狭叶五味子中化合物phyllocoumarin和（-）-epicatechin体外抗HBV活性的初步研究
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摘要
【目的】
联合用药比单一用药更能降低HBV感染者的发病率和死亡率,但长期联合用药导致耐药、副作用限制了抗HBV药物的使用。因此,不断地开发新的抗HBV药物迫在眉睫。目前临床抗HBV药物均为核苷类药物,如拉米夫定、阿德福韦酯、更昔洛韦、法昔洛韦、恩替卡韦和恩曲他滨,等。从天然资源中寻找新的抗HBV药物或先导化合物的研究是国内外新药研制中非常活跃的领域之一,也是我国近期创制新药的一条捷径。在过去20多年里,从天然资源中分离和鉴定出许多抗HBV药物。我国科学家发现北五味子(Schisandra chinensis)粗提物及其多种有效成分作用于多靶点,具有保肝解毒、抗氧化和抗HBV病毒等多方面的作用,并研发成功联苯双脂和双环醇两种具有我国自主知识产权的治疗肝炎药物。通过对狭叶五味子(Schisandra lancifolia)茎叶的化学成分和抗病毒活性研究,我们发现了Lancifodilactone F和Lancifodilactone G等多个具有抗HIV-1活性的化合物。本文对来源于狭叶五味子的化合物phyllocoumarin和(-)-epicatechin对抗HBV活性进行初步研究。
【方法】
筛选研究从狭叶五味子(Schisandra lancifolia)中分离化合物phyllocoumarin (1)和(-)-epicatechin(2)的体外抗HBV活性,以HepG2.2.15细胞为体外研究HBV模型,分别用RPMI-1640完全培养基稀释不同浓度的化合物1和2,与HepG2.2.15细胞共培养3天后收集上清,采用MTT法检测药物对HepG2.2.15细胞的生长影响和ELISA法检测培养上清中HBV HBsAg和HBeAg的含量,评价化合物1和2对HBV复制的影响。
【结果】
化合物1和2具有一定的体外抗HBV活性,其细胞毒性低,CC50均大于200μg/ml。化合物1具有较强的抑制HBsAg和HBeAg分泌作用。阳性对照药物阿德福韦酯也抑制HBV HBsAg和HBeAg分泌,但在相同浓度(1.6μg/ml)下其抑制作用较化合物1弱。
【结论】
phyllocoumarin和(-)-epicatechin能抗HBV活性,且毒性小。
【Objective】
Although the combination of the chemical drugs in the antiviral therapy more effectively reduce morbidity and mortality in HBV infected individuals than the use of single drug, emergence of HBV drugs resistance and side effects for long-term antiretroviral treatment limited the use of these drugs. So continuously development of new anti-HBV drugs appears to be inevitable. Presently, drugs of the anti-HBV therapy used clinically are nucleoside analogues.such as lamivudine, adefovir dipivoxil,ganciclovir, famciclovir and entecavir,emtricitabine. Natural products as the most consistently successful source in drug discovery may offer opportunities for finding anti-HBV drugs or lead compounds. In past 20 years, many natural compounds with anti-HBV activity have been isolated and indentified, In this study, we tested the anti-HBV effects of compounds isolated from Schisandra lancifolia. Scientists in China have found that compouds and many effective ingredients from Schisandra chinensis, with the function of protecting liver and decreasing levels of ALT, antioxidant and anti-HBV activity, tagarted many sites of virus. They have succeeded in developing two anti-HBV drugs, Bifendate and bicyclol. To confirm chemical compositions from Schisandra lancifolia and to evaluate its antiviral therapy,we have found many compouds, such as Lancifodilactone F and Lancifodilactone G, were against HIV-1. Here, we primarily studied that compouds phyllocoumarin (1) and (-)-epicatechin (2) from Schisandra lancifolia were anti-HBV activity.
【Methods】
To evaluate anti-HBV activities of phyllocoumarin (1) and (-)-epicatechin (2) from Schisandra lancifolia in vitro,we used HepG2.2.15 cells as HBV model in vitro. Different concentrations of compounds (1) and compounds (2) were added to the wells with the growth of HepG2.2.15 cells and the supernatant was collected at the third day. The cytotoxic activity on the growth of HepG2.2.15 cells was detected by MTT. ELISA for HBsAg and HBeAg assays were used to evaluate effects of compounds 1 and 2 on HBV replication.
【Result】
Both coupound 1 and compound 2 showed anti-HBV activity and exerted cytotoxicity against HepG2.2.15 cells with CC50>200μg/ml. But coupound 1 more effectively decreased levels of secretion of HBsAg and HBeAg than compound 2. Compared with Adefovir Dipivoxil (ADV), compound 1 inhibited the secretion of HBsAg and HBeAg more effective at the same concentration (1.6μg/ml).
【Conclusion】
phyllocoumarin and (-)-epicatechin are two potential anti-HBV agents and show lower cytotoxicity.
联合用药比单一用药更能降低HBV感染者的发病率和死亡率,但长期联合用药导致耐药、副作用限制了抗HBV药物的使用。因此,不断地开发新的抗HBV药物迫在眉睫。目前临床抗HBV药物均为核苷类药物,如拉米夫定、阿德福韦酯、更昔洛韦、法昔洛韦、恩替卡韦和恩曲他滨,等。从天然资源中寻找新的抗HBV药物或先导化合物的研究是国内外新药研制中非常活跃的领域之一,也是我国近期创制新药的一条捷径。在过去20多年里,从天然资源中分离和鉴定出许多抗HBV药物。我国科学家发现北五味子(Schisandra chinensis)粗提物及其多种有效成分作用于多靶点,具有保肝解毒、抗氧化和抗HBV病毒等多方面的作用,并研发成功联苯双脂和双环醇两种具有我国自主知识产权的治疗肝炎药物。通过对狭叶五味子(Schisandra lancifolia)茎叶的化学成分和抗病毒活性研究,我们发现了Lancifodilactone F和Lancifodilactone G等多个具有抗HIV-1活性的化合物。本文对来源于狭叶五味子的化合物phyllocoumarin和(-)-epicatechin对抗HBV活性进行初步研究。
【方法】
筛选研究从狭叶五味子(Schisandra lancifolia)中分离化合物phyllocoumarin (1)和(-)-epicatechin(2)的体外抗HBV活性,以HepG2.2.15细胞为体外研究HBV模型,分别用RPMI-1640完全培养基稀释不同浓度的化合物1和2,与HepG2.2.15细胞共培养3天后收集上清,采用MTT法检测药物对HepG2.2.15细胞的生长影响和ELISA法检测培养上清中HBV HBsAg和HBeAg的含量,评价化合物1和2对HBV复制的影响。
【结果】
化合物1和2具有一定的体外抗HBV活性,其细胞毒性低,CC50均大于200μg/ml。化合物1具有较强的抑制HBsAg和HBeAg分泌作用。阳性对照药物阿德福韦酯也抑制HBV HBsAg和HBeAg分泌,但在相同浓度(1.6μg/ml)下其抑制作用较化合物1弱。
【结论】
phyllocoumarin和(-)-epicatechin能抗HBV活性,且毒性小。
【Objective】
Although the combination of the chemical drugs in the antiviral therapy more effectively reduce morbidity and mortality in HBV infected individuals than the use of single drug, emergence of HBV drugs resistance and side effects for long-term antiretroviral treatment limited the use of these drugs. So continuously development of new anti-HBV drugs appears to be inevitable. Presently, drugs of the anti-HBV therapy used clinically are nucleoside analogues.such as lamivudine, adefovir dipivoxil,ganciclovir, famciclovir and entecavir,emtricitabine. Natural products as the most consistently successful source in drug discovery may offer opportunities for finding anti-HBV drugs or lead compounds. In past 20 years, many natural compounds with anti-HBV activity have been isolated and indentified, In this study, we tested the anti-HBV effects of compounds isolated from Schisandra lancifolia. Scientists in China have found that compouds and many effective ingredients from Schisandra chinensis, with the function of protecting liver and decreasing levels of ALT, antioxidant and anti-HBV activity, tagarted many sites of virus. They have succeeded in developing two anti-HBV drugs, Bifendate and bicyclol. To confirm chemical compositions from Schisandra lancifolia and to evaluate its antiviral therapy,we have found many compouds, such as Lancifodilactone F and Lancifodilactone G, were against HIV-1. Here, we primarily studied that compouds phyllocoumarin (1) and (-)-epicatechin (2) from Schisandra lancifolia were anti-HBV activity.
【Methods】
To evaluate anti-HBV activities of phyllocoumarin (1) and (-)-epicatechin (2) from Schisandra lancifolia in vitro,we used HepG2.2.15 cells as HBV model in vitro. Different concentrations of compounds (1) and compounds (2) were added to the wells with the growth of HepG2.2.15 cells and the supernatant was collected at the third day. The cytotoxic activity on the growth of HepG2.2.15 cells was detected by MTT. ELISA for HBsAg and HBeAg assays were used to evaluate effects of compounds 1 and 2 on HBV replication.
【Result】
Both coupound 1 and compound 2 showed anti-HBV activity and exerted cytotoxicity against HepG2.2.15 cells with CC50>200μg/ml. But coupound 1 more effectively decreased levels of secretion of HBsAg and HBeAg than compound 2. Compared with Adefovir Dipivoxil (ADV), compound 1 inhibited the secretion of HBsAg and HBeAg more effective at the same concentration (1.6μg/ml).
【Conclusion】
phyllocoumarin and (-)-epicatechin are two potential anti-HBV agents and show lower cytotoxicity.
引文
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