Dipartimento di Chimica e Chimica Industriale, Via Giuseppe Moruzzi, 13, 56124 Pisa, Italy.
Department of Biothermodynamics and Drug Design, Institute of Biotechnology, Life Sciences Center, Vilnius University, Saulėtekio 7, Vilnius LT-10257, Lithuania.
Bioorg Chem. 2021 Oct;115:105155. doi: 10.1016/j.bioorg.2021.105155. Epub 2021 Jul 9.
Matrix metalloproteinases (MMPs) are an important factor in cancer progression and metastasis, especially gelatinases MMP-2 and MMP-9. A simple methodology for their detection and monitoring is highly desirable. Molecular probes have been very widely and successfully applied to study the activity of MMPs in cellular processes in vitro. We thus synthesized a small compound library of MMP-2 and MMP-9 binding probes based on drug molecules and endowed with free amine groups for the functionalization of transducer surfaces. In this study, we combined experimental results obtained by a kinetic fluorogenic peptide substrate cleavage assay with molecular modeling studies in order to assess the ability of the probe to bind to their target enzymes. The synthesized biphenyl substituted lysine derivatives showed IC-values in the low nanomolar concentration range against MMP-2 (ligands 3a-d: 3 nM to 8 µM, ligands 4a-d: 45 nM to 350 µM) and low micromolar range against MMP-9 (ligands 3a-d: 350 nM to 60 µM, ligands 4a-d: 5 µM to 600 µM), with a selectivity up to more than 160-fold for MMP-2. The experimental results correlated well with molecular modelling with FleXAID and X-score functions. We showed that in our compound series, the side chain remained far away from the S1' cavity and the ligand for all the docked minima. Ligands 4a-d with their free amine group on the side chain may thus be bound to transducer surfaces for the fabrication of sensors, while retaining their activity against their target enzymes.
基质金属蛋白酶（MMPs）是癌症进展和转移的重要因素，尤其是明胶酶 MMP-2 和 MMP-9。非常需要一种简单的方法来检测和监测它们。分子探针已被广泛而成功地应用于研究 MMPs 在细胞过程中的活性。因此，我们基于药物分子合成了 MMP-2 和 MMP-9 结合探针的小分子化合物库，并赋予了用于传感器表面功能化的游离胺基。在这项研究中，我们结合了通过动力学荧光肽底物切割测定获得的实验结果和分子建模研究，以评估探针与靶酶结合的能力。合成的联苯取代赖氨酸衍生物对 MMP-2 的 IC 值在纳摩尔低浓度范围内（配体 3a-d：3 nM 至 8 µM，配体 4a-d：45 nM 至 350 µM），对 MMP-9 的 IC 值在微摩尔低浓度范围内（配体 3a-d：350 nM 至 60 µM，配体 4a-d：5 µM 至 600 µM），对 MMP-2 的选择性高达 160 倍以上。实验结果与 FleXAID 和 X-score 函数的分子建模很好地相关。我们表明，在我们的化合物系列中，侧链远离 S1'腔，配体对于所有对接的最小值都是如此。带有游离胺基的侧链的配体 4a-d 因此可以与传感器表面结合以制造传感器，同时保留其对靶酶的活性。
