The Macrophage infectivity potentiator (Mip) protein is a virulence factor encoded by intracellular pathogens. Burkholderia pseudomallei, the causative agent of melioidosis is a disease endemic in South East Asia and Northern Australia. There is currently no vaccine available against B. pseudomallei and the organism is resistant to a range of antibiotics, therefore identification of new drug targets is essential. The Mip protein in B. pseudomallei exhibits virulence-associated peptidyl-prolyl isomerase (PPIase) activity, inhibition of which potentially represents a novel target for antimicrobial therapies to this organism.. A group of pipecolic acid derivatives have been shown to inhibit Mip from L. pneumophila. Using a protease coupled PPIase assay, we have demonstrated that a selection of these pipecolic acid derived compounds also have inhibitory properties against the recombinant B. pseudomallei Mip. Importantly, these inhibitors reduce the cytotoxic effects of B. pseudomallei on macrophages. Furthermore we have demonstrated that these inhibitors have broad-spectrum activity against Gram-negative bacteria.