Evaluation of Self-delivering FANA Antisense Oligonucleotide Lead Compounds for HIV Therapy

自递送 FANA 反义寡核苷酸先导化合物用于 HIV 治疗的评价

• 批准号: 10232090
• 负责人: Veenu Aishwarya
• 金额: $ 30万
• 依托单位: AUM LIFETECH, INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-07 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10232090
• 关键词: 13 year old; 5' Untranslated Regions; ADME Study; Acids; Acquired Immunodeficiency Syndrome; Acute; Adopted; Affect; Animals; Anions; Antisense Oligonucleotide Therapy; Antisense Oligonucleotides; Arabinose; Award; Base Pairing; Binding; Binding Proteins; Blood Cells; Blood specimen; CD8B1 gene; Cardiovascular Diseases; Caring; Cells; Centers for Disease Control and Prevention (U.S.); Chemistry; Chronic; Cleaved cell; Clinic; Data; Development; Dimerization; Dose; Drug Kinetics; Drug resistance; Endotoxins; Ensure; Excretory function; Funding; Future; Genome; Goals; HIV; HIV Infections; HIV therapy; HIV-1; Hepatocyte; High Pressure Liquid Chromatography; Hour; Human; Immune response; In Vitro; Infection; Lead; Legal patent; Malignant Neoplasms; Mediating; Metabolic; Metabolism; Modeling; Molecular Conformation; Mus; Mutate; Mutation; Outcome; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Pharmacology and Toxicology; Phase; Plasma; Plasma Proteins; Premature aging syndrome; Prevention; Process; Property; Publishing; RNA; RNA Interference; Rattus; Replacement Therapy; Resistance development; Ribonuclease H; Risk; Rodent; Safety; Self-Examination; Services; Signal Transduction; Site; Structure; Technology; Testing; Therapeutic; Therapeutic Effect; Toxic effect; Transcript; United States; Untranslated Regions; Viral; Viral Load result; Viral Packaging; Virus; Virus Diseases; Virus Replication; Widespread Disease; Withdrawal; Work; absorption; antiretroviral therapy; combinatorial; cytotoxic; cytotoxicity; design; dimer; drug candidate; effective therapy; genotoxicity; human DNA; humanized mouse; in vivo; metabolic abnormality assessment; mouse model; next generation; novel; pharmacokinetics and pharmacodynamics; prevent; prophylactic; reactivation from latency; ribonuclease H1; side effect; standard care; success; viral RNA; viral rebound

Project Summary/Abstract: Human immunodeficiency virus 1 (HIV-1) is the primary cause of acquired immune deficiency syndrome (AIDS) that affects over a million people in the United States alone. Advances in treatment have significantly prolonged the lives of those infected with HIV-1, with combinatorial antiretroviral therapy (cART) being the current standard in therapeutic care. However, cART alone cannot achieve complete eradication of the virus. Besides, drawbacks such as drug resistance development and severe side effects (e.g. premature aging, cancer, and cardiovascular disease) remain critical issues in cART therapy. Therefore, there is a need for a treatment, with efficient delivery and a favorable safety profile, that can reduce the HIV-1 viral load to undetectable levels. A promising approach is to utilize RNA silencing to treat HIV-1 infection by targeting the dimerization initiation site (DIS), a replication signal in the 5’ untranslated region (5’-UTR). Dimerization is initiated when the 5'-UTR undergoes a conformational change, allowing the DIS loops of two RNA genomes to base pair. This forms a kissing-loop (KL) dimer, which then leads to the subsequent packaging of the viral RNA. It is known that mutation or inhibition of the DIS severely restricts viral infectivity. Here we propose to test if 2’-deoxy-2’-fluoro-D-arabinonucleic acid antisense oligonucleotides (FANA ASOs), targeting DIS, will prevent viral replication. FANA ASOs will be designed to cleave (RNase H-dependent) or block viral RNA, thus inhibiting viral replication post-exposure. In a preliminary study using HIV-1 infected human peripheral blood mononuclear cells (PBMCs), DIS-targeting FANAs inhibited HIV-1 replication for as long as two weeks, after single doses of 400 nM and 3 µM doses (IC50 = 200 nM). FANA ASOs were also tested as prophylactics in vitro, at significantly lower concentrations, to prevent viral infection and amplification for up to 2 weeks. FANA ASOs were also designed to bind to DIS RNA without cleaving it, which would prevent the formation of the kissing loop structure that is necessary for replication. AUM- DIS-G9 emerged as the lead compound from our RNase H-dependent in vitro studies, while AUM-DIS-G0 will be the lead compound for our RNase H-independent trials. In this study, we will adopt a systematic approach to design and assess these two FANA lead ASOs targeting DIS. Our first aim is to evaluate the reduction in viral replication after treating with our two lead compounds, from our preliminary data, in a humanized NSG mouse model susceptible to HIV-1 infection. The second aim is to assess the potential of FANA ASOs as cART replacement in vivo, considering their prophylactic success in vitro, in a latently infected humanized mouse model. The third Aim is to perform standard pharmacokinetic and ADME studies on the lead compound from Aims 1 and 2 in vivo assessments. The goal of the proposed study is the development of a next generation antisense therapy for HIV.

项目概要/摘要： 人类免疫缺陷病毒1型（HIV-1）是获得性免疫缺陷综合征（艾滋病）的主要原因 仅在美国就有超过一百万人受到影响。治疗的进步显着延长了 HIV-1感染者的生命，联合抗逆转录病毒疗法（cART）是目前的标准 在治疗护理中。然而，单靠cART无法完全根除病毒。此外，缺点 如耐药性发展和严重的副作用（如过早衰老、癌症和心血管疾病）。 疾病）仍然是cART治疗中的关键问题。因此，需要一种治疗方法， 和良好的安全性，可以将HIV-1病毒载量降低到无法检测的水平。一种很有前途的方法 利用RNA沉默通过靶向二聚化起始位点（DIS）来治疗HIV-1感染， 在5'非翻译区（5'-UTR）中的信号。当5 '-UTR经历二聚化时，二聚化开始。 构象变化，允许两个RNA基因组的DIS环进行碱基配对。这就形成了接吻环（KL） 二聚体，然后导致病毒RNA的后续包装。已知突变或抑制 DIS严格限制了病毒的传染性。在这里，我们建议测试2 '-脱氧-2'-氟-D-阿拉伯核酸是否 靶向DIS的反义寡核苷酸（FANA ASO）将阻止病毒复制。FANA ASO将在 设计用于切割（RNase H依赖性）或阻断病毒RNA，从而抑制暴露后的病毒复制。中 使用HIV-1感染的人外周血单核细胞（PBMCs）、DIS靶向的初步研究 在400 nM和3 µM剂量单次给药后，FANA可抑制HIV-1复制长达两周（IC 50 = 200 nM）。FANA ASO还在体外以显著较低的浓度作为促炎剂进行了测试，以防止 病毒感染和扩增长达2周。FANA ASO还被设计为结合DIS RNA，而不 使其分裂，这将阻止复制所必需的吻环结构的形成。AUM- DIS-G9在我们的RNase H依赖性体外研究中作为先导化合物出现，而AUM-DIS-G 0将 作为我们的核糖核酸酶H独立试验的先导化合物在这项研究中，我们将采取系统的方法， 设计和评估这两种FANA先导ASO靶向DIS。我们的首要目标是评估病毒感染的减少， 根据我们的初步数据，在人源化NSG小鼠中用我们的两种先导化合物处理后， 模型易受HIV-1感染。第二个目标是评估FANA ASO作为cART的潜力 考虑到它们在体外的预防成功，在潜伏感染的人源化小鼠中进行体内替换 模型第三个目的是对来自以下的先导化合物进行标准药代动力学和ADME研究： 目的1和2体内评估。拟议研究的目标是开发下一代 HIV反义疗法