ANTIMALARIAL ACTION AND RESISTANCE

抗疟作用和耐药性

• 批准号: 3566974
• 负责人: Donald John Krogstad
• 金额: $ 21.87万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-09-01 至 1995-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3566974
• 关键词: Plasmodium falciparum; antimalarial agents; autoradiography; chemical binding; chemical structure function; chloroquine; drug design /synthesis /production; drug metabolism; gel electrophoresis; gene expression; human subject; laboratory mouse; laboratory rabbit; malaria vaccines; monoclonal antibody; multidrug resistance; nucleic acid hybridization; nucleic acid probes; peptide chemical synthesis; pharmacokinetics; pulsed field gel electrophoresis; restriction fragment length polymorphism; scintillation counter

With 200-300 million cases and 2-3 million deaths each year, malaria is of overwhelming medical importance. The treatment and prevention of malaria due to Plasmodium falciparum is complicated by resistance to chloroquine (in South America, Southeast Asia and Africa), and by the absence of defined molecular targets for antimalarial action. The specific aims of these studies are to: 1) define the molecule(s) responsible for chloroquine efflux and thus for chloroquine resistance in P. falciparum, 2) define the molecule(s) responsible for chloroquine accumulation and thus for susceptibility to chloroquine in P. falciparum, and 3) develop structure-activity relationships for the quinoline antimalarials. A cross between chloroquine-resistant and susceptible P. falciparum will be used to identify the gene(s) responsible for chloroquine-resistance. To accomplish this, a genomic DNA library from the resistant Dd2 parent strain will be hybridized to DNA from the resistant progeny. Based on this information peptides will be synthesized and used to produce antibodies to test for the gene product in resistant and susceptible strains. The molecule(s) responsible for chloroquine accumulation will be identified by its ability to bind chloroquine using: gel electrophoresis and autoradiography, chloroquine-- sepharose column chromatography, inhibition of chloroquine accumulation by antibodies to a reconstituted vesicle preparation, and photoactivatable cross-linking reagents. To develop structure-activity linking relationships, quinoline analogs with specific structural modifications will be tested for their ability to inhibit chloroquine accumulation by the reconstituted vesicle preparation and for antiplasmodial activity against chloroquine-susceptible and resistant strains. The broad long-term goal of these studies is to provide a rational basis for the development of antimalarials active against multiply-resistant P. falciparum.

疟疾每年有 200-3 亿病例和 2-300 万人死亡 具有压倒性的医学重要性。 治疗和预防 恶性疟原虫引起的疟疾因耐药性而变得复杂 氯喹（在南美洲、东南亚和非洲），以及 缺乏明确的抗疟作用分子靶点。 这些研究的具体目的是：1）定义分子 负责氯喹外流，从而导致氯喹耐药性 恶性疟原虫，2) 定义负责氯喹的分子 恶性疟原虫中的积累以及对氯喹的敏感性， 3) 建立喹啉的构效关系 抗疟药。 抗氯喹和易感恶性疟原虫之间的杂交 将用于识别负责的基因 氯喹耐药。 为了实现这一目标，基因组 DNA 文库来自 抗性 Dd2 亲本菌株将与来自 抗性后代。 根据这些信息，肽将被 合成并用于生产抗体以测试基因产物 在抗性和敏感菌株中。 负责的分子 氯喹的积累将通过其结合能力来确定 氯喹使用：凝胶电泳和放射自显影，氯喹-- 琼脂糖柱层析，抑制氯喹积累 通过针对重构的囊泡制剂的抗体，以及 光活化交联剂。 发展结构-活性 连接关系，具有特定结构的喹啉类似物 将测试修饰抑制氯喹的能力 重组囊泡制剂的积累和 针对氯喹敏感和耐药者的抗疟原虫活性 菌株。 这些研究的广泛长期目标是提供合理的 开发抗疟药的基础 多重耐药性恶性疟原虫。