Anticancer nucleosides are effective against solid tumors and hematological malignancies, but typically are prone to nucleoside metabolism resistance mechanisms. Using a nucleoside-specific multiplexed high-throughput screening approach, we discovered 4’-ethynyl-2’-deoxycytidine (EdC) as a third-generation anticancer nucleoside prodrug with preferential activity against diffuse large B-cell lymphoma (DLBCL) and acute lymphoblastic leukemia (ALL). EdC requires deoxycytidine kinase (DCK) phosphorylation for its activity and induced replication fork arrest and accumulation of cells in S-phase, indicating it acts as a chain terminator. A 2.1Å co-crystal structure of DCK bound to EdC and UDP reveals how the rigid 4’-alkyne of EdC fits within the active site of DCK. Remarkably, EdC was resistant to cytidine deamination and SAMHD1 metabolism mechanisms and exhibited higher potency against ALL compared to FDA approved nelarabine. Finally, EdC was highly effective against DLBCL tumors and B-ALL in vivo. These data characterize EdC as a pre-clinical nucleoside prodrug candidate for DLBCL and ALL.

中文翻译：

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4'-乙炔基-2'-脱氧胞苷 (EdC) 通过诱导复制应激优先靶向淋巴瘤和白血病亚型

抗癌核苷可有效对抗实体瘤和血液恶性肿瘤，但通常容易出现核苷代谢抵抗机制。使用核苷特异性多重高通量筛选方法，我们发现 4'-乙炔基-2'-脱氧胞苷 (EdC) 作为第三代抗癌核苷前药，对弥漫性大 B 细胞淋巴瘤 (DLBCL) 和急性淋巴细胞瘤具有优先活性白血病（全部）。EdC 需要脱氧胞苷激酶 (DCK) 磷酸化才能发挥其活性，并诱导复制叉停滞和细胞在 S 期的积累，表明它充当链终止剂。DCK 与 EdC 和 UDP 结合的 2.1Å 共晶结构揭示了 EdC 的刚性 4'-炔烃如何适合 DCK 的活性位点。值得注意的是，EdC 对胞苷脱氨和 SAMHD1 代谢机制具有抵抗力，并且与 FDA 批准的奈拉滨相比，表现出更高的抗 ALL 效力。最后，EdC 在体内对 DLBCL 肿瘤和 B-ALL 非常有效。这些数据表明 EdC 是 DLBCL 和 ALL 的临床前核苷前药候选药物。