A cell-based high-throughput screen identifies drugs that cause bleeding disorders by off-targeting the vitamin K cycle
A cell-based high-throughput screen identifies drugs that cause bleeding disorders by off-targeting the vitamin K cycle
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DOI：
10.1182/blood.2019004234
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发表时间：
2020-08-13
期刊：
影响因子：
20.3
通讯作者：
Tie, Jian-Ke
中科院分区：
文献类型：
--
作者：
Chen, Xuejie;Li, Caihong;Tie, Jian-Ke
药物引起的出血性疾病导致大量的发病率和死亡率。引起明显原因的意外出血的抗血栓药物相对容易控制。然而，人们对大多数药物引起的出血性疾病的机制知之甚少，这使得干预变得更加困难。由于大多数出血性疾病与凝血因子功能障碍有关，我们采用了最近建立的基于细胞的测定法来识别影响活性维生素 K 依赖性 (VKD) 凝血因子生物合成的药物，并可能产生不利的脱靶结果。对包含 727 种药物的美国国立卫生研究院 (NIH) 临床保藏中心 (NCC) 库进行了筛选，确定了 9 种药物，其中包括最常用的抗凝药物华法林。与大多数这些药物相关的出血并发症已在临床上有报道，但其致病机制仍不清楚。对抑制 VKD 羧化作用的 9 种热门药物的进一步表征表明，华法林、兰索拉唑和硝唑尼特主要针对维生素 K 环氧化物还原酶（VKOR），而艾地苯醌、氯法齐明和 AM404 主要针对维生素 K 氧化还原循环中的维生素 K 还原酶（VKR）。其他 3 种药物主要影响细胞内维生素 K 的利用率。这些药物使 VKOR 和 VKR 失活的分子机制已得到阐明。基于细胞和动物模型研究的结果表明，靶向 VKOR（而非 VKR）的药物的抗凝作用可以通过服用维生素 K 来挽救。这些发现为药物引起的出血性疾病的预防和管理提供了见解。已建立的基于细胞的高通量筛选方法为鉴定具有抗凝作用的新型维生素 K 拮抗剂提供了强大的工具。
Drug-induced bleeding disorders contribute to substantial morbidity and mortality. Antithrombotic agents that cause unintended bleeding of obvious cause are relatively easy to control. However, the mechanisms of most drug-induced bleeding disorders are poorly understood, which makes intervention more difficult. As most bleeding disorders are associated with the dysfunction of coagulation factors, we adapted our recently estab-lished cell-based assay to identify drugs that affect the biosynthesis of active vitamin K-dependent (VKD) coagulation factors with possible adverse off-target results. The National Institutes of Health (NIH) Clinical Collection (NCC) library containing 727 drugs was screened, and 9 drugs were identified, including the most commonly prescribed anticoagulant warfarin. Bleeding complications associated with most of these drugs have been clinically reported, but the pathogenic mechanisms remain unclear. Further charac-terization of the 9 top-hit drugs on the inhibition of VKD carboxylation suggests that warfarin, lansoprazole, and nitazoxanide mainly target vitamin K epoxide reductase (VKOR), whereas idebenone, clofazimine, and AM404 mainly target vitamin K reductase (VKR) in vitamin K redox cycling. The other 3 drugs mainly affect vitamin K availability within the cells. The molecular mechanisms underlying the inactivation of VKOR and VKR by these drugs are clarified. Results from both cell-based and animal model studies suggest that the anticoagulation effect of drugs that target VKOR, but not VKR, can be rescued by the administration of vitamin K. These findings provide insights into the prevention and management of drug-induced bleeding disorders. The established cell-based, high-throughput screening approach provides a powerful tool for identifying new vitamin K antagonists that function as anticoagulants.