Development of Anti-OLAM Aptamers as Novel Analgesics (Phase 2)

开发抗 OLAM 适体作为新型镇痛药（第 2 阶段）

• 批准号: 9059113
• 负责人: John G Bruno
• 金额: $ 46.5万
• 依托单位: OPERATIONAL TECHNOLOGIES CORPORATION
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9059113
• 关键词: 3-Dimensional; Absence of pain sensation; Acids; Acute inflammatory pain; Adverse effects; Afferent Neurons; Affinity; Agonist; Albumins; American; Analgesics; Animals; Antibodies; Arthritis; Behavioral Model; Binding; Burn injury; Burning Pain; Capsaicin; Chemicals; Clinical; Coronary heart disease; DNA; Data; Development; Diabetes Mellitus; Diagnosis; Dose; Drug Kinetics; Engineering; Evolution; Exhibits; Exposure to; FDA approved; Family; Foundations; Gated Ion Channel; Gene Deletion; Health; Healthcare; Heart; Heating; Hepatic; Hour; Hyperalgesia; Ibuprofen; In Vitro; Incidence; Inflammatory; Injectable; Injection of therapeutic agent; Injury; Investigational Drugs; Investigational New Drug Application; Kidney; Life; Ligands; Linoleic Acids; Liver; Malignant Neoplasms; Mechanics; Mediating; Medical; Metabolite Interaction; Methods; Modeling; Molecular; Molecular Models; Nociceptors; Nucleic Acids; Nucleotides; Opioid; Organ; Oryctolagus cuniculus; Pain; Pain management; Patients; Peripheral; Pharmaceutical Preparations; Phase; Physiological; Polyethylene Glycols; Process; Proteins; Quality of life; Rattus; Reporting; Research Institute; Role; Serum Albumin; Shock; Small Business Innovation Research Grant; Sodium; Specificity; Stimulus; Stroke; System; TRPV1 gene; Technology; Testing; Time; Tissues; Ulcer; Weight; Work; addiction; aptamer; base; behavioral study; cancer pain; chronic pain; clinical investigation; cost; design; immunogenic; immunogenicity; in vivo; inhibiting antibody; injured; member; molecular modeling; novel; pain inhibition; patch clamp; pegaptanib; polyclonal antibody; receptor

DESCRIPTION (provided by applicant): Development of Anti-OLAM Aptamers as Novel Analgesics (Phase 2) The management of pain remains a major health care problem due to an incomplete understanding of pain mechanisms. TRPV1, a prominent member of the transient receptor potential (TRP) family of ligand-gated ion channels, detects noxious chemical and physical stimuli in peripheral tissues. Both pharmacological and gene deletion studies have demonstrated a pivotal role for TRPV1 in inflammatory heat hyperalgesia and other pain conditions. Oxidized linoleic acid metabolites (OLAMs) have been recently demonstrated to comprise a novel family of endogenous TRPV1 agonists that contribute to acute and inflammatory pain conditions. Therefore, compounds that block the OLAM system are likely to constitute a novel family of analgesics. In direct support of this prediction, Phase 1 data provided herein demonstrate that injection of high affinity DNA aptamers developed against two of the major OLAMs: 9-HODE and 13-HODE, produced significant analgesia in patch- clamp and rat behavioral models of heat pain. Although these data provide evidence for proof-of-concept, the aptamers must be developed into heavier conjugates in order to avoid rapid clearance by the kidneys and other major organs. Accordingly, Operational Technologies Corporation (OpTech) proposes to continue developing its successful high affinity anti-OLAM aptamers from Phase 1 into long-lived aptamer conjugates in vivo which specifically bind to 9-HODE and 13-HODE and neutralize their pain-producing activities for extended periods of time. This would permit replacing anti-HODE polyclonal antibodies with more specific, less expensive and higher affinity DNA aptamers. In Phase 2, OpTech expects to complete several specific aims targeted toward enhancing pharmacokinetics (PK) by addition of ibuprofen to the 3' end (ref. 80), thereby enabling association with serum albumin to add weight and retard renal and hepatic clearance. Alternative, but proven, methods to slow in vivo clearance and protect aptamers in vivo including 3'-polyethylene glycol (PEG) and covalent 3' rat albumin attachment will also be investigated. Moreover, 3-dimensional molecular models of aptamer-HODE interactions will be generated by the Southwest Research Institute (SwRI) to enable potential molecular engineering of improvements to aptamer affinity (already exhibiting low nM KD values) and specificity, if possible, using modified or unnatural deoxynucleotides. In addition to in vitro patch clamp studies in the presence and absence of various doses of the aptamer conjugates, rats will be studied for behavioral changes before and after injection and exposure to 43oC noxious heat with the various aptamer-3'-conjugates. If relatively long- term (on the order of hours) PK is observed and heat/burn analgesia can again be demonstrated, OpTech will seek to file an IND or pre-IND application with the FDA.

描述（由申请人提供）：作为新型镇痛药的抗 OLAM 适体的开发（第 2 阶段） 由于对疼痛机制的不完全了解，疼痛的管理仍然是一个主要的卫生保健问题。 TRPV1 是配体门控离子通道瞬时受体电位 (TRP) 家族的重要成员，可检测外周组织中的有害化学和物理刺激。药理学和基因缺失研究均证明 TRPV1 在炎性热痛觉过敏和其他疼痛病症中发挥着关键作用。最近已证明氧化亚油酸代谢物 (OLAM) 包含一个新的内源性 TRPV1 激动剂家族，可导致急性和炎症性疼痛。因此，阻断 OLAM 系统的化合物可能构成一个新的镇痛药家族。为了直接支持这一预测，本文提供的第一阶段数据表明，注射针对两种主要 OLAM（9-HODE 和 13-HODE）开发的高亲和力 DNA 适体，在膜片钳和热痛大鼠行为模型中产生显着的镇痛作用。尽管这些数据为概念验证提供了证据，但适配体必须开发成更重的缀合物，以避免被肾脏和其他主要器官快速清除。因此，OpTech 公司建议继续将第一阶段成功的高亲和力抗 OLAM 适体开发成体内长寿命的适体缀合物，该缀合物特异性结合 9-HODE 和 13-HODE 并中和它们产生疼痛的活性较长的一段时间。这将允许用更特异、更便宜和更高亲和力的DNA适体替代抗HODE多克隆抗体。在第 2 阶段，OpTech 预计通过在 3' 末端添加布洛芬（参考文献 80）来完成几个旨在增强药代动力学 (PK) 的具体目标，从而能够与血清白蛋白结合以增加体重并延缓肾脏和肝脏清除。还将研究减缓体内清除和保护体内适体的替代但经过验证的方法，包括 3'-聚乙二醇 (PEG) 和共价 3' 大鼠白蛋白附着。此外，西南研究所（SwRI）将生成适体-HODE相互作用的3维分子模型，以实现潜在的分子工程改进适体亲和力（已经表现出低nM KD值）和特异性，如果可能的话，使用修饰或非天然脱氧核苷酸。除了在存在和不存在不同剂量的适体缀合物的情况下进行体外膜片钳研究之外，还将研究大鼠在注射各种适体-3'-缀合物之前和之后以及暴露于43oC有害热量之前和之后的行为变化。如果观察到相对长期（数小时）的 PK 并且可以再次证明热/烧伤镇痛，OpTech 将寻求向 FDA 提交 IND 或预 IND 申请。