26 results on '"Schauer, Amanda P."'
Search Results
2. Ultra-long-acting in-situ forming implants with cabotegravir protect female macaques against rectal SHIV infection
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Young, Isabella C., Massud, Ivana, Cottrell, Mackenzie L., Shrivastava, Roopali, Maturavongsadit, Panita, Prasher, Alka, Wong-Sam, Andres, Dinh, Chuong, Edwards, Tiancheng, Mrotz, Victoria, Mitchell, James, Seixas, Josilene Nascimento, Pallerla, Aryani, Thorson, Allison, Schauer, Amanda, Sykes, Craig, De la Cruz, Gabriela, Montgomery, Stephanie A., Kashuba, Angela D. M., Heneine, Walid, Dobard, Charles W., Kovarova, Martina, Garcia, J. Victor, Garcίa-Lerma, J. Gerardo, and Benhabbour, S. Rahima
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- 2023
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3. HIV and urethritis: time required for antiretroviral therapy to suppress HIV in semen
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Matoga, Mitch, Chen, Jane S., Massa, Cecilia, Thengolose, Isaac, Tegha, Gerald, Ndalama, Beatrice, Bonongwe, Naomi, Mathiya, Esther, Jere, Edward, Banda, Gabriel, Khan, Shiraz, Loftis, Amy J., Kashuba, Angela, Cottrell, Mackenzie L., Schauer, Amanda P., Van Horne, Brian, Tompkins, Lauren A., Lancaster, Kathryn E., Miller, William C., Eron, Joseph J., Hoffman, Irving F., and Cohen, Myron S.
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- 2023
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4. Validation of an LC-MS/MS assay for the simultaneous determination of bictegravir, doravirine, and raltegravir in human plasma
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Schauer, Amanda P., Sykes, Craig, Cottrell, Mackenzie L., Imaz, Arkaitz, Podzamczer, Daniel, and Kashuba, Angela DM
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- 2022
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5. A long-acting formulation of rifabutin is effective for prevention and treatment of Mycobacterium tuberculosis
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Kim, Manse, Johnson, Claire E., Schmalstig, Alan A., Annis, Ayano, Wessel, Sarah E., Van Horn, Brian, Schauer, Amanda, Exner, Agata A., Stout, Jason E., Wahl, Angela, Braunstein, Miriam, Victor Garcia, J., and Kovarova, Martina
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- 2022
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6. Next-Generation Contraceptive Intravaginal Ring: Comparison of Etonogestrel and Ethinyl Estradiol In Vitro and In Vivo Release from 3D-Printed Intravaginal Ring and NuvaRing.
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Young, Isabella C., Thorson, Allison L., Cottrell, Mackenzie L., Sykes, Craig, Schauer, Amanda P., Sellers, Rani S., Janusziewicz, Rima, Vincent, Kathleen L., and Benhabbour, Soumya Rahima
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VAGINAL contraceptives ,ETHINYL estradiol ,DRUG delivery systems ,THREE-dimensional printing ,LONG-acting reversible contraceptives ,VAGINAL medication - Abstract
Intravaginal rings (IVRs) represent a well-established, woman-controlled and sustained vaginal drug delivery system suitable for a wide range of applications. Here, we sought to investigate the differences in etonogestrel (ENG) and ethinyl estradiol (EE) release from a 3D-printed IVR utilizing continuous liquid interface production (CLIP™) (referred to as CLIP
LOW for low drug loading and CLIPHIGH IVRs for high drug loading) and NuvaRing, a commercially available injection molded IVR. We conducted in vitro release studies in simulated vaginal fluid to compare the release of ENG and EE from CLIPLOW IVRs and NuvaRing. CLIPLOW IVRs had a similar hormone dose to NuvaRing and exhibited slightly slower ENG release and greater EE release in vitro compared to NuvaRing. When administered to female sheep, NuvaRing demonstrated greater ENG/EE levels in plasma, vaginal tissue and vaginal fluids compared to CLIPLOW IVR despite similar drug loadings. Leveraging observed hormones levels in sheep from NuvaRing as an effective contraceptive benchmark, we developed a long-acting CLIPHIGH IVR with increased ENG and EE doses that demonstrated systemic and local hormone levels greater than the NuvaRing for 90 days in sheep. No signs of toxicity were noted regarding general health, colposcopy, or histological analysis in sheep after CLIPHIGH IVR administration. Our results provided (1) a comparison of ENG and EE release between a 3D-printed IVR and NuvaRing in vitro and in vivo, (2) a preclinical pharmacokinetic benchmark for vaginally delivered ENG and EE and (3) the generation of a 90-day CLIP IVR that will be utilized in future work to support the development of a long-acting ENG/EE IVR combined with an antiretroviral for the prevention of HIV and unplanned pregnancy. [ABSTRACT FROM AUTHOR]- Published
- 2024
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7. Pregnancy-Related Hormones Increase Nifedipine Metabolism in Human Hepatocytes by Inducing CYP3A4 Expression
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Khatri, Raju, Kulick, Natasha, Rementer, Rebecca J.B., Fallon, John K., Sykes, Craig, Schauer, Amanda P., Malinen, Melina M., Mosedale, Merrie, Watkins, Paul B., Kashuba, Angela D.M., Boggess, Kim A., Smith, Philip C., Brouwer, Kim L.R., and Lee, Craig R.
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- 2021
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8. Validation of an LC–MS/MS assay to simultaneously monitor the intracellular active metabolites of tenofovir, emtricitabine, and lamivudine in dried blood spots
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Schauer, Amanda P., Sykes, Craig, Cottrell, Mackenzie L., Prince, Heather, and Kashuba, Angela D.M.
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- 2018
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9. Development and validation of an LC-MS/MS assay for the quantification of dolutegravir extracted from human hair
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Sykes, Craig, Blake, Kimberly, White, Nicole, Schauer, Amanda P, Guzman, Bryan B., Cottrell, Mackenzie L., Tamraz, Bani, and Kashuba, Angela D. M.
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- 2018
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10. Pharmacokinetic Study of Islatravir and Etonogestrel Implants in Macaques.
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Daly, Michele B., Wong-Sam, Andres, Li, Linying, Krovi, Archana, Gatto, Gregory J., Norton, Chasity, Luecke, Ellen H., Mrotz, Victoria, Forero, Catalina, Cottrell, Mackenzie L., Schauer, Amanda P., Gary, Joy, Nascimento-Seixas, Josilene, Mitchell, James, van der Straten, Ariane, Heneine, Walid, Garcίa-Lerma, J. Gerardo, Dobard, Charles W., and Johnson, Leah M.
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LONG-acting reversible contraceptives ,BIOABSORBABLE implants ,PHARMACOKINETICS ,MACAQUES ,UNPLANNED pregnancy ,HIV prevention ,PREGNANCY tests ,CONTROLLED release drugs - Abstract
The prevention of HIV and unintended pregnancies is a public health priority. Multi-purpose prevention technologies capable of long-acting HIV and pregnancy prevention are desirable for women. Here, we utilized a preclinical macaque model to evaluate the pharmacokinetics of biodegradable ε-polycaprolactone implants delivering the antiretroviral islatravir (ISL) and the contraceptive etonogestrel (ENG). Three implants were tested: ISL-62 mg, ISL-98 mg, and ENG-33 mg. Animals received one or two ISL-eluting implants, with doses of 42, 66, or 108 µg of ISL/day with or without an additional ENG-33 mg implant (31 µg/day). Drug release increased linearly with dose with median [range] plasma ISL levels of 1.3 [1.0–2.5], 1.9 [1.2–6.3] and 2.8 [2.3–11.6], respectively. The ISL-62 and 98 mg implants demonstrated stable drug release over three months with ISL-triphosphate (ISL-TP) concentr54ations in PBMCs above levels predicted to be efficacious for PrEP. Similarly, ENG implants demonstrated sustained drug release with median [range] plasma ENG levels of 495 [229–1110] pg/mL, which suppressed progesterone within two weeks and showed no evidence of altering ISL pharmacokinetics. Two of the six ISL-98 mg implants broke during the study and induced implant-site reactions, whereas no reactions were observed with intact implants. We show that ISL and ENG biodegradable implants are safe and yield sufficient drug levels to achieve prevention targets. The evaluation of optimized implants with increased mechanical robustness is underway for improved durability and vaginal efficacy in a SHIV challenge model. [ABSTRACT FROM AUTHOR]
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- 2023
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11. Pregnancy related hormones increase CYP3A mediated buprenorphine metabolism in human hepatocytes: a comparison to CYP3A substrates nifedipine and midazolam.
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Fashe, Muluneh M., Miner, Taryn A., Fallon, John K., Schauer, Amanda P., Sykes, Craig, Smith, Philip C., and Lee, Craig R.
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CYTOCHROME P-450 CYP3A ,NIFEDIPINE ,MIDAZOLAM ,LIVER cells ,HORMONES ,BUPRENORPHINE ,METABOLISM - Abstract
Introduction: Pregnancy increases the clearance of CYP3A4 substrate drugs and pregnancy-related hormones (PRHs) induce hepatic CYP3A4 expression and metabolism. However, it remains unclear to what extent the magnitude of PRHevoked changes in hepatic CYP3A metabolism varies across multiple substrates. This study quantified the impact of PRHs on CYP3A protein concentrations and buprenorphine metabolism in human hepatocytes, and compared the magnitude of these effects to nifedipine and midazolam metabolism. Methods: Sandwich-cultured human hepatocytes (SCHH) from female donors were exposed to PRHs, administered in combination across a range of physiologically relevant concentrations, for 72 h. Absolute protein concentrations of CYP3A4, CYP3A5, and CYP3A7 in SCHH membrane fractions were quantified by nanoLC-MS/MS, and norbuprenorphine (nor-BUP), dehydronifedipine (dehydro-NIF), and 1-hydroxy-midazolam (1-OH-MDZ) formation was evaluated. Results: Compared to control, PRH exposure increased CYP3A4, CYP3A7, and total CYP3A protein concentrations, but not CYP3A5 concentrations, and increased nor-BUP, dehydro-NIF, and 1-OH-MDZ formation in a concentration-dependent manner. The formation of nor-BUP, dehydro-NIF, and 1-OH-MDZ each positively correlated with PRH-mediated changes in total CYP3A protein concentrations. The PRH-evoked increase in nor-BUP formation was evident in all donors; however, the PRH induction of dehydro-NIF and 1-OH-MDZformationwas diminished in a hepatocyte donorwith high basal CYP3A5 expression. Discussion: These findings demonstrate that PRHs increase buprenorphine, nifedipine, and midazolam metabolism in SCHH via induction of CYP3A4 and total CYP3A protein concentrations, and the magnitude of these effects vary across hepatocyte donors in a substrate-specific manner. These data provide insight into the contribution of PRH induction of CYP3A4 metabolism to increased buprenorphine clearance during pregnancy. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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12. Dose-Ranging Plasma and Genital Tissue Pharmacokinetics and Biodegradation of Ultra-Long-Acting Cabotegravir In Situ Forming Implant.
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Young, Isabella C., Thorson, Allison L., Shrivastava, Roopali, Sykes, Craig, Schauer, Amanda P., Cottrell, Mackenzie L., Kashuba, Angela D. M., and Benhabbour, Soumya Rahima
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POLYMER degradation ,PHARMACOKINETICS ,HIV prevention ,BIODEGRADATION ,MICE ,TISSUES - Abstract
HIV continues to affect millions of men and women worldwide. The development of long-acting injectables for HIV prevention can overcome adherence challenges with daily oral prevention regimens by reducing dosing frequency and stigma. We previously developed an ultra-long-acting injectable, biodegradable, and removeable in situ forming implant (ISFI) with cabotegravir (CAB) that demonstrated protection after multiple rectal SHIV challenges in female macaques. Here, we sought to further characterize CAB ISFI pharmacokinetics (PK) in mice by assessing the effect of dose and number of injections on CAB PK, time to completion of CAB release and polymer degradation, long-term genital tissue PK, and CAB PK tail after implant removal. CAB concentrations in plasma were above the benchmark for protection for 11–12 months with proportionality between dose and drug exposure. CAB ISFI exhibited high concentrations in vaginal, cervical, and rectal tissues for up to 180 days. Furthermore, depots were easily retrievable up to 180 days post-administration with up to 34% residual CAB and near complete (85%) polymer degradation quantified in depots ex vivo. After depot removal, results demonstrated a median 11-fold decline in CAB plasma concentrations across all doses. Ultimately, this study provided critical PK information for the CAB ISFI formulation that could aid in its future translation to clinical studies. [ABSTRACT FROM AUTHOR]
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- 2023
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13. A Novel Algorithm to Improve PrEP Adherence Monitoring Using Dried Blood Spots.
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Devanathan, Aaron S., Dumond, Julie B., Anderson, Daijha J.C., Moody, Kristen, Poliseno, Amanda J., Schauer, Amanda P., Sykes, Craig, Gay, Cynthia L., Rosen, Elias P., Kashuba, Angela D.M., and Cottrell, Mackenzie L.
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DIRECTLY observed therapy ,PRE-exposure prophylaxis ,ALGORITHMS ,DRIED blood spot testing ,PROOF of concept - Abstract
Tenofovir diphosphate (TFVdp; an active metabolite of oral HIV pre‐exposure prophylaxis (PrEP)) is measured in dried blood spots (DBS) to estimate adherence. However, TFVdp's long half‐life in whole blood may lead to misclassification following a recent change in adherence. PrEP's other metabolite, emtricitabine triphosphate (FTCtp), has a shorter half‐life in whole blood but adherence thresholds are undefined. We characterized DBS TFVdp and FTCtp concentrations across many dosing scenarios. Population pharmacokinetic models were fit to TFVdp and FTCtp DBS concentrations from a directly observed therapy study (NCT03218592). Concentrations were simulated for 90 days of daily dosing followed by 90 days of 1 to 7 doses/week and for event‐driven PrEP (edPrEP) scenarios. Thresholds of 1,000 and 200 fmol/punch, for TFVdp and FTCtp, respectively, were reflective of taking 4 doses/week (a minimum target for effective PrEP in men). TFVdp was < 1,000 fmol/punch for 17 days after initiating daily PrEP and > 1,000 fmol/punch for 62 days after decreasing to 3 doses/week. Respectively, FTCtp was < 200 fmol/punch for 4 days and > 200 fmol/punch for 6 days. Accuracy of edPrEP adherence classification depended on duration between last sex act and DBS sampling for both measures with misclassification ranging from 9–100%. These data demonstrate adherence misclassification by DBS TFVdp for 2 months following a decline in adherence, elucidating the need for FTCtp to estimate recent adherence. We provide proof of principle that individualized interpretation is needed to support edPrEP adherence monitoring. Our collective approach facilitates clinicians' ability to interpret DBS results and administer patient‐centric interventions. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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14. A quantitative LC–MS/MS method for the determination of tissue brincidofovir and cidofovir diphosphate in a MuPyV‐infected mouse model.
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Guzman, Bryan B., Schauer, Amanda P., Dunn, John A., Cottrell, Mackenzie L., and Sykes, Craig
- Abstract
Brincidofovir (BCV) is an investigational lipid conjugate of the nucleotide analog cidofovir (CDV), which is being developed as a medical countermeasure for the treatment of smallpox. BCV is active against double‐stranded DNA viruses including BK and JC viruses. Here, we validated procedures for quantifying BCV and its pharmacologically active moiety cidofovir diphosphate (CDV‐PP) in mouse kidney, brain and spleen tissue homogenates. Following homogenization, BCV and CDV‐PP were extracted from the tissues by protein precipitation with their stable, isotopically labeled internal standards, BCV‐d6 and 13C315N2‐CDV‐PP. Then, samples were analyzed for BCV by reverse‐phase chromatography on a Waters Xterra MS C18 (50 × 2.1 mm, 3.5 μm particle size) column while CDV‐PP was analyzed on a Thermo BioBasic AX (50 × 2.1 mm, 5 μm particle size) column using anion exchange chromatography. Detection was achieved by electrospray ionization in positive ion mode on an AB Sciex API‐5000 triple quadrupole mass spectrometer. The calibration curves were linear over a range of 1.00–1,000 ng/ml homogenate and 0.050–50.0 ng/ml homogenate for BCV and CDV‐PP, respectively. These methods were validated according to US Food and Drug Administration guidance for industry and may be used to characterize the tissue pharmacology of both analytes to advance its preclinical development. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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15. Decreased Tenofovir Diphosphate Concentrations in a Transgender Female Cohort: Implications for Human Immunodeficiency Virus Preexposure Prophylaxis.
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Cottrell, Mackenzie L, Prince, Heather M A, Schauer, Amanda P, Sykes, Craig, Maffuid, Kaitlyn, Poliseno, Amanda, Chun, Tae-Wook, Huiting, Erin, Stanczyk, Frank Z, Peery, Anne F, Dellon, Evan S, Adams, Jessica L, Gay, Cindy, and Kashuba, Angela D M
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HIV prevention ,TISSUE analysis ,BLOOD testing ,CLINICAL trials ,COMPARATIVE studies ,DRUG interactions ,ESTRADIOL ,HIV-positive persons ,SEX hormones ,PREVENTIVE medicine ,WOMEN ,ANDROGEN-insensitivity syndrome ,TREATMENT effectiveness ,TENOFOVIR ,PHARMACODYNAMICS - Abstract
Feminizing hormone therapy (FHT) may interact with human immunodeficiency virus preexposure prophylaxis (PrEP). We found that transgender women who took FHT exhibited a 7-fold lower rectal tissue ratio of PrEP's active metabolites vs competing deoxynucleotides compared to cisgender women and men (P =.03) that inversely correlated with estradiol (ρ = –0.79; P <.05). Thus, FHT may negatively impact PrEP efficacy. Clinical Trials Registration. NCT02983110. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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16. Population Modeling Highlights Drug Disposition Differences Between Tenofovir Alafenamide and Tenofovir Disoproxil Fumarate in the Blood and Semen.
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Greene, Stephen A., Chen, Jingxian, Prince, Heather M. A., Sykes, Craig, Schauer, Amanda P., Blake, Kimberly, Nelson, Julie A. E., Gay, Cynthia L., Cohen, Myron S., and Dumond, Julie B.
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MALE reproductive organs ,SEMEN ,TENOFOVIR ,BISOPROLOL ,BLOOD plasma ,DRUG metabolism ,GLYCOPYRROLATE - Abstract
Understanding antiretroviral disposition in the male genital tract, a distinct viral compartment, can provide insight for the eradication of HIV. Population pharmacokinetic modeling was conducted to investigate the disposition of tenofovir disoproxil fumarate (TDF), tenofovir alafenamide (TAF), and emtricitabine and their metabolites in blood and semen. Blood plasma and seminal plasma (SP) concentrations of tenofovir and emtricitabine were measured, as were tenofovir‐diphosphate and emtricitabine‐triphosphate concentrations in peripheral blood mononuclear cells (PBMCs) and seminal mononuclear cells. Sequential compartmental modeling described drug disposition in blood and semen. Our modeling suggests slower elimination of apparent tenofovir‐diphosphate PBMC and faster elimination of tenofovir SP after administration of TAF compared with TDF, likely reflecting flip‐flop kinetics. Additionally, TAF metabolism to tenofovir appeared slower in semen compared with blood; however, SP elimination of TAF‐derived tenofovir appeared faster than its blood plasma elimination. These findings provide valuable insight for further mechanistic study of cellular entry and drug metabolism in the male genital tract. [ABSTRACT FROM AUTHOR]
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- 2019
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17. Antiretroviral concentrations and surrogate measures of efficacy in the brain tissue and CSF of preclinical species.
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Srinivas, Nithya, Rosen, Elias P., Gilliland Jr, William M., Kovarova, Martina, Remling-Mulder, Leila, De La Cruz, Gabriela, White, Nicole, Adamson, Lourdes, Schauer, Amanda P., Sykes, Craig, Luciw, Paul, Garcia, J. Victor, Akkina, Ramesh, and Kashuba, Angela D. M.
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CEREBROSPINAL fluid ,BRAIN ,RHESUS monkeys ,PROTEIN binding ,TISSUES ,AUTOPSY - Abstract
1. Antiretroviral concentrations in cerebrospinal fluid (CSF) are used as surrogate for brain tissue, although sparse data support this. We quantified antiretrovirals in brain tissue across preclinical models, compared them to CSF, and calculated 90% inhibitory quotients (IQ
90 ) for nonhuman primate (NHP) brain tissue. Spatial distribution of efavirenz was performed by mass-spectrometry imaging (MSI). 2. HIV or RT-SHIV-infected and uninfected animals from two humanized mouse models (hemopoietic-stem cell/RAG2-, n = 36; bone marrow-liver-thymus/BLT, n =13) and an NHP model (rhesus macaque, n =18) were dosed with six antiretrovirals. Brain tissue, CSF (NHPs), and plasma were collected at necropsy. Drug concentrations were measured by LC-MS/MS. Rapid equilibrium dialysis determined protein binding in NHP brain. 3. Brain tissue penetration of most antiretrovirals were >10-fold lower (p < 0.02) in humanized mice than NHPs. NHP CSF concentrations were >13-fold lower (p <0.02) than brain tissue with poor agreement except for efavirenz (r = 0.91, p = 0.001). Despite 97% brain tissue protein binding, efavirenz achieved IQ90 >1 in all animals and 2-fold greater white versus gray matter concentration. 4. Brain tissue penetration varied across animal models for all antiretrovirals except raltegravir, and extrapolating brain tissue concentrations between models should be avoided. With the exception of efavirenz, CSF is not a surrogate for brain tissue concentrations. [ABSTRACT FROM AUTHOR]- Published
- 2019
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18. Predicting Efavirenz Concentrations in the Brain Tissue of HIV‐Infected Individuals and Exploring their Relationship to Neurocognitive Impairment.
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Srinivas, Nithya, Joseph, Sarah Beth, Robertson, Kevin, Kincer, Laura P., Menezes, Prema, Adamson, Lourdes, Schauer, Amanda P., Blake, Kimberly H., White, Nicole, Sykes, Craig, Luciw, Paul, Eron, Joseph J., Forrest, Alan, Price, Richard W., Spudich, Serena, Swanstrom, Ronald, and Kashuba, Angela D.M.
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CEREBROSPINAL fluid ,EFAVIRENZ ,BRAIN diseases ,CENTRAL nervous system ,PHARMACOKINETICS - Abstract
Sparse data exist on the penetration of antiretrovirals into brain tissue. In this work, we present a framework to use efavirenz (EFV) pharmacokinetic (PK) data in plasma, cerebrospinal fluid (CSF), and brain tissue of eight rhesus macaques to predict brain tissue concentrations in HIV‐infected individuals. We then perform exposure‐response analysis with the model‐predicted EFV area under the concentration‐time curve (AUC) and neurocognitive scores collected from a group of 24 HIV‐infected participants. Adult rhesus macaques were dosed daily with 200 mg EFV (as part of a four‐drug regimen) for 10 days. Plasma was collected at 8 time points over 10 days and at necropsy, whereas CSF and brain tissue were collected at necropsy. In the clinical study, data were obtained from one paired plasma and CSF sample of participants prescribed EFV, and neuropsychological test evaluations were administered across 15 domains. PK modeling was performed using ADAPT version 5.0 Biomedical Simulation Resource, Los Angeles, CA) with the iterative two‐stage estimation method. An eight‐compartment model best described EFV distribution across the plasma, CSF, and brain tissue of rhesus macaques and humans. Model‐predicted median brain tissue concentrations in humans were 31 and 8,000 ng/mL, respectively. Model‐predicted brain tissue AUC was highly correlated with plasma AUC (γ = 0.99, P < 0.001) but not CSF AUC (γ = 0.34, P = 0.1) and did not show any relationship with neurocognitive scores (γ < 0.05, P > 0.05). This analysis provides an approach to estimate PK the brain tissue in order to perform PK/pharmacodynamic analyses at the target site. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
19. Pregnancy-Related Hormones Increase UGT1A1-Mediated Labetalol Metabolism in Human Hepatocytes.
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Khatri, Raju, Fallon, John K., Sykes, Craig, Kulick, Natasha, Rementer, Rebecca J. B., Miner, Taryn A., Schauer, Amanda P., Kashuba, Angela D. M., Boggess, Kim A., Brouwer, Kim L. R., Smith, Philip C., and Lee, Craig R.
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LIVER cells ,URIDINE diphosphate ,METABOLISM ,CYTOCHROME P-450 ,HORMONES - Abstract
Pregnancy-related hormones (PRH) are recognized as important regulators of hepatic cytochrome P450 enzyme expression and function. However, the impact of PRH on the hepatic expression and function of uridine diphosphate glucuronosyltransferases (UGTs) remains unclear. Using primary human hepatocytes, we evaluated the effect of PRH exposure on mRNA levels and protein concentrations of UGT1A1, UGT2B7, and other key UGT enzymes, and on the metabolism of labetalol (a UGT1A1 and UGT2B7 substrate commonly prescribed to treat hypertensive disorders of pregnancy). Sandwich-cultured human hepatocytes (SCHH) from female donors were exposed to the PRH estradiol, estriol, estetrol, progesterone, and cortisol individually or in combination. We quantified protein concentrations of UGT1A1, UGT2B7, and four additional UGT1A isoforms in SCHH membrane fractions and evaluated the metabolism of labetalol to its glucuronide metabolites in SCHH. PRH exposure increased mRNA levels and protein concentrations of UGT1A1 and UGT1A4 in SCHH. PRH exposure also significantly increased labetalol metabolism to its UGT1A1-derived glucuronide metabolite in a concentration-dependent manner, which positively correlated with PRH-induced changes in UGT1A1 protein concentrations. In contrast, PRH did not alter UGT2B7 mRNA levels or protein concentrations in SCHH, and formation of the UGT2B7-derived labetalol glucuronide metabolite was decreased following PRH exposure. Our findings demonstrate that PRH alter expression and function of UGT proteins in an isoform-specific manner and increase UGT1A1-mediated labetalol metabolism in human hepatocytes by inducing UGT1A1 protein concentrations. These results provide mechanistic insight into the increases in labetalol clearance observed in pregnant individuals. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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20. Altered Response Pattern following AZD5582 Treatment of SIVInfected, ART-Suppressed Rhesus Macaque Infants.
- Author
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Bricker, Katherine M., Obregon-Perko, Veronica, Williams, Brianna, Oliver, Danielle, Uddin, Ferzan, Neja, Margaret, Hopkins, Louis, Dashti, Amir, Jean, Sherrie, Wood, Jennifer S., Ehnert, Stephanie, Shan Liang, Vanderford, Thomas, Tharp, Gregory K., Bosinger, Steven E., Schauer, Amanda P., Mavigner, Maud, Cottrell, Mackenzie L., Margolis, David, and Dunham, Richard M.
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RHESUS monkeys , *MACAQUES , *INFANTS , *VIRUS reactivation , *T cells , *HIV-positive children , *ANTIRETROVIRAL agents - Abstract
The “shock and kill” strategy for HIV-1 cure incorporates latency-reversing agents (LRA) in combination with interventions that aid the host immune system in clearing virally reactivated cells. LRAs have not yet been investigated in pediatric clinical or preclinical studies. Here, we evaluated an inhibitor of apoptosis protein (IAP) inhibitor (IAPi), AZD5582, that activates the noncanonical NF-k B (ncNF-k B) signaling pathway to reverse latency. Ten weekly doses of AZD5582 were intravenously administered at 0.1 mg/kg to rhesus macaque (RM) infants orally infected with SIVmac251 at 4 weeks of age and treated with a triple ART regimen for over 1 year. During AZD5582 treatment, on-ART viremia above the limit of detection (LOD, 60 copies/mL) was observed in 5/8 infant RMs starting at 3 days post-dose 4 and peaking at 771 copies/mL. Of the 135 measurements during AZD5582 treatment in these 5 RM infants, only 8 were above the LOD (6%), lower than the 46% we have previously reported in adult RMs. Pharmacokinetic analysis of plasma AZD5582 levels revealed a lower Cmax in treated infants compared to adults (294 ng/mL versus 802 ng/mL). RNA-Sequencing of CD41 T cells comparing pre- and post-AZD5582 dosing showed many genes that were similarly upregulated in infants and adults, but the expression of key ncNF-k B genes, including NFKB2 and RELB, was significantly higher in adult RMs. Our results suggest that dosing modifications for this latency reversal approach may be necessary to maximize virus reactivation in the pediatric setting for successful “shock and kill” strategies. IMPORTANCE While antiretroviral therapy (ART) has improved HIV-1 disease outcome and reduced transmission, interruption of ART results in rapid viral rebound due to the persistent latent reservoir. Interventions to reduce the viral reservoir are of critical importance, especially for children who must adhere to lifelong ART to prevent disease progression. Here, we used our previously established pediatric nonhuman primate model of oral SIV infection to evaluate AZD5582, identified as a potent latency-reversing agent in adult macaques, in the controlled setting of daily ART. We demonstrated the safety of the IAPi AZD5582 and evaluate the pharmacokinetics and pharmacodynamics of repeated dosing. The response to AZD5582 in macaque infants differed from what we previously showed in adult macaques with weaker latency reversal in infants, likely due to altered pharmacokinetics and less inducibility of infant CD41 T cells. These data supported the contention that HIV-1 cure strategies for children are best evaluated using pediatric model systems. [ABSTRACT FROM AUTHOR]
- Published
- 2022
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21. Antiretroviral Penetration and Drug Transporter Concentrations in the Spleens of Three Preclinical Animal Models and Humans.
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Devanathan AS, Fallon JK, White NR, Schauer AP, Van Horne B, Blake K, Sykes C, Kovarova M, Adamson L, Remling-Mulder L, Luciw P, Garcia JV, Akkina R, Pirone JR, Smith PC, and Kashuba ADM
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- ATP Binding Cassette Transporter, Subfamily G, Member 2, Animals, Chromatography, Liquid, Humans, Macaca mulatta, Mice, Models, Animal, Neoplasm Proteins, Spleen, Tandem Mass Spectrometry, Anti-HIV Agents therapeutic use, HIV Infections drug therapy, Pharmaceutical Preparations
- Abstract
Adequate antiretroviral (ARV) concentrations in lymphoid tissues are critical for optimal antiretroviral therapy (ART). While the spleen contains 25% of the body's lymphocytes, there are minimal data on ARV penetration in this organ. This study quantified total and protein-unbound splenic ARV concentrations and determined whether drug transporters, sex, or infection status were modifiers of these concentrations in animal models and humans. Two humanized mice models (hu-HSC-Rag [ n = 36; 18 HIV-positive (HIV
+ ) and 18 HIV-negative (HIV- )] and bone marrow-liver-thymus [ n = 13; 7 HIV+ and 6 HIV- ]) and one nonhuman primate (NHP) model (rhesus macaque [ n = 18; 10 SHIV+ and 8 SHIV- ]) were dosed to steady state with ARV combinations. HIV+ human spleens ( n = 14) from the National NeuroAIDS Tissue Consortium were analyzed postmortem (up to 24 h postdose). ARV concentrations were measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS), drug transporter concentrations were measured with LC-MS proteomics, and protein binding in NHP spleens was determined by rapid equilibrium dialysis. Mice generally had the lowest splenic concentrations of the three species. Protein binding in splenic tissue was 6 to 96%, compared to 76 to 99% in blood plasma. NHPs had quantifiable Mrp4, Bcrp, and Ent1 concentrations, and humans had quantifiable ENT1 concentrations. None significantly correlated with tissue ARV concentrations. There was also no observable influence of infection status or sex. With these dosing strategies, NHP splenic penetration most closely resembled that of humans. These data can inform tissue pharmacokinetic scaling to humans to target HIV reservoirs by identifying important species-related differences., (Copyright © 2020 American Society for Microbiology.)- Published
- 2020
- Full Text
- View/download PDF
22. Antiretroviral Penetration across Three Preclinical Animal Models and Humans in Eight Putative HIV Viral Reservoirs.
- Author
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Devanathan AS, Pirone JR, Akkina R, Remling-Mulder L, Luciw P, Adamson L, Garcia JV, Kovarova M, White NR, Schauer AP, Blake K, Sykes C, Burgunder EM, Srinivas N, Rosen EP, and Kashuba ADM
- Subjects
- Animals, Anti-HIV Agents therapeutic use, Atazanavir Sulfate therapeutic use, Emtricitabine therapeutic use, Female, Humans, In Vitro Techniques, Maraviroc therapeutic use, Mice, Raltegravir Potassium therapeutic use, Tenofovir therapeutic use, Anti-Retroviral Agents therapeutic use, HIV Infections drug therapy
- Abstract
For HIV cure strategies like "kick and kill" to succeed, antiretroviral (ARV) drugs must reach effective concentrations in putative viral reservoirs. We characterize penetration of six ARVs in three preclinical animal models and humans. We found that standard dosing strategies in preclinical species closely mimicked tissue concentrations in humans for some, but not all, ARVs. These results have implications for interpreting HIV treatment, prevention, or cure interventions between preclinical and clinical models., (Copyright © 2019 American Society for Microbiology.)
- Published
- 2019
- Full Text
- View/download PDF
23. Antiretroviral Drug Concentrations in Lymph Nodes: A Cross-Species Comparison of the Effect of Drug Transporter Expression, Viral Infection, and Sex in Humanized Mice, Nonhuman Primates, and Humans.
- Author
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Burgunder E, Fallon JK, White N, Schauer AP, Sykes C, Remling-Mulder L, Kovarova M, Adamson L, Luciw P, Garcia JV, Akkina R, Smith PC, and Kashuba ADM
- Subjects
- Animals, Anti-HIV Agents blood, Female, HIV drug effects, Humans, Lymph Nodes drug effects, Macaca mulatta, Male, Mice, Species Specificity, Anti-HIV Agents metabolism, Anti-HIV Agents pharmacology, Gene Expression Regulation drug effects, HIV physiology, Lymph Nodes metabolism, Membrane Transport Proteins metabolism, Sex Characteristics
- Abstract
In a "kick and kill" strategy for human immunodeficiency virus (HIV) eradication, protective concentrations of antiretrovirals (ARVs) in the lymph node are important to prevent vulnerable cells from further HIV infection. However, the factors responsible for drug distribution and concentration into these tissues are largely unknown. Although humanized mice and nonhuman primates (NHPs) are crucial to HIV research, ARV tissue pharmacology has not been well characterized across species. This study investigated the influence of drug transporter expression, viral infection, and sex on ARV penetration within lymph nodes of animal models and humans. Six ARVs were dosed for 10 days in humanized mice and NHPs. Plasma and lymph nodes were collected at necropsy, 24 hours after the last dose. Human lymph node tissue and plasma from deceased patients were collected from tissue banks. ARV, active metabolite, and endogenous nucleotide concentrations were measured by liquid chromatography-tandem mass spectrometry, and drug transporter expression was measured using quantitative polymerase chain reaction and quantitative targeted absolute proteomics. In NHPs and humans, lymph node ARV concentrations were greater than or equal to plasma, and tenofovir diphosphate/deoxyadenosine triphosphate concentration ratios achieved efficacy targets in lymph nodes from all three species. There was no effect of infection or sex on ARV concentrations. Low drug transporter expression existed in lymph nodes from all species, and no predictive relationships were found between transporter gene/protein expression and ARV penetration. Overall, common preclinical models of HIV infection were well suited to predict human ARV exposure in lymph nodes, and low transporter expression suggests primarily passive drug distribution in these tissues. SIGNIFICANCE STATEMENT: During human immunodeficiency virus (HIV) eradication strategies, protective concentrations of antiretrovirals (ARVs) in the lymph node prevent vulnerable cells from further HIV infection. However, ARV tissue pharmacology has not been well characterized across preclinical species used for HIV eradication research, and the influence of drug transporters, HIV infection, and sex on ARV distribution and concentration into the lymph node is largely unknown. Here we show that two animal models of HIV infection (humanized mice and nonhuman primates) were well suited to predict human ARV exposure in lymph nodes. Additionally, we found that drug transporter expression was minimal and-along with viral infection and sex-did not affect ARV penetration into lymph nodes from any species., (Copyright © 2019 by The American Society for Pharmacology and Experimental Therapeutics.)
- Published
- 2019
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24. Predicting Efavirenz Concentrations in the Brain Tissue of HIV-Infected Individuals and Exploring their Relationship to Neurocognitive Impairment.
- Author
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Srinivas N, Joseph SB, Robertson K, Kincer LP, Menezes P, Adamson L, Schauer AP, Blake KH, White N, Sykes C, Luciw P, Eron JJ, Forrest A, Price RW, Spudich S, Swanstrom R, and Kashuba ADM
- Subjects
- Adult, Alkynes, Animals, Benzoxazines pharmacokinetics, Benzoxazines pharmacology, Cyclopropanes, Female, Humans, Macaca mulatta, Male, Middle Aged, Young Adult, Benzoxazines adverse effects, Benzoxazines therapeutic use, Brain metabolism, Cognition Disorders chemically induced, HIV Infections drug therapy
- Abstract
Sparse data exist on the penetration of antiretrovirals into brain tissue. In this work, we present a framework to use efavirenz (EFV) pharmacokinetic (PK) data in plasma, cerebrospinal fluid (CSF), and brain tissue of eight rhesus macaques to predict brain tissue concentrations in HIV-infected individuals. We then perform exposure-response analysis with the model-predicted EFV area under the concentration-time curve (AUC) and neurocognitive scores collected from a group of 24 HIV-infected participants. Adult rhesus macaques were dosed daily with 200 mg EFV (as part of a four-drug regimen) for 10 days. Plasma was collected at 8 time points over 10 days and at necropsy, whereas CSF and brain tissue were collected at necropsy. In the clinical study, data were obtained from one paired plasma and CSF sample of participants prescribed EFV, and neuropsychological test evaluations were administered across 15 domains. PK modeling was performed using ADAPT version 5.0 Biomedical Simulation Resource, Los Angeles, CA) with the iterative two-stage estimation method. An eight-compartment model best described EFV distribution across the plasma, CSF, and brain tissue of rhesus macaques and humans. Model-predicted median brain tissue concentrations in humans were 31 and 8,000 ng/mL, respectively. Model-predicted brain tissue AUC was highly correlated with plasma AUC (γ = 0.99, P < 0.001) but not CSF AUC (γ = 0.34, P = 0.1) and did not show any relationship with neurocognitive scores (γ < 0.05, P > 0.05). This analysis provides an approach to estimate PK the brain tissue in order to perform PK/pharmacodynamic analyses at the target site., (© 2019 The Authors. Clinical and Translational Science published by Wiley Periodicals, Inc. on behalf of the American Society for Clinical Pharmacology and Therapeutics.)
- Published
- 2019
- Full Text
- View/download PDF
25. Differential extracellular, but similar intracellular, disposition of two tenofovir formulations in the male genital tract.
- Author
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Dumond JB, Greene SA, Prince HMA, Chen J, Maas BM, Sykes C, Schauer AP, Blake KH, Nelson JAE, Gay CL, Kashuba ADM, and Cohen MS
- Subjects
- Adenine administration & dosage, Adenine pharmacokinetics, Anti-Retroviral Agents administration & dosage, Blood Cells cytology, Blood Cells drug effects, Emtricitabine administration & dosage, Genitalia, Male virology, Humans, Male, Reproductive Tract Infections virology, Semen cytology, Semen drug effects, Tenofovir administration & dosage, Adenine analogs & derivatives, Anti-Retroviral Agents pharmacokinetics, Emtricitabine pharmacokinetics, HIV Infections drug therapy, Leukocytes, Mononuclear drug effects, Tenofovir pharmacokinetics
- Abstract
Background: The male genital tract (MGT) is a viral sanctuary and likely HIV reservoir; understanding MGT pharmacokinetics (PK) of antiretrovirals (ARVs) used for curative strategies is critical to eradication and cure. Tenofovir alafenamide (TAF) is a tenofovir (TFV) formulation designed to maximize efficacy/minimize toxicity with unknown MGT PK., Methods: HIV-positive and HIV-negative men receiving TFV-based regimens provided six paired blood plasma (BP) and semen samples. Extracellular (TFV, TAF, emtricitabine [FTC]) drug concentrations in BP and seminal plasma (SP), and intracellular metabolite (IM) and endogenous nucleotide (EN) concentrations were measured in peripheral blood mononuclear cells (PBMCs) and seminal mononuclear cells (SMCs). Exposure ratios for SP:BP, SMC:PBMC and IM:EN were calculated from PK parameters generated by noncompartmental analysis. HIV viral load was measured in BP and SP., Results: Sixteen HIV-positive (n=8, TDF/FTC; n=8, TAF/FTC) and eight HIV-negative (TDF/FTC) men provided samples. Median TFV SP:BP ratios differed between TDF and TAF (1.5 versus 7.4), due to lower TFV BP concentrations with TAF coupled with TFV SP concentrations similar to TDF., Ftc Sp: BP ratios were approximately 3. SMC concentrations of IMs and ENs were a fraction of PBMC concentrations (1-22%), though IM:EN ratios exceed a suggested protective threshold., Conclusions: TAF SP PK was unexpected. IM SMC concentrations were low relative to PBMC, as were EN concentrations, suggesting differences in cell phenotype and lineage in the MGT; these differences in phenotype and pharmacology may have an impact on selecting and dosing ARVs used in cure strategies.
- Published
- 2019
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26. A Pharmacokinetic/Pharmacodynamic Model to Predict Effective HIV Prophylaxis Dosing Strategies for People Who Inject Drugs.
- Author
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Garrett KL, Chen J, Maas BM, Cottrell ML, Prince HA, Sykes C, Schauer AP, White N, and Dumond JB
- Subjects
- Adenine analogs & derivatives, Adenine pharmacokinetics, Adenine therapeutic use, Adult, Alanine, Emtricitabine pharmacokinetics, Emtricitabine therapeutic use, Female, Humans, Middle Aged, Pre-Exposure Prophylaxis methods, Tenofovir pharmacokinetics, Tenofovir therapeutic use, Young Adult, Anti-HIV Agents pharmacokinetics, Anti-HIV Agents therapeutic use, HIV drug effects, HIV Infections drug therapy
- Abstract
The goal of this work was to evaluate dosing strategies for tenofovir disoproxil fumarate (TDF), tenofovir alafenamide (TAF), and emtricitabine (FTC) for pre-exposure prophylaxis (PrEP) with injection drug use with a pharmacokinetic/pharmacodynamics analysis of concentration data generated from two single-dose clinical studies conducted in healthy women. Population pharmacokinetic models were developed using measured intracellular metabolite, endogenous nucleotide competitors, and extracellular parent drug concentrations. Intracellular metabolite concentrations were normalized to endogenous competitors and compared with an EC
90 target for PrEP efficacy. Monte Carlo simulations were used to select effective dose strategies of single agents (TAF, TDF, and FTC) and combinations (TDF + FTC and TAF + FTC). Daily, intermittent, and event-driven dosing regimens at varying dosage amounts were explored. When combined, TDF + FTC and TAF + FTC both provided quick (0.5 hours) and durable (up to 84 and 108 hours, respectively) protection of ≥99% after a single dose. When dosed twice per week, protection remained at 100%. Single-agent regimens provided lower estimates of protection than either combination tested. Here, the application of pharmacokinetic modeling to in vitro target concentrations demonstrates the added utility of including FTC in a successful PrEP regimen. While no TAF-based PrEP data are currently available for comparison, this analysis suggests TAF + FTC could completely protect against percutaneous exposure with as little as two doses per week., (Copyright © 2018 by The American Society for Pharmacology and Experimental Therapeutics.)- Published
- 2018
- Full Text
- View/download PDF
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