Department of Bioengineering, Northeastern University, Boston, MA, USA.
Department of Biochemistry, Northeastern University, Boston, MA, USA.
Osteoarthritis Cartilage. 2023 Jun;31(6):780-792. doi: 10.1016/j.joca.2023.01.573. Epub 2023 Feb 3.
Blocking the interleukin-1 (IL-1) catabolic cascade following joint trauma can be achieved using its receptor antagonist, IL-1Ra. However, its clinical translation for osteoarthritis therapy has been unsuccessful due to its rapid joint clearance and lack of targeting and penetration into deep cartilage layers at therapeutic concentrations. Here, we target the high negative charge of cartilage aggrecan-glycosaminoglycans (GAGs) by attaching cationic carriers to IL-1Ra. IL-1Ra was conjugated to the cartilage targeting glycoprotein, Avidin, and a short length optimally charged cationic peptide carrier (CPC+14). It is hypothesized that electro-diffusive transport and binding properties of IL-1Ra-Avidin and IL-1Ra-CPC+14 will create intra-cartilage depots of IL-1Ra, resulting in long-term suppression of IL-1 catabolism with only a single administration.
IL-1Ra was conjugated to Avidin or CPC+14 using site specific maleimide linkers, and confirmed using gel electrophoresis, high-performance liquid chromatography (HPLC), and mass spectrometry. Intra-cartilage transport and retention of conjugates was compared with native IL-1Ra. Attenuation of IL-1 catabolic signaling with one-time dose of IL-1Ra-CPC+14 and IL-1Ra-Avidin was assessed over 16 days using IL-1α challenged bovine cartilage and compared with unmodified IL-1Ra.
Positively charged IL-1Ra penetrated through the full-thickness of cartilage, creating a drug depot. A single dose of unmodified IL-1Ra was not sufficient to attenuate IL-1-induced cartilage deterioration over 16 days. However, when delivered using Avidin, and to a greater extent CPC+14, IL-1Ra significantly suppressed cytokine induced GAG loss and nitrite release while improving cell metabolism and viability.
Charge-based cartilage targeting drug delivery systems hold promise as they can enable long-term therapeutic benefit with only a single dose.
通过使用白细胞介素-1（IL-1）受体拮抗剂 IL-1Ra 阻断关节创伤后的 IL-1 分解级联反应。然而，由于其在治疗浓度下在关节中快速清除以及缺乏靶向性和穿透到深层软骨层，其在骨关节炎治疗中的临床转化尚未成功。在这里，我们通过将阳离子载体附着到 IL-1Ra 上来靶向软骨聚集蛋白聚糖（GAGs）的高负电荷。IL-1Ra 与软骨靶向糖蛋白亲和素偶联，并与短长度的最佳带电阳离子肽载体（CPC+14）偶联。假设 IL-1Ra-亲和素和 IL-1Ra-CPC+14 的电扩散转运和结合特性将在软骨内创建 IL-1Ra 的储库，从而仅通过单次给药即可长期抑制 IL-1 的分解代谢。
使用位点特异性马来酰亚胺接头将 IL-1Ra 偶联到亲和素或 CPC+14 上，并通过凝胶电泳、高效液相色谱（HPLC）和质谱进行确认。将偶联物的软骨内转运和保留与天然 IL-1Ra 进行比较。使用单次剂量的 IL-1Ra-CPC+14 和 IL-1Ra-亲和素评估 16 天内 IL-1α 挑战的牛软骨中 IL-1 分解代谢信号的衰减，并与未修饰的 IL-1Ra 进行比较。
带正电荷的 IL-1Ra 穿透软骨的全厚度，形成药物储库。单次给予未修饰的 IL-1Ra 不足以在 16 天内减弱 IL-1 诱导的软骨退化。然而，当使用亲和素，并且在更大程度上使用 CPC+14 时，IL-1Ra 显著抑制细胞因子诱导的 GAG 丢失和亚硝酸盐释放，同时改善细胞代谢和活力。
基于电荷的软骨靶向药物递送系统具有很大的应用前景，因为它们可以通过单次给药实现长期的治疗效果。
