Department of Nutritional Sciences and Toxicology, 309 Morgan Hall, University of California, Berkeley, CA 94720-3104, USA.
J Pharmacol Exp Ther. 2010 Apr;333(1):281-9. doi: 10.1124/jpet.109.162487. Epub 2010 Jan 11.
Glucocorticoids are widely prescribed to treat autoimmune and inflammatory diseases. Although they are extremely potent, their utility in clinical practice is limited by a variety of adverse side effects. Development of compounds that retain the potent immunomodulating and anti-inflammatory properties of classic glucocorticoids while exhibiting reduced adverse actions is therefore a priority. Using heavy water labeling and mass spectrometry to measure fluxes through multiple glucocorticoid-responsive, disease-relevant target pathways in vivo in mice, we compared the effects of a classic glucocorticoid receptor (GR) ligand, prednisolone, with those of a novel arylpyrazole-based compound, L5 {[1-(4-fluorophenyl)-4a-methyl-5,6,7,8-tetrahydro-4H-benzo[f]indazol-5-yl]-[4-(trifluoromethyl)phenyl]methanol}. We show for the first time that L5 exhibits clearly selective actions on disease-relevant pathways compared with prednisolone. Prednisolone reduced bone collagen synthesis, skin collagen synthesis, muscle protein synthesis, and splenic lymphocyte counts, proliferation, and cell death, whereas L5 had none of those actions. In contrast, L5 was a more rapid and potent inhibitor of hippocampal neurogenesis than prednisolone, and L5 and prednisolone induced insulin resistance equally. Administration of prednisolone or L5 increased expression comparably for one GR-regulated gene involved in protein degradation in skeletal muscle (Murf1) and one GR-regulated gluconeogenic gene in liver (PEPCK). In summary, L5 dissociates the pleiotropic effects of the GR ligand prednisolone in intact animals in ways that neither gene expression nor cell-based models were able to fully capture or predict. Because multiple actions can be measured concurrently in a single animal, this method is a powerful systems approach for characterizing and differentiating the effects of ligands that bind nuclear receptors.
糖皮质激素被广泛用于治疗自身免疫性和炎症性疾病。尽管它们具有极强的功效，但由于各种不良反应，其在临床实践中的应用受到限制。因此，开发既能保留经典糖皮质激素强大的免疫调节和抗炎特性，又能减少不良反应的化合物，是当务之急。我们使用重水标记和质谱技术，在体内测量了小鼠中多个糖皮质激素反应性、与疾病相关的靶途径的通量，比较了经典糖皮质激素受体（GR）配体泼尼松龙和新型芳基吡唑基化合物 L5[1-(4-氟苯基)-4a-甲基-5,6,7,8-四氢-4H-苯并[f]吲哚-5-基]-[4-(三氟甲基)苯基]甲醇]的作用。我们首次表明，与泼尼松龙相比，L5 对与疾病相关的途径具有明显的选择性作用。泼尼松龙降低了骨胶原合成、皮肤胶原合成、肌肉蛋白合成和脾淋巴细胞计数、增殖和细胞死亡，而 L5 则没有这些作用。相反，L5 对海马神经发生的抑制作用比泼尼松龙更快、更强，而 L5 和泼尼松龙诱导胰岛素抵抗的作用则相等。泼尼松龙或 L5 的给药同样增加了骨骼肌中一个参与蛋白质降解的 GR 调节基因（Murf1）和肝脏中一个 GR 调节的糖异生基因（PEPCK）的表达。总之，L5 在完整动物中分离了 GR 配体泼尼松龙的多效作用，而基因表达或基于细胞的模型都无法完全捕捉或预测这些作用。由于可以在单个动物中同时测量多种作用，因此这种方法是一种强大的系统方法，可用于表征和区分结合核受体的配体的作用。
