Lu X M, Fischman A J, Jyawook S L, Hendricks K, Tompkins R G, Yarmush M L
Surgical Services, Massachusetts General Hospital, Boston 02114.
J Nucl Med. 1994 Feb;35(2):269-75.
Antisense oligodeoxynucleotides coupled to asialoglycoprotein carrier molecules were evaluated in terms of their ability to accumulate preferentially in the liver and thus potentially serve as an important method to regulate liver gene expression.
Native and asialo-human alpha-1 acid glycoproteins were derivatized with low molecular weight poly(L)lysine and complexed with an antisense DNA (67 mer) complementary to the 5' end of rat serum albumin mRNA. The asialoglycoprotein antisense complex (conjugate) was characterized with respect to size, stability, and anti-sense loading, and the biodistribution of the conjugate was determined for normal rats at 5 min and 1, 6, and 24 hr after intravenous injection. In vivo stability of the anti-sense asialoglycoprotein complex was also evaluated using double-labeled (32P-antisense and 3H-glycoprotein) preparations.
The results of the conjugate characterization studies demonstrated that at least 30% of the anti-sense DNA dissociated from the carrier after 7 min under chromatographic conditions. When the conjugate was incubated with PBS, MEM or MEM plus 10% FBS for 1 hr at 37 degrees C, about 85% of the antisense DNA was dissociated from the carrier. The results of the biodistribution studies showed that the accumulation of the asialo-glycoprotein anti-sense complex in the liver was rapid and greatly exceeded the accumulation of the sialo-glycoprotein antisense analog or antisense alone.
These findings have significant implications for the targeted delivery of therapeutic antisense molecules to the liver.
评估了与去唾液酸糖蛋白载体分子偶联的反义寡脱氧核苷酸优先在肝脏中积累的能力，因此其有可能成为调节肝脏基因表达的重要方法。
天然和去唾液酸人α-1酸性糖蛋白用低分子量聚赖氨酸进行衍生化，并与与大鼠血清白蛋白mRNA 5'端互补的反义DNA（67聚体）复合。对去唾液酸糖蛋白反义复合物（缀合物）的大小、稳定性和反义负载进行了表征，并在静脉注射后5分钟、1小时、6小时和24小时测定了正常大鼠体内缀合物的生物分布。还使用双标记（32P-反义及3H-糖蛋白）制剂评估了反义去唾液酸糖蛋白复合物在体内的稳定性。
缀合物表征研究结果表明，在色谱条件下，7分钟后至少30%的反义DNA从载体上解离。当缀合物在37℃下与PBS、MEM或MEM加10%FBS孵育1小时时，约85%的反义DNA从载体上解离。生物分布研究结果表明，去唾液酸糖蛋白反义复合物在肝脏中的积累迅速，且大大超过了唾液酸糖蛋白反义类似物或单独反义物的积累。
这些发现对于将治疗性反义分子靶向递送至肝脏具有重要意义。
