Cynoactanilides 1a, b were reacted with o-hydroxyaldehydes under different conditions. Thus, cyclocondensation of 1a, b with salicylaldehyde, 2-hydroxynaphthal-dehyde and 7-hydroxy-5-methoxy-coumarin-6-carboxaldehyde in ethanolic ammonium acetate afforded the corresponding 2-iminochromene derivatives 3, 7 and 9, respectively. On the other hand, repeating the same reaction in presence of Ac2O/AcONa, the corresponding chromen-2-one derivatives 4, 8 and 10 were obtained. Compounds 4, 8 and 10 were also synthesized through the hydrolysis of 2- imino derivatives using EtOH/HCl. A number of chromeno[3,4-c]pyridine derivatives 11, 13, 14 and 16 were prepared from the reaction of 2-iminochromenes with activated nitriles. Most of the synthesized compounds were screened in vitro for their antibacterial and antifungal activities, by measuring the minimal inhibitory concentration values (MICs), against strains of S. aureus, S. epidermidis , B. subtilis, P. vulgaris, K. pneumonia, S. flexneri, A. fumigates, A.clavatus and Candida albicans using two fold serial dilution method. Ampicillin, gentamycin and amphotricin B were used as reference standards for antibacterial and antifungal activities respectively. Some of the synthesized compounds have been found to exhibit considerable antibacterial and antifungal activities, reaching in certain cases, the same level of antimicrobial activity as the reference standard antibacterial agent Ampicilline and antifungal agent Flucanazole, especially compound 3b that showed promising broad spectrum antibacterial and antifungal properties against all the tested strains. Generation of 3D pharmacophore model was combined to explore the structural requirements controlling the observed antibacterial properties estimated by the effect of the synthesized compounds.