Objective: To deal with the anti-uropathogenic and in silico screening of(E-)-N’-(substitutedbenzylidene)-2-(quinolin-8-yloxy)acetohydrazide analogues in order to search the potential anti-uropathogenic agents.Methods: Three(E-)-N’-(substituted-benzylidene)-2-(quinolin-8-yloxy)acetohydrazide analogues were synthesized. Structure elucidation was done using various spectroscopic techniques including infrared radiation, 1hydrogen-nuclear magnetic resonance, carbon-13 nuclear magnetic resonance, etc. Physicochemical score, bioactivity score and molecular docking studies were carried out using Lipinski’s rule of five, Molinspiration(web based software), Autodock 4.2 tools. In vitro anti-uropathogenic activity was carried out against four pathogens named as Staphylococcus aureus(S. aureus), Staphylococcus epidermidis, Proteus mirabilis and Escherichia coli by disc diffusion method and macro-dilution test following their morphological and biochemical characterization.Results: The formation of(E-)-N’-(substituted-benz ylidene)-2-(quinolin-8-yloxy)acetohydrazide is confirmed from the spectroscopic results. All the compounds were found in compliance with Lipinski’s rule of five and exhibited bioactivity score from-0.50 to 0.00. Docking results revealed that compound-1 is forming one hydrogen bond with TYR 576 and two hydrogen bond with GLU 569, while compound-2 is forming one hydrogen bond with ARG 599, and compound-3 forming 0 hydrogen bond. The anti-uropathogenic evaluation exhibited that compound one exhibited better activity against S. aureus, while it was found to possess moderate to good activity against both Gram-positive bacteria and Gram-negative bacteria excluding S. aureus.Conclusions: Our study revealed that compound one exhibited better activity than the standard in case of S. aureus and moderate to good activity against rest of the pathogens. Molecular docking, physicochemical and bioactivity studies strongly supported the experimental results. From the well obtained results it was concluded that compound-1 can lead as potential antiuropathogenic agents. Objective: To deal with the anti-uropathogenic and in silico screening of (E -) - N ’- (substitutedbenzylidene) -2- (quinolin-8-yloxy) acetohydrazide analogues in order to search the potential anti-uropathogenic agents. Methods: Structure elucidation was done using various spectroscopic techniques including infrared radiation, 1 hydrogen-nuclear magnetic resonance, carbon-13 nuclear magnetic resonance, etc. Physicochemical score, bioactivity score and molecular docking studies were carried out using Lipinski’s rule of five, Molinspiration (web based software), Autodock 4.2 tools. In vitro anti-uropathogenic activity was carried out against four pathogens named as Staphylococcus aureus (S. aureus), Staphylococcus epidermidis, Proteus mirabilis and Escherichia coli by disc diffusion method and macro-dilution test following their morphological and biochemical characterization. Results: The formation of All (substituted-benz ylidene) -2- (quinolin-8-yloxy) acetohydrazide is confirmed from the spectroscopic results. All the compounds were found in compliance with Lipinski’s rule of five and indicated bioactivity score from- 0.50 to 0.00. Docking results revealed that compound-1 is one hydrogen bond with TYR 576 and two hydrogen bonds with GLU 569, while compound-2 is forming one hydrogen bond with ARG 599, and compound-3 forming 0 hydrogen bond. anti-uropathogenic evaluation phenol that compound one suggesting better activity against S. aureus, while it was found to possess moderate to good activity against both Gram-positive bacteria and Gram-negative bacteria excluding S. aureus. Confclusions: Our study revealed that compound one Homozygotes and activity to the standard in case of S. aureus and moderate to good activity against rest of the pathogens. Molecular docking, physicochemical and bioactivity studies strongly supported the experimental results.ained results it was concluded that compound-1 can lead as potential antiuropathogenic agents.