甲唑酰胺是一种碳酸酐酶（CA）抑制剂，具有令人满意的安全性。我们之前的研究已经证明甲唑酰胺在强直性脊柱炎（AS）中能提高CA1表达，并显示其治疗作用。本研究通过基因集富集分析（GSEA）和网络药理学研究，探索了CA1的致病作用以及甲唑酰胺在AS中的药理机制。十二个与CA1相关的基因集中有七个在AS组中富集。CA1在这七个基因集中是核心富集的，涉及到锌离子结合、芳香酯酶活性和一碳代谢过程。通过AS相关基因和甲唑酰胺靶基因的交集得到的候选靶基因进行功能分析表明，甲唑酰胺主要通过调节炎症途径，调控肿瘤坏死因子、IL-6和一氧化氮的产生，对AS产生治疗效应。我们选择了PTGS2、ESR1、GSK3β、JAK2、NOS2和CA1作为甲唑酰胺在AS中的治疗靶点。成功进行了分子对接和分子动力学模拟。此外，我们创新地获取了基因本体（GO）/京都基因和基因组百科全书（KEGG）分析结果与GSEA结果的交集，并发现甲唑酰胺在AS中的药理机制涉及骨矿化、血管生成、炎症和趋化因子信号通路等18个GO术语和5个KEGG术语。然而，需要通过体内/体外实验来验证这些机制。© 2023. Springer Nature Limited.

Methazolamide is a carbonic anhydrase (CA) inhibitor with satisfactory safety. Our previous studies have demonstrated the elevation of CA1 expression and the therapeutic effect of Methazolamide in Ankylosing spondylitis (AS). In this study, we explored the pathogenic role of CA1 and the pharmacological mechanism of Methazolamide in AS through Gene Set Enrichment Analysis (GSEA) and network pharmacology. Seven out of twelve CA1 related gene sets were enriched in AS group. CA1 was core enriched in above seven gene sets involving zinc ion binding, arylesterase activity and one carbon metabolic process. Functional analysis of the candidate target genes obtained from the intersection of AS associated genes and Methazolamide target genes indicated that Methazolamide exerts therapeutic effects on AS mainly through inflammatory pathways which regulate the production of tumor necrosis factor, IL-6 and nitric oxide. PTGS2, ESR1, GSK3β, JAK2, NOS2 and CA1 were selected as therapeutic targets of Methazolamide in AS. Molecular docking and molecular dynamics simulations were performed successfully. In addition, we innovatively obtained the intersection of Gene Ontology (GO)/Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses and GSEA results, and found that 18 GO terms and 5 KEGG terms were indicated in the pharmacological mechanism of Methazolamide in AS, involving bone mineralization, angiogenesis, inflammation, and chemokine signaling pathways. Nevertheless, validation for these mechanisms is needed in vivo/vitro experiments.© 2023. Springer Nature Limited.