|Chemical and physical data|
|Molar mass||171.156 g·mol−1|
|3D model (JSmol)|
|(what is this?)|
6-Diazo-5-oxo-L-norleucine (DON) is a glutamine antagonist, which was isolated originally from Streptomyces in a sample of Peruvian soil. This diazo compound is biosynthesized from lysine by three enzymes in bacteria. It is one of the most famous non-proteinogenic amino acid and was characterized in 1956 by Henry W Dion et al., who suggested a possible use in cancer therapy. This antitumoral efficacy was confirmed in different animal models. DON was tested as chemotherapeutic agent in different clinical studies, but was never approved. In 2019, DON was shown to kill tumor cells while reversing disease symptoms and improve overall survival in late-stage experimental glioblastoma in mice, when combined with calorie-restricted ketogenic diet.
DON is a water-soluble yellowish powder, which can be dissolved also in aqueous solutions of methanol, acetone or ethanol, but dissolution in absolute alcohols is difficult. Solutions of at least 50 μM DON in 0.9% NaCl are lightly yellowish. The crystalline form appears as yellowish greenish needles. The specific rotation is [α]26D +21° (c = 5.4% in H2O). In phosphate buffer, pH 7 are the ultraviolet absorption maxima at 274 nm (E1%1 cm. 683) and 244 nm (E1%1 cm 376).
DON is used as inhibitor of different glutamine utilizing enzymes. Due to its similarity to glutamine it can enter catalytic centres of these enzymes and inhibits them by covalent binding, or more precisely by alkylation. The following table gives a survey of DON targets.
|Carbamoyl phosphate synthase (CAD)||Pyrimidine-De-Novo-Synthesis|||
|CTP synthase (CTPS)||Pyrimidine-De-Novo-Synthesis|||
|Guanosine monophosphate synthetase (GMPS)||Purine-De-Novo-Synthesis|||
|Mitochondrial glutaminase||First step of glutaminolysis|||
|NAD synthase||Coenzyme of the electron transport chain|||
|Asparagine synthetase||Amino acid synthesis|||
DON is a cytotoxic inhibitor of many enzymes of nucleotide synthesis. It could be shown in vitro that DON treatment led to apoptosis, the programmed cell death. Different pathways were investigated. So it could be shown that the inner mitochondrial membrane was damaged, and single strand DNA breaks were observed. The exact mode of action remains unclear and needs further research.
DON is not approved as pharmaceutical agent, but is tested in combination with a recombinant glutaminase in clinical trials for the treatment of different solid tumors.
- JHU-083, a prodrug of 6-diazo-5-oxo-L-norleucine
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- Mukherjee P, Augur ZM, Li M, Hill C, Greenwood B, Domin MA, et al. (29 May 2019). "Therapeutic benefit of combining calorie-restricted ketogenic diet and glutamine targeting in late-stage experimental glioblastoma". Communications Biology. 2 (1): 200. doi:10.1038/s42003-019-0455-x. PMC 6541653. PMID 31149644.
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- Rosenbluth RJ, Cooney DA, Jayaram HN, Milman HA, Homan ER (August 1976). "DON, CONV and DONV-II. Inhibition of L-'asparagine synthetase in vivo". Biochemical Pharmacology. 25 (16): 1851–1858. doi:10.1016/0006-2952(76)90189-1. PMID 9091.
- Wu F, Lukinius A, Bergström M, Eriksson B, Watanabe Y, Långström B (July 1999). "A mechanism behind the antitumour effect of 6-diazo-5-oxo-L-norleucine (DON): disruption of mitochondria". European Journal of Cancer. 35 (7): 1155–1161. doi:10.1016/S0959-8049(99)00099-4. PMID 10533463.
- Hiramoto K, Fujino T, Kikugawa K (June 1996). "DNA strand cleavage by tumor-inhibiting antibiotic 6-diazo-5-oxo-L-norleucine". Mutation Research. 360 (2): 95–100. doi:10.1016/0165-1161(95)00073-9. PMID 8649470.
- Mueller C, Al-Batran S, Jaeger E, Schmidt B, Bausch M, Unger C, Sethuraman N (2008). "A phase IIa study of PEGylated glutaminase (PEG-PGA) plus 6-diazo-5-oxo-L-norleucine (DON) in patients with advanced refractory solid tumors". J Clin Oncol. 26 (May 20 Suppl): abstr 2533. doi:10.1200/jco.2008.26.15_suppl.2533.