A stabilized peptide ligand for multifunctional glioma targeted drug delivery
A stabilized peptide ligand for multifunctional glioma targeted drug delivery
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用于多功能神经胶质瘤靶向药物输送的稳定肽配体
DOI：
10.1016/j.jconrel.2016.09.035
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发表时间：
2016
影响因子：
10.8
通讯作者：
Lu Weiyue
中科院分区：
文献类型：
--
作者：
Ying Man;Shen Qing;Zhan Changyou;Wei Xiaoli;Gao Jie;Xie Cao;Yao Bingxin;Lu Weiyue
由氨基酸组成的肽配体在体内会发生蛋白水解。当修饰在纳米载体表面时，这些肽配体将容易降解，并且靶向功效显著减弱。设计稳定的肽配体用于靶向给药已经受到越来越多的关注。在这里，我们提出了一个稳定的肽配体的设计形成的头到尾的酰胺键作为一个例子。尽管线性肽A7 R（称为LA 7 R）可以特异性结合在神经胶质瘤细胞、新生血管和神经胶质瘤血管生成拟态（VM）上过表达的血管内皮生长因子受体2（VEGFR 2）和神经纤毛蛋白-1（NRP-1），但由于蛋白水解，LA 7 R的肿瘤归巢能力在体内受到极大损害（e.g.in the serum）。利用计算机辅助肽段设计技术，合成了一个环状A7R（cA 7 R）肽段，并将固相肽段合成与天然化学连接相结合，高产率地合成了cA 7 R。从理论上和实验上评估了cA 7 R与两种受体的结合。在我们的模拟模型中，疏水和离子相互作用主导了LA 7 R与受体的结合。有趣的是，cA 7 R采用与LA 7 R不同的结构，在不影响疏水和离子相互作用的情况下保持了与受体的高结合亲和力。在通过形成酰胺键进行头-尾环化后，cA 7 R在小鼠血清中表现出优异的稳定性。将cA 7 R或LA 7 R偶联在负载阿霉素（DOX）的脂质体（cA 7 R-LS/DOX或LA 7 R-LS/DOX）的表面上。体外细胞实验结果表明，cA 7 R-LS/DOX不仅对胶质瘤细胞有较强的抗增殖作用，而且对VM和HUVEC管的破坏作用也明显优于LA 7 R-LS/DOX和普通脂质体（LS/DOX，无肽偶联）。cA 7 R偶联物在胶质瘤中的脂质体蓄积量显著高于LA 7 R偶联物，而cA 7 R-LS/DOX与其他DOX制剂（游离DOX、LS/DOX和LA 7 R-LS/DOX）相比，可显著抑制皮下肿瘤生长。所设计的环状A7R在体内具有靶向胶质瘤细胞、新生血管和VM的能力。考虑到cA 7 R肽的合成容易、与受体的高结合亲和力和增加的稳定性，在本研究中，基于计算机辅助肽设计通过形成酰胺键来设计头-尾环化肽提供了一种替代方法来鉴定蛋白水解稳定的肽配体。
Peptide ligands consisting ofl-amino acids are subject to proteolysisin vivo. When modified on the surface of nanocarriers, those peptide ligands would readily degrade and the targeting efficacy is significantly attenuated. It has received increasing scrutiny to design stable peptide ligands for targeted drug delivery. Here, we present the design of a stable peptide ligand by the formation of a head-to-tail amide bond as an example. Even though the linearl-peptide A7R (termedLA7R) can bind specifically to vascular endothelial growth factor receptor 2 (VEGFR2) and neuropilin-1 (NRP-1) that are overexpressed on glioma cells, neovasculature and glioma vasculogenic mimicry (VM), the tumor-homing capacity ofLA7R is greatly impairedin vivodue to proteolysis (e.g.in the serum). A cyclic A7R (cA7R) peptide was identified by computer-aided peptide design and synthesized with high yield by combining solid phase peptide synthesis and native chemical ligation. The binding of cA7R to both receptors was theoretically and experimentally assessed. In our simulated model hydrophobic and ionic interactions dominated the binding ofLA7R to receptors. It is very interesting that cA7R adopting a different structure fromLA7R retained high binding affinities to receptors without affecting the hydrophobic and ionic interactions. After head-to-tail cyclization by the formation of an amide bond, cA7R exhibited exceptional stability in mouse serum. Either cA7R orLA7R was conjugated on the surface of doxorubicin (DOX) loaded liposomes (cA7R-LS/DOX orLA7R-LS/DOX). The results ofin vitrocellular assays indicated that cA7R-LS/DOX not only displayed stronger anti-proliferative effect against glioma cells, but also demonstrated to be more efficient in destruction of VM and HUVEC tubes in comparison toLA7R-LS/DOX and plain liposomes (LS/DOX, without peptide conjugation). cA7R conjugation could achieve significantly higher accumulation of liposomes in glioma than didLA7R conjugation, which in turn, cA7R-LS/DOX could substantially suppress subcutaneous tumor growth when compared with other DOX formulations (free DOX, LS/DOX andLA7R-LS/DOX). The designed cyclic A7R exhibited the capability of targeting glioma cells, neovasculature and VM simultaneouslyin vivo. Considering the ease of synthesis, high binding affinity to receptors and increased stability of cA7R peptide in the present study, the design of head-to-tail cyclized peptides by the formation of amide bond based on computer-aided peptide design presents an alternative method to identify proteolytically stable peptide ligands.