Herein, a fast and convenient protocol for the synthesis of new isoniazid fused chromeno[4,3-b]quinolin was achieved through biomimetic catalysis by cyclodextrin in the water at 60–65 °C. The present investigation involves attractive characteristics such as the use of water as the reaction medium, one-pot conditions, short reaction periods, easy work-up/purification and reduced waste production. This method provides a green route for the synthesis of targeted scaffolds and also a wide substrate scope for several substituted aldehydes to provide good yields of the corresponding products. Furthermore, the catalyst can be easily recovered by simple filtration and reused several times without any substantial loss in activity. Our study also discloses the antimicrobial screening of new chromeno[4,3-b]quinolin-isonicotinamides against four bacterial and three fungal strains.