Development of Anti-OLAM Aptamers as Novel Analgesics

描述（由申请人提供）：抗olam适体作为新型镇痛药的发展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，本文提供的初步数据表明，注射针对两种主要OLAMs （9-HODE和13-HODE）的抗体，在两种疼痛模型中产生显著的镇痛作用。尽管这些数据为概念验证提供了证据，但由于与免疫原性相关的不良反应，兔多克隆抗体的临床开发是不可行的。因此，Operational Technologies Corporation （OpTech）建议使用组合适体化学来发现特异性结合这些olam并中和其产生疼痛活动的DNA适体。这将允许用更特异、更便宜、可能更高亲和力的DNA适体取代抗体。在第一阶段，OpTech希望完成两个总体的具体目标。Aim 1将开发、克隆和测序几种高度特异性的DNA适体，这些适体结合9-羟基十二烯酸（HODE）和13-HODE，而不结合前体脂质亚油酸（图1A）。目的2将使用几种体内临床前大鼠疼痛模型来评估适配体的镇痛活性。在第二阶段，OpTech将进行细化、优化和启动

DESCRIPTION (provided by applicant): Development of Anti-OLAM Aptamers as Novel Analgesics 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 channel, 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 contributes 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, preliminary data provided herein demonstrate that injection of antibodies against two of the major OLAMs, 9-HODE and 13-HODE, produce significant analgesia in two pain models. Although these data provide evidence for proof-of-concept, the clinical development of rabbit polyclonal antibodies is not feasible due to adverse effects related to immunogenicity. Accordingly, Operational Technologies Corporation (OpTech) proposes to use combinatorial aptamer chemistry to discover DNA aptamers that specifically bind to these OLAMs and neutralize their pain-producing activities. This would permit replacing antibodies with more specific, less expensive and perhaps higher affinity DNA aptamers. In Phase 1, OpTech expects to complete two overall Specific Aims. Aim 1 will develop, clone, and sequence several highly specific DNA aptamers that bind 9-hydroxydecadienoic acid (HODE) and 13-HODE without binding to the precursor lipid, linoleic acid (Fig 1A). Aim 2 will evaluate the analgesic activity of the aptamers using several in vivo preclinical rat models of pain. In Phase 2, OpTech will refine, optimize, and begin commercialization of its anti-OLAM aptamer compounds. The Phase 2 optimization process will include 3-D modeling of putative aptamer binding pocket interactions with the OLAMs. Based on 3-D modeling findings, OpTech anticipates adding modified bases having various functional groups (e.g., primary amines, methyl, thiol groups, etc.) that are now commercially available for addition to oligonucleotides at the point of chemical synthesis. The effects of these additional functional groups on aptamer-OLAM binding affinity are expected to better emulate amino acid side chains and will be studied by ELISA-like plate assays and surface plasmon resonance (SPR). The highest affinity and most specific unmodified or modified anti-OLAM aptamers will move into animal studies. The most effective aptamers in animal pain studies will be modified for longer in vivo lifetimes (3'-cholesterol addition and inclusion in liposomes, PEGylation, etc.) or time-released formulation and enter the FDA approval pipeline. PUBLIC HEALTH RELEVANCE: Development of Anti-OLAM Aptamers as Novel Analgesics Millions of patients suffer from pain and many available analgesic drugs ("pain killers") suffer from either incomplete analgesia or unacceptable side-effects. The proposal will develop a novel class of analgesics that work by blocking the endogenous capsaicin-like substances that are released during tissue injury.