Linberg L, Wang J Z, Arison B H, Ulick S
Veterans Affairs Hospital, Bronx, NY 10468.
J Steroid Biochem Mol Biol. 1991 Mar;38(3):351-7. doi: 10.1016/0960-0760(91)90107-g.
11 beta-Hydroxy dehydrogenation of cortisol to cortisone is specifically impaired in the syndrome of apparent mineralocorticoid excess. This defect bears on the pathogenesis of the disorder by unmasking the potential mineralocorticoid agonism of unmetabolized cortisol at or near mineralocorticoid target tissues. A specific index of this defect is provided by measurement of the formation of tritiated water following the administration of [3H]11 alpha-cortisol. We have explored the use of a non-radioactive tracer to follow this unidirectional dehydrogenation reaction but because of the relatively lower sensitivity of measurement of 2H2O compared to 3H2O in body fluids, use of the corresponding [2H]11 alpha-cortisol was not feasible. We have devised instead a method incorporating additional deuterium atoms into cortisol to measure unidirectional 11 beta-hydroxy dehydrogenation not by the formation of labeled water but by the determination of the dehydrogenated cortisol product from its residual deuterium content. Cortisol-d4 metabolized to cortisone-d3 is conveniently measured by the techniques of organic mass spectrometry. The synthesis of cortisol-9 alpha, 11 alpha, 12 alpha 12 beta-d4 and the validation of its isotopic distribution by mass spectrometry and nuclear magnetic resonance is described.
在表观盐皮质激素过多综合征中，皮质醇向可的松的11β-羟基脱氢反应存在特异性受损。这种缺陷通过在盐皮质激素靶组织或其附近暴露未代谢皮质醇潜在的盐皮质激素激动作用，对该疾病的发病机制产生影响。通过测量给予[3H]11α-皮质醇后氚化水的形成，可提供这种缺陷的一个特异性指标。我们探索了使用非放射性示踪剂来追踪这种单向脱氢反应，但由于与体液中3H2O相比，2H2O测量的灵敏度相对较低，因此使用相应的[2H]11α-皮质醇并不可行。相反，我们设计了一种方法，将额外的氘原子引入皮质醇中，以通过从其残留氘含量测定脱氢皮质醇产物来测量单向11β-羟基脱氢，而不是通过标记水的形成来测量。通过有机质谱技术可方便地测量代谢为可的松-d3的皮质醇-d4。描述了皮质醇-9α,11α,12α,12β-d4的合成及其通过质谱和核磁共振对其同位素分布的验证。
