Institute of Biotechnology, University of Helsinki, Helsinki, 00014 (Finland).
ChemMedChem. 2012 Nov;7(11):1943-53. doi: 10.1002/cmdc.201200309. Epub 2012 Sep 11.
Acid α-glucosidase (GAA) is a lysosomal enzyme and a pharmacological target for Pompe disease, an inherited lysosomal storage disorder (LSD). An emerging treatment for LSDs is the use of pharmacological chaperones, small molecules that enhance total cellular activity of the target lysosomal protein. We have systematically studied thirteen inhibitors, which provide good lead compounds for the development of GAA chaperones. We have verified binding on GAA at low and neutral pH, mapping the range of pH during transport to lysosomes. These ligands inhibit GAA competitively and reversibly, and a few of the compounds show higher molecular stabilisation capacity than would be expected from their binding affinity. These molecules also increase lysosomal localisation of GAA variants in cells. In order to understand the specific molecular mechanism of the interactions, we docked the compounds to a homology model of the human GAA. Three factors contribute to the tightness of binding. Firstly, well-positioned hydroxy groups are essential to orient the ligand and make the binding specific. Secondly, the open nature of the GAA active site allows both large and small ligands to bind. The third and most important binding determinant is the positive charge on the ligand, which is neutralised by Asp 518 or Asp 616 on GAA. Our study creates a firm basis for the design of drugs to treat Pompe disease, as it provides a comparable study of the ligand properties. Our analysis suggests a useful drug design framework for specific pharmacological chaperones for human GAA.
酸性α-葡萄糖苷酶(GAA)是溶酶体酶,也是庞贝病(一种遗传性溶酶体贮积症)的药物靶点。新兴的溶酶体贮积症治疗方法是使用药理学伴侣,即增强靶溶酶体蛋白总细胞活性的小分子。我们已经系统地研究了十三种抑制剂,它们为 GAA 伴侣的开发提供了良好的先导化合物。我们已经验证了这些配体在低 pH 和中性 pH 下与 GAA 的结合,从而确定了溶酶体转运过程中的 pH 范围。这些配体竞争性且可逆性地抑制 GAA,其中一些化合物的分子稳定能力高于其结合亲和力所预期的水平。这些分子还增加了细胞中 GAA 变体的溶酶体定位。为了了解相互作用的具体分子机制,我们将这些化合物对接至人 GAA 的同源模型。有三个因素促成了结合的紧密性。首先,定位良好的羟基对于定向配体和使结合具有特异性至关重要。其次,GAA 活性位点的开放性允许大、小分子配体结合。第三个也是最重要的结合决定因素是配体上的正电荷,它被 GAA 上的 Asp518 或 Asp616 中和。我们的研究为治疗庞贝病的药物设计提供了坚实的基础,因为它提供了对配体性质的可比研究。我们的分析为人类 GAA 的特定药理学伴侣提出了一个有用的药物设计框架。