Synthesis and Characterization of Methazolamide Conjugates

Methazolamide, acivicin, isoniazid and methimazole were purchased from Sigma Chemicals (St. Louis, MO, USA). Dexamethasone, β-naphthoflavone, dimethylsulfoxide (DMSO), acetic acid and acetonitrile were obtained from Wako Pure Chemicals (Osaka, Japan). In quantitative studies, we employed purified Methazolamide. It was purified in our laboratory by HPLC as described in the next paragraph. Cysteine, cysteinylglycine, and glutathione conjugates of Methazolamide were prepared in our laboratory according to the method described in the preceding paper [27 (link)]. N-[3 (link)-Methyl-5-sulfo-1,3,4-thiadiazol-2(3H)-ylidene]acetamide (MSO), and N-(3-methyl-5-mercapto-Δ⁴-1,3,4-thiadiazol–2-yl)acetamide (MSH) were also synthesized in our laboratory [14 (link)]. Fig. (1) shows the chemical structure of these compounds with their code names as used in this text. Phosphate-buffered saline (PBS) and Dulbecco’s MEM were obtained from Nikken Bioscience (Kyoto, Japan). Fetal bovine serum (Cat #12-10378) was supplied by IRH Biosciences (Lenexa, KS, USA).