Echinomycin,　a　member　of　the　quinoxaline　family　of　antibiotics,　is　known　to　be　a　small-molecule　inhibitor　of　hypoxia　inducible　factor-1　(HIF-1)　DNA　binding　activity.　Recently,　it　has　been　shown　to　suppress　mammalian　target　of　rapamycin　(mTOR)　signaling　and　growth　in　leukemia　cell　lines.　In　this　study,　we　investigated　whether　echinomycin　interacts　with　the　FKBP12　protein.　Molecular　docking　was　used,　and　the　predicted　binding　energy　was　-10.61　kcal/mol.　Moreover,　surface　plasmon　resonance　imaging　and　fluorescence　quenching　techniques　were　used　to　validate　this　interaction.　Echinomycin　binds　to　FKBP12　with　a　strong　binding　affinity　comparable　with　rapamycin.　Furthermore,　the　echinomycin-FKBP12　complex　has　been　shown　to　affect　calcineurin　activity　when　tested　in　a　calcineurin　phosphatase　inhibition　assay.　All　of　these　studies　have　shown　that　echinomycin　may　have　a　double　impact　on　HIF　signaling　by　direct　inhibition　and　through　mTOR.