6-Diazo-5-oxo-l-norleucine (DON) is a glutamine antagonist that suppresses cancer cell metabolism but concurrently enhances the metabolic fitness of tumor CD8+ T cells. DON showed promising efficacy in clinical trials; however, its development was halted by dose-limiting gastrointestinal (GI) toxicities. Given its clinical potential, we designed DON peptide prodrugs and found DRP-104 [isopropyl(S)-2-((S)-2-acetamido-3-(1H-indol-3-yl)-propanamido)-6-diazo-5-oxo-hexanoate] that was preferentially bioactivated to DON in tumor while bioinactivated to an inert metabolite in GI tissues. In drug distribution studies, DRP-104 delivered a prodigious 11-fold greater exposure of DON to tumor versus GI tissues. DRP-104 affected multiple metabolic pathways in tumor, including decreased glutamine flux into the TCA cycle. In efficacy studies, both DRP-104 and DON caused complete tumor regression; however, DRP-104 had a markedly improved tolerability profile. DRP-104's effect was CD8+ T cell dependent and resulted in robust immunologic memory. DRP-104 represents a first-in-class prodrug with differential metabolism in target versus toxicity tissue. DRP-104 is now in clinical trials under the FDA Fast Track designation.

Department of Medicine Johns Hopkins School of Medicine Baltimore MD 21205 USA

Department of Neurology Johns Hopkins School of Medicine Baltimore MD 21205 USA

Department of Neuroscience Johns Hopkins School of Medicine Baltimore MD 21205 USA

Department of Oncology Johns Hopkins School of Medicine Baltimore MD 21205 USA

Department of Pharmacology and Molecular Sciences Johns Hopkins School of Medicine Baltimore MD 21205 USA

Department of Psychiatry and Behavioral Sciences Johns Hopkins School of Medicine Baltimore MD 21205 USA

Institute of Organic Chemistry and Biochemistry Academy of Sciences of the Czech Republic v v i Prague 16000 Czech Republic

Johns Hopkins Drug Discovery Johns Hopkins School of Medicine Baltimore MD 21205 USA

The Bloomberg Kimmel Institute for Cancer Immunotherapy Johns Hopkins School of Medicine Baltimore MD 21287 USA

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Pro-905, a Novel Purine Antimetabolite, Combines with Glutamine Amidotransferase Inhibition to Suppress Growth of Malignant Peripheral Nerve Sheath Tumor

Discovery of tert-Butyl Ester Based 6-Diazo-5-oxo-l-norleucine Prodrugs for Enhanced Metabolic Stability and Tumor Delivery