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1.
Biomed Chromatogr ; 38(7): e5888, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38727008

RESUMO

A simple and reliable HPLC-ultraviolet (HPLC-UV) method was developed and validated for the quantification of pritelivir in the samples of medium from the experiments utilizing the ex vivo technique of dual perfusion of the human placental lobule. Phenacetin was used as an internal standard (IS) in our HPLC-UV method. Chromatographic separation of pritelivir and phenacetin was achieved on a Waters Symmetry C18 HPLC column (100 × 2.1 mm, 3.5 µm) at ambient temperature (22-25°C). The mobile phase was composed of 50% methanol in deionized water (v/v), the flow rate for isocratic elution was established at 0.25 mL/min, and the detection wavelength for pritelivir and IS was set at 254 nm. Pritelivir and IS were extracted with the protein precipitation method using methanol as a solvent. The calibration curve for pritelivir exhibited linearity (r2 > 0.99) within the concentration range from 0.155 to 6.62 µg/mL. Within- and between-day accuracy ranged from 97% to 110% with relative standard deviation (RSD) values not exceeding 10%. The extraction recovery of pritelivir and IS ranged from 89% to 91% with RSD not exceeding 7%. Pritelivir was stable under the storage and sample handling conditions. This validated HPLC-UV method was utilized to quantify pritelivir in the placental perfusion medium samples, and the resulting concentrations were authenticated with incurred sample reanalysis to confirm the reliability of the method.


Assuntos
Limite de Detecção , Placenta , Cromatografia Líquida de Alta Pressão/métodos , Humanos , Placenta/química , Feminino , Gravidez , Reprodutibilidade dos Testes , Modelos Lineares , Espectrofotometria Ultravioleta/métodos , Perfusão , Sulfonamidas/análise
2.
Pharm Nanotechnol ; 2023 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-37461351

RESUMO

BACKGROUND: In this study, a prototype of a targeted nanocarrier for drug delivery for prenatal therapy of the developing fetus was developed and examined in vitro and ex vivo. The folate transport mechanism in the human placenta was utilized as a possible pathway for the transplacental delivery of targeted nanoparticles. METHODS: Several types of folic acid-decorated polymeric nanoparticles were synthesized and characterized. During transport studies of targeted and non-targeted fluorescent nanoparticles across the placental barrier, the apparent permeability values, uptake, transfer indices, and distribution in placental tissue were determined. RESULTS: The nanoparticles had no effect on BeWo b30 cell viability. In vitro, studies showed significantly higher apparent permeability of the targeted nanoparticles across the cell monolayers as compared to the nontargeted nanoparticles (Pe = 5.92 ± 1.44 ×10-6 cm/s for PLGA-PEG-FA vs. 1.26 ± 0.31 ×10-6 cm/s for PLGA-PEG, P < 0.05), and the transport of the targeted nanoparticles was significantly inhibited by excess folate. Ex vivo placental perfusion showed significantly greater accumulation of the targeted nanoparticles in the placental tissue (4.31 ± 0.91%/g for PLGA-PEG-FA vs. 2.07 ± 0.26%/g for PLGA-PEG). CONCLUSION: The data obtained suggested different mechanisms for the uptake and transplacental transfer of targeted versus nontargeted nanoparticles. This targeted nanoformulation may be a promising strategy for fetal drug therapy.

3.
Drug Metab Pharmacokinet ; 47: 100477, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36368298

RESUMO

Although methadone is effective in the management of acute pain, the complexity of its absorption-distribution-metabolism-excretion profile limits its use as an opioid of choice for perioperative analgesia. Because deuteration is known to improve the pharmacokinetic, pharmacodynamic and toxicological properties of some drugs, here we characterized the single dose pharmacokinetic properties and post-operative analgesic efficacy of d9-methadone. The pharmacokinetic profiles of d9-methadone and methadone administered intravenously to CD-1 male mice revealed that deuteration leads to a 5.7- and 4.4-fold increase in the area under the time-concentration curve and maximum concentration in plasma, respectively, as well as reduction in clearance (0.9 ± 0.3 L/h/kg vs 4.7 ± 0.8 L/h/kg). The lower brain-to-plasma ratio of d9-methadone compared to that of methadone (0.35 ± 0.12 vs 2.05 ± 0.62) suggested that deuteration decreases the transfer of the drug across the blood-brain barrier. The estimated LD50 value for a single intravenous dose of d9-methadone was 2.1-fold higher than that for methadone. Moreover, d9-methadone outperformed methadone in the efficacy against postoperative pain by primarily activating peripheral opioid receptors. Collectively, these data suggest that the replacement of three hydrogen atoms in three methyl groups of methadone altered its pharmacokinetic properties, improved safety, and enhanced its analgesic efficacy.


Assuntos
Analgésicos Opioides , Metadona , Masculino , Animais , Camundongos , Metadona/farmacologia , Metadona/uso terapêutico , Analgésicos Opioides/farmacologia , Barreira Hematoencefálica , Cinética , Encéfalo
4.
J Biomed Nanotechnol ; 18(2): 589-599, 2022 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-35484740

RESUMO

The purpose of this study was to conduct initial characterization of membrane vesicles isolated from human placenta by agitation of villous tissue (apical and basal) as well as vesicles obtained following dual perfusion of placental lobule. The morphology, physical and biological properties of the isolated vesicles were determined by electron microscopy, dynamic light scattering, and immunoblotting as well as nanoflow liquid chromatography-mass spectrometry proteomics analysis. CD-1 male mice were used to test the biocompatibility of the vesicles in vivo and assess the biodistribution of fluorescently labeled apical and perfusion vesicles. The vesicles obtained following placental perfusion and the apical vesicles had Z-average diameters of 199±23 nm and 246±24 nm, respectively, and demonstrated nanocarrier stability, low toxicity, and low immunogenicity. On the other hand, administration of basal vesicles resulted in animal demise with LD50 of 0.85 µgprotein/g. Both fluorescently labeled apical and perfusion vesicles were detected in the lungs, liver, kidneys, and spleen of CD-1 mice within 24 h of administration. However, there were differences in organ distribution of these vesicles over 24 hours time period. These data suggest that placental apical and perfusion vesicles have a potential for further development as biological vehicles for drug delivery.


Assuntos
Sistemas de Liberação de Medicamentos , Placenta , Animais , Feminino , Humanos , Fígado , Pulmão , Masculino , Camundongos , Placenta/metabolismo , Gravidez , Distribuição Tecidual
5.
J Pharm Sci ; 111(2): 505-516, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-34597623

RESUMO

Pravastatin is currently under evaluation for prevention of preeclampsia. Factors contributing to placental disposition of pravastatin are important in assessment of potential undesirable fetal effects. The purpose of this study was to identify the uptake transporters that contribute to the placental disposition of pravastatin. Our data revealed the expression of organic anion transporting polypeptide 1A2 (OATP1A2) and OATP2A1 in the apical, and OATP2B1 and OATP5A1 in the basolateral membranes of the placenta, while organic anion transporter 4 (OAT4) exhibited higher expression in basolateral membrane but was detected in both membranes. Preloading placental membrane vesicles with glutarate increased the uptake of pravastatin suggesting involvement of glutarate-dependent transporters such as OAT4. In the HEK293 cells overexpressing individual uptake transporters, OATP2A1, OATP1A2 and OAT4 were determined to accept pravastatin as a substrate at physiological pH, while the uptake of pravastatin by OATP2B1 (known to interact with pravastatin at acidic pH) and OATP5A1 was not detected at pH 7.4. These findings led us to propose that OATP1A2 and OATP2A1 are responsible for the placental uptake of pravastatin from the maternal circulation, while OAT4 mediates the passage of the drug across placental basolateral membrane in the fetal-to-maternal direction.


Assuntos
Transportadores de Ânions Orgânicos/metabolismo , Pravastatina , Transporte Biológico , Feminino , Células HEK293 , Humanos , Placenta/metabolismo , Pravastatina/metabolismo , Gravidez
6.
Nanomedicine ; 33: 102354, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33429062

RESUMO

Diagnosis and treatment of breast cancer in pregnancy can result in morbidity and mortality for the mother and fetus. Many new paclitaxel nanoformulations commercially available worldwide for breast cancer treatment are being adopted due to favorable dosing regimens and side effect profiles, but their transplacental transport and resultant fetal exposure remain unknown. Here, we examine three formulations: Taxol (paclitaxel dissolved in Kolliphor EL and ethanol); Abraxane (albumin nanoparticle); and Genexol-PM (polymeric micelle). In the ex vivo dually perfused human placental cotyledon, placental accumulation of Genexol-PM is higher than Taxol, and both nanoformulations have lower maternal concentrations of paclitaxel over time. In vitro studies of these formulations and fluorescent nanoparticle analogs demonstrate that Genexol-PM allows paclitaxel to overcome P-glycoprotein efflux, but Abraxane behaves as a free drug formulation. We anticipate that these findings will impact future development of rational and safe treatment strategies for pregnancy-associated breast cancer and other diseases.


Assuntos
Antineoplásicos Fitogênicos/química , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/tratamento farmacológico , Portadores de Fármacos/química , Paclitaxel/química , Paclitaxel Ligado a Albumina/química , Albuminas/química , Linhagem Celular Tumoral , Composição de Medicamentos , Feminino , Humanos , Micelas , Nanopartículas/química , Paclitaxel/farmacologia , Placenta/citologia , Polietilenoglicóis/química , Gravidez
7.
Am J Obstet Gynecol ; 220(5): 498.e1-498.e9, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30849355

RESUMO

BACKGROUND: The transfer of pathogenic immunoglobulin G antibodies from mother to fetus is a critical step in the pathophysiology of alloimmune and autoimmune diseases of the fetus and neonate. Immunoglobulin G transfer across the human placenta to the fetus is mediated by the neonatal Fc receptor, and blockade of the neonatal Fc receptor may provide a therapeutic strategy to prevent or minimize pathological events associated with immune-mediated diseases of pregnancy. M281 is a fully human, aglycosylated monoclonal immunoglobulin G1 antineonatal Fc receptor antibody that has been shown to block the neonatal Fc receptor with high affinity in nonclinical studies and in a phase 1 study in healthy volunteers. OBJECTIVE: The objective of the study was to determine the transplacental transfer of M281 and its potential to inhibit transfer of immunoglobulin G from maternal to fetal circulation. STUDY DESIGN: To determine the concentration of M281 required for rapid cellular uptake and complete saturation of the neonatal Fc receptor in placental trophoblasts, primary human villous trophoblasts were incubated with various concentrations of M281 in a receptor occupancy assay. The placental transfer of M281, immunoglobulin G, and immunoglobulin G in the presence of M281 was studied using the dually perfused human placental lobule model. Immunoglobulin G transfer was established using a representative immunoglobulin G molecule, adalimumab, a human immunoglobulin G1 monoclonal antibody, at a concentration of 270 µg/mL. Inhibition of immunoglobulin G transfer by M281 was determined by cotransfusing 270 µg/mL of adalimumab with 10 µg/mL or 300 µg/mL of M281. Concentrations of adalimumab and M281 in sample aliquots from maternal and fetal circuits were analyzed using a sandwich enzyme-linked immunosorbent assay and Meso Scale Discovery assay, respectively. RESULTS: In primary human villous trophoblasts, the saturation of the neonatal Fc receptor by M281 was observed within 30-60 minutes at 0.15-5.0 µg/mL, suggesting rapid blockade of neonatal Fc receptor in placental cells. The transfer rate of adalimumab (0.23% ± 0.21%) across dually perfused human placental lobule was significantly decreased by 10 µg/mL and 300 µg/mL of M281 to 0.07 ± 0.01% and 0.06 ± 0.01%, respectively. Furthermore, the transfer rate of M281 was 0.002% ± 0.02%, approximately 100-fold lower than that of adalimumab. CONCLUSION: The significant inhibition of immunoglobulin G transfer across the human placental lobule by M281 and the minimal transfer of M281 supports the development of M281 as a novel agent for the treatment of fetal and neonatal diseases caused by transplacental transfer of alloimmune and autoimmune pathogenic immunoglobulin G antibodies.


Assuntos
Anticorpos Monoclonais/farmacologia , Imunoglobulina G/metabolismo , Troca Materno-Fetal/imunologia , Placenta/imunologia , Receptores Fc/imunologia , Adalimumab , Transporte Biológico , Feminino , Humanos , Imunoglobulina G/imunologia , Modelos Biológicos , Placenta/metabolismo , Gravidez , Trofoblastos/imunologia
8.
Biochem Pharmacol ; 156: 467-478, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-30217571

RESUMO

The expression and activity of human placental transporters during pregnancy could be altered by several factors including pathological changes associated with preeclampsia. The aims of this study were to identify the placental efflux transporters involved in the bio-disposition of pravastatin, determine the protein expression of these transporters and their encoding genes as well as the activity of pravastatin uptake in placentas obtained from patients with preeclampsia. ATP-dependent uptake of [3H]-pravastatin by trophoblast tissue apical and basal membrane vesicles exhibited sigmoidal kinetics. The curved shapes of Eadie-Hofstee plots indicate that more than one placental transporter are involved in the uptake of pravastatin. ATP-dependent uptake of [3H]-pravastatin into vesicles expressing MRP1-5, BCRP, and P-gp, as well as the results of inhibition studies suggest that BCRP and MRP1 are the major placental efflux transporters responsible for the in vitro uptake of pravastatin. Compared to placentas from healthy pregnancies, preeclamptic placentas had increased number of syncytial knots with increased expression of BCRP in their apical membrane and increased expression of MRP1 in the cytoplasm of the syncytiotrophoblast and in cytoplasm of syncytial knots. There was a concomitant increase in ABCC1 but not in ABCG2 gene expressions in preeclamptic placentas. ATP-dependent uptake of [3H]-pravastatin by vesicles prepared from apical membranes of preeclamptic placentas was similar to the uptake by vesicles prepared from placentas obtained after uncomplicated pregnancies (13.9 ±â€¯6.5 vs 14.1 ±â€¯5.8 pmol·mg protein-1 min-1). The transporter-specific changes in the expression of BCRP and MRP1 in preeclamptic placentas did not affect the efflux activity of transporters localized on the apical membrane of the syncytiotrophoblast.


Assuntos
Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/metabolismo , Proteínas Associadas à Resistência a Múltiplos Medicamentos/metabolismo , Proteínas de Neoplasias/metabolismo , Placenta/metabolismo , Pravastatina/metabolismo , Transporte Biológico , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Pré-Eclâmpsia/metabolismo , Gravidez
9.
J Liq Chromatogr Relat Technol ; 41(5): 232-238, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30774296

RESUMO

Ex vivo placental perfusion experiments are important in understanding the quantity and mechanisms of xenobiotic transport to the fetus during pregnancy. Our study demonstrates that paclitaxel and antipyrine concentrations in placental perfusion medium containing physiological concentrations of human serum albumin during pregnancy (30 mg/mL) can be quantified by RP-HPLC and UV detection. A liquid-liquid extraction method was developed for the quantification of paclitaxel and celecoxib (internal standard) from perfusion medium. Antipyrine, which is a necessary marker in placental perfusions for determining the validity of experiments and calculating the clearance index of xenobiotics, was also analyzed by HPLC and UV detection. Antipyrine concentrations were determined by HPLC after precipitating the perfusion medium in acetonitrile and separating the precipitated proteins by centrifugation. Concentrations were fitted to linear regressions with R2 values approaching 1. Lower limits of detection for paclitaxel and antipyrine were 100 ng/mL and 200 ng/mL, respectively. Both methods demonstrated high intra-day and inter-day precision and trueness. Additionally, the use of these methods was demonstrated in a placental perfusion experiment using Taxol® (paclitaxel dissolved in Cremophor-EL). The fetal transfer rate of Taxol was 6.6% after 1 hour.

10.
Drug Metab Lett ; 11(1): 29-34, 2017 11 17.
Artigo em Inglês | MEDLINE | ID: mdl-28595558

RESUMO

BACKGROUND: Bupropion (BUP) has a potential to be an effective pharmacotherapy for smoking cessation during pregnancy. Smoking during pregnancy stimulates placental carbonyl reductases that catalyze the biotransformation of BUP. 4-methylnitrosamino-1-(3-pyridyl)-1-butanone (NNK) is a potent carcinogen of cigarette smoke. Carbonyl reduction of NNK into 4- methylnitrosamino-1-(3-pyridyl)-1-butanol (NNAL) constitutes a major step in NNK detoxification. Thus, placentas of pregnant smokers on BUP therapy can become a site of drug-drug interaction. Therefore, we investigated the effect of continuous exposure to BUP and cigarette smoke on the activity of placental carbonyl reductases in the formation of NNAL from NNK. METHODS: The reductive metabolism of NNK was determined using microsomal and cytosolic subcellular fractions of placentas obtained from non-smoking women treated with BUP for depression, and women not exposed to BUP: non-smokers (control) and smokers. The effect of BUP and its metabolites on the reductive metabolism of NNK was investigated using subcellular fractions of control placentas. RESULTS: The formation of NNAL from NNK by placental cytosolic fractions of heavy smokers (≥20 cigarettes per day) was lower than that of control (12.1±3.5 nmol.mgP-1 vs 16.5±6.0 nmol.mgP-1, P<0.05). While being exposed to BUP, the activity of placental carbonyl reductases remained unaffected, the formation of NNAL in the placental cytosolic fraction decreased only in the presence of high concentrations of BUP metabolites. CONCLUSION: Smoking during pregnancy decreases the detoxifying capacity of soluble carbonyl reductases towards NNK. Given the experimental conditions, exposure to BUP and its metabolites should not impede the reductive metabolism of NNK by placenta in vivo.


Assuntos
Antidepressivos/metabolismo , Bupropiona/metabolismo , Nitrosaminas/metabolismo , Placenta/metabolismo , Antidepressivos/farmacocinética , Biotransformação , Bupropiona/farmacocinética , Fumar Cigarros/metabolismo , Feminino , Humanos , Microssomos/metabolismo , Placenta/enzimologia , Gravidez , Frações Subcelulares/metabolismo , Trofoblastos/metabolismo
11.
Am J Obstet Gynecol ; 209(4): 373.e1-5, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24070396

RESUMO

OBJECTIVE: The objective of the study was to determine the bidirectional transfer of pravastatin across the dually perfused term human placental lobule and its distribution between the tissue and maternal and fetal circuits. STUDY DESIGN: The transfer of pravastatin was determined in the maternal-to-fetal (n = 11) and fetal-to-maternal (n = 10) directions. Pravastatin was coperfused with its [(3)H]-isotope and the marker compound antipyrine (20 µg/mL) and its [(14)C]-isotope. The concentration of pravastatin in the perfused tissue and the maternal and fetal circuits was determined using liquid scintillation spectrometry. Inside-out vesicles prepared from placental brush border membranes were utilized to investigate the role of efflux transporters in the transplacental transfer of pravastatin. RESULTS: Pravastatin was transferred from the maternal to the fetal circuit and vice versa. In the maternal-to-fetal direction, the distribution of pravastatin at the end of experiment was as follows: 14 ± 5% of the drug was retained by the tissue, 68 ± 5% remained in the maternal circuit, and 18 ± 4% was transferred to the fetal circuit. The normalized transfer of pravastatin (clearance index) to antipyrine in the fetal-to-maternal direction (0.48 ± 0.07) was higher than its transfer in the maternal-to-fetal direction (0.36 ± 0.07, P < .01). Furthermore, pravastatin inhibited the adenosine triphosphate (ATP)-dependent uptake of the paclitaxel and estrone sulfate. CONCLUSION: The transfer of pravastatin across the dually perfused placental lobule suggests that fetal exposure to pravastatin is plausible. The higher transfer of pravastatin in the fetal-to-maternal direction than the reverse as well as its inhibition of the ATP-dependent uptake of [(3)H]-paclitaxel and [(3)H]-estrone sulfate strongly suggest the involvement of efflux transporters in decreasing its transfer across the placenta and support pravastatin's favorable pharmacokinetic profile in pregnancy.


Assuntos
Inibidores de Hidroximetilglutaril-CoA Redutases/farmacocinética , Troca Materno-Fetal , Placenta/metabolismo , Pravastatina/farmacocinética , Feminino , Humanos , Gravidez
12.
Am J Obstet Gynecol ; 207(4): 331.e1-6, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22867688

RESUMO

OBJECTIVE: We sought to determine the bidirectional transfer and distribution of vancomycin and telavancin across the dually perfused term human placental lobule. STUDY DESIGN: The technique of dually perfused placental lobule was used in its recirculating mode to determine the maternal to fetal (M→F) (n = 20) and fetal to maternal (n = 18) transfer of each antibiotic, which were coperfused with their radioactive isotopes. The concentrations of drugs were determined by liquid scintillation spectrometry. RESULTS: In the M→F direction, the transfer of vancomycin (9.6 ± 4%) and telavancin (6.5 ± 2%) were low; however, telavancin retention by the perfused lobule was greater than that of vancomycin (P < .01). The normalized transplacental transfer of telavancin across the placental lobule in the fetal to maternal direction was higher than in the M→F direction (P < .01), suggesting the involvement of placental efflux transporters. CONCLUSION: The ex vivo perfusion experiments revealed low transfer of vancomycin and telavancin to the fetal circuit.


Assuntos
Aminoglicosídeos/farmacocinética , Antibacterianos/farmacocinética , Troca Materno-Fetal , Placenta/metabolismo , Vancomicina/farmacocinética , Feminino , Humanos , Lipoglicopeptídeos , Perfusão , Placenta/efeitos dos fármacos , Gravidez
13.
J Matern Fetal Neonatal Med ; 25(11): 2312-5, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22590979

RESUMO

OBJECTIVES: Determine the bidirectional transfer of oseltamivir carboxylate (OC) across term human placenta and its distribution between the tissue, maternal and fetal circuits. METHODS: The technique of dual perfusion of placental lobule (DPPL) in its recirculating mode was utilized to determine the transfer of the drug. OC (350 ng/mL) was co-perfused with its [(3)H]-isotope and the marker compound antipyrine (AP, 20 µg/mL) together with its [(14)C]-isotope. The concentrations of OC and any of its metabolite(s) formed during perfusion were determined in the tissue, maternal and fetal circuits by liquid scintillation spectrometry following their separation by High Performance Liquid Chromatography (HPLC). RESULTS: The distribution of OC following its perfusion in the Maternal-to-Fetal direction for 4 h was as follows: 21 ± 4% of the drug was transferred to the fetal circuit, 13 ± 5% was retained by the perfused lobule, and 66 ± 4% remained in the maternal circuit. The normalized transfer of OC to that of AP (Clearance index) in the maternal-to-fetal direction was (0.47 ± 0.11) and was not different from its transfer from the fetal-to-maternal direction (0.47 ± 0.06) suggesting that involvement of placental efflux transporters is unlikely. CONCLUSIONS: OC crosses human placenta. As the transfer rate of OC is 47% of the freely diffusible AP, it is likely that fetus could be exposed to OC during pregnancy.


Assuntos
Oseltamivir/análogos & derivados , Placenta/metabolismo , Antivirais/farmacocinética , Transporte Biológico/fisiologia , Células Cultivadas , Difusão , Feminino , Humanos , Troca Materno-Fetal/fisiologia , Técnicas de Cultura de Órgãos , Oseltamivir/farmacocinética , Perfusão , Gravidez , Nascimento a Termo/metabolismo , Distribuição Tecidual
14.
Am J Perinatol ; 28(1): 25-32, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20607647

RESUMO

We sought to determine whether gestational age affects the transplacental transfer and metabolism of buprenorphine (BUP). Transfer of BUP (10 ng/mL) and its [ (3)H]-isotope was determined across placentas of 30 to 34 weeks of gestation utilizing the technique of dual perfusion of placental lobule. Concentration of the drug in trophoblast tissue and in maternal and fetal circuits was determined by liquid scintillation spectrometry. Microsomes prepared from placentas of 17 to 37 weeks of gestation were divided into three groups: late second, early third, and late third trimesters. Antibodies raised against human cytochrome P450 (CYP) isoforms were utilized to identify the enzyme(s) catalyzing BUP biotransformation by preterm placental microsomes. The amount of norbuprenorphine formed was determined by liquid chromatography-mass spectrometry (LC-MS). BUP transfer across the placentas of 30 to 34 weeks of gestation was similar to those at term. CYP19 antibodies caused 60% inhibition of BUP metabolism by microsomes of late second and early third trimesters and 85% by microsomes of late third trimester. The developmental changes occurring in human placenta between 30 weeks of gestation through term do not affect the transfer of BUP across human placenta. CYP19 is the major enzyme responsible for biotransformation of BUP beginning at 17 weeks of gestation until term.


Assuntos
Buprenorfina/análogos & derivados , Buprenorfina/farmacocinética , Sistema Enzimático do Citocromo P-450/metabolismo , Microssomos/enzimologia , Placenta/enzimologia , Anticorpos Monoclonais , Aromatase/imunologia , Aromatase/metabolismo , Hidrocarboneto de Aril Hidroxilases/imunologia , Hidrocarboneto de Aril Hidroxilases/metabolismo , Biotransformação , Buprenorfina/metabolismo , Citocromo P-450 CYP2B6 , Citocromo P-450 CYP2C8 , Feminino , Idade Gestacional , Humanos , Técnicas In Vitro , Oxirredutases N-Desmetilantes/imunologia , Oxirredutases N-Desmetilantes/metabolismo , Perfusão , Placenta/fisiologia , Gravidez
15.
Biochem Pharmacol ; 80(7): 1080-6, 2010 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-20599802

RESUMO

Cigarette smoking during pregnancy is a preventable risk factor associated with maternal and fetal complications. Bupropion is an antidepressant used successfully for smoking cessation in non-pregnant patients. Our goal is to determine whether it could benefit the pregnant patient seeking smoking cessation. The aim of this investigation was to determine the role of human placenta in the disposition of bupropion and its major hepatic metabolite, OH-bupropion. The expression of efflux transporters P-gp and BCRP was determined in placental brush border membrane (n=200) and revealed a positive correlation (p<0.05). Bupropion was transported by BCRP (K(t) 3 microM, V(max) 30 pmol/mg protein/min) and P-gp (K(t) 0.5 microM, V(max) 6 pmol/mg protein min) in placental inside-out vesicles (IOVs). OH-bupropion crossed the dually-perfused human placental lobule without undergoing further metabolism, nor was it an efflux substrate of P-gp or BCRP. In conclusion, our data indicate that human placenta actively regulates the disposition of bupropion (via metabolism, active transport), but not its major hepatic metabolite, OH-bupropion.


Assuntos
Antidepressivos/metabolismo , Bupropiona/metabolismo , Placenta/metabolismo , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Transporte Biológico/efeitos dos fármacos , Transporte Biológico/fisiologia , Feminino , Humanos , Proteínas de Membrana Transportadoras/metabolismo , Gravidez
16.
Am J Obstet Gynecol ; 202(4): 383.e1-7, 2010 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-20350646

RESUMO

OBJECTIVE: Substrates of placental efflux transporters could compete for a single transporter, which could result in an increase in the transfer of each substrate to the fetal circulation. Our aim was to determine the role of placental transporters in the biodisposition of oral hypoglycemic drugs that could be used as monotherapy or in combination therapy for gestational diabetes. STUDY DESIGN: Inside-out brush border membrane vesicles from term placentas were used to determine the efflux of glyburide, rosiglitazone, and metformin by P-glycoprotein, breast cancer resistance protein, and multidrug resistance protein. RESULTS: Glyburide was transported by multidrug resistance protein (43 +/- 4%); breast cancer resistance protein (25 +/- 5%); and P-glycoprotein (9 +/- 5%). Rosiglitazone was transported predominantly by P-glycoprotein (71 +/- 26%). Metformin was transported by P-glycoprotein (58 +/- 20%) and breast cancer resistance protein (25 +/- 14%). CONCLUSION: Multiple placental transporters contribute to efflux of glyburide, rosiglitazone, and metformin. Administration of drug combinations could lead to their competition for efflux transporters.


Assuntos
Glibureto/farmacocinética , Hipoglicemiantes/farmacocinética , Metformina/farmacocinética , Placenta/metabolismo , Tiazolidinedionas/farmacocinética , Trifosfato de Adenosina/metabolismo , Transporte Biológico/fisiologia , Radioisótopos de Carbono , Proteínas de Transporte/metabolismo , Polaridade Celular/fisiologia , Diabetes Gestacional/tratamento farmacológico , Diabetes Gestacional/metabolismo , Interações Medicamentosas , Feminino , Humanos , Troca Materno-Fetal , Microvilosidades/metabolismo , Gravidez , Rosiglitazona , Trítio
17.
Biochem Pharmacol ; 79(11): 1684-90, 2010 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-20109440

RESUMO

Smoking during pregnancy is the largest modifiable risk factor for pregnancy-related morbidity and mortality. The success of bupropion for smoking cessation warrants its investigation for the treatment of pregnant patients. Nevertheless, the use of bupropion for the treatment of pregnant smokers requires additional data on its bio-disposition during pregnancy. Therefore, the aim of this investigation was to determine the metabolism of bupropion in placentas obtained from nonsmoking and smoking women, identify metabolites formed and the enzymes catalyzing their formation, as well as the kinetics of the reaction. Data obtained revealed that human placentas metabolized bupropion to hydroxybupropion, erythro- and threohydrobupropion. The rates for formation of erythro- and threohydrobupropion exceeded that for hydroxybupropion by several folds, were dependent on the concentration of bupropion and exhibited saturation kinetics with an apparent K(m) value of 40microM. Human placental 11beta-hydroxysteroid dehydrogenases were identified as the major carbonyl-reducing enzymes responsible for the reduction of bupropion to threo- and erythrohydrobupropion in microsomal fractions. On the other hand, CYP2B6 was responsible for the formation of OH-bupropion. These data suggest that both placental microsomal carbonyl-reducing and oxidizing enzymes are involved in the metabolism of bupropion.


Assuntos
Bupropiona/metabolismo , Placenta/metabolismo , Fumar/tratamento farmacológico , 11-beta-Hidroxiesteroide Desidrogenases/metabolismo , Antidepressivos de Segunda Geração , Hidrocarboneto de Aril Hidroxilases/metabolismo , Bupropiona/farmacocinética , Células Cultivadas , Citocromo P-450 CYP2B6 , Inibidores da Captação de Dopamina , Feminino , Humanos , Microssomos/enzimologia , Oxirredutases N-Desmetilantes/metabolismo , Placenta/enzimologia , Gravidez/metabolismo
18.
J Matern Fetal Neonatal Med ; 23(5): 409-16, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-19658039

RESUMO

OBJECTIVE: In order to evaluate the potential use of bupropion as smoking cessation therapy during pregnancy, the aim of this investigation was to determine transplacental transfer and metabolism of bupropion and its distribution among placental tissue and maternal and fetal circuits of the dually perfused placental lobule. METHODS: Placentas obtained from healthy term pregnancies were perfused with bupropion at two concentrations 150 ng/ml and 450 ng/ml, along with the marker compound antipyrine 20 microg/ml. Radioactive isotopes of the two drugs were co-transfused to enhance their detection limits. Concentrations of bupropion and its metabolite were determined by liquid chromatography and liquid scintillation spectrometry. RESULTS: The fetal/maternal concentration ratio of bupropion was 1.07 +/- 0.22. Following 4 h of its perfusion, 48 +/- 6% of bupropion was retained by placental tissue, 32 +/- 5% remained in the maternal circuit, and 20 +/- 6% was transferred to the fetal circuit. A metabolite of bupropion, threohydrobupropion, was identified. CONCLUSIONS: Bupropion was transferred from the maternal to fetal circuit and was biotransformed by placental tissue enzymes to its metabolite threohydrobupropion. Bupropion and its metabolite did not affect placental tissue viability or functional parameters. These data suggest that bupropion has the potential of being used for smoking cessation during pregnancy and should be further investigated for its safety and efficacy.


Assuntos
Bupropiona/metabolismo , Bupropiona/farmacocinética , Placenta/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Gonadotropina Coriônica/análise , Gonadotropina Coriônica/sangue , Inibidores da Captação de Dopamina/metabolismo , Inibidores da Captação de Dopamina/farmacocinética , Inibidores da Captação de Dopamina/uso terapêutico , Feminino , Humanos , Troca Materno-Fetal/efeitos dos fármacos , Modelos Biológicos , Técnicas de Cultura de Órgãos , Perfusão , Placenta/citologia , Placenta/efeitos dos fármacos , Gravidez , Ligação Proteica , Albumina Sérica/metabolismo , Abandono do Hábito de Fumar/métodos , Cordão Umbilical/metabolismo
19.
Biochem Pharmacol ; 79(6): 921-5, 2010 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-19896927

RESUMO

The ABC transporter P-glycoprotein is a product of the MDR1 gene and its function in human placenta is to extrude xenobiotics from the tissue thus decreasing fetal exposure. The goal of this investigation was to examine the effect of three polymorphisms in the MDR1 gene on the expression and activity of placental P-gp. In 199 term placentas examined, the C1236T variant was associated with 11% lower P-gp protein expression than wild-type, while the C3435T and G2677T/A variants each were associated with a 16% reduction (p<0.05). Homozygotes for the C1236T and C3435T variant allele (TT) were associated with 42% and 47% increase in placental P-gp transport activity, respectively (p=0.04 and p=0.02) of the prototypic substrate, [(3)H]-paclitaxel. These findings indicate that the C3435T and G2677T/A SNPs in MDR1 are significantly associated with decreased placental P-gp protein expression, while the C1236T and C3245T homozygous variants are significantly associated with an increase in its efflux activity.


Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/genética , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Regulação da Expressão Gênica/fisiologia , Placenta/metabolismo , Polimorfismo Genético , Alelos , Transporte Biológico , Feminino , Genótipo , Humanos , Gravidez
20.
Biochem Pharmacol ; 78(9): 1272-8, 2009 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-19591810

RESUMO

The use of either methadone or buprenorphine for treatment of the pregnant opiate-dependent patient improves maternal and neonatal outcome. However, patient outcomes are often complicated by neonatal abstinence syndrome (NAS). The incidence and severity of NAS should depend on opiate concentration in the fetal circulation. Efflux transporters expressed in human placental brush border membranes decrease fetal exposure to medications by their extrusion to the maternal circulation. Accordingly, the concentration of either methadone or buprenorphine in the fetal circulation is, in part, dependent on the activity of the efflux transporters. The objective of this study was to characterize the activity of P-gp and its interaction with opiates in the placental apical membrane. Therefore, brush border membrane vesicles were prepared from human placenta. The vesicles were oriented approximately 75% inside-out, exhibited saturable ATP-dependent uptake of P-gp substrate [(3)H]-paclitaxel with an apparent K(t) of 66+/-38 nM and V(max) of 20+/-3 pmol mg protein (-1)min(-1). Methadone, buprenorphine, and morphine inhibited paclitaxel transport with apparent K(i) of 18, 44, and 90 microM, respectively. Our data indicate that a method has been established to determine the activity of the efflux transporter P-gp, expressed in placental brush border membranes, and the kinetics for the transfer of its prototypic substrate paclitaxel. Furthermore, the method was used to determine the effects of methadone, buprenorphine, and morphine on paclitaxel transfer by placental P-gp and revealed that they have higher affinity to the transporter than its classical inhibitor verapamil (K(i), 300 microM).


Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Analgésicos Opioides/farmacologia , Antineoplásicos Fitogênicos/farmacocinética , Paclitaxel/farmacocinética , Placenta/metabolismo , Western Blotting , Cromatografia de Afinidade , Feminino , Humanos , Recém-Nascido , Microscopia Eletrônica , Síndrome de Abstinência Neonatal/etiologia , Gravidez
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