title: Improved biological activity of antisense oligonucleotides conjugated to a fusogenic peptide.
abstract: Recently several groups reported a dramatic improvement of reporter gene transfection efficiency using a fusogenic peptide, derived from the Influenza hemagglutinin envelop protein. This peptide changes conformation at acidic pH and destabilizes the endosomal membranes thus resulting in an increased cytoplasmic gene delivery. We describe the use of a similar fusogenic peptide in order to improve the antiviral potency of antisense oligodeoxynucleotides (anti TAT) and oligophosphorothioates (S-dC28) on de novo HIV infected CEM-SS lymphocytes in serum-free medium. We observed as 5 to 10 fold improvement of the anti HIV activities of the phosphodiester antisense oligonucleotides after chemical coupling to the peptide in a one to one ratio by a disulfide or thioether bond. No toxicities were observed at the effective doses (0.1-1 microM). No sequence specificity was obtained and the fusogenic peptide possessed some antiviral activities on its own (IC50: 6 microM). A S-dC28-peptide disulfide linked conjugate and a streptavidin-peptide-biotinylated S-dC28 adduct showed similar activities as the free S-dC28 oligonucleotide (IC50: 0.1-1 nM). As expected, all the compounds were less potent in the presence of serum but the relative contribution of peptide coupling was maintained.
authors.affiliation: Institut de Génétique Moléculaire de Montpellier-UMR CNRS 9942, Montpellier, France.
authors.forename: J P
authors.initials: JP
authors.lastname: Bongartz
authors.forename: A M
authors.initials: AM
authors.lastname: Aubertin
authors.forename: P G
authors.initials: PG
authors.lastname: Milhaud
authors.forename: B
authors.initials: B
authors.lastname: Lebleu
mesh_terms: D000595:Amino Acid Sequence; D000998:Antiviral Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000737:chemistry; D001483:Base Sequence; D002462:Cell Membrane / Q000187:drug effects; D002478:Cells, Cultured; D003432:Cross-Linking Reagents; D016895:Culture Media, Serum-Free; D004337:Drug Carriers; D005347:Fibroblasts / Q000821:virology; D015696:Gene Products, tat; D015497:HIV-1 / Q000187:drug effects* / Q000254:growth & development; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral / Q000737:chemistry*; D008081:Liposomes; D008214:Lymphocytes / Q000821:virology; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D016376:Oligonucleotides, Antisense / Q000737:chemistry / Q000494:pharmacology*; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D019809:Streptavidin; D013873:Thionucleotides / Q000737:chemistry; D054322:tat Gene Products, Human Immunodeficiency Virus
chemical_list: D000998:Antiviral Agents; D001426:Bacterial Proteins; D003432:Cross-Linking Reagents; D016895:Culture Media, Serum-Free; D004337:Drug Carriers; D015696:Gene Products, tat; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral; D008081:Liposomes; D009838:Oligodeoxyribonucleotides; D016376:Oligonucleotides, Antisense; D010455:Peptides; D013873:Thionucleotides; D054322:tat Gene Products, Human Immunodeficiency Virus; C059536:Sd(C)28; D019809:Streptavidin