A screening pipeline for antiparasitic agents targeting cryptosporidium inosine monophosphate dehydrogenase.
A screening pipeline for antiparasitic agents targeting cryptosporidium inosine monophosphate dehydrogenase.
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DOI：
10.1371/journal.pntd.0000794
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发表时间：
2010-08-10
影响因子：
3.8
通讯作者：
Striepen B
中科院分区：
文献类型：
--
作者：
Sharling L;Liu X;Gollapalli DR;Maurya SK;Hedstrom L;Striepen B
原生动物寄生虫隐孢子虫是发展中国家儿童重大疾病负担的原因。此外，隐孢子虫病可导致艾滋病患者的慢性和危及生命的肠炎，目前可用的药物在治疗这些严重疾病方面缺乏疗效。新的抗隐孢子虫疗法的发现和开发受到这种病原体的实验易处理性差的阻碍。虽然基因组测序工作已经确定了几个有趣的新目标，包括一个独特的肌苷一磷酸脱氢酶（IMPDH），追求这些目标和测试抑制剂一直令人沮丧的困难。在这里，我们开发了一系列工具来加速体内筛选C。小顶孢霉我们已经通过基因工程改造了相关的弓形虫作为C.细小病毒感染作为第一筛查。该检测提供了关键的靶标验证和目前在涉及C的检测中不可能实现的大信号窗口。小的为了进一步开发通过第一个过滤器的化合物，我们建立了基于荧光的宿主细胞增殖测定法，并建立了C。利用自动化高内容成像分析以提高通量的微小体生长测定。我们已经使用这些测定来评估C。parvum IMPDH抑制剂，并已鉴定出1，2，3-三唑醚的子集，其在T.弓形虫模型和提高的抗隐孢子虫活性。持续性腹泻是贫困儿童患病和死亡的主要原因，而这种疾病负担中越来越多的份额可归因于寄生虫隐孢子虫。没有疫苗可以预防隐孢子虫感染，治疗选择有限且不可靠。严重的是，对患有艾滋病的儿童或成人没有有效的治疗方法。隐孢子虫对药物发现提出了许多技术障碍;也许最重要的障碍是监测药物作用的困难。在这里，我们开发了一套方法来加速隐孢子虫病的药物发现过程。我们利用相关的寄生虫弓形虫的实验操作的机会，遗传工程隐孢子虫模型。这种新的寄生虫模型反映了隐孢子虫的代谢，为一个特别有前途的药物靶点提供了DNA和RNA的构建模块。药物有效性可以通过简单的荧光测量来测定许多候选人。使用这种测定作为初始过滤器，并使其他测定适应高通量格式，我们确定了几种新的化合物，表现出显着改善的抗隐孢子虫活性和优良的选择性。
The protozoan parasite Cryptosporidium parvum is responsible for significant disease burden among children in developing countries. In addition Cryptosporidiosis can result in chronic and life-threatening enteritis in AIDS patients, and the currently available drugs lack efficacy in treating these severe conditions. The discovery and development of novel anti-cryptosporidial therapeutics has been hampered by the poor experimental tractability of this pathogen. While the genome sequencing effort has identified several intriguing new targets including a unique inosine monophosphate dehydrogenase (IMPDH), pursuing these targets and testing inhibitors has been frustratingly difficult. Here we have developed a pipeline of tools to accelerate the in vivo screening of inhibitors of C. parvum IMPDH. We have genetically engineered the related parasite Toxoplasma gondii to serve as a model of C. parvum infection as the first screen. This assay provides crucial target validation and a large signal window that is currently not possible in assays involving C. parvum. To further develop compounds that pass this first filter, we established a fluorescence-based assay of host cell proliferation, and a C. parvum growth assay that utilizes automated high-content imaging analysis for enhanced throughput. We have used these assays to evaluate C. parvum IMPDH inhibitors emerging from our ongoing medicinal chemistry effort and have identified a subset of 1,2,3-triazole ethers that exhibit excellent in vivo selectivity in the T. gondii model and improved anti-cryptosporidial activity. Persistent diarrhea is a leading cause of illness and death among impoverished children, and a growing share of this disease burden can be attributed to the parasite Cryptosporidium. There are no vaccines to prevent Cryptosporidium infection, and the treatment options are limited and unreliable. Critically, no effective treatment exists for children or adults suffering from AIDS. Cryptosporidium presents many technical obstacles for drug discovery; perhaps the most important roadblock is the difficulty of monitoring drug action. Here we have developed a set of methods to accelerate the drug discovery process for cryptosporidiosis. We exploit the opportunities for experimental manipulation in the related parasite Toxoplasma to genetically engineer a Cryptosporidium model. This new model parasite mirrors the metabolism of Cryptosporidium for a particularly promising drug target that supplies the building blocks for DNA and RNA. Drug effectiveness can be assayed through simple fluorescence measurements for many candidates. Using this assay as an initial filter, and adapting other assays to a high throughput format, we identify several novel chemical compounds that exhibit markedly improved anti-cryptosporidial activity and excellent selectivity.
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感染期间淋巴结中中性粒细胞迁移的动态。
影响因子：
32.4
作者：
Chtanova, Tatyana;Schaeffer, Marie;Han, Seong-Ji;van Dooren, Giel G.;Nollmann, Marcelo;Herzmark, Paul;Chan, Shiao Wei;Satija, Harshita;Camfield, Kristin;Aaron, Holly;Striepen, Boris;Robey, Ellen A.
通讯作者：
Robey, Ellen A.
影响因子：
5.2
作者：
Gargala, G;Delaunay, A;Ballet, JJ
通讯作者：
Ballet, JJ
影响因子：
4.9
作者：
Downey, Autumn S.;Chong, Curtis R.;Sullivan, David J.
通讯作者：
Sullivan, David J.
影响因子：
4
作者：
De Koning, HP;Al-Salabi, MI;Wastling, JM
通讯作者：
Wastling, JM
影响因子：
4.9
作者：
Cai, XM;Woods, KM;Zhu, G
通讯作者：
Zhu, G