ANTIMALARIAL ACTION AND RESISTANCE

疟疾每年有2亿至3亿病例，200万至300万人死亡， 具有极其重要的医学意义 治疗和预防 由恶性疟原虫引起的疟疾由于对 氯喹（在南美洲、东南亚和非洲）， 缺乏确定的抗疟作用分子靶点。 这些研究的具体目的是：1）定义分子（S） 负责氯喹外排，从而对氯喹耐药性， P.恶性疟原虫，2）定义负责氯喹的分子 积累并因此对恶性疟原虫中氯喹的敏感性， 3）建立喹啉类化合物的构效关系 抗疟药 抗氯喹与敏感恶性疟原虫的杂交 将被用于识别负责 氯喹抗性 为了实现这一点，一个来自 抗性Dd 2亲本菌株将与来自 抗性后代 基于这些信息， 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。 分子（S）负责 氯喹的蓄积将通过其结合 氯喹使用：凝胶电泳和放射自显影，氯喹-- 琼脂糖柱层析，氯喹蓄积抑制 通过针对重构囊泡制剂的抗体，和 可光活化的交联试剂。 发展构效关系 具有特定结构的喹啉类似物 将测试修饰物抑制氯喹的能力 通过重构的囊泡制剂和用于 对氯喹敏感和抗性的抗疟原虫活性 菌株 这些研究的广泛的长期目标是提供一个合理的 发展抗疟药物的基础 多重抗性恶性疟原虫

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.