Functional Genomic Approach to Macrofilaricide Discovery

描述（由申请人提供）：寄生丝虫病引起淋巴丝虫病和河盲症（盘尾丝虫病），在至少80个国家和地区感染超过1.4亿人，其中高达10亿人处于危险之中。淋巴丝虫病（由乌氏丝虫病和布鲁氏丝虫病引起）在受影响地区造成巨大的健康和经济负担，是造成长期残疾的第二大原因。2000年，包括世界卫生组织在内的国际组织发起了一项倡议，通过大规模给药来消除淋巴丝虫病。虽然该项目中可用的化合物（伊维菌素、阿苯达唑、二乙基卡马嗪）对杀死幼虫（微丝虫）有效，但寄生虫（大丝虫）的成虫库存活并可在体内停留10年。此外，大规模治疗的极端选择性压力导致最近出现了寄生虫耐药性的迹象。用伊维菌素减少盘尾丝虫病的运动也面临着类似的挑战。该项目的长期目标是开发具有“大丝杀虫”活性的新化合物，以补充目前的治疗方案。在第一阶段的研究中，利用模型线虫秀丽隐杆线虫和马来布鲁氏丝虫病的生物信息学过滤器和功能基因组（RNAi）数据，成功地识别并优先考虑了一小组预测对大丝虫病生存至关重要的分子靶点。此外，作为II期研究的前奏，我们对先前在Divergence上被证明具有杀线虫活性的化合物进行了体外对Brugia spj的杀线虫活性测试，其中一些化合物显示出有希望的杀线虫效果。在当前的研究中，我们将通过高通量化学文库筛选和靶向化学信息学方法的结合，表达丝线虫生物合成途径中两种特性良好且必需的酶，并鉴定抑制剂。将对最佳候选物进行体外和成虫马来芽孢杆菌的鉴定，并测定其在哺乳动物体内的细胞毒性和微粒体稳定性。最后，最有希望的化合物将进入淋巴丝虫病的啮齿动物模型试验。这项工作的预期结果是鉴定出在动物模型中表现出大丝虫杀灭活性的铅分子。在与已表明致力于全球卫生公平的机构（如盖茨基金会或世卫组织）的合作下，三期工作将侧重于启动人体临床试验——最终目标是消除丝虫病对人类健康的负担。（与公共卫生有关）寄生线虫（蛔虫）目前感染全球超过10亿人，造成严重发病率和相当大的经济损失。该项目旨在确定迫切需要的药物，以对抗构成最严重的人类热带聚集性疾病的丝状线虫。丝虫病引起一些已知的更有害的线虫传播疾病（象皮病和河盲症）。

DESCRIPTION (provided by applicant): Parasitic filarial nematodes causing lymphatic filariasis and river blindness (onchocerciasis), infect over 140 million people with up to one billion at risk in at least 80 countries and territories. Lymphatic filariasis (caused by Wuchereria and Brugia species) creates an enormous health and economic burden within affected areas and is ranked as the second leading cause of long-term disability. In 2000, international groups including the World Health Organization launched an initiative to eliminate lymphatic filariasis by conducting mass drug administrations. Although the compounds available for this program (ivermectin, albendazole, diethylcarbamazine) are effective at killing larvae (microfilariae), the adult reservoir of parasites (macrofilariae) survive and can remain in the body for a decade. In addition, the extreme selective pressure of mass treatment has lead to recent signs of parasite drug resistance. Similar challenges face the campaigns to reduce onchocerciasis with ivermectin. The long-term objective of this project is to develop new compounds with 'macrofilaricidal' activity that are desperately needed to complement the current regimens. During Phase I efforts, bioinformatic filters and functional genomic (RNAi) data from the model nematode Caenorhabitis elegans and the filarial parasite Brugia malayi were utilized to successfully identify and prioritize a small set of molecular targets predicted to be critical for macrofilarial survival. In addition, as a prelude to Phase II research, compounds previously demonstrated to possess nematicidal activity at Divergence were tested against Brugia spp. in vitro, with some compounds showing promising macrofilaricidal effects. In the current research, two well-characterized and essential enzymes from a filarial nematode biosynthetic pathway will be expressed and inhibitors identified through a combination of high-throughput chemical library screening and targeted cheminformatic approaches. The best hits will be evaluated against adult B. malayi and in vitro and assayed for mammalian cytotoxicity and microsomal stability. Finally, the most promising compounds will progress into rodent model trials of lymphatic filariasis. The expected outcome of this work is the identification of lead molecules that demonstrate macrofilaricidal activity within animal models. In collaboration with agencies that have demonstrated a commitment to global health equity (such as the Gates Foundation or WHO), Phase III efforts will focus on the initiation of clinical trials in humans--with the ultimate goal of eliminating the human health burden of filarial diseases. (Relevance to public health) Parasitic nematodes (roundworms) currently infect over 1 billion people globally, causing severe morbidity and considerable economic losses. This project is structured to identify desperately needed drugs to combat filarial nematodes, which constitute the most significant of the human tropical cluster diseases. Filarial parasites cause some of the more pernicious nematode-borne diseases known (elephantiasis and river blindness).