Antisense therapy of influenza.
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.
Abe
T
T
Department of Industrial Chemistry, Chiba Institute of Technology, 2-17-1 Tsudanuma, Narashino, 275-0016, Chiba, Japan.
Mizuta
T
T
Hatta
T
T
Miyano-Kurosaki
N
N
Fujiwara
M
M
Takai
K
K
Shigeta
S
S
Yokota
T
T
Takaku
H
H
D000818:Animals; D012321:DNA-Directed RNA Polymerases / Q000494:pharmacology / Q000627:therapeutic use*; D009980:Influenza A virus / Q000187:drug effects / Q000472:pathogenicity; D008081:Liposomes; D051379:Mice; D008807:Mice, Inbred BALB C; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D016376:Oligonucleotides, Antisense / Q000494:pharmacology / Q000627:therapeutic use*; D009976:Orthomyxoviridae Infections / Q000188:drug therapy*; D012324:RNA-Dependent RNA Polymerase; D014758:Viral Core Proteins / Q000494:pharmacology / Q000627:therapeutic use*; D014764:Viral Proteins / Q000494:pharmacology / Q000627:therapeutic use*
D008081:Liposomes; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D016376:Oligonucleotides, Antisense; C061290:PA protein, influenza viruses; C057027:PB2 protein, Influenzavirus A; D014758:Viral Core Proteins; D014764:Viral Proteins; C077346:influenza virus polymerase basic protein 1; D012324:RNA-Dependent RNA Polymerase; D012321:DNA-Directed RNA Polymerases