The DevR (DosR) response regulator is central to hypoxia-induced dormant state associated with drug tolerance in Mycobacterium tuberculosis. DevR can be activated independently of its sensor kinases via acetyl phosphate, underscoring the importance of directly targeting DevR, thereby establishing it as an attractive therapeutic target. Here, we report the design, synthesis, characterization and evaluation of a novel series of Coumarin-Phenyl Urea (CPU1–CPU21) derivatives targeting DevR. At 250 μM, four compounds (CPU3, CPU7, CPU9, and CPU19) significantly inhibited in vitro DNA binding activity of DevR. Molecular docking identified key DevR binding residues, with CPU3 showing the most favorable scores. Thermal shift assays confirmed their direct interaction with DevR. CPU3 demonstrated 55% inhibition of mycobacterial growth when added initially and post hypoxia establishment. Mechanistically, CPU3 disrupted in vitro dimerization of DevR. Thus, CPU3 is identified as a promising DevR-targeting scaffold with a dual mechanism, offering potential for therapeutic strategies against latent tuberculosis.
Pancreatic ductal adenocarcinoma (PDAC) is an aggressive malignancy, rapidly progresses and exhibits metabolic reprogramming and resistance to conventional therapies. A systems biology and computational integrative approach used in this study to explore important metabolic enzymes, such as ME1, NAPRT, NQO1 and PECR, and to assess the multi-target inhibitory ability of oxysanguinarine (oSNG). Overexpression and functional crosstalk of these genes leading to the establishment of a coordinated gene network involved in the biosynthesis of NAD+, NADPH, oxidative stress adaption and lipid metabolism. Physicochemical & pharmacokinetic analysis and DFT study revealed that oSNG has drug likeness, stability and chemical reactivity. All the targets namely ME1, NAPRT, NQO1 and PECR has high affinity for the oSNG (-9.8, -8.2, -10.2 and -8.8 kcal/mol respectively). Molecular dynamic simulations validate the stability and conformational integrity of complexes. oSNG shows potent cytotoxicity with IC₅₀ = 11.7 ± 2.69 μM (48 h) and inhibition of cell migration in PANC-1 cells. Its cytotoxic effects are associated with elevated intracellular ROS levels, leading to oxidative stress and mitochondrial membrane potential (ΔΨm) disruption. These findings suggest therapeutic potential of oSNG mediated by ROS and mitochondrial dysfunction.
Declining sperm counts are increasingly reported worldwide, yet their biological and clinical significance remains uncertain. Although meta-analyses suggest a long-term downward trend, interpretation is limited by methodological variation and an overreliance on sperm count as a single marker of male reproductive health. This creates a critical gap between population level observations and underlying biology, leaving many cases labelled as idiopathic despite measurable dysfunction. This review addresses this gap by proposing a hypothesis driven framework that integrates oxidative stress, inflammation, and metabolic dysfunction as interacting drivers of male infertility. Rather than acting independently, environmental exposures, lifestyle factors, and developmental influences converge on shared biological pathways that impair spermatogenesis and sperm function. This model explains the variability in clinical presentation while accounting for consistent population level decline. By shifting the focus from isolated semen parameters to interconnected biological processes, the framework provides a basis for mechanism-based classification and targeted management. It also supports a more coherent interpretation of epidemiological trends. Declining sperm counts are therefore better understood not as an isolated finding, but as an indicator of broader systemic dysfunction with implications for reproductive and general health.
The development of advanced cell culture models has overcome the limitations of conventional monolayer cultures, still doubts remain about the reliability of data obtained using traditional systems, as well as the comparability of results from different models. This is highly relevant to preclinical drug analysis, where in vitro studies determine the fate of molecules. We investigated the molecular mechanisms that regulate the activity of cannabidiolic acid (CBDA) in a 3D model of a glioblastoma cell line, comparing the results with those obtained on a conventional monolayer model of the same cells. CBDA targeted the translation initiation factor EIF2A in both tested models. However, the downstream consequences of the CBDA-EIF2A interaction differed between cells cultured in 3D and 2D, since the biological functions and interactome of EIF2A were found to change significantly. Overall, this study sheds new light on the difficulties associated with comparing results obtained using different in vitro models.
A novel series of pyrazolo[3,4-d]pyrimidine derivatives conjugated with uracil, cytosine, and benzene sulfonamide moieties (5a-11b) were designed as dual inhibitors of tumor-associated carbonic anhydrase (CA) isoforms and and 11b emerged as the most potent candidates, exhibiting nanomolar inhibition against CA II, IX, and XII. Notably, 11a inhibited CA IX with a Ki of 2.6 nM, while 11b showed potent inhibition of CA II (Ki= 7.1 nM) and CA IX (Ki = 2.8 nM). Furthermore, both compounds surpassed erlotinib in EGFR inhibition, with IC50 values of 69 nM (11a) and 64 nM (11b). Biological evaluation revealed significant antiproliferative activity against MCF-7, HePG-2, HCT-116, and A549 cancer cell lines under normoxic and hypoxic conditions. Under hypoxia, 11a demonstrated superior potency (IC50 range: 4.04-9.58 & micro;M). Flow cytometry indicated that 11a induced G1 phase arrest, while 11b triggered G2/M arrest and pronounced apoptosis, reducing cell viability to 12.4%. Molecular analysis confirmed the activation of the intrinsic apoptotic pathway through the upregulation of BAX, caspase-9, and p53, alongside Bcl-2 downregulation. Molecular docking and 100 ns molecular dynamics simulations confirmed stable binding within CA-II, CA-IX, and EGFR active sites. These findings identify 11b as a highly promising multi-target lead for further anticancer development.
