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Antivirales , Tratamiento Farmacológico de COVID-19 , Ritonavir , SARS-CoV-2 , Humanos , Ritonavir/uso terapéutico , Ritonavir/administración & dosificación , Antivirales/uso terapéutico , Masculino , Persona de Mediana Edad , Femenino , COVID-19 , Tiempo de Tratamiento , Anciano , Leucina/uso terapéutico , Resultado del Tratamiento , Retraso del Tratamiento , Lactamas , Nitrilos , ProlinaRESUMEN
CONTEXT: Peroxisome proliferator-activated receptor-γ (PPARγ) is a member of the nuclear hormone receptor superfamily of ligand-activated transcription factors. Activation of PPARγ pathway has been shown to enhance fatty acid oxidation, improve endothelial cell function, and decrease myocardial fibrosis in heart failure. Thus, the protein has been raised as an attractive target for heart failure therapy. OBJECTIVE: This work attempted to discover new and potent PPARγ agonists from natural products using a synthetic strategy of computer virtual screening and transactivation reporter assay. MATERIALS AND METHODS: A large library of structurally diverse, drug-like natural products was compiled, from which those with unsatisfactory pharmacokinetic profile and/or structurally redundant compounds were excluded. The binding mode of remaining candidates to PPARγ ligand-binding domain (LBD) was computationally modelled using molecular docking and their relative binding potency was ranked by an empirical scoring scheme. Consequently, eight commercially available hits with top scores were selected and their biological activity was determined using a cell-based reporter-gene assay. RESULTS: Four natural product compounds, namely ZINC13408172, ZINC4292805, ZINC44179 and ZINC901461, were identified to have high or moderate agonistic potency against human PPARγ with EC50 values of 0.084, 2.1, 0.35 and 5.6 µM, respectively, which are comparable to or even better than that of the approved PPARγ full agonists pioglitazone (EC50 = 0.16 µM) and rosiglitazone (EC50 = 0.034 µM). Hydrophobic interactions and van der Waals contacts are the primary chemical forces to stabilize the complex architecture of PPARγ LBD domain with these agonist ligands, while few hydrogen bonds, salt bridges and/or π-π stacking at the complex interfaces confer selectivity and specificity for the domain-agonist recognition. DISCUSSION AND CONCLUSION: The integrated in vitro-in silico screening strategy can be successfully applied to rational discovery of biologically active compounds. The newly identified natural products with PPARγ agonistic potency are considered as promising lead scaffolds to develop novel chemical therapeutics for heart failure.
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Productos Biológicos/farmacología , Fármacos Cardiovasculares/farmacología , Descubrimiento de Drogas/métodos , Insuficiencia Cardíaca/tratamiento farmacológico , Ensayos Analíticos de Alto Rendimiento , PPAR gamma/agonistas , Animales , Sitios de Unión , Productos Biológicos/química , Productos Biológicos/metabolismo , Productos Biológicos/farmacocinética , Células CACO-2 , Fármacos Cardiovasculares/química , Fármacos Cardiovasculares/metabolismo , Fármacos Cardiovasculares/farmacocinética , Perros , Genes Reporteros , Insuficiencia Cardíaca/metabolismo , Insuficiencia Cardíaca/fisiopatología , Humanos , Enlace de Hidrógeno , Ligandos , Células de Riñón Canino Madin Darby , Simulación del Acoplamiento Molecular , Estructura Molecular , PPAR gamma/genética , PPAR gamma/metabolismo , Unión Proteica , Reproducibilidad de los Resultados , Transducción de Señal/efectos de los fármacos , Relación Estructura-Actividad , TransfecciónRESUMEN
OBJECTIVE: To investigate whether rs12731181 (AâG) interrupted miR-590-3p-mediated suppression of the prostaglandin F2α receptor (FP) and whether it is associated with essential hypertension in the Chinese population. APPROACH AND RESULTS: We found that miR-590-3p regulates human FP gene expression by binding to its 3'-untranslated region. rs12731181 (AâG) altered the binding affinity between miR-590-3p and its FP 3'-untranslated region target, thus reducing the suppression of FP expression, which, in turn, enhanced FP receptor-mediated contractility of vascular smooth muscle cells. Overexpression of FP augmented vascular tone and elevated blood pressure in mice. An association study was performed to analyze the relationship between the FP gene and essential hypertension in the Han Chinese population. The results indicated that the rs12731181 G allele was associated with susceptibility to essential hypertension. Carriers of the AG genotype exhibited significantly higher blood pressure than those of the AA genotype. FP gene expression was significantly higher in human peripheral leukocytes from individuals with the AG genotype than that in leukocytes from individuals with the AA genotype. CONCLUSIONS: rs12731181 in the seed region of the miR-590-3p target site is associated with increased risk of essential hypertension and represents a new paradigm for FP involvement in blood pressure regulation.
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Pueblo Asiatico/genética , Hipertensión/genética , MicroARNs/genética , Receptores de Prostaglandina/genética , Regiones no Traducidas 3' , Animales , Sitios de Unión , China/etnología , Hipertensión Esencial , Predisposición Genética a la Enfermedad , Humanos , Ratones , Polimorfismo de Nucleótido Simple , Transcripción GenéticaRESUMEN
OBJECTIVE: Lipoprotein-associated phospholipase A2 (Lp-PLA2) is a useful inflammatory marker of cardiovascular risk, yet there are few reports on the prognostic significance of Lp-PLA2 as a risk factor for acute coronary syndrome (ACS). It is necessary to evaluate the association of Lp-PLA2 with major adverse cardiac events (MACE) in patients with ACS and assess its incremental value for risk discrimination over established risk factors and biomarkers. METHODS: One hundred fifty-two patients with ACS and 142 patients without coronary artery disease (CAD) from Shanghai Xuhui District, PRC (aged < 80 years) were enrolled from February 2007 to March 2008 and were followed for a median of 6 months (range, 4-10 months). Plasma Lp-PLA2 activity was measured at baseline with liquid chromatography tandem mass spectrometry. Its clinical significance was evaluated with existing risk indicators. RESULTS: Lp-PLA2 activity was higher in patients with ACS than in patients without CAD (22.36 +/- 1.23 mg/mL vs 19.74 +/- 3.85 mg/mL; P = 0.027). During the follow-up period, 5 cases of cardiovascular death, 8 cases of nonfatal myocardial infarction, and 11 cases of target vessel revascularization occurred. Elevated Lp-PLA2 was associated with an increased risk of MACE (hazard ratio, 1.52; 95% confidence interval, 1.09-2.37; P = 0.033). The Lp-PLA2 activity level in incidental cases was higher than in nonincidental cases (P = 0.04). CONCLUSION: In this community-based cohort of patients with ACS, Lp-PLA2 was strongly and independently associated with major adverse cardiac events and contributed incrementally to risk discrimination.
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1-Alquil-2-acetilglicerofosfocolina Esterasa/sangre , Síndrome Coronario Agudo/enzimología , Anciano , Biomarcadores/sangre , Cromatografía Liquida , Estudios de Cohortes , Femenino , Humanos , Masculino , Persona de Mediana Edad , Fosfolipasas A2 , Pronóstico , Factores de Riesgo , Espectrometría de Masas en TándemRESUMEN
OBJECTIVE: Here we develop a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the determination of digoxin in serum. DESIGN AND METHODS: The serum samples were extracted with methyl tert-butyl ether using an isotope-labeled digoxin-d3 as internal standard. The analyte was separated on a reverse phase Capcell C18 column and detected in positive electrospray ionization multiple reaction monitoring mass spectrometry. RESULTS: The chromatographic analysis was carried out within 3 min, but the complete analysis took longer because of the liquid-liquid extraction. The lower limit of quantification was 0.1 ng/mL for digoxin. The intra- and inter-batch precisions were less than 12%, and the bias ranged from -9.1% to 10.7%. The external quality assessment (EQA) results obtained with the LC-MS/MS method were comparable to target values. Subsequently, this method has been applied to the therapeutic monitoring of digoxin in a clinical setting. CONCLUSION: In this study, we have developed a rapid and reliable LC-MS/MS method for the therapeutic monitoring of digoxin in human serum.
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Cromatografía Liquida/métodos , Digoxina/sangre , Insuficiencia Cardíaca/sangre , Espectrometría de Masas en Tándem/métodos , Digoxina/uso terapéutico , Insuficiencia Cardíaca/tratamiento farmacológico , Humanos , Estructura Molecular , Control de Calidad , Reproducibilidad de los ResultadosRESUMEN
OBJECTIVE: To investigate the effects of pravastatin, fosinopril and their combination on ventricular remodeling, cardiac function, tumor necrosis factor-alpha (TNF-alpha) mRNA expression, and matrix metalloproteinases (MMPs) activities after myocardial infarction (MI) in rats. METHODS: Acute myocardial infarction (AMI) was established by ligation of the anterior descending coronary artery in male Sprague-Dawly (SD) rats. Twenty-four hours after the procedure, the 48 surviving rats were grouped randomly as AMI control, fosinopril (10 mg.kg(-1).d(-1)), pravastatin (20 mg.kg(-1).d(-1)) and a combined use of the 2 drugs. Sham-operated group (n = 8) was taken randomly as non-infarction control. Six weeks after treatment with the drugs by gastric gavage, heart function and left ventricular remodeling were assessed. Left ventricular weight (LVW)/body weight (BW) ratio was determined. The relative expression of myocardium TNF-alpha mRNA was assessed by reverse transcription-polymerase chain reaction. Left ventricular myocardium MMPs activities were assessed by Zymography. RESULTS: There were no significant differences among the four AMI groups in infarction size (P > 0.05). In comparison with the AMI group, left ventricular end-diastolic pressure, left ventricular end-diastolic diameter, LVW/BW all decreased significantly (P < 0.05 - 0.01); while dp/dtmax, dp/dtmin, fractional shortening (FS) and ejection fraction (EF) increased significantly in all three drug-treated groups (P < 0.05 - 0.01); increments of FS, LVEF and dp/dtmax were more evident in the combination group than either the fosinopril or pravastatin group (P < 0.05). The levels of TNF-alpha mRNA in AMI rats treated with fosinopril, pravastatin and their combination reduced 29%, 26% and 33%, respectively (P < 0.01); MMP-2 activity reduced 25%, 30% and 35%, respectively (P < 0.01); MMP-9 activity reduced 20%, 18% and 24%, respectively (P < 0.01). There were no significant differences in other variables among the 3 treatment groups (P > 0.05). CONCLUSION: Pravastatin, fosinopril and their combination showed favorable effects on left ventricular remodeling after AMI in rats and demonstrated improved cardiac function. The combined treatment group yielded better results in the context of improving left ventricular systolic function. These effects could be relevant to the attenuation of increased MMP-2 and MMP-9 activities and left ventricular expression of TNF-alpha.