Design,Synthesis And Biological Evaluation Of Hesperetin Derivatives As Potent Anti-inflammatory Agent

Hesperetin,the main active ingredient is the Rutaceae orange peel of citrus fruits of plants belonging to flavanone in pale yellow needle crystal or powder.It's reported to have a variety of pharmacological activities such as anti-inflammation,anti-oxidation,hypolipidemic effect,anti-tumor effect,cardiovascular protection and immune system regulation.In addition,hesperetin can also be used for other medications such as health care series.The best-selling health products Amway's series of products have added a certain amount of hesperetin.There is also the United States in recent years is very popular health products 50,its main component content is also hesperetin.In Japan,some foods are also associated with the components of hesperetin,which is mainly used as an additive added to the vitamin p to exert its functional effects.In addition,in China,hesperetin is mainly used as a drug substance,and related drug applications include stomach,sputum,antitussive,dispelling wind,diuretic,non-reversing and stopping stomach pain.Therefore,hesperetin has attracted the attention of botanists and pharmacologists.Based on previous studies and literature surveys,we know that hesperetin can reduce ventilator-induced injury by inhibiting the activation of NF-?B signaling pathway and inhibiting the production of tumor necrosis factor(TNF)-? and interleukin6(IL-6).Acute lung injury.In another study,the authors introduced the amino group Mannich modification at the C-6 position according to their structural characteristics.When the amines introduced were isopropylamine,isobutylamine,and ethylpiperazine,the results showed that the compounds showed Better inhibition of TNF-? and IL-6expression.Bozena Kaminska found that MAPK signaling pathway can be targeted for anti-inflammatory molecular therapy,so we envisage whether hesperetin and its derivatives can exert special anti-inflammatory effects through the dual effects on NF-?B and MAPK signaling pathways.However,the bioavailability of hesperetin itself is relatively low,only a large dose will play a role,hesperetin as a direct treatment of drug expectations there is still a long way to go.Currently,there are few compounds that can be developed as pharmaceuticals,and there is a lack of research on biological activities and mechanisms.Therefore,the further modification of the structure of hesperetin has great significance for the study of the drug-forming properties of hesperetin.In this paper,hesperetin was used as raw material.The purpose was to introduce a derivative with an amide side chain substitution at position C-7.The synthesis of the compound firstly utilizes the substitution reaction and ethyl bromocrotonate to be hydrolyzed under acid heat to obtain substituted hesperitin,which then undergoes amidation reactions with primary and secondary amines.This section synthesizes a total of 29 tertiary series.Hesperetin derivatives include hesperetin compounds having heterocyclic amines,linear amines,and aromatic amines at the end of the side chain.The structure of the target compound is confirmed by mass spectrometry and NMR.The mouse model of peritoneal macrophages(RAW264.7)was used to study the anti-inflammatory activity of derivatives in vitro and the structure-activity relationship of the compounds was summarized.The effects of different concentrations of compounds on the concentrations of NO,TNF-?,and IL-6 in supernatants of RAW264.7 cells were determined using Griess reagent(Nitric Oxide Assay Kit)andELISA.The results showed that the compound 7-O-crotonyl N-ethylpiperazinyl hesperetin(5b)has the highest anti-inflammatory activity and has further research value.Based on the results of anti-inflammatory activity screening,the mechanism of the antiinflammatory effects of the derivatives was preliminarily investigated.It was found that Compound 5b can produce anti-inflammatory effects through affecting the two signaling pathways of NF-?B and MAPK.Specific research includes the following three parts:1.Synthesis of hesperetin derivatives: Hesperetin was used as a raw material to introduce an amide group into hesperetin C-7,and the product passed hesperetin and ethyl bromocrotonate in a primary amine and secondary The amine or fatty amine reaction is obtained.2.Screening for anti-inflammatory activity.Mouse peritoneal macrophages(RAW264.7)were cultured in vitro and given different concentrations of compounds.Different concentrations of compounds were measured using the Griess reagent Griess reagent nitric oxide assay kit.The expression of NO in supernatants of RAW264.7 cells was examined by ELISA to determine the effect of TNF-? and IL-6 concentrations in the supernatant.3,Western blot method to explore the possible mechanism of anti-inflammatory activity of compounds: Compounds can inhibit the expression of downstream related proteins(p-I?B?,p-p65)through NF-?B signaling pathway or inhibit downstream related proteins through another MAPK signaling(ERK),p38)Expression reaches antiinflammatory activity.