BackgroundMycobacterium tuberculosis (Mtb) carbonic anhydrases (CAs) are critical enzymes that regulate pH by converting CO2 to HCO3−, essential for Mtb’s survival in acidic environments. Inhibiting γ-CAs presents a potential target for novel antituberculosis drugs with unique mechanisms of action.ObjectiveThis study aimed to explore the biological connections underlying Mtb pathogenesis and investigate the mechanistic actions of antituberculosis compounds targeting the Cas9 protein.MethodsWe employed homology modeling and virtual screening to identify compounds with high binding affinities for Cas9 protein. This study used the homology modeling approach employing high-quality AlphaFold DB models for γ-CA. Furthermore, the systems biology approach was used for analyzing the integrated modelling of compounds, integrating data on genes, pathways, phenotypes, and molecular descriptors. Single-cell RNA sequencing was also conducted to profile gene expression.ResultsThree compounds, F10921405, F08060425, and F14437079, potentially binding to Cas9 protein, have been identified. F10921405 and F08060425 showed significant overlap in their effects on pathways related to the immune response, while F14437079 displayed distinct mechanistic pathways. Expression profiling revealed high levels of genes such as PDE4D, ROCK2, ITK, MAPK10, and SYK in response to F1092–1405 and F0806-0425, and MMP2 and CALCRL in response to F1443-7079. These genes, which play a role in immune modulation and lung tissue integrity, are essential to fight against Mtb.ConclusionThe molecular relationship and pathways linked to the mentioned compounds give the study a holistic perspective of targeting Mtb, which is essential in designing specific therapeutic approaches. Subsequent research will involve experimental validation to demonstrate the efficacy of the promising candidates in Mtb infections.

中文翻译：

---

化学信息学和网络药理学方法阐明结核分枝杆菌γ-碳酸酐酶抑制剂的作用机制


背景结核分枝杆菌 (Mtb) 碳酸酐酶 (CA) 是通过将 CO2 转化为 HCO3− 来调节 pH 值的关键酶，这对于 Mtb 在酸性环境中的生存至关重要。抑制γ-CAs为具有独特作用机制的新型抗结核药物提供了潜在靶点。 目的本研究旨在探讨Mtb发病机制的生物学联系，并研究针对Cas9蛋白的抗结核化合物的作用机制。鉴定与 Cas9 蛋白具有高结合亲和力的化合物。本研究使用同源建模方法，采用高质量的 AlphaFold DB 模型来构建 γ-CA。此外，系统生物学方法用于分析化合物的综合建模，整合基因、途径、表型和分子描述符的数据。还进行了单细胞 RNA 测序来分析基因表达。结果已鉴定出可能与 Cas9 蛋白结合的三种化合物：F10921405、F08060425 和 F14437079。 F10921405和F08060425对免疫反应相关途径的影响显示出显着的重叠，而F14437079则显示出不同的机制途径。表达谱显示高水平的基因，例如响应 F1092-1405 和 F0806-0425 的 PDE4D、ROCK2、ITK、MAPK10 和 SYK，以及响应 F1443-7079 的 MMP2 和 CALCRL。这些基因在免疫调节和肺组织完整性中发挥作用，对于对抗 Mtb 至关重要。结论与上述化合物相关的分子关系和途径为研究提供了靶向 Mtb 的整体视角，这对于设计特定的治疗方法至关重要。 后续研究将涉及实验验证，以证明有希望的候选药物在结核分枝杆菌感染中的功效。