Development of Novel MetAP2 inhibitors as potential therapeutics for Microsporidiosis

开发新型 MetAP2 抑制剂作为微孢子虫病的潜在疗法

• 批准号: 10408440
• 负责人: Bhaskar Chandra Das
• 金额: $ 39万
• 依托单位: LONG ISLAND UNIVERSITY BROOKLYN CAMPUS
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10408440
• 关键词: Acquired Immunodeficiency Syndrome; Antiparasitic Agents; Binding; Bioinformatics; Biological Assay; Biological Availability; Bone Marrow; Categories; Category B pathogen; Cells; Characteristics; Clinical; Crystallization; Data; Development; Diarrhea; Disease; Dose; Drug Design; Drug Targeting; Encephalitozoon cuniculi; Endothelial Cells; Enterocytozoon bieneusi; Enzyme Inhibition; Enzymes; Eukaryota; Food; Gastrointestinal tract structure; Genes; Goals; Grant; Growth; Human; Immune; Immunocompetent; In Vitro; Industry; Infection; Invertebrates; Lead; Leishmania donovani; Libraries; Mammals; Mass Spectrum Analysis; Microsporidia; Microsporidiosis; Modeling; Morbidity - disease rate; Nosema corneum; Opportunistic Infections; Organism; Parasitology; Pathogenicity; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacology; Plasmodium falciparum; Process; Property; Protozoa; Publications; Recombinants; Research; Resistance; Respiratory Muscles; Saccharomyces cerevisiae; Structure; Structure-Activity Relationship; Testing; Therapeutic; Therapeutic Agents; Thrombocytopenia; Toxic effect; United States National Institutes of Health; Vertebrates; Water; Yeasts; Zoonoses; analog; base; burden of illness; design; drug development; drug structure; emerging pathogen; fumagillin; high throughput screening; improved; in silico; in vitro testing; in vivo; in vivo Model; inhibitor/antagonist; interest; methionyl aminopeptidase; mortality; mouse model; nervous system infection; novel; novel drug class; novel therapeutics; opportunistic pathogen; pathogen; pharmacokinetics and pharmacodynamics; preclinical development; reproductive; scaffold; screening; structural biology; therapeutic target; therapy design; tool

Abstract This application represents an approach to rational drug design for the treatment of microsporidiosis using structure activity relationships (SAR) for agents that inhibit Methionine Aminopeptidase type 2 (MetAP2). Microsporidiosis is an emerging zoonotic infection that is an opportunistic pathogen in the setting of AIDS, but is also seen in other immune compromised hosts as well as immune competent hosts. Current therapies are suboptimal. The hypothesis underlying this type of grant is that differences in the structure of the drug target between host and pathogen will permit the design of selective therapeutic agents with decreased host toxicity. The initial choice in drug design is the selection of the target among the dozens of potential targets. MetAP2 is an extremely logical therapeutic target for these pathogens. Microsporidia lack Methionine Aminopeptidase type 1 (MetAP1) making MetAP2 an essential enzyme. Among eukaryotes this makes them highly susceptible to MetAP2 inhibitors and limits the toxicity of these compounds in their hosts as humans have both MetAP1 and MetAP2. Use of Fumagillin and its derivatives, which are non-competitive inhibitors that covalently bind to and inhibit MetAP2 (but not MetAP1), has confirmed that inhibition of MetAP2 is an effective in vitro and in vivo therapeutic target for many species of microsporidia. In fact, Fumagillin has been demonstrated to have efficacy in humans infected with Enterocytozoon bieneusi; however, its use has been limited by bone marrow toxicity. Our research group has cloned, expressed and determined the crystal structure of the MetAP2 of the microsporidian Encephalitozoon cuniculi (i.e. EcMetAP2) as well as developed yeast dependent on EcMetAP2 for growth. We have identified and cloned Ent.bieneusi MetAP2 (EbMetAP2). Exploiting differences in the structure of MetAP2 between host and pathogen should permit the design of selective therapeutic competitive inhibitors of MetAP2 with decreased host toxicity. These new inhibitors will be tested in vitro and in vivo for efficacy and in an iterative process we will use this information to refine our models and improve inhibitor design. Using this Limited Rational Design (LRD) approach we will generate new libraries based on our lead compounds that will then be modified to generate new libraries to improve their selectivity and pharmacologic properties. We have already identified two lead compounds, BL6 and D63, from our initial LRD/SAR studies that have increased selectivity for microsporidian MetAP2 and efficacy in our in vitro and in vivo models of microsporidiosis. Since MetAP2 is important in other protozoa our lead compounds should also define new classes of drugs that could have broad anti-parasitic activity and prove useful in the treatment of other infections. Our assembled research group containing experts in medicinal chemistry, parasitology, bioinformatics, mass spectrometry and structural biology as well as industry consultants has the necessary complementary expertise to develop and test these compounds.

摘要 本申请代表了一种合理设计药物治疗微孢子虫病的方法， 抑制甲氨酰氨肽酶2型（MetAP 2）药物的构效关系（SAR）。 微孢子虫病是一种新出现的人畜共患传染病，是艾滋病背景下的机会致病菌， 也见于其他免疫受损宿主以及免疫活性宿主。目前的治疗方法是 次优这种资助背后的假设是，药物靶点结构的差异 宿主和病原体之间的相互作用将允许设计具有降低的宿主毒性的选择性治疗剂。 药物设计的最初选择是从几十个潜在的靶点中选择靶点。MetAP 2是 是这些病原体的一个非常合理的治疗目标。微孢子虫缺乏甲氨酰氨肽酶 1型（MetAP 1）使MetAP 2成为必需酶。在真核生物中， 并限制这些化合物在其宿主中的毒性，因为人类既具有MetAP 1 MetAP2。夫马洁林及其衍生物的用途，所述夫马洁林及其衍生物是共价结合 并且抑制MetAP 2（但不抑制MetAP 1），已经证实抑制MetAP 2在体外和体内是有效的 是许多微孢子虫种类的治疗靶点。事实上，烟曲霉素已被证明具有 在感染比氏肠细胞虫的人类中有效;然而，其使用受到骨髓的限制， 毒性我们的研究小组已经克隆、表达并确定了MetAP 2的晶体结构。 微孢子虫兔脑孢子虫（即EcMetAP 2）以及依赖于EcMetAP 2的发育酵母 促进增长。我们已经鉴定并克隆了Ent.bieneusi MetAP 2（EbMetAP 2）。利用不同的 宿主和病原体之间的MetAP 2结构应该允许设计选择性治疗竞争性药物。 MetAP 2抑制剂，具有降低的宿主毒性。这些新的抑制剂将在体外和体内进行测试， 在迭代过程中，我们将使用这些信息来完善我们的模型并改进抑制剂 设计使用这种有限理性设计（LRD）方法，我们将根据我们的线索生成新的库 这些化合物然后将被修饰以产生新的文库，以提高它们的选择性和药理学活性。 特性.我们已经从我们最初的LRD/SAR研究中确定了两种先导化合物BL 6和D 63 在我们的体外和体内模型中， 微孢子虫病由于MetAP 2在其他原生动物中很重要，我们的先导化合物也应该定义新的 这类药物可能具有广泛的抗寄生虫活性，并证明可用于治疗其他 感染.我们的研究小组包括药物化学，寄生虫学， 生物信息学、质谱和结构生物学以及行业顾问有必要 开发和测试这些化合物的补充专业知识。