Riha Isabella A, Campos Miguel A, Jin Xiaokang, Wang Fiona Y, Zhang Chenlu, Dunne Sara F, Cravatt Benjamin F, Zhang Xiaoyu
Department of Chemistry, Northwestern University Evanston IL 60208 USA
Chemistry of Life Processes Institute, Northwestern University Evanston IL 60208 USA.
RSC Med Chem. 2024 Dec 6;16(2):892-906. doi: 10.1039/d4md00681j. eCollection 2025 Feb 19.
Traditional small molecule drugs often target protein activity directly, but challenges arise when proteins lack suitable functional sites. An alternative approach is targeted protein degradation (TPD), which directs proteins to cellular machinery for proteolytic degradation. Recent studies have identified additional E3 ligases suitable for TPD, expanding the potential of this approach. Among these, DCAF16 has shown promise in facilitating protein degradation through both PROTAC and molecular glue mechanisms. In this study, we developed a homogeneous time resolved fluorescence (HTRF) assay to discover new DCAF16 binders. Using an in-house electrophile library, we identified two diastereomeric compounds, with one engaging DCAF16 at cysteines C177-179 and another reducing its expression. We demonstrated that the compound covalently engaging DCAF16 can be transformed into a PROTAC capable of degrading FKBP12.
传统的小分子药物通常直接靶向蛋白质活性，但当蛋白质缺乏合适的功能位点时就会出现挑战。一种替代方法是靶向蛋白质降解（TPD），它将蛋白质导向细胞机制进行蛋白水解降解。最近的研究已经鉴定出了其他适用于TPD的E3连接酶，扩展了这种方法的潜力。其中，DCAF16已显示出通过PROTAC和分子胶机制促进蛋白质降解的前景。在本研究中，我们开发了一种均相时间分辨荧光（HTRF）测定法来发现新的DCAF16结合剂。使用内部亲电试剂库，我们鉴定出两种非对映异构体化合物，一种在半胱氨酸C177 - 179处与DCAF16结合，另一种降低其表达。我们证明了与DCAF16共价结合的化合物可以转化为能够降解FKBP12的PROTAC。
