Abstract：
In this work, we report the identification of novel bromodomain-containing protein 9 (BRD9) binders through a virtual screening based on our developed 3D structure-based pharmacophore model. The in silico workflow here described led to the identification of a promising initial hit (1) featuring the 1-ethyl-1H-pyrazolo[3,4-b]pyridine motif which represented an unexplored chemotype for the development of a new class of BRD9 ligands. The encouraging biophysical results achieved for compound 1 prompted us to explore further tailored structural modification around the C-4 and C-6 positions of the central core. Hence, the design and synthesis of a set of 19 derivatives (2-20) were performed to extensively investigate the chemical space of BRD9 binding site. Among them, four compounds (5, 11, 12, and 19) stood out in biophysical assays as new valuable BRD9 ligands featuring IC50 values in the low-micromolar range. Noteworthy, a promising antiproliferative activity was detected in vitro for compound 5 on HeLa and A375 cancer cell line. The successful combination and application of in silico tools, chemical synthesis, and biological assays allowed to identify novel BRD9 binders and to expand the arsenal of promising chemical entities amenable to the recognition of this important epigenetic target.
摘要：
在这项工作中,我们报告了通过基于我们开发的基于3D结构的药效团模型的虚拟筛选鉴定新型含溴结构域蛋白9(BRD9)结合剂.此处描述的计算机工作流程导致鉴定出具有1-乙基-1H-吡唑并[3,4-b]吡啶基序的有希望的初始命中（1），该基序代表了未开发的化学型，用于开发新一类BRD9配体。化合物1获得的令人鼓舞的生物物理结果促使我们探索围绕中心核的C-4和C-6位置的进一步定制的结构修饰。因此，我们设计和合成了一组19个衍生物(2-20),以广泛研究BRD9结合位点的化学空间.其中,4种化合物(5,11,12和19)在生物物理分析中脱颖而出,作为新的有价值的BRD9配体,其IC50值在低微摩尔范围内.值得注意的是,在体外检测到化合物5对HeLa和A375癌细胞系的有希望的抗增殖活性。silio工具的成功组合和应用，化学合成,和生物测定允许鉴定新的BRD9结合剂，并扩大适合识别这一重要表观遗传靶标的有希望的化学实体的武器库。
