title: Anti-HIV-1 activity of anti-TAR polyamide nucleic acid conjugated with various membrane transducing peptides.
abstract: The transactivator responsive region (TAR) present in the 5'-NTR of the HIV-1 genome represents a potential target for antiretroviral intervention and a model system for the development of specific inhibitors of RNA-protein interaction. Earlier, we have shown that an anti-TAR polyamide nucleotide analog (PNA(TAR)) conjugated to a membrane transducing (MTD) peptide, transportan, is efficiently taken up by the cells and displays potent antiviral and virucidal activity [B. Chaubey, S. Tripathi, S. Ganguly, D. Harris, R. A. Casale and V. N. Pandey (2005) Virology, 331, 418-428]. In the present communication, we have conjugated five different MTD peptides, penetratin, tat peptide, transportan-27, and two of its truncated derivatives, transportan-21 and transportan-22, to a 16mer PNA targeted to the TAR region of the HIV-1 genome. The individual conjugates were examined for their uptake efficiency as judged by FACScan analysis, uptake kinetics using radiolabeled conjugate, virucidal activity and antiviral efficacy assessed by inhibition of HIV-1 infection/replication. While FACScan analysis revealed concentration-dependent cellular uptake of all the PNA(TAR)-peptide conjugates where uptake of the PNA(TAR)-penetratin conjugate was most efficient as >90% MTD was observed within 1 min at a concentration of 200 nM. The conjugates with penetratin, transportan-21 and tat-peptides were most effective as an anti-HIV virucidal agents with IC50 values in the range of 28-37 nM while IC50 for inhibition of HIV-1 replication was lowest with PNA(TAR)-transportan-27 (0.4 microM) followed by PNA(TAR)-tat (0.72 microM) and PNA(TAR)-penetratin (0.8 microM). These results indicate that anti-HIV-1 PNA conjugated with MTD peptides are not only inhibitory to HIV-1 replication in vitro but are also potent virucidal agents which render HIV-1 virions non-infectious upon brief exposure.
authors.affiliation: Department of Biochemistry and Molecular Biology, UMDNJ-New Jersey Medical School 185 South Orange Avenue, Newark, NJ 07103, USA.
authors.forename: Snehlata
authors.initials: S
authors.lastname: Tripathi
authors.forename: Binay
authors.initials: B
authors.lastname: Chaubey
authors.forename: Sabyasachi
authors.initials: S
authors.lastname: Ganguly
authors.forename: Dylan
authors.initials: D
authors.lastname: Harris
authors.forename: Ralph A
authors.initials: RA
authors.lastname: Casale
authors.forename: Virendra N
authors.initials: VN
authors.lastname: Pandey
mesh_terms: D019380:Anti-HIV Agents / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D016375:Antisense Elements (Genetics) / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D004279:DNA, Viral / Q000378:metabolism; D015696:Gene Products, tat / Q000737:chemistry; D016325:HIV Long Terminal Repeat / Q000187:drug effects*; D015497:HIV-1 / Q000187:drug effects*; D018398:Homeodomain Proteins / Q000737:chemistry; D007700:Kinetics; D009696:Nucleic Acids / Q000737:chemistry / Q000494:pharmacology*; D009711:Nucleotides / Q000737:chemistry / Q000494:pharmacology*; D010446:Peptide Fragments / Q000737:chemistry; D020135:Peptide Nucleic Acids / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000494:pharmacology*; D048348:Reverse Transcription / Q000187:drug effects; D014779:Virus Replication / Q000187:drug effects; D054322:tat Gene Products, Human Immunodeficiency Virus
chemical_list: D019380:Anti-HIV Agents; D016375:Antisense Elements (Genetics); D004279:DNA, Viral; D015696:Gene Products, tat; D018398:Homeodomain Proteins; D009696:Nucleic Acids; D009711:Nucleotides; D010446:Peptide Fragments; D020135:Peptide Nucleic Acids; D010455:Peptides; D011993:Recombinant Fusion Proteins; C497138:penetratin (43-58), human; D054322:tat Gene Products, Human Immunodeficiency Virus; C422951:tat peptide (48-60), Human immunodeficiency virus 1