A Biological Evaluation And Structure-Activity Relationship Study Of Hydrazone And Imide Derivatives Inhibitors Against FBA-?

Fructose-1,6-bisphosphate aldolases (EC 4.1.2.13, FBA) participate in two major carbohydrate metabolism. In gluconeogenesis, the enzyme catalyzes the process of the formation of fructose 1,6-bisphosphate (FBP) from two compounds, one is glyceraldehyde 3-phosphate (G3P) and the other is dihydroxyacetone phosphate (DHAP); while in the process of glyco lysis, FBA can catalyze the inverse reaction. Inhibiting the activity of FBA can suppress glucose metabolism blocking the whole process, which led to the death of the species.If we know more about the Class ? FBA, we will be more likely to find the environmentally friendly and efficient, highly selective inhibitors. In the study, the properties of C. albicans Class ? FBA are revealed with the use of some method such as protein expression and purification, enzymology study, inhibitor structure-activity relationship study and binding mode analysis. Also a series of novel maleimide derivatives were tested on cyanobacteria FBA-? and we test the selectivity of hit compounds.The details are summarized below:1 Ca-FBA-? is expressed using pet28a as the expression plasmid in Escherichia coli, and affinity chromatography is used to purify Ca-FBA-?. The optimized conditions for the Ca-FBA-? expression was found, and the purity of Ca-FBA-? is over 90%. The yield of the high-activity enzyme is 45 mg/L.2 We have done the Ca-FBA-II kinetics research, and study shows that the optimum temperature is 37 ?. The reactivating metal ion is K+? Zn2+, After adding a certain amount of EDTA, the enzyme activity was 0. and the optimum GPDH?TIM concentration are 2.5 U/mL?1.25 U/mL. The kinetic parameters Km, Vmax, were tested, with Km=60.5 ?M, Vmax=69.7 U.3 Screening has been carried out on Ca-FBA-? among ????? series of phenylhydrazone and hydrazone inhibitors. The final concentration of the inhibitors is 50 ?M. Then determine the IC50 of the compounds, The compound with the lowest IC50 is 1-20, and the IC50 is 300 nM showed selectivity of up to 600. The series of ?, ? have shown high inhibition rate and some of the hydrazone inhibitors show the inhibition more than 90%. The compound with the lowest IC50 is ?-6, and the IC50 is 2.7 ?M. The . above researches of Ca-FBA-? may provide new ideas to antifungal drugs and future research.4 A set of series ? compounds were tested on cyanobacteria FBA-?. Among them, compound ?-10 not only shows high Cy-FBA-? activities (IC50=1.7 ?M). The selectivity of compound IV-10 is up to 30. Thus, the compound ?-10 was selected as representative molecule, and its probable interactions with the nearby significant residues in the active site of Cy-FBA-? were elucidated by jointly using molecular docking to provide a new insight into the binding-mode of the inhibitors and Cy-FBA-?.