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Mammary gland carcinoma is one of the most prevalent and deadly diseases among women globally. It is a type of solid malignant tumor. In this malignant tumor, the microenvironment becomes hypoxic in rapidly proliferating cancer cells. These cells undergo adaptive changes through the expression of hypoxia‐inducible factor‐1alpha (HIF‐1α) which is regulated by factor inhibiting HIF‐1α (FIH‐1). Considering this, we hypothesized that the chemical activation of FIH‐1 would inhibit the hypoxic activity of HIF‐1α in mammary gland carcinoma. A library of 67,609 chemical compounds was virtually screened against FIH‐1 based on Lipinski's rule from the ZINC database. The BBAP‐8 has been selected based on an excellent docking score (−8.352 Kcal/mol), favorable ADMET, and potential FIH‐1 activator profile. Further, its in‐vitro cytotoxicity and apoptotic activity were scrutinized against MCF‐7 cells and in‐vivo activity against 7,12‐dimethylbenz[a]anthracene (DMBA) induced mammary gland carcinoma in Wistar rats. It exhibited significant cytotoxicity (IC50 = 16.59 ± 0.49 μM) and activated apoptosis when scrutinized through DAPI, AO/EB, and JC‐1 staining. Also, oral administration of BBAP‐8 restored hemodynamic changes, normalized tissue architecture, and corrected metabolic abnormalities. The western blot analysis and mRNA expression analysis validated that BBAP‐8 has the potential to activate FIH‐1 with the downregulation of GLUT‐1, VEGF, and Twist‐1. Moreover, BBAP‐8 fostered apoptosis, when evaluated through BCL‐2, BAX, Caspase‐8, and Caspase‐3. Based on research findings, this implies that BBAP‐8 activates FIH‐1 and can be effective in chemotherapeutic treatment of mammary gland carcinoma. [ABSTRACT FROM AUTHOR]

Inhibition of prolylhydroxylase‐2 (PHD‐2) in both normoxic and hypoxic cells is a critical component of solid tumours. The present study aimed to identify small molecules with PHD‐2 activation potential. Virtually screening 4342 chemical compounds for structural similarity to R59949 and docking with PHD‐2. To find the best drug candidate, hits were assessed for drug likeliness, antihypoxic and antineoplastic potential. The selected drug candidate's PHD‐2 activation, cytotoxic and apoptotic potentials were assessed using 2‐oxoglutarate, MTT, AO/EtBr and JC‐1 staining. The drug candidate was also tested for its in‐vivo chemopreventive efficacy against DMBA‐induced mammary gland cancer alone and in combination with Tirapazamine (TPZ). Virtual screening and 2‐oxoglutarate assay showed BBAP‐6 as lead compound. BBAP‐6 exhibited cytotoxic and apoptotic activity against ER+ MCF‐7. In carmine staining and histology, BBAP‐6 alone or in combination with TPZ restored normal surface morphology of the mammary gland after DMBA produced malignant alterations. Immunoblotting revealed that BBAP‐6 reduced NF‐κB expression, activated PHD‐2 and induced intrinsic apoptotic pathway. Serum metabolomics conducted with 1H NMR confirmed that BBAP‐6 prevented HIF‐1α and NF‐κB‐induced metabolic changes in DMBA mammary gland cancer model. In a nutshell, it can be concluded that BBAP‐6 activates PHD‐2 and exhibits anticancer potential. [ABSTRACT FROM AUTHOR]

Normoxic inactivation of prolyl hydroxylase-2 (PHD-2) in tumour microenvironment paves the way for cancer cells to thrive under the influence of HIF-1a 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. [ABSTRACT FROM AUTHOR]