Peptide−Oligonucleotide Phosphorothioate Conjugates with Membrane Translocation and Nuclear Localization Properties

Abstract

Eighteen peptide-oligonucleotide phosphorothioate conjugates were prepared in good yield and thoroughly characterized with electrospray ionization mass spectra. When applied to the living cells, conjugates exhibiting membrane translocation and nuclear localization properties displayed efficient intracellular penetration but failed to show any serious antisense effect. Studies on the intracellular distribution of the fluorescein-labeled conjugates revealed their trapping in endosomes.

The role of cell cycle on polyplex‐mediated gene transfer into a retinal pigment epithelial cell line

“…The doubly transfected subclone D407 6-2 cells that express tetracycline-repressive, stably integrated luciferase activity were generated as described in Antopolsky et al [28]. Transfection of CMVβ plasmid into these cells was carried out and analyzed as described above for synchronized cells.…”

“…D407 6-2 cells that stably expressed tetracycline-repressible luciferase [28] were exposed to polymer/pCMVβ complexes at different time points during the cell cycle. The cells were arrested at different phases as described earlier.…”

Pharmacokinetics and Biodistribution of Novel Aptamer Compositions

“…In contrast to antisense oligonucleotides, however, aptamers are in general longer (30)(31)(32)(33)(34)(35)(36)(37)(38)(39)(40) vs. 10-20 nucleotides), possess different types of chemical modifications (sugar, e.g., 2Ј-F, 2Ј-O-Me, 2Ј-NH 2 , vs. backbone modifications), and assume complex tertiary structures that are, in many respects, more similar to the three-dimensional forms of globular proteins than to nucleic acids. Given these considerable differences, the in vivo disposition of aptamers is not readily predictable from antisense results.…”

“…Given these considerable differences, the in vivo disposition of aptamers is not readily predictable from antisense results. Earlier studies involving antisense oligonucleotides have explored the effects of various conjugation chemistries on pharmacokinetics and biodistribution, with the ultimate goal of increasing delivery of antisense molecules to their sites of action inside cells or within certain tissue types in vivo (29)(30)(31)(32)(33). For example, conjugation with cholesterol has been reported to increase the circulation half-life of antisense oligonucleotides, most likely through association with plasma lipoproteins, and to promote hepatic uptake (34).…”

Stepwise Solid-Phase Synthesis of Peptide-Oligonucleotide Conjugates on New Solid Supports

“…Reversed-phase and ionexchange HPLC retention times of the compounds synthesized are given in the Table. The purity of all conjugates, as assessed from reversed-phase HPLC, was higher than 95%. The final characterization was achieved by electrospray-ionization mass spectrometry (ESI-MS), which has proven to be a powerful method for the characterization of peptide-oligonucleotide conjugates [16]. The measured and calculated average molecular masses of the compounds reported here were in excellent agreement (see Table), the difference between the calculated and measured M r being always less than 0.05%.…”