Inhibitory Activity And Regulation Mechanism Of GSH、Nitrovinylbenzoic Acid Ester、Maleimide Compounds On Hu-FBPase

Diabetes is a metabolic disease characterized by high blood sugar,and the incidence of type 2 diabetes(T2DM)is growing rapidly around the world.It is one of the most common,most expensive and deadly chronic diseases in the world.Human liver fructose-1,6-bisphosphatase(Hu-FBPase)is a key regulatory enzyme in the gluconeogenesis pathway,which can achieve hypoglycemic effect by inhibiting Hu-FBPase,and is considered to be an important target for the treatment of type 2 diabe-tes.Hu-FBPase is a homotetramer that contains a substrate binding site and an AMP allo-steric site in each monomer.Many of the inhibitors of Hu-FBPase are currently designed for allosteric sites,and most of them are exogenous small organic molecule inhibitors,and most of the binding methods are non-covalent binding.However,AMP is also in-volved in important physiological regulation of the human body.Therefore,inhibitors designed for AMP sites will bring about some problems such as poor selectivity and large side effects.The FBP site is hi ghly hydrophilic and it is difficult to design inhibitors.Therefore,we want to fin dan inhibitor that is less toxic and binds to new sites.This article mainly carried out the following work:1.The inhibitory activity of human small molecules on Hu-FBPase were test ed.It was found that human small molecule glutathione(GSH)can inhibit Hu-FB Pase.The specific research contents are as follows:(1)The IC50 of GSH is 1.64±0.14 mM,and it is recommended to bind to t he new site of the Hu-FBPase central cavity site.(2)GSH and AMP have a synergistic effect,and GSH can increase the inhib itory activity of AMP by 12 times.At the same time,GSH also has a synergistic effect with the allosteric site inhibitor R03,and GSH can increase the inhibitory effect of R03 by 39 times.2.The nitrostyrene compounds were synthesized in the early stage of this res earch group,and the binding sites of the compounds were determined.This class of compounds was found to inhibit the activity of Hu-FBPase by forming a coval ent bond with C128.According to the modification mode with Hu-FBPase,nitrovi nylphenyl benzoate compounds was synthesized.The most potent compound 107((E)-3-(2-nitrovinyl)phenyl 3-phenoxybenzoate),120((E)-3-methoxy)-5-((3-(2-Nitrovi nyl)phenoxy)carbonyl)benzoic acid),124((E)-3-(2-nitrovinyl)phenylquinolin-5-carboxy lie acid),the IC50 of these esters were 0.14±0.01 μM,which was 25 times higher than before the modification.Compound 124 has a good hypoglycemic effect on primary liver cells.3.The maleimide compounds having an electrophilic group were synthesized according to the principle of covalent bonding.The inhibitory constant IC 50 of 39 maleimide compounds against Hu-FBPase were determined,and the compound 212 with the best inhibitory effect(N-(3-bromo-2,5-dioxo-2,5-dihydro-)was determined.The IC50 of 1H-pyrrol-1-yl)-4-chlorobenzamide was 0.11±0.02 μM.