Use of the post-insertion technique to insert peptide ligands into pre-formed stealth liposomes with retention of binding activity and cytotoxicity
Use of the post-insertion technique to insert peptide ligands into pre-formed stealth liposomes with retention of binding activity and cytotoxicity
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DOI：
10.1023/a:1014434732752
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发表时间：
2002-03-01
影响因子：
3.7
通讯作者：
Allen, TM
中科院分区：
文献类型：
--
作者：
Moreira, JN;Ishida, T;Allen, TM
目的.如果要进行临床试验，就需要大规模生产配体靶向脂质体的简单方法。我们测试了最近开发的将肽配体插入预先形成的隐形脂质体中的技术。制备G受体拮抗剂靶向脂质体（PLG），并将其与阿霉素（Doxorubicin，阿霉素）负载于PLG中，以人小细胞肺癌H69细胞为模型，研究PLG的细胞结合性和细胞毒性。通过硫醚键将六肽拮抗剂G共价偶联到由马来酰亚胺衍生的聚（乙二醇）（M，2000）二硬脂酰磷脂酰乙醇胺形成的胶束中的聚乙二醇（PEG）末端，然后在一步孵育期间转移到预先形成的脂质体中。为了细胞联系。我们使用放射性标记的脂质体。使用MTT法在体外增殖assay.Results的细胞毒性进行了评价：插入后的肽靶向脂质体的形成的方法导致PLG的生产轴承最大约0.3杯拮抗剂G/μ mol磷脂。相对于非靶向脂质体，这些脂质体与H69细胞的细胞结合增加，并且当装载阿霉素时，它们导致与通过常规偶联技术获得的细胞毒性水平相似的细胞毒性。插入后技术是生产生物活性肽靶向脂质体的一种简单、有效的方法。
Purpose. Simple methods for the large-scale manufacture of ligandtargeted liposomes will be needed if clinical trials are to proceed. We tested a recently developed technology for inserting peptide ligands into preformed Stealth liposomes. Antagonist G-targeted liposomes (PLG) were prepared and loaded with doxorubicin and their cellular association and cytotoxicity were evaluated using the human small cell lung cancer H69 cell line.Methods. The hexapeptide antagonist G was covalently coupled via a thioether bond to the terminus of polyethylene glycol (PEG) in micelles formed from maleimide-derivatized poly(ethylene glycol) (M, 2000) distearoylphosphatidylethanolaminc followed by transfer into preformed liposomes during a one-step incubation. For cellular association. we used radiolabeled liposomes. Cytotoxicity was evaluated using the MTT in vitro proliferation assay.Results: The postinsertion approach to the formation of peptide-targeted liposomes led to the production of PLG bearing a maximum of approximately 0.3 mug antagonist G/mumol phospholipid. These liposomes had increased cellular association to H69 cells relative to nontargeted liposomes and, when loaded with doxorubicin, they resulted in similar levels of cytotoxicity to those obtained by conventional coupling techniques.Conclusions. The postinsertion technique is a simple, effective means for the production of biologically active peptide-targeted liposomes.