Chandrasekaran Vishnupriya, Samykannu Gopinath, Natarajan Jeykumar, Rangasamy Kavitha
Biomedical and Microbial Research Laboratory, Department of Biochemistry, Bharathiar University, Coimbatore, Tamil Nadu 641 046, India.
Department of Plant Molecular Biology and Bioinformatics, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu 641 003, India.
Comput Biol Chem. 2025 Dec;119:108541. doi: 10.1016/j.compbiolchem.2025.108541. Epub 2025 Jun 5.
Non-steroidal anti-inflammatory drugs (NSAIDs) often cause adverse effects, leading to increased interest in the identification of natural compounds with fewer side effects. This study reports the purification and characterization of an alkylated flavonoid, Brevi-inflammin, a novel marine actinobacteria-derived compound that contains anti-inflammatory activity from Brevibacterium casei VRK 1. Using the one-pot synthesis method, the alkylated flavonoid compound Brevi-inflammin was synthesized and purified using semi-preparative reverse-phase HPLC. Further, it is characterized through multiple spectroscopic techniques, including UV-visible spectroscopy, FTIR, HPLC NMR, and UPLC-MS/MS. To assess the anti-inflammatory potential of Brevi-inflammin, the cytotoxic assessment and nitrous oxide inhibitory activity were performed. Further, molecular docking and molecular dynamics simulations were used to evaluate the binding affinity and stability of Brevi-inflammin with a COX-2. Spectroscopic results revealed Brevi-inflammin as an alkylated flavonoid compound, evidenced by its maximum absorption at 230 nm, characteristic aromatic ring structures (CH and CH stretching vibrations), and alkylation signature at 1652 cm. Additionally, HPLC results highlighted Brevi-inflammin as a flavonoid compound. Structural verification through NMR studies confirmed Brevi-inflammin as an alkylated flavonoid. UPLC-MS/MS results confirmed its molecular mass as 279.05 m/z. Cytotoxicity studies proved the compound's safety up to a concentration of 250 μg/mL. The inhibition of nitrous oxide production was observed at a concentration of 1000 µg/mL. Molecular dynamic simulations revealed strong binding interactions between Brevi-inflammin and COX-2, mediated through hydrogen bonds, hydrophobic interactions, and van der Waals forces. Combining experimental and computational approaches, this comprehensive characterization establishes that Brevi-inflammin could be a promising candidate for anti-inflammatory applications.
非甾体抗炎药（NSAIDs）常常会引发不良反应，这使得人们对鉴定副作用较少的天然化合物的兴趣日益增加。本研究报告了一种烷基化黄酮类化合物——短杆菌抗炎素（Brevi-inflammin）的纯化及特性，它是一种源自海洋放线菌的新型化合物，具有来自酪酪短杆菌VRK 1的抗炎活性。采用一锅合成法，通过半制备反相高效液相色谱法合成并纯化了烷基化黄酮类化合物短杆菌抗炎素。此外，还通过多种光谱技术对其进行了表征，包括紫外可见光谱、傅里叶变换红外光谱、高效液相色谱核磁共振以及超高效液相色谱-串联质谱。为评估短杆菌抗炎素的抗炎潜力，进行了细胞毒性评估和一氧化氮抑制活性检测。此外，还利用分子对接和分子动力学模拟来评估短杆菌抗炎素与环氧化酶-2（COX-2）的结合亲和力和稳定性。光谱结果表明短杆菌抗炎素是一种烷基化黄酮类化合物，其在230nm处有最大吸收、具有特征性芳香环结构（CH和CH伸缩振动）以及在1652cm处有烷基化特征峰可作为证据。此外，高效液相色谱结果突出显示短杆菌抗炎素是一种黄酮类化合物。通过核磁共振研究进行的结构验证证实短杆菌抗炎素是一种烷基化黄酮类化合物。超高效液相色谱-串联质谱结果确认其分子量为279.05m/z。细胞毒性研究证明该化合物在浓度高达250μg/mL时是安全的。在浓度为1000μg/mL时观察到一氧化氮生成受到抑制。分子动力学模拟揭示了短杆菌抗炎素与COX-2之间通过氢键、疏水相互作用和范德华力介导的强结合相互作用。综合实验和计算方法，这一全面表征表明短杆菌抗炎素可能是抗炎应用的一个有前景的候选物。
