Synthesis, activity, docking and dynamic simulation studies of novel pyrazolo-pyrano[2,3-d]-pyrimidine analogues as anti-diabetic agents.

• Novel pyrazolo-pyrano[2,3- d ]-pyrimidine molecular hybrids were reported. • Its structure elucidation was established via different spectroscopic techniques. • Synthesized compounds were evaluated anti-diabetic activity (in vitro) against α-glucosidase and α-amylase enzymatic inhibition. • Compound 7d showed the high potency inhibitory activity for the α-glucosidase, and α-amylase. • Binding interaction was also validated with molecular docking studies. The anti-diabetic potential of newly synthesized pyrazolo-pyrano[2,3- d ]-pyrimidine analogues (7a-j) was evaluated through comparative screening against two enzymatic targets like α-glucosidase and α-amylase. All newly prepared molecules were characterized and confirmed by 1HNMR, 13CNMR, and HR-MS analysis. The in vitro anti-diabetic activity was screened using α-glucosidase and α-amylase enzymatic inhibition using Acarbose as standard. Among these, compound (7d) exhibited the most potent anti-diabetic activity IC 50 value of 1.18 µM, and 21.63 µM for the α-glucosidase, and α-amylase inhibition respectively, surpassing the IC 50 values of 3.26 µM and 32.21 µM for the reference Acarbose. The molecules' structural study revealed that the para-position electron-withdrawing (NO 2) group were crucial to the inhibitory activity. The other hybrid scaffolds 7f, 7c, and 7 h also exhibited good inhibition compared with reference. A molecular docking study were performed for all the molecules, and based on the AutoDock vina program score, and was further considered for molecular dynamics simulations. The investigation included a look at protein stability, APO-protein dynamics, and interactions. Using molecular dynamics simulations, the overall RMSD analysis of APO protein of α-amylase (PDB: 1B2Y), and α-glucosidase (PDB: 5NN8) forms showed a stable interaction and the average RMSD value is 1.93±0.18 Å and 1.62±0.17 Å. These novel pyrazolo-pyrano[2,3- d ]-pyrimidine molecular hybrids have the potential to be useful in the treatment of diabetes. [ABSTRACT FROM AUTHOR]