Bis-acyl-thiourea derivatives, namely N,N’-(((4-nitro-1,2-phenylene)bis(azanediyl)) bis(carbonothioyl))bis(2,4-dichlorobenzamide) (UP-1), N,N’-(((4-nitro-1,2-phenylene) bis(azanediyl))bis(carbonothioyl))diheptanamide (UP-2), and N,N’-(((4-nitro-1,2-phenylene)bis(azanediyl))bis(carbonothioyl))dibutannamide (UP-3), were synthesized in two steps. The structural characterization of the derivatives was carried out by FTIR, ¹H-NMR, and ¹³C-NMR, and then their DNA binding, anti-urease, and anticancer activities were explored. Both theoretical and experimental results, as obtained by density functional theory, molecular docking, UV-visible spectroscopy, fluorescence (Flu-)spectroscopy, cyclic voltammetry (CV), and viscometry, pointed towards compounds’ interactions with DNA. However, the values of binding constant (K_b), binding site size (n), and negative Gibbs free energy change (ΔG) (as evaluated by docking, UV-vis, Flu-, and CV) indicated that all the derivatives exhibited binding interactions with the DNA in the order UP-3 > UP-2 > UP-1. The experimental findings from spectral and electrochemical analysis complemented each other and supported the theoretical analysis. The lower diffusion coefficient (D_o) values, as obtained from CV responses of each compound after DNA addition at various scan rates, further confirmed the formation of a bulky compound–DNA complex that caused slow diffusion. The mixed binding mode of interaction as seen in docking was further verified by changes in DNA viscosity with varying compound concentrations. All compounds showed strong anti-urease activity, whereas UP-1 was found to have comparatively better inhibitory efficiency, with an IC₅₀ value of 1.55 ± 0.0288 µM. The dose-dependent cytotoxicity of the synthesized derivatives against glioblastoma MG-U87 cells (a human brain cancer cell line) followed by HEK-293 cells (a normal human embryonic kidney cell line) indicated that UP-1 and UP-3 have greater cytotoxicity against both cancerous and healthy cell lines at 400 µM. However, dose-dependent responses of UP-2 showed cytotoxicity against cancerous cells, while it showed no cytotoxicity on the healthy cell line at a low concentration range of 40–120 µM.

In the current study, Azo-Thiohydantoins derivatives were synthesized and characterized by using various spectroscopic techniques including FTIR, ¹H-NMR, ¹³C-NMR, elemental and HRMS analysis. The compounds were evaluated for alkaline phosphatase activity and it was observed that among all the synthesized compounds, derivative 7e exhibited substantial inhibitory activity (IC₅₀ = 0.308 ± 0.065 µM), surpassing the standard inhibitor (L–Phenyl alanine, IC₅₀ = 80.2 ± 1.1 µM). Along with this, these derivatives were comprehensively examined regarding the electronic properties and reactivity of the synthesized compounds using Density Functional Theory (DFT) calculations, where the results were found very promising and the synthesized compound were found stable. After that, SwissADME evaluations highlighted compounds for their favorable physicochemical properties, including solubility and drug-likeness. Molecular docking exhibited the strong binding affinities of 7f and 7e derivatives with intestinal alkaline phosphatase (IAP), further supported by Molecular Dynamics (MD) simulations. This comprehensive integration of experimental and computational approaches sheds the light on the potential therapeutic applications of the synthesized compounds. By providing a detailed investigation of these aspects, this research opens the avenues for the development of novel pharmacologically active compounds with diverse applications.

In the modern age of drug discovery sulfanilamide derivatives are known to have great anti-cancerous potential, the current study aimed to synthesize these derivatives in order to evaluate their biological properties against carbonic anhydrase II (CA-II) and Dickkopf − 1(Dkk1) protein which are highly expressed in many cancers including lung cancer. A series of 10 sulfanilamide derivatives was synthesized under controlled conditions using reflux condensation method. Among all the synthesized derivatives (5a-5j), the compound 5d was found to possess highest antioxidant activity (90.7397 ± 0.0732 µg/mL) comparable to vitamin C (95.1571 ± 0.057 µg/mL) and also exhibited maximum inhibition against CA-II with an IC₅₀ value of 0.00690 ± 0.1119 µM, indicating that 5d is significantly more potent as compared to standard i.e., acetazolamide IC₅₀ = 0.9979 ± 0.0024 µM. Keeping in view the importance of Dkk1 protein in cancer progression, the molecular docking investigations were performed, where compound 5d was proved to be the potential dual inhibitor of CA-II as well as Dkk1 with the binding energy of 8.9 and 9.7 kcal/mol, respectively. In addition to this DNA binding studies also confirmed the significance of compound 5d where it had maximum binding constant value of 6.7 × 10⁴ mol^− 1, supporting the other biological investigations and was in agreement with the reported values. All the experimental and computational results reveals the excellent potential of 5d as a candidate medicine in future. Conclusively, the current study may lead to the new therapeutic strategies for the treatment of cancer associated with the aberrant expression of CA-II and less explored DDK1 target.

Human Carbonic Anhydrase inhibitors (CAIs) have been clinically used to treat a variety of disorders, such as cancer, obesity, haemolytic anaemia, glaucoma, retinopathy, and epilepsy. To develop a Carbonic Anhydrase inhibitor, Iminothiazoline analogue ((Z)-N-(3-([1,1'-biphenyl]-2-yl)-4-heptyl-4-hydroxythiazolidin-2-ylidene)-4-bromobenzamide) was synthesized and characterized. Single crystal X-Ray diffraction studies and Hirshfeld surface analysis (HSA) were conducted to find the exact molecular structure as well as intermolecular interactions. DFT Calculations indicated the soft and reactive nature of molecule. In-Vitro carbonic anhydrase inhibition studies showed the excellent inhibition potential of (Z)-N-(3-([1,1'-biphenyl]-2-yl)-4-heptyl-4-hydroxythiazolidin-2-ylidene)-4-bromobenzamide (IC50 value of 0.147 ± 0.03 µM). Four hydrogen bonds and a multiple hydrophobic interactions were observed between synthesized molecule and the enzyme during Molecular docking studies. Molecular dynamic simulation studies showed that Protein–ligand complex generally remained stable throughout the time. ADMET studies suggested the need of structural modification for the drug like behavior of synthesized molecule.

Naphthyl Thiourea based derivative N-(ethylcaramothbioyl)-1-naphthamide (NA-MT) was synthesized by freshly prepared 1-naphthoyl isothiocyanate with ethyl amine to afford the products (NA-MT) high purity and characterized via spectroscopic techniques including FTIR, 1H-NMR, 13C-NMR, elemental and HRMS analysis and single crystal X-ray diffraction. In-vitro analysis showed that the compound (NA-MT) possesses potent inhibitory effect with IC50 = 9.875 ± 0.05 surpassing its reference inhibitor L-phenyl alanine (IC50 = 80.2 ± 1.1) against cIAP. Additionally, the synthesized derivative (NA-MT) underwent an in-depth analysis of its electronic properties and reactivity using Density Functional Theory (DFT) calculations. Evaluations using SwissADME revealed the compound (NA-MT) possess acceptable physicochemical attributes, such as solubility and drug-likeness. Molecular docking studies demonstrated the compound (NA-MT) exhibit strong binding affinities to cIAP, which were further validated via Molecular Dynamics (MD) simulations. These integrated experimental and computational tools highlight the potential therapeutic uses of the synthesized compound, and pave the way for the development of novel pharmacologically active Alkaline phosphatase inhibitors with diverse applications.