Acetylation and Amination Protect Angiotensin 1-7 from Physiological Hydrolyzation and Therefore Increases Its Antitumor Effects on Lung Cancer
Acetylation and Amination Protect Angiotensin 1-7 from Physiological Hydrolyzation and Therefore Increases Its Antitumor Effects on Lung Cancer
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DOI：
10.1021/acs.molpharmaceut.8b00181
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发表时间：
2018-06-01
影响因子：
4.9
通讯作者：
Li，Meng
中科院分区：
文献类型：
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作者：
Ma，Xiaowen;Pang，Zhijun;Li，Meng
最近报道的血管紧张素1-7（Ang-（1-7））对多种癌症的抑制作用表明其作为原发性和转移性癌症的治疗剂的潜在用途。然而，其在循环中极短的半衰期极大地损害了其潜在的应用。本文报道了一种血管紧张素-（1-7）类似物肽，其氨基和羧基末端分别经乙酰化和胺化保护。使用高效液相色谱法（HPLC）测定所得类似物Ang-AA的体外和体内降解。同时，进行小RNA干扰和竞争研究以评估Ang-AA与细胞表面Mas受体结合的特异性能力。采用CCK-8细胞计数法、伤口愈合法和Boyden小室法研究Ang-AA对A549细胞的抑制作用。最后，使用动物成像系统和存活观察来观察Ang-AA和紫杉醇（PTX）对小鼠A549异种移植物的协同抑制作用。评价Ang-AA对小鼠的毒性。结果表明，乙酰化和胺化在体内外均能显著抑制Ang-（1-7）的水解。Ang-（1-7）在大鼠体内的半衰期由2.4 ± 0.6min延长至238.7 ± 61.3min（P< 0.001）。Ang-AA对A549细胞增殖、迁移和侵袭的抑制作用明显强于Ang-（1-7）。Ang-AA和PTX的组合对A549异种移植物表现出比Ang-（1-7）和PTX的组合显著更大的协同抑制作用。Ang-AA对小鼠无明显毒性。我们的研究结果表明，乙酰化和胺化是一种简单而有效的方法来生产作为生物活性肽的Ang-（1-7）。
The recently reported inhibitory effects of angiotensin 1–7 (Ang-(1–7)) on various cancers indicate its potential use as a therapeutic agent for primary and metastatic cancers. However, its extremely short half-life in the circulation greatly compromises its potential applications. Here, we reported an Ang-(1–7) analogue peptide with the amino and carboxy termini protected by acetylation and amination. The in vitro and in vivo degradation of the resulting analogue, Ang-AA, were determined using high-performance liquid chromatography (HPLC). At the same time, small RNA interference and competition studies were performed to evaluate the specific capacity of Ang-AA to bind to the cell surface Mas receptor. Cell Counting Kit-8 (CCK8), wound-healing, and Boyden chamber assays were performed to investigate the inhibitory effects of Ang-AA on A549 cells. Finally, the synergistic inhibitory effects of Ang-AA and paclitaxel (PTX) on A549 xenografts in mice were observed using animal imaging systems and survival observations. The toxicity of Ang-AA in mice was evaluated. Our results showed that acetylation and amination significantly inhibited the hydrolyzation of Ang-(1–7) in vitro and in vivo. The half-life of Ang-(1–7) in rats was prolonged from 2.4 ± 0.6 min to 238.7 ± 61.3 min (p< 0.001). The specific binding of Ang-AA to the Mas receptor was well preserved, and Ang-AA exerted significantly greater inhibitory effects on the proliferation, migration, and invasion of A549 cells than Ang-(1–7). The combination of Ang-AA and PTX exhibited a significantly greater synergistic inhibitory effect on A549 xenografts than the combination of Ang-(1–7) and PTX. Ang-AA did not display obvious toxicity in mice. Our findings indicate acetylation and amination is a simple and effective method for producing Ang-(1–7) as a bioactive peptide.