Department of Pharmacy, Birla Institute of Technology & Science - Pilani, Hyderabad Campus, Hyderabad, Telangana, India.
School of Agriculture, Meiji University, Kawasaki, Japan.
Chem Biol Drug Des. 2024 Mar;103(3):e14486. doi: 10.1111/cbdd.14486.
Targeting pro-inflammatory cytokines and their production is found to be of therapeutic benefit for the regulation of inflammation in various chronic autoimmune diseases. Our continued efforts to discover small molecular-weight pro-inflammatory cytokine inhibitors resulted in identifying a novel natural lignan molecule named polonilignan, isolated from the culture broth extract of an endophytic fungus Penicillium polonicum. An in silico study (molecular docking, ADME predictions, binding free energy calculation and molecular dynamics simulation) of the polonilignan over the pro-inflammatory cytokines proteins TNF-α, IL-6 and IL-1β was performed using Schrodinger LLC software to understand the binding interactions, drug-like properties, and stability of the ligand-protein complex. Further, in-vitro testing of inhibition of TNF-α, IL-6 and IL-1β by polonilignan was carried out using ELISA and RT-PCR on LPS-induced RAW 264.7 cell lines along with the testing of nitrite production effect (Griess assay) and cytotoxicity (MTT) analysis. Under the computational study, polonilignan revealed good docking scores, binding interactions, and stability under MDS and desirable in silico ADME results over the proteins TNF-α, IL-1β and IL-6. Poloniligan showed significant inhibition of IL-1β, IL-6 and TNF-α with IC values of 2.01 μM, 6.59 μM and 42.10 μM, respectively. Also, it reduced the translocation of the NF-κB subunit p65 to the nucleus (confocal microscopy). The mRNA expression levels of pro-inflammatory markers IL-1β, TNF-α and IL-6 levels were lowered significantly (p < .001) by the compound, and the diminution was higher with IL-1β. Further, the lignan was non-cytotoxic and effective in attenuating nitrite release (IC 48.56 μM). Thus, polonilignan has been identified as a new pan-cytokine and NO inhibitor, it is recommended to optimise a method for the synthesis of this small molecular weight lignan and explore its pharmacokinetic characteristics, toxicity and therapeutic effect under various chronic inflammatory disease models.
针对促炎细胞因子及其产生的靶向治疗被发现对各种慢性自身免疫性疾病中的炎症调节具有治疗益处。我们持续努力发现小分子促炎细胞因子抑制剂，结果鉴定出一种新型天然木脂素分子，名为波罗尼烷，从内生真菌 Penicillium polonicum 的发酵液提取物中分离得到。使用 Schrödinger LLC 软件对波罗尼烷进行了针对促炎细胞因子蛋白 TNF-α、IL-6 和 IL-1β 的计算研究（分子对接、ADME 预测、结合自由能计算和分子动力学模拟），以了解配体-蛋白复合物的结合相互作用、类药性和稳定性。此外，通过 ELISA 和 LPS 诱导的 RAW 264.7 细胞系上的 RT-PCR 对波罗尼烷抑制 TNF-α、IL-6 和 IL-1β 的作用进行了体外测试，同时还进行了亚硝酸盐产生效应（Griess 测定）和细胞毒性（MTT）分析。在计算研究中，波罗尼烷在 MDS 下显示出良好的对接评分、结合相互作用和稳定性，以及对 TNF-α、IL-1β 和 IL-6 蛋白的理想的计算 ADME 结果。波罗尼烷对 IL-1β、IL-6 和 TNF-α 的抑制作用显著，IC 值分别为 2.01 μM、6.59 μM 和 42.10 μM。此外，它还减少了 NF-κB 亚基 p65 向核内的易位（共聚焦显微镜）。该化合物显著降低了促炎标志物 IL-1β、TNF-α 和 IL-6 的 mRNA 表达水平（p <.001），并且 IL-1β 的降低幅度更高。此外，该木脂素无细胞毒性，有效抑制亚硝酸盐释放（IC 48.56 μM）。因此，波罗尼烷已被鉴定为新型的泛细胞因子和 NO 抑制剂，建议优化该小分子木脂素的合成方法，并在各种慢性炎症性疾病模型中探索其药代动力学特征、毒性和治疗效果。
