Lalley-Chareczko Linden, Hiserodt Emily, Moorthy Ganesh, Zuppa Athena, Mounzer Karam, Koenig Helen
Philadelphia FIGHT Community Health Centers, Philadelphia, PA, United States.
Department of Anesthesiology and Critical Care Medicine, The Children's Hospital of Philadelphia, Philadelphia, PA, United States.
Front Pharmacol. 2020 Mar 19;11:286. doi: 10.3389/fphar.2020.00286. eCollection 2020.
HIV pre-exposure prophylaxis (PrEP) with tenofovir/emtricitabine is effective when taken daily. Previously, we developed a urine assay capable of detecting the prodrug tenofovir (TFV) in patients taking tenofovir disoproxil fumarate (TDF)-based PrEP. However, tenofovir alafenamide (TAF) has replaced TDF due to its different safety profile for HIV treatment and was recently approved as PrEP. Given the need to ensure the aforementioned assay remains available for the purpose of objective adherence monitoring, it is critical to ensure its accuracy for detecting TFV in patients taking TAF. Blood and urine samples were collected from 3 cohorts of patients: (1) 10 participants living with HIV (PLWH) with suppressed virus on a TAF-based regimen, (2) 10 HIV-participants administered 1 dose of TAF/FTC followed by urine and plasma sampling for 7 days starting 1-3 h post-dose, and (3) 10 HIV-participants administered 7 doses of TAF/FTC followed by urine and plasma sampling for 10 days starting 1-3 h after the last dose. Samples were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with high sensitivity and specificity for TFV. HIV-samples were compared to a historical cohort administered one dose of TDF/FTC. PLWH were 90% male, 40% African American, and 10% Hispanic (mean age = 57 y; SD 8.88 y). HIV-participants were 55% male and 70% Caucasian (mean age = 31.6 y; SD 7.70 y). Samples from PLWH demonstrated TFV concentrations 2 logs higher in urine than plasma (1,000 ng/mL vs ±10 ng/mL) at the time of collection. Urine samples following a single dose of TAF in HIV-participants yielded TFV concentrations ranging from 100 to 1,000 ng/mL 1-3 h post-dose and remained >100 ng/mL for 6 days in 8 of 10 participants. Urine samples collected after 7 consecutive doses of TAF yielded TFV concentrations >1,000 ng/mL 1-3 h after dosing discontinuation, with TFV concentrations >1,00 ng/mL 7 days post discontinuation in 8 of 10 participants. Urine TFV concentrations following TAF administration were comparable to those from a historical cohort administered TDF/FTC. Plasma TFV concentrations were low(±10 ng/mL) in both HIV-cohorts at all time points. TFV persists in urine at detectable concentrations in participants taking TAF/FTC for at least 7 days despite largely undetectable plasma concentrations, with urine TFV concentrations comparable to patients taking TDF/FTC. This study demonstrates the ability of a urine TFV assay to measure recent TAF adherence.
每日服用替诺福韦/恩曲他滨进行HIV暴露前预防（PrEP）是有效的。此前，我们开发了一种尿液检测方法，能够检测服用基于富马酸替诺福韦二吡呋酯（TDF）的PrEP的患者体内的前体药物替诺福韦（TFV）。然而，替诺福韦艾拉酚胺（TAF）因其在HIV治疗方面不同的安全性特征已取代TDF，并且最近被批准用于PrEP。鉴于需要确保上述检测方法可用于客观的依从性监测，确保其在检测服用TAF的患者体内TFV时的准确性至关重要。从3组患者中采集了血液和尿液样本：（1）10名接受基于TAF方案治疗且病毒得到抑制的HIV感染者（PLWH）；（2）10名未感染HIV的参与者，服用1剂TAF/ FTC，然后在给药后1 - 3小时开始进行7天的尿液和血浆采样；（3）10名未感染HIV的参与者，服用7剂TAF/ FTC，然后在最后一剂后1 - 3小时开始进行10天的尿液和血浆采样。使用对TFV具有高灵敏度和特异性的液相色谱 - 串联质谱法（LC - MS/MS）对样本进行分析。将未感染HIV的样本与接受1剂TDF/FTC的历史队列进行比较。PLWH中90%为男性，40%为非裔美国人，10%为西班牙裔（平均年龄 = 57岁；标准差8.88岁）。未感染HIV的参与者中55%为男性，70%为白种人（平均年龄 = 31.6岁；标准差7.70岁）。PLWH的样本在采集时尿液中TFV浓度比血浆中高2个对数（1000 ng/mL对±10 ng/mL）。未感染HIV的参与者服用单剂TAF后的尿液样本在给药后1 - 3小时产生的TFV浓度范围为100至1000 ng/mL，10名参与者中有8名在6天内TFV浓度保持>100 ng/mL。连续服用7剂TAF后采集的尿液样本在停药后1 - 3小时产生的TFV浓度>1000 ng/mL，10名参与者中有8名在停药7天后TFV浓度>100 ng/mL。服用TAF后尿液中的TFV浓度与接受TDF/FTC的历史队列中的浓度相当。在所有时间点，两个未感染HIV队列中的血浆TFV浓度都很低（±10 ng/mL）。尽管血浆浓度在很大程度上无法检测到，但服用TAF/FTC的参与者尿液中TFV以可检测浓度持续至少7天，尿液中TFV浓度与服用TDF/FTC的患者相当。这项研究证明了尿液TFV检测方法用于测量近期TAF依从性的能力。
