Cell-penetrating TAT peptide in drug delivery systems: proteolytic stability requirements.
Cell-penetrating TAT peptide in drug delivery systems: proteolytic stability requirements.
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DOI：
10.3109/10717544.2011.567310
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发表时间：
2011-07
期刊：
影响因子：
6
通讯作者：
Torchilin VP
中科院分区：
文献类型：
--
作者：
Koren E;Apte A;Sawant RR;Grunwald J;Torchilin VP
研究了HIV细胞穿透性达特肽（TATp）在TATp修饰的胶束和脂质体表面的稳定性和活性。使用用磷脂酰乙醇胺与“短”PEG的缀合物（PEG-PE）修饰的TATp将TATp部分连接到这些纳米载体的表面。用胰蛋白酶、弹性蛋白酶或胶原酶预孵育后，通过HPLC和荧光检测，使用芴基甲基氯甲酸酯（FMOC）标记研究了这些改性载体表面上的TATp的蛋白水解稳定性。所有测试的酶产生剂量依赖性的TATp切割，如TATp Arg-Arg片段的存在所示。当这些TATp-胶束通过添加较长的PEG-PE嵌段进行修饰时，发生TATp切割的抑制，这表明这些嵌段有效地屏蔽了TATp的蛋白水解。还测试了TATP修饰的载体在EL-4、HeLa和B16-F10细胞中积累的能力。胰蛋白酶处理的TATP-修饰的脂质体和胶束导致减少的摄取和细胞相互作用，通过荧光显微镜和荧光激活的细胞分选技术测量。此外，在胰蛋白酶处理后，观察到负载有多柔比星（Doxil）的TATp修饰的脂质体的细胞毒性降低。总之，空间屏蔽的TATp是必不可少的，以确保其在体内的治疗功能。
The stability and activity of the HIV cell-penetrating TAT peptide (TATp) on the surface of TATp-modified micelles and liposomes in relation to its proteolytic cleavage was investigated. TATp moieties were attached to the surface of these nanocarriers using TATp modified with a conjugate of phosphatidyl ethanolamine with a ‘short’ PEG (PEG-PE). Following pre-incubation with trypsin, elastase, or collagenase, the proteolytic stability of TATp on the surface of these modified carriers was studied by HPLC with fluorescence detection using fluorenylmethyl chloroformate (FMOC) labeling. All tested enzymes produced a dose-dependent cleavage of TATp as shown by the presence of TATp Arg-Arg fragments. Inhibition of TATp cleavage occurred when these TATp-micelles were modified by the addition of longer PEG-PE blocks, indicating an effective shielding of TATp from proteolysis by these blocks. TATp-modified carriers were also tested for their ability to accumulate in EL-4, HeLa, and B16-F10 cells. Trypsin treatment of TATp-modified liposomes and micelles resulted in decreased uptake and cell interaction, as measured by fluorescence microscopy and fluorescence-activated cell sorting techniques. Furthermore, a decrease in the cytotoxicity of TATp-modified liposomes loaded with doxorubicin (Doxil) was observed following trypsin treatment. In conclusion, steric shielding of TATp is essential to ensure its in vivo therapeutic function.
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DOI：
10.2217/nnm.10.30
发表时间：
2010-06
期刊：
Nanomedicine (London, England)
影响因子：
--
作者：
Wang T;D'Souza GG;Bedi D;Fagbohun OA;Potturi LP;Papahadjopoulos-Sternberg B;Petrenko VA;Torchilin VP
通讯作者：
Torchilin VP
影响因子：
3.4
作者：
Hällbrink, M;Florén, A;Langel, Ü
通讯作者：
Langel, Ü
影响因子：
2.9
作者：
Torchilin, Vladimir P.
通讯作者：
Torchilin, Vladimir P.
DOI：
10.1073/pnas.151247498
发表时间：
2001-07-17
影响因子：
11.1
作者：
Torchilin, VP;Rammohan, R;Levchenko, TS
通讯作者：
Levchenko, TS
DOI：
10.1007/978-1-60327-360-2_15
发表时间：
2010
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Kale, Amit A;Torchilin, Vladimir P
通讯作者：
Torchilin, Vladimir P