Estudo in silico de alvos terapêuticos no tratamento do Diabetes Mellitus

Abstract

Diabetes Mellitus (DM) is one of the most prevalent metabolic disorders in the world, with a growing search for new therapeutic agents such as food peptides. This search can be optimized using bioinformatics tools. Thus, the aim of the present study was to analyze potential therapeutic targets for glycemic control. A systematic review (SR) was developed, following the recommendations of the Preferred Reporting Items Checklist for Systematic Review and Meta-Analysis (PRISMA), according to the protocol registered in PROSPERO (CRD42022353808) and published. In the SR, studies that met the PECo strategy (Problem, Exposure, Context) were included. The databases used were: Medline (PubMed); Web of Science; Scopus; Base; ScienceDirect; Virtual Health Library (VHL). A total of 1878 articles were identified, including 20 articles referring to original in silico studies that used therapeutic targets for the treatment of DM and that validated the use of these targets in vivo. The risk of bias was assessed using a checklist based on Strengthening the reporting of empirical simulation studies (STRESS). It was observed that DPP-IV, PPARγ and GLUT4 were the most frequently used therapeutic targets in the studies included in the SR. Also through in vivo studies, the validation of the previously mentioned therapeutic targets was verified, in addition to Akt, GLP-1, α-amylase, GIP, IRS1 and GSK-3, analyzed in silico, the importance of computational simulation was verified as a useful tool in tracking and prior selection. For the in silico study, the tamarind seed trypsin inhibitor (ITT) was obtained by trypsin-sepharose 4B-CNBr affinity chromatography and characterized by antitryptic activity, protein quantification and SDS-PAGE gel. The ITT was hydrolyzed in vitro to monitor enzymatic susceptibility and selection of enzymes for cleavage in silico, using the protocol adapted from INFOGEST, simulating the three stages of digestion and monitoring the hydrolysis pattern by SDS-PAGE gel. Based on this information, the theoretical model of the purified tamarind seed trypsin inhibitor (TTIp 56/287) was cleaved in silico, being selected for simulation by molecular dynamics, Peptideotripquimo59 for presenting greater potential for interaction with the insulin receptor (IR) (PDB ID 4OGA), with a docking score of -175,53. Peptideotripchyme59 showed affinity and stability with the IR, reaching equilibrium at the beginning of the simulation and increased stability when added to the complex formed by insulin/IR, with synergistic behavior, considering the EPI. In addition, the amino acid residues Arginine at position 16 (- 209.07 kJ mol-1), Threonine at position 1 (-148.54 kJ mol-1) and Valine at position 2 (-94.53 kJ mol-1) were identified. -1) as the ones that most interacted, with this site of interaction with the IR being in a different region compared to that of insulin. Through SR, the diversity of targets that can be used for the treatment of DM were analyzed in silico and validated in vivo, contributing to the discovery of new allies for the treatment of DM. In addition, Peptideotripchymo59 proved to be an insulin mimetic molecule, and may act as a possible candidate for the treatment of type 1 and 2 DM.