triterpene glycosides: 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-holosta-9(11)-ene-3β,12α,17α-triol (HA-1, holothurin A), 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]-holosta-22-hydroxyholost-9(11)-ene-3β,12α,17α-triol (HA-2, holothurin A1), 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-holosta-9(11)-ene-3β,12α-diol (HA-3, fuscoineroside C),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]-holosta-9(11)-ene-3β,12α,17α-triol (HA-4, echinoside A), 3-O-[3-O-methyl-β-D-glucopyranosyl-(1→3)-β-D-glucopyranosyl-(1→4) -β-D-quinovopyranosyl-(1→2)-4-O-sulfato-β-D-xylopyranosyl]-22,25-epoxy-holosta-9(11)-ene-3 β,12α,17α-triol ( HA-5, arenicolaside A ), 3-O-[ 3-O-methyl-β-D-glucopyranosyl-(1→3)-β-D- glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-4-O-sulfato-β-D-xylopyranosyl]-22,25-epoxy-holosta-9(11)-ene-3β,12α-diol (HA-6, arenicolaside B), 3-O-[β-D- quinovopyranosyl-(1→2)-4-O-sodiumsulfate-β-D-xylopyranosyl]-22,25-epoxy-holosta-9(11)-ene-3β,12α,17α-triol (HA -7, holothurin B), 3-O-[β-D-quinovopyranosyl-(1→2)-4-O-sodiumsulfate-β-D-xylopyranosyl]-- holosta-9(11)-ene- 3β,12α,17α-triol (HA-8, echinoside B); and four sterols: 3β- sulfato-24-methylenecholest- 5α- cholesta-7( 8 )-ene (HA-9, arenicolaside C), 3β-sulfato-24- methylenecholest-5α-cholesta ( HA-10, arenicolaside D), 3β-sulfato-24-methyl-5α- cholesta- 7,22E-diene (HA-11, arenicolaside E), 3β-sulfato-24-methyl-5α-cholesta-7(8)-ene (HA-12, arenicolaside F).Eight triterpene glycosides HA-1～HA-8 showed activity against P.oryzae. The preliminary cytotoxic assay of these saponins indicated that BEL-740, HL-60, A-549 and P388. In the anti-fungus screen, eight triterpene glycosides were confirmed active against several fungus lines.Our studies focused on bioactive constituents of Holothuria arenicola Semper 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.