Junghans Monika, Loitsch Stefan M, Steiniger Sebastian C J, Kreuter Jörg, Zimmer Andreas
Institute for Pharmaceutical Technology, Biocenter, Johann Wolfgang Goethe-Universität, Frankfurt am Main, Germany.
Eur J Pharm Biopharm. 2005 Jul;60(2):287-94. doi: 10.1016/j.ejpb.2005.01.006.
In the present study, cationic lipid-peptide-DNA-complexes (LPDs) consisting of AH-Chol-liposomes and protamine-phosphodiester-oligonucleotide-particles (proticles) were introduced as carriers for antisense therapy. The LPDs were physically characterized, and a possible mechanism for adsorption of oligonucleotides (ODNs) was suggested. An increase in stability of ODNs against DNase I and serum nuclease digestion by these carriers was demonstrated. The hydrodynamic diameter increased after incubation with FCS which could be attributed to a protein coating of the particle surface. However, in cell culture medium lower particle sizes of the complexes occurred. In an antisense c-myc in vitro model, the effect of LPDs was tested using U937 cells. The C-MYC level was reduced after treatment of these antisense ODN carrier complexes. Furthermore, no changes in target mRNA concentration of the treated cells was found by reverse transcription and competitive multiplex-PCR.
在本研究中，由AH-胆固醇脂质体和鱼精蛋白-磷酸二酯-寡核苷酸颗粒（proticles）组成的阳离子脂质-肽-DNA复合物（LPDs）被用作反义治疗的载体。对LPDs进行了物理表征，并提出了寡核苷酸（ODNs）吸附的可能机制。证明了这些载体可提高ODNs对DNase I和血清核酸酶消化的稳定性。与胎牛血清（FCS）孵育后，流体动力学直径增加，这可能归因于颗粒表面的蛋白质包被。然而，在细胞培养基中，复合物的粒径较小。在反义c-myc体外模型中，使用U937细胞测试了LPDs的效果。用这些反义ODN载体复合物处理后，C-MYC水平降低。此外，通过逆转录和竞争性多重PCR未发现处理细胞的靶mRNA浓度有变化。
