This study examines the design and synthesis of a newly manufactured pyrrolo[2,3-d]pyrimidinehydrazide (PPH) derivative. The PPH derivatives are designed to contain bromine atoms in positions 2, 3, 4, 5, and 6. The microwave technique is used as a novel and reliable method for the preparation of this type of PPH derivative. Spectral and elemental studies were used to provide a comprehensive description of the chemical structure of the PPH derivatives 2–7 that were constructed. By using the disc diffusion method, each drug was put through an in vitro antimicrobial test against six different microorganisms. In comparison to streptomycin, which had a concentration of 25.0 μg/ml, it was discovered that compounds 2, 4, and 5 exhibited the highest levels of activity against E. coli (11.5, 15.5, and 23.4 μg/ml, respectively). Furthermore, compound 7 demonstrated potential cytotoxic effects against artemia salina, with an IC50 value of 18.5 μg/ml.