Anionic Amino Acid Dendrimer-Trastuzumab Conjugates for Specific Internalization in HER2-Positive Cancer Cells
Anionic Amino Acid Dendrimer-Trastuzumab Conjugates for Specific Internalization in HER2-Positive Cancer Cells
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DOI：
10.1021/mp100105c
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发表时间：
2010-07-01
影响因子：
4.9
通讯作者：
Hashida, Mitsuru
中科院分区：
文献类型：
--
作者：
Miyano, Takuya;Wijagkanalan, Wassana;Hashida, Mitsuru
曲妥珠单抗是一种抗人表皮生长因子受体2（HER 2）的人源化单克隆抗体，为靶向HER 2表达的肿瘤细胞提供了一种有前途的抗癌药物策略。曲妥珠单抗与树枝状聚合物（具有高度官能化表面的重复支化聚合物）的缀合可以增强药物负载能力。然而，典型的树枝状聚合物，如阳离子聚酰胺-胺树枝状聚合物已表现出非特异性细胞毒性。在本研究中，我们开发了一种新的生物相容性氨基酸树枝状大分子与潜在的毒性较低的表面修饰的第六代赖氨酸树枝状大分子与谷氨酸（KG 6 E）。合成的KG 6 E显示出约5-6 nm的良好控制的粒度，具有低的多聚性和可忽略的细胞毒性的负表面电位。接下来，与游离曲妥珠单抗和KG 6 E树枝状聚合物相比，在HER 2阳性（SKBR 3）和阴性（MCF 7）人乳腺癌细胞系中评价荧光标记的KG 6 E-曲妥珠单抗缀合物的靶向效率。KG 6 E-曲妥珠单抗偶联物以剂量依赖性方式特异性结合SKBR 3细胞，与MCF 7细胞的结合亲和力较低。此外，缀合物在SKBR 3细胞中显著内化，然后运输至溶酶体。这些结果表明KG 6 E-曲妥珠单抗缀合物作为HER 2靶向载体用于癌症治疗的治疗和诊断方法的潜力。
Trastuzumab, a humanized monoclonal antibody against human epidermal growth factor receptor 2 (HER2), offers a promising strategy of anticancer drug targeting to HER2-expressing cancer cells. Conjugation of trastuzumab to dendrimers, repeatedly branched polymers with a highly functionalized surface, can enhance the drug loading capacity. However, typical dendrimers such as cationic polyamidoamine dendrimers have exhibited a nonspecific cytotoxicity. In the present study, we developed a novel biocompatible amino acid dendrimer with potentially less toxicity by surface modification of the sixth generation lysine dendrimer with glutamate (KG6E). The synthesized KG6E showed a well-controlled particle size around 5-6 nm with low polydispersibility and negative surface potentials for negligible cytotoxicity. Next, the targeting efficiency of the fluorescent-labeled KG6E-trastuzumab conjugate was evaluated in HER2-positive (SKBR3) and -negative (MCF7) human breast cancer cell lines compared to free trastuzumab and KG6E dendrimers. The KG6E-trastuzumab conjugate was specifically bound to SKBR3 cells in a dose-dependent manner with low binding affinity to MCF7 cells. Furthermore, the conjugate was significantly internalized in SKBR3 cells and then trafficked to lysosomes. These results indicate the potential of KG6E-trastuzumab conjugates as HER2-targeting carriers for therapeutic and diagnostic approaches to cancer therapy.