Fusion to an albumin-binding domain with a high affinity for albumin extends the circulatory half-life and enhances the in vivo antitumor effects of human TRAIL
Fusion to an albumin-binding domain with a high affinity for albumin extends the circulatory half-life and enhances the in vivo antitumor effects of human TRAIL
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DOI：
10.1016/j.jconrel.2016.03.004
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发表时间：
2016-04-28
影响因子：
10.8
通讯作者：
Lu, Xiaofeng
中科院分区：
文献类型：
--
作者：
Li, Rui;Yang, Hao;Lu, Xiaofeng
重组人肿瘤坏死因子相关凋亡诱导配体（hTRAIL）的药代动力学性能较差，限制了其临床应用。利用内源性白蛋白作为载体是延长循环半衰期的一种有吸引力的方法。在这里，我们通过将蛋白G的白蛋白结合域（ABD）融合到hTRAIL的n或c端，产生了ABD-hTRAIL和hTRAIL-ABD。我们发现ABD-hTRAIL以高亲和力（0.4 +/- 0.18 nM）结合人血清白蛋白（HSA），并形成平均直径（约12 nM）高于肾滤过阈值（约7 nM）的纳米颗粒。ABD-hTRAIL还能结合小鼠血清白蛋白（MSA）；因此，其半衰期是hTRAIL的40-50倍（14.1 +/- 0.87 h vs 0.32 +/- 0.14 h）。注射后8-48 h， ABD-hTRAIL的肿瘤摄取是hTRAIL的6-16倍。因此，ABD-hTRAIL对皮下异种移植物小鼠的肿瘤抑制作用是hTRAIL的3-4倍。此外，ABD-hTRAIL杀死循环肿瘤细胞的时间大约是hTRAIL的8倍。这些结果表明，ABD与n端融合使hTRAIL具有白蛋白结合能力；ABD一旦进入血管，介导与内源性白蛋白结合，从而延长了半衰期，增强了hTRAIL的抗肿瘤作用。然而，hTRAIL-ABD对白蛋白没有高亲和力，因此没有表现出延长循环半衰期和增强抗肿瘤作用。这些结果表明，n端而非c端abd融合是利用内源性白蛋白作为载体增强hTRAIL抗肿瘤作用的有效技术。(C) 2016 Elsevier B.V.版权所有
Clinical applications of recombinant human tumor necrosis factor-related apoptosis-inducing ligand (hTRAIL) have been limited by their poor pharmacokinetics. Using endogenous albumin as a carrier is an attractive approach for circulatory half-life extension. Here, we produced ABD-hTRAIL and hTRAIL-ABD by fusing the albumin-binding domain (ABD) from protein G to the N-or C-terminus of hTRAIL. We found that ABD-hTRAIL bound human serum albumin (HSA) with a high affinity (0.4 +/- 0.18 nM) and formed nanoparticles with an average diameter (similar to 12 nm) above the threshold (similar to 7 nm) of renal filtration. ABD-hTRAIL also bound mouse serum albumin (MSA); thus, its half-life was 40-50-fold greater than that of hTRAIL (14.1 +/- 0.87 h vs 0.32 +/- 0.14 h). Tumor uptake of ABD-hTRAIL 8-48 h post-injection was 6-16-fold that of hTRAIL. Consequently, the tumor suppression of ABD-hTRAIL in mice bearing subcutaneous xenografts was 3-4 times greater than that of hTRAIL. Additionally, the time period during which ABD-hTRAIL could kill circulating tumor cells was approximately 8 times longer than that of hTRAIL. These results demonstrate that ABD fused to the N-terminus endows hTRAIL with albumin binding ability; once it enters the vasculature, ABD mediates binding with endogenous albumin, thus prolonging the half-life and enhancing the antitumor effect of hTRAIL. However, hTRAIL-ABD did not show a high affinity for albumin and therefore did not display the prolonged circulatory half-life and enhanced antitumor effects. These results demonstrate that N-terminal, but not C-terminal, ABD-fusion is an efficient technique for enhancing the antitumor effects of hTRAIL by using endogenous albumin as a carrier. (C) 2016 Elsevier B.V. All rights reserved.