The synthesis of a novel series of 2-mercaptobenzoxazole hydrazide(2-benzo[d]oxazol-2-ylthio) acetohydrazide) derivatives (3a-c, 4a, and 4b) through the use of various  sulfonyl chlorides in dichloromethane  and triethyl amine for the compounds (3a-c) and different aromatic aldehydes in the presence of glacial acetic acid for the compounds (4a, b). The novel compounds' anti-microbial effectiveness has been evaluated in vitro against two of (G+Ve) and two of (G-Ve) bacteria and for fungal activity. In comparison to other compounds, compounds (3a and 3b) had the most anti-microbial action against  Candida albicans and pseudomonas aeruginosa, respectively, while compound (3c)   was more effective against gram-positive bacteria (Streptococcus pyogenes) compare with other compounds. By using their ((melting point,  Thin Layer Chromato-graphy , attenuated total reflectance-Fourier transform infrared  (ATR-FTIR), and ¹ HNMR, the target compounds were identified.