Nitrosation of Glycine Ethyl Ester and Ethyl Diazoacetate to Give the Alkylating Agent and Mutagen Ethyl Chloro(hydroximino)acetate

Lin Zhou, James Haorah, Sheng C. Chen, Xiaojie Wang, Carol Kolar, Terence A. Lawson, Sidney S. Mirvish

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

Whereas nitrosation of secondary amines produces nitrosamines, amino acids with primary amino groups and glycine ethyl ester were reported to react with nitrite to give unidentified agents that alkylated 4-(p-nitrobenzyl)pyridine to produce purple dyes and be direct mutagens in the Ames test. We report here that treatment of glycine ethyl ester at 37 °C with excess nitrite acidified with HCl, followed by ether extraction, gave 30-40% yields of a product identified as ethyl chloro(hydroximino)acetate [ClC(=NOH)COOEt, ECHA] and a 9% yield of ethyl chloroacetate. The ECHA was identical to that synthesized by a known method from ethyl acetoacetate, strongly alkylated nitrobenzylpyridine, and may have arisen by N-nitrosation of glycine ethyl ester to give ethyl diazoacetate, which was C-nitrosated and reacted with chloride to give ECHA. Nitrosation of ethyl diazoacetate also yielded ECHA. Ethyl nitroacetate was not an intermediate as its nitrosation did not produce ECHA. ECHA reacted with aniline to give ethyl (hydroxamino)(phenylimino)acetate [PhN=C(NHOH)CO2Et]. This product was different from ethyl [(phenylamino)carbonyl]carbamate[PhNHC(=O)NHCO2Et], which was synthesized by reacting ethyl isocyanatoformate (OCN·CO2Et) with aniline. ECHA reacted with guanosine to give a derivative, which may have been a guanine-C(=NOH)CO2Et derivative. ECHA showed moderate toxicity and weak but significant mutagenicity without activation in Salmonella typhimurium TA-100 (mean, 1.31 × control value for 12-18 μg/plats) and for V79 mammalian cells (1.5-1.7 × control value for 60-100 μM). In conclusion, gastric nitrosation of glycine derivatives such as peptides with a N-terminal glycine might produce ECHA analogues that alkylate bases of gastric mucosal DNA and thereby initiate gastric cancer.

Original languageEnglish (US)
Pages (from-to)416-423
Number of pages8
JournalChemical Research in Toxicology
Volume17
Issue number3
DOIs
StatePublished - Mar 1 2004

ASJC Scopus subject areas

  • Toxicology

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