N-terminally PEGylated human interferon-β-1a with improved pharmacokinetic properties and in vivo efficacy in a melanoma angiogenesis model
N-terminally PEGylated human interferon-β-1a with improved pharmacokinetic properties and in vivo efficacy in a melanoma angiogenesis model
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DOI：
10.1021/bc050237q
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发表时间：
2006-01-01
影响因子：
4.7
通讯作者：
Pepinsky, RB
中科院分区：
文献类型：
--
作者：
Baker, DP;Lin, EY;Pepinsky, RB
ifn - α的聚乙二醇化已被成功地用于改善药物的药代动力学性质和疗效。为了制备适合体内检测的聚乙二醇形式的人干扰素- β -1a (ifn - β -1a)，我们合成了20 kDa的mpeg - o -2-甲基丙醛，并用其修饰细胞因子的n端a-氨基。聚乙二醇化蛋白在大鼠体内静脉给药后保留了与未修饰蛋白相似的50%的活性，并显着改善了药代动力学特性。在稳定状态下的间隙和分布体积分别减少了约30倍和约4倍，从而导致系统暴露的显着增加（由曲线下面积确定）。聚乙二醇化蛋白的消除半衰期比未修饰的蛋白大13倍。在裸胸纯合（nu/nu）小鼠中，检测了未修饰和聚乙二醇化蛋白抑制进入人SK-MEL-1黑色素瘤周围的径向定向血管形成的能力。在一项单剂量比较研究中，1 X 10(6)单位未修饰的ifn - β -1a导致血管数量减少29%，而1 X 10(6)单位聚乙二醇化的ifn - β -1a导致血管数量减少58%。与对照处理小鼠相比，两种处理均导致平均血管数显著减少，与未修饰的ifn - β -1a处理小鼠相比，聚乙二醇化IFNP-1 α处理小鼠的平均血管数显著减少。在一项多剂量与单剂量比较研究中，每天服用1 × 10(6)个单位的未修饰ifn - β -1a，持续9天，血管数量减少51%，而单剂量服用1 × 10(6)个单位的聚乙二醇化蛋白，血管数量减少66%。两种治疗方法均导致平均血管数量与药物处理小鼠相比有统计学意义上的显著减少，与未修饰的ifn - β -1a处理小鼠相比，聚乙二醇化ifnp - β -1a处理小鼠的平均血管数量有统计学意义上的显著减少。因此，改进的修饰蛋白的药代动力学特性转化为提高的功效。由于未经修饰的ifn - β用于治疗多发性硬化症和丙型肝炎病毒感染，该蛋白的聚乙二醇化形式（如20 kDa mpeg - β -2-甲基丙醛修饰的ifn - β -1a）可能作为治疗这些疾病的有用辅助物。此外，聚乙二醇化ifn - β -1a的抗血管生成作用可能用于某些癌症的治疗，无论是单独使用还是与其他抗肿瘤药物联合使用。
PEGylation of IFN-alpha has been used successfully to improve the pharmacokinetic properties and efficacy of the drug. To prepare a PEGylated form of human interferon-beta-1a (IFN-beta-1a) suitable for testing in vivo, we have synthesized 20 kDa mPEG-O-2-methylpropionaldehyde and used it to modify the N-terminal a-amino group of the cytokine. The PEGylated protein retained similar to 50% of the activity of the unmodified protein and had significantly improved phannacokinetic properties following intravenous administration in rats. The clearance and volume of distribution at steady state were reduced similar to 30-fold and similar to 4-fold, respectively, resulting in a significant increase in systemic exposure as determined by the area under the curve. The elimination half-life of the PEGylated protein was similar to 13-fold greater than for the unmodified protein. The unmodified and PEGylated proteins were tested for their ability to inhibit the formation of radially oriented blood vessels entering the periphery of human SK-MEL-1 melanoma tumors in athymic nude homozygous (nu/nu) mice. In a single dose comparison study, administration of 1 X 10(6) units of unmodified IFN-beta-1a resulted in a 29% reduction in vessel number, while I X 10(6) units of PEGylated IFN-beta-1a a resulted in a 58% reduction. Both treatments resulted in statistically significant reductions in mean vessel number as compared to the vehicle (control)-treated mice, with the PEGylated IFNP-1 alpha-treated mice showing a statistically significantly greater reduction in mean vessel number as compared to the unmodified IFN-beta-1a-treated mice. In a multiple versus single dose comparison study, daily administration of I X 10(6) units of unmodified IFN-beta-1a for 9 days resulted in a 51% reduction in vessel number, while a single dose of 1 x 10(6) units of the PEGylated protein resulted in a 66% reduction. Both treatments resulted in statistically significant reductions in mean vessel number as compared to the vehicle-treated mice, with the PEGylated IFNP-beta-1a-treated mice showing a statistically significantly greater reduction in mean vessel number as compared to the unmodified IFN-beta-1a-treated mice. Therefore, the improved pharmacokinetic properties of the modified protein translated into improved efficacy. Since unmodified IFN-beta is used for the treatment of multiple sclerosis and hepatitis C virus infection, a PEGylated form of the protein such as 20 kDa mPEG-beta-2-methylpropionaldehyde-modified IFN-beta-1a may serve as a useful adjunct for the treatment of these diseases. In addition, the antiangiogenic effects of PEGylated IFN-beta-1a may be harnessed for the treatment of certain cancers, either as a sole agent or in combination with other antitumor drugs.