Studies of chemical selectivity of hapten, reactivity, and skin sensitization potency. 3. Synthesis and studies on the reactivity toward model nucleophiles of the 13C-labeled skin sensitizers, 5-chloro-2-methylisothiazol-3-one (MCI) and 2-methylisothiazol-3-one (MI).
The skin sensitizers, 5-chloro-2-methylisothiazol-3-one (MCI) and 2-methylisothiazol-3-one (MI), have been synthesized isotopically labeled with (13)C at all carbon positions. The reactivity of 3-[(13)C]-, 4-[(13)C]-, and 5-[(13)C]MCI and MI toward a series of model nucleophiles for protein amino acid residues, i.e., butylamine, imidazole, sodium propanethiolate, and sodium phenoxide, was followed by (13)C and (1)H[(13)C] NMR spectroscopy. While MCI was found to react quantitatively with sodium propanethiolate and butylamine and significantly with imidazole and sodium phenoxide, MI reacted only with sodium propanethiolate. Reaction of MCI with nonthiol nucleophiles proceeded through an initial addition-elimination at position 5, leading to stable substitution adducts in the case of imidazole and sodium phenoxide. In the case of butylamine, the initial adduct was subjected to extra reactions at the sulfur atom through a cleavage of the S-N bond, leading to open adducts of the thioamide or amide type. Experiments carried out with N-acetyl-Cys, in excess or in deficiency, indicated that thiol nucleophiles reacted first at the sulfur atom through a cleavage of the S-N bond followed by extra nucleophilic reactions leading to open adducts of the mercaptothioester or mercaptoester type. Reaction of MCI with thiol nucleophiles gave products consistent with the formation of a reactive thioacyl chloride intermediate able to react with other nucleophiles present in the reaction medium. As a consequence, N-acetyl-Cys was found to be able to activate MCI toward N(alpha)-acetyl-Lys under physiological conditions to form adducts of the thioamide or amide type. Thus MCI, a strong sensitizer, and MI, a weak sensitizer, were found to react with different nucleophiles through different mechanisms. Although both MCI and MI can react with thiol nucleophiles, only MCI is capable of significantly reacting with amino nucleophiles of the Lys or His type. Moreover, MCI could be activated by a prior reaction with thiols.
Alvarez-Sánchez
Rubén
R
Laboratoire de Dermatochimie (UMR 7123), Université Louis Pasteur, Clinique Dermatologique, CHU, Strasbourg, France.
Basketter
David
D
Pease
Camilla
C
Lepoittevin
Jean-Pierre
JP
D002247:Carbon Isotopes; D006241:Haptens / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D012867:Skin; D013844:Thiazoles / Q000138:chemical synthesis*
D002247:Carbon Isotopes; D006241:Haptens; D013844:Thiazoles; C011506:2-methyl-4-isothiazolin-3-one; C011421:5-chloro-2-methyl-4-isothiazolin-3-one