Potential COX-2 inhibitors modulating NF-κB/MAPK signaling pathways: Design, synthesis and evaluation of anti-inflammatory activity of Pterostilbene-carboxylic acid derivatives with an oxime ether moiety

Abstract

In this work, a series of novel Pterostilbene–oxime ether–carboxylic acid (POC) derivatives (d1–d10, e1–e10 and 1–13) were designed, synthesized, and characterized by spectroscopic techniques. In order to further determine the absolute configuration of these compounds, one of them, compound d3, was investigated by X-ray single crystal diffraction method. d3 had a triclinic crystal with P-1 space group, and its CHCH and CHN was confirmed as E configuration. A strong hydrogen bond was formed between the hydrogen atom in CHCH moiety and the nitrogen atom in CHN moiety, which was a vital factor in the formation and stability of E configuration in the CHCH and CHN. The safety and anti-inflammatory activities of compounds (d1–d10, e1–e10 and 1–13) in vitro were evaluated. At 20 μM, compounds (d1–d10, e1–e10 and 1–13 were non-toxic and exhibited weak to strong inhibitory effects on the LPS-induced NO release. Among them, five compounds (1, 2, 7, 8 and 9) showed excellent anti-inflammatory effects with IC50 (NO) values ranging from 9.87 to 19.78 μM, as well as strong COX-2 inhibitory abilities with IC50 (COX-2) values ranging from 85.44 to 140.88 nM. Moreover, there was a rough positive correlation between their anti-inflammatory properties and the COX-2 inhibitory abilities. Compounds (1, 2, 7, 8 and 9) smoothly docked with COX-2 protein (PDB ID: 5KIR) to form stable complexes with strong hydrogen bonds, with an affinity range of −8.3 to −9.9 kcal/mol. SAR indicated that the amidation of POC at R2 position was more favorable for enhancing the compound’s biological actives than esterification. In addition, the 4-fluobenzyl substitution at R2 position of the oxime ether moiety can obviously enhance the activity of above amide derivates. Introducing acyl groups (CO(CH2)nCH3, n = 2, 4 and 6) into NH(CH2)3OH group to form ester chain is disadvantageous for activity enhancing, moreover, the longer the carbon chain, the poorer the activity. The strongest COX-2 inhibitor (IC50 (COX-2) = 85.44 ± 3.88 nM), compound 7, exerted as anti-inflammatory activities (IC50 (NO) = 9.87 ± 1.38 μM) by down-regulating the expression of COX-2 and iNOS, and modulating NF-κB/MAPK signaling pathways.