Design, Synthesis, and Biological Activity of Tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidine Derivatives as Anti-Inflammatory Agents.

Yuan Zhang,Di Chen,Lu Luo,Kaiqi Wu,Handeng Lv,Faqing Ye,Suwei Pan,Guoliang Shen,Chao Han

doi:10.3390/molecules22111960

Abstract

We designed and synthesized 26 prototype compounds and studied their anti-inflammatory activity and underlying molecular mechanisms. The inhibitory effects of the compounds on the production of nitric oxide (NO), cytokines, inflammatory-related proteins, and mRNAs in lipopolysaccharide (LPS)-stimulated macrophages were determined by the Griess assay, Enzyme linked immunosorbent assay (ELISA), Western blot analysis, and Reverse transcription-Polymerase Chain Reaction (RT-PCR), respectively. Our results indicated that treatment with A2, A6 and B7 significantly inhibited the secretion of NO and inflammatory cytokines in RAW264.7 cells without demonstrable cytotoxicity. It was also found that A2, A6 and B7 strongly suppressed the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase enzyme COX-2, and prevented nuclear translocation of nuclear factor κB (NF-κB) p65 by inhibiting the degradation of p50 and IκBα. Furthermore, the phosphorylation of mitogen-activated protein kinase (MAPKs) in LPS-stimulated RAW264.7 cells was significantly inhibited by A2, A6 and B7. These findings suggest that A2, A6 and B7 may operate as an effective anti-inflammatory agent through inhibiting the activation of NF-κB and MAPK signaling pathways in macrophages. Moreover, rat paw swelling experiments showed that these compounds possess anti-inflammatory activity in vivo, with compound A6 exhibiting similar activities to the reference drug Indomethacin.

Highlights

Activated macrophages play crucial roles in the initiation and maintenance of inflammation.Following activation by inflammatory stimuli such as lipopolysaccharide (LPS), macrophages secrete a number of potent bioactive inflammatory mediators, including nitric oxide (NO), prostaglandins (PGs), leukotrienes (LTs), and cytokines such as interleukin-1β (IL-1β), IL-6, and tumor necrosis factor α (TNF-α), that contribute to the activation of mitogen-activated protein kinase (MAPK)and nuclear factor κB (NF-κB) [1,2,3,4,5,6,7,8,9,10]
We attempted to introduce a carbonyl in place of the amino group, and to modify the 2, and positions the tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidine system
The anti-inflammatory effects of these compounds were evaluated in a variety of in vitro and in vivo assays, including the inhibition of NO, inducible NO synthase (iNOS), and COX-2 production; pro-inflammatory cytokine secretion; and mRNA expression

Summary

Introduction

Activated macrophages play crucial roles in the initiation and maintenance of inflammation.Following activation by inflammatory stimuli such as lipopolysaccharide (LPS), macrophages secrete a number of potent bioactive inflammatory mediators, including nitric oxide (NO), prostaglandins (PGs), leukotrienes (LTs), and cytokines such as interleukin-1β (IL-1β), IL-6, and tumor necrosis factor α (TNF-α), that contribute to the activation of mitogen-activated protein kinase (MAPK)and nuclear factor κB (NF-κB) [1,2,3,4,5,6,7,8,9,10]. Following activation by inflammatory stimuli such as lipopolysaccharide (LPS), macrophages secrete a number of potent bioactive inflammatory mediators, including nitric oxide (NO), prostaglandins (PGs), leukotrienes (LTs), and cytokines such as interleukin-1β (IL-1β), IL-6, and tumor necrosis factor α (TNF-α), that contribute to the activation of mitogen-activated protein kinase (MAPK). As an important inflammatory mediator, the paradoxical role of NO in the pathogenesis of inflammation generally depends upon concentration [11]. Sustained overproduction of NO by iNOS is believed to be Molecules 2017, 22, 1960; doi:10.3390/molecules22111960 www.mdpi.com/journal/molecules detrimental to the host and is associated with the pathogenesis of a variety of inflammatory disorders [13]. Pharmacological interference with NO production is appreciated as a associated with the pathogenesis of a variety of inflammatory disorders [13]. Interference with NO production isintervention appreciated asina inflammatory promising strategy of therapeutic interventionwhile in theinflammatory major role ofdiseases housekeeping cyclooxygenase enzyme

Methods

Results

Conclusion