AbstractThe fungal natural product aspergillomarasmine A (AMA) has been identified as a noncompetitive inhibitor of New Delhi metallo‐β‐lactamase‐1 (NDM‐1) that inhibits by removing ZnII from the active‐site. The nonselective metal‐chelating properties and difficult synthesis and derivatization of AMA have hindered the development of this scaffold into a potent and selective inhibitor of NDM‐1. Iminodiacetic acid (IDA) has been identified as the metal‐binding pharmacophore (MBP) core of AMA that can be leveraged for inhibitor development. Herein, we report the use of IDA for fragment‐based drug discovery (FBDD) of NDM‐1 inhibitors. IDA (IC50=120 μM) was developed into inhibitor 23 f (IC50=8.6 μM, Ki=2.6 μM), which formed a ternary complex with NDM‐1, as evidenced by protein thermal‐shift and native‐state electrospray ionization mass spectrometry (ESI‐MS) experiments. Combining mechanistic analysis with inhibitor derivatization, the use of IDA as an alternative AMA scaffold for NDM‐1 inhibitor development is detailed.