Design,synthesis And In Vitro Evaluation Of Multitargeted PPARs Agonists

Objective:The thiazolidinediones?TZDs?anti-diabetic agents targeting PPAR?are deemed as insulin sensitizer,which could control glycemic level as well as induce weight gain,edema and other side effects.Thus,TZDs were restricted in clinic.The study found that PPAR?/?dual agents could exert PPAR?-mediated hypoglycemic effect,while the inherent side effects of PPAR?agonists could be reduced via PPAR?-mediated fatty acid oxidation and other effects.In recent years,the development of PPARs multi-targeted drugs with hypoglycemic activities has become a hot spot in research institutes and pharmaceutical companies.In this paper,based on the chemical structures of known PPARs agonists and aided by computer-aided drug design?CADD?techniques,we designed and synthesized novel multi-targeted agonists with high affinity to PPARs.In addition,in vitro activities of synthesized compounds were determined.Method:?1?Based on the combination principle and the bioisosterism principle of medicinal chemistry,the polar head of bezafibrate and the hydrophobic tail of pioglitazone were integrated.Various alkoxy groups were substituted to afford carboxylic acids and tetrazole derivatives.According to the results of virtual screening and experimental feasibility analysis,we designed reasonable synthetic routes.With hydroquinone and resorcinol respectively as raw materials,the targeted combinational molecules were obtained,and their structures were confirmed by ¹H-NMR,¹³C-NMR and Mass spectrum.We utilized the cell model in PPAR?/?/?activities evaluation to determine the median effective concentration(EC₅₀)and concentration at maximum efficiency percentage through in vitro transaction assays.?2?In order to identify potential PPAR?/?dual agonists with high affinity,we utilized computational techniques to rapidly screen optimal ligands from the Specs database.?3?On the basis of selective AT₁R antagonism and PPAR?partial activity with telmisartan,we designed novel structures by modification to the structural skeleton of telmisartan.The carboxylic acid moiety,the distal benzimidazole group and the biphenyl moiety were modified.With molecular docking,ADMET prediction and other computational techniques,we expected to get potent dual ligands with selective AT₁R antagonism and PPAR?partial activity.Results:?1?In this paper,we designed 36 molecules based on the combination principle and the bioisosterism principle.According to the results of virtual screening and experimental feasibility analysis,27 combinational molecules were successfully synthesized.Their structures were confirmed by ¹H-NMR,¹³C-NMR and Mass spectrum.Preliminary in vitro PPAR?/?/?transactivation assays yielded six compounds with potential PPARs activation in comparison to positive controls.They were compound 6h with PPAR?activity,6g,10h with PPAR?activity and 6e,6l,10l with PPAR?activity.The median effective concentration(EC₅₀)values and concentration at maximum efficiency percentage were calculated accordingly.After in vitro activity evaluation,molecules 6h,10h and 6e were selected as representatives to run dynamic simulations of20 ns in the solvent with counter ions.The results indicated that the conformations of complexes and ligand itself retained relatively stable in dynamics state.The steady interaction modes for 6h,10h and 6e in the binding pockets of PPAR?,PPAR?and PPAR?respectively were validated.?2?With computational methods,seven compounds with better binding poses and beneficial pharmacokinetic properties after docking analysis,ADMET prediction were obtained via virtually screening the Specs database.Taking ZINC36517927 as representative,20 ns simulations validated the stability of docked protein-ligand complex in dynamic state.?3?10 novel structures with higher Glide scores,similar interactions and excellent pharmacokinetic profiles based on the structure of telmisartan were obtained via structural modification following by docking analysis and ADMET prediction.Molecular dynamics simulations on the best docking pose of comp#91 validated the conformational stability with both receptors throughout the course of 20 ns simulations.Conclusion:Several groups of molecules targeting PPARs were designed with molecular docking,ADMET prediction and other computer-aided drug design methods.27compounds of one group were synthesized and their structures were confirmed by¹H-NMR,¹³C-NMR and Mass spectrum.Though in vitro PPARs activities of these molecules were not good as the positive control,we believe the later optimization and pharmacological activity assays could surely provide valuable reference for the development of novel,efficient anti-diabetic drugs targeting PPARs.