CYP51 is an essential drug target for the treatment of primary amoebic meningoencephalitis (PAM).
CYP51 is an essential drug target for the treatment of primary amoebic meningoencephalitis (PAM).
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DOI：
10.1371/journal.pntd.0006104
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发表时间：
2017-12-01
影响因子：
3.8
通讯作者：
Podust, Larissa M
中科院分区：
文献类型：
--
作者：
Debnath, Anjan;Calvet, Claudia M;Podust, Larissa M
原发性阿米巴脑膜脑炎（PAM）是由福氏耐格里阿米巴引起的，福氏耐格里阿米巴是一种自由生活的阿米巴，偶尔会感染人类。虽然被认为是“罕见的”（但可能被低估），但高死亡率和缺乏既定的治疗成功率使PAM成为一种特别具有破坏性的感染。在制药行业缺乏投资开发抗PAM药物的经济诱因的情况下，抗PAM药物的发现主要依赖于药物“再开发”-一种应用已知药物治疗罕见或被忽视疾病的成本有效策略。与真菌相似，N. Fowleri对麦角甾醇（一种血浆和细胞膜的结构单元）具有基本需求。通过小分子抑制剂破坏甾醇生物合成是针对具有医学和农业重要性的真菌病原体的经验证的干预策略。了N. fowleri基因组编码甾醇14-脱甲基酶（CYP 51）靶标，与真菌直向同源物具有约35%的序列同一性。靶点的相似性提高了重新利用抗真菌药物和优化其用于治疗PAM的可能性。在这项工作中，我们（i）系统地评估了抗真菌唑类药物，称为康唑类，对甾醇生物合成和培养的N。fowleri滋养体，（ii）通过质谱分析N. fowleri脂质，和（iii）分析了重组CYP 51目标和conazoles之间的相互作用，通过紫外可见光谱和X-射线晶体学。总的来说，这些研究中获得的基于靶标和基于寄生虫的数据验证了CYP 51作为N. fowleri和康唑类药物作为PAM动物模型中评估的候选药物。
Primary Amoebic Meningoencephalitis (PAM) is caused by Naegleria fowleri, a free-living amoeba that occasionally infects humans. While considered "rare" (but likely underreported) the high mortality rate and lack of established success in treatment makes PAM a particularly devastating infection. In the absence of economic inducements to invest in development of anti-PAM drugs by the pharmaceutical industry, anti-PAM drug discovery largely relies on drug 'repurposing'-a cost effective strategy to apply known drugs for treatment of rare or neglected diseases. Similar to fungi, N. fowleri has an essential requirement for ergosterol, a building block of plasma and cell membranes. Disruption of sterol biosynthesis by small-molecule inhibitors is a validated interventional strategy against fungal pathogens of medical and agricultural importance. The N. fowleri genome encodes the sterol 14-demethylase (CYP51) target sharing ~35% sequence identity to fungal orthologues. The similarity of targets raises the possibility of repurposing anti-mycotic drugs and optimization of their usage for the treatment of PAM. In this work, we (i) systematically assessed the impact of anti-fungal azole drugs, known as conazoles, on sterol biosynthesis and viability of cultured N. fowleri trophozotes, (ii) identified the endogenous CYP51 substrate by mass spectrometry analysis of N. fowleri lipids, and (iii) analyzed the interactions between the recombinant CYP51 target and conazoles by UV-vis spectroscopy and x-ray crystallography. Collectively, the target-based and parasite-based data obtained in these studies validated CYP51 as a potentially 'druggable' target in N. fowleri, and conazole drugs as the candidates for assessment in the animal model of PAM.