Synthesis and in vitro evaluation of cyclic NGR peptide targeted thermally sensitive liposome.
Synthesis and in vitro evaluation of cyclic NGR peptide targeted thermally sensitive liposome.
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DOI：
10.1016/j.jconrel.2009.12.031
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发表时间：
2010-04-19
影响因子：
10.8
通讯作者：
Dreher, Matthew R.
中科院分区：
文献类型：
--
作者：
Negussie, Ayele H.;Miller, Jenna L.;Reddy, Goutham;Drake, Steven K.;Wood, Bradford J.;Dreher, Matthew R.
环状和线性形式的Asn-Gly-Arg （NGR）基序先前已被证明特异性结合CD13/氨基肽酶N， CD13/氨基肽酶N在肿瘤血管和一些肿瘤细胞中选择性过表达。然而，以前的环状NGR在半胱氨酸残基之间使用了一个易受影响的二硫键桥，由于在脂质体表面相邻肽之间形成了二硫键，这可能会给脂质体靶向带来问题。在这项研究中，我们报道了一种新型环状NGR肽的设计、合成和表征，cKNGRE不含二硫桥。以较高的收率和纯度合成了cKNGRE，并将其附着在俄勒冈绿荧光报告蛋白（cKNGRE- og）和含溶脂温敏脂质体（LTSLs）上。利用核磁共振和质谱技术对cKNGRE进行了鉴定。体外荧光显微镜评估显示ckngr - og与CD13+癌细胞结合并积极摄取，与CD13 -癌细胞结合最小。cKNGRE-OG配体对CD13+癌细胞的亲和力比含有线性NGR的肽高3.6倍。当以多价方式呈现在LTSL表面时，环状和线性NGR对CD13+癌细胞的亲和力同样提高了10倍。ckngre靶向LTSLs在41.3°C时快速释放阿霉素（在<4秒内释放75%），在37°C时释放最小。这些结果证明了合成ckngr靶向温度敏感脂质体的能力，该脂质体缺乏二硫桥，具有足够的结合亲和力，可用于生物应用。
The Asn-Gly-Arg (NGR) motif in both cyclic and linear form has previously been shown to specifically bind to CD13/aminopeptidase N that is selectively overexpressed in tumor vasculature and some tumor cells. However, previous versions of cyclic NGR used a liable disulfide bridge between cysteine residues that may be problematic for liposome targeting due to disulfide bond formation between adjacent peptides on the liposomal surface. In this study, we report the design, synthesis, and characterization of a novel cyclic NGR containing peptide, cKNGRE, which does not contain a disulfide bridge. cKNGRE was synthesized in good yield and purity and attached to the fluorescent reporter Oregon Green (cKNGRE-OG) and lysolipid-containing temperature sensitive liposome (LTSLs). The identity of cKNGRE was verified with NMR and mass spectral techniques. In vitro fluorescence microscopy evaluation of cKNGRE-OG demonstrated binding and active uptake by CD13+ cancer cells and minimal binding to CD13− cancer cells. The cKNGRE-OG ligand displayed 3.6-fold greater affinity for CD13+ cancer cells than a linear NGR containing peptide. Affinity for CD13+ cancer cells was similarly improved 10-fold for both the cyclic and linear NGR when presented in a multivalent fashion on the surface of an LTSL. cKNGRE-targeted LTSLs rapidly released (>75% in <4 sec) doxorubicin at 41.3°C with minimal release at 37°C. These results demonstrate the ability to synthesize a cKNGRE-targeted temperature sensitive liposome that lacks a disulfide bridge and has sufficient binding affinity for biological applications.
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影响因子：
4.7
作者：
Gabizon, A;Horowitz, AT;Zalipsky, S
通讯作者：
Zalipsky, S
DOI：
10.1161/01.atv.0000245807.65714.0b
发表时间：
2006-12-01
影响因子：
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作者：
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通讯作者：
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作者：
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通讯作者：
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影响因子：
82.9
作者：
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通讯作者：
Pasqualini, R