Targeting the Glucokinase-Glucokinase Regulatory Protein Axis: Pharmacophore-Based Identification of Novel Antidiabetic Small Molecules.

Diabetes is the most prevalent metabolic disorder distinguished by increased blood glucose levels. Unfortunately, none of the marketed treatments can cure the ailment. Glucokinase has recently emerged as a novel target acting in both the liver and the pancreas. The activity of this novel protein in the liver is regulated by a protein called glucokinase regulatory protein (GKRP). The study aimed to identify novel GKRP modulators and evaluate their potential as antidiabetic agents using a comprehensive approach incorporating in silico, in vitro, and in vivo methods. In the quest for novel GKRP modulators, pharmacophore models were developed using Hypogen and HipHop methodologies. The optimal pharmacophore hypothesis, featuring 1HBA, 1HY, 1HBD, and 1RA, demonstrated a root mean-square deviation of 0.88 Å and a high correlation coefficient of 0.88. Fisher's randomization and Cat Scramble tests confirmed the statistical validity of the models, with a 95% confidence level. The validated pharmacophore model was employed in a virtual screening of the NCI database, resulting in the retrieval of key hits, including NCS 1972 and NCS 80683. These compounds were subjected to docking studies and in vitro enzyme-based GKRP modulatory assay, revealing favorable binding interactions with the GKRP active site and IC50 of 1.60 and 3.04 nM, respectively. In vivo evaluations showed that NCS 1972 (2.5 mg/kg) significantly improved lipid profiles, reduced liver hypertrophy and adiposity, and enhanced body mass index, 0.27 ± 0.02 g/mm, 0.21 ± 0.12g/mm, 0.70 ± 2.82 g/cm2 respectively, compared to the HFD-STZ group. Histopathological analyses demonstrated substantial cellular restoration in pancreatic and liver tissues, with NCS 1972 exhibiting superior efficacy over NCS 80683. Additionally, gene expression studies revealed that both compounds corrected HFD-STZ-induced dysregulation of glucose metabolism and inflammatory markers. These findings underscore the significant therapeutic potential of NCS 1972 in mitigating diabetes, suggesting its promise as a novel antidiabetic agent.