A series of fourteen chalcone was synthesized via. Claisen–Schmidt condensation between substituted 2- hydroxyl acetonaphthones and substituted benzaldehyde in presence of tripotassium phosphate (K3PO4) catalyst. The reaction was carried out by conventional method using 2-methoxyethanol. The procedure is simple and efficient in terms of reaction time, easy workup and isolation of products and yields. In-vitro all these synthesized compounds were screened and evaluated for the cytotoxic and antimicrobial activity. It was found that these compounds had significant cytotoxic activity in comparison with standard 5-flurouracil. The compounds 3a, 3b, 3h, 3f and 3l were screened by MTT assay against liver cancer cell line-HepG2. Among these, the compound 3b and 3c showed LC50 values of 997.14 μM/ml and 284.13 μM/ml., respectively. The remaining compounds did not display the LC50 values. The compound 3l displayed the strongest cytotoxic activities with IC50 value of 91.85 μg/ml against liver cancer cell line. The Chalcone 3a, 3f, 3h and 3e demonstrated excellent antimicrobial activity and the remaining were moderately active against tested pathogens. The antimicrobial effects of all the tested compounds are due to the presence of pharmacological active substituent in the basic nucleus of Chalcones. Therefore, the present study leads to the development of new class of anticancer and antimicrobial inhibitory candidates.