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

项目摘要 尽管存在有希望的骨关节炎(OA)药物，但由于无效，其治疗仍然是一个挑战。 药物输送系统。当药物迅速从关节间隙和关节腔中清除时，关节内(IA)分娩是不够的 无法穿透带负电荷的致密软骨到达它们的细胞和基质靶点 最佳浓度的地点。因此，没有一种疾病修饰骨性关节炎药物(DMOADs)通过临床 由于担心全身毒性和缺乏软骨靶向而进行的试验。为了有效的治疗，至关重要的是 刺激疾病改变多个关节组织内的生物反应，包括软骨、滑膜和 软骨下骨。白细胞介素1受体拮抗剂(IL-1RA)被证明是一种有前途的DMOAD 在创伤后(PT)骨关节炎的临床前模型中调节滑膜炎症和软骨分解代谢； 然而，由于缺乏软骨靶向，关节停留时间短，未能显示出持续的临床效果。 时间到了。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。 用于提高阳离子药物或药物载体的摄取率、渗透深度和保留率。我们已经证明了 阳离子糖蛋白亲和素由于其最佳的大小和电荷，对于软骨内输送是有效的，因为 在大鼠和兔体内注射IA后，它迅速渗透到软骨的全层，导致400- 与中性软骨相比，软骨内摄取增加了一倍，并在软骨内保留了3-4 几周。基于亲和素的结构，我们设计了一种阳离子多肽载体(CPC)，其显示方式类似 在正常和糖胺多糖耗竭的软骨中都有很高的摄取。这个项目将向电气化方向发展。 通过将IL-1RA与亲和素和CPC偶联而带电，使其穿透并结合软骨，从而 增加其组织特异性和滞留时间。通过这种方式，软骨可以从屏障转化为药物 进入药库，使带电的IL-1RA在软骨和附近的抗分解代谢作用 与未修饰的IL-1RA相比，滑膜显著增强。在目标1中，亲和素-IL-1RA和CPC-IL-1RA 以及它们的主要传输特性(扩散率、平衡吸收、分配、结合 常数)与未修饰的IL-1RA在正常软骨和关节炎软骨中进行比较。目标2将评估 单次荷电IL-1RA抑制细胞因子诱导的软骨分解代谢的生物学效应 滑膜共培养骨性关节炎模型，比较抗生物素/CPC-IL-1RA结合物与单次和连续给药的比较 未修饰的IL-1RA。目的3将确定单次IA注射带电IL-1RA的治疗潜力 与未修饰的IL-1RA相比，使用兔PTOA模型。这项工作将推进基于电荷的药物领域的发展 应用基本概念，以多个关节组织为靶点进行有效的整体骨性关节炎治疗 生物静电学和生物传输学。这个基于收费的平台可用于交付广泛的 药物对其他性质相似的组织，如半月板、椎间盘和骨折痂，也 能够对由于缺乏组织靶向而未能通过临床试验的各种OA药物进行临床翻译。

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.