The Metabolism of Methazolamide in Immortalized Human Keratinocytes, HaCaT Cells.
The Metabolism of Methazolamide in Immortalized Human Keratinocytes, HaCaT Cells.
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DOI：
10.2174/1872312811666170127160931
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发表时间：
2017
影响因子：
--
通讯作者：
Sugiyama T
中科院分区：
文献类型：
--
作者：
Sasabe T;Maeda S;Kishida K;Yamano M;Miwa Y;Sugiyama T
药物治疗偶尔会伴有特异质的严重毒性，这种情况很少发生，但可能导致患者死亡。醋甲唑胺是一种抗青光眼药物，可引起严重的皮疹，称为Stevens-Johnson综合征/中毒性表皮坏死（SJS/TEN）。其确切病因仍不确定。在这项研究中，醋甲唑胺的代谢研究在永生化的人角质形成细胞，以揭示可能的机制，导致SJS/TEN。使用永生化人角质形成细胞HaCaT细胞研究醋甲唑胺的代谢。使用HPLC从培养基中分离代谢物。采用质谱法（LC-MS/MS）对其进行表征。研究了三种典型的化学诱导剂对细胞色素P450的诱导作用，并以甲巯咪唑作为黄素单加氧酶（FMO）的抑制剂。 最终代谢产物为磺酸N-[3-甲基-5-磺基-1，3，4-噻二唑-2（3 H）-亚基]乙酰胺（MSO）。地塞米松和β-萘啶酮在代谢中表现为细胞色素P450的诱导剂，而异烟肼则没有。甲巯咪唑的作用不一致。我们未检测到任何葡糖苷酸或巯基尿酸（N-乙酰半胱氨酸结合物）。 N-[3-甲基-5-磺基-1，3，4-噻二唑-2（3 H）-亚基]乙酰胺（MSO）不被认为是酶促反应的直接产物，而是N-[3-甲基-5-磺基-1，3，4-噻二唑-2（3 H）-亚基]乙酰胺的自动氧化产物，N-[3-甲基-5-磺基-1，3，4-噻二唑-2（3 H）-亚基]乙酰胺是一种化学不稳定的次磺酸，由细胞色素P450从醋甲唑酰胺的半胱氨酸缀合物的β-裂解酶产物产生。认为MSO对谷胱甘肽敏感，并返回醋甲唑胺的谷胱甘肽结合物，形成无效循环。一个假设的情况下提出的发病的疾病。
Drug therapy is occasionally accompanied by an idiosyncratic severe toxicity, which occurs very rarely, but can lead to patient mortality. Methazolamide, an anti-glaucomatous agent, could cause severe skin eruptions called Stevens-Johnson syndrome/toxic epidermal necrolyis (SJS/TEN). Its precise etiology is still uncertain. In this study, the metabolism of methazolamide was investigated in immortalized human keratinocytes to reveal the possible mechanism which causes SJS/TEN. The metabolism of methazolamide was studied using immortalized human keratinocytes, HaCaT cells. HPLC was used to isolate a metabolite from the culture medium. Mass spectrometry (LC-MS/MS) was employed for its characterization. Three typical chemical inducers were assessed for the inducibility of cytochrome P450, and methimazole was used as the inhibitor of flavin-containing monooxygenase (FMO). A sulfonic acid, N-[3-methyl-5-sulfo-1,3,4-thiadiazol-2(3H)-ylidene]acetamide (MSO) was identified as the final metabolite. Dexamethasone and β-naphthoflavone behaved as an inducer of cytochrome P450 in the metabolism, but isoniazid did not. The effect of methimazole was not consistent. We did not detect any glucuronide nor any mercapturic acid (N-acetylcysteine conjugate). N-[3-methyl-5-sulfo-1,3,4-thiadiazol-2(3H)-ylidene]acetamide (MSO) is not considered to be a direct product of an enzymatic reaction, but rather an auto-oxidation product of N-[3-methyl-5-sulfe-1,3,4-thiadiazol-2(3H)-ylidene]acetamide, a chemically unstable sulfenic acid, which is produced by cytochrome P450 from the β-lyase product of cysteine conjugate of methazolamide. MSO is considered to be susceptible to glutathione and to return to glutathione conjugate of methazolamide, forming a futile cycle. A hypothetical scenario is presented as to the onset of the disease.