Affiliation:
1. Department of Pharmaceutical Sciences, School of Biomedical and Pharmaceutical Sciences Babasaheb Bhimrao Ambedkar University (A Central University) Lucknow Uttar Pradesh India
2. Department of Pharmacology, College of Pharmacy Prince Sattam Bin Abdulaziz University Al‐Kharaj Saudi Arabia
3. Department of Pharmaceutical Sciences, United Institute of Pharmacy United Group of Institutions Prayagraj India
Abstract
AbstractNormoxic inactivation of prolyl hydroxylase‐2 (PHD‐2) in tumour microenvironment paves the way for cancer cells to thrive under the influence of HIF‐1α and NF‐κB. Henceforth, the present study is aimed to identify small molecule activators of PHD‐2. A virtual screening was conducted on a library consisting of 265,242 chemical compounds, with the objective of identifying molecules that exhibit structural similarities to the furan chalcone scaffold. Further, PHD‐2 activation potential of screened compound was determined using in vitro 2‐oxoglutarate assay. The cytotoxic activity and apoptotic potential of screened compound was determined using various staining techniques, including 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl tetrazolium bromide, 4′,6‐diamidino‐2‐phenylindole (DAPI), 1,1′,3,3′‐tetraethylbenzimi‐dazolylcarbocyanine iodide (JC‐1), and acridine orange/ethidium bromide (AO/EB), against MCF‐7 cells. 7,12‐Dimethylbenz[a]anthracene (DMBA) model of mammary gland cancer was used to study the in vivo antineoplastic efficacy of screened compound. [(E)‐1‐(4‐fluorophenyl)‐3‐(furan‐2‐yl) prop‐2‐en‐1‐one] (BBAP‐7) was screened and validated as a PHD‐2 activator by an in vitro 2‐oxo‐glutarate assay. The IC50 of BBAP‐7 on MCF‐7 cells is 18.84 µM. AO/EB and DAPI staining showed nuclear fragmentation, blebbing and condensation in MCF‐7 cells following BBAP‐7 treatment. The red‐to‐green intensity ratio of JC‐1 stained MCF‐7 cells decreased after BBAP‐7 treatment, indicating mitochondrial‐mediated apoptosis. DMBA caused mammary gland dysplasia, duct hyperplasia and ductal carcinoma in situ. Carmine staining, histopathology, and scanning electron microscopy demonstrated that BBAP‐7, alone or with tirapazamine, restored mammary gland surface morphology and structural integrity. Additionally, BBAP‐7 therapy significantly reduced oxidative stress and glycolysis. The findings reveal that BBAP‐7 activates PHD‐2, making it a promising anticancer drug.