Luciferin derivatives for enhanced in vitro and in vivo bioluminescence assays
Luciferin derivatives for enhanced in vitro and in vivo bioluminescence assays
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DOI：
10.1021/bi060475o
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发表时间：
2006-09-19
期刊：
影响因子：
2.9
通讯作者：
Contag, Christopher H.
中科院分区：
文献类型：
--
作者：
Shinde, Rajesh;Perkins, Julie;Contag, Christopher H.
体内生物发光成像已成为临床前分子成像的基础技术。这种成像方法是基于发光酶，荧光素酶，它需要特定的底物来产生光。当与人类生物学或疾病的动物模型中的特定生物过程相关联时，酶-底物相互作用成为可以在活体动物中进行无创研究的生物指标。这些动物模型中的信号强度取决于完整器官中活细胞内反应的底物的可用性。因此，底物的生物分布和清除率与最佳成像时间和信号强度直接相关，并最终决定了检测的灵敏度和模型的可预测性。从甲虫（包括萤火虫）中获得的荧光素酶的底物d -荧光素的修饰产生了新的特性，并为改进生物测定提供了机会。为此，我们合成了一种共轭物甘氨酸- d -氨基荧光素，并研究了它与d -氨基荧光素和d -荧光素的相对性质。由于分子结构的差异，这三种底物表现出不同的动力学特性和不同的细胞内积累曲线，从而影响它们在动物体内的生物分布。甘氨酸- d -氨基荧光素的体内循环时间较其他两种底物长。利用这些表现出不同药代动力学和药效学特性的底物，在体外和体内测定荧光素酶的能力将提供灵活性并提高当前的成像能力。
In vivo bioluminescence imaging has become a cornerstone technology for preclinical molecular imaging. This imaging method is based on light-emitting enzymes, luciferases, which require specific substrates for light production. When linked to a specific biological process in an animal model of human biology or disease, the enzyme-substrate interactions become biological indicators that can be studied noninvasively in living animals. Signal intensity in these animal models depends on the availability of the substrate for the reaction within living cells in intact organs. The biodistribution and clearance rates of the substrates are therefore directly related to optimal imaging times and signal intensities and ultimately determine the sensitivity of detection and predictability of the model. Modifications of D-luciferin, the substrate for the luciferases obtained from beetle, including fireflies, result in novel properties and offer opportunities for improved bioassays. For this purpose, we have synthesized a conjugate, glycine-D-aminoluciferin, and investigated its properties relative to those of D-aminoluciferin and D-luciferin. The three substrates exhibited different kinetic properties and different intracellular accumulation profiles due to differences in their molecular structure, which in turn influenced their biodistribution in animals. Glycine-D-aminoluciferin had a longer in vivo circulation time than the other two substrates. The ability to assay luciferase in vitro and in vivo using these substrates, which exhibit different pharmacokinetic and pharmacodynamic properties, will provide flexibility and improve current imaging capabilities.