Identification of selective inhibitors of phosphofructokinase as lead compounds a

描述（由申请人提供）：本项目的目的是发现和优化锥虫（布氏锥虫，T. cruzi和利什曼原虫物种）。这些病原体在热带和亚热带国家主要是非洲、中美洲和南美洲以及亚洲引起严重的、往往致命的人类疾病，如昏睡病、恰加斯病和黑热病，在这些国家，数百万人生活在这些疾病流行的地区。不幸的是，目前用于治疗它们的药物不能令人满意，因为它们对某些形式的疾病是有毒的和无效的，并且耐药性变得越来越普遍。 糖酵解是T.因此是一个有前途的药物靶点。 因此，糖酵解酶抑制剂如PFK可作为开发新药的先导化合物。本研究的主要目的是：（1）利用锥虫PFK蛋白的独特结构（其结构信息已被证实，并已被RNAi证实为药物靶点），寻找锥虫PFK蛋白的选择性抑制剂。通过高通量筛选包含超过30万个小分子的分子库小分子库（MLSMR），（2）为了在一组二次命中验证测定中确认这些化合物的效力，并在动力学和选择性测定中测试它们的特异性，并进一步提高通过基于结构的方法获得的最有希望的分子的效力，类似物合成和药物化学原理;（3）通过酶分析、蛋白质晶体学和生物物理学测量确定最有希望的分子的作用模式;和（4）测试化合物显示出最高效力，即它们抑制代表寄生虫致病阶段的培养锥虫细胞生长的能力，以及对培养的人类细胞没有毒性。 公共卫生相关性：撒哈拉以南非洲的昏睡病、中美洲和南美洲的恰加斯病以及世界热带和亚热带地区的黑热病和相关疾病给已经陷入贫困、营养不良和疾病悲惨循环的人口造成了严重的公共卫生和经济负担。 全世界有数百万人感染了这些可能致命的疾病，数亿人处于危险之中。现有的治疗方法在过去40年中发展很少，并且存在毒性，效率低下和耐药性;该项目的目标是开发适合进入临床前试验的先导药物。

DESCRIPTION (provided by applicant): The goal of this project is to discover and optimize selective inhibitors of phosphofructokinase (PFK) of trypanosomatid parasites (Trypanosoma brucei, T. cruzi and Leishmania species). These pathogens cause serious, often fatal diseases of humans such as sleeping sickness, Chagas' disease and kala-azar in tropical and subtropical countries primarily in Africa, Central and South America, and Asia where many millions live in areas where the diseases are endemic. Tragically, current drugs for their treatment are unsatisfactory because they are toxic and ineffective against some forms of the diseases, and resistance is becoming increasingly common. Glycolysis is essential in the infective stage of T. brucei and therefore a promising drug target. Inhibitors of glycolytic enzymes such as PFK may thus serve as lead compounds for the development of new drugs. The proposed research has as specific aims: (1) To exploit unique features of trypanosomatid PFK (for which detailed structural information is already available and which has been validated as a drug target by RNAi) for the discovery of selective inhibitors of PFKs from T. brucei (TbPFK) through high-throughput screening of the Molecular Library Small Molecule Repository (MLSMR) containing more than 300,000 small molecules; (2) To confirm the potency of these compounds in a panel of secondary hit validation assays and to test their specificity in kinetic and selectivity assays, and to further improve the potency of the most promising molecules thus obtained by structure-based methods, analogue synthesis and medicinal chemical principles; (3) To determine the mode of action of the most promising molecules by enzyme assays, protein crystallography and biophysical measurements; and (4) To test compounds displaying the highest potency for their ability to inhibit growth of cultured trypanosomatid cells representing pathogenic stages of the parasites, as well as for lack of toxicity toward cultured human cells. PUBLIC HEALTH RELEVANCE: Sleeping sickness in sub-Saharan Africa, Chagas' disease in Central and South America, and kala-azar and related diseases in tropical and subtropical regions of the world cause severe public health and economic burdens on populations that are already caught in a tragic cycle of poverty, poor nutrition and disease. Millions of people worldwide are infected by these potentially fatal diseases and hundreds of millions are at risk. Existing treatments have developed little in the past 40 years, and suffer from toxicity, inefficiency and resistance; the goal of this project is to develop lead drugs that will be suitable for entry into pre-clinical trials.