Toxins from natural venoms modulate pain sensitization by depolarization-independent mechanisms in sensory neurons

目前的镇痛药，如非甾体抗炎药（NSAID）和阿片类药物往往导致超过20%的患者疼痛缓解不足。此外，目前的治疗剂如阿片类药物受到其副作用如便秘、恶心、呼吸抑制和成瘾的强烈限制1。新的细胞和分子疼痛机制的确定是开发新型镇痛药物的迫切需要。有毒生物进化出了具有高分子特异性和有效性的毒素混合物。许多毒素引起疼痛，而另一些则抑制疼痛。毒素在细胞和分子机制的鉴定以及药物开发方面的作用已经得到了很好的证实。然而，它们用于靶点识别和作为缓解疼痛的治疗化合物的用途仍然是一个新兴领域2。目前，最有效的镇痛药物如NSAID和阿片类药物调节GPCR信号传导，从而调节伤害性神经元的敏感性和活性。通过我们最近的研究，我们成功地证明，用我们创新的“高含量筛选（HCS）”显微镜方法监测细胞内信号能够检测致敏、脱敏和去极化诱导化合物。在我们目前资助的“毒液/毒素项目”中，我们已经确定了去极化诱导的致敏信号的成分。我们现在正着手从我们目前对去极化依赖性致敏毒素的研究中取得进展，以从天然毒液中鉴定出不依赖于去极化的亲伤害性和镇痛性化合物。为此，在科隆（C）和莫斯科（M），我们将追求以下目标：1）从各种毒液中鉴定新的去极化非依赖性致敏/镇痛毒素。测试毒液的细胞脱敏/镇痛活性（C，M）B。从毒液中分离单独活性毒素的生化分级分离和细胞测试（C，M）c.活性毒素的结构测定（M）2）作用的细胞和分子机制分析a.活性肽（M）的重组类似物的产生B.鉴定的毒素（C，M）的药理学表征（剂量反应，动力学）c.受活性毒素影响的伤害感受神经元的亚组特异性（C）d.细胞机制分析（C）3）治疗潜力分析a。我们在动物行为测试中的细胞和分子数据的机制和治疗确证（C，M）

Current analgesics such as nonsteroidal anti-inflammatory drugs (NSAIDs) and opioids often result in insufficient pain relief in more than 20% of patients. In addition, current therapeutics such as opioids are strongly limited by their side effects such as constipation, nausea, respiratory depression, and addiction1. Novel cellular and molecular pain mechanisms need to be defined urgently for the development of novel analgesic compounds.Venomous organisms evolved toxin mixtures of high molecular specificity and effectiveness. Many toxins evoke pain, and others inhibit pain. The power of toxins for the identification of cellular and molecular mechanisms as well as drug development is well established. Nevertheless, their use for target identification and as therapeutic compounds for pain relief is still an emerging field2. Currently, the most potent analgesic drugs such as NSAIDs and opioids modulate GPCR signaling and thereby the sensitivity and activity of nociceptive neurons. With our recent studies we succeeded to prove, that monitoring intracellular signaling with our innovative “High Content Screening (HCS)” microscopy approach is able to detect sensitization, desensitization, and depolarization inducing compounds. In our currently funded "venom/toxin Project" we have identified components for depolarization-induced sensitization signaling. We are now setting out to progress from our current research on depolarization-dependent sensitizing toxins, to identify depolarization-independent pro-nociceptive as well as analgesic compounds from natural venoms. For this, in Cologne (C) and Moscow (M) we will pursue the following objectives:1) Identification of novel depolarization-independent sensitizing/analgesic toxins from a variety of venomsa. Test venoms for cellular desensitizing/analgesic activity (C, M)b. Biochemical fractionation and cellular testing to isolate individual active toxins from the venoms (C, M)c. Structure determination of the active toxins (M)2) Analysis of the cellular and molecular mechanism of actiona. Production of recombinant analogs of the active peptides (M)b. Pharmacological characterization (dose-responses, kinetics) of the identified toxins (C, M)c. Subgroup specificity of nociceptive neurons affected by the active toxins (C)d. Analysis of the cellular mechanism (C)3) Analysis of therapeutic potentiala. Mechanistic and therapeutic corroboration of our cellular and molecular data in animal behavioral tests (C, M)