Nigella sativa is considered a miracle herb plant belongs to the Ranunculaceae family, inhabitant to South Asia and Southwest Asia. The most important part of the Nigella sativa plant used by human beings are seeds, mostly as a spice because it contains a specific aroma. The seeds derived from Nigella sativa being exploited by people and used for thousands of years for culinary purposes, to cure various digestive disorders, recover from joint pains, obesity, common cold, headache, asthma, rheumatic diseases, etc. in India, Pakistan, Bangladesh, China, and Middle East countries. In the current study, the seed extracts of Nigella sativa were targeted for evaluation of qualitative biochemical analysis of phytochemicals, antibacterial, cytotoxicity, and antioxidant properties. Nanoparticles formulations derived as of the aqueous extract of seeds of Nigella sativa were developed and tested against selected microorganisms. FTIR and HPLC also performed to evaluate the functional group analysis and to detect the presence of a bioactive compound in the phytochemical extract. Cytotoxicity properties of methanolic and ethanolic seed extracts of Nigella sativa have been evaluated against three cell lines; A549, SW480, and HeLa. In the current findings, bioactive constituents present in extracts were alkaloids, flavonoids, saponins, tannins, and phenolic compounds which are responsible for antioxidant activities, antibacterial, and cytotoxic effects. N. sativa seeds contain high valuable bioactive molecules which can perform a very significant role in curing various diseases and in development of the green drug and can be explored for immunomodulatory effects. INTRODUCTION: Nigella sativa is mostly named as black seed or black cumin or kalonji seeds, and in Middle East countries, it is traditionally called as "Haba al-barakah" meaning seeds of blessings, belongs to the plant family Ranunculaceae.