El Gammal Reem N, Elmansi Heba, El-Emam Ali A, Belal Fathalla, Elzahhar Perihan A, Belal Ahmed S F, Hammouda Mohammed E A
Department of Medicinal Chemistry, Faculty of Pharmacy, Mansoura University, Mansoura, 35516, Egypt.
Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Mansoura University, Mansoura, 35516, Egypt.
BMC Chem. 2023 Apr 6;17(1):31. doi: 10.1186/s13065-023-00944-z.
In this work, the binding mechanism between donepezil (DNP) and bovine serum albumin (BSA) was established using several techniques, including fluorimetry, UV- spectrophotometry, synchronous fluorimetry (SF), fourier transform infrared (FTIR), fluorescence resonance energy transfer (FRET) besides molecular docking study. The fluorescence quenching mechanism of DNP-BSA binding was a combined dynamic and static quenching. The thermodynamic parameters, binding forces, binding constant, and the number of binding sites were determined using a different range of temperature settings. Van't Hoff's equation was used to calculate the reaction parameters, including enthalpy change (ΔH) and entropy change (ΔS). The results pointed out that the DNP-BSA binding was endothermic. It was shown that the stability of the drug-protein system was predominantly due to the intermolecular hydrophobic forces. Additionally, the site probing method revealed that subdomain IIA (Site I) is where DNP and BSA's binding occurs. This was validated using a molecular docking study with the most stable DNP configuration. This study might help to understand DNP's pharmacokinetics profile and toxicity as well as provides crucial information for its safe use and avoiding its toxicity.
在本研究中，采用多种技术建立了多奈哌齐（DNP）与牛血清白蛋白（BSA）之间的结合机制，这些技术包括荧光法、紫外分光光度法、同步荧光法（SF）、傅里叶变换红外光谱法（FTIR）、荧光共振能量转移法（FRET）以及分子对接研究。DNP与BSA结合的荧光猝灭机制是动态猝灭和静态猝灭的结合。利用不同的温度设置范围测定了热力学参数、结合力、结合常数和结合位点数。使用范特霍夫方程计算反应参数，包括焓变（ΔH）和熵变（ΔS）。结果表明，DNP与BSA的结合是吸热的。结果表明，药物 - 蛋白质体系的稳定性主要归因于分子间的疏水作用力。此外，位点探测方法表明亚结构域IIA（位点I）是DNP与BSA发生结合的位置。这通过对最稳定的DNP构型进行分子对接研究得到了验证。本研究可能有助于了解DNP的药代动力学特征和毒性，并为其安全使用和避免毒性提供关键信息。
