Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 26
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
Nutrients ; 10(7)2018 Jul 19.
Artigo em Inglês | MEDLINE | ID: mdl-30029523

RESUMO

(1) Background: Arteriosclerosis is associated with high levels of low-density lipoprotein (LDL) cholesterol. O-methylated catechins in "Benifuuki" green tea are expected to reduce cholesterol levels, although there is limited research regarding this topic; (2) Methods: This trial evaluated 159 healthy volunteers who were randomized to receive ice cream containing a high-dose of "Benifuuki" extract including 676 mg of catechins (group H), a low-dose of "Benifuuki" extract including 322 mg of catechins (group L), or no "Benifuuki" extract (group C). Each group consumed ice cream (with or without extract) daily for 12 weeks, and their lipid-related parameters were compared; (3) Results: A significant reduction in the level of lectin-like oxidized LDL receptor-1 ligand containing ApoB (LAB) was detected in group H, compared to groups L and C. No significant differences between the three groups were detected in their levels of total cholesterol, triglycerides, and LDL cholesterol; (4) Conclusions: "Benifuuki" extract containing O-methylated catechins may help prevent arteriosclerosis.


Assuntos
Apolipoproteína B-100/antagonistas & inibidores , Camellia sinensis/química , Suplementos Nutricionais , Hiperlipidemias/prevenção & controle , Hipolipemiantes/administração & dosagem , Extratos Vegetais/administração & dosagem , Receptores Depuradores Classe E/metabolismo , Idoso , Apolipoproteína B-100/sangue , Biomarcadores/sangue , Catecóis/administração & dosagem , Catecóis/uso terapêutico , Método Duplo-Cego , Feminino , Manipulação de Alimentos , Preferências Alimentares , Humanos , Hiperlipidemias/sangue , Hipolipemiantes/uso terapêutico , Sorvetes , Análise de Intenção de Tratamento , Japão , Ligantes , Masculino , Pessoa de Meia-Idade , Oxirredução , Extratos Vegetais/uso terapêutico , Folhas de Planta/química
2.
Biosci Biotechnol Biochem ; 81(8): 1569-1575, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28463548

RESUMO

Oxidation of low-density lipoprotein (LDL) by reactive oxygen species (ROS) and reactive nitrogen species (RNS) has been suggested to be involved in the onset of atherosclerosis. Oolong tea contains unique polyphenols including oolonghomobisflavan A (OFA). In this study, the effects of OFA on LDL oxidation by ROS and RNS were investigated in vitro. OFA suppressed formation of cholesterol ester hydroperoxides in LDL oxidized by peroxyl radical and peroxynitrite, and formation of thiobarbituric acid reactive substances in LDL oxidized by Cu2+. In addition, OFA inhibited fragmentation, carbonylation, and nitration of apolipoprotein B-100 (apo B-100) in the oxidized LDL, in which heparin-binding activity of apo B-100 was protected by OFA. Our results suggest that OFA exhibits antioxidant activity against both lipid peroxidation and oxidative modification of apo B-100 in LDL oxidized by ROS and RNS. Polyphenols in oolong tea may prevent atherosclerosis by reducing oxidative stress.


Assuntos
Camellia sinensis/química , Flavonoides/química , Lipoproteínas LDL/antagonistas & inibidores , Polifenóis/química , Apolipoproteína B-100/antagonistas & inibidores , Cátions Bivalentes , Ésteres do Colesterol/antagonistas & inibidores , Cobre/química , Flavonoides/isolamento & purificação , Heparina/química , Humanos , Cinética , Peroxidação de Lipídeos , Oxirredução , Peróxidos/antagonistas & inibidores , Ácido Peroxinitroso/antagonistas & inibidores , Extratos Vegetais/química , Polifenóis/isolamento & purificação , Ligação Proteica , Espécies Reativas de Nitrogênio/antagonistas & inibidores , Espécies Reativas de Oxigênio/antagonistas & inibidores , Tiobarbitúricos/antagonistas & inibidores
3.
World J Gastroenterol ; 22(45): 9954-9965, 2016 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-28018102

RESUMO

AIM: To characterize the role of apolipoprotein B100 (apoB100) in hepatitis C viral (HCV) infection. METHODS: In this study, we utilize a gene editing tool, transcription activator-like effector nucleases (TALENs), to generate human hepatoma cells with a stable genetic deletion of APOB to assess of apoB in HCV. Using infectious cell culture-competent HCV, viral pseudoparticles, replicon models, and lipidomic analysis we determined the contribution of apoB to each step of the viral lifecycle. We further studied the effect of mipomersen, an FDA-approved antisense inhibitor of apoB100, on HCV using in vitro cell-culture competent HCV and determined its impact on viral infectivity with the TCID50 method. RESULTS: We found that apoB100 is indispensable for HCV infection. Using the JFH-1 fully infectious cell-culture competent virus in Huh 7 hepatoma cells with TALEN-mediated gene deletion of apoB (APOB KO), we found a significant reduction in HCV RNA and protein levels following infection. Pseudoparticle and replicon models demonstrated that apoB did not play a role in HCV entry or replication. However, the virus produced by APOB KO cells had significantly diminished infectivity as measured by the TCID-50 method compared to wild-type virus. Lipidomic analysis demonstrated that these virions have a fundamentally altered lipidome, with complete depletion of cholesterol esters. We further demonstrate that inhibition of apoB using mipomersen, an FDA-approved anti-sense oligonucleotide, results in a potent anti-HCV effect and significantly reduces the infectivity of the virus. CONCLUSION: ApoB is required for the generation of fully infectious HCV virions, and inhibition of apoB with mipomersen blocks HCV. Targeting lipid metabolic pathways to impair viral infectivity represents a novel host targeted strategy to inhibit HCV.


Assuntos
Apolipoproteína B-100/genética , Hepatite C/genética , Hepatócitos/metabolismo , RNA Viral/metabolismo , Internalização do Vírus , Apolipoproteína B-100/antagonistas & inibidores , Linhagem Celular , Técnicas de Inativação de Genes , Hepacivirus , Hepatite C/virologia , Hepatócitos/efeitos dos fármacos , Hepatócitos/virologia , Humanos , Técnicas In Vitro , Oligodesoxirribonucleotídeos Antissenso/farmacologia , Oligonucleotídeos/farmacologia , Proteínas Virais/metabolismo , Internalização do Vírus/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos
4.
Sci Transl Med ; 8(323): 323ra12, 2016 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-26819195

RESUMO

Mipomersen is a 20mer antisense oligonucleotide (ASO) that inhibits apolipoprotein B (apoB) synthesis; its low-density lipoprotein (LDL)-lowering effects should therefore result from reduced secretion of very-low-density lipoprotein (VLDL). We enrolled 17 healthy volunteers who received placebo injections weekly for 3 weeks followed by mipomersen weekly for 7 to 9 weeks. Stable isotopes were used after each treatment to determine fractional catabolic rates and production rates of apoB in VLDL, IDL (intermediate-density lipoprotein), and LDL, and of triglycerides in VLDL. Mipomersen significantly reduced apoB in VLDL, IDL, and LDL, which was associated with increases in fractional catabolic rates of VLDL and LDL apoB and reductions in production rates of IDL and LDL apoB. Unexpectedly, the production rates of VLDL apoB and VLDL triglycerides were unaffected. Small interfering RNA-mediated knockdown of apoB expression in human liver cells demonstrated preservation of apoB secretion across a range of apoB synthesis. Titrated ASO knockdown of apoB mRNA in chow-fed mice preserved both apoB and triglyceride secretion. In contrast, titrated ASO knockdown of apoB mRNA in high-fat-fed mice resulted in stepwise reductions in both apoB and triglyceride secretion. Mipomersen lowered all apoB lipoproteins without reducing the production rate of either VLDL apoB or triglyceride. Our human data are consistent with long-standing models of posttranscriptional and posttranslational regulation of apoB secretion and are supported by in vitro and in vivo experiments. Targeting apoB synthesis may lower levels of apoB lipoproteins without necessarily reducing VLDL secretion, thereby lowering the risk of steatosis associated with this therapeutic strategy.


Assuntos
Apolipoproteína B-100/antagonistas & inibidores , Fígado/metabolismo , Adolescente , Adulto , Idoso , Animais , Apolipoproteínas B/genética , Feminino , Voluntários Saudáveis , Células Hep G2 , Humanos , Lipoproteínas IDL/sangue , Lipoproteínas LDL/sangue , Lipoproteínas VLDL/sangue , Masculino , Camundongos , Pessoa de Meia-Idade , Oligonucleotídeos/química , Oligonucleotídeos Antissenso/química , RNA Interferente Pequeno/metabolismo , Triglicerídeos/sangue , Triglicerídeos/metabolismo , Adulto Jovem
5.
J Clin Lipidol ; 9(2): 201-9, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25911076

RESUMO

BACKGROUND: Because of variability in lipoprotein cholesterol content, low-density lipoprotein (LDL) cholesterol frequently underrepresents or overrepresents the number of LDL particles. Mipomersen is an antisense oligonucleotide that reduces hepatic production of apolipoprotein B-100, the sole apolipoprotein of LDL. OBJECTIVE: To characterize the effects of mipomersen on lipoprotein particle numbers as well as subclass distribution using nuclear magnetic resonance (NMR) spectroscopy. METHODS: We compared the tertiary results for the direct measurement of LDL particle numbers by NMR among 4 placebo-controlled, phase 3 studies of mipomersen that had similar study designs but different patient populations: homozygous familial hypercholesterolemia (HoFH), severe hypercholesterolemia, heterozygous familial hypercholesterolemia with established coronary artery disease, or hypercholesterolemia with high risk for coronary heart disease (HC-CHD). RESULTS: HoFH patients had the highest median total LDL particles at baseline compared with HC-CHD patients, who had the lowest. At baseline, the HoFH population uniquely had a greater mean percentage of large LDL particles (placebo, 60.2%; mipomersen, 54.9%) compared with small LDL particles (placebo, 33.1%; mipomersen, 38.9%). In all 4 studies, mipomersen was associated with greater reductions from baseline in the concentrations of small LDL particles compared with those of large LDL particles, and both total LDL particles and small LDL particles were statistically significantly reduced. CONCLUSIONS: Mipomersen consistently reduced all LDL particle numbers and preferentially reduced the concentration of small LDL particles in all 4 phase 3 studies.


Assuntos
Doença da Artéria Coronariana/sangue , Doença da Artéria Coronariana/tratamento farmacológico , Hipercolesterolemia/tratamento farmacológico , Oligonucleotídeos/administração & dosagem , Adulto , Apolipoproteína B-100/antagonistas & inibidores , Apolipoproteína B-100/genética , HDL-Colesterol/sangue , LDL-Colesterol/sangue , Doença da Artéria Coronariana/patologia , Feminino , Humanos , Hipercolesterolemia/sangue , Hipercolesterolemia/patologia , Masculino , Pessoa de Meia-Idade , Ressonância Magnética Nuclear Biomolecular
6.
Colloids Surf B Biointerfaces ; 129: 71-8, 2015 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-25829129

RESUMO

BACKGROUND: Anionic nanoliposomes can interact with serum lipoproteins and regulate lipid metabolism through several mechanisms. This study aimed to evaluate the lipid-modifying effects of anionic immunoliposomes targeted against apoB, an important component of atherogenic lipoproteins. METHODS: Two sets of nanoliposomes (20mM) were prepared with low (including soy phosphatidylcholine [SPC] and egg phosphatidylglycerol [EPG]) and high (including hydrogenated soy phosphatidylcholine [HSPC] and distearoyl phosphatidylglycerol [DSPG]) phase transition temperature values without cholesterol. In each set, the anionic phospholipid (EPG or DSPG) constituted 75% of total phospholipid content. Immunoliposomes were prepared by conjugating a monoclonal antibody against apoB-100 to the liposomal surface using a post-insertion technique. Fluorescently-labeled immunoliposomes were assessed for their uptake by J774.A1 macrophages. Lipid-modifying effects of immunoliposomes were tested at different doses (50, 100 or 200µmole/g weight) using a tyloxapol-induced hyperlipidemic mouse model. Blood sampling was performed 1h after the injection of each immunoliposomal formulation. RESULTS: ApoB-targeted HSPC/DSPG and SPC/EPG nanoliposomes were both taken up by cultured macrophages but the uptake rate was higher with the former formulation. Both immunoliposomal formulations significantly reduced serum LDL-cholesterol concentrations of hyperlipidemic animals at all tested doses (p<0.001) and this effect lasted for at least 48h. Significant reductions of serum levels of apoB, non-HDL-C, total cholesterol and triglycerides, and elevations of HDL-C levels were also observed. CONCLUSION: Intravenous injection of a single dose of apoB-targeted anionic nanoliposomes improves serum lipid profile parameters. These findings might have implications for the treatment of patients with severe dyslipidemias or statin intolerance.


Assuntos
Anticorpos Monoclonais/farmacologia , Apolipoproteína B-100/antagonistas & inibidores , Sistemas de Liberação de Medicamentos , Dislipidemias/tratamento farmacológico , Lipossomos , Nanopartículas/química , Animais , Anticorpos Monoclonais/química , Apolipoproteína B-100/imunologia , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Colesterol/química , Colesterol/metabolismo , Dislipidemias/metabolismo , Dislipidemias/patologia , Lipoproteínas/química , Lipoproteínas/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Masculino , Camundongos , Fosfolipídeos/química , Fosfolipídeos/metabolismo
7.
Autoimmunity ; 48(3): 152-60, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25683179

RESUMO

Atherosclerosis is a chronic inflammatory disease of the artery wall. Adaptive immunity plays a key role in the pathogenesis of atherosclerosis. Recently, modulation of the immune response against atherosclerotic plaque antigen(s) has attracted attention as a potentially preventive and therapeutic approach. Here, we review a series of studies on immunization with various antigens targeting treatment and prevention of atherosclerosis. Atherosclerosis-related antigens include oxidized low-density lipoprotein (LDL), apolipoprotein B-100 (ApoB-100) and heat shock protein (HSP) 60/65. Accumulating evidence supports the idea that immunization with these antigenic proteins or peptides may reduce atherosclerosis. In this review, we discuss the current status of immunization studies and possible associated mechanisms of atheroprotection.


Assuntos
Anticorpos/farmacologia , Aterosclerose/prevenção & controle , Placa Aterosclerótica/prevenção & controle , Vacinação , Vacinas/imunologia , Imunidade Adaptativa/efeitos dos fármacos , Animais , Apolipoproteína B-100/antagonistas & inibidores , Apolipoproteína B-100/genética , Apolipoproteína B-100/imunologia , Artérias/efeitos dos fármacos , Artérias/imunologia , Artérias/patologia , Aterosclerose/imunologia , Aterosclerose/patologia , Autoantígenos/genética , Autoantígenos/imunologia , Expressão Gênica , Proteínas de Choque Térmico/antagonistas & inibidores , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/imunologia , Humanos , Imunidade Humoral/efeitos dos fármacos , Lipoproteínas LDL/antagonistas & inibidores , Lipoproteínas LDL/genética , Lipoproteínas LDL/imunologia , Camundongos , Peptídeos/administração & dosagem , Peptídeos/genética , Peptídeos/imunologia , Placa Aterosclerótica/imunologia , Placa Aterosclerótica/patologia , Vacinas/administração & dosagem , Vacinas/genética
8.
Nucleic Acid Ther ; 24(4): 283-90, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24758560

RESUMO

The multiple-turnover ability of a series of locked nucleic acid (LNA)-based antisense oligonucleotides (AONs) in the RNase H-mediated scission reaction was estimated using a newly developed cell-free reaction system. We determined the initial reaction rates of AONs under multiple-turnover conditions and found that among 24 AONs tested, AONs with melting temperatures (Tm) of 40°C-60°C efficiently elicit multiple rounds of RNA scission. On the other hand, by measuring Tm with two 10-mer RNAs partially complementary to AONs as models of cleaved 5' and 3' fragments of mRNA, we found that AONs require adequate binding affinity for efficient turnover activities. We further demonstrated that the efficacy of a set of 13-mer AONs in mice correlated with their turnover efficiency, indicating that the intracellular situation where AONs function is similar to multiple-turnover conditions. Our methodology and findings may provide an opportunity to shed light on a previously unknown antisense mechanism, leading to further improvement of the activity and safety profiles of AONs.


Assuntos
Apolipoproteína B-100/genética , Oligonucleotídeos Antissenso/química , Oligonucleotídeos/química , RNA Mensageiro/genética , Ribonuclease H/química , Animais , Apolipoproteína B-100/antagonistas & inibidores , Apolipoproteína B-100/metabolismo , Sistema Livre de Células , Expressão Gênica , Meia-Vida , Injeções Subcutâneas , Fígado/química , Fígado/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Oligonucleotídeos/metabolismo , Oligonucleotídeos/farmacocinética , Oligonucleotídeos Antissenso/metabolismo , Oligonucleotídeos Antissenso/farmacocinética , Estabilidade de RNA , RNA Mensageiro/antagonistas & inibidores , RNA Mensageiro/metabolismo , Ribonuclease H/metabolismo , Temperatura de Transição
9.
Eur Heart J ; 34(42): 3251-8, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23966311
10.
J Cardiovasc Pharmacol Ther ; 18(3): 199-210, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23580658

RESUMO

Dyslipidemia is a major risk factor for the development of coronary artery disease, a leading cause of morbidity and mortality worldwide. Lowering low-density lipoprotein (LDL) has significantly reduced the risk of death and other major cardiovascular events, and statins remain the therapy of choice. However, as some patients are limited by the side effects of statins, cannot achieve their target LDL on statin therapy, or have other abnormalities in their lipid profile, alternative agents are being developed. In this review, we highlight the major classes of novel nonstatin LDL-lowering agents that are currently in various stages of development. Although many hold great promise, the results of large Phase III trials will be needed to definitely establish the efficacy, safety, and clinical utility of these agents in the general population.


Assuntos
Drogas em Investigação/uso terapêutico , Dislipidemias/tratamento farmacológico , Hipolipemiantes/uso terapêutico , Terapia de Alvo Molecular , Animais , Apolipoproteína B-100/antagonistas & inibidores , Transporte Biológico/efeitos dos fármacos , Proteínas de Transporte/antagonistas & inibidores , Proteínas de Transferência de Ésteres de Colesterol/antagonistas & inibidores , Drogas em Investigação/efeitos adversos , Drogas em Investigação/farmacologia , Dislipidemias/sangue , Dislipidemias/metabolismo , Humanos , Hipolipemiantes/efeitos adversos , Hipolipemiantes/farmacologia , Lipoproteínas LDL/antagonistas & inibidores , Lipoproteínas LDL/sangue , Lipoproteínas LDL/metabolismo , Pró-Proteína Convertases/antagonistas & inibidores , Pró-Proteína Convertases/metabolismo , Inibidores de Proteases/efeitos adversos , Inibidores de Proteases/farmacologia , Inibidores de Proteases/uso terapêutico
11.
Eur Heart J ; 34(13): 962-71, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23416791

RESUMO

Familial hypercholesterolaemia (FH) is an autosomal dominant genetic disorder, associated with elevated levels of low-density lipoprotein-cholesterol (LDL-C), which can lead to premature cardiovascular disease. Early diagnosis of FH is important to prevent morbidity and mortality. Familial hypercholesterolaemia is usually diagnosed using clinical characteristics, such as family history, and cholesterol levels; however, genetic testing may provide a definite diagnosis of FH by detecting a pathological mutation. Current guidelines highlight the importance of reducing LDL-C levels in patients with FH. Statins are the current standard treatment for the majority of these patients, and have been shown to be effective in reducing the incidence of cardiovascular heart disease in patients with FH. Nevertheless, many FH patients do not achieve their target LDL-C levels; as such, new treatment options are required to decrease LDL-C levels beyond those currently achieved. There are currently several new classes of pharmacotherapy under investigation to control LDL-C levels. These include agents which modify LDL-C production, such as inhibitors of apolipoprotein B, or those which affect LDL-C catabolism, such as inhibition of pro-protein convertase subtilisin/kexin 9, a protein which is responsible for the degradation of the LDL receptor. Therapies which raise high-density lipoprotein cholesterol are also being evaluated. In this article, we consider the diagnosis of FH and the goals of therapy and review the current and potential future treatment options for patients with FH.


Assuntos
Doenças Cardiovasculares/etiologia , Hiperlipoproteinemia Tipo II/tratamento farmacológico , Anticolesterolemiantes/uso terapêutico , Apolipoproteína B-100/antagonistas & inibidores , Apolipoproteína B-100/genética , Doenças Cardiovasculares/prevenção & controle , Colesterol/metabolismo , LDL-Colesterol/efeitos dos fármacos , LDL-Colesterol/metabolismo , Diagnóstico Precoce , Testes Genéticos , Terapia Genética/métodos , Heterozigoto , Homozigoto , Humanos , Hiperlipoproteinemia Tipo II/diagnóstico , Hiperlipoproteinemia Tipo II/genética , Lipoproteínas/antagonistas & inibidores , Lipoproteínas/metabolismo , Pró-Proteína Convertase 9 , Pró-Proteína Convertases/genética , Receptores de LDL/genética , Fatores de Risco , Serina Endopeptidases/genética
12.
Drugs ; 72(11): 1445-55, 2012 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-22799743

RESUMO

High levels of low-density lipoprotein cholesterol (LDL-C) and lipoprotein(a) [Lp(a)] are associated with early morbidity and mortality caused by cardiovascular disease (CVD). There are hints that a reduction of LDL-C levels beyond currently advocated targets, and the use of drugs that also have Lp(a)-lowering potential, could provide further clinical benefit. Today, LDL apheresis is the only available treatment option to achieve further lowering of apolipoprotein-B (apo-B)-containing lipoproteins, especially Lp(a). Mipomersen is currently being studied in patients with mild to severe hypercholesterolaemia as add-on therapy to other lipid-lowering therapy, as monotherapy in patients who are intolerant of HMG-CoA reductase inhibitors (statins) and who are at high risk for CVD. Patients affected by homozygous or heterozygous familial hypercholesterolaemia (FH), which are inherited autosomal co-dominant disorders characterized by a marked elevation of serum LDL-C concentration, remain a clinical challenge, especially when their CVD risk is aggravated by additionally elevated Lp(a) levels. Mipomersen is a 20-mer oligonucleotide [2'-O-(2-methoxy) ethyl-modified oligonucleotide], a second-generation antisense oligonucleotide (AOS), complementary to the coding region for human-specific apo-B-100 messenger RNA (mRNA). Mipomersen inhibits apo-B-100 synthesis and is consequently a new treatment strategy to lower apo-B-containing lipoproteins like LDL-C and Lp(a) in patients at high risk for CVD not on target or intolerant to statins. This article focuses on mipomersen and gives an overview of the current status of mipomersen as a promising treatment option. Recent studies have shown a decrease in LDL-C levels of 22-42.2% and in Lp(a) of 19.6-31.1% from baseline, depending on study design. Dose-dependent reductions of very low-density lipoprotein cholesterol (VLDL-C) and triglyceride levels have also been observed. Although the short-term efficacy and safety of mipomersen have been proven, side effects like injection-site reactions (up to 90-100%), increased liver enzymes, cephalgias, nasopharyngitis, myalgia, nausea and fatigue must be mentioned and critically discussed. Furthermore, we need more data on the long-term side effects, especially regarding the long-term potential for hepatic steatosis. Data on cardiovascular outcomes with mipomersen are also not yet available.


Assuntos
Anticolesterolemiantes/uso terapêutico , Hiperlipoproteinemia Tipo II/tratamento farmacológico , Oligonucleotídeos/uso terapêutico , Animais , Anticolesterolemiantes/efeitos adversos , Apolipoproteína B-100/antagonistas & inibidores , Apolipoproteína B-100/sangue , Apolipoproteína B-100/metabolismo , LDL-Colesterol/sangue , LDL-Colesterol/metabolismo , VLDL-Colesterol/sangue , VLDL-Colesterol/metabolismo , Ensaios Clínicos Fase II como Assunto , Ensaios Clínicos Fase III como Assunto , Método Duplo-Cego , Avaliação Pré-Clínica de Medicamentos , Humanos , Hiperlipoproteinemia Tipo II/sangue , Hiperlipoproteinemia Tipo II/metabolismo , Lipoproteína(a)/sangue , Lipoproteína(a)/metabolismo , Oligonucleotídeos/efeitos adversos , Ensaios Clínicos Controlados Aleatórios como Assunto , Triglicerídeos/sangue , Triglicerídeos/metabolismo
13.
BMC Biotechnol ; 12: 42, 2012 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-22827812

RESUMO

BACKGROUND: Controlling and limiting the expression of short hairpin RNA (shRNA) by using constitutive or tissue-specific polymerase II (pol II) expression can be a promising strategy to avoid RNAi toxicity. However, to date detailed studies on requirements for effective pol II shRNA expression and processing are not available. We investigated the optimal structural configuration of shRNA molecules, namely: hairpin location, stem length and termination signal required for effective pol II expression and compared it with an alternative strategy of avoiding toxicity by using artificial microRNA (miRNA) scaffolds. RESULTS: Highly effective shRNAs targeting luciferase (shLuc) or Apolipoprotein B100 (shApoB1 and shApoB2) were placed under the control of the pol II CMV promoter and expressed at +5 or +6 nucleotides (nt) with reference to the transcription start site (TSS). Different transcription termination signals (TTS), namely minimal polyadenylation (pA), poly T (T5) and U1 were also used. All pol II- expressed shRNA variants induced mild inhibition of Luciferase reporters carrying specific targets and none of them showed comparable efficacy to their polymerase III-expressed H1-shRNA controls, regardless of hairpin position and termination signal used. Extending hairpin stem length from 20 basepairs (bp) to 21, 25 or 29 bp yielded only slight improvement in the overall efficacy. When shLuc, shApoB1 and shApoB2 were placed in an artificial miRNA scaffold, two out of three were as potent as the H1-shRNA controls. Quantification of small interfering RNA (siRNA) molecules showed that the artificial miRNA constructs expressed less molecules than H1-shRNAs and that CMV-shRNA expressed the lowest amount of siRNA molecules suggesting that RNAi processing in this case is least effective. Furthermore, CMV-miApoB1 and CMV-miApoB2 were as effective as the corresponding H1-shApoB1 and H1-shApoB2 in inhibiting endogenous ApoB mRNA. CONCLUSION: Our results demonstrate that artificial miRNA have a better efficacy profile than shRNA expressed either from H1 or CMV promoter and will be used in the future for RNAi therapeutic development.


Assuntos
Apolipoproteína B-100/antagonistas & inibidores , Luciferases/antagonistas & inibidores , MicroRNAs/metabolismo , RNA Polimerase II/metabolismo , RNA Interferente Pequeno/metabolismo , Animais , Apolipoproteína B-100/genética , Apolipoproteína B-100/metabolismo , Linhagem Celular , Citomegalovirus/genética , Células HEK293 , Humanos , Luciferases/genética , Luciferases/metabolismo , Regiões Promotoras Genéticas , Interferência de RNA , RNA Polimerase II/genética , RNA Polimerase III/genética , RNA Polimerase III/metabolismo , Sítio de Iniciação de Transcrição
14.
J Med Chem ; 55(13): 6162-75, 2012 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-22650305

RESUMO

A series of diphenylpyridylethanamine (DPPE) derivatives was identified exhibiting potent CETP inhibition. Replacing the labile ester functionality in the initial lead 7 generated a series of amides and ureas. Further optimization of the DPPE series for potency resulted in the discovery of cyclopentylurea 15d, which demonstrated a reduction in cholesterol ester transfer activity (48% of predose level) in hCETP/apoB-100 dual transgenic mice. The PK profile of 15d was suboptimal, and further optimization of the N-terminus resulted in the discovery of amide 20 with an improved PK profile and robust efficacy in transgenic hCETP/apoB-100 mice and in hamsters. Compound 20 demonstrated no significant changes in either mean arterial blood pressure or heart rate in telemeterized rats despite sustained high exposures.


Assuntos
Anticolesterolemiantes/química , Anticolesterolemiantes/farmacologia , Proteínas de Transferência de Ésteres de Colesterol/antagonistas & inibidores , Piridinas/química , Piridinas/farmacologia , Estilbenos/química , Estilbenos/farmacologia , Animais , Anticolesterolemiantes/síntese química , Apolipoproteína B-100/antagonistas & inibidores , Apolipoproteína B-100/metabolismo , Pressão Sanguínea/efeitos dos fármacos , Proteínas de Transferência de Ésteres de Colesterol/metabolismo , Doença das Coronárias/tratamento farmacológico , Cricetinae , Descoberta de Drogas , Frequência Cardíaca/efeitos dos fármacos , Humanos , Concentração Inibidora 50 , Masculino , Camundongos , Camundongos Transgênicos , Estrutura Molecular , Piridinas/síntese química , Ratos , Estilbenos/síntese química
15.
Eur Heart J ; 33(9): 1142-9, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22507979

RESUMO

AIMS: A randomized, double-blind, placebo-controlled study was conducted to investigate the safety and efficacy of mipomersen, an apolipoprotein B-100 (apoB) synthesis inhibitor, in patients who are statin intolerant and at high risk for cardiovascular disease (CVD). METHODS AND RESULTS: Thirty-three subjects, not receiving statin therapy because of statin intolerance, received a weekly subcutaneous dose of 200 mg mipomersen or placebo (2:1 randomization) for 26 weeks. The primary endpoint was per cent change in LDL cholesterol (LDL-c) from the baseline to Week 28. The other efficacy endpoints were per cent change in apoB and lipoprotein a [Lp(a)]. Safety was determined using the incidence of treatment-emergent adverse events (AEs) and clinical laboratory evaluations. After 26 weeks of mipomersen administration, LDL-c was reduced by 47 ± 18% (P < 0.001 vs. placebo). apoB and Lp(a) were also significantly reduced by 46 and 27%, respectively (P < 0.001 vs. placebo). Four mipomersen (19%) and two placebo subjects (17%) discontinued dosing prematurely due to AEs. Persistent liver transaminase increases ≥ 3× the upper limit of normal were observed in seven (33%) subjects assigned to mipomersen. In selected subjects, liver fat content was assessed, during and after treatment, using magnetic resonance spectroscopy. Liver fat content in these patients ranged from 0.8 to 47.3%. Liver needle biopsy was performed in two of these subjects, confirming hepatic steatosis with minimal inflammation or fibrosis. CONCLUSION: The present data suggest that mipomersen is a potential therapeutic option in statin-intolerant patients at high risk for CVD. The long-term follow-up of liver safety is required. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov identifier: NCT00707746.


Assuntos
Anticolesterolemiantes/uso terapêutico , Apolipoproteína B-100/antagonistas & inibidores , LDL-Colesterol/metabolismo , Inibidores de Hidroximetilglutaril-CoA Redutases/efeitos adversos , Hipercolesterolemia/tratamento farmacológico , Oligonucleotídeos/uso terapêutico , Adulto , Idoso , Alanina Transaminase/metabolismo , Doenças Cardiovasculares/enzimologia , Doenças Cardiovasculares/prevenção & controle , Método Duplo-Cego , Feminino , Humanos , Hipercolesterolemia/enzimologia , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Resultado do Tratamento
17.
J Proteome Res ; 11(5): 2786-97, 2012 May 04.
Artigo em Inglês | MEDLINE | ID: mdl-22443280

RESUMO

Hepatic fat export occurs by apolipoprotein B-100-containing lipoprotein production, whereas impaired production leads to liver steatosis. Hepatitis C virus (HCV) infection is associated to dysregulation of apoB-100 secretion and steatosis; however, the molecular mechanism by which HCV affects the apoB-100 secretion is not understood. Here, combining quantitative proteomics and computational biology, we propose ferritin heavy chain (Fth) as being the cellular determinant of apoB-100 production inhibition. By means of molecular analyses, we found that HCV nonstructural proteins and NS5A appear to be sufficient for inducing Fth up-regulation. Fth in turn was found to inhibit apoB-100 secretion leading to increased intracellular degradation via proteasome. Notably, intracellular Fth down-regulation by siRNA restores apoB-100 secretion. The inverse correlation between ferritin and plasma apoB-100 concentrations was also found in JFH-1 HCV cell culture systems (HCVcc) and HCV-infected patients. Finally, Fth expression was found to be required for robust HCV infection. These observations provide a further molecular explanation for the onset of liver steatosis and allow for hypothesizing on new therapeutic and antiviral strategies.


Assuntos
Apoferritinas/metabolismo , Apolipoproteína B-100/antagonistas & inibidores , Regulação Viral da Expressão Gênica , Hepacivirus/patogenicidade , Apolipoproteína B-100/sangue , Linhagem Celular Tumoral , Biologia Computacional , Fígado Gorduroso/metabolismo , Fígado Gorduroso/patologia , Fígado Gorduroso/virologia , Hepacivirus/genética , Hepacivirus/metabolismo , Hepatite C/patologia , Hepatite C/virologia , Hepatócitos/metabolismo , Hepatócitos/patologia , Hepatócitos/virologia , Interações Hospedeiro-Patógeno , Humanos , Marcação por Isótopo , Complexo de Endopeptidases do Proteassoma/metabolismo , Mapas de Interação de Proteínas , Proteólise , Proteômica/métodos , Transfecção , Proteínas não Estruturais Virais/genética , Proteínas não Estruturais Virais/metabolismo
18.
Cardiol Rev ; 20(2): 90-5, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22293857

RESUMO

Mipomersen is an antisense oligonucleotide inhibitor of apolipoprotein (apo) B-100 currently in phase 3 of development for the treatment of hyperlipidemia in patients with a high risk for cardiovascular disease. The drug acts by inhibiting the production of apoB-100, which is the structural core for all atherogenic lipids, including low-density lipoprotein cholesterol (LDL-C). The agent has been shown to produce significant reductions in LDL-C from baseline values compared with placebos. Clinical trials have demonstrated that mipomersen reduces LDL-C up to 44% in patients with familial hypercholesterolemia and patients with significantly elevated LDL despite taking maximum doses of statins. Unlike other medications that target apoB-100, such as microsomal triglyceride transfer proteins, mipomersen does not cause hepatic steatosis or intestinal steatosis and does not affect dietary fat absorption. Adverse side effects encountered with mipomersen include flu-like symptoms, injection site reactions, and elevated liver transaminases. If future studies continue to show such promising results, mipomersen would likely be a viable additional lipid-lowering therapy for patients who are at high cardiovascular risk, intolerant to statins, and/or not at target lipid levels despite maximum doses of statin therapy. Clinical outcome studies looking at cardiovascular disease end points still need to be done.


Assuntos
Anticolesterolemiantes/uso terapêutico , Apolipoproteína B-100/antagonistas & inibidores , Doenças Cardiovasculares/prevenção & controle , Inibidores de Hidroximetilglutaril-CoA Redutases/uso terapêutico , Hipercolesterolemia/tratamento farmacológico , Oligonucleotídeos/uso terapêutico , Administração Cutânea , Anticolesterolemiantes/farmacocinética , Anticolesterolemiantes/farmacologia , Ensaios Clínicos como Assunto , Esquema de Medicação , Humanos , Oligonucleotídeos/farmacocinética , Oligonucleotídeos/farmacologia , Oligonucleotídeos Antissenso/farmacocinética , Oligonucleotídeos Antissenso/farmacologia , Oligonucleotídeos Antissenso/uso terapêutico
19.
Gynecol Endocrinol ; 27(10): 800-6, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21879795

RESUMO

The aim of the study was to verify whether post-menopausal hormone replacement therapy (HRT) modifies autoantibody titers against oxidized low-density lipoprotein (LDL) (anti-LDLoxi), against epitopes of oxidized apolipoprotein B100 and common carotid intima-media thickness (IMT) in these women. Sixty-eight women in pre-menopause (PMW) and 216 in post-menopause (POMW) were recruited; eighty-three had undergone HRT for at least 12 months, where 48 received conjugated estrogens alone (EHRT) and 35 received conjugated estrogen and medroxyprogesterone acetate (CHRT). ELISA was used to determine autoantibodies. Lipoprotein lipase (LPL), hepatic lipase (HL), cholesterol ester transfer protein (CETP) and phospholipid transfer protein (PLTP) activities were assayed by radiometric methods. IMT was measured using Doppler ultrasound. Anti-oxidized LDL and anti-D antibodies increased by 40% (p ≤ 0.003) and 42% (p ≤ 0.006), respectively, with menopause. There was a surprising and significant 7% reduction in anti-D2 antibody titers with HRT (p ≤ 0.050), indicating a positive effect of treatment on the immune response to oxidized LDL. Combined HRT decreased activities of HL and LPL. HRT did not change common carotid IMT, which was increased by 32% as expected after menopause (p ≤ 0.030). This study describes, for the first time, the protective effect of HRT on decreasing autoantibody titers against oxidized apolipoprotein B in LDL.


Assuntos
Apolipoproteína B-100/antagonistas & inibidores , Autoanticorpos/análise , Doenças Autoimunes/prevenção & controle , Terapia de Reposição de Estrogênios , Lipoproteínas LDL/antagonistas & inibidores , Menopausa/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Adulto , Idoso , Idoso de 80 Anos ou mais , Apolipoproteína B-100/química , Doenças Autoimunes/sangue , Doenças Autoimunes/imunologia , Doenças Autoimunes/patologia , Doenças Cardiovasculares/prevenção & controle , Artéria Carótida Primitiva/diagnóstico por imagem , Artéria Carótida Primitiva/efeitos dos fármacos , Artéria Carótida Primitiva/imunologia , Artéria Carótida Primitiva/patologia , Espessura Intima-Media Carotídea , Epitopos , Terapia de Reposição de Estrogênios/métodos , Estrogênios Conjugados (USP)/uso terapêutico , Feminino , Humanos , Lipase/sangue , Lipase Lipoproteica/sangue , Acetato de Medroxiprogesterona/uso terapêutico , Menopausa/imunologia , Pessoa de Meia-Idade , Oxirredução
20.
Am J Cardiovasc Drugs ; 10(4): 271-9, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20653334

RESUMO

Genzyme Corporation and Isis Pharmaceuticals have a worldwide licensing and collaboration agreement for the development of subcutaneous mipomersen (ISIS 147764; ISIS 301012; ISIS301012; mipomersen-sodium). Mipomersen, an oligonucleotide antisense inhibitor directed against apolipoprotein B-100 (apoB-100) mRNA, is in phase III clinical evaluation for the treatment of familial hypercholesterolemia, a form of type IIa hyperlipoproteinemia, and hypercholesterolemia in patients with severely high cholesterol levels or at high risk for coronary heart disease in the US, European Union, Brazil, Canada, South Africa, and South East Asia. This review discusses the development history and scientific profile of this new compound.


Assuntos
Apolipoproteína B-100/antagonistas & inibidores , Oligonucleotídeos Antissenso/farmacologia , Oligonucleotídeos/farmacologia , Animais , Ensaios Clínicos como Assunto , Humanos , Hipercolesterolemia/tratamento farmacológico , Hipercolesterolemia/fisiopatologia , Hiperlipoproteinemia Tipo II/tratamento farmacológico , Hiperlipoproteinemia Tipo II/fisiopatologia , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA