Multiple Myeloma (MM) is highly sensitive to disruptions in cellular protein homeostasis. Proteasome inhibitors (PIs) are initially effective in the treatment of MM, although cures are not achievable and the emergence of resistance limits the durability of responses. New therapies are needed for refractory patients, and those that combat resistance to standard of care agents would be particularly valuable. Screening of multiple chemical libraries for PI re-sensitizing compounds identified E61 as a potent enhancer of multiple PIs and MM specific activity. Using a tandem approach of click chemistry and peptide mass fingerprinting, we identified multiple protein disulfide isomerase (PDI) family members as the primary molecular targets of E61. PDIs mediate oxidative protein folding, and E61 treatment induced robust ER and oxidative stress responses as well as the accumulation of ubiquitinylated proteins. A chemical optimization program led to a new structural class of indene (exemplified by lead E64FC26), which are highly potent pan-style inhibitors of PDIs. In mice with MM, E64FC26 improved survival and enhanced the activity of bortezomib without any adverse effects. This work demonstrates the potential of E64FC26 as an early drug candidate and the strategy of targeting multiple PDI isoforms for the treatment of refractory MM and beyond.
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N.G.D. is an investigator in the South Carolina Center of Biomedical Research Excellence (COBRE) in Oxidants, Redox Balance and Stress Signaling (P20GM103542) and is a Research Scholar of the American Cancer Society (RSG-14-156-01-CDD). This work was also supported by NIH/NCI (1R41CA213488-01), the South Carolina Clinical & Translational Research Institute with an academic home at the Medical University of South Carolina (MUSC; UL1 RR029882 and UL1 TR000062), shared resources of the MUSC Hollings Cancer Center (P30 CA138313), and by the Hollings Cancer Center T32 Ruth L. Kirschstein National Research Service Award Training Program T32 (CA193201). We would also like to thank Pablo Sobrado at Virginia Tech for the generous gifts of the pVP56K vector and the TEV-containing pVP67K vector.
RMR, LR, RMD, HB, MES, and NGD performed research. ABR directed synthesis of E64FC26. JJM and CJC provided support for organic synthesis of E61 derivatives. YM provided support for ROS assays. MMC performed MS of proteins from pull-downs. MC and PLB directed Vk*Myc experiments. RMR and NGD designed research, analyzed data, and wrote the manuscript. All authors reviewed and authorized the final version.
Conflict of interest
RMR and NGD are inventors on patents. NGD has an ownership interest in Leukogene Therapeutics, Inc. The other authors declare that they have no conflict of interest.
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Robinson, R.M., Reyes, L., Duncan, R.M. et al. Inhibitors of the protein disulfide isomerase family for the treatment of multiple myeloma. Leukemia 33, 1011–1022 (2019). https://doi.org/10.1038/s41375-018-0263-1
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