Spiro acenapthene derivatives are structurally diverse group of compounds that show the wide range of antituberculosis activity. The computational evaluation of spiro acenapthene derivatives as rpoB protein from Mycobacterium tuberculosis inhibitor has been performed. The gene rpoB encodes the β–subunit of RNA polymerase. Since no crystal structure was available for the rpoB protein, the three–dimensional structure of rpoB protein from Mycobacterium tuberculosis has been modeled to see the drug interactions,thus allowing us to predict the mutations D435V, H445Y, S450L in the modeled structure. The docking analysis was performed by PyRx software using Autodock 4. The study provides the base for further in vitro and in vivo study of the spiro acenapthene derivatives as rpoB protein inhibitor.