Department of Pharmacy, Abdul Wali Khan University Mardan, Mardan, Pakistan.
Department of Pharmacy, Bacha Khan University, KP, Charsadda, Pakistan.
PLoS One. 2024 Oct 4;19(10):e0297398. doi: 10.1371/journal.pone.0297398. eCollection 2024.
5-Lipoxygenase (5-LOX) is a key enzyme involved in the biosynthesis of pro-inflammatory leukotrienes, leading to asthma. Developing potent 5-LOX inhibitors are highly attractive. In this research the previously synthesized isoxazole derivatives has been investigated against 5-LOX inhibitory and antioxidant in vitro assay. The compound 3 caused concentration dependent inhibition of 5-LOX with overall IC50 value of 8.47 μM. The investigated compounds C5 also exhibited good 5-LOX inhibitory effect. The IC50 demonstrated for C5 was 10.48. Among the 10 synthesized compounds, the potential 5-LOX inhibitory effect was reported for C6. The most potent compound which showed excellent free radical scavenging effect was C3 having IC50 value of 10.96 μM. The next most potent antioxidant activity was reported for C5 which non-significantly showed free radical scavenging effect. The IC50 value observed for C5 was 13.12 μM. Compound C6 also showed potent dose dependent antioxidant effect with IC50 value of 18.87 μM having percent inhibition of 91.63±0.55, 88.45±0.49, 83.53±0.45, 78.42±0.66 and 73.72±0.64 at concentration 1000-62.5 μg/mL respectively. Among the tested compounds, C6 was found most potent which showed significant 5-LOX percent inhibition assay and also reported the minimum IC50 value comparable to the reference drug. The in vitro 5-LOX enzymes inhibition assays of C5 and C3 also showed excellent percent inhibition and good potency next to C6. We concluded that amongst the investigated designed molecules the C3 was found best potent and showed significant dose dependent antioxidant activity against DPPH screening. The IC50 value reported for C3 was found good as compared to standard drug. Moreover, C5 and C6 also showed excellent free radical scavenging effect against DPPH assay. Computational methods have also been employed to explore the probable interaction model of inhibitors and enzyme active sites, and also to correlate the results of in silico and in vitro studies.
5-脂氧合酶（5-LOX）是参与炎症性白三烯生物合成的关键酶，导致哮喘。开发有效的 5-LOX 抑制剂非常有吸引力。在这项研究中，对以前合成的异噁唑衍生物进行了研究，以进行 5-LOX 抑制和体外抗氧化活性测定。化合物 3 引起 5-LOX 的浓度依赖性抑制，总 IC50 值为 8.47 μM。所研究的化合物 C5 也表现出良好的 5-LOX 抑制作用。C5 的 IC50 为 10.48。在所合成的 10 种化合物中，报道了 C6 具有潜在的 5-LOX 抑制作用。表现出极好的自由基清除作用的最有效化合物是 C3，其 IC50 值为 10.96 μM。其次是具有非显著自由基清除作用的 C5，其抗氧化活性最强，IC50 值为 13.12 μM。化合物 C6 也表现出较强的剂量依赖性抗氧化作用，IC50 值为 18.87 μM，抑制率为 91.63±0.55、88.45±0.49、83.53±0.45、78.42±0.66 和 73.72±0.64，浓度分别为 1000-62.5 μg/mL。在所测试的化合物中，C6 被发现最有效，它表现出显著的 5-LOX 抑制作用百分比测定，并且报道的最小 IC50 值与参考药物相当。C5 和 C3 的体外 5-LOX 酶抑制试验也表现出极好的抑制率和仅次于 C6 的良好效力。我们得出的结论是，在所研究的设计分子中，C3 被发现最有效，并且对 DPPH 筛选显示出显著的剂量依赖性抗氧化活性。C3 报告的 IC50 值与标准药物相比被认为是较好的。此外，C5 和 C6 也表现出对 DPPH 测定的极好的自由基清除作用。还采用计算方法探索抑制剂与酶活性位点的可能相互作用模型，并将计算和体外研究的结果相关联。
