Abe T, Mizuta T, Hatta T, Miyano-Kurosaki N, Fujiwara M, Takai K, Shigeta S, Yokota T, Takaku H
Department of Industrial Chemistry, Chiba Institute of Technology, 2-17-1 Tsudanuma, Narashino, 275-0016, Chiba, Japan.
Eur J Pharm Sci. 2001 Apr;13(1):61-9. doi: 10.1016/s0928-0987(00)00208-6.
The liposomally encapsulated and the free antisense phosphorothioate oligonucleotides (S-ODNs) with four target sites (PB1, PB2, PA, and NP) were tested for their abilities to inhibit virus-induced cytopathogenic effects by a MTT assay using MDCK cells. The liposomally encapsulated S-ODN complementary to the sites of the PB2-AUG initiation codon showed highly inhibitory effects. On the other hand, the inhibitory effect of the liposomally encapsulated S-ODN targeted to PB1 was considerably decreased in comparison with those directed to the PB2 target sites. The liposomally encapsulated antisense phosphorothioate oligonucleotides exhibited higher inhibitory activities than the free oligonucleotides, and showed sequence-specific inhibition, whereas the free antisense phosphorothioate oligonucleotides were observed to inhibit viral absorption to MDCK cells. Therefore, the antiviral effects of S-ODN-PB2-AUG and PA-AUG were examined in a mouse model of influenza virus A infection. Balb/c mice exposed to the influenza virus A (A/PR/8/34) strain at dose of 100 LD(50)s were treated i.v. with various doses (5-40 mg/kg) of liposomally (Tfx-10) encapsulated PB2-AUG or PA-AUG before virus infection and 1 and 3 days postinfection. PB2-AUG oligomer treated i.v. significantly prolonged the mean survival time in days (MDS) and increased the survival rates with a dose-dependent manner. We demonstrate the first successful in vivo antiviral activity of antisense administered i.v. in experimental respiratory tract infections induced with influenza virus A.
通过使用MDCK细胞的MTT试验，测试了具有四个靶位点（PB1、PB2、PA和NP）的脂质体包裹型和游离反义硫代磷酸酯寡核苷酸（S-ODN）抑制病毒诱导的细胞病变效应的能力。与PB2-AUG起始密码子位点互补的脂质体包裹型S-ODN显示出高度抑制作用。另一方面，与针对PB2靶位点的S-ODN相比，靶向PB1的脂质体包裹型S-ODN的抑制作用显著降低。脂质体包裹的反义硫代磷酸酯寡核苷酸比游离寡核苷酸表现出更高的抑制活性，并显示出序列特异性抑制，而游离反义硫代磷酸酯寡核苷酸则被观察到可抑制病毒对MDCK细胞的吸附。因此，在甲型流感病毒感染的小鼠模型中研究了S-ODN-PB2-AUG和PA-AUG 的抗病毒作用。以100个半数致死剂量（LD50）的剂量暴露于甲型流感病毒（A/PR/8/34）株的Balb/c小鼠，在病毒感染前以及感染后1天和3天，通过静脉注射给予不同剂量（5-40 mg/kg）的脂质体（Tfx-10）包裹的PB2-AUG或PA-AUG。静脉注射PB2-AUG寡聚物可显著延长平均存活天数（MDS），并以剂量依赖方式提高存活率。我们首次证明了静脉注射反义药物在甲型流感病毒诱导的实验性呼吸道感染中具有成功的体内抗病毒活性。
