Synthesis And Anti-inflammatory Activity Evaluation Of Triazoleoxazine And Coumarinsoxazine Derivatives

Inflammation is defined as part of the complex biological reaction of vascular tissues to remove injury or harmful stimuli including pathogens,damaged cells or irritation.It has a close association with the occurrence of various disease.So it’s very important to develop new and effective anti-inflammatory drugs.Aiming to discover new compounds with anti-inflammatory activity,3-alkyl-6-（4H-1,2,4-triazol-4-yl）-2,4-dihydro-3H-benzo[e][1,3]oxazine derivatives and 9-alkyl-8,10-dihydrochromeno[8,7-e][1,3]oxazin-2（8H）-one derivatives were synthesized.Firstly,4-aminophenol reacted with formohydrazide to get 4-（4H-1,2,4-triazol-4-yl）phenol.Then,it reacted with formaldehyde and amine to get 3-alkyl-6-（4H-1,2,4-triazol-4-yl）-2,4-dihydro-3H-benzo[e][1,3]oxazine derivatives via mannich reaction.In addition,7-hydroxycoumarin reacted with formaldehyde and amine to get 9-alkyl-8,10-dihydrochromeno[8,7-e][1,3]oxazin-2（8H）-one derivatives by mannich reaction.And the structure of the target compounds were confirmed with ¹H-NMR、¹³C-NMR、IR and MS.The in vivo anti-inflammatory activity of the 3-alkyl-6-（4H-1,2,4-triazol-4-yl）-2,4-dihydro-3H-benzo[e][1,3]oxazine derivatives was determined using ear swelling model in mice.The study showed that 3-Heptyl-6-（4H-1,2,4-triazol-4-yl）-3,4-dihydro-2H-benzo[e][1,3]oxazine（4a）and 3-p-Tolyl-6-（4H-1,2,4-triazol-4-yl）-3,4-dihydro-2H-benzo[e][1,3]oxazine（2b）displayed higher anti-inflammatory activity（74.04%and 64.99%,respectively,at 0.5 h after intraperitoneal administration）than the reference drug Ibuprofen（62.65%）.Furthermore,the time of peak effect（TPE）for anti-inflammatory activity of 4a and 2b was observed at the 4 h after intragastric Administration.As for 9-alkyl-8,10-dihydrochromeno[8,7-e][1,3]oxazin-2（8H）-one derivatives,we use the MTT assay to evaluate the cell viabilities of RAW264.7 cells.9-（2-chlorophenyl）-8,10-dihydrochromeno[8,7-e][1,3]oxazin-2（8H）-one（B3）showed cytoactivity at a dose of 25 μg/mL.After LPS stimulation of RAW264.7 cells,the result of ELISA showed that B3 inhibited the production of TNF-a and IL-6 at concentration of 6.25 μg/mL,12.5 μg/mL and 25 μg/mL in a dose-dependent manner.Western blot assay showed that compound B3 could inhibit inflammatory responses via suppression of the NF-kB and MAPK signaling pathways.Docking study of the prepared compounds was performed for the study of the interaction between molecules and the active site of TNF-a.