title: Synthesis, hybridization properties and antiviral activity of lipid-oligodeoxynucleotide conjugates.
abstract: Triethylammonium 1,2-di-O-hexadecyl-rac-glycero-3-H-phosphonate (2) was coupled to the 5' terminus of oligodeoxynucleotides via hydrogen phosphonate solid support DNA synthesis methodology. Duplex DNA oligomers with a single 5'-phospholipid melted at lower temperatures than the corresponding unmodified duplex, but duplexes bearing lipids at each 5' end had higher Tms. In uptake experiments with L929 cells, 8-10 times more lipid-DNA became cell-associated than did unmodified DNA. Unmodified antisense diesters were inactive in a VSV antiviral assay in L929 cells (at up to 200 microM). Attachment of a lipid to the oligomer, however, led to a greater than 90% at 150 microM (greater than 80% at 100 microM) reduction in viral protein synthesis. The antiviral activity depended on the sequence of the oligodeoxynucleotide, but some compounds having little or no base complementarity to the viral target were also effective. Phosphorothioate derivatives reduced viral protein synthesis by 20-30% at 100 microM in the VSV assay. The lipid-DNA compounds were not toxic to the cells at up to 100 microM.
authors.affiliation: Genentech, Inc., Department of Molecular Biology, South San Francisco, CA 94080.
authors.forename: R G
authors.initials: RG
authors.lastname: Shea
authors.forename: J C
authors.initials: JC
authors.lastname: Marsters
authors.forename: N
authors.initials: N
authors.lastname: Bischofberger
mesh_terms: D000998:Antiviral Agents / Q000138:chemical synthesis* / Q000378:metabolism / Q000494:pharmacology; D001483:Base Sequence; D002460:Cell Line; D004247:DNA / Q000378:metabolism / Q000494:pharmacology; D016373:DNA, Antisense; D006358:Hot Temperature; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis* / Q000378:metabolism / Q000494:pharmacology; D010712:Phosphatidic Acids; D010743:Phospholipids; D000644:Quaternary Ammonium Compounds; D013696:Temperature; D014721:Vesicular stomatitis Indiana virus / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000096:biosynthesis
chemical_list: D000998:Antiviral Agents; D016373:DNA, Antisense; D009838:Oligodeoxyribonucleotides; D010712:Phosphatidic Acids; D010743:Phospholipids; D000644:Quaternary Ammonium Compounds; D014764:Viral Proteins; C065033:triethylammonium 1,2-di-O-hexadecyl-glycero-3-H-phosphonate; D004247:DNA