Delivery of G3139 using releasable PEG-linkers: Impact on pharmacokinetic profile and anti-tumor efficacy
Delivery of G3139 using releasable PEG-linkers: Impact on pharmacokinetic profile and anti-tumor efficacy
复制标题
DOI：
10.1016/j.jconrel.2006.12.021
复制
发表时间：
2007-06-01
影响因子：
10.8
通讯作者：
Stein, C. A.
中科院分区：
文献类型：
--
作者：
Zhao, Hong;Peng, Ping;Stein, C. A.
为了克服酶降解和短血浆半衰期的问题，这可能限制反义寡核苷酸的递送，以及CpG基序的潜在免疫刺激作用，我们利用了聚乙二醇（PEG）技术，该技术采用了各种可释放的接头（rPEG）。具有与G3139（化合物1）相同序列的抗Bcl-2 5 ′-氨基烷基寡核苷酸的5 ′-20 kDa-PEG化不改变其与肝素结合蛋白bFGF的结合，也不改变细胞色素c从用缀合物处理的分离的线粒体的释放。然而，在体外518 A2黑色素瘤细胞中，PEG化导致细胞摄取大大减少。与此形成鲜明对比的是，I的PEG化导致体内药代动力学特征显著改善，半衰期（t（1/2））延长，血浆浓度增加，血浆浓度-时间曲线下面积（AUC）增加。在体内黑色素瘤518 A2异种移植小鼠模型中，用5 '-20 kDa-PEG-1或1处理显示出相似的肿瘤生长抑制。此外，在体外小鼠脾细胞培养系统中，通过可释放的接头将PEG部分连接到I上，可消除G3139所观察到的免疫刺激反应。我们的研究结果证明了聚乙二醇化寡核苷酸在体内使用的潜力，并指出了寡核苷酸活性的体外和体内模型之间的深刻差异。(C)2007 Elsevier B. V.保留所有权利。
In order to overcome the problems of enzymatic degradation and short plasma half life, which can limit the delivery of antisense oligonucleotides, and the potential immuno-stimulatory effects of CpG motifs, we utilized a polyethylene glycol (PEG) technology that employed various releasable linkers (rPEG). 5 '-20kDa-PEGylation of an anti-Bcl-2 5 '-aminoalkyl-oligonucteotide with the same sequence as G3139 (Compound 1) did not alter its binding to the heparin-binding protein bFGF, nor the release of cytochrome c from isolated mitochondria treated with the conjugates. However, in 518A2 melanoma cells in vitro, PEGylation resulted in greatly diminished cellular uptake. In striking contrast, PEGylation of I resulted in dramatically improved pharmacokinetic profiles in vivo, with a prolonged half-life (t(1/2)), increased plasma concentration, and increased area under the plasma concentration-time curve (AUC). In an in vivo melanoma 518A2 xenograft mouse model, treatment with either 5 '-20kDa-PEG-1 or 1 demonstrated similar tumor growth inhibition. Furthermore, in an in vitro mouse splenocyte culture system, attachment of a PEG moiety to I through releasable linkers abolished the immunostimulatory response that was observed for G3139. Our results demonstrate the potential of the in vivo use of PEGylated oligonucleotides, and point out the profound differences between in vitro and in vivo models of oligonucleotide activity. (C) 2007 Elsevier B.V. All rights reserved.