Telomere signaling and metabolic dysfunction are hallmarks of cell aging. New agents targeting these processes might provide therapeutic opportunities, including chemoprevention strategies against cancer predisposition. We report identification and characterization of a pyrazolopyrimidine compound series identified from screens focused on cell immortality and whose targets are glycolytic kinase PGK1 and oxidative stress sensor DJ1. We performed structure–activity studies on the series to develop a photoaffinity probe to deconvolute the cellular targets. In vitro binding and structural analyses confirmed these targets, suggesting that PGK1/DJ1 interact, which we confirmed by immunoprecipitation. Glucose homeostasis and oxidative stress are linked to telomere signaling and exemplar compound CRT0063465 blocked hypoglycemic telomere shortening. Intriguingly, PGK1 and DJ1 bind to TRF2 and telomeric DNA. Compound treatment modulates these interactions and also affects Shelterin complex composition, while conferring cellular protection from cytotoxicity due to bleomycin and desferroxamine. These results demonstrate therapeutic potential of the compound series.

端粒信号传导和代谢功能障碍是细胞衰老的标志。针对这些过程的新药物可能会提供治疗机会，包括针对癌症易感性的化学预防策略。我们报告了从细胞永生性筛选中鉴定出的吡唑并嘧啶化合物系列的鉴定和表征，其靶标是糖酵解激酶 PGK1 和氧化应激传感器 DJ1。我们对该系列进行了结构-活性研究，以开发光亲和探针来解卷积细胞靶标。体外结合和结构分析证实了这些靶标，表明 PGK1/DJ1 相互作用，我们通过免疫沉淀证实了这一点。葡萄糖稳态和氧化应激与端粒信号传导有关，示例化合物 CRT0063465 可以阻断低血糖端粒缩短。有趣的是，PGK1 和 DJ1 与 TRF2 和端粒 DNA 结合。化合物治疗调节这些相互作用，还影响 Shelterin 复合物的组成，同时赋予细胞保护，使其免受博莱霉素和去铁胺引起的细胞毒性。这些结果证明了该化合物系列的治疗潜力。