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： 人类免疫缺陷病毒 1 (HIV-1) 是获得性免疫缺陷综合症 (AIDS) 的主要原因 仅在美国，这种疾病就影响了超过一百万人。 HIV-1 感染者的生活，组合抗逆转录病毒治疗 (cART) 是当前的标准 然而，仅靠 cART 并不能完全消灭病毒。 例如耐药性的发展和严重的副作用（例如过早衰老、癌症和心血管疾病） 疾病）仍然是 cART 治疗中的关键问题，因此，需要一种有效的治疗方法。 和良好的安全性，可以将 HIV-1 病毒载量降低到不可检测的水平，这是一种有前途的方法。 是利用RNA沉默来治疗HIV-1感染，通过靶向二聚化起始位点（DIS），复制 当 5'-UTR 发生变化时，5'非翻译区 (5'-UTR) 中的信号就会启动。 构象变化，使两个 RNA 基因组的 DIS 环碱基配对，形成接吻环 (KL)。 二聚体，然后导致病毒 RNA 的后续包装，已知突变或抑制。 DIS 严重限制了病毒的传染性。在此，我们建议测试 2'-脱氧-2'-氟-D-阿拉伯核酸。 针对 DIS 的反义寡核苷酸 (FANA ASO) 将阻止病毒复制。 旨在裂解（RNase H 依赖性）或阻断病毒 RNA，从而抑制病毒在暴露后的复制。 使用 HIV-1 感染的人外周血单核细胞 (PBMC) 进行的初步研究，DIS 靶向 单剂量 400 nM 和 3 µM 剂量后，FANA 可抑制 HIV-1 复制长达两周（IC50 = 200 nM) 还对 FANA ASO 作为体外预防剂进行了测试，浓度显着降低，以预防。 FANA ASO 还被设计为无需与 DIS RNA 结合即可进行长达 2 周的病毒感染和扩增。 切割它，这将阻止复制所需的接吻环结构的形成。 DIS-G9 成为我们的 RNase H 依赖性体外研究中的先导化合物，而 AUM-DIS-G0 将 是我们不依赖 RNase H 的试验的先导化合物。在这项研究中，我们将采用系统方法来进行研究。 设计和评估这两个针对 DIS 的 FANA 主要 ASO 我们的首要目标是评估病毒减少情况。 根据我们的初步数据，用我们的两种先导化合物处理后，在人源化 NSG 小鼠中进行复制 易受 HIV-1 感染的模型的第二个目标是评估 FANA ASO 作为 cART 的潜力。 考虑到它们在体外预防潜伏感染的人源化小鼠中的成功，体内替代 第三个目标是对先导化合物进行标准药代动力学和 ADME 研究。 目标 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。