School of Chemistry, Bangor University, Bangor, Gwynedd, LL57 2DG, Wales, United Kingdom.
Langmuir. 2011 Dec 6;27(23):14300-7. doi: 10.1021/la202951p. Epub 2011 Nov 4.
Directed enzyme prodrug therapy is an extensive area of research in cancer chemotherapy. Although very promising, the current directed approaches are still hampered by inefficient enzyme expression and tumor targeting. This work investigates the viability of using metal nanoparticles as a novel delivery vehicle for prodrug-activating enzymes. Using genetically incorporated amino acid sequences, a nitroreductase from E. coli was directly immobilized onto a 50 nm gold colloid, as confirmed by gel electrophoresis, DLS, and UV-vis spectroscopy. The resulting conjugates showed excellent stability in changing proton and sodium chloride environments, including PBS at 37 °C. Remarkably, the immobilized nitroreductase retained more than 99% activity to the CB1954 prodrug without the need for stabilizers. This work provides the foundation for attaching prodrug-activating enzymes to metal nanoparticles for future use in directed enzyme prodrug therapy.
导向酶前药治疗是癌症化疗中一个广泛的研究领域。尽管非常有前途，但目前的导向方法仍然受到酶表达和肿瘤靶向效率低下的限制。本工作研究了将金属纳米粒子用作前药激活酶的新型递送载体的可行性。通过基因整合的氨基酸序列，将来自大肠杆菌的硝基还原酶直接固定在 50nm 的金胶体上，这一点通过凝胶电泳、DLS 和 UV-vis 光谱得到了证实。所得的缀合物在改变质子和氯化钠环境中表现出极好的稳定性，包括 37°C 的 PBS。值得注意的是，固定化的硝基还原酶在不需要稳定剂的情况下对 CB1954 前药保留了超过 99%的活性。这项工作为将前药激活酶附着到金属纳米粒子上以用于未来的导向酶前药治疗奠定了基础。
