Mahdavimehr Mohsen, Kaboudin Babak, Alaie Saied, Tondkar Farimah, Eshkaftaki Zahra Mahmoudi, Ebrahim-Habibi Mohammad-Bagher, Ghashghaee Mojtaba, Tahmasebi Elham, Zhang Tianjian, Gu Yanlong, Meratan Ali Akbar
Department of Biological Sciences, Institute for Advanced Studies in Basic Sciences (IASBS) Zanjan 45137-66731 Iran
Department of Chemistry, Institute for Advanced Studies in Basic Sciences (IASBS) Zanjan 45137-66731 Iran
RSC Adv. 2024 Oct 1;14(42):31227-31242. doi: 10.1039/d4ra02969k. eCollection 2024 Sep 24.
The main objective of the present study is to investigate the potency of new synthesized hydroxycarbamoyl phosphinic acid derivatives in modulating cytotoxic fibrillogenesis of hen egg white lysozyme (HEWL), as a common model in protein aggregation studies. Hydroxycarbamoyl phosphinic acid derivatives were prepared by the reaction of α-hydroxyalkylphosphinic acids with isocyanates (or isothiocyanates) in the presence of trimethylsilyl chloride (TMSCl). The designed process involves the condensation reaction leading to formation of new C sp-P bond formation. The synthesis and purity of novel designed compounds were confirmed by NMR, LC-MS, and HPLC techniques. A range of experiments, including thioflavin T (ThT) and 8-anilino-1-naphthalenesulfonic acid (ANS) fluorescence assays, Congo red binding measurement, atomic force microscopy imaging, MTT-based cell viability and hemolysis assays were employed to investigate anti-amyloidogenic effects of tested compounds. The obtained results demonstrate that these compounds are able to significantly modulate the self-assembly process of HEWL shortening of nucleation phase leading to the acceleration of fibrillation and appearance of very large and thick fibrils with decreased surface hydrophobicity and cytotoxicity. Based on ANS binding data, we suggest that increased exposure of hydrophobic patches of oligomeric species is the possible mechanism by which tested compounds promote self-assembly process of HEWL. Fluorescence anisotropy and molecular docking studies indicate the interaction of both synthesized compounds with HEWL, and more specifically with residues that are situated in the highly aggregation-prone β-domain region of protein. This study unveils the potential of hydroxyalkylphosphinic acids as modulators of amyloid fibrillation highlighting these compounds as a promising approach for targeting protein aggregates associated with neurodegenerative diseases.
本研究的主要目的是研究新合成的羟基氨基甲酰次膦酸衍生物调节鸡蛋清溶菌酶（HEWL）细胞毒性纤维形成的能力，HEWL是蛋白质聚集研究中的常见模型。羟基氨基甲酰次膦酸衍生物是通过α-羟基烷基次膦酸与异氰酸酯（或异硫氰酸酯）在三甲基氯硅烷（TMSCl）存在下反应制备的。设计的过程涉及缩合反应，导致形成新的C sp-P键。通过NMR、LC-MS和HPLC技术确认了新设计化合物的合成和纯度。采用了一系列实验，包括硫黄素T（ThT）和8-苯胺基-1-萘磺酸（ANS）荧光测定、刚果红结合测量、原子力显微镜成像、基于MTT的细胞活力和溶血测定，以研究测试化合物的抗淀粉样生成作用。获得的结果表明，这些化合物能够显著调节HEWL的自组装过程，缩短成核阶段，导致纤维形成加速，并出现非常大且厚的纤维，表面疏水性和细胞毒性降低。基于ANS结合数据，我们认为寡聚体物种疏水补丁暴露增加是测试化合物促进HEWL自组装过程的可能机制。荧光各向异性和分子对接研究表明，两种合成化合物均与HEWL相互作用，更具体地说是与位于蛋白质高度聚集倾向β结构域区域的残基相互作用。本研究揭示了羟基烷基次膦酸作为淀粉样纤维形成调节剂的潜力，突出了这些化合物作为靶向与神经退行性疾病相关的蛋白质聚集体的一种有前途的方法。
