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: 10011742
• 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/10011742
• 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.

项目摘要/摘要： 人免疫缺陷病毒1（HIV-1）是获得免疫缺陷综合征（AIDS）的主要原因 仅在美国就影响了超过一百万的人。治疗的进展显着延长 感染HIV-1的人的生活，结合抗逆转录病毒疗法（CART）是当前标准 在理论护理中。但是，单独的推车无法完全消除病毒。此外，缺点 例如耐药性发展和严重的副作用（例如，早衰，癌症和心血管 疾病）在购物车治疗中仍然是关键问题。因此，需要治疗，有效地交付 以及有利的安全性，可以将HIV-1病毒载荷降低到无法检测到的水平。一种有希望的方法 是利用RNA沉默来通过靶向二聚化启动部位（DIS）来治疗HIV-1感染 在5'未翻译区域（5'-UTR）中发出信号。当5'-UTR经历A时启动二聚化 构象变化，使两个RNA基因组的循环得出碱基对。这形成一个接吻环（KL） 二聚体，然后导致病毒RNA的随后包装。众所周知，突变或抑制 这些严重限制了病毒感染。在这里，我们建议测试2'-Deoxy-2'-氟-d- d-阿拉伯核酸 反义寡核苷酸（FANA ASOS）靶向DIS将防止病毒复制。 fana asos将是 旨在清除（RNase H依赖性）或阻断病毒RNA，从而抑制暴露后病毒复制。在 使用HIV-1感染人类外周血单核细胞（PBMC）的初步研究，靶向 FANAS抑制HIV-1复制长达两周，在单剂量400 nm和3 µm剂量之后（IC50） = 200 nm）。 FANA ASO还以明显较低的浓度在体外测试了预防药物，以防止 病毒感染和扩增长达2周。 Fana Asos也被设计为与Dis rna结合而没有 切割它，这将防止复制所需的接吻环结构的形成。 aum- DIS-G9从我们的RNase H依赖性研究中作为铅化合物出现，而AUM-DIS-G0将 成为我们RNase H非依赖性试验的铅化合物。在这项研究中，我们将采用系统的方法 设计和评估这两个FANA LEAD ASOS靶向DIS。我们的第一个目的是评估病毒的减少 从我们的初步数据中使用我们的两种铅化合物处理后的复制，在人源化的NSG小鼠中 容易受到HIV-1感染的模型。第二个目的是评估FANA ASOS AS CART的潜力 在体外的预防性成功中，在体外取代，在潜在感染的人源性小鼠中 模型。第三个目的是对铅化合物进行标准的药代动力学和ADME研究 目标1和2体内评估。拟议的研究的目的是发展下一代 艾滋病毒的反义治疗。