EFFECTS OF CHEMICALLY SYNTHESIZED OLIGODEOXYRIBONUCLEOSIDE METHYLPHOSPHONATES ON VESICULAR STOMATITIS VIRUS PROTEIN SYNTHESIS AND INFECTION (NONIONIC ANALOGS, TRANSLATION, OLIGONUCLEOTIDES, ANTIVIRAL AGENTS).

摘要

Four oligodeoxyribonucleoside methylphosphonate sequences of the type, d-Np(Np)(,7)N were synthesized on a solid support. 5'-Protected nucleoside-3'-methylphosphonic imidazolides were used as reaction intermediates. The coupling reaction could be accelerated by the addition of tetrazole. The yields from the coupling reactions were 82-87% per step. The singly charged oligomers were easily purified by DEAE cellulose chromatography. The chainlengths of the oligomers were confirmed after 5'-end labeling with polynucleotide kinase by partial hydrolysis of the methylphosphonate linkages with 1 M aqueous piperidine followed by polyacrylamide gel electrophoresis of the hydrolysate. Pure oligomers could be obtained in seven days in sufficient quantities for moderate scale biochemical and biological experiments.Oligomers I, II, and III specifically inhibited the synthesis of all five viral proteins in infected cells in a concentration dependent manner. The oligomers had no effects on cellular protein synthesis in uninfected cells, nor on cell growth. The oligomer complementary to the poly (rA) trial only had slight effect on VSV protein synthesis. The non-specific inhibition of viral protein synthesis in infected cells may be a consequence of potential partial duplex formation between the oligomers and complementary viral mRNA or the formation of base pairs between the oligomers and plus-stranded viral RNA. These results suggest that these oligomers interact with the viral nucleic acids in infected cells.Oligomers I-III also significantly inhibited VSV production in a manner similar to their effects on VSV protein synthesis. Oligomer IV had no effect. These oligomers therefore may act as antiviral agents.The oligomers were complementary to N, NS, and G VSV nRNAs and to the poly (rA) tail. The oligomers complementary to N (oligomer I) and NS (oligomer II) mRNAs inhibited the translation of VSV but not globin mRNA in a rabbit reticulocyte lysate cell-free system. At 100 (mu)M, oligomer I specifically inhibited N protein synthesis in vitro. In contrast, oligomer II inhibited N as well as NS protein synthesis. This reduced specificity of inhibition may be due to the potential formation of a larger number of partial duplexes between all VSV mRNAs and oligomer II compared to oligomer I. These results indicate that inhibition of viral protein synthesis in vitro occurs as a result of the interaction between the oligomer and the viral mRNA.