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

• 批准号: 9887607
• 负责人: Ambika Goel Bajpayee
• 金额: $ 46.12万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-21 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9887607
• 关键词: Adverse effects; Affect; Anabolism; Arthritis; Avidin; Binding; Biochemical; Biodistribution; Biological; Biomechanics; Bone callus; Bovine Cartilage; Caliber; Cartilage; Cartilage Matrix; Catabolism; Cations; Cell Survival; Cells; Charge; Clinical Trials; Coculture Techniques; Collagen; Confocal Microscopy; Degenerative polyarthritis; Diffuse; Disease; Dose; Drug Carriers; Drug Delivery Systems; Drug or chemical Tissue Distribution; Effectiveness; Electrostatics; Engineering; Equilibrium; Fracture; Gene Expression; Gene Proteins; Glycoproteins; Glycosaminoglycans; Histology; Immunohistochemistry; In Vitro; Inflammation; Inflammatory; Injury; Interleukin-1; Interleukin-1 Receptors; Interleukins; Intervertebral disc structure; Intra-Articular Injections; Joints; Lead; Mass Spectrum Analysis; Measures; Mediating; Meniscus structure of joint; MicroRNAs; Modeling; Modification; Organ; Oryctolagus cuniculus; Outcome; Penetration; Peptides; Pharmaceutical Preparations; Pre-Clinical Model; Property; Proteins; Proteomics; Radiolabeled; Rattus; Role; Site; Specificity; Structure; Synovial Fluid; Synovial Membrane; Synovial joint; Techniques; Testing; Therapeutic; Thick; Time; Tissues; Western Blotting; Work; affinity labeling; anakinra; articular cartilage; base; bone; clinical effect; clinical translation; cytokine; density; design; driving force; drug clearance; effective therapy; extracellular; in vivo; inhibitor/antagonist; joint injury; microCT; prevent; protein expression; residence; response; subchondral bone; systemic toxicity; targeted delivery; time use; tissue degeneration; uptake

Project Summary Despite the existence of promising osteoarthritis (OA) drugs, its treatment remains a challenge due to ineffective drug delivery systems. Intra-articular (IA) delivery is inadequate as drugs rapidly clear out from joint space and are unable to penetrate through the dense, negatively charged cartilage and reach their cell and matrix target sites at optimal concentrations. As a result, no disease modifying OA drugs (DMOADs) have passed clinical trials due to concerns of systemic toxicity and lack of cartilage targeting. For effective treatment, it is critical to stimulate a disease modifying biological response within multiple joint tissues, including cartilage, synovium and subchondral bone. Interleukin (IL)-1 receptor antagonist (IL-1RA) is proven to be a promising DMOAD for modulating both synovium inflammation and cartilage catabolism in preclinical models of post-traumatic (PT)OA; however, it has failed to show sustained clinical effect owing to lack of cartilage targeting and short joint residence time. The high negative charge density of cartilage provides a unique opportunity to use electrostatic interactions for enhancing uptake, depth of penetration, and retention of cationic drugs or drug carriers. We have shown that the cationic glycoprotein Avidin, owing to its optimal size and charge, was effective for intra-cartilage delivery as it rapidly penetrated through full thickness of cartilage in rats and rabbits following IA injection, resulting in 400- fold higher intra-cartilage uptake compared to its neutral counterpart and was retained inside cartilage for 3-4 weeks. Based on Avidin’s structure, we have designed a Cationic Peptide Carrier (CPC) that displayed similarly high uptake in both normal and glycosaminoglycan-depleted cartilage. This project will develop electrically charged IL-1RA by conjugating it with Avidin and CPC to make it cartilage penetrating and binding, thus increasing its tissue specificity and residence time. This way, cartilage can be converted from a barrier to drug entry into a drug depot, such that the anti-catabolic effects of charged IL-1RA in both cartilage and nearby synovium are significantly enhanced compared to unmodified IL-1RA. In Aim 1, Avidin-IL-1RA and CPC-IL-1RA will be characterized and their key transport properties (diffusivities, equilibrium uptakes, partitioning, binding constants) will be compared with unmodified IL-1RA in normal and arthritic cartilage. Aim 2 will evaluate the biological efficacy of a single dose of charged IL-1RA for inhibiting cytokine induced catabolism in a cartilage- synovium co-culture OA model, comparing Avidin/CPC-IL-1RA conjugates with single and continuous dose of unmodified IL-1RA. Aim 3 will determine the therapeutic potential of a single IA injection of charged IL-1RA relative to unmodified IL-1RA using a rabbit PTOA model. This work will advance the field of charge based drug delivery in targeting multiple joint tissues for effective, holistic OA treatment by applying fundamental concepts of bio-electrostatics and bio-transport. This charge-based platform can be used for delivering a wide range of drugs to other tissues with similar properties, such as meniscus, intervertebral disc and fracture callus, and also enable clinical translation of various OA drugs that have failed clinical trials due to lack of tissue targeting.

项目摘要 尽管存在有前途的骨关节炎（OA）药物，但由于无效，其治疗仍然是一个挑战。 药物递送系统。关节内（IA）给药不足，因为药物会迅速从关节间隙清除， 无法穿透致密的带负电荷的软骨并到达它们的细胞和基质目标 在最佳浓度的位置。因此，没有疾病修饰OA药物（DMOAD）通过临床试验。 由于担心全身毒性和缺乏软骨靶向，为了有效治疗， 刺激多种关节组织内的疾病修饰生物反应，包括软骨、滑膜和 软骨下骨白细胞介素-1受体拮抗剂（IL-1 RA）被证明是一种很有前途的DMOAD， 在创伤后（PT）OA的临床前模型中调节滑膜炎症和软骨炎性反应; 然而，由于缺乏软骨靶向和关节驻留时间短， 时间软骨的高负电荷密度为利用静电相互作用提供了独特的机会 用于增强阳离子药物或药物载体的吸收、渗透深度和保留。我们已经证明 阳离子糖蛋白抗生物素蛋白由于其最佳的尺寸和电荷，对于软骨内递送是有效的， 在IA注射后，它迅速穿透大鼠和兔的软骨全层，导致400- 与中性对应物相比，软骨内吸收高出1倍，并保留在软骨内3-4 周基于亲和素的结构，我们设计了一种阳离子肽载体（CPC）， 在正常和糖胺聚糖耗尽的软骨中的高摄取。该项目将开发电动 将IL-1 RA与抗生物素蛋白和CPC偶联，使其具有软骨穿透性和结合性， 增加其组织特异性和停留时间。这样，软骨就可以从一个屏障转化为药物， 进入药物贮库，使得带电荷的IL-1 RA在软骨和附近的抗分解代谢作用 与未修饰的IL-1 RA相比，滑膜中的IL-1 RA显著增强。在目标1中，亲和素-IL-1 RA和CPC-IL-1 RA 将被表征，它们的关键传输特性（扩散系数、平衡吸收、分配、结合 常数）与正常和关节炎软骨中未修饰的IL-1 RA进行比较。目标2将评估 单一剂量的荷电IL-1 RA抑制软骨中细胞因子诱导的软骨炎的生物学功效， 滑膜共培养OA模型，比较抗生物素蛋白/CPC-IL-1 RA缀合物与单次和连续剂量的 未修饰的IL-1 RA。目的3将确定单次IA注射荷电IL-1 RA的治疗潜力 相对于未修饰的IL-1 RA，使用兔PTOA模型。这项工作将推动电荷药物领域的发展 通过应用基本概念，靶向多个关节组织进行有效的整体OA治疗 生物静电学和生物运输学。这种基于收费的平台可用于提供各种 药物对具有类似性质的其他组织，如半月板、椎间盘和骨折骨痂， 使临床翻译的各种OA药物，已失败的临床试验，由于缺乏组织靶向。