在本研究中，我们研究了茶碱与不同天然产物化合物的结合，希望构建新的杂交物，同时对胆碱能和炎症途径具有双重活性，作为治疗阿尔茨海默病（AD）的潜在药物。在28种经过测试的杂交物中，两种杂交物，acefylline-eugenol 6d和acefylline-isatin 19，能够以低微摩尔浓度抑制乙酰胆碱酯酶（AChE），其IC50值分别为1.8和3.3μM，与加兰他敏标准AChE抑制剂相比。此外，所制备的杂交物对脂多糖诱导的RAW 264.7细胞炎症具有显著的抗炎作用，并以剂量依赖的方式降低了一氧化氮（NO）、肿瘤坏死因子α（TNF-α）、白细胞介素-1β（IL-1β）和白细胞介素-6（IL-6）的水平。这些杂交物对RAW 264.7细胞中由脂多糖（LPS）诱导的一氧化氮（NO）、肿瘤坏死因子α（TNF-α）、白细胞介素-1β（IL-1β）和白细胞介素-6（IL-6）水平的显著降低。本研究的结果进一步通过网络药理学分析进行了解释，提示乙酰胆碱酯酶和一氧化氮合酶是最活性化合物的主要靶点。分子对接研究揭示了它们与一氧化氮合酶3（NOS-3）的血红素结合位点及乙酰胆碱酯酶的有效活性位点相互作用，与周边芳香亚位和催化三件套相互作用。最后，该化合物在模拟胃肠环境中表现出稳定性，表明有可能被吸收进入血液而没有显著的水解。这些发现突出了acefylline-eugenol 6d和acefylline-isatin 19杂交物在针对AD中涉及的多种病理机制方面的潜在治疗潜力，并为进一步开发作为该可怕疾病的潜在治疗方法提供了有希望的途径。本期刊文章受英国皇家化学学会版权所有。

In this study, we investigated the conjugation of theophylline with different compounds of natural origin hoping to construct new hybrids with dual activity against cholinergic and inflammatory pathways as potential agents for the treatment of Alzheimer's disease (AD). Out of 28 tested hybrids, two hybrids, acefylline-eugenol 6d and acefylline-isatin 19, were able to inhibit acetylcholinesterase (AChE) at low micromolar concentration displaying IC50 values of 1.8 and 3.3 μM, respectively, when compared to the galantamine standard AChE inhibitor. Moreover, the prepared hybrids exhibited a significant anti-inflammatory effect against lipopolysaccharide induced inflammation in RAW 264.7 and reduced nitric oxide (NO), tumor necrosis alpha (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) levels in a dose dependent manner. These hybrids demonstrated significant reductions in nitric oxide (NO), tumor necrosis alpha (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) levels in RAW 264.7 cells induced by lipopolysaccharide (LPS). The findings of this study were further explained in light of network pharmacology analysis which suggested that AChE and nitric oxide synthase were the main targets of the most active compounds. Molecular docking studies revealed their ability to bind to the heme binding site of nitric oxide synthase 3 (NOS-3) and effectively occupy the active site of AChE, interacting with both the peripheral aromatic subsite and catalytic triad. Finally, the compounds demonstrated stability in simulated gastric and intestinal environments, suggesting potential absorption into the bloodstream without significant hydrolysis. These findings highlight the possible therapeutic potential of acefylline-eugenol 6d and acefylline-isatin 19 hybrids in targeting multiple pathological mechanisms involved in AD, offering promising avenues for further development as potential treatments for this devastating disease.This journal is © The Royal Society of Chemistry.