After a first dose, ZDV concentrations were detectable in both BP and GT secretions 2 h after a dose

After a first dose, ZDV concentrations were detectable in both BP and GT secretions 2 h after a dose. relative to blood plasma. Median rank order of highest to least expensive genital tract concentrations relative to blood plasma at constant state were: lamivudine (concentrations achieved were 411% greater than blood plasma), emtricitabine (395%), zidovudine (235%) tenofovir (75%), ritonavir (26%), didanosine (21%), atazanavir (18%), lopinavir (8%), abacavir (8%), stavudine (5%), and efavirenz (0.4%). Conclusions This is the first study to comprehensively evaluate antiretroviral drug exposure in the female genital tract. These findings support the use of lamivudine, zidovudine, tenofovir and emtricitabine as excellent pre-exposure/post-exposure prophylaxis (PrEP/PEP) candidates. Atazanavir and lopinavir might be useful brokers for these applications due to favorable therapeutic indices, despite lower genital tract concentrations. Brokers such as stavudine, abacavir, and efavirenz that accomplish genital tract exposures less than 10% of blood plasma are less attractive PrEP/PEP candidates. (2600 rpm) at 4C for 15 min. The producing plasma was aliquoted into labeled cryovials and stored at ?80C until analysis. Drug concentrations in BP were measured using validated Rabbit Polyclonal to MAP3K7 (phospho-Ser439) high performance liquid chromatography (HPLC)/UV methods [13C15], and concentrations in CVF were quantified using a validated HPLC-mass spectrometry (MS)/MS method [16]. Briefly, CVF concentrations were measured using a simultaneous assay for 17 antiretroviral drugs. After thawing, samples were centrifuged and the resultant supernatant underwent solid phase extraction using BOND ELUT C-18 columns (Varian, Harbor City, California, USA) as previously explained [15]. Cimetidine (in acetate buffer, pH 5.0) was used as internal standard, and was applied directly to the conditioned column prior to CVF introduction. A Shimadzu solvent delivery system (Columbia, Maryland, USA) and a LEAP HTC Pal (-)-JQ1 thermostatted (6C) autosampler (Carrboro, North Carolina, USA) connected to an Applied Biosystems API4000 triple quandruple mass spectrometer and Turbospray ion source (Applied Biosystems, Foster City, California, USA) with an Aquasil C18 column (Thermo-Electron, San Jose, California, USA) was utilized for the analysis. Multiple reaction monitoring and positive-to-negative polarity switching were used. Assay sensitivity was 1 ng/ml for abacavir (ABC); 5 ng/ml for lamivudine (3TC), zidovudine (ZDV), emtricitabine (FTC), lopinavir (LPV), atazanavir(ATV) and efavirenz (EFV); and 10 ng/ml for tenofovir (TDF), didanosine (ddI), stavudine (d4T) and ritonavir (RTV). Overall assay precision was 2.0C14.3 CV%, and accuracy was 88C113%. Recovery for the drugs analyzed ranged from 80% for RTV and LPV to 99% for 3TC, ddI and ABC. All analytical work was performed by the UNC Center for AIDS Research (CFAR) Clinical Pharmacology and Analytical Chemistry Core, which participates in quarterly national and international external proficiency screening [17,18]. These results consistently demonstrate high levels of accuracy and precision for our antiretroviral assays. Data analysis methods Pharmacokinetic parameters, including the area under the timeCconcentration curve (AUC0?), were estimated for both CVF and BP using WinNonlin (version 4.0.1, Pharsight, Inc. Mountain View, California, USA). For these computations, concentration measurements below the lower limit of detection were imputed as zero and those below the lower limit of quantitation (LLQ) were imputed as ? LLQ. For each antiretroviral agent, the GT: BP AUC0? ratios were calculated, and multiplied by 100 to represent penetration of drug into the GT relative to BP [19]. Descriptive statistical methods, particularly medians and the interquartile range (IQR), were used in the primary analyses of these AUC ratios. The 95% confidence intervals of the median AUC ratios for each drug at each visit were calculated using Intercooled STATA Release 8.0 (Stata Corporation, College Station, Texas, USA). Results Demographics Demographic data for the 27 women enrolled are offered in Table 1. The study population was predominantly African American (70%), middle-aged (median: 35 years; IQR: 31C42 years), with a BP HIV RNA of 4.7 (IQR, 4.0C5.0) log10 copies/ml and a CD4+T-cell count of 307 (220C372) cells/l at study entry. As most subjects were antiretroviral experienced (67%), the provider-selected regimens varied widely. Seventy percent (19/27) of the regimens contained 3TC. TDF was found in 56% of the regimens, ABC in 30%, and ZDV in 24%. Thirty-three percent of the.Caviness General Clinical Research Center, and the UNC CFAR Clinical Pharmacology and Analytical Chemistry Core. Sponsorship: This work was supported by the National Institute of Allergy and Infectious Diseases (AI54980; A.D.M.K.), the UNC Center for (-)-JQ1 AIDS Research/National Institute of Allergy and Infectious Disease (AI50410; N.L.R., A.S.B.), the UNC General Clinical Research Center/National Institutes of Health (RR00046), and the UNC Building Interdisciplinary Research Careers in Womens Health program (“type”:”entrez-nucleotide”,”attrs”:”text”:”HD001441″,”term_id”:”300574098″,”term_text”:”HD001441″HD001441; A.D.M.K., K.B.P.). Footnotes This work was presented in part at the 13th Conference on Retroviruses and Opportunistic Infections.. tenofovir and emtricitabine as excellent pre-exposure/post-exposure prophylaxis (PrEP/PEP) candidates. Atazanavir and lopinavir might be useful brokers for these applications due to favorable therapeutic indices, despite lower genital tract concentrations. Brokers such as stavudine, abacavir, and efavirenz that accomplish genital tract exposures less than 10% of blood plasma are less attractive PrEP/PEP candidates. (2600 rpm) at 4C for 15 min. The producing plasma was aliquoted into labeled cryovials and stored at ?80C until analysis. Drug concentrations in BP were measured using validated high performance liquid chromatography (HPLC)/UV methods [13C15], and concentrations in CVF were quantified using a validated HPLC-mass spectrometry (MS)/MS method [16]. Briefly, CVF concentrations were measured using a simultaneous assay for 17 antiretroviral drugs. After thawing, samples were centrifuged and the resultant supernatant underwent solid phase extraction using BOND ELUT C-18 columns (Varian, Harbor City, California, USA) as previously explained [15]. Cimetidine (in acetate buffer, pH 5.0) was used as internal standard, and was applied directly to the conditioned column prior to CVF introduction. A Shimadzu solvent delivery system (Columbia, Maryland, USA) and a LEAP HTC Pal thermostatted (6C) autosampler (Carrboro, North Carolina, USA) connected to an Applied Biosystems API4000 triple quandruple mass spectrometer and Turbospray ion source (Applied Biosystems, Foster City, California, USA) with an Aquasil C18 column (Thermo-Electron, San Jose, California, USA) was utilized for the analysis. Multiple reaction monitoring and positive-to-negative polarity switching were used. Assay sensitivity was 1 ng/ml for abacavir (ABC); 5 ng/ml for lamivudine (3TC), zidovudine (ZDV), emtricitabine (FTC), lopinavir (LPV), (-)-JQ1 atazanavir(ATV) and efavirenz (EFV); and 10 ng/ml for tenofovir (TDF), didanosine (ddI), stavudine (d4T) and ritonavir (RTV). Overall assay precision was 2.0C14.3 CV%, and accuracy was 88C113%. Recovery for the drugs analyzed ranged from 80% for RTV and LPV to 99% for 3TC, ddI and ABC. All analytical work was performed by the UNC Center for AIDS Research (CFAR) Clinical Pharmacology and Analytical Chemistry Core, which participates in quarterly national and international external proficiency screening [17,18]. These results consistently demonstrate high levels of accuracy and precision for our antiretroviral assays. Data analysis methods Pharmacokinetic parameters, including the area under the timeCconcentration curve (AUC0?), were estimated for both CVF and BP using WinNonlin (version 4.0.1, Pharsight, Inc. Mountain View, California, USA). For these computations, concentration measurements below the lower limit of detection were imputed as zero and those below the lower limit of quantitation (LLQ) were imputed as ? LLQ. For each antiretroviral agent, the GT: BP AUC0? ratios were calculated, and multiplied by 100 to represent penetration of drug into the GT relative to BP [19]. Descriptive statistical methods, particularly medians and the interquartile range (IQR), were used in the primary analyses of these AUC ratios. The 95% confidence intervals of the median AUC ratios for each drug at each visit were calculated using Intercooled STATA Release 8.0 (Stata Corporation, College Station, Texas, USA). Results Demographics Demographic data for the 27 women enrolled are offered in Table 1. The study population was predominantly African American (70%), middle-aged (median: 35 years; IQR: 31C42 years), with a BP HIV RNA of 4.7 (IQR, 4.0C5.0) log10 copies/ml and a CD4+T-cell count of 307 (220C372) cells/l at study entry. As most subjects were antiretroviral experienced (67%), the provider-selected regimens varied widely. Seventy percent (19/27) of the regimens contained 3TC. TDF was found in 56% of the regimens, ABC in 30%, and ZDV in 24%..