Developing Combination Therapies against Pneumo- and Paramyxoviruses Causing Severe Respiratory Infection

开发针对引起严重呼吸道感染的肺炎病毒和副粘病毒的联合疗法

• 批准号: 10421071
• 负责人: Richard K. Plemper
• 金额: $ 53.93万
• 依托单位: GEORGIA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10421071
• 关键词: 1 year old; Achievement; Address; Antibodies; Award; Biochemical; Biological Assay; Cell Culture Techniques; Cell Line; Cells; Cellular Assay; Cessation of life; Child; Clinical; Combined Modality Therapy; Communicable Diseases; Complex; Cotton Rats; Development; Disease; Disease Management; Disease model; Drug Combinations; Drug Kinetics; Elderly; Engineering; Ensure; Failure; Family; Fluorescence Polarization; Foundations; Frequencies; Genetic; Genome; Henipavirus; Hospitalization; Human; Immunocompromised Host; In Vitro; Individual; Infant; Infection; Interferometry; Interphase; Lead; Libraries; Link; Lower Respiratory Tract Infection; Measles virus; Modeling; Mumps virus; Mus; Paramyxovirus; Patients; Pharmaceutical Preparations; Pilot Projects; Pneumovirus; Polymerase; Positioning Attribute; Property; Prophylactic treatment; Proteins; Public Health; Quantitative Structure-Activity Relationship; RNA Polymerase Inhibitor; RNA Viruses; RNA-Directed RNA Polymerase; Recombinants; Reporter; Resistance; Resistance profile; Respiratory Tract Infections; Respiratory syncytial virus; Ribonucleoproteins; Risk; Safety; Site; Structure-Activity Relationship; System; Technology; Testing; Therapeutic; Therapeutic Intervention; Time; Toxic effect; United States; Vaccines; Validation; Viral; Virus; Virus Diseases; Zoonoses; airway epithelium; analog; animal efficacy; attenuation; base; cost effective; counterscreen; deep sequencing; drug development; high risk; high throughput screening; human morbidity; human mortality; human pathogen; immunoprophylaxis; improved; in silico; in vivo; infant infection; inhibitor; innovation; lead candidate; member; next generation; novel; nucleoside analog; nucleoside inhibitor; pathogen; patient population; pediatric patients; pharmacophore; programs; prophylactic; resistance mechanism; resistance mutation; respiratory virus; scaffold; screening; small molecule therapeutics; sound; success; synergism; therapeutic target; therapeutically effective; viral RNA

It is the overarching hypothesis of this project that long-term therapeutic success against human parainfluenzavirus type 3 (HPIV3) and respiratory syncytial virus (RSV) will require a combination therapy approach with a pair of pathogen-directed inhibitors with distinct mechanistic profile. This notion is driven by the strict safety profile requested by a mostly pediatric patient population and the threat that resistance mutations against individual therapeutics may become fixed rapidly in circulating virus strains. Members of the closely related paramyxo- and pneumovirus families, HPIV3 and RSV are responsible for the majority of severe lower respiratory infection (LRI) and death from viral disease among infants in the United States, and recognized as a potential threat to the immunocompromised and the elderly. Infection by both pathogens initiates in the upper respiratory epithelium, followed by gradual progression to the small airways in patients advancing to severe disease, opening a window for therapeutic intervention. No vaccine protection or effective therapeutic is currently available against either HPIV3 or RSV, and antibody immunoprophylaxis against RSV is restricted to a subset of high-risk patients. This project will address this unmet clinical need by developing applicable, cost-effective therapeutics targeting the viral RNA-dependent RNA polymerase (RdRp) complexes. Building on an established antiviral program, we have recently identified an efficacious nucleoside analog inhibitor with potent activity against both RSV and HPIV3. Serving as reference, this compound will inform the co-development of allosteric RdRp blockers that are rigorously vetted from early stage development for their potential for combination therapy with competitive polymerase inhibitors. In pilot studies, we have engineered a recombinant HPIV3 reporter virus and identified in a high-throughput screening (HTS) campaign using this strain two novel, viable HPIV3 polymerase inhibitor scaffolds with sub-micromolar starting potency. Against RSV RdRp, we have synthetically redesigned an efficacious allosteric measles virus polymerase blocker and identified in the resulting pharmacophore-informed library a potent new point-of-entry with anti-RSV RdRp activity. Recognizing the risk of early stage failure in drug development, we have in parallel identified the protein-protein interface between the RSV polymerase and encapsidated genome as a promising yet underexplored druggable site. To further diversify the portfolio also of allosteric anti-RSV candidates, this site will be interrogated with an innovative biochemical HTS assay and orthogonal counterscreens (aim 1). The existing anti-RSV and anti-HPIV3 leads and newly emerging candidates will be mechanistically characterized using next-generation cell based and in vitro polymerase assays, and subjected to resistance profiling singly and in combination with the reference nucleoside inhibitor (aim 2). Allosteric candidates suitable for RdRp- targeted combination therapy will be synthetically optimized guided by ADME and pharmacokinetic profiles, followed by animal efficacy and toxicity assessment and synergy profiling in cell culture and in vivo (aim 3).

这项项目的总体假设是对人类的长期治疗成功 parainfluenzavirus 3型（HPIV3）和呼吸综合病毒（RSV）将需要组合疗法 与一对具有不同机械特征的病原体定向抑制剂接近。这个概念是由 大多数儿科患者人群要求的严格安全性和阻力威胁 针对个体疗法的突变可能会在循环病毒菌株中迅速固定。成员 HPIV3和RSV密切相关的Paramyxo-和肺炎病毒家族是大多数 美国婴儿的严重下呼吸道感染（LRI）和病毒疾病死亡，以及 被认为是对免疫功能低下和老年人的潜在威胁。两种病原体感染 在上呼吸道上皮启动，然后在患者中逐渐发展为小气道 前进到严重疾病，为治疗干预打开窗口。无疫苗保护或有效 目前可针对HPIV3或RSV提供治疗，以及针对RSV的抗体免疫预防 仅限于一部分高危患者。该项目将通过开发来满足这种未满足的临床需求 适用的，具有成本效益的治疗剂针对病毒RNA依赖性RNA聚合酶（RDRP）复合物。 在建立的抗病毒计划的基础上，我们最近确定了有效的核苷类似物 抑制剂具有对RSV和HPIV3的有效活性。作为参考，该化合物将告知 对变构的RDRP阻滞剂的共同开发，这些阻滞剂是从早期开发中严格审查的 与竞争性聚合酶抑制剂组合治疗的潜力。在试点研究中，我们设计了 重组HPIV3记者病毒，并在高通量筛查（HTS）广告系列中鉴定 将两个新型的新型HPIV3聚合酶抑制剂支架带有亚微摩尔摩尔摩尔起始效力。反对 RSV RDRP，我们已经重新设计了有效的变构麻疹病毒聚合酶阻滞剂和 在由此产生的药效团图书馆中确定了一个有效的新进入点，抗RSV RDRP 活动。认识到药物开发早期失败的风险，我们同时确定了 RSV聚合酶和封装基因组之间的蛋白质 - 蛋白质界面是一种有前途的 不充实的可药物位置。为了进一步多样化投资组合，也将变构抗RSV候选者多样化，本网站 将通过创新的生化HTS测定法和正交柜台审问（AIM 1）。这 现有的抗RSV和抗HPIV3线索以及新兴的候选人将是机械特征的 使用基于下一代的细胞和体外聚合酶测定，并进行抗性分析 并结合参考核苷抑制剂（AIM 2）。适用于RDRP-的变构候选者 有针对性的组合疗法将以ADME和药代动力学特征的指导进行合成优化， 然后进行动物疗效，毒性评估和在体内的协同分析（AIM 3）。

项目成果

Promotion of virus assembly and organization by the measles virus matrix protein.

• DOI: 10.1038/s41467-018-04058-2
• 发表时间: 2018-04-30
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Ke Z;Strauss JD;Hampton CM;Brindley MA;Dillard RS;Leon F;Lamb KM;Plemper RK;Wright ER
• 通讯作者: Wright ER

Identification and Characterization of Influenza Virus Entry Inhibitors through Dual Myxovirus High-Throughput Screening.

通过双粘病毒高通量筛选鉴定和表征流感病毒进入抑制剂。

• DOI: 10.1128/jvi.00898-16
• 发表时间: 2016
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: Weisshaar,Marco;Cox,Robert;Morehouse,Zachary;Kyasa,ShivaK;Yan,Dan;Oberacker,Phil;Mao,Shuli;Golden,JenniferE;Lowen,AniceC;Natchus,MichaelG;Plemper,RichardK
• 通讯作者: Plemper,RichardK

Envelope protein dynamics in paramyxovirus entry.

• DOI: 10.1128/mbio.00413-13
• 发表时间: 2013-07-02
• 期刊: mBio
• 影响因子: 6.4
• 作者: Plattet P;Plemper RK
• 通讯作者: Plemper RK

A stabilized respiratory syncytial virus reverse genetics system amenable to recombination-mediated mutagenesis.

• DOI: 10.1016/j.virol.2012.09.022
• 发表时间: 2012-12-05
• 期刊: Virology
• 影响因子: 3.7
• 作者: Hotard AL;Shaikh FY;Lee S;Yan D;Teng MN;Plemper RK;Crowe JE Jr;Moore ML
• 通讯作者: Moore ML

Structure and organization of paramyxovirus particles.

• DOI: 10.1016/j.coviro.2017.05.004
• 发表时间: 2017-06
• 期刊: Current opinion in virology
• 影响因子: 5.9
• 作者: Cox RM;Plemper RK
• 通讯作者: Plemper RK