Steroidal Pseudo‐Alkaloids via 2‐Cyanoacetohydrazide Post‐Functionalization: Synthesis, Biological Profiling, and Computational Insights

ABSTRACT Steroids constitute distinct bioactive scaffolds whose rigidity and structural complexity hinder the development of novel hybrid derivatives. To address this, methyltestosterone ( MT ) was treated with 2‐cyanoacetohydrazide to afford a versatile precursor ( 1 ) for synthesizing diverse steroidal pseudo‐alkaloids. This approach yielded uracil ( 2a ), thiouracil ( 2b ), and azine ( 4) derivatives, while further transformations produced hydrazide–hydrazone ( 5 ), 2,6‐dioxo‐tetrahydropyridine ( 6 ), and dihydro‐6 H ‐1,3‐oxazin‐6‐one ( 8 ). Biological evaluation identified compound 8 as the most active derivative, showing promising cytotoxicity against HepG‐2 and MCF‐7 cells (IC 50 = 10.23 ± 0.8 and 6.90 ± 0.3 µM, respectively) and antimicrobial activity against E. coli (MIC = 8.0 µM), S. aureus (MIC = 4.0 µM), B. subtilis (MIC = 16.0 µM), and C. albicans (MIC = 8.0 µM). Molecular docking revealed that compound 8 interacted mainly via hydrophobic contacts with VEGFR2, YcbB, and CYP51, whereas compound 1 exhibited the strongest binding to the human androgen receptor (hAR‐LBD; docking score = −13.19 kcal/mol). DFT calculations at the B3LYP/6‐31G(d,p) level, in addition to wavefunction analysis, further showed that pseudo‐alkaloidization of MT modulated the steroidal framework's electronic properties and polarity, with compound 8 exhibiting a relatively narrow energy gap of 3.99 eV and a dipole moment of 4.12 D, consistent with its biological activity.