Pediatric Pulmonology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
Cystic Fibrosis/Pulmonary Research Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
Pulm Pharmacol Ther. 2014 Feb;27(1):76-82. doi: 10.1016/j.pupt.2013.07.009. Epub 2013 Aug 7.
Although the airway surface is the anatomic target for many lung disease therapies, measuring drug concentrations and activities on these surfaces poses considerable challenges. We tested whether mass spectrometric analysis of exhaled breath condensate (EBC) could be utilized to non-invasively measure airway drug pharmacokinetics and predicted pharmacological activities. Mass spectrometric methods were developed to detect a novel epithelial sodium channel blocker (GS-9411/P-680), two metabolites, a chemically related internal standard, plus naturally occurring solutes including urea as a dilution marker. These methods were then applied to EBC and serum collected from four (Floridian) sheep before, during and after inhalation of nebulized GS-9411/P-680. Electrolyte content of EBC and serum was also assessed as a potential pharmacodynamic marker of drug activity. Airway surface concentrations of drug, metabolites, and electrolytes were calculated from EBC measures using EBC:serum urea based dilution factors. GS-9411/P-680 and its metabolites were quantifiable in the sheep EBC, with peak airway concentrations between 1.9 and 3.4 μM measured 1 h after inhalation. In serum, only Metabolite #1 was quantifiable, with peak concentrations ∼60-fold lower than those in the airway (45 nM at 1 h). EBC electrolyte concentrations suggested a pharmacological effect; but this effect was not statistical significant. Analysis of EBC collected during an inhalation drug study provided a method for quantification of airway drug and metabolites via mass spectrometry. Application of this methodology could provide an important tool in development and testing of drugs for airways diseases.
尽管气道表面是许多肺部疾病治疗的解剖学靶点,但测量这些表面的药物浓度和活性仍然存在很大的挑战。我们测试了呼气冷凝物(EBC)的质谱分析是否可以用于非侵入性地测量气道药物药代动力学和预测药理活性。开发了质谱方法来检测一种新型上皮钠通道阻滞剂(GS-9411/P-680)、两种代谢物、一种化学相关的内标以及包括尿素在内的天然存在的溶质,作为稀释标记物。然后将这些方法应用于 EBC 和血清,这些血清是在佛罗里达州的四只绵羊吸入雾化 GS-9411/P-680 之前、期间和之后采集的。EBC 和血清中的电解质含量也被评估为药物活性的潜在药效标志物。使用 EBC 中尿素的基于 EBC:血清的稀释因子,从 EBC 测量值中计算气道表面的药物、代谢物和电解质浓度。GS-9411/P-680 及其代谢物可在绵羊的 EBC 中定量,吸入后 1 小时测量到的气道峰浓度在 1.9 到 3.4 μM 之间。在血清中,只有代谢物 #1 可以定量,其峰值浓度比气道中的浓度低约 60 倍(1 小时时为 45 nM)。EBC 电解质浓度表明存在药理作用;但这种效果没有统计学意义。通过质谱分析对吸入药物研究中收集的 EBC 进行分析,提供了一种定量气道药物和代谢物的方法。该方法的应用可能为气道疾病药物的开发和测试提供重要工具。