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）：Anti-OLAM适配体作为新型镇痛药的开发（第二阶段）由于对疼痛机制的不完全理解，疼痛的管理仍然是一个主要的卫生保健问题。TRPV1是瞬时受体电位（TRP）家族配体门控离子通道的重要成员，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都表明，TRPV1在炎症性热痛觉过敏和其他疼痛状况中起着关键作用。氧化亚油酸代谢物（OLAMs）最近被证明包含一个新的内源性TRPV1激动剂家族，有助于急性和炎症性疼痛。因此，阻断OLAM系统的化合物可能构成一个新的镇痛药家族。直接支持这一预测的是，本文提供的1期数据表明，注射针对两种主要OLAMs （9-HODE和13-HODE）的高亲和力DNA适配体，在膜片钳和大鼠热痛行为模型中产生了显著的镇痛作用。虽然这些数据为概念验证提供了证据，但为了避免肾脏和其他主要器官的快速清除，必须将适体发展成更重的缀合物。因此，Operational Technologies Corporation （OpTech）建议继续将其成功的高亲和力抗olam适配体从第一阶段开发成体内长寿命的适配体偶联物，特异性结合9-HODE和13-HODE，并在较长时间内中和它们产生疼痛的活性。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。在第二阶段，OpTech希望通过在3'端添加布洛芬来完成几个针对增强药代动力学（PK）的特定目标（参考文献80），从而使其与血清白蛋白结合，从而增加重量并延缓肾脏和肝脏的清除。此外，还将研究减缓体内清除和保护适体（包括3‘-聚乙二醇（PEG）和共价3’大鼠白蛋白附着）的替代方法。此外，西南研究所（SwRI）将生成适配体- hode相互作用的三维分子模型，以便在可能的情况下，使用修饰或非天然脱氧核苷酸，对适配体亲和性（已经表现出较低的nM KD值）和特异性进行潜在的分子工程改进。除了在体外膜片钳中研究不同剂量适配体偶联物存在和不存在的情况外，还将研究大鼠在注射和暴露于43℃有毒高温前后与各种适配体-3'偶联物的行为变化。如果观察到相对长期（大约几小时）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pre-IND申请。