Development of anti-sense oligonucleotides as a therapeutic for cancer pain through selective block of sodium channel pain targets

开发反义寡核苷酸通过选择性阻断钠通道疼痛靶标来治疗癌症疼痛

• 批准号: 10081895
• 负责人: Owen B. McManus
• 金额: $ 30.07万
• 依托单位: QUELL TX, INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-07 至 2021-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10081895
• 关键词: Afferent Neurons; Animal Model; Antisense Oligonucleotides; Binding; Biological Assay; Breast Cancer Patient; Cancer Model; Cancer Patient; Cell model; Cells; Chemistry; Clinic; Coupled; Data; Development; Disease; Dissociation; Dose; Drug Kinetics; Electrophysiology (science); Estrogen receptor positive; Evaluation; Funding; Goals; Human; Human Activities; Immune response; In Vitro; Individual; Inflammation Mediators; Injections; Laboratories; Lead; Licensing; Life Expectancy; Malignant Neoplasms; Measures; Mediator of activation protein; Medical; Messenger RNA; Metastatic Neoplasm to the Bone; Modality; Monkeys; Neurons; Nociception; Opioid; Optics; Pain; Patients; Peripheral; Pharmacodynamics; Phase; Phenotype; Physiological; Preparation; Proteins; Protocols documentation; RNA; RNA Degradation; Rattus; Research; Rivers; Route; Sampling; Sensory; Small Business Innovation Research Grant; Sodium Channel; Spinal Ganglia; System; Technology; Testing; Therapeutic; Tissues; Toxic effect; Transcript; Treatment Efficacy; Vertebral column; addiction; base; cancer pain; channel blockers; design; effective therapy; efficacy study; efficacy testing; end of life; immunocytochemistry; in vivo; knock-down; mRNA Expression; nervous system disorder; neuronal excitability; novel; pain model; pharmacokinetics and pharmacodynamics; response; selective expression; side effect; single-cell RNA sequencing; small molecule; standard of care; temporal measurement; transmission process; tumor; voltage

Project Summary: Development of anti-sense oligonucleotides as a therapeutic for cancer pain by selectively reducing sodium channel expression Effective treatment of cancer pain is a large unmet medical need, as opioids, the current standard of care lack efficacy and cause addiction. For end-of-life cancer patients and estrogen receptor- positive breast cancer patients with long life expectancy, more effective therapeutic approaches for pain are needed. Antisense oligonucleotides (ASOs) bind to and induce degradation of RNA transcripts with specific sequences, enabling selective protein knockdown (KD) with long duration of action (weeks or months), to deliver long-term relief. ASOs can also be used in combination, to reduce the expression of multiple proteins for increased efficacy. As a modality, ASOs have been successfully applied in the clinic to treat severe neurological disease. We target the voltage gated sodium channels (Navs) selectively expressed in dorsal root ganglion neurons (Nav1.7, Nav1.8 and Nav1.9), which are implicated in pain transmission and specifically in cancer pain. Reduction of channel expression aims to overcome the limitations of small molecule state-dependent channel blockers, if the channel should be blocked in a state-independent manner for efficacy. Preliminary results with our designed ASOs show >70% mRNA KD for several Navs in rat dorsal root ganglion sensory neurons (DRGs). We apply a novel platform to design ASOs for specific knockdown of Nav channels. For pain targets and indications, QuellTx also has an exclusive worldwide license to breakthrough technology: (i) An in vitro cellular model for cancer pain (developed through SBIR funding by NCI), where primary DRG neurons are bathed in a physiologically-relevant mixture of inflammatory mediators secreted by tumors, or ‘cancer-SPARC’. The SPARC- treated neurons become hyperexcitable, mimicking the cellular pain response. (ii) An all optical electrophysiology high throughput readout for neuronal excitability (Optopatch), allowing recordings from 100s of individual neurons in parallel with high temporal resolution. This platform enables us to determine the impact of modulating expression of Nav1.7, 1.8 and 1.9, alone or in combination, on the pain-in-a-dish phenotype. In Phase I we propose to leverage our ASO design capabilities, combined with the breakthrough technology to: (a) identify ASOs for selective modulation of expression of Nav channel pain targets and (b) determine the combination of ASOs and %mRNA knock down for maximal phenotypic effect in vitro. We will select ASOs that efficiently knockdown Nav channels and reverse the pain phenotype induced by cancer-SPARC. In Phase II we will determine the PK/PD (%mRNA KD) in rats, and the % mRNA KD needed for efficacy in a rat model of cancer pain, while optimizing human cross-reacting with cynmolgous monkey (cyno) ASOs for efficacy in human DRG neuronal models of pain. Our ultimate goal is to advance optimized ASOs to IND enabling toxicity studies in rat and cynos, leading to a therapeutic to treat cancer pain in patients.

项目概述：开发反义寡核苷酸作为癌症疼痛的治疗药物， 选择性降低钠通道表达 有效治疗癌痛是一个很大的未满足的医疗需求，作为阿片类药物，目前的护理标准缺乏 功效和致瘾性。对于晚期癌症患者和雌激素受体阳性乳腺癌 对于预期寿命较长的患者，需要更有效的疼痛治疗方法。反义 寡核苷酸（ASO）结合并诱导具有特定序列的RNA转录物的降解， 选择性蛋白质敲除（KD），作用持续时间长（数周或数月），可提供长期缓解。 ASO也可以组合使用，以减少多种蛋白质的表达，从而提高功效。作为 作为一种形式，ASO已成功地应用于临床治疗严重的神经系统疾病。 我们靶向背根神经节神经元选择性表达的电压门控钠通道（Navs （Nav1.7、Nav1.8和Nav1.9），其涉及疼痛传递，特别是癌症疼痛。 减少通道表达的目的是克服小分子状态依赖性通道的局限性 阻断剂，如果通道应以状态无关的方式阻断以获得疗效。初步结果与 我们设计的ASO在大鼠背根神经节感觉神经元（DRG）中对几种Navs显示> 70%mRNA KD。 我们应用一种新的平台来设计用于特定敲除Nav通道的ASO。针对疼痛目标， QuellTx还拥有突破性技术的全球独家许可证：（i）体外细胞 癌症疼痛模型（由NCI通过SBIR资助开发），其中初级DRG神经元沐浴在一个 肿瘤是由肿瘤分泌的炎性介质的生理相关混合物，或“癌症-SPARC”。这是... 被处理的神经元变得过度兴奋，模仿细胞疼痛反应。(ii)全光学电生理学 神经元兴奋性的高通量读出（Optopatch），允许记录100个单个神经元 同时具有高时间分辨率。该平台使我们能够确定调制的影响 Nav1.7、1.8和1.9单独或组合在培养皿疼痛表型上的表达。 在第一阶段，我们建议利用我们的阿索设计能力，结合突破性技术： (a)鉴定用于选择性调节Nav通道疼痛靶标的表达的ASO，和（B）确定 AS 0和%mRNA敲低的组合用于体外最大表型效应。我们将选择符合以下条件的ASO： 有效敲除Nav通道并逆转癌症-SPARC诱导的疼痛表型。在第二阶段，我们 将确定大鼠中的PK/PD（%mRNA KD），以及在大鼠癌症模型中有效性所需的% mRNA KD 疼痛，同时优化人与食蟹猴（cyno）ASO的交叉反应，以获得人DRG中的疗效 疼痛的神经元模型我们的最终目标是将优化的ASO推进到IND，以实现大鼠毒性研究 和cynos，导致治疗癌症疼痛的患者的治疗。