Villano Anthea, Barcaro Giovanni, Monti Susanna, Barbani Niccoletta, Rizzo Antonio, Rossin Daniela, Rastaldo Raffaella, Giachino Claudia, Cristallini Caterina
CNR-IPCF, National Research Council-Institute for Chemical and Physical Processes, Area della Ricerca, Via Moruzzi 1, I-56124 Pisa, Italy.
CNR-ICCOM, National Research Council-Institute of Chemistry of Organometallic Compounds, Area della Ricerca, Via Moruzzi 1, I-56124 Pisa, Italy.
Biomedicines. 2022 Aug 24;10(9):2070. doi: 10.3390/biomedicines10092070.
The recent advances in nanotechnology are revolutionizing preventive and therapeutic approaches to treating cardiovascular diseases. Controlling the extracellular matrix metalloproteinase (MMP) activation and expression in the failing human left ventricular myocardium represents a significant therapeutic target for heart disease. In this study, we used molecularly imprinting polymers (MIPs) to restore the correct balance between MMPs and their tissue inhibitors (TIMPs), and explored the potential of this technique exhaustively through chemical synthesis, physicochemical and biological characterizations, and computational chemistry methods. By molecular dynamics simulations based on classical force fields, we simulated the early stages of the imprinting process in solution disclosing the pivotal interaction established between the monomers and the MMP9 protein template. The average interaction energies of methacrylic acid (MAA) and poly (ethylene glycol) ethyl ether methacrylate (PEG) units were in the ranges 17-22 and 30-37 kcal/mol, respectively. At low coverage, the PEG monomers seemed firmly anchored to the protein surface and were not displaced by water, while only about 20% of MAA was replaced by water. The synthesis of MIPs was successfully with a monomer conversion higher than 99% and the production of spherical particles with average diameter of 344 ± 33 nm. HPLC analysis showed a specific recognition factor of MMP9 on MIPs of about 1.3. FT-IR Chemical Imaging confirmed the mechanisms necessary to generate a "selective memory" of the MIPs towards the enzyme. HPLC results indicated that the rebound amount of both TIMP1 and MMP2 to MIPs is lower than that of the template, showing a selectivity factor of 2.1 and 2.3, respectively. Preliminary tests on the effect of MIPs on H9C2 cells revealed that this treatment has no cytotoxic effects.
纳米技术的最新进展正在彻底改变心血管疾病的预防和治疗方法。控制衰竭的人类左心室心肌细胞外基质金属蛋白酶(MMP)的激活和表达是心脏病的一个重要治疗靶点。在本研究中,我们使用分子印迹聚合物(MIP)来恢复MMP与其组织抑制剂(TIMP)之间的正确平衡,并通过化学合成、物理化学和生物学表征以及计算化学方法全面探索了该技术的潜力。通过基于经典力场的分子动力学模拟,我们模拟了溶液中印迹过程的早期阶段,揭示了单体与MMP9蛋白模板之间建立的关键相互作用。甲基丙烯酸(MAA)和聚(乙二醇)甲基丙烯酸乙酯(PEG)单元的平均相互作用能分别在17 - 22和30 - 37 kcal/mol范围内。在低覆盖率下,PEG单体似乎牢固地锚定在蛋白质表面,不会被水取代,而只有约20%的MAA被水取代。成功合成了MIP,单体转化率高于99%,并制备出平均直径为344±33 nm的球形颗粒。HPLC分析表明,MIP对MMP9的特异性识别因子约为1.3。傅里叶变换红外光谱化学成像证实了MIP对该酶产生“选择性记忆”所需的机制。HPLC结果表明,TIMP1和MMP2与MIP的回跳量均低于模板,选择性因子分别为2.1和2.3。对MIP对H9C2细胞影响的初步测试表明,这种处理没有细胞毒性作用。