engleski

Use of cyclodextrin as carrier for oligonucleotides

The use of antisense oligonucleotides as tool for modulating gene expression represents a novel strategy for designing drugs for various diseases. In addition to various other parameters, cellular uptake and internalization of oligonucleotides is an important parameter for its effectiveness as antisense agents. We have used cyclodextrin and its analogs as carriers for oligonucleotides to increase cellular uptake. The studies were carried out using ^35S-labeled or fluorescent-labeled phosphorothioate oligonucleotidein human T cell leukemia H9, CEM and Molt-3 cell line. Cellular uptake of oligonucleotide phosphotothioate in presence of cyclodextrin was found to be dose and time dependent. Using various cyclodextrin analogs, e.g. 2-hydroxypropyl beta-cyclodextrin (hpcd), hydroxyethyl beta-cyclodextrin (HECD) and mixture of various hydroxypropyl beta-cyclodextrin (Ebcapsin), increase in oligonucleotide uptake, up to too to three fold in 48 hours, was observed. Cyclodextrin itself was not toxic at the concentration being used. In Order to further increase the uptake we studied oligonucleotides covalently linked with moieties which have strong affinity for cyclodextrin, e.g., adamantane. In comparative studies, oligonucleotide phosphothioate-adamantane conjugate, more increase in uptake was observed in presence of cyclodextrin. These studies suggest that cyclodextrin and its analogs may be a class of promising novel carrier for oligonucleotides and analogs.

cyclodextrin; oligonucleotides

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