6-Diazo-5-oxo-L-norleucine

6-Diazo-5-oxo-L-norleucine

Systematic (IUPAC) name
(5S)-5-Amino-1-diazonio-6-hydroxy-6-oxohex-1-en-2-olate
Clinical data
Legal status	not approved
Identifiers
CAS Number	157-03-9 Y
PubChem	CID: 5359375
ChemSpider	16735775 N
ChEMBL	CHEMBL97485 N
Chemical data
Formula	C6H9N3O3
Molecular mass	171.15 g/mol
SMILES O=C(CC[C@H](N)C(O)=O)\C=[N+]=[N-]
InChI InChI=1S/C6H9N3O3/c7-5(6(11)12)2-1-4(10)3-9-8/h3,5H,1-2,7H2,(H,11,12)/t5-/m0/s1 N Key:YCWQAMGASJSUIP-YFKPBYRVSA-N N
NY (what is this?)  (verify)

6-Diazo-5-oxo-L-norleucine (DON) is a glutamine antagonist, which was isolated originally from Streptomyces in a sample of Peruvian soil. It is a non-standard amino acid. The diazo compound 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. The last clinical results were published in 2008, though not as DON monotherapy but in combination with a recombinant glutaminase.

Chemistry

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]₂₆^D +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).

Biochemistry

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. In the following table gives a survey of DON targets.

Selection of enzymes inhibited by DON

Enzyme	Metabolic pathway	References
Carbamoyl phosphate synthase (CAD)	Pyrimidine-De-Novo-Synthesis
CTP synthase (CTPS)	Pyrimidine-De-Novo-Synthesis
FGAR amidotransferase	Purine-De-Novo-Synthesis
Guanosine monophosphate synthetase (GMPS)	Purine-De-Novo-Synthesis
PRPP amidotransferase	Purine-De-Novo-Synthesis
Mitochondrial glutaminase	First step of glutaminolysis
NAD synthase	Coenzyme of the electron transport chain
Asparagine synthetase	Amino acid synthesis

Pharmacology

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.