RESUMO
BACKGROUND: The human ether-a-go-go related gene 1 (hERG1), which codes for a potassium ion channel, is a key element in the cardiac delayed rectified potassium current, IKr, and plays an important role in the normal repolarization of the heart's action potential. Many approved drugs have been withdrawn from the market due to their prolongation of the QT interval. Most of these drugs have high potencies for their principal targets and are often irreplaceable, thus "rehabilitation" studies for decreasing their high hERG1 blocking affinities, while keeping them active at the binding sites of their targets, have been proposed to enable these drugs to re-enter the market. METHODS: In this proof-of-principle study, we focus on cisapride, a gastroprokinetic agent withdrawn from the market due to its high hERG1 blocking affinity. Here we tested an a priori strategy to predict a compound's cardiotoxicity using de novo drug design with molecular docking and Molecular Dynamics (MD) simulations to generate a strategy for the rehabilitation of cisapride. RESULTS: We focused on two key receptors, a target interaction with the (adenosine) receptor and an off-target interaction with hERG1 channels. An analysis of the fragment interactions of cisapride at human A2A adenosine receptors and hERG1 central cavities helped us to identify the key chemical groups responsible for the drug activity and hERG1 blockade. A set of cisapride derivatives with reduced cardiotoxicity was then proposed using an in-silico two-tier approach. This set was compared against a large dataset of commercially available cisapride analogs and derivatives. CONCLUSIONS: An interaction decomposition of cisapride and cisapride derivatives allowed for the identification of key active scaffolds and functional groups that may be responsible for the unwanted blockade of hERG1.
Assuntos
Cisaprida/análogos & derivados , Cisaprida/farmacologia , Canais de Potássio Éter-A-Go-Go/metabolismo , Fármacos Gastrointestinais/farmacologia , Receptor A2A de Adenosina/metabolismo , Cisaprida/efeitos adversos , Cisaprida/química , Desenho de Fármacos , Canal de Potássio ERG1 , Fármacos Gastrointestinais/efeitos adversos , Fármacos Gastrointestinais/química , Humanos , Síndrome do QT Longo/induzido quimicamente , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Receptores 5-HT4 de Serotonina/metabolismoRESUMO
Novel compounds were prepared that united the pharmacologies of the mu-opioid tramadol with the 5-HT4 agonists metoclopramide and norcisapride. The synthesis, chiral separation and in vitro activity of the new compounds is described.
Assuntos
Analgésicos Opioides/química , Receptores Opioides mu/agonistas , Agonistas do Receptor 5-HT4 de Serotonina , Analgésicos Opioides/efeitos adversos , Analgésicos Opioides/farmacologia , Cisaprida/análogos & derivados , Cisaprida/química , Cisaprida/farmacologia , Trânsito Gastrointestinal/efeitos dos fármacos , Humanos , Metoclopramida/química , Metoclopramida/farmacologia , Receptores Opioides mu/metabolismo , Receptores 5-HT4 de Serotonina/metabolismo , Sistema Respiratório/efeitos dos fármacos , Estereoisomerismo , Tramadol/químicaRESUMO
BACKGROUND: Cisapride, a prokinetic agent and substrate for cytochrome P450 (CYP) 3A4, has been used to treat neonates and infants with feeding intolerance and apnea or bradycardia associated with gastroesophageal reflux. At age 1 month, CYP3A4 activity has been reported to be only 30% to 40% of adult activity. This known developmental delay in the expression of CYP3A4 prompted us to conduct a classical open-label pharmacokinetic study of cisapride in neonates and young infants. METHODS: A total of 35 infants with a postconceptional age of 28 to 54 weeks at the time of the study received a single oral cisapride dose (0.2 mg/kg) at a postnatal age of 4 to 102 days, followed by repeated (n = 7) blood sampling over a 24-hour period. Cisapride and norcisapride were quantitated from plasma by HPLC-tandem mass spectrometry and pharmacokinetic data determined (n = 32) by noncompartmental methods. RESULTS: The pharmacokinetic parameters (mean +/- SD) were as follows: time to reach peak plasma concentration (t(max)), 4.4 +/- 2.8 hours (range, 0.9-12 hours); peak plasma concentration (C(max)), 29.3 +/- 16.6 ng/mL (range, 5.2-71.7 ng/mL); elimination half-life (t(1/2)), 10.7 +/- 3.7 hours (range, 1.9-18.1 hours); apparent total body clearance (Cl/F), 0.62 +/- 0.43 L. h(-1). kg(-1) (range, 0.2-1.9 L. h(-1). kg(-1)); and apparent volume of distribution (VD(ss)/F), 9.0 +/- 7.1 L/kg (range, 2.2-30.5 L/kg). The apparent renal clearance (CL(R)) of cisapride in infants (n = 28) was estimated to be 0.003 +/- 0.003 L. h(-1). kg(-1). Substratification of the population based on postconceptional age demonstrated the following findings for cisapride: (1) The mean (+/-SD) C(max) for cisapride was higher in the oldest postconceptional age category (44.5 +/- 19.6 ng/mL) than the middle and youngest categories (23.4 +/- 11.7 ng/mL and 30.0 +/- 17.5 ng/mL, respectively); (2) the t(max) for cisapride was shortest in the oldest postconceptional age category (2.2 +/- 1.1 hours) compared with the middle and youngest categories (4.4 +/- 3.3 hours and 5.0 +/- 2.6 hours, respectively); (3) the CL/F for cisapride in the youngest postconceptional age group was significantly lower (0.45 +/- 0.26 L. h(-1). kg(-1), P <.05) than in the middle and oldest categories (0.75 +/- 0.46 L. h(-1). kg(-1) and 0.85 +/- 0.69 L. h(-1). kg(-1), respectively); (4) a positive linear correlation was found between postconceptional age and the apparent terminal elimination rate constant (lambda(z)) for cisapride (P <.001, r(2) = 0.47) but not with CL/F. For norcisapride, the mean apparent C(max) was highest and the t(max) was shortest in the oldest postconceptional age group, although no association between postconceptional age and the norcisapride/cisapride area under the curve ratio was observed. All infants tolerated a single dose of cisapride well without significant alteration in QTc. CONCLUSIONS: (1) In neonates and infants, cisapride absorption and metabolism to its primary metabolite, norcisapride, were developmentally dependent; (2) approximately 99% of cisapride CL/F in neonates and young infants was nonrenal in nature; (3) CL/F of cisapride in neonates and infants noted in this study was reduced compared with data from older children and adults, likely as a result of developmental reductions in CYP3A4 activity; (4) as reflected by the correlation between postconceptional age and lambda(z), a rapid increase in total CYP3A4 activity occurs in the first 3 months of life.
Assuntos
Cisaprida/análogos & derivados , Cisaprida/farmacocinética , Sistema Enzimático do Citocromo P-450/metabolismo , Fármacos Gastrointestinais/farmacocinética , Área Sob a Curva , Cromatografia Líquida de Alta Pressão , Cisaprida/sangue , Cisaprida/metabolismo , Citocromo P-450 CYP3A , Feminino , Fármacos Gastrointestinais/sangue , Fármacos Gastrointestinais/metabolismo , Meia-Vida , Humanos , Lactente , Recém-Nascido , Masculino , Taxa de Depuração MetabólicaRESUMO
OBJECTIVES: Cisapride was compared with midazolam in vivo to determine its potential applicability as a cytochrome P450 (CYP) 3A4 "probe." As well, we evaluated whether cisapride was transported by P-glycoprotein. METHODS: Bidirectional transport assays were conducted in LLC-PK1 cells and the derivative cell line L-MDR1 to determine whether cisapride was a substrate for P-glycoprotein. A pharmacokinetic study was also conducted in 17 healthy adults (n = 8 women) who received intravenous midazolam (0.025 mg/kg), oral midazolam (0.15 mg/kg), and oral cisapride (0.07 mg/kg) in a randomized crossover design. Plasma concentrations were quantitated from repeated after-dosing blood samples by HPLC with ultraviolet detection for midazolam and HPLC with tandem mass spectrometry detection for cisapride and norcisapride. Pharmacokinetic parameters were determined by noncompartmental methods. Both linear and nonlinear regression analyses were used to examine the association between the apparent plasma clearance of midazolam and cisapride and the cisapride/norcisapride plasma concentration ratios. RESULTS: Although not a substrate for P-glycoprotein, cisapride inhibited P-glycoprotein with an apparent inhibition constant (K(i)) of 16.1 micromol/L. Linear correlations between cisapride clearance and both intravenous and oral midazolam clearance (P =.01, r(2) = 0.43 and P =.001, r(2) = 0.46, respectively) were found. Cisapride/norcisapride plasma concentration ratios at 8 hours (P =.001, r(2) = 0.90) and 12 hours (P =.001, r(2) = 0.96), as well as cisapride plasma concentrations at these time points, were shown to accurately predict the area under the plasma concentration versus time curve for cisapride. CONCLUSIONS: CYP3A4 activity reflected by the total body clearance after oral administration of cisapride should be independent of transport by P-glycoprotein. Concordance between the pharmacokinetics for cisapride and midazolam support the applicability of oral cisapride as a pharmacologic substrate to assess total CYP3A4 activity in vivo. Cisapride plasma concentration ratios at 8 or 12 hours after a single oral cisapride dose may prove useful as a single-point determination to reflect the area under the plasma concentration versus time curve and the plasma clearance of cisapride and, as well, total CYP3A4 activity in vivo.
Assuntos
Subfamília B de Transportador de Cassetes de Ligação de ATP/metabolismo , Cisaprida/análogos & derivados , Cisaprida/farmacocinética , Sistema Enzimático do Citocromo P-450/metabolismo , Subfamília B de Transportador de Cassetes de Ligação de ATP/antagonistas & inibidores , Administração Oral , Adolescente , Adulto , Animais , Área Sob a Curva , Cromatografia Líquida de Alta Pressão , Cisaprida/administração & dosagem , Cisaprida/sangue , Estudos Cross-Over , Citocromo P-450 CYP3A , Feminino , Humanos , Infusões Intravenosas , Células LLC-PK1/efeitos dos fármacos , Masculino , Espectrometria de Massas , Midazolam/administração & dosagem , Midazolam/sangue , Midazolam/farmacocinética , Pessoa de Meia-Idade , Modelos Biológicos , Análise de Regressão , SuínosRESUMO
Identification of the human cytochrome P450 (P450) enzymes involved in the metabolism of cisapride and racemic norcisapride [(+/-)-norcisapride] was investigated at 0.1 and 1 microM, concentrations that span the mean plasma C(max) for cisapride. Formation of norcisapride (Nor), 3-fluoro-4-hydroxycisapride (3F), and 4-fluoro-2-hydroxycisapride (4F) from cisapride and an uncharacterized metabolite (UNK) from (+/-)-norcisapride in human liver microsomes (HLMs) were consistent with Michaelis-Menten kinetics for a single enzyme (K(m), 6.0, 14.3, 13.9, and 107 microM; V(max), 1350, 696, 568, and 25 pmol/mg of protein, respectively). HLMs converted cisapride to Nor at rates that were at least 3 orders of magnitude greater than those observed for (+/-)-norcisapride conversion to UNK. The sample-to-sample variation in the rates of Nor, 3F, 4F, and UNK formation correlated strongly (r(2) > 0.796) with CYP3A4/5 activity in a panel of HLMs (n = 7) and was markedly reduced by ketoconazole, a potent CYP3A inhibitor. Ketoconazole virtually eliminated (+/-)-norcisapride conversion to UNK (94 +/- 0.5%). Studies with 10 cDNA-expressed enzymes revealed that CYP3A4 catalyzed the formation of Nor and 4F at rates >100 times those of non-CYP3A enzymes and >100- and 50-fold higher than CYP3A5 and CYP3A7, respectively. CYP3A4 was the only P450 capable of UNK formation. Therefore, CYP3A4 is the principal P450 enzyme responsible for the conversion of cisapride to Nor, 3F, and 4F and of (+/-)-norcisapride to UNK. Compared with cisapride, factors related to CYP3A4-mediated (+/-)-norcisapride metabolism (e.g., ontogeny of drug-metabolizing enzymes, inhibition, and induction) should be clinically unimportant due to the apparent lack of dependence on cytochromes P450 for elimination.
Assuntos
Antiulcerosos/farmacocinética , Hidrocarboneto de Aril Hidroxilases , Cisaprida/análogos & derivados , Cisaprida/farmacocinética , Sistema Enzimático do Citocromo P-450/metabolismo , Oxirredutases N-Desmetilantes/metabolismo , Biotransformação , Cromatografia Líquida de Alta Pressão , Citocromo P-450 CYP3A , Humanos , Técnicas In Vitro , Isoenzimas/metabolismo , Masculino , Espectrometria de Massas , EstereoisomerismoRESUMO
OBJECTIVES: Our objective was to compare the interactions of red wine and grapefruit juice with cisapride. METHODS: The oral pharmacokinetics of cisapride, its norcisapride metabolite, and electrocardiographic QTc interval were determined over a 24-hour period after administration of cisapride 10 mg with 250 mL grapefruit juice, red wine (cabernet sauvignon), or water in a randomized 3-way crossover study in 12 healthy men. RESULTS: The cisapride area under the concentration-time curve (AUC) and the maximum plasma drug concentration after single-dose administration (C(max)) with grapefruit juice were 151% (P <.01) and 168% (P <.001), respectively, of those with water. The increase in cisapride AUC and C(max) was variable among individuals; however, cisapride AUC and C(max) were enhanced by the same proportion. The time to reach maximum concentration after drug administration (t(max)) and the apparent elimination half-life (t((1/2)) for cisapride and the pharmacokinetics of norcisapride were not altered. Norcisapride/cisapride ratios were reduced. Cisapride AUC and C(max) with red wine were 115% (difference not statistically significant) and 107% (difference not statistically significant), respectively, of those with water. The cisapride t(max) was slightly longer. Cisapride t((1/2)) and norcisapride pharmacokinetics were not different. The norcisapride/cisapride ratio at cisapride C(max) was lower. One subject had a doubling in cisapride AUC and C(max) and a decrease in norcisapride/cisapride ratios with red wine and also had the largest interaction with grapefruit juice. QTc interval was unchanged in all treatment groups and individuals. CONCLUSIONS: A single glass of grapefruit juice produced an individual-dependent variable increase in the systemic availability of cisapride by inhibition of intestinal cytochrome P450 3A4 (CYP3A4) activity. The identical volume of red wine caused only minor changes in cisapride pharmacokinetics despite some inhibition of CYP3A4 in most individuals. However, even this amount of red wine may cause a marked interaction similar to that for grapefruit juice in individuals with a preexisting high intestinal CYP3A4 content.
Assuntos
Bebidas , Cisaprida/análogos & derivados , Cisaprida/farmacocinética , Citrus , Inibidores das Enzimas do Citocromo P-450 , Fármacos Gastrointestinais/farmacocinética , Oxigenases de Função Mista/antagonistas & inibidores , Vinho , Adulto , Área Sob a Curva , Cisaprida/sangue , Cisaprida/farmacologia , Estudos Cross-Over , Citocromo P-450 CYP3A , Interações Alimento-Droga , Fármacos Gastrointestinais/sangue , Fármacos Gastrointestinais/farmacologia , Humanos , Masculino , Valores de ReferênciaRESUMO
A simple high-performance liquid chromatography assay using fluorescence detection for the major metabolite of the gastric prokinetic drug cisapride, norcisapride, is presented. Analysis is performed using an Alltech Platinum EPS C8 column with a mobile phase made up of methanol and 0.02M sodium dihygrogen phosphate (45:55, v/v) containing triethylamine (1 g/L). Complete resolution is achieved among norcisapride, the internal standard (metoclopramide), and endogenous urinary components. The assay is linear over the range 50-2000 ng/mL with a mean recovery of 71.2% across the analytical range following solvent extraction with toluene-isoamyl alcohol (95:5, v/v). Intraday coefficients of variation (precision) determined at 200 and 1000 ng/mL are 6.0 and 9.8%, respectively, and interday coefficients of variation are 8.8 and 6.6%, respectively. Intra- and interassay accuracy (as mean relative error) determined at the same concentrations is within 10% in all cases. An analysis of urine samples from a healthy volunteer following the administration of a single 10-mg oral dose of cisapride is shown.
Assuntos
Cisaprida/análogos & derivados , Cromatografia Líquida de Alta Pressão , Cisaprida/urina , Humanos , Indicadores e Reagentes , Reprodutibilidade dos TestesRESUMO
A simple, rapid and reproducible high-performance liquid chromatographic assay for cisapride and norcisapride in human plasma is described. Samples of plasma (150 microl) were extracted using a C18 solid-phase cartridge. Regenerated tubes were eluted with 1.0 ml of methanol, dried, redissolved in 150 microl of methanol and injected. Chromatography was performed at room temperature by pumping acetonitrile-methanol-0.015 M phosphate buffer pH 2.2-2.3 (680:194:126, v/v/v) at 0.8 ml/min through a C18 reversed-phase column. Cisapride, norcisapride and internal standard were detected by absorbance at 276 nm and were eluted at 4.3, 5.3 and 8.1 min, respectively. Calibration plots in plasma were linear (r>0.998) from 10 to 150 ng/ml. Intraday precisions for cisapride and norcisapride were 3.3% and 5.4%, respectively. Interday precisions for cisapride and norcisapride were 9.6% and 9.0%, respectively. Drugs used which might be coadministered were tested for interference.