Development and validation of a multiplex UHPLC‐MS/MS assay with stable isotopic internal standards for the monitoring of the plasma concentrations of the antiretroviral drugs bictegravir, cabotegravir, doravirine, and rilpivirine in people living with HIV

Abstract

The widespread use of highly active antiretroviral treatments has dramatically changed the prognosis of people living with HIV (PLWH). However, such treatments have to be taken lifelong raising issues regarding the maintenance of both therapeutic effectiveness and long-term tolerability. Recently approved or investigational antiretroviral drugs present considerable advantages, allowing once daily oral dosage along with activity against resistant variants (eg, bictegravir and doravirine) and also parenteral intramuscular administration that facilitates treatment adherence (eg, long-acting injectable formulations such as cabotegravir and rilpivirine). Still, there remains a risk of insufficient or exaggerated circulating exposure due to absorption issues, abnormal elimination, drugdrug interactions, and others. In this context, a multiplex ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) bioassay has been developed for the monitoring of plasma levels of bictegravir, cabotegravir, doravirine, and rilpivirine in PLWH. A simple and convenient protein precipitation was performed followed by direct injection of the supernatant into the UHPLC-MS/MS system.The four analytes were eluted in less than 3 minutes using a reversed-phase chromatography method coupled with triple quadrupole mass spectrometry detection. This bioassay was fully validated following international guidelines and achieved good performances in terms of trueness (94.7%-107.5%), repeatability (2.6%-11%), and intermediate precision (3.0%-11.2%) over the clinically relevant concentration ranges (from 30 to 9000 ng/mL for bictegravir, cabotegravir, and doravirine and from 10 to 1800 ng/mL for rilpivirine). This sensitive, accurate, and rapid UHPLC-MS/MS assay is currently applied in our laboratory for routine therapeutic drug monitoring of the oral drugs bictegravir and doravirine and is also intended to be applied for the monitoring of

Physiologically‐Based Pharmacokinetic Modeling Combined with Swiss HIV Cohort Study Data Supports No Dose Adjustment of Bictegravir in Elderly Individuals Living With HIV

“…Plasma was aliquoted and stored at −80°C until analysis by a validated liquid chromatography coupled to tandem mass spectrometry. 19 Measured bictegravir concentrations were combined within defined time intervals (0-2 hours, 2-4 hours, 4-6 hours, 6-8 hours, 8-12 hours, 12-16 hours, 16-20 hours, and 20-24 hours) to obtain a noncompartmental approximation of pharmacokinetic parameters in Matlab 2017a for young PLWH aged 20 to 55 years and elderly PLWH aged 55 to 85 years.…”

“…Blood samples were collected and centrifuged in EDTA‐containing tubes. Plasma was aliquoted and stored at −80°C until analysis by a validated liquid chromatography coupled to tandem mass spectrometry 19 …”

Tandem mass spectrometry of small-molecule antiviral drugs: 1. HIV-related antivirals

“… Compound m/z of [M+H] + Formula m/z of SRM product-ion Formula Literature dapivirine 330.171 [C 20 H 20 N 5 ] + 158.059 [C 8 H 6 N 4 ] +• ? [ 51 , 52 ] 119.086 [C 9 H 11 ] + [ 51 , 52 ] delavirdine 457.202 [C 22 H 29 N 6 O 3 S] + 221.176 [C 12 H 21 N 4 ] + [ 11 ] doravirine 426.058 [C 17 H 12 ClF 3 N 5 O 3 ] + 112.051 [C 4 H 6 N 3 O] + [ 53 ] 315.014 [C 13 H 7 ClF 3 N 2 O 2 ] + [ 53 ] efavirenz 316.035 [C 14 H 10 ClF 3 NO 2 ] + 244.014 [C 11 H 6 ClF 3 N] + [ 14 , 54 , 55 ] 232.014 …”

“…The major product ions of doravirine ([M+H] + with m/z 426) result from the cleavage of the C–N bond between the pyridin-2(1 H )-one moiety and the (4-methyl-5-oxo-4,5-dihydro-1 H -1,2,4-triazol-3-yl)methylene group and charge retention on either side, resulting in the complementary product ions with m/z 315 and 112, which have also been used in SRM [ 53 ]. The ion with m/z 315 shows subsequent losses of hydrogen fluoride (HF) to an ion with m/z 295 and a chlorine radical ( • Cl) to an ion with m/z 260.…”

In Silico Molecular Dynamics Docking of Drugs to the Inhibitory Active Site of SARS-CoV-2 Protease and Their Predicted Toxicology and ADME

“…R-428 (Rank 6, BE=-9.1, B12411) has been studied in experimental models of breast [59] , esophogeal [60] , and renal cell carcinoma [61] . Bictegravir (Rank 8, BE=-9.1, B11799) is an HIV-1 anti-retroviral developed by Gilead [62,63] . Ridinilazole (Rank 9, BE=-9, B15308) is a narrow-spectrum, non-absorbable antimicrobial with activity against Clostridium difficile undergoing clinical trials [64,65] .…”