Small molecule protein-glycan inhib. as malaria transmission-blocking therapuetic

小分子蛋白聚糖抑制。

基本信息

批准号：
8237055
负责人：
Rhoel David Ramos Dinglasan
金额：
$ 40.18万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-04-22 至 2014-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8237055
关键词：
Acute Adhesions Affinity Affinity Chromatography Anabolism Animals Anopheles Genus Anopheles gambiae Antibodies Antibody Specificity Antigens Antimalarials Applications Grants Back Bacterial Adhesins Basic Science Binding Binding Sites Biological Assay Biological Availability Blocking Antibodies Candidate Disease Gene Carbohydrates Cell surface Cells Chemicals Chondroitin Chondroitin Sulfate Proteoglycan Chondroitin Sulfates Chronic Complement Complex Computer Simulation Concanavalin A Crystallization Crystallography Culicidae Data Development Disaccharides Disease Progression Dose Drug Compounding Drug Design Drug Kinetics Drug resistance Enzyme-Linked Immunosorbent Assay Enzymes Exhibits Face Gene Targeting Generations Genes Glycoconjugates Glycosaminoglycans Goals Hepatocyte Homology Modeling Housing Human Immune Sera Immunofluorescence Immunologic In Vitro Incubated Individual Invaded Knock-out Knowledge Lead Lectin Length Libraries Lichen - organism Life Cycle Stages Ligands Liquid Chromatography Liver Malaria Mammals Mass Spectrum Analysis Mediating Mediator of activation protein Membrane Methods Midgut Modeling Modification Molecular Molecular Models Molecular Target Mus Organelles Oryctolagus cuniculus Outsourced Services Parasites Pathway interactions Peptides Pharmaceutical Preparations Pharmacodynamics Plasmodium Plasmodium berghei Plasmodium falciparum Play Polymers Polysaccharides Population Post-Translational Protein Processing Preparation Process Production Progress Reports Property Protein Array Proteins Proteoglycan Proteomics RNA Interference Recombinant Proteins Recombinants Regimen Research Project Grants Roentgen Rays Role Route Safety Serum Sporozoites Staging Staining method Stains Streptavidin Structure Surface Surface Plasmon Resonance System Testing Therapeutic Tissues Toxic effect Transferase Translational Research Vaccines Variant Von Willebrand Factor A Domain Work absorption analog base cellular microvillus chondroitin sulfate glycosaminoglycan circumsporozoite design feeding glycosylation homologous recombination improved in vivo inhibitor/antagonist insight knockout gene man microbial mimetics molecular dynamics molecular modeling novel parasite invasion pathogen polypeptide pre-clinical preclinical study prevent protein expression research study response scaffold small molecule tandem mass spectrometry transmission process usnic acid vaccine candidate vector mosquito xylosyltransferase

项目摘要

DESCRIPTION (provided by applicant): Malaria transmission entails development of the Plasmodium parasite in the mosquito. We have identified a critical interaction between an unknown ookinete lectin-like protein and a chondroitin sulfate glycosaminoglycan ligand on the mosquito midgut lumenal surface. We hypothesize that by disrupting this interaction through the use of small molecule inhibitors we can prevent parasite establishment in the mosquito and, subsequently, completely abrogate malaria transmission. This is a translational research grant proposal with the goal of taking our basic research understanding of Plasmodium-mosquito host interactions toward the development of novel highly potent malaria transmission-blocking therapeutics. Our first aim, the Complete molecular characterization of Plasmodium ookinete protein-midgut glycosaminoglycan interactions involves (1) identifying novel lectin-like ookinete molecules by glycan-affinity chromatography and mass spectrometry, (2) characterizing their functional role in vivo through the production of gene knockout parasites, (3) assessing their binding affinity for mosquito chondroitin glycosaminoglycans by protein array-surface plasmon resonance, and (3) gaining insight into the structure-function of the molecule(s) in complex with chondroitin glycosaminoglycan fragments and structural analogues by molecular modeling and x-ray crystallography. The second aim of the project, the Development of lead Plasmodium transmission-blocking glycan-mimetic compounds and assessment of their transmission-blocking potential involves identification of novel derivatives and analogues of our lead transmission-blocking compound, VS1 (a non-peptidyl polyvinylsulfonated polymer), which is a structural mimic of midgut chondroitin glycosaminoglycans and inhibits >95% of parasite development in the mosquito. To develop more potent structural analogs, we propose a four-tiered approach: (1) isolation of varying chain-lengths of the VS1 polymer, (2) derivitization of VS1 to enhance inhibitory activity and bioavailability, (3) derivitization of (+)-usnic acid, a polyphenolic compound from lichens, and (4) assessment of the utility of peptide mimotopes of mosquito chondroitin sulfate glycosaminoglycans as transmission-blocking vaccine targets. To help progress toward preclinical studies, the top candidate compounds from each approach will be analyzed for their pharmacokinetic and pharmacodynamic properties in animal and human serum models. Mosquitoes transmit the Plasmodium parasite from infected human hosts to uninfected individuals. By blocking a critical protein-glycan (carbohydrate) interaction between the parasite and the mosquito vector through the use of small inhibitory compounds (drugs) we prevent parasite establishment in the mosquito and effectively disrupt the transmission cycle of the malaria parasite. This strategy is especially important given that many current antimalarials and vaccine candidates still permit transmission of the parasite through the mosquito, moreover, our lead compound has added potential utility as a multi-stage antimalarial therapeutic since it also prevents sporozoite establishment in liver cells, the first step towards the progression of disease in humans.

描述（由申请人提供）：疟疾传播需要在蚊子中发展疟原虫。我们已经确定了未知的Ookinete凝集素样蛋白与硫酸软骨蛋白糖胺聚糖配体在蚊子中肠腔表面上的关键相互作用。我们假设通过使用小分子抑制剂来破坏这种相互作用，我们可以防止蚊子中的寄生虫建立，然后完全消除疟疾的传播。这是一项翻译研究赠款建议，其目的是将我们对疟原虫 - 摩斯基托宿主相互作用的基础研究理解发展，以开发新型高度有效的疟疾传播阻断疗法。我们的第一个目的是，元素蛋白质蛋白 - 米形糖糖胺聚糖相互作用的完整分子表征涉及（1）鉴定新型的凝集素样植物分子，通过聚糖亲和力色谱和质谱法，（2）评估其在Vivo中的作用，以评估其在基因上的作用（3）（3）（2）蛋白阵列表面等离子体共振的软骨素糖胺聚糖，以及（3）通过分子模型和X射线晶体结晶的结构与软骨素糖胺聚糖片段和结构类似物的复合物中分子的结构功能（s）的结构功能。 The second aim of the project, the Development of lead Plasmodium transmission-blocking glycan-mimetic compounds and assessment of their transmission-blocking potential involves identification of novel derivatives and analogues of our lead transmission-blocking compound, VS1 (a non-peptidyl polyvinylsulfonated polymer), which is a structural mimic of midgut chondroitin glycosaminoglycans and inhibits在蚊子中> 95％的寄生虫发育。为了开发更有效的结构类似物，我们提出了一种四层方法：（1）隔离VS1聚合物的不同链长度的分离，（2）VS1的衍生化以增强抑制活性和生物利用度，（3）（3）从LICHENOCID和（usnic酸）评估（4）的（4）含量（4）的（4）的（4）。蚊子软骨素硫酸盐糖胺聚糖作为透射阻塞疫苗靶标。为了帮助临床前研究的进展，将分析每种方法中最高的候选化合物，以便在动物和人血清模型中的药代动力学和药效学特性。蚊子将寄生虫从感染的人类宿主传播到未感染的个体。通过通过使用小型抑制性化合物（药物）来阻止寄生虫和蚊子载体之间的关键蛋白质 - 甘露糖（碳水化合物）相互作用，我们可以防止蚊子中的寄生虫建立，并有效破坏疟原虫寄生虫的传播周期。考虑到许多当前的抗疟疾和疫苗候选物仍然允许通过蚊子传播该策略，因此，我们的铅化合物作为一种多阶段的抗疟药，因为它也可以阻止肝细胞中的孢子虫建立，这是肝脏疾病的第一步，这是人类疾病疾病的第一步。