Breland Austen N, Ross Matthew K, Fitzkee Nicholas C, Elder Steven H
Department of Agricultural & Biological Engineering, Mississippi State University, Starkville, MS 39762, USA.
Department of Comparative Biomedical Sciences, Mississippi State University, Starkville, MS 39762, USA.
Int J Mol Sci. 2025 Apr 25;26(9):4093. doi: 10.3390/ijms26094093.
ADAMTS-5 (aggrecanase-2) is a major metalloprotease involved in regulating the cartilage extracellular matrix. Due to its role in removing aggrecan in the progression of osteoarthritis (OA), ADAMTS-5 is often regarded as a potential therapeutic target for OA. Punicalagin (PCG), a polyphenolic ellagitannin found in pomegranate ( L.), and ellagic acid (EA), a hydrolytic metabolite of PCG, have been widely investigated as potential disease-modifying osteoarthritis drugs (DMOADs) due to their potent antioxidant and anti-inflammatory properties, but their interaction with ADAMTS-5 has yet to be determined. In this study, molecular docking simulations were used to predict enzyme-inhibitor binding interactions. The results suggest that both compounds may be able to bind within the active site via the formation of H bonds and interactions between the ligand's aromatic rings and hydrophobic residue in the enzyme with inhibition constants of 183.3 µM and 1.13 µM for PCG and EA, respectively. Biochemical activity against recombinant human ADAMTS-5 was assessed using a dimethylmethylene blue-based assay to determine residual sulfated glycosaminoglycan (sGAG) in porcine articular cartilage. Although its loss could not be attributed to ADAMTS-5, sGAG was effectively persevered by PCG and EA. The potential conversion of PCG to EA by enzyme-catalyzed hydrolysis activity was then investigated using liquid chromatography-mass spectroscopy to determine the potential for the use of PCG and EA as a prodrug-proactive metabolite pair in the development of drug delivery systems to arthritic synovial joints.
ADAMTS-5（软骨聚集蛋白聚糖酶-2）是一种参与调节软骨细胞外基质的主要金属蛋白酶。由于其在骨关节炎（OA）进展过程中去除聚集蛋白聚糖的作用，ADAMTS-5常被视为OA的潜在治疗靶点。石榴皮素（PCG）是石榴中发现的一种多酚类鞣花单宁，而鞣花酸（EA）是PCG的水解代谢产物，由于其强大的抗氧化和抗炎特性，已被广泛研究作为潜在的改善病情的骨关节炎药物（DMOADs），但它们与ADAMTS-5的相互作用尚未确定。在本研究中，使用分子对接模拟来预测酶-抑制剂结合相互作用。结果表明，这两种化合物都可能通过形成氢键以及配体芳香环与酶中疏水残基之间的相互作用而结合在活性位点内，PCG和EA的抑制常数分别为183.3 μM和1.13 μM。使用基于二甲基亚甲基蓝的测定法评估对重组人ADAMTS-5的生化活性，以确定猪关节软骨中残留的硫酸化糖胺聚糖（sGAG）。尽管sGAG的损失不能归因于ADAMTS-5，但PCG和EA有效地保留了sGAG。然后使用液相色谱-质谱法研究酶催化水解活性将PCG转化为EA的可能性，以确定在向关节炎滑膜关节递送药物的系统开发中使用PCG和EA作为前药-前体代谢物对的可能性。
