Department of Chemistry, Texas A&M University, Box 30012, College Station, TX 77842, USA.
Chem Soc Rev. 2021 Sep 7;50(17):9794-9816. doi: 10.1039/d1cs00017a. Epub 2021 Jul 22.
Fluorescent dyes attached to kinase inhibitors (KIs) can be used to probe kinases in vitro, in cells, and in vivo. Ideal characteristics of the dyes vary with their intended applications. Fluorophores used in vitro may inform on kinase active site environments, hence the dyes used should be small and have minimal impact on modes of binding. These probes may have short wavelength emissions since blue fluorophores are perfectly adequate in this context. Thus, for instance, KI fragments that mimic nucleobases may be modified to be fluorescent with minimal perturbation to the kinase inhibitor structure. However, progressively larger dyes, that emit at longer wavelengths, are required for cellular and in vivo work. In cells, it is necessary to have emissions above autofluorescence of biomolecules, and near infrared dyes are needed to enable excitation and observation through tissue in vivo. This review is organized to describe probes intended for applications in vitro, in cells, then in vivo. The readers will observe that the probes featured tend to become larger and responsive to the near infared end of the spectrum as the review progresses. Readers may also be surprised to realize that relatively few dyes have been used for fluorophore-kinase inhibitor conjugates, and the area is open for innovations in the types of fluorophores used.
荧光染料与激酶抑制剂 (KIs) 结合后可用于体外、细胞内和体内探测激酶。理想的染料特性因应用而异。体外使用的荧光团可能会影响激酶活性部位的环境，因此所用的染料应体积小且对结合方式的影响最小。这些探针的发射波长可能较短，因为在这种情况下蓝色荧光团就完全足够了。因此，例如，可对模拟核碱基的 KI 片段进行修饰，使其具有荧光性，而对激酶抑制剂结构的干扰最小。然而，对于细胞内和体内工作，则需要使用更大、发射波长更长的染料。在细胞中，需要有高于生物分子自发荧光的发射，并且需要近红外染料来实现体内组织的激发和观察。本综述按照用于体外、细胞内和体内的探针进行组织，读者将观察到，随着综述的进行，探针的特征往往会变得更大，并对近红外区域有响应。读者可能还会惊讶地发现，用于荧光素-激酶抑制剂偶联物的染料相对较少，该领域在使用的荧光团类型方面有创新的空间。
