Studies On The Bioactive Constituents From Holothuria Grisea Selenka

The sea cucumber Holothuria grisea Selenka abundantly distributed in the sea from South Florida through the Bahamas to Brazil.Two compounds holothurin A and holothurin A₁ have been isolated by Japanese scientists from this animal while the chemical constitutents and their pharmacological activitities of this sea cucumber still need to be further investegated.Guided by Pyricularia oryzae bioassay model,we isolated 12 compounds from Holothuria grisea Selenka using various chromatography methods including LPLC, MPLC,HPLC on silica gel,ODS RP C₁₈ and Zorbax SBC₁₈.The structure of 8 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.),their chemical structures were:HG-1:3β-O-[β-D-3-O-methylglucopyranosyl (1→3)-β-D-glucopyranosyl(1→4)-β-D-quinovopyranosyl(1→2)-β-D-xylopyranosyl (4→1)-β-D-glucopyranosyl]-22,25-epoxy-holost-9-(11)-ene-3β,12α-diol(nam ed griseasideA);HG-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]-22,25-epox y-holosta-9(11)-ene-3β,12α,17α-triol(holothurinA);HG-3:3-O-[3-O-methyl-β-D-glucopyranosyl -(1→3)-β-D-glucopyranosyl-(1→4)-β-D-quinovopyranosyl-(1→2)-4-O-sod iumsulfate-β-D-xylopyranosyl]-holosta-22-hydroxyholost-9(11)-ene-3β,12α,17α-triol(holo thurinA₁);HG-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]-holosta-9(1 1)-ene-3β,12α,17α-triol(echinosideA);HG-5:3β-O-[β-D-3-O-methylglucopy-ranosyl (1→3)-β-D-glucopyranssyl(1→4)-β-D-quinovopyranosyl(1→2)-β-D-xylopyranosyl(4→1)-β-D-glucopyranosyl]-holosta-22-hydroxyholost-9(11)-ene-3β,12α-diol(named griseasideB); HG-6:3-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,17α-triol(arenicolasideA);HG-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-xylpyra nosyl]-22,25-epoxy-holosta-9(11)-ene-3β,12α-diol(fuscoinerosideC);HG-8:3β-O-[β-D-3-O-methylglucopyranosyl (1→3)-β-D- glucopyrano syl(1β4)-β-D-quinovopyranosyl(1→2)-β-D-xylopyranosyl (4→1)-β-D-glucopyranosyl]-22,25-epoxy-holosta-9(11)-ene-3β,12α,17α-tr iol(holothurinoside A).All the triterpene glycosides HG-1～HG-8 showed activity against P.oryzae.The preliminary cytotoxic assay of these saponins indicated that HG-1～HG-8 exhibited significant cytot0xicity against BEL-740,HL-60,A-549 and Molt-4 cell lines.In the anti-fungus screen,eight tfiterpene glycosides were confirmed active against several fungus lines.Our studies focused on the bioactive constituents of Holothuria grisea Selenka have established a foundation for the further research and development of this kind of sea cucumber with abundant resources,and provided important leading compounds for the development of new anti-cancer and anti-fungus drugs.