| Endogenous H2S,as the third novel gas signal molecule similar to NO and CO,has extensive pharmacological effects such as anti-inflammatory,gastrointestinal protection,dilation of blood vessels,lower blood pressure,and myocardial protection in vivo,but pure H2S gas has The disadvantages of uncontrollable dose and high dose toxicity limit their clinical application.The exogenous H2S release molecule,as a solidified form of H2S,can release H2S in a controlled and quantitative manner in vivo,and has the pharmacological effect of endogenous H2S,becoming one of the hot spots in the field of chemistry and biology.Non-steroidal anti-inflammatory drugs（NSAIDs）,as the most common anti-inflammatory drugs,exert severe gastrointestinal adverse reactions while exerting their good anti-inflammatory effects,and long-term use of elevated average arterial pressure,based on new drugs.In the research and development strategy,we spliced NSAIDs and exogenous H2S releasing molecules to expect that the synthesized compounds would have synergistic anti-inflammatory effects while also reducing the gastrointestinal adverse reactions of NSAIDs and raising the risk of mean arterial pressure.In this paper,three classes of non-steroidal anti-inflammatory H2S-releasing molecules were synthesized and characterized.The H2S release characteristics,cell and animal toxicity,and anti-inflammatory activity were studied and evaluated.The main research contents include the following:aspect:（1）H2S release characteristics:The H2S release ability of all the target compounds was measured in the presence of TCEP or cysteine by the H2S release ability of the UV-visible spectrophotometric compound.The results show all compounds were fast H2S-releasers,and their half-lives were in range of 0–20 min.And under the same conditions,Compound 12 has the highest H2S release among all compounds.（2）Cytotoxicity studies:All compounds except Compounds 1 and 2 showed extremely low toxicity to both LO2 and HepG2 cells,with the IC50 values of most compounds exceeding 800 mM.Compounds 1 and 2 have stronger antiproliferative activity against both cells,but they are less toxic to LO2 cells than HepG2 and show some cell selectivity.（3）Animal Developmental Toxicity Study:Based on cytotoxicity,zebrafish embryos were used to evaluate the developmental toxicity of the compounds.The results showed that all tested compounds 2,12 and 18 had an impact on the mortality,hatchability,and the spontaneous movement of zebrafish embryos and resulted in embryo malformations;the compounds were dose-dependent on embryonic and larval zebrafish toxicity.（4）Animal Physiological Toxicity Study:Based on cytotoxicity,rats were used to assess the developmental toxicity of the compounds.The results showed that after continuous long-term dose administration in rats,compounds 2,12,and 15 reduced the leukocyte content in rat blood,increased the protein and leukocyte content in urine,indicating that the physiological state of rats was affected;2,12 has an effect on the physiological morphology and function of rat liver,and has a severe effect on the function and physiological form of the kidney.（5）Anti-inflammatory activity:The target compounds all have anti-inflammatory effects and can reduce the level of nitrite in macrophages;its anti-inflammatory effects are related to the ligand species and H2S release rate.In the anti-inflammatory process,the target compounds reduce the content of proinflammatory cytokine TNF-α,increase the expression of anti-inflammatory factors IL-10 and HO-1,and increase the expression of COX-2 through the immune regulatory system.（6）Gastrointestinal tract protective effect:After a continuous administration of ibuprofen and compound 2 in rats for a long period of time,we stained by HE and observed the rat’s gastrointestinal tract by microscope.The results showed that the compound 2 group was congested in the stomach.Symptoms were significantly reduced compared with the ibuprofen group and no swelling of the gastric cells occurred;symptoms of intestinal mucosal congestion were also significantly reduced.This shows that compound 2 may have a certain improvement on the gastrointestinal side effects of ibuprofen.（7）Lowering blood pressure and myocardial protection:We selected spontaneously hypertensive rats（SHR）to study the effect of compounds on blood pressure in a hypertension study model.The results showed that compounds 2,12,and 15 had a good blood pressure lowering effect on spontaneously hypertensive rats（SHR）and had a concentration-dependent effect.The duration of blood pressure reduction was similar to that of nifedipine and captopril.However,its antihypertensive effect was weaker than nifedipine and captopril;and compounds 2,12 and 15 had a good effect on myocardial damage in spontaneously hypertensive rats（SHR）. |
