Bi Xinyuan, He Zhicheng, Luo Zhewen, Huang Wensi, Diao Xingxing, Ye Jian
State Key Laboratory of Systems Medicine for Cancer, School of Biomedical Engineering, Shanghai Jiao Tong University Shanghai 200030 P. R. China
Shanghai Institute of Materia Medica, Chinese Academy of Sciences Shanghai 201210 P. R. China.
Chem Sci. 2024 Aug 2;15(34):13998-4008. doi: 10.1039/d4sc02553a.
Bioorthogonal drug molecules are currently gaining prominence for their excellent efficacy, safety and metabolic stability. Pharmacokinetic study is critical for understanding their mechanisms and guiding pharmacotherapy, which is primarily performed with liquid chromatography-mass spectrometry as the gold standard. For broader and more efficient applications in clinics and fundamental research, further advancements are especially desired in cheap and portable instrumentation as well as rapid and tractable pretreatment procedures. Surface-enhanced Raman spectroscopy (SERS) is capable of label-free detection of various molecules based on the spectral signatures with high sensitivity even down to a single-molecule level. But limited by irreproducibility at low concentrations and spectral interference in complex biofluids, SERS hasn't been widely applied for pharmacokinetics, especially in live animals. In this work, we propose a new method to quantify bioorthogonal drug molecules with signatures at the spectral silent region (SR) by the digital colloid-enhanced Raman spectroscopy (dCERS) technique. This method was first validated using 4-mercaptobenzonitrile in a mixture of analogous molecules, exhibiting reliable and specific identification capability based on the unique SR signature and Poisson-determined quantification accuracy. We further developed a single-step serum pretreatment method and successfully profiled the pharmacokinetic behavior of an anticancer drug, erlotinib, from animal studies. In a word, this method, superior in sensitivity, controllable accuracy, minimal background interference and facile pretreatment and measurement, promises diverse applications in fundamental studies and clinical tests of bioorthogonal drug molecules.
生物正交药物分子目前因其卓越的疗效、安全性和代谢稳定性而备受关注。药代动力学研究对于理解其作用机制和指导药物治疗至关重要，该研究主要以液相色谱 - 质谱作为金标准来进行。为了在临床和基础研究中实现更广泛、更高效的应用，尤其需要在廉价便携的仪器设备以及快速简便的预处理程序方面取得进一步进展。表面增强拉曼光谱（SERS）能够基于光谱特征对各种分子进行无标记检测，具有高灵敏度，甚至能达到单分子水平。但由于低浓度下的不可重复性以及复杂生物流体中的光谱干扰，SERS尚未广泛应用于药代动力学研究，尤其是在活体动物中。在这项工作中，我们提出了一种新方法，通过数字胶体增强拉曼光谱（dCERS）技术，利用光谱沉默区域（SR）的特征来定量生物正交药物分子。该方法首先在类似分子混合物中使用4 - 巯基苯甲腈进行验证，基于独特的SR特征和泊松确定的定量准确性，展现出可靠且特异的识别能力。我们进一步开发了一种单步血清预处理方法，并成功从动物研究中描绘了抗癌药物厄洛替尼的药代动力学行为。总之，该方法在灵敏度、可控准确性、最小背景干扰以及简便的预处理和测量方面具有优势，有望在生物正交药物分子的基础研究和临床试验中得到广泛应用。
