疣海星生物活性成分研究
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摘要
以稻瘟霉模型进行活性追踪，应用大孔树脂、葡聚糖凝胶、正相硅胶、反相硅胶和高效液相等多种现代层析分离技术，对产于我国海南省三亚海域的疣海星(Pentaceraster sp．)的抗肿瘤与抗真菌活性成分进行了系统的分离纯化和生物活性研究，得到13个皂苷类化合物。应用现代波谱学技术和化学方法测定了其中7个化合物的化学结构，4个为新化合物，另3个为该物种中首次分离得到的化合物。
采用现代光谱和化学方法，特别是采用~1HNMR、~(13)CNMR和二维核磁共振技术(DQCOSY、TOCOSY、HMQC、HMBC和NOESY等)和电喷雾离子质谱，确定了7个化合物的化学结构，分别是：化合物Ps-1：3-O-[(2'-methoxyl)-xylopyranosides]-6β,8,16β,29-tetrahydroxy-5α-cholestanol-15α-O-sodium sulfonate (Pentacerasteroside A)，化合物Ps-2:6α-O-{β-D-quinovopyranosyl-(1→3)-β-D-galactopyranosyl-(1→4)-[[β-D-quinovopyranosyl-(1→2)]-β-D-fucopyranosyl-(1→3)-β-D-quinovopyranosyl}-16,22-epoxy-20-hydroxy-24-methyl-5α-cholest-9(11)-en-3β-O-sodium sulfonate(Pentacerasteroside B),化合物Ps-3: 24-O-[(2'-methoxyl)-β-D-xylopyranosyl-(1→2)-α-L-arabinofuranosyl]-3β,5,6α,8,15-pentahydroxy-5-cholestanol(Pentacerasteroside C),化合物Ps-4: 6α-O-{β-D-fucopyranosyl-(1→2)-β-D-quinovopyranosyl-(1→4)-[β-D-(3'-methoxyl)-quinovopyranosyl-(1→2)]-β-D-xy lopyranosyl-(1→3)-β-D-quinovopyranosyl}-20-methyl-25-methoxyl-5α-cholest-9(11),23-diene-22-one-3β-O-sodium sulfonate(Pentacerasteroside D),化合物Ps-5:6α-O-{β-D-fucopyranosyl-(1→2)-β-D-quinovopyranosyl-(1→4)-[β-D-(3'-methoxyl)-quinovopyranosyl-(1→2)]-β-D-xylopyranosyl-(1→3)-β-D-quinovopyranosyl}-20,25-dimethyl-5α-cholest-9(11),23-diene-22-one-3β-O-sodium sulfonate(Goniopectenoside A),化合物Ps-6: 6α-O-{β-D-fucopyranosyl-(1→2)-α-L-arabinopyranosyl-(1→4)-[β-D-quinovopyranosyl-(1→2)]-β-D-xylopyranosyl-(1→3)-β-D-quinovopyranosyl}-5α-pregn-9(11)-en-20-one-3β-O-sodium sulfonate(novaeguinoside C)和化合物Ps-7: 6α-O-{β-D-fucopyranosyl-(1→2)-β-D-galactopyranosyl-(1→4)-[β-D-quinovopyranosyl-(1→2)]-β-D-xylopyranosyl-(1→3)-β-D-quinovopyranosyl}-5α-pre gn-9(11)-en-20-one-3β-O-sodium sulfonate(asteronyl pentaglycoside sulfate)。其中化合物Ps-1，2，3，4为新化合物，分别命名为Pentacerasterside A，B，C和D。
体外细胞毒活性试验表明：2个海星皂苷类化合物Ps-4～5对P388小鼠白血病和BeL7402人肺癌等2种人肿瘤细胞株具有较强的细胞毒活性。抗真菌活性研究显示：化合物Ps-2，4～5有较强的抗真菌活性，化合物Ps-2对红色毛藓菌的MIC值为6.9ug／ml，对申克氏孢子丝菌MIC值为2.2ug／ml，对烟曲霉菌MIC值为4.1ug／ml，对白色念珠菌、新生隐球菌、热带念珠菌MIC值均为3.4ug／ml；化合物Ps-4～5对白色念珠菌、新型隐球菌、光滑念珠菌、近平滑念珠菌、热带念珠菌、红色毛藓菌和申克氏孢子丝菌MIC值在1.1～17.5ug／ml之间；化合物Ps-1对申克氏孢子丝菌的MIC值为8.8ug／ml。
本课题对疣海星的活性成分及药理作用进行了深入系统的研究，首次从该种中发现了多个具有生物活性的化学成分，为研制新的抗肿瘤和抗真菌药物提供了科学依据，对开发利用我国丰富的海洋生物资源具有重要的意义。研究成果丰富了海洋药物学和天然药物化学的研究内容，对今后进一步深入研究我国其它海洋生物的活性成分和药理作用具有借鉴意义。
Pentaceraster sp. distributed abundantly in South China Sea especially in Sanya Gulf, Hainan province. Up to date, only few chemical works have been performed on these starfishes. Guided by P. oryzae bioassay model, the anticancer and antifungal constituents of Pentaceraster sp. has been investigated by various chromatography methods including LPLC, MPLC, HPLC on resin, silica gel, ODS RP C-18 and Zobax SB C-18 respectively.
Thirteen compounds were isolated; Seven compounds were elucidated by chemical and spectral analysis(IR; EI-MS, HREI-MS, ESI-MS;~1HNMR, ~(13)CNMR, HMQC, TOCSY,~1H-~1H COSY, DQF-COSY, HMBC, NOESY).
Seven steroidal glycosides were isolated from the starfish Pentaceraster sp.(from Shanya Gulf, Hainan province) and elucidated as: 3-O-[(2'-methoxyl)-xylopyranosides]- 6β,8,16β,29-tetrahydroxy-5a-cholestanol-15α-O-sodium sulfonate(Ps-1), 6α-O-{β-D-quinovopyranosyl-(1→3)-β-D-galactopyranosyl-(1→4)-[β-D-quinovopyranosyl-(1→2)]-β-D-fucopyranosyl-(1→3)-β-D-quinovopyranosyl}-16,22-epoxy-20-hydroxy-24-methyl-5α-cholest-9(11)-en-3β-O-sodium sulfonate(Ps-2), 24-O-[(2'-methoxyl)-β-D-xylopyranosyl-(1→2) -α-L-arabinofuranosyl]-3β,5,6α,8,15-pentahydroxy-5-cholestanol(Ps-3), 6α- O-{β-D-fucopyranosyl-(1→2)-β-D-quinovopyranosyl-(1→4)-[β-D-(3'-methoxyl)-quinovopyranosyl-(1→2)]-β-D-xylopyranosyl-(1→3)-β-D-quinovopyranosyl}-20-methyl-25-methoxyl-5α- cholest -9(11),23-diene-22-one-3β- O-sodium sulfonate(Ps-4), 6α- O-{β-D-fucopyranosyl-(1→2)-β-D- quinovopyranosyl-(1→4)-[β-D-(3'-methoxyl)-quinovopyranosyl-(1→2)]-β-D-xylopyranosyl-(1→3)-β-D-quinovopyrano-syl}-20,25-dimethyl-5α- cholest -9(11),23-diene-22-one-3β- O-sodium sulfonate(Ps-5), 6a-O-{β-D-fucopyranosyl-(1→2)-α-L- arabinopyranosyl-(1→4)-[β-D-quinovopyranosyl-(1→2)]-β-D-xylopyranosyl-(1→3)-β-D-quinovopyranosyl}-5α-pregn-9(11)-en-20-one-3β-O-sodium sulfonate(Ps-6), 6α- O-{β-D-fucopyranosyl-(1→2)-β-D-galactopyranosyl-(1→4)-[β-D-quinovopyranosyl-(1→2)]-β-D-xylopyranosyl-(1→3)- β-D-quinovopyranosyl}-5α-pregn-9(11)-en-20-one-3β- O-sodium sulfonate(Ps-7).
The compounds showed moderate cytotoxity in vitro against many human tumor cell lines such as P388 and BEL7402. Ps-2 showed strongantifungalactivity against Candida albicans with MIC 3.4ug/ml, C. parapsilosis with MIC 1.7ug/ml, C. neoformans with MIC 3.4ug/ml, C. pseudotropicalis with MIC 3Aug/ml,Sporothrixschenckii with MIC 2.2ug/ml, Aspergillus fumigantus with MIC 4.1ug/ml, Trichophyton rubrum with MIC 6.9ug/ml.Ps-1,4～5 showed strong antifungal activity against Candida albicans,C. parapsilosis,C. neoformans, C. pseudotropicalis, Sporothrixschenckii, Aspergillus fumigantus, Trichophyton rubrum with MIC from 1.1ug/ml to 125ug/ml.
Our studies focused on bioactive constituents of Pentaceraster sp. established a foundation for further development of the kind starfish with abundant resources in South China Sea. We are in the process of applying for patents for the structures of Ps-1～7 and their remarkable cell growth inhibitory activity against human tumor cell lines.
采用现代光谱和化学方法，特别是采用~1HNMR、~(13)CNMR和二维核磁共振技术(DQCOSY、TOCOSY、HMQC、HMBC和NOESY等)和电喷雾离子质谱，确定了7个化合物的化学结构，分别是：化合物Ps-1：3-O-[(2'-methoxyl)-xylopyranosides]-6β,8,16β,29-tetrahydroxy-5α-cholestanol-15α-O-sodium sulfonate (Pentacerasteroside A)，化合物Ps-2:6α-O-{β-D-quinovopyranosyl-(1→3)-β-D-galactopyranosyl-(1→4)-[[β-D-quinovopyranosyl-(1→2)]-β-D-fucopyranosyl-(1→3)-β-D-quinovopyranosyl}-16,22-epoxy-20-hydroxy-24-methyl-5α-cholest-9(11)-en-3β-O-sodium sulfonate(Pentacerasteroside B),化合物Ps-3: 24-O-[(2'-methoxyl)-β-D-xylopyranosyl-(1→2)-α-L-arabinofuranosyl]-3β,5,6α,8,15-pentahydroxy-5-cholestanol(Pentacerasteroside C),化合物Ps-4: 6α-O-{β-D-fucopyranosyl-(1→2)-β-D-quinovopyranosyl-(1→4)-[β-D-(3'-methoxyl)-quinovopyranosyl-(1→2)]-β-D-xy lopyranosyl-(1→3)-β-D-quinovopyranosyl}-20-methyl-25-methoxyl-5α-cholest-9(11),23-diene-22-one-3β-O-sodium sulfonate(Pentacerasteroside D),化合物Ps-5:6α-O-{β-D-fucopyranosyl-(1→2)-β-D-quinovopyranosyl-(1→4)-[β-D-(3'-methoxyl)-quinovopyranosyl-(1→2)]-β-D-xylopyranosyl-(1→3)-β-D-quinovopyranosyl}-20,25-dimethyl-5α-cholest-9(11),23-diene-22-one-3β-O-sodium sulfonate(Goniopectenoside A),化合物Ps-6: 6α-O-{β-D-fucopyranosyl-(1→2)-α-L-arabinopyranosyl-(1→4)-[β-D-quinovopyranosyl-(1→2)]-β-D-xylopyranosyl-(1→3)-β-D-quinovopyranosyl}-5α-pregn-9(11)-en-20-one-3β-O-sodium sulfonate(novaeguinoside C)和化合物Ps-7: 6α-O-{β-D-fucopyranosyl-(1→2)-β-D-galactopyranosyl-(1→4)-[β-D-quinovopyranosyl-(1→2)]-β-D-xylopyranosyl-(1→3)-β-D-quinovopyranosyl}-5α-pre gn-9(11)-en-20-one-3β-O-sodium sulfonate(asteronyl pentaglycoside sulfate)。其中化合物Ps-1，2，3，4为新化合物，分别命名为Pentacerasterside A，B，C和D。
体外细胞毒活性试验表明：2个海星皂苷类化合物Ps-4～5对P388小鼠白血病和BeL7402人肺癌等2种人肿瘤细胞株具有较强的细胞毒活性。抗真菌活性研究显示：化合物Ps-2，4～5有较强的抗真菌活性，化合物Ps-2对红色毛藓菌的MIC值为6.9ug／ml，对申克氏孢子丝菌MIC值为2.2ug／ml，对烟曲霉菌MIC值为4.1ug／ml，对白色念珠菌、新生隐球菌、热带念珠菌MIC值均为3.4ug／ml；化合物Ps-4～5对白色念珠菌、新型隐球菌、光滑念珠菌、近平滑念珠菌、热带念珠菌、红色毛藓菌和申克氏孢子丝菌MIC值在1.1～17.5ug／ml之间；化合物Ps-1对申克氏孢子丝菌的MIC值为8.8ug／ml。
本课题对疣海星的活性成分及药理作用进行了深入系统的研究，首次从该种中发现了多个具有生物活性的化学成分，为研制新的抗肿瘤和抗真菌药物提供了科学依据，对开发利用我国丰富的海洋生物资源具有重要的意义。研究成果丰富了海洋药物学和天然药物化学的研究内容，对今后进一步深入研究我国其它海洋生物的活性成分和药理作用具有借鉴意义。
Pentaceraster sp. distributed abundantly in South China Sea especially in Sanya Gulf, Hainan province. Up to date, only few chemical works have been performed on these starfishes. Guided by P. oryzae bioassay model, the anticancer and antifungal constituents of Pentaceraster sp. has been investigated by various chromatography methods including LPLC, MPLC, HPLC on resin, silica gel, ODS RP C-18 and Zobax SB C-18 respectively.
Thirteen compounds were isolated; Seven compounds were elucidated by chemical and spectral analysis(IR; EI-MS, HREI-MS, ESI-MS;~1HNMR, ~(13)CNMR, HMQC, TOCSY,~1H-~1H COSY, DQF-COSY, HMBC, NOESY).
Seven steroidal glycosides were isolated from the starfish Pentaceraster sp.(from Shanya Gulf, Hainan province) and elucidated as: 3-O-[(2'-methoxyl)-xylopyranosides]- 6β,8,16β,29-tetrahydroxy-5a-cholestanol-15α-O-sodium sulfonate(Ps-1), 6α-O-{β-D-quinovopyranosyl-(1→3)-β-D-galactopyranosyl-(1→4)-[β-D-quinovopyranosyl-(1→2)]-β-D-fucopyranosyl-(1→3)-β-D-quinovopyranosyl}-16,22-epoxy-20-hydroxy-24-methyl-5α-cholest-9(11)-en-3β-O-sodium sulfonate(Ps-2), 24-O-[(2'-methoxyl)-β-D-xylopyranosyl-(1→2) -α-L-arabinofuranosyl]-3β,5,6α,8,15-pentahydroxy-5-cholestanol(Ps-3), 6α- O-{β-D-fucopyranosyl-(1→2)-β-D-quinovopyranosyl-(1→4)-[β-D-(3'-methoxyl)-quinovopyranosyl-(1→2)]-β-D-xylopyranosyl-(1→3)-β-D-quinovopyranosyl}-20-methyl-25-methoxyl-5α- cholest -9(11),23-diene-22-one-3β- O-sodium sulfonate(Ps-4), 6α- O-{β-D-fucopyranosyl-(1→2)-β-D- quinovopyranosyl-(1→4)-[β-D-(3'-methoxyl)-quinovopyranosyl-(1→2)]-β-D-xylopyranosyl-(1→3)-β-D-quinovopyrano-syl}-20,25-dimethyl-5α- cholest -9(11),23-diene-22-one-3β- O-sodium sulfonate(Ps-5), 6a-O-{β-D-fucopyranosyl-(1→2)-α-L- arabinopyranosyl-(1→4)-[β-D-quinovopyranosyl-(1→2)]-β-D-xylopyranosyl-(1→3)-β-D-quinovopyranosyl}-5α-pregn-9(11)-en-20-one-3β-O-sodium sulfonate(Ps-6), 6α- O-{β-D-fucopyranosyl-(1→2)-β-D-galactopyranosyl-(1→4)-[β-D-quinovopyranosyl-(1→2)]-β-D-xylopyranosyl-(1→3)- β-D-quinovopyranosyl}-5α-pregn-9(11)-en-20-one-3β- O-sodium sulfonate(Ps-7).
The compounds showed moderate cytotoxity in vitro against many human tumor cell lines such as P388 and BEL7402. Ps-2 showed strongantifungalactivity against Candida albicans with MIC 3.4ug/ml, C. parapsilosis with MIC 1.7ug/ml, C. neoformans with MIC 3.4ug/ml, C. pseudotropicalis with MIC 3Aug/ml,Sporothrixschenckii with MIC 2.2ug/ml, Aspergillus fumigantus with MIC 4.1ug/ml, Trichophyton rubrum with MIC 6.9ug/ml.Ps-1,4～5 showed strong antifungal activity against Candida albicans,C. parapsilosis,C. neoformans, C. pseudotropicalis, Sporothrixschenckii, Aspergillus fumigantus, Trichophyton rubrum with MIC from 1.1ug/ml to 125ug/ml.
Our studies focused on bioactive constituents of Pentaceraster sp. established a foundation for further development of the kind starfish with abundant resources in South China Sea. We are in the process of applying for patents for the structures of Ps-1～7 and their remarkable cell growth inhibitory activity against human tumor cell lines.
引文
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