Development and In Vivo Application of a Water-Soluble Anticancer Copper Ionophore System Using a Temperature-Sensitive Liposome Formulation
Development and In Vivo Application of a Water-Soluble Anticancer Copper Ionophore System Using a Temperature-Sensitive Liposome Formulation
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DOI：
10.3390/pharmaceutics12050466
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发表时间：
2020-05-01
期刊：
影响因子：
5.4
通讯作者：
Szoboszlai, Norbert
中科院分区：
文献类型：
--
作者：
Gaal, Aniko;Garay, Tamas M.;Szoboszlai, Norbert
制备了含铜和铜离子载体新亚铜灵的脂质体，并对其进行了体外和体内抗癌活性的表征。以1，2-二棕榈酰-sn-甘油基-3-磷脂酰胆碱（DPPC）和氢化大豆磷脂酰胆碱（HSPC）为原料，在铜离子存在下制备了温度敏感的聚乙二醇化脂质体。在70：30的DPPC与HSPC重量比下获得最佳的温度依赖性药物释放。通过pH梯度包封新亚铜灵（以0.2摩尔至1摩尔磷脂施用），同时在脂质体内部使用pH 4.5的未缓冲溶液，在脂质体外部使用pH 7.8的100 mM HEPES缓冲液。铜离子过量存在，产生0.5 mM铜-（新亚铜灵）（2）络合物和0.5 mM游离铜。预热至45 ℃增加了热敏脂质体在短期体外实验中的毒性，而在72小时，所有研究的脂质体表现出类似的体外毒性的铜（II）-新亚铜灵络合物（1：1的比例）。热敏脂质体被发现在减少移植有C26癌细胞的BALB/c小鼠中的肿瘤生长方面更有效，而不管温和的热处理。用[Cu-64] Cu-新亚铜灵脂质体治疗后，通过PET/CT成像验证肿瘤的铜摄取。总之，我们的结果证明了靶向包封在含有过量铜的热敏脂质体中的铜纳米毒素的可行性。
Liposomes containing copper and the copper ionophore neocuproine were prepared and characterized for in vitro and in vivo anticancer activity. Thermosensitive PEGylated liposomes were prepared with different molar ratios of 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC) and hydrogenated soybean phosphatidylcholine (HSPC) in the presence of copper(II) ions. Optimal, temperature dependent drug release was obtained at 70:30 DPPC to HSPC weight ratio. Neocuproine (applied at 0.2 mol to 1 mol phospholipid) was encapsulated through a pH gradient while using unbuffered solution at pH 4.5 inside the liposomes, and 100 mM HEPES buffer pH 7.8 outside the liposomes. Copper ions were present in excess, yielding 0.5 mM copper-(neocuproine)(2) complex and 0.5 mM free copper. Pre-heating to 45 degrees C increased the toxicity of the heat-sensitive liposomes in short-term in vitro experiments, whereas at 72 h all investigated liposomes exhibited similar in vitro toxicity to the copper(II)-neocuproine complex (1:1 ratio). Thermosensitive liposomes were found to be more effective in reducing tumor growth in BALB/c mice engrafted with C26 cancer cells, regardless of the mild hyperthermic treatment. Copper uptake of the tumor was verified by PET/CT imaging following treatment with [Cu-64]Cu-neocuproine liposomes. Taken together, our results demonstrate the feasibility of targeting a copper nanotoxin that was encapsulated in thermosensitive liposomes containing an excess of copper.