Institut des Sciences et Ingénierie Chimiques, École Polytechnique Fédérale de Lausanne (EPFL), CH-1015 Lausanne, Switzerland.
Early Chemical Development, Pharmaceutical Science, R&D, AstraZeneca, SE-431 83 Mölndal, Sweden.
J Am Chem Soc. 2022 Apr 20;144(15):6734-6741. doi: 10.1021/jacs.1c12442. Epub 2022 Apr 6.
The determination of intracellular drug concentrations can provide a better understanding of the drug function and efficacy. Ideally, this should be performed nondestructively, with no modification of either the drug or the target, and with the capability to detect low amounts of the molecule of interest, in many cases in the μM to nM range (pmol to fmol per million cells). Unfortunately, it is currently challenging to have an experimental technique that provides direct quantitative measurements of intracellular drug concentrations that simultaneously satisfies these requirements. Here, we show that magic-angle spinning dynamic nuclear polarization (MAS DNP) can be used to fulfill these requirements. We apply a quantitative N MAS DNP approach in combination with N labeling to quantify the intracellular amount of the drug [N]CHIR-98014, an activator of the Wingless and Int-1 signaling pathway, determining intracellular drug amounts in the range of tens to hundreds of picomoles per million cells. This is, to our knowledge, the first time that MAS DNP has been used to successfully estimate intracellular drug amounts.
细胞内药物浓度的测定可以更好地了解药物的功能和疗效。理想情况下，这应该是非破坏性的，既不改变药物也不改变靶标，并且能够检测到低浓度的感兴趣分子，在许多情况下，其浓度范围为 μM 到 nM（pmol 到 fmol 每百万个细胞）。不幸的是，目前很难有一种实验技术能够直接定量测量同时满足这些要求的细胞内药物浓度。在这里，我们表明，魔角旋转动态核极化（MAS DNP）可用于满足这些要求。我们应用定量 N MAS DNP 方法结合 N 标记来定量测定药物[N]CHIR-98014 的细胞内含量，[N]CHIR-98014 是 Wnt 和 Int-1 信号通路的激活剂，可确定细胞内药物含量在数十到数百皮摩尔每百万个细胞的范围内。据我们所知，这是首次成功使用 MAS DNP 来估计细胞内药物含量。
