Systematic Analysis of Clemastine, a Candidate Apicomplexan Parasite-Selective Tubulin-Targeting Agent.
Systematic Analysis of Clemastine, a Candidate Apicomplexan Parasite-Selective Tubulin-Targeting Agent.
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DOI：
10.3390/ijms23010068
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发表时间：
2021-12-22
影响因子：
5.6
通讯作者：
Morrissette NS
中科院分区：
文献类型：
--
作者：
Abbaali I;Truong DA;Day SD;Haro-Ramirez N;Morrissette NS
顶复门寄生虫，如刚地弓形虫，疟原虫属，巴贝虫属，和隐孢子虫属，导致显著的发病率和死亡率。由于毒性和抗药性寄生虫的出现，现有的治疗方法存在问题。由于原生动物微管蛋白可以被小分子选择性地破坏以抑制寄生虫生长，我们组装了一个体外测试级联，以充分描绘候选微管蛋白靶向药物对弓形虫和脊椎动物宿主细胞的影响。使用该分析，我们评估了氯马斯汀，这是一种抗组胺药，先前已显示通过竞争性结合CCT/TRiC微管蛋白伴侣抑制疟原虫生长作为概念验证。我们同时分析了阿司咪唑，一种独特的抗组胺剂，阻止血红素解毒疟原虫。这两种药物的EC 50值约为2 µM，在此浓度下未显示细胞毒性或脊椎动物微管破坏。用氯马斯汀或阿司咪唑治疗可缩短寄生虫膜下微管，但用乙胺嘧啶治疗后不缩短，表明这种效应不是抗寄生虫药物的一般反应。免疫印迹定量表明，氯马斯汀给药后，总α-微管蛋白浓度（0.02 pg/速殖子）未发生变化。总之，测试级联允许分析小分子对寄生虫和脊椎动物细胞活力和微管完整性的影响。
Apicomplexan parasites, such as Toxoplasma gondii, Plasmodium spp., Babesia spp., and Cryptosporidium spp., cause significant morbidity and mortality. Existing treatments are problematic due to toxicity and the emergence of drug-resistant parasites. Because protozoan tubulin can be selectively disrupted by small molecules to inhibit parasite growth, we assembled an in vitro testing cascade to fully delineate effects of candidate tubulin-targeting drugs on Toxoplasma gondii and vertebrate host cells. Using this analysis, we evaluated clemastine, an antihistamine that has been previously shown to inhibit Plasmodium growth by competitively binding to the CCT/TRiC tubulin chaperone as a proof-of-concept. We concurrently analyzed astemizole, a distinct antihistamine that blocks heme detoxification in Plasmodium. Both drugs have EC50 values of ~2 µM and do not demonstrate cytotoxicity or vertebrate microtubule disruption at this concentration. Parasite subpellicular microtubules are shortened by treatment with either clemastine or astemizole but not after treatment with pyrimethamine, indicating that this effect is not a general response to antiparasitic drugs. Immunoblot quantification indicates that the total α-tubulin concentration of 0.02 pg/tachyzoite does not change with clemastine treatment. In conclusion, the testing cascade allows profiling of small-molecule effects on both parasite and vertebrate cell viability and microtubule integrity.
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影响因子：
2.9
作者：
Khoury MK;Parker I;Aswad DW
通讯作者：
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56.9
作者：
CHAN, MMY;FONG, D
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作者：
Ma, Christopher;Tran, Johnson;Morrissette, Naomi
通讯作者：
Morrissette, Naomi
DOI：
10.1073/pnas.1913525117
发表时间：
2020-03-17
影响因子：
11.1
作者：
Lu, Kuan-Yi;Quan, Baiyi;Derbyshire, Emily R.
通讯作者：
Derbyshire, Emily R.