Harnessing polyhydroxylated pyrrolidines as a stabilizer of acid alpha-glucosidase (GAA) to enhance the efficacy of enzyme replacement therapy in Pompe disease

• Published in: Bioorganic & medicinal chemistry Vol. 78; p. 117129
• Main Authors: Li, Huang-Yi, Lee, Ni-Chung, Chiu, Yu-Ting, Chang, Yu-Wen, Lin, Chu-Chung, Chou, Cheng-Li, Chien, Yin-Hsiu, Hwu, Wuh-Liang, Cheng, Wei-Chieh
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 15.01.2023
• Subjects: Acid alpha-glucosidase (GAA); alpha-Glucosidases; Chemical space; Combinatorial chemistry; Cyclic nitrone; Enzyme Replacement Therapy; Glycogen Storage Disease Type II - diagnosis; Glycogen Storage Disease Type II - drug therapy; Humans; Iminosugar; Polyhydroxylated pyrrolidine; Pompe disease; Protein stabilizer; Protein thermal shift assay; Pompe disease; Cyclic nitrone; Iminosugar; Chemical space; Protein stabilizer; Polyhydroxylated pyrrolidine; Combinatorial chemistry; Acid alpha-glucosidase (GAA); Protein thermal shift assay
• ISSN: 0968-0896; 1464-3391
• DOI: 10.1016/j.bmc.2022.117129

[Display omitted] •A 16-membered ADMDP library including all diastereomers was evaluated.•The best configuration pattern of pyrrolidines was identified to stabilize GAA.•The derivatives were synthesized through internal and external modifications.•The most potent stabilizer 21 was found to prevent rh-GAA from denaturation.•Stabilizer 21 showed a 5.1-fold increase in rh-GAA activity in Pompe fibroblasts. To discover small molecules as acid alpha-glucosidase (GAA) stabilizers for potential benefits of the exogenous enzyme treatment toward Pompe disease cells, we started from the initial screening of the unique chemical space, consisting of sixteen stereoisomers of 2-aminomethyl polyhydroxylated pyrrolidines (ADMDPs) to find out two primary stabilizers 17 and 18. Further external or internal structural modifications of 17 and 18 were performed to increase structural diversity, followed by the protein thermal shift study to evaluate the GAA stabilizing ability. Fortunately, pyrrolidine 21, possessing an l-arabino-typed configuration pattern, was identified as a specific potent rh-GAA stabilizer, enabling the suppression of rh-GAA protein denaturation. In a cell-based Pompe model, co-administration of 21 with rh-GAA protein significantly improved enzymatic activity (up to 5-fold) compared to administration of enzyme alone. Potentially, pyrrolidine 21 enables the direct increase of ERT (enzyme replacement therapy) efficacy in cellulo and in vivo.