RESUMO
Abstract Dyslipidemia is an abnormal lipid profile associated with many common diseases, including coronary heart disease and atherosclerosis. Cholesteryl ester transfer protein (CETP) is a hydrophobic plasma glycoprotein that is responsible for the transfer of cholesteryl ester from high-density lipoprotein athero-protective particles to pro-atherogenic very low-density lipoprotein and low-density lipoprotein particles. The requirement for new CETP inhibitors, which block this process has driven our current work. Here, the synthesis as well as the ligand-based and structure-based design of seven oxoacetamido-benzamides 9a-g with CETP inhibitory activity is described. An in vitro study demonstrated that most of these compounds have appreciable CETP inhibitory activity. Compound 9g showed the highest inhibitory activity against CETP with an IC50 of 0.96 µM. Glide docking data for compounds 9a-g and torcetrapib provide evidence that they are accommodated in the CETP active site where hydrophobic interactions drive ligand/CETP complex formation. Furthermore, compounds 9a-g match the features of known CETP active inhibitors, providing a rationale for their high docking scores against the CETP binding domain. Therefore, these oxoacetamido-benzamides show potential for use as novel CETP inhibitors
Assuntos
Benzamidas/efeitos adversos , Dislipidemias/complicações , Proteínas de Transferência de Ésteres de Colesterol/antagonistas & inibidores , Técnicas In Vitro/métodos , Ésteres do Colesterol , Doença das Coronárias/patologia , Concentração Inibidora 50 , Lipoproteínas HDL/classificação , Lipoproteínas LDL/classificaçãoRESUMO
BACKGROUND: The functionality of high-density lipoproteins (HDL) is a better cardiovascular risk predictor than HDL concentrations. One of the key elements of HDL functionality is its apolipoprotein composition. Lecithin-cholesterol acyl transferase (LCAT) and cholesterol-ester transfer protein (CETP) are enzymes involved in HDL-mediated reverse cholesterol transport. This study assessed the concentration and activity of LCAT and CETP in HDL subspecies defined by their content of apolipoproteins E (apoE) and C-III (apoC-III) in humans. METHODS: Eighteen adults (ten women and eight men, mean age 55.6, BMI 26.9 Kg/m2, HbA1c 5.4%) were studied. HDL from each participant were isolated and divided into four subspecies containing respectively: No apoE and no apoC-III (E-C-), apoE but not apoC-III (E + C-), apoC-III but no apoE (E-C+) and both apoE and apoC-III (E + C+). The concentration and enzymatic activity of LCAT and CETP were measured within each HDL subspecies using immunoenzymatic and fluorometric methods. Additionally, the size distribution of HDL in each apolipoprotein-defined fraction was determined using non-denaturing electrophoresis and anti-apoA-I western blotting. RESULTS: HDL without apoE or apoC-III was the predominant HDL subtype. The size distribution of HDL was very similar in all the four apolipoprotein-defined subtypes. LCAT was most abundant in E-C- HDL (3.58 mg/mL, 59.6% of plasma LCAT mass), while HDL with apoE or apoC-III had much less LCAT (19.8, 12.2 and 8.37% of plasma LCAT respectively for E + C-, E-C+ and E + C+). LCAT mass was lower in E + C- HDL relative to E-C- HDL, but LCAT activity was similar in both fractions, signaling a greater activity-to-mass ratio associated with the presence of apoE. Both CETP mass and CETP activity showed only slight variations across HDL subspecies. There was an inverse correlation between plasma LCAT activity and concentrations of both E-C+ pre-beta HDL (r = - 0.55, P = 0.017) and E-C- alpha 1 HDL (r = - 0.49, P = 0.041). Conversely, there was a direct correlation between plasma CETP activity and concentrations of E-C+ alpha 1 HDL (r = 0.52, P = 0.025). CONCLUSIONS: The presence of apoE in small HDL is correlated with increased LCAT activity and esterification of plasma cholesterol. These results favor an interpretation that LCAT and apoE interact to enhance anti-atherogenic pathways of HDL.
Assuntos
Apolipoproteína C-III/análise , Apolipoproteínas E/análise , Proteínas de Transferência de Ésteres de Colesterol/análise , Colesterol/metabolismo , Lipoproteínas HDL/metabolismo , Fosfatidilcolina-Esterol O-Aciltransferase/análise , Adulto , Idoso , Proteínas de Transferência de Ésteres de Colesterol/sangue , Proteínas de Transferência de Ésteres de Colesterol/metabolismo , Ésteres do Colesterol/metabolismo , Feminino , Humanos , Lipoproteínas HDL/química , Lipoproteínas HDL/classificação , Masculino , Pessoa de Meia-Idade , Fosfatidilcolina-Esterol O-Aciltransferase/sangue , Fosfatidilcolina-Esterol O-Aciltransferase/metabolismoRESUMO
Standard lipoprotein measurements of triglycerides, total cholesterol, low-density lipoproteins (LDL), and high-density lipoproteins (HDL) fail to identify many lipoprotein abnormalities that contribute to cardiovascular heart diseases (CHD). Studies suggested that the presence of CHD is more strongly associated with the HDL subspecies than with total HDL cholesterol levels. The HDL particles can be collected in at least three subfractions, the HDL2b, HDL2a, and HDL3. More specifically, atherosclerosis is associated with low levels of HDL2. In this work, the optical spectroscopic properties of europium tetracycline (EuTc) complex in the presence of different HDL subspecies was studied. The results show that the europium spectroscopic properties in the EuTc complex are influenced by sizes and concentrations of subclasses. Eu3+ emission intensity and lifetime can discriminate the subfractions HDL3 and HDL2b.
Assuntos
Európio/química , Lipoproteínas HDL/sangue , Lipoproteínas HDL/química , Espectrometria de Fluorescência/métodos , Tetraciclinas/química , Humanos , Lipoproteínas HDL/classificaçãoRESUMO
OBJECTIVE: To establish normative data for lipoprotein subfractions using a novel ion mobility assay in healthy lean children and to compare their data with those of obese children preselected with normal glucose, blood pressure, and relatively normal lipids. STUDY DESIGN: Fasting blood samples in 162 children aged 7.0-18.9 years (75 lean [body mass index: 18.6 ± 6.6 kg/m(2)] and 87 obese [body mass index: 31.7 ± 5.4 kg/m(2)]) were analyzed. Correlation of lipoprotein subfractions with anthropometric and laboratory markers was performed. Principal component analysis was used to avoid using correlated variables. RESULTS: Normative data for lipid subfractions were obtained in healthy children. Lean children had higher high-density lipoprotein (HDL)-large (76%), HDL-small (13%), and HDL-total (27%) compared with obese (P < .01), and lower low-density lipoprotein (LDL)-medium (-30%, P < .01) and medium + small (-21%, P = .02) as well as LDL-total (-13%, P = .035). In both groups, the LDL component was higher in males and pubertal children (P < .01). Prepubertal children had a higher HDL component than pubertal ones (P < .004). Adjusting for sex and pubertal status LDL component was positively, and HDL component negatively, correlated with obesity (P < .004). CONCLUSIONS: Despite relatively normal triglycerides and cholesterol measured with standard assays at screening, ion mobility analysis showed significant differences in lipid and apolipoprotein subfractions between lean and obese children, even those prepubertal. Long-term, prospective follow-up may better characterize the predictability of lipid subfractions for future cardiovascular disease risk in children.
Assuntos
Lipoproteínas HDL/sangue , Lipoproteínas LDL/sangue , Obesidade/sangue , Adolescente , Biomarcadores/sangue , Glicemia/metabolismo , Pressão Sanguínea , Criança , Colesterol/sangue , Eletroforese/métodos , Feminino , Humanos , Lipoproteínas HDL/química , Lipoproteínas HDL/classificação , Lipoproteínas LDL/química , Lipoproteínas LDL/classificação , Modelos Logísticos , Masculino , Tamanho da Partícula , Análise de Componente Principal , Puberdade , Valores de Referência , Fatores Sexuais , Triglicerídeos/sangueRESUMO
BACKGROUND: Low cholesterol and phospholipid plasma levels of some high-density lipoprotein (HDL) subclasses have been described in children with metabolic syndrome. Scavenger receptor class B type I (SR-BI) has been proposed to be at the origin of such HDL alterations because of its key role on cholesteryl esters-HDL metabolism. However, the possible contribution of SR-BI has not been specifically explored in this kind of patients. METHODS: Plasma lipid concentrations of HDL subclasses, i.e., triglycerides (TG), phosphatidylcholine (Ph), free cholesterol (FC), and total cholesterol (TC), were determined by enzymatic staining on polyacrylamide gradient gels (PAGE) in 39 pediatric patients with metabolic syndrome and 65 children as controls. Cholesteryl esters were estimated by the difference between TC and FC. Proteins of HDL subclasses were also stained for the assessment of the relative size distribution of HDL. For statistical analysis, the study population was grouped by Srb1 +1050C-->T polymorphism (rs5888) as carriers or noncarriers of the T allele, and data were corrected by metabolic syndrome status. RESULTS: The Srb1 +1050T allele was associated with metabolic syndrome [odds ratio (OR)=2.18 (1.12-4.22), P=0.02]. Plasma TG corresponding to HDL3a, as well as the relative proportion of this HDL subclass, were slightly higher in carriers of the T allele as compared to CC homozygous subjects. Cholesteryl esters plasma concentrations of all HDL subclasses were comparable between T allele carriers and noncarriers after correction by metabolic syndrome status. CONCLUSIONS: Srb1 +1050T was associated with metabolic syndrome, but T carrier subjects did not show important differences concerning HDL subclasses as compared to noncarriers.
Assuntos
Ésteres do Colesterol/sangue , Lipoproteínas HDL/sangue , Síndrome Metabólica/sangue , Síndrome Metabólica/genética , Polimorfismo de Nucleotídeo Único , Receptores Depuradores Classe B/genética , Adolescente , Idade de Início , Alelos , Estudos de Casos e Controles , Criança , Ésteres do Colesterol/classificação , Feminino , Frequência do Gene , Predisposição Genética para Doença , Humanos , Lipoproteínas HDL/classificação , Masculino , Síndrome Metabólica/epidemiologia , Concentração Osmolar , Polimorfismo de Nucleotídeo Único/fisiologiaRESUMO
BACKGROUND: Small HDL particles have emerged as significant predictors of incident type 2 diabetes mellitus (T2DM) in adults with impaired glucose tolerance (IGT). However, no previous study has investigated HDL size in pediatric subjects with these clinical conditions. METHODS: We studied the HDL size distribution by native polyacrilamide gradient gel electrophoresis in 106 overweight children, 47 with T2DM, 43 with normal glucose tolerance (NGT), 16 with IGT, and 39 healthy weight controls. RESULTS: Diabetic children had significantly lower proportions of HDL2b and HDL2a, and higher proportions of HDL3b and HDL3c than the other 3 groups. Overweight subjects showed HDL size distributions similar to those of controls. However, insulin-resistant children had lower proportions of HDL2b, and HDL2a, and higher proportions of HDL3b when compared with the insulin-sensitive overweight subjects. Multiple linear regression analyses showed that homeostasis model assessment correlated inversely with HDL2b and HDL2a, and directly with HDL3b, while BMI was independently associated only with HDL3a. CONCLUSIONS: This study showed that HDL size distribution was shifted toward smaller particles in T2DM pediatric patients and in overweight children with insulin resistance, independent of their glucose tolerance status. Insulin resistance was the main factor associated with these HDL size abnormalities. This parameter could be useful as an early risk marker of incident diabetes and, probably, of coronary heart disease.