在药物-药物相互作用(通常使用丙磺舒作为抑制剂)和药物-疾病相互作用的研究中,呋塞米(FUR)已用于体内评估肾转运蛋白OAT1和OAT3活性的探针药物混合物。本研究的目的是开发和验证血浆中FUR及其葡糖醛酸代谢物(FUR-GLU)分析的方法,血浆超滤液和尿液在药代动力学研究中的应用:孕妇的药物-药物相互作用试验研究(n=2),患者接受单次口服FUR(40mg),在另一种情况下,在单次口服FUR(40mg)之前接受单次口服丙磺舒(750mg),在非孕妇参与者中(n=12),仅接受单次口服FUR(40mg)。通过酸化的液-液萃取进行50μL血浆和血浆裂解物中FUR的样品制备,而50微升尿液和200微升血浆超滤液简单地用流动相稀释。该方法呈现的线性范围为0.50-2500ng/mL的血浆和血浆裂解物,0.125-250ng/mL血浆超滤液,和50-20,000ng/mL的尿液。FUR-GLU方法呈现的线性范围为0.125-250ng/mL的血浆超滤液和50-20,000ng/mL的尿液。精密度和准确度评价显示变异系数和相对误差<15%。在孕妇参与者中,FURCLRELER的平均值,CL分泌,地层。当丙磺舒与FUR一起给药时,FUR-GLU和CLnon肾脏均降低(8.24vs2.89L/h,8.15vs2.80L/h,3.86对1.75L/h,48.26对22.10L/h,分别)。非孕妇的FURCLRenal值相似,CL分泌,地层。FUR-GLU适用于仅接受FUR的孕妇。最后,未结合的FUR分数(fu)在孕妇中产生约1%的值,在非孕妇中产生0.22%的值。这些开发和验证的用于多种基质中的FUR和FUR-GLU定量的方法可以允许进一步研究当FUR作为OAT1和3体内探针施用时UGT1A9/1A1和fu。
Furosemide (FUR) has been used in probe drugs cocktails for in vivo evaluation of the renal transporters OAT1 and OAT3 activities in studies of drug-drug interactions (generally using probenecid as an inhibitor) and drug-disease interactions. The objective of this study was to develop and validate methods for FUR and its glucuronide metabolite (FUR-GLU) analysis in plasma, plasma ultrafiltrate and urine for application in pharmacokinetics studies: a pilot drug-drug interaction study in pregnant women (n = 2), who received a single oral dose of FUR (40 mg) and in another occasion a single oral dose of probenecid (750 mg) before a single oral dose of FUR (40 mg), and in non-pregnant women participants (n = 12), who only received a single oral dose of FUR (40 mg). The samples preparation for FUR in 50 µL of plasma and plasma lysate were carried by acidified liquid-liquid extraction, while 50 µL of urine and 200 µL of plasma ultrafiltrate were simply diluted with the mobile phase. The methods presented linearities in the range of 0.50 - 2500 ng/mL of plasma and plasma lysate, 0.125 - 250 ng/mL of plasma ultrafiltrate, and 50 - 20,000 ng/mL of urine. FUR-GLU methods presented linearities in the range of 0.125 - 250 ng/mL of plasma ultrafiltrate and 50 - 20,000 ng/mL of urine. Precision and accuracy evaluations showed coefficients of variation and relative errors < 15%. In the pregnant women participants, the mean values of FUR CLrenal, CLsecretion, CLformation. FUR-GLU and CLnon-renal were all reduced when probenecid was administered with FUR (8.24 vs 2.89 L/h, 8.15 vs 2.80 L/h, 3.86 vs 1.75 L/h, 48.26 vs 22.10 L/h, respectively). Non-pregnant women presented similar values of FUR CLrenal, CLsecretion, CLformation. FUR-GLU to the pregnant women who received FUR only. Finally, FUR fraction unbound (fu) resulted in values of approximately 1% in pregnant women and to 0.22% in non-pregnant women. These developed and validated methods for FUR and FUR-GLU quantification in multiple matrices can allow the further investigation of UGT1A9/1A1 and the fu when FUR is administered as an OAT 1 and 3 in vivo probe.