抗结核活性化合物HY-152E是本实验室前期获得的具有良好抗结核活性并拥有授权专利（ZL201210088290.0）的小分子化合物（最低抑菌浓度≤0.09 μg/mL）。为深入探索HY-152E的抗结核机制，本研究利用药物亲和反应靶标稳定性（drug affinity responsive target stability，DARTS）技术并结合蛋白质谱技术，分析可能与HY-152E相互作用的结核分枝杆菌潜在靶标蛋白。将结核分枝杆菌H37Ra的菌体蛋白裂解液与HY-152E共同孵育互作，用不同浓度的链霉菌蛋白酶消化30、45、60 min后，十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（sodium dodecyl sulfate-polyacrylamide gel electrophoresis，SDS-PAGE）分离并比较与HY-152E互作前后菌体蛋白耐受蛋白酶消化的差异条带，分别在相对分子质量70 000和45 000~55 000处观察到差异蛋白条带。利用蛋白质谱技术分析差异条带的蛋白信息，共获得86个蛋白信息。结合结核分枝杆菌数据库及蛋白功能信息，最终筛选到9个蛋白可能是HY-152E的抗结核作用潜在靶标。这些潜在靶点的确定，为后续研究HY-152E的抗结核分子机制奠定了基础。

The compound HY-152E with anti-tuberculosis activity is a small molecule with good anti-tuberculosis activity (MIC≤0.09 μg/mL) in our previously observations (Patent No. ZL201210088290.0). In order to identify the anti-tuberculosis action model of compound HY-152E, the potential direct-binding target of Mycobacterium tuberculosis, which may interact with the compound HY-152E, is analyzed by drug affinity responsive target stability (DARTS) technique and mass spectrometry. DARTS is a general methodology for identifying and studying protein-ligand interactions. The technique is based on the principle that when a small molecule compound binds to a protein, the interaction stabilizes the target protein’s structure such that it becomes protease resistant. Compound HY-152E was added into protein lysates of Mycobacterium tuberculosis H37Ra, and then the samples with or without compound HY-152E were digested using streptomycin protease with different concentrations for 30, 40 or 60 min. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) results showed that there were two differential bands at 70 000 and 45 000-55 000, respectively in different proteolysis samples. The protein information of the two different bands was analyzed using mass spectrometry, and 86 proteins were identified. Nine potential protein targets of compound HY-152E against tuberculosis were screened out after information searching and functional analysis with tuberculosis database. The determination of these potential targets lays the foundation for the further research of anti-tuberculosis molecular mechanism of compound HY-152E.