Department of Pharmacology, Faculty of Pharmacy, Ziauddin University, Karachi, Pakistan.
Department of Molecular Medicine, Ziauddin University, Karachi, Pakistan.
PLoS One. 2024 Jul 25;19(7):e0307708. doi: 10.1371/journal.pone.0307708. eCollection 2024.
Advanced glycation end products (AGEs) play an important role in the pathogenesis of age-linked disorders and diabetes mellitus. The aim of this study was to assess the repurposing potential of Phloroglucinol (PHL the antispasmodic drug), as an anti-glycation agent using Fructose-BSA model. The ability of PHL to inhibit AGE formation was evaluated using AGEs formation (Intrinsic fluorescence), fructosamine adduct (NBT) and free lysine availability (TNBSA) assays. The BSA protein conformation was assessed through Thioflavin-T, Congo-Red and Circular Dichroism assays. The lysine blockade and carbonyl entrapment were explored as possible mode of action. Our data showed that PHL significantly decreased the formation of AGEs with an IC50 value of 0.3mM. The fructosamine adducts and free lysine load was found to be reduced. Additionally, the BSA conformation was preserved by PHL. Mechanistic assays did not reveal involvement of lysine blockade as underlying reason for reduction in AGEs load. This was also supported by computational data whereby PHL failed to engage any catalytic residue involved in early fructose-BSA interaction. However, it was found to entrap the carbonyl moieties. In conclusion, the PHL demonstrated anti-glycation potential, which can be attributed to its ability to entrap carbonyl intermediates. Hence, the clinically available antispasmodic drug, presents itself as a promising candidate to be repurposed as anti-glycation agent.
糖基化终产物（AGEs）在与年龄相关的疾病和糖尿病的发病机制中起着重要作用。本研究旨在评估间苯三酚（PHL，一种抗痉挛药物）作为抗糖基化剂在果糖-BSA 模型中的再利用潜力。通过 AGEs 形成（固有荧光）、果糖胺加合物（NBT）和游离赖氨酸可用性（TNBSA）测定来评估 PHL 抑制 AGE 形成的能力。通过硫代黄素-T、刚果红和圆二色性测定来评估 BSA 蛋白构象。探讨了赖氨酸封锁和羰基捕获作为可能的作用模式。我们的数据表明，PHL 可显著降低 AGEs 的形成，IC50 值为 0.3mM。发现果糖胺加合物和游离赖氨酸负荷减少。此外，PHL 还能保持 BSA 的构象。机制研究表明，赖氨酸封锁不是降低 AGEs 负荷的原因。这也得到了计算数据的支持，因为 PHL 未能与早期果糖-BSA 相互作用中涉及的任何催化残基结合。然而，它被发现可以捕获羰基部分。总之，PHL 表现出抗糖基化的潜力，这可归因于其捕获羰基中间产物的能力。因此，临床上可用的抗痉挛药物本身就是一种很有前途的抗糖基化剂再利用候选药物。
