Intra-cartilage depot delivery of electrically-charged IL-1RA for targeting osteoarthritis-associated inflammation and catabolism in multiple joint tissues

软骨内储库递送带电 IL-1RA，用于靶向多个关节组织中与骨关节炎相关的炎症和分解代谢

基本信息

批准号：
10861426
负责人：
Ambika Goel Bajpayee
金额：
$ 42.26万
依托单位：
NORTHEASTERN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-21 至 2026-07-31
项目状态：
未结题

项目摘要

Project Summary/Abstract Osteoarthritis (OA) pain remains an intractable problem with few treatment options. Centrally acting analgesic drugs carry the risk of addictive side effects. Thus, local sustained analgesic delivery is an attractive strategy for development of novel OA pain therapies. Local intra-articular (IA) delivery, however, remains inadequate due to rapid drug clearance from the synovial joint space requiring multiple injections that cause toxicity. This is further complicated by the dense and highly negatively charged cartilage matrix that prevents drugs from reaching their target sites. The parent R01 addressed this challenge by developing cartilage targeting and residing cationic anti-catabolic and pro-anabolic disease modifying OA therapeutics. For example, a receptor antagonist of IL-1 (IL-1RA) was fused with an optimally charged cationic peptide carrier (CPC-IL-1RA), and its effectiveness in suppressing OA associated catabolism was demonstrated in-vitro as well as in vivo with only a one-time dose. Here we will broaden the impact of our work in targeted drug delivery to treatment of OA pain by evaluating the effectiveness in sustained release formulations of (i) cationic fusion CPC-IL-1RA (already developed as part of the parent R01 tasks) and (ii) cationic CCR2 receptor antagonist (CCR2RA). We and others have shown that CCL2/CCR2 signaling is key for promoting persistent pain in the destabilization of the medial meniscus (DMM) model and that its local pharmacological blockade decreases pain behavior. These findings suggest that CCL2- CCR2 signaling locally in the joint also contributes to pain in experimental OA, highlighting the need for developing sustained local delivery methods for CCR2 inhibitors such as CCR2 receptor antagonist (CCR2RA), which is a small molecule. CCR2RA will be fused with a cationic cartilage targeting nano-construct developed for delivery of small molecules, multi-arm Avidin (mAv). The central hypothesis is that a one-time IA injection of CPC-IL-1RA and mAv-CCR2RA will provide sustained analgesic relief over 4 weeks in a rat medial meniscal transection (MMT) OA model. We have a team with expertise in OA pain and in musculoskeletal joint drug delivery that will test this hypothesis in the following aims, Aim 1: Synthesize an intra-cartilage depot for a month- long sustained release of CCR2RA using cationic mAv using hydrolysable ester linkers. Drug release rates and bioactivity of the conjugate will be measured and compared with unmodified drug in-vitro. Aim 2: Test the efficacy of a single IA injection of cationic CPC-IL-1RA and mAv-CCR2RA in the rat MMT model. Male and female rats will receive an IA injection of saline, CPC-IL-1RA, unconjugated IL-1RA, mAv-CCR2RA, or unconjugated CCR2RA. Weight-bearing asymmetry and knee hyperalgesia will be assessed; DRG changes will be evaluated by IHC and RNAscope; joint histopathology will be assessed, including assessment of knee innervation changes and immune cell infiltration. This competitive urgent revision will broaden the impact of the already designed cationic therapeutics for sustained OA pain suppression over a month with only a single IA administration thereby providing long-term pain relief, in addition to disease modification (which was addressed in the original R01).

项目概要/摘要骨关节炎（OA）疼痛仍然是一个棘手的问题，治疗选择很少。中枢镇痛药药物具有令人上瘾的副作用的风险。因此，局部持续镇痛给药对于患者来说是一种有吸引力的策略。开发新型 OA 疼痛疗法。然而，由于以下原因，局部关节内（IA）递送仍然不足：药物从滑膜关节间隙快速清除，需要多次注射，从而引起毒性。这进一步由于致密且带高负电荷的软骨基质阻碍药物到达其作用，使情况变得复杂目标站点。母体 R01 通过开发软骨靶向和驻留阳离子来应对这一挑战抗分解代谢和促合成代谢疾病改善 OA 疗法。例如，IL-1受体拮抗剂 (IL-1RA) 与最佳电荷阳离子肽载体 (CPC-IL-1RA) 融合，及其在体外和体内均证明只需一次性剂量即可抑制 OA 相关的分解代谢。在这里，我们将通过评估 OA 疼痛的治疗，扩大我们在靶向药物递送方面的工作的影响。 (i) 阳离子融合 CPC-IL-1RA（已作为母体 R01 任务）和 (ii) 阳离子 CCR2 受体拮抗剂 (CCR2RA)。我们和其他人已经证明 CCL2/CCR2 信号传导是促进内侧半月板 (DMM) 不稳定中持续疼痛的关键模型及其局部药理学阻断可减少疼痛行为。这些发现表明 CCL2- 关节中局部的 CCR2 信号传导也会导致实验性 OA 的疼痛，这突出表明需要开发 CCR2 抑制剂（例如 CCR2 受体拮抗剂 (CCR2RA)）的持续局部递送方法，这是一个小分子。 CCR2RA 将与开发的靶向纳米结构的阳离子软骨融合用于输送小分子、多臂亲和素 (mAv)。中心假设是一次性 IA 注射 CPC-IL-1RA 和 mAv-CCR2RA 将为大鼠内侧半月板提供超过 4 周的持续镇痛缓解横断面 (MMT) OA 模型。我们拥有一支在骨关节炎疼痛和肌肉骨骼关节药物方面具有专业知识的团队交付将在以下目标中测试这一假设，目标 1：合成一个月的软骨内库 - 使用可水解酯连接体的阳离子 mAv 长时间持续释放 CCR2RA。药物释放速率和将测量缀合物的生物活性并与体外未修饰的药物进行比较。目标2：测试功效在大鼠 MMT 模型中单次 IA 注射阳离子 CPC-IL-1RA 和 mAv-CCR2RA。雄性和雌性大鼠将接受生理盐水、CPC-IL-1RA、未结合的 IL-1RA、mAv-CCR2RA 或未结合的 IA 注射 CCR2RA。将评估负重不对称和膝关节痛觉过敏；将评估 DRG 更改通过 IHC 和 RNAscope；将评估关节组织病理学，包括评估膝关节神经支配变化和免疫细胞浸润。这项具有竞争力的紧急修订将扩大已设计的影响阳离子疗法仅需一次 IA 给药即可持续抑制 OA 疼痛超过一个月除了疾病缓解（在原始 R01 中已解决）之外，还提供长期疼痛缓解。