Anti-Herpes Simplex Virus substances produced by the marine green alga, Dunaliella primolecta

在106种供试微藻中，杜氏藻C-523、D.primolecta C-525、Lyngbya sp. M-9和Lyngbya aerugrneo-coerulea M-12的甲醇提取物对单纯疱疹病毒1型（HSV-1）对Vero细胞的致细胞病变（CPE）有抑制作用。其中绿色石斛（D.primolecta）的抗HSV-1活性最高，10 μ g/ml的<-1>提取物可完全抑制CPE，其活性与相同浓度的阿昔洛韦相当。我们比较了对腺病毒、单纯疱疹病毒-2（HSV-2）、日本脑炎病毒和脊髓灰质炎病毒的抗病毒活性。仅抑制HSV-2的CPE。在HSV吸附和侵入细胞过程中，抗病毒活性明显增强。我们通过延长HSV- 1暴露于提取物的时间来优化抗HSV- 1活性的条件。2小时后，即使是高滴度的HSV-1（10^6 TCID 50/0.1 mL）的CPE也完全失活。利用多种色谱技术从D.primolecta藻体中分离纯化出3种具有抗HSV活性的绿色物质，<-1>经NMR和MS分析，确定其化学结构为脱镁叶绿酸类化合物。

Among 106 microalgae tested, the cytopathic effect (CPE) upon Vero cells of herpes simplex virus, Type 1 (HSV-1) was inhibited by four methanol extracts of Dunaliella bioculata C-523, D.primolecta C-525, Lyngbya sp. M-9 and Lyngbya aerugrneo-coerulea M-12. The green alga, D.primolecta, had the highest anti HSV-1 activity, since 10betagml^<-1> of extract from this alga completely inhibited the CPE.This activity was similar to that of acyclovir at the same concentration. We compared anti-viral activities against adeno virus, herpes simplex virus-2 (HSV-2), Japanese Encephalitis and Polio viruses. Only the CPE of HSV-2 was inhibited. Thus, the factor was specific against HSV.The antiviral activity was apparently excited during HSV adsorption and invasion of the cells. We optimized the conditions for anti HSV- 1 activity by prolonging the exposure of HSV- 1 to the extract. After 2h, the CPE of even a high titer of HSV-1 (10^6 TCID5O/O.1 mL) was completely inactivated. By use of various chromatographic tchniques, three green substances having anti-HSV activity were purified from the algal mass of D.primolecta, and 5 mu g mL^<-1> of this purified substance completely inhibited the CPE.From the analysis of NMR and MS, the chemical structures of the active substances were identified as pheophorbide-like compounds.