In Silico Discovery and Characterization of a Novel Nuclear Transcription Factor‑Y (NF-Y) Inhibitor with Antimitogenic Properties

Nuclear Transcription Factor-Y (NF-Y) is a transcription
factor
that binds CCAAT motifs to regulate gene expression, controlling cell
proliferation, metabolism, and differentiation. NF-Y dysregulation
contributes to diverse pathologies, including cancer, neurological
disorders, cardiovascular disease, and tissue fibrosis. Using in silico molecular docking, we screened a library of eight
million compounds to identify molecules targeting a pocket on the
NF-YB/NF-YC dimer. We identified one compound, designated NFYi5, that
was able to reduce the NF-Y activity. NFYi5 reduced mRNA levels of
NF-Y target genes, while sparing housekeeping gene expression, and
inhibiting cell proliferation. Mechanistic studies revealed that NFYi5
impaired NF-Y–DNA binding and accelerated NF-YA protein degradation,
reducing its half-life from 16.5 ± 1.5 h to 8.5 ± 0.7 h.
Together, these data establish NFYi5 as a small-molecule that can
reduce NF-Y activity and is associated with antimitogenic properties.
This proof-of-concept study demonstrates that NF-Y is pharmacologically
tractable and highlights NFYi5 as a potential lead compound for therapeutic
development in NF-Y-driven diseases.

核转录因子-Y（Nuclear Transcription Factor-Y，缩写NF-Y）是一类可结合CCAAT基序以调控基因表达的转录因子，参与调控细胞增殖、代谢与分化过程。NF-Y表达失调可诱发多种病理状态，涵盖癌症、神经系统疾病、心血管疾病及组织纤维化。本研究采用计算机辅助分子对接（in silico molecular docking）技术，对包含800万种化合物的小分子库进行筛选，以靶向结合NF-YB/NF-YC二聚体上的蛋白结合口袋。最终筛选得到一种命名为NFYi5的化合物，该化合物可有效降低NF-Y的转录活性。NFYi5能够下调NF-Y靶基因的信使核糖核酸（mRNA）水平，且不影响持家基因的表达，同时可抑制细胞增殖。机制研究结果显示，NFYi5可削弱NF-Y与DNA的结合能力，并加速NF-YA蛋白的降解，将其半衰期从16.5±1.5小时缩短至8.5±0.7小时。综上，上述实验数据证实NFYi5是一种可降低NF-Y活性的小分子化合物，具备抗有丝分裂特性。本概念验证研究表明，NF-Y可进行药理学靶向调控，同时提示NFYi5有望成为治疗NF-Y驱动性疾病的潜在先导化合物。