Delong R, Stephenson K, Loftus T, Fisher M, Alahari S, Nolting A, Juliano R L
Department of Pharmacology, School of Medicine, University of North Carolina at Chapel Hill 27599, USA.
J Pharm Sci. 1997 Jun;86(6):762-4. doi: 10.1021/js960409f.
This study evaluates polyamidoamine PAMAM "starburst" dendrimers (generation 3, Mr 6909) as a potential delivery vehicle for oligonucleotides. Complexes between dendrimer and phosphorothioate oligonucleotides were observed by agarose gel electrophoresis and were positive, negative, or neutral in charge depending on stoichiometry. Complexes were stable in 50% serum to variations in pH (3, 5, and 10) and ionic strength (0-500 mM). Ultrafiltration and gel filtration characterization indicated that the dendrimer:oligonucleotide complexes were primarily < 100 kD, although some larger complexes were formed at oligonucleotide excess. Use of dendrimers resulted in a 50-fold enhancement in cell uptake of oligonucleotide as determined by flow cytometry, and enhanced cytosolic and nuclear availability, as shown by confocal microscopy. These data support the further evaluation of dendrimers for oligonucleotide delivery in cell culture and in vivo.
本研究评估了聚酰胺-胺（PAMAM）“星爆”树枝状大分子（第3代，分子量6909）作为寡核苷酸潜在递送载体的性能。通过琼脂糖凝胶电泳观察到树枝状大分子与硫代磷酸酯寡核苷酸之间形成的复合物，其电荷根据化学计量比呈正、负或中性。复合物在50%血清中对pH值（3、5和10）及离子强度（0 - 500 mM）的变化具有稳定性。超滤和凝胶过滤表征表明，树枝状大分子与寡核苷酸的复合物主要小于100 kD，不过在寡核苷酸过量时会形成一些较大的复合物。流式细胞术测定结果显示，使用树枝状大分子可使寡核苷酸的细胞摄取增强50倍，共聚焦显微镜观察结果表明其胞质和核内可用性增强。这些数据支持进一步评估树枝状大分子在细胞培养和体内用于寡核苷酸递送的效果。
