Toxicity,biological Activity And Pharmacokinetics Of Thiophosphoramide H₂S Donors

H₂S is considered as the third gas signal molecule after CO and NO,and its gas form is inconvenient for clinical use.It is a practical strategy to use H₂S donor instead of H₂S gas.Among H₂S donors,thiophosphoramid derivatives are a kind of H₂S donors with low cytotoxicity.They not only release H₂S,but also exhibit good biological activities,such as anti-inflammatory and myocardial protection.However,these compounds have not been studied and reported for their toxicity,pharmacokinetics and other properties.Therefore,in order to evaluate whether such compounds may be candidate drugs for clinically relevant diseases,the subject has synthesized Based on this,their H₂S release ability,toxicity,biological activity and pharmacokinetic properties were studied in detail.The main results of the research include the following aspects:?1?H₂S release ability of compounds and its influencing factorsH₂S release was detected using a fluorescent probe method.The results show that all compounds release H₂S at room temperature,and most H₂S release curves conform to the first-order kinetic equation.For the first series of compounds,the 1-9 half-life is about 30 min.For the second series of compounds 10-18,except for compound 14,the half-life of other compounds is about 3.5 min.When the temperature increased,the maximum release of the first series of compounds increased slightly,but its half-life did not change significantly.For the second series of compounds,the maximum release also increased slightly,and the half-life was shortened,while Compound 14 remained unchanged.When the pH changes,the release half-life of the first series of compounds 1 and 7 does not change much,but the lower the pH value of the second series of compounds,the faster the H₂S release rate.The higher the pH value,the slower the release rate of H₂S,but the change in pH did not affect the maximum release amount of the two series of H₂S donors.?2?H₂S release ability of compounds in organs and tissuesLC-MS was used to detect the product of the compound after H₂S was released in PBS and serum,and then the liver,heart,and kidney homogenates of Wistar rats were used to simulate the process of H₂S release in vivo.The results show that the compounds release H₂S at different rates in different tissues,but the H₂S release mechanism is the same.?3?PharmacokineticsMale WISTAR rats were selected as animal models and analyzed by LC-MS.50minutes after the administration of Compound 1,the plasma concentration of Compound 1 reached a maximum,and then decreased sharply with time.After 12hours,Compound 1 was not detected in rat plasma,and was metabolized to other substances or excreted from the body.For compound 18,the plasma concentration reached a maximum after 100 minutes of administration.After 6 hours,no compound18 was detected in the plasma,which was metabolized to other substances or excreted from the body.In contrast,Compound 1 absorbed faster than Compound 18,and Compound 1 metabolized more slowly than Compound 18.?4?ToxicityAll 18 compounds have lower inhibitory activity on the growth of HeLa,A549,HePG2,MCF-7,HT-29 and W138 cell lines.Within the set dose range,the mice had no obvious toxicity;however,after 14 days of continuous administration,the rats'liver,kidneys and spleen showed different degrees of damage.Secondly,zebrafish embryos were used as a model.By tracking and monitoring their hatchability and survival rates,the results showed that the compounds at low concentrations were safe for the development of zebrafish embryos,and they were slightly neurotoxic at higher concentrations.Compounds exceeding 1?M cause embryo deformities.Compared with compound 1,compound 18 is more toxic.?6?Anti-inflammatory activityIn experiments using macrophage RAW264.7 as a model to test and verify the anti-inflammatory activity of compounds,all compounds reduced the nitrite content in macrophage RAW264.7,and all reduced TNF in a concentration-dependent manner-?levels and increased IL-10 levels showed good anti-inflammatory activity,with Compound 18 having the best anti-inflammatory activity.?7?Myocardial protectionThe H9c2 model of rat cardiomyocytes treated with H₂O₂ was used to test the myocardial protective effect of the compound,and it was found that the compound can exert myocardial protective activity at a specific concentration.The tested compounds can affect the content of SOD and MDA in H9c2 cells,among which compound 18 has a better activity.