Semi-Quantitative Studies On The Relationship Among Anti-cancer Activities,AMPK Pathway And Chemical Structures Of Biguanides

BACKGROUND:AMP-activated kinase(AMPK)is a major cellular energetic sensor,which controls metabolic and catabolic pathways in a cell.AMPK in recent years has become a target therapy in tumor prevention and control as studies have shown that AMPK suppresses cell proliferation in tumor cells.This thesis is into two parts;1)Anti-cancer properties and mechanism of Proguanil.2)Quantitative Structural Analysis Relationship(QSAR)of Phenformin derivatives as activators of AMPKMETHODS:i)UMUC3,T24 and A549 were used in this study.Cell proliferation was assessed by MTT assay.Clonogenic and migration assays were performed to assess the effects the proguanil on cells colony formation and cell migration respectively.The effects of proguanil on the AMPK pathway were determined by western blotting.ii)2D models were built for 54 phenformin derivatives with growth inhibition of 50%from literature.Compounds were divided into two(2)groups;training set and test set 40 compounds and 14 compounds respectively.QSAR study analysis was performed on the training set and the model was used on the test set.Molecular docking was performed with the test set of compounds and molecular descriptors were determined by 2D QSAR studies using the Lipinski rule-of-five of which all the compounds satisfied all the rules.RESULTS:i)Proguanil inhibited cell proliferation in a dose-dependent manner in all the three cell lines used in this project.Proguanil inhibited the formation of cells colonies in dose-dependant manner in all the cell lines.It also inhibited the migration cells.Western blot experiment showed that proguanil is able to activate AMPK.ii)All compounds in the training set satisfied Lipinski rule-of-five.A linear correlation plot resulted from the regression analysis for the training set of 40 compounds showed a good linearity[$PRES = 0.838(pIC50)+ 0.795],with RMSE = 0.167 and R2 = 0.838 correlation regression of the test set also showed a good linear relation set[$PRES =1.013(pIC50)-0.072],with RMSE = 0.169 and R2 = 0.848.The highest docking score was P2(-10.066 Kcal/mol)and the lowest was P21(-12.262 Kcal/mol)after docking the test set of 14 compounds.CONCLUSION:Proguanil has anti-proliferative and anti-metastatic effect in bladder cancer cells UMUC3 and T24 and on lung cancer cell A549.IC 50 values were lower that Metformin compared to previous studies.Suggesting that proguanil may be more potent that the Metformin and also a drug already in the market,it is worthy of further research into its anti-cancer properties.P 21 entry of the phenformin derivatives shows the least docking score and this study predicted that phenformin derivatives in this study could be used as effective and safer therapeutics to control cancer by activating AMPK.