RESUMO
BACKGROUND: CYP2D6*10 is mainly responsible for the large pharmacokinetic variability of routinely administered metoprolol in middle-aged and elderly Asian patients. Utilizing an efficient method for identifying the CYP2D6*10 genotypes is clinically important for evaluating the pharmacokinetic effect of administration of metoprolol. This study attempted to evaluate the effectiveness of the two methods used to detect the rs1065852 and rs1135840 SNPs of the CYP2D6*10 gene. METHODS: Blood samples were processed for the collection of genomic DNA from 198 subjects across Chinese population, and detection of CYP2D6*10 (rs1065852 and rs1135840) was performed using the PyroMark Q24 pyrose-quencing and matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry (MALDI-TOF MS). The discordant results were further validated with Sanger sequencing. We eventually attempted to assess some features of these two methods including reliability, rapidness, being appropriate, and cost-effectiveness. RESULTS: Genotyping of rs1065852 and rs1135840 detected by MALDI-TOF MS were concordant with those identified by PyroMark Q24 pyrosequencing in all 198 (100%) individuals. The hands-on-time and the turnaround time were shorter in the PyroMark Q24 pyrosequencing method than in the MALDI-TOF MS method for SNP of CYP2D6*10. In terms of being cost-effective and high-throughput, the MALDI-TOF MS method outperformed the PyroMark Q24 pyrosequencing method. CONCLUSIONS: CYP2D6*10 genotypes detected by PyroMark Q24 pyrosequencing and MALDI-TOF-MS showed that both methods were reliable, rapid, appropriate, and cost-effective methods. These methods are valuable for clinical applications.
Assuntos
Citocromo P-450 CYP2D6/genética , Técnicas de Genotipagem/métodos , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Polimorfismo de Nucleotídeo Único , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Idoso , Anti-Hipertensivos/sangue , Anti-Hipertensivos/metabolismo , Anti-Hipertensivos/uso terapêutico , Povo Asiático/genética , China , Análise Custo-Benefício , Citocromo P-450 CYP2D6/metabolismo , Feminino , Genótipo , Técnicas de Genotipagem/economia , Humanos , Hipertensão/sangue , Hipertensão/tratamento farmacológico , Hipertensão/genética , Masculino , Metoprolol/sangue , Metoprolol/metabolismo , Metoprolol/uso terapêutico , Pessoa de Meia-Idade , Reprodutibilidade dos TestesRESUMO
Hypoxic pulmonary vasoconstriction (HPV) is a well-characterized vascular response to low oxygen pressures and is involved in life-threatening conditions such as high-altitude pulmonary edema (HAPE) and pulmonary arterial hypertension (PAH). While the efficacy of oral therapies can be affected by drug metabolism, or dose-limiting systemic toxicity, inhaled treatment via pressured metered dose inhalers (pMDI) may be an effective, nontoxic, practical alternative. We hypothesized that a stable water-in-perfluorooctyl bromide (PFOB) emulsion that provides solubility in common pMDI propellants, engineered for intrapulmonary delivery of pulmonary vasodilators, reverses HPV during acute hypoxia (HX). Male Sprague Dawley rats received two 10-min bouts of HX (13% O2) with 20 min of room air and drug application between exposures. Treatment groups: intrapulmonary delivery (PUL) of (1) saline; (2) ambrisentan in saline (0.1 mg/kg); (3) empty emulsion; (4) emulsion encapsulating ambrisentan or sodium nitrite (NaNO2) (0.1 and 0.5 mg/kg each); and intravenous (5) ambrisentan (0.1 mg/kg) or (6) NaNO2 (0.5 mg/kg). Neither PUL of saline or empty emulsion, nor infusions of drugs prevented pulmonary artery pressure (PAP) elevation (32.6 ± 3.2, 31.5 ± 1.2, 29.3 ± 1.8, and 30.2 ± 2.5 mmHg, respectively). In contrast, PUL of aqueous ambrisentan and both drug emulsions reduced PAP by 20-30% during HX, compared to controls. IL6 expression in bronchoalveolar lavage fluid and whole lung 24 h post-PUL did not differ among cohorts. We demonstrate proof-of-concept for delivering pulmonary vasodilators via aerosolized water-in-PFOB emulsion. This concept opens a potentially feasible and effective route of treating pulmonary vascular pathologies via pMDI.
Assuntos
Sistemas de Liberação de Medicamentos/métodos , Emulsões/administração & dosagem , Fluorocarbonos/administração & dosagem , Hipertensão Pulmonar/tratamento farmacológico , Edema Pulmonar/tratamento farmacológico , Água/administração & dosagem , Animais , Anti-Hipertensivos/administração & dosagem , Anti-Hipertensivos/metabolismo , Avaliação Pré-Clínica de Medicamentos/métodos , Emulsões/metabolismo , Fluorocarbonos/metabolismo , Hipertensão Pulmonar/diagnóstico por imagem , Hipertensão Pulmonar/metabolismo , Masculino , Fenilpropionatos/administração & dosagem , Fenilpropionatos/metabolismo , Circulação Pulmonar/efeitos dos fármacos , Circulação Pulmonar/fisiologia , Edema Pulmonar/diagnóstico por imagem , Edema Pulmonar/metabolismo , Piridazinas/administração & dosagem , Piridazinas/metabolismo , Ratos , Ratos Sprague-Dawley , Resultado do Tratamento , Água/metabolismoRESUMO
Bosentan is a dual endothelin receptor antagonist used in the treatment of pulmonary arterial hypertension (PAH). But the solubility and bioavailability of this drug are poor, which has restricted the design and development of dosage forms for efficient and successful therapy. The present study was carried out to develop nanocomposites using an amphiphilic graft co-polymer (Soluplus®) as a carrier to enhance the solubility and bioavailability of bosentan. The graft co-polymer-based nanocomposite formulation was prepared using the single-emulsion technique. The nanocomposite was characterised in terms of particle size analysis, solubility, percentage entrapment efficiency, drug-loading capacity, surface morphology, drug content, in vitro dissolution, stability and bioavailability. FT-IR study revealed that there was no interaction between the drug and Soluplus®. DSC analysis of the nanocomposite formulation confirmed that the bosentan was completely encapsulated within a Soluplus®. XRD analysis showed that the drug was converted to an amorphous form irreversibly. SEM images showed that the particles were of size 96-129µm and had slightly smooth to rough textured surface. TEM analysis indicated that the diameters of the prepared bosentan nanocomposite after dispersion in distilled water were 13.69-96.78nm. Statistically significant increases in the solubility, dissolution and bioavailability of the drug were observed. It was confirmed that the use of a graft co-polymer carrier-based nanocomposite formulation is a good approach for efficient delivery of bosentan, the solubility and bioavailability being increased manifold.
Assuntos
Portadores de Fármacos/química , Nanocompostos/química , Polímeros/química , Sulfonamidas/química , Tensoativos/química , Animais , Anti-Hipertensivos/metabolismo , Disponibilidade Biológica , Bosentana , Portadores de Fármacos/metabolismo , Estabilidade de Medicamentos , Feminino , Masculino , Polímeros/metabolismo , Coelhos , Solubilidade , Sulfonamidas/metabolismo , Tensoativos/metabolismo , Difração de Raios XRESUMO
In this article, we consider the new drugs approved for the European market in 2015. We present a summary of the new mechanisms of action introduced and highlight three new mechanisms of action with a potentially high future impact: PCSK9 inhibition (alirocumab (Praluent®) and evolocumab (Repatha®)) for hypercholesterolaemia, neprilysin inhibition (sacubitril in combination with valsartan (Entresto®)) for heart failure, and interleukin-5 inhibition (mepolizumab (Nucala®)) for asthma.
Assuntos
Antiasmáticos , Anticorpos Monoclonais Humanizados , Anticorpos Monoclonais , Anti-Hipertensivos , Hipolipemiantes , Antiasmáticos/metabolismo , Antiasmáticos/farmacologia , Anticorpos Monoclonais/metabolismo , Anticorpos Monoclonais/farmacologia , Anticorpos Monoclonais Humanizados/metabolismo , Anticorpos Monoclonais Humanizados/farmacologia , Anti-Hipertensivos/metabolismo , Anti-Hipertensivos/farmacologia , Aprovação de Drogas , Indústria Farmacêutica , Europa (Continente) , Humanos , Hipolipemiantes/metabolismo , Hipolipemiantes/farmacologia , Transdução de Sinais/efeitos dos fármacosRESUMO
Bioactive peptides are food derived components, usually consisting of 3-20 amino acids, which are inactive when incorporated within their parent protein. Once liberated by enzymatic or chemical hydrolysis, during food processing and gastrointestinal transit, they can potentially provide an array of health benefits to the human body. Owing to an unprecedented increase in the worldwide incidence of obesity and hypertension, medical researchers are focusing on the hypotensive and anti-obesity properties of nutritionally derived bioactive peptides. The role of the renin-angiotensin system has long been established in the aetiology of metabolic diseases and hypertension. Targeting the renin-angiotensin system by inhibiting the activity of angiotensin-converting enzyme (ACE) and preventing the formation of angiotensin II can be a potential therapeutic approach to the treatment of hypertension and obesity. Fish-derived proteins and peptides can potentially be excellent sources of bioactive components, mainly as a source of ACE inhibitors. However, increased use of marine sources, poses an unsustainable burden on particular fish stocks, so, the underutilized fish species and by-products can be exploited for this purpose. This paper provides an overview of the techniques involved in the production, isolation, purification, and characterization of bioactive peptides from marine sources, as well as the evaluation of the ACE inhibitory (ACE-I) activity and bioavailability.
Assuntos
Fármacos Antiobesidade/uso terapêutico , Anti-Hipertensivos/uso terapêutico , Organismos Aquáticos/química , Descoberta de Drogas , Fragmentos de Peptídeos/uso terapêutico , Animais , Fármacos Antiobesidade/economia , Fármacos Antiobesidade/isolamento & purificação , Fármacos Antiobesidade/metabolismo , Anti-Hipertensivos/economia , Anti-Hipertensivos/isolamento & purificação , Anti-Hipertensivos/metabolismo , Proteínas Alimentares/química , Proteínas Alimentares/isolamento & purificação , Proteínas Alimentares/metabolismo , Proteínas Alimentares/uso terapêutico , Suplementos Nutricionais/economia , Descoberta de Drogas/tendências , Proteínas de Peixes/química , Proteínas de Peixes/isolamento & purificação , Proteínas de Peixes/metabolismo , Proteínas de Peixes/uso terapêutico , Indústria de Processamento de Alimentos/economia , Humanos , Hipertensão/dietoterapia , Hipertensão/tratamento farmacológico , Resíduos Industriais/análise , Resíduos Industriais/economia , Obesidade/dietoterapia , Obesidade/tratamento farmacológico , Oligopeptídeos/economia , Oligopeptídeos/isolamento & purificação , Oligopeptídeos/metabolismo , Oligopeptídeos/uso terapêutico , Fragmentos de Peptídeos/economia , Fragmentos de Peptídeos/isolamento & purificação , Fragmentos de Peptídeos/metabolismo , ProteóliseRESUMO
BACKGROUND: Bee larvae are considered to be an important reservoir for proteins. However, little attention has been paid to the release of potential bioactive peptides from bee larva proteins. In this study the major protein in bee larvae was hydrolyzed in vitro by gastrointestinal enzymes. The peptide profile of the hydrolysis was characterized by gel filtration chromatography and tricine-SDS-PAGE. Furthermore, the bioactive peptide was isolated and identified by Q-TOF-MS/MS. RESULTS: The major bee larva protein was identified as apalbumin 2 and was more digestible into peptides with molecular weights lower than 3 kDa. The hydrolysate obtained after 3 h of digestion exhibited angiotensin I-converting enzyme (ACE)-inhibitory activity and was purified sequentially by gel filtration and RP-HPLC. The molecular weights of peptide fractions with ACE-inhibitory activity were distributed between 0.5 and 1.5 kDa. A novel peptide with highest ACE-inhibitory activity (IC50 54.9 µmol L(-1) ) was purified by further RP-HPLC. The amino acid sequence of this peptide was identified as LLKPY (632.40 Da). CONCLUSION: ACE-inhibitory peptides could be formed from bee larvae through gastrointestinal digestion. The most active peptide (LLKPY) is potentially useful as a therapeutic agent in treating hypertension.
Assuntos
Inibidores da Enzima Conversora de Angiotensina/isolamento & purificação , Digestão , Descoberta de Drogas , Proteínas de Insetos/isolamento & purificação , Modelos Biológicos , Oligopeptídeos/isolamento & purificação , Fragmentos de Peptídeos/isolamento & purificação , Sequência de Aminoácidos , Inibidores da Enzima Conversora de Angiotensina/química , Inibidores da Enzima Conversora de Angiotensina/metabolismo , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Animais , Anti-Hipertensivos/química , Anti-Hipertensivos/isolamento & purificação , Anti-Hipertensivos/metabolismo , Anti-Hipertensivos/farmacologia , Criação de Abelhas/economia , Abelhas/química , China , Quimotripsina/metabolismo , Proteínas de Insetos/química , Proteínas de Insetos/economia , Proteínas de Insetos/metabolismo , Proteínas de Insetos/farmacologia , Larva/química , Peso Molecular , Oligopeptídeos/química , Oligopeptídeos/metabolismo , Oligopeptídeos/farmacologia , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/metabolismo , Fragmentos de Peptídeos/farmacologia , Hidrolisados de Proteína/química , Hidrolisados de Proteína/economia , Hidrolisados de Proteína/metabolismo , Hidrolisados de Proteína/farmacologia , Proteólise , Tripsina/metabolismo , Resíduos/análise , Resíduos/economiaRESUMO
Since many years, numerous kinds of processes based on enzymatic hydrolysis at various pH, involving added plant or bacterial enzymes after inactivation by heating of endogenous enzymes present in the raw material or, alternatively, based on the action of endogenous enzymes, have contributed to the degradation of marine by-product proteins in order to produce fractions exerting biological activities. Peptides obtained by enzymatic hydrolysis of fish proteins exhibit not only nutritional but also biological properties of dietary uses, or even therapeutic potential. In this review, we have focused on the different enzymatic processes able to generate bioactive peptides from marine by-products and exerting high potential in nutraceutical applications to fight against important public health issues like obesity, stress, hypertension, and migraine. Beyond the nutraceutical and pharmaceutical aspects, this way of valorization is also included in the necessary development of by-product fishing industries for economic and ecological reasons in the worldwide context of marine resources depletion.
Assuntos
Suplementos Nutricionais , Peixes/metabolismo , Promoção da Saúde , Resíduos Industriais/análise , Animais , Anti-Hipertensivos/economia , Anti-Hipertensivos/metabolismo , Anti-Hipertensivos/farmacologia , Anti-Hipertensivos/uso terapêutico , Antioxidantes/economia , Antioxidantes/metabolismo , Antioxidantes/farmacologia , Antioxidantes/uso terapêutico , Colagogos e Coleréticos/economia , Colagogos e Coleréticos/metabolismo , Colagogos e Coleréticos/farmacologia , Colagogos e Coleréticos/uso terapêutico , Suplementos Nutricionais/economia , Descoberta de Drogas , Proteínas de Peixes/metabolismo , Pesqueiros/economia , Humanos , Resíduos Industriais/economia , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/metabolismo , Fragmentos de Peptídeos/farmacologia , Fragmentos de Peptídeos/uso terapêutico , Hidrolisados de Proteína/química , Hidrolisados de Proteína/metabolismo , Hidrolisados de Proteína/farmacologia , Hidrolisados de Proteína/uso terapêuticoRESUMO
PURPOSE: The objective of this study was to investigate the absorption behavior of total flavones of Hippophae rhamnoides L. (TFH) (the sum of isorhamnetin and quercetin as the index component) in the rat intestine using in situ circulation method. METHODS: The accumulated TFH absorption and related absorption parameters were calculated. Furthermore, the influences of Cremophor ELP and the P-glycoprotein inhibitor, verapamil, on the intestinal absorption of TFH were studied using the in situ circulation model. RESULTS AND DISCUSSION: The results showed that the absorption of TFH increased linearly with its concentration, indicating that a passive diffusion process was dominated. There were no significant differences in the absorption of TFH in three small intestine segments of duodenum, jejunum, and ileum and at different concentrations of Cremophor ELP ranging from 0.25% to 1% (P > 0.05). With the presence of P-gp inhibitor, verapamil, in the circulation fluid, the accumulated absorption of TFH did not increase significantly (P > 0.05). Further studies on the solubility and permeability enhancement of TFH should be investigated to develop new TFH products with high bioavailability.
Assuntos
Anti-Hipertensivos/metabolismo , Flavonas/farmacocinética , Hippophae/química , Hippophae/metabolismo , Absorção Intestinal , Intestino Delgado/metabolismo , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/antagonistas & inibidores , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/química , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Absorção , Animais , Anti-Hipertensivos/química , Anti-Hipertensivos/farmacocinética , Anti-Hipertensivos/farmacologia , Disponibilidade Biológica , Estabilidade de Medicamentos , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/farmacocinética , Medicamentos de Ervas Chinesas/farmacologia , Flavonas/química , Flavonas/farmacologia , Absorção Intestinal/efeitos dos fármacos , Masculino , Polietilenoglicóis/química , Polietilenoglicóis/metabolismo , Ratos , Ratos Sprague-Dawley , Verapamil/farmacologiaRESUMO
Nicotianamine (NA), a metal chelator, is ubiquitous in higher plants. In humans, NA inhibits angiotensin I-converting enzyme (ACE), and consequently reduces high blood pressure. Nicotianamine is synthesized from the trimerization of S-adenosylmethionine (SAM) by NA synthase (NAS). Here, we aimed to produce large amounts of NA fermentatively by introducing the Arabidopsis AtNAS2 gene into Saccharomyces cerevisiae strain SCY4. This strain can accumulate up to 100 times the usual amount of SAM, and this is considered desirable for overproduction of NA. Nicotianamine was produced in the engineered yeast, and the NA level increased with incubation time until the stationary phase. The maximum concentration of intracellular NA obtained was 766+/-33 microg/g wet weight. Successful production of NA in S. cerevisiae should pave the way for industrial production of this novel antihypertensive substrate.
Assuntos
Anti-Hipertensivos/metabolismo , Ácido Azetidinocarboxílico/análogos & derivados , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Alquil e Aril Transferases/genética , Alquil e Aril Transferases/metabolismo , Anti-Hipertensivos/química , Ácido Azetidinocarboxílico/química , Ácido Azetidinocarboxílico/metabolismo , Proliferação de Células , Meios de Cultura , Indústria Farmacêutica , Dosagem de Genes/genética , Regulação Fúngica da Expressão Gênica , Vetores Genéticos/genética , Estrutura Molecular , Engenharia de Proteínas , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Saccharomyces cerevisiae/química , Saccharomyces cerevisiae/citologiaRESUMO
The pharmacokinetic profiles of two brands of losartan 50 mg tablets were compared in 24 healthy adult volunteers after a single oral dose in a randomized cross-over study. The study was conducted at the ACDIMA Center for Bioequivalence & Pharmaceutical Studies, Amman, Jordan. The reference (Cozaar, MSD, The Netherlands) and test (Blosart, Julphar, UAE) products were administered to fasting volunteers. Blood samples were collected at specified time intervals, and the plasma separated and analysed for losartan and its active metabolite (losartan carboxylic acid) using a validated HPLC method with fluorescence detection. Pharmacokinetic parameters AUC(0-t), AUC(0-alpha), C(max), T(max), T(1/2), elimination rate constant, MRT, Cl/F and Vss/F were determined from plasma concentration-time profiles of both formulations and found to be in good agreement with reported values. Three parameters (AUC(0-t), AUC(0-alpha), and C(max)) were compared statistically to evaluate the bioequivalence between the two brands, using statistical modules recommended by the FDA. Analysis of variance (ANOVA) did not show any significant difference between the two formulations and 90% confidence intervals fell within the acceptable range (80%-125%) for bioequivalence. Based on these statistical inferences it was concluded that the two formulations exhibited comparable pharmacokinetic profiles and that Julphar's Blosart is bioequivalent to Cozaar of MSD, The Netherlands.
Assuntos
Anti-Hipertensivos/farmacocinética , Losartan/farmacocinética , Adulto , Análise de Variância , Anti-Hipertensivos/sangue , Anti-Hipertensivos/metabolismo , Área Sob a Curva , Química Farmacêutica , Estudos Cross-Over , Meia-Vida , Humanos , Jordânia , Losartan/sangue , Losartan/metabolismo , Masculino , Comprimidos , Equivalência TerapêuticaRESUMO
Long-term adherence or compliance with anti-hypertensive drug therapy is poor. It has been estimated that within the first year of treatment 16-50% of hypertensives discontinue their anti-hypertensive medications. Even among those who remain on therapy long term, missed medication doses are common. Epidemiological studies have shown that drug-treated hypertensives have higher blood pressures than age-, gender- and body mass index-matched normotensives. In addition, drug-treated hypertensive men and women who achieve blood pressure normalization are less likely to die over a 9.5-year period than those whose blood pressure remains elevated while taking anti-hypertensive drugs. Thus, one reason for less than optimal reduction of blood pressure-related cardiovascular-renal risk in drug-treated hypertensives is inadequate blood pressure lowering. Quantifiable excess risk has been documented even in the short term ( < 1 year) after interruption or discontinuation of anti-hypertensive medications as total healthcare costs are higher, mostly because of higher hospitalization rates. Data from the Treatment of Mild Hypertension Study (TOMHS) are relevant to long-term adherence to various anti-hypertensive drug monotherapies. At 48 months, 82.5% and 77.8% of participants remained on amlodipine and acebutolol, respectively (both P < 0.01 compared with placebo). However, only 67.5%, 66.1% and 68.1%, respectively, of chlorthalidone, doxazosin and enalapril participants remained on these drugs as monotherapy at 48 months. Differential adherence to long-term anti-hypertensive drug therapy could translate into a greater risk of blood pressure-related complications and higher overall healthcare expenditures. Strategies to minimize the deleterious impact of therapeutic non-adherence with anti-hypertensive medications as well as the clinical and cost implications of the TOMHS data will be discussed.
Assuntos
Anti-Hipertensivos/uso terapêutico , Cooperação do Paciente , Anti-Hipertensivos/economia , Anti-Hipertensivos/metabolismo , Meia-Vida , Humanos , Ensaios Clínicos Controlados Aleatórios como Assunto , Resultado do TratamentoRESUMO
1 Doxazosin is a quinazoline derivative, related to prazosin, recently developed for the treatment of hypertension. 2 The intravenous administration of doxazosin (12 micrograms/kg) to six healthy normotensive subjects resulted in significant fall in erect blood pressure, with a corresponding increase in heart rate, but there were no significant changes in supine blood pressure or heart rate. 3 The changes in blood pressure and heart rate were maximal at 6 h after intravenous dosing. With prazosin the maximum effects occurred within the first hours. 4 Pressor response studies with phenylephrine confirmed that doxazosin is a relatively selective postsynaptic alpha-adrenoceptor antagonist. 5 The mean elimination half-life of doxazosin was 11 h. This compared with a T1/2 of 2.5 h for prazosin.