Designed and Synthesized Novel Tripeptides Targeting Diabetes and its Related Pathologies

Abstract

In diabetes and its associated pathologies, such as neuropathy, glycation, α-amylase, and α-glucosidase play a crucial role. Glycation, a non-enzymatic process, causes the accumulation of protein adducts in tissues over time, interfering with normal metabolism. The urgent need for anti-glycating agent(s) that demonstrate inhibitory effects on α-amylase and α-glucosidase to prevent the formation of these proteins adducts prompted our research. This study designed a tripeptide with sequence RWW as a glycation, α-amylase, and α-glucosidase inhibitor utilizing in silico approaches and the glycation-prone Human serum albumin (HSA) regions. Further, virtual screening using HEX, AutoDock Vina, PatchDock/FireDock, and molecular dynamics was done to testify to the stability of the designed peptide docked with HSA. Eventually, this peptide was chemically synthesized and examined using HPLC, NMR, and mass spectrometry. RWW displayed remarkable inhibitory potency against α-glucosidase (IC50- 97 mM), α-amylase (IC50- 90 mM), and glycation (84%).Furthermore, in-vivo testing of RWW (63.81 µg/kg body weight) in diabetic mice compared to glimepiride (10 mg/kg body weight) revealed significant regulation of blood glucose levels, HbA1c, and serum lipid profiles, including TC, LDL-C, VLDL-C, TG, HDL-C, TC/HDL-C, LDL-C/HDL-C, and Non-HDL-C, along with ASL and ALT. After receiving RWW treatment 21 days, the kidney tissues underwent histological testing. It was found that Bowman's space and glomerulus diameters shrank, and the tubule epithelium showed improvement. Moreover, liver weight was also found to be relatively improved compared to diabetic mice. Due to its anti-hyperglycemic activity, RWW may be suggested as a potential anti-diabetic prodrug which can also be useful in inhibiting diabetic neuropathy and nephropathy.

Note:
Funding declaration: The authors extend their appreciation to the King Salman Centre for Disability Research for funding this work through Research Group no KSRG-2022-012.

Conflict of Interests: The authors declared that none had conflict in submitting and publishing this research article.

Ethical Approval: Ethical approval for the study was granted by the Institutional Animal Ethics Committee of Integral University (Letter number–IU/Pharm/Ph.D/IAEC/15/13), and the experiments were conducted in accordance with established guidelines.

Keywords: Glycation, Tripeptide, RWW, α-amylase, α-glucosidase