Oxystress inducing antitumor therapeutics via tumor-targeted delivery of PEG-conjugated D-amino acid oxidase
Oxystress inducing antitumor therapeutics via tumor-targeted delivery of PEG-conjugated D-amino acid oxidase
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DOI：
10.1002/ijc.22982
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发表时间：
2008-03-01
影响因子：
6.4
通讯作者：
Maeda, Hiroshi
中科院分区：
文献类型：
--
作者：
Fang, Jun;Deng, Dawei;Maeda, Hiroshi
我们开发了一种产生H2O2的酶——聚乙二醇缀合D-氨基酸氧化酶（PEG-DAO），它通过产生有毒的活性氧，即氧化疗法，表现出强效的抗肿瘤活性，随后利用这一优势，对小鼠肉瘤180实体瘤显示出显着的抗肿瘤效果。的，增强渗透性和保留效果。沿着这个思路，我们在这里报道了使用重组DAO制备PEG-DAO，并通过使用各种肿瘤细胞系和肿瘤模型来观察其抗肿瘤活性。重组DAO（rDAO）是从携带猪DAO表达载体的大肠杆菌BL21（DE3）中获得的，具有高产量（20 mg/l）和高酶活性（5.3 U/mg）。聚乙二醇化 rDAO (PEG-rDAO) 对超声处理、反复冻融、冻干表现出高度稳定性，并表现出优异的体内药代动力学。 PEG-rDAO的分子大小为65 kDa，以纳米颗粒形式存在于水溶液中，平均粒径为119 nm。体外实验显示PEG-rDAO对多种肿瘤细胞具有较强的细胞毒性，而对多种正常细胞的细胞毒性较低。使用2种小鼠肿瘤模型进行体内抗肿瘤治疗，即结肠38肿瘤和Meth A肿瘤模型。静脉注射 PEG-rDAO。在足够的滞后时间后，腹膜内注射 D-脯氨酸（DAO 的底物）。给荷瘤小鼠。因此，成功地实现了肿瘤中 H2O2 的优先生成，从而显着抑制了肿瘤生长，且没有任何明显的副作用。这些发现表明 PEG-rDAO 作为临床开发的新型抗癌策略的潜力。 (c) 2007 年 Wiley-Liss, Inc.
We had developed a H2O2 generating enzyme, polyethylene glycol conjugated D-amino acid oxidase (PEG-DAO), which exhibited potent antitumor activity by generating toxic reactive oxygen species, namely oxidation therapy, subsequently showed remarkable antitumor effect on murine Sarcoma 180 solid tumor, by taking advantage. of, the enhanced permeability and retention effect. Along this line, we report here the preparation of PEG-DAO by use of recombinant DAO and its antitumor activity by using various tumor cell lines and tumor models. Recombinant DAO (rDAO) was obtained from E. coli BL21 (DE3) carrying the porcine DAO expression vector with high yield (20 mg/l) and high enzyme activity (5.3 U/mg). Pegylated rDAO (PEG-rDAO) showed high stability against sonication, repeated freezing/thawing, lyophilization and exhibited superior in vivo pharmacokinetics. PEG-rDAO had a molecular size of 65 kDa and existed as nanoparticles in aqueous solution with mean particle diameter of 119 nm. In vitro experiments showed strong cytotoxicity of PEG-rDAO against various tumor cells, whereas less cytotoxicity was found against various normal cells. In vivo antitumor treatment was carried out using 2 mice tumor models, namely colon 38 tumor and Meth A tumor model. PEG-rDAO was administered i.v. and after an adequate lag time, D-proline (the substrate of DAO) was injected i.p. to the tumor-bearing mice. Consequently, preferential generation of H2O2 in the tumor was successfully achieved, which resulted in remarkable suppression of tumor growth without any visible side effects. These findings suggest a potential of PEG-rDAO as a novel anticancer strategy toward clinical development. (c) 2007 Wiley-Liss, Inc.