Dept. of Chemistry, University of British Columbia, Vancouver, BC, V6T 1Z1, Canada.
Dept. of Biochemistry and Microbiology, University of Victoria, Victoria, BC, V8W 3P6, Canada.
Angew Chem Int Ed Engl. 2022 Sep 19;61(38):e202207974. doi: 10.1002/anie.202207974. Epub 2022 Aug 12.
Gaucher disease is a lysosomal storage disorder caused by mutations which destabilize the native folded form of GCase, triggering degradation and ultimately resulting in low enzyme activity. Pharmacological chaperones (PCs) which stabilize mutant GCase have been used to increase lysosomal activity through improving trafficking efficiency. By engineering their inherent basicity, we have synthesized PCs that change conformation between the ER and the lysosomal environment, thus weakening binding to GCase after its successful trafficking to the lysosome. NMR studies confirmed the conformational change while X-ray data reveal bound conformations and binding modes. These results were further corroborated by cell studies showing increases in GCase activity when using the pH-switchable probe at low dosing. Preliminary in vivo assays with humanized mouse models of Gaucher showed enhanced GCase activity levels in relevant tissues, including the brain, further supporting their potential.
戈谢病是一种溶酶体贮积症，由突变引起，这些突变会破坏 GCase 的天然折叠形式，引发降解，最终导致酶活性降低。已使用稳定突变 GCase 的药理学伴侣（PC）通过提高运输效率来增加溶酶体活性。通过对其固有碱性的工程改造，我们合成了 PC，它们在 ER 和溶酶体环境之间改变构象，从而在 GCase 成功运输到溶酶体后减弱与 GCase 的结合。NMR 研究证实了构象变化，而 X 射线数据则揭示了结合构象和结合模式。细胞研究进一步证实了这一结果，表明使用 pH 可切换探针进行低剂量给药时，GCase 活性增加。使用戈谢病人源化小鼠模型进行的初步体内试验显示，相关组织（包括大脑）中的 GCase 活性水平提高，进一步支持了它们的潜力。
