Development of Flavivirus Antivirals Targeting Dengue Virus and WNV Protease

针对登革热病毒和西尼罗河病毒蛋白酶的黄病毒抗病毒药物的开发

• 批准号: 8845514
• 负责人: Sridhar G Prasad
• 金额: $ 30万
• 依托单位: PLEX PHARMACEUTICALS, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-15 至 2017-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8845514
• 关键词: Acute; Animal Model; Antiviral Agents; Arthralgia; Aspartic Endopeptidases; Binding; Bioavailable; Biochemical; Biological Assay; Biological Availability; Catechols; Categories; Cell Line; Cells; Centers for Disease Control and Prevention (U.S.); Cessation of life; Child; Country; Culicidae; Data; Dengue; Dengue Hemorrhagic Fever; Dengue Shock Syndrome; Dengue Virus; Development; Digit structure; Disease Outbreaks; Docking; Drug Kinetics; Encephalitis; Enzymes; Epidemic; Evaluation; Exanthema; Exhibits; Extravasation; FDA approved; Family; Fever; Flaviviridae; Flavivirus; Flavivirus Infections; Frequencies; Generations; Genome; Genomics; Goals; HIV-1; Hawaii; Health; Hemorrhage; Hepatitis C virus; High Pressure Liquid Chromatography; Housing; Human; In Vitro; Infection; Inhibitory Concentration 50; Japanese encephalitis virus; Lead; Libraries; Life; Liver; Mammalian Cell; Meningitis; Metabolism; Microsomes; Molecular Models; Morbidity - disease rate; N-terminal; National Institute of Allergy and Infectious Disease; Nonstructural Protein; PTPN11 gene; Paralysed; Patients; Peptide Hydrolases; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phase; Play; Polyproteins; Property; Protease Domain; Protein C; Proteins; Puerto Rico; RNA; RNA triphosphatase; Rattus; Recombinants; Replicon; Reporting; Research Proposals; Risk; Role; Safety; Series; Serine Protease; Serotyping; Severities; Shock; Solubility; Structural Protein; Structure; Structure-Activity Relationship; Supportive care; Symptoms; Therapeutic; Therapeutic Index; Toxic effect; Vero Cells; Viral; Virus Diseases; Virus-like particle; West Nile virus; Yellow fever virus; absorption; analog; base; burden of illness; channel blockers; chemotherapeutic agent; cofactor; cost; cytotoxicity; data modeling; design; drug development; enzyme substrate; helicase; in vitro activity; in vivo; inhibitor/antagonist; innovation; meetings; member; molecular modeling; mortality; novel; pathogen; scaffold; screening; small molecule; socioeconomics; three dimensional structure

DESCRIPTION (provided by applicant): Mosquito-borne members of the Flavivirus family include four serotypes of dengue virus (DENV1-4) and West Nile Virus (WNV) and are classified as re-emerging pathogens due to the frequency and severity of recent epidemics. There are an estimated 100 million DENV infections reported each year resulting in nearly ~25,000 deaths. More than 2.5 billion people are at risk of infection by DENV. Symptoms of dengue fever include fever, rash, and arthralgia. In more severe cases, viral infections can develop into life-threatening dengue hemorrhagic fever or dengue shock syndrome. Dengue shock syndrome occurs when leakage and/or bleeding is sufficient to induce shock, which often leads to death, especially in children, due to the lack of adequate supportive care. WNV infections can result in meningitis, encephalitis, or paralysis leading to ~10% mortality among hospitalized patients. Year 2012 witnessed a sudden surge in the number of WNV infections, comprising >5,000 cases in 48 states with ~228 deaths. Despite the high morbidity and mortality resulting from flavivirus infections in a subset of patients, there is currently no effective chemotherapeutic treatment for infections for any of the flaviviruses. The combined global socioeconomic impact of flavivirus pathogens merits an urgent need for new and effective antiviral therapeutics. The Flavivirus RNA genome encodes for three structural proteins (C, prM and E) and seven nonstructural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B and NS5). The NS3 protein is a multifunctional enzyme that contains protease, helicase and RNA triphosphatase activities. The N-terminal region of NS3 has a serine protease domain, which in the presence of the NS2B cofactor (NS2B, is an active serine protease. This protease (NS2B-NS3pro) plays an essential role in the cleavage of the viral precursor polyprotein and disruption of this function is lethal t viral replication. Therefore, blocking viral replication by inhibiting the quintessential viral protease could lead to the development of effective broad-spectrum antiviral therapeutics and is the focus of our research proposal. The innovation of our proposal is the discovery of two lead inhibitors, which have broad-spectrum (DENV1-4 and WNV) inhibitory activity and contain a catechol moiety, which is also a substructure present in 17 FDA approved drugs. In addition, the modeling data suggest the compounds bind to the enzyme substrate binding pocket and thus act as competitive inhibitors. Therefore, these properties of our leads support the feasibility for development into effective broad-spectrum antiviral therapeutics. The specific aims for Phase I of the proposal are: 1. Design and synthesize broad-acting inhibitors of DENV1-4 and WNV NS3pro; Milestone: Compounds with IC50d100nM against DENV1-4 and WNV protease will be advanced into cell-based assays. 2. Evaluate the therapeutic indices of optimized lead compounds using in vitro cell-based replicon, infectivity (EC50), and cytotoxicity (CC50) assays; Milestone: Identify 15-20 lead compounds with EC50d200nM and CC50e200uM representing the catechol and non-catechol series. 3. Conduct in vitro ADME-based lead optimization of compounds with broad-spectrum in vitro activity and acceptable therapeutic indices; Milestone: Identify 5-10 lead compounds exhibiting high bioavailability, weak inhibitor of CYPP450s, optimum stability and are not hERG channel blockers for evaluation in in vivo animal models for acute toxicity, efficacy and pharmacokinetics (Phase II application).

描述（由申请人提供）：Flavivirus家族的蚊子传播成员包括四种登革热病毒（DENV1-4）和西尼罗河病毒（WNV）的血清型，并被归类为由于最近流行病的频率和严重程度而被归类为重新出现的病原体。估计每年有1亿个DENV感染导致近25,000人死亡。 DENV有超过25亿人有感染的风险。登革热的症状包括热，皮疹和关节痛。在更严重的情况下，病毒感染可能会发展为威胁生命的登革热出血热或登革热休克综合征。当泄漏和/或出血足以引起冲击时，会发生登革热综合征，这通常导致死亡，尤其是在儿童中，由于缺乏足够的支持性护理。 WNV感染会导致脑膜炎，脑炎或麻痹，导致住院患者死亡约10％。 2012年的WNV感染数量突然激增，其中48个州的死亡人数约为5,000例。尽管一部分患者的黄病毒感染引起的发病率和死亡率很高，但目前尚无任何黄病毒感染的有效化学治疗方法。 Flavivirus病原体的全球社会经济综合影响值得迫切需要对新的和有效的抗病毒治疗。黄病毒RNA基因组编码三种结构蛋白（C，PRM和E）和7种非结构蛋白（NS1，NS2A，NS2B，NS2B，NS3，NS3，NS4A，NS4B和NS5）。 NS3蛋白是一种多功能酶，含有蛋白酶，解旋酶和RNA三磷酸酶活性。 NS3的N末端区域具有丝氨酸蛋白酶结构域，在NS2B辅因子（NS2B）存在下，该蛋白酶是一种活跃的丝氨酸蛋白酶。该蛋白酶（NS2B-NS3PRO）在病毒前的多蛋白质的裂解中起着至关重要的作用，从而阻止了这种函数的病毒性。典型的病毒蛋白酶可以导致有效的广谱抗病毒治疗学，这是我们的研究建议的重点。化合物结合酶底物结合袋，因此起到了竞争性抑制作用。 发展为有效的广谱抗病毒治疗剂。该提案的第一阶段的具体目的是：1。设计和合成DENV1-4和WNV NS3PRO的广泛作用抑制剂；里程碑：针对DENV1-4和WNV蛋白酶的IC50D100NM的化合物将被进展为基于细胞的测定。 2。使用基于体外细胞的复制子，感染性（EC50）和细胞毒性（CC50）测定法评估优化铅化合物的治疗指标；里程碑：用EC50D200NM和CC50E200UM识别15-20个铅化合物，代表Catechol和非Catechol系列。 3。在体外基于ADD的铅铅优化，对具有广谱的体外活性和可接受的治疗指标进行了铅的优化；里程碑：鉴定5-10种铅化合物，表现出高生物利用度，CYPP450的抑制剂较弱，最佳稳定性，并且不是用于在体内动物模型中评估急性毒性，功效和药物动力学（II期应用）的HERG通道阻滞剂。