A novel family of small molecule HIF-1 alpha stabilizers for the treatment of diabetic wounds; an integrated in silico, in vitro, and in vivo strategy.

Hypoxia-inducible factor-1 alpha (HIF-1α) is a crucial regulator of wound healing, which includes epithelialization, angiogenesis, granulation, tissue development, and wound contraction. Even though diabetic wounds are hypoxic, HIF-1α levels are decreased during healing. Diabetic wound healing necessitates the modulation of hypoxia-induced responses by VHL-HIF-1α protein-protein inhibition. Our proposed hypothesis is to increase HIF-1α levels by inhibiting VHL and HIF-1α interactions by novel small bioactive molecules, accelerating diabetic wound healing. A three features (two aromatic rings and one hydrogen bond acceptor) pharmacophore hypothesis was generated from the existing HIF-1α modulators. Virtual screening was done based on the generated pharmacophore, and a library consisting of the top 20 out of 3728 compounds was selected using ZINCPharmer. Of the top 20 molecules, the pyrazole moiety was identified as the top "HIT". Five analogues of pyrazole were designed, and Scifinder ascertained the novelty. The designed compounds were synthesized and characterized by IR, Mass, and NMR. Preliminarily, we have carried out a scratch wound assay using 3T3L1 cell lines. All the synthesized compounds showed significant wound healing activity. Further, to validate the in vitro assay, the compound CI, which showed effective in vitro results was used for in vivo study. Using the diabetes mouse model, comprising streptozotocin-induced (STZ) diabetic mice and scratch wound assay, we demonstrated that inhibiting the VHL and HIF-1α connection is a promising strategy for treating diabetic ulcers. Molecules CI and CP were found to have substantial in silico, in vitro, and in vivo outcomes.

Discovery of potential inhibitors for stat3: ligand based 3D pharmacophore, virtual screening, molecular docking, dynamic studies and in vitro evaluation.

A large analysis of the signal transducer and activator of transcription (STAT3) in cancer is currently being carried out. It regulates gene expression, which is required for normal cellular functions such as differentiation, cell growth, proliferation, survival, maturation, and immunity. A ligand-based pharmacophore model was created using 3 D QSAR pharmacophore generation methodology in Discovery studio 4.1 clients to imagine structurally diverse novel chemical entities as STAT3 inhibitors with improved efficacy. Chemical properties of 48 different derivatives were included in the training package. Hypo1 was chosen as the query model for screening 1,45,000 drug-like molecules from the SPECS database, with these molecules subjected to the Lipinski rule of 5, Verber's rule, and SMART filtration. After filtration, the molecule was examined further using molecular docking analysis on the active site of STAT3. The binding interaction(s) and pharmacophore mapping were used to select the 19 possible inhibitory molecules. These 19 hits were then tested for toxicity using the TOPKAT software. In MD simulations and MM-PBSA calculations, the tested compound specs 28 provided the best results, suggesting that this ligand has the ability to inhibit more effectively. Based in-silico finding 19 compounds are subjected to in vitro anticancer activity against MDA-MB-231 and MCF-7 cell lines. Based on results compounds specs 11 and specs 13 shows significant activity compared to other compounds and these compounds were subjected to apoptosis assay. The tested compounds induced morphologic changes were dose and time dependent by which all the tested compound exhibits stronger anti-tumor effects.Communicated by Ramaswamy H. Sarma.