Conjugation to 10 kDa Linear PEG Extends Serum Half-Life and Preserves the Receptor-Binding Ability of mmTRAIL with Minimal Stimulation of PEG-Specific Antibodies
Conjugation to 10 kDa Linear PEG Extends Serum Half-Life and Preserves the Receptor-Binding Ability of mmTRAIL with Minimal Stimulation of PEG-Specific Antibodies
复制标题
与 10 kDa 线性 PEG 缀合可延长血清半衰期并保留 mmTRAIL 的受体结合能力，同时对 PEG 特异性抗体的刺激最小
DOI：
10.1021/acs.molpharmaceut.6b00964
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发表时间：
2017-02-01
影响因子：
4.9
通讯作者：
Lu, Xiaofeng
中科院分区：
文献类型：
--
作者：
Nie, Qianxue;Jia, Dianlong;Lu, Xiaofeng
大多数治疗性蛋白质的体内效力较差可能是由于其血清半衰期较短。聚乙二醇化是一种行之有效的方法，并已被临床证明可以改善药代动力学。 mmTRAIL在多种肿瘤细胞中表现出超细胞毒性，但其在小鼠体内的血清半衰期小于10分钟。在此，mmTRAIL-5K、mmTRAIL-10K 和 mmTRAIL-20K 分别通过 mmTRAIL 与 5、10 或 20 kDa mPEG 的 N 末端特异性聚乙二醇化产生。 mmTRAIL-5K、mmTRAIL-10K和mmTRAIL-20K的粒径分别为9.09±2.76、12.62±4.05和15.68±4.95 nm，均高于肾清除阈值（接近7 nm）。因此，mmTRAIL-5K 的血清半衰期为 30 分钟，仅比 mmTRAIL 长 3 倍。然而，mmTRAIL-10K 和 mmTRAIL-20K 均表现出相似的血清半衰期，范围为 350 至 400 分钟，表明 10 或 20 kDa mPEG 的聚乙二醇化显着改善了 mmTRAIL 的药代动力学。然而，mmTRAIL-20K 的死亡受体结合减少了 5 至 8 倍，导致细胞毒性减少了 3 倍。此外，重复施用 mmTRAIL-20K 在大鼠中引发了 mPEG 特异性 IgG 和 IgM 抗体反应。相反，mmTRAIL-10K的受体结合和细胞毒性与mmTRAIL相似。重复施用mmTRAIL-10K并没有明显刺激大鼠和恒河猴的mPEG特异性抗体反应。在三种聚乙二醇化 mmTRAIL 类似物中，mmTRAIL-10K 对携带人类肿瘤异种移植物的小鼠发挥最大的肿瘤抑制作用。这些结果表明，mmTRAIL 与 10 kDa mPEG 的缀合比与 5 或 20 kDa mPEG 的缀合更能增强抗肿瘤效果。
The poor in vivo potencies of most therapeutic proteins might be attributed to their short serum half-lives. PEGylation is a well-established method and has been clinically proven to improve pharmacokinetics. mmTRAIL exhibited supercytotoxicity in a variety of tumor cells, but its serum half-life was less than 10 min in mice. Here, mmTRAIL-5K, mmTRAIL-10K, and mmTRAIL-20K were produced by N-terminus-specific PEGylation of mmTRAIL with 5, 10, or 20 kDa mPEG, respectively. The particle sizes of mmTRAIL-5K, mmTRAIL-10K, and mmTRAIL-20K were 9.09 +/- 2.76, 12.62 +/- 4.05, and 15.68 +/- 4.95 nm, which were higher than the threshold (similar to 7 nm) of renal clearance. Accordingly, mmTRAIL-5K exhibited a serum half-life of 30 min only 3 times longer than that of mmTRAIL. However, both mmTRAIL-10K and mmTRAIL-20K exhibited similar serum half-lives ranging from 350 to 400 min, indicating that PEGylation with 10 or 20 kDa mPEG significantly improved the pharmacokinetics of mmTRAIL. However, death receptor binding of mmTRAIL-20K was reduced 5- to 8-fold, resulting in a 3-fold reduction of cytotoxicity. Additionally, repeated administration of mmTRAIL-20K elicited both mPEG-specific IgG and IgM antibody responses in rats. In contrast, the receptor binding and cytotoxicity of mmTRAIL-10K were similar to those of mmTRAIL. Repeated administration of mmTRAIL-10K did not obviously stimulate mPEG-specific antibody responses in rats and rhesus monkeys. Of the three PEGylated mmTRAIL analogues, mmTRAIL-10K exerted the greatest tumor suppression in mice bearing human tumor xenografts. These results demonstrated that conjugation of mmTRAIL to 10 kDa mPEG was better than that to 5 or 20 kDa mPEG for enhancing antitumor effects.