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

描述（由申请人提供）：抗OLAM适体作为新型镇痛药的开发（第2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。TRPV 1是瞬时受体电位（TRP）配体门控离子通道家族的重要成员，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证明了TRPV 1在炎性热痛觉过敏和其他疼痛疾病中的关键作用。氧化亚油酸代谢物（OLAMs）最近已被证明是一个新的家族内源性TRPV 1激动剂，有助于急性和炎性疼痛的条件。因此，阻断OLAM系统的化合物可能构成镇痛药的新家族。直接支持该预测，本文提供的1期数据证明，注射针对两种主要OLAM：9-HODE和13-HODE开发的高亲和力DNA适体在热痛的膜片钳和大鼠行为模型中产生显著的镇痛作用.虽然这些数据为概念验证提供了证据，但适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，Operational Technologies Corporation（OpTech）提出继续将其成功的高亲和力抗OLAM适体从阶段1开发成体内长寿命适体缀合物，其特异性结合9-HODE和13-HODE并中和它们的疼痛产生活性持续延长的时间段。这将允许用更特异、更便宜和更高亲和力的DNA适体代替抗HODE多克隆抗体。在II期研究中，OpTech预计将通过在3'端添加布洛芬（参考文献80），从而实现几个旨在增强药代动力学（PK）的特定目标，从而能够与血清白蛋白结合，以增加体重并延迟肾脏和肝脏清除。还将研究减缓体内清除和保护体内适体的替代但经证实的方法，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相互作用的三维分子模型，以实现对适体亲和力（已经表现出低nM KD值）和特异性的潜在分子工程改进，如果可能的话，使用修饰的或非天然的脱氧核苷酸。除了在存在和不存在各种剂量的适体缀合物的情况下的体外膜片钳研究之外，还将研究大鼠在注射和暴露于43 ° C有害热之前和之后的行为变化以及各种适体-3 '-缀合物。如果观察到相对长期（小时量级）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。

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.