Yang Ya-Dong, Zhao Chen, Li Liang-Peng, Lv Yi-Xin, Yang Bei-Bei, Li Xin, Wang Ru, Li Li
State Key Laboratory of Digestive Health, Beijing Key Laboratory of Active Substances Discovery and Druggability Evaluation, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.
Pharmaceuticals (Basel). 2025 Jun 2;18(6):833. doi: 10.3390/ph18060833.
: Avapritinib is an orally bioavailable tyrosine kinase inhibitor and was approved by the FDA in 2020 for gastrointestinal stromal tumor treatments. Although avapritinib is known to be chiral, its stereochemistry was initially established randomly. This study aims to develop a definitive method for determining avapritinib's absolute configuration and propose a universal methodology for stereochemical characterization of flexible chiral drugs. : The absolute configuration of avapritinib was determined through an integrated approach combining chiral resolution, chiroptical spectroscopy and synthetic validation. Enantiomeric separation was achieved via chiral liquid chromatography, followed by comprehensive chiroptical characterization including electronic circular dichroism (ECD), specific optical rotation and optical rotatory dispersion. Conformational analysis and density functional theory (DFT) calculations correlated experimental spectra with theoretical predictions, facilitating definitive configurational assignment. The stereochemical determination were further verified through ECD derivatization and chemical synthesis. Finally, the enantiomers' kinase inhibition profiles against c-KIT D816V were quantitatively assessed. : Two enantiomers of avapritinib were resolved via chiral HPLC and a Chiralpak IG column. Through combined experimental ECD spectra and time-dependent DFT calculations employing the core extraction method, the -isomer was unambiguously determined as configuration. This stereochemical assignment was confirmed by -cyanobenzaldehyde derivatization and de novo synthesis. Biological evaluation revealed ()-(-)-avapritinib exhibited superior c-KIT D816V inhibitory activity compared to its ()-(+)-counterpart, a finding corroborated by molecular docking studies elucidating their differential target interactions. : This study advances avapritinib stereochemical understanding and establishes a definitive protocol for its absolute configuration assignment, serving as a paradigm for flexible chiral drug characterization.
阿伐替尼是一种口服生物可利用的酪氨酸激酶抑制剂，于2020年被美国食品药品监督管理局（FDA）批准用于治疗胃肠道间质瘤。尽管已知阿伐替尼具有手性，但其立体化学最初是随机确定的。本研究旨在开发一种确定阿伐替尼绝对构型的确定性方法，并提出一种用于柔性手性药物立体化学表征的通用方法。：通过结合手性拆分、旋光光谱和合成验证的综合方法确定了阿伐替尼的绝对构型。通过手性液相色谱实现对映体分离，随后进行包括电子圆二色性（ECD）、比旋光度和旋光色散在内的全面旋光表征。构象分析和密度泛函理论（DFT）计算将实验光谱与理论预测相关联，有助于确定构型归属。通过ECD衍生化和化学合成进一步验证了立体化学测定结果。最后，定量评估了对映体对c-KIT D816V的激酶抑制谱。：阿伐替尼的两种对映体通过手性高效液相色谱（HPLC）和Chiralpak IG柱进行拆分。通过结合实验ECD光谱和采用核心提取方法的含时DFT计算，明确确定 -异构体为 构型。这种立体化学归属通过 -氰基苯甲醛衍生化和从头合成得到证实。生物学评价显示，（）-（-）-阿伐替尼对c-KIT D816V的抑制活性优于其（）-（+）-对映体，分子对接研究阐明了它们不同的靶点相互作用，这一发现与此结果相符。：本研究推进了对阿伐替尼立体化学的理解，并建立了其绝对构型归属的确定性方案，为柔性手性药物表征提供了范例。
