Bacterial-type hexokinase (HK) in the opportunistic parasite Cryptosporidium parv

机会性寄生虫隐孢子虫中的细菌型己糖激酶 (HK)

基本信息

批准号：
8898712
负责人：
GUAN ZHU
金额：
$ 17.78万
依托单位：
TEXAS A&M AGRILIFE RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-08-01 至 2017-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8898712
关键词：
Acquired Immunodeficiency Syndrome Animal Model Animals Bacterial Typing Biochemical Biological Assay Categories Cessation of life Chemicals Chlorine Cities Coccidia Communities Cryptosporidiosis Cryptosporidium Cryptosporidium parvum Data Development Diarrhea Diphosphates Disease Outbreaks Drug Targeting Drug resistance Entamoeba Enzymes Future Glycolysis Goals Growth Health Human Immunocompetent Immunocompromised Host In Vitro Individual Infection Kinetics Laboratories Lactate Dehydrogenase Lead Libraries Life Metabolic Metabolic Pathway Molecular Morbidity - disease rate Oocysts Parasites Pathway interactions Patients Pharmaceutical Preparations Process Protein Isoforms Proteins Reaction Resistance Side Stress Structure-Activity Relationship Testing Translational Research Trichomonas Trypanosoma United States United States National Institutes of Health Water Water Supply Wisconsin analog base biodefense drug development drug discovery enzyme activity foodborne foodborne pathogen gastrointestinal epithelium genome sequencing hexokinase high throughput screening inhibitor/antagonist mortality nitazoxanide novel therapeutics pathogen programs research study

项目摘要

DESCRIPTION: Cryptosporidium parvum is an AIDS-OI pathogen and Category B agent. It is a significant water- and food-borne protozoan parasite that can cause severe watery diarrhea in humans and animals. Currently, only a single drug (i.e., nitazoxanide [NTZ]) is approved for treating cryptosporidiosis in immunocompetent (but not immunocompromized) patients in the United States. However, NTZ is not 100% effective against cryptosporidiosis. Potential drug resistance may also be developed in Cryptosporidium, similar to the drug resistance problem in malarial parasites and coccidia. Therefore, there is an urgent need to develop new anti-Cryptosporidium drugs. Our long-term goal of this translational research is to characterize the molecular and biochemical features of the essential enzymes in the C. parvum glycolytic pathway as potential drug targets. Because C. parvum relies solely on glycolysis for its energy, we hypothesize that the bacterial-type CpHK responsible for the entry step of reaction in the glycolytic pathways could serve as rational drug targets in this parasite. To test the hypothesis, we will perform experiments to characterize the biochemical features of CpHK and identify CpHK inhibitors for future drug development via HTS of compound libraries, characterize the inhibitory kinetics of top hits, and test their efficacies against C. parvum growth in vitro. The major goal of this exploratory/developmental R21 project is to identify hits that inhibit both CpHK enzyme activity and the parasite growth in vitro, which would produce conclusive data for further studying the mechanism of inhibition, structure-activity relationship, and to synthesize new analogs of hits for lead optimization, as well as to assess the efficacies of hits and leads agains cryptosporidial infection in animal models.

描述：小隐孢子虫是一种 AIDS-OI 病原体，属于 B 类病原体。它是一种重要的水源和食源性原生动物寄生虫，可引起人类和动物严重的水样腹泻。目前，在美国，只有一种药物（即硝唑尼特 [NTZ]）被批准用于治疗免疫功能正常（但不是免疫功能低下）患者的隐孢子虫病。然而，NTZ 对隐孢子虫病并不是 100% 有效。隐孢子虫也可能产生潜在的耐药性，类似于疟疾寄生虫和球虫的耐药性问题。因此，迫切需要开发新的抗隐孢子虫药物。我们这项转化研究的长期目标是表征作为潜在药物靶点的 C. parvum 糖酵解途径中必需酶的分子和生化特征。由于 C. parvum 仅依赖糖酵解获取能量，因此我们假设负责糖酵解途径反应进入步骤的细菌型 CpHK 可以作为该寄生虫的合理药物靶标。为了检验这一假设，我们将进行实验来表征 CpHK 的生化特征，并通过化合物库的 HTS 鉴定用于未来药物开发的 CpHK 抑制剂，表征热门命中的抑制动力学，并测试它们在体外对 C. parvum 生长的功效。这个探索性/开发性 R21 项目的主要目标是确定抑制 CpHK 的命中酶活性和体外寄生虫生长，这将为进一步研究抑制机制、构效关系、合成新的命中类似物以优化先导化合物以及评估命中和先导化合物的功效提供结论性数据动物模型中的隐孢子虫感染。