RESUMO
BACKGROUND: To date, no genome-wide association study (GWAS) has considered the combined phenotype of asthma with hay fever. Previous analyses of family data from the Tasmanian Longitudinal Health Study provide evidence that this phenotype has a stronger genetic cause than asthma without hay fever. OBJECTIVE: We sought to perform a GWAS of asthma with hay fever to identify variants associated with having both diseases. METHODS: We performed a meta-analysis of GWASs comparing persons with both physician-diagnosed asthma and hay fever (n = 6,685) with persons with neither disease (n = 14,091). RESULTS: At genome-wide significance, we identified 11 independent variants associated with the risk of having asthma with hay fever, including 2 associations reaching this level of significance with allergic disease for the first time: ZBTB10 (rs7009110; odds ratio [OR], 1.14; P = 4 × 10(-9)) and CLEC16A (rs62026376; OR, 1.17; P = 1 × 10(-8)). The rs62026376:C allele associated with increased asthma with hay fever risk has been found to be associated also with decreased expression of the nearby DEXI gene in monocytes. The 11 variants were associated with the risk of asthma and hay fever separately, but the estimated associations with the individual phenotypes were weaker than with the combined asthma with hay fever phenotype. A variant near LRRC32 was a stronger risk factor for hay fever than for asthma, whereas the reverse was observed for variants in/near GSDMA and TSLP. Single nucleotide polymorphisms with suggestive evidence for association with asthma with hay fever risk included rs41295115 near IL2RA (OR, 1.28; P = 5 × 10(-7)) and rs76043829 in TNS1 (OR, 1.23; P = 2 × 10(-6)). CONCLUSION: By focusing on the combined phenotype of asthma with hay fever, variants associated with the risk of allergic disease can be identified with greater efficiency.
Assuntos
Asma/diagnóstico , Asma/genética , Variação Genética , Estudo de Associação Genômica Ampla , Fenótipo , Locos de Características Quantitativas , Rinite Alérgica Sazonal/diagnóstico , Rinite Alérgica Sazonal/genética , Adulto , Alelos , Asma/complicações , Feminino , Frequência do Gene , Humanos , Lectinas Tipo C/genética , Desequilíbrio de Ligação , Masculino , Pessoa de Meia-Idade , Proteínas de Transporte de Monossacarídeos/genética , Razão de Chances , Polimorfismo de Nucleotídeo Único , Proteínas Repressoras/genética , Rinite Alérgica Sazonal/complicações , Adulto JovemRESUMO
The synthesis and antibacterial activity of heterocyclic methylsulfone hydroxamates is presented. Compounds in this series are potent inhibitors of the LpxC enzyme, a key enzyme involved in the production of lipopolysaccharide (LPS) found in the outer membrane of Gram-negative bacteria. SAR evaluation of compounds in this series revealed analogs with potent antibacterial activity against challenging Gram-negative species such as Pseudomonas aeruginosa and Klebsiella pneumoniae.
Assuntos
Amidoidrolases/antagonistas & inibidores , Antibacterianos/química , Inibidores Enzimáticos/química , Bactérias Gram-Negativas/efeitos dos fármacos , Ácidos Hidroxâmicos/química , Amidoidrolases/metabolismo , Antibacterianos/síntese química , Antibacterianos/farmacologia , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/farmacologia , Compostos Heterocíclicos/química , Ácidos Hidroxâmicos/síntese química , Ácidos Hidroxâmicos/farmacologia , Klebsiella pneumoniae/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/enzimologia , Relação Estrutura-Atividade , Sulfonas/químicaRESUMO
In recent years, the first generation of ß-secretase (BACE1) inhibitors advanced into clinical development for the treatment of Alzheimer's disease (AD). However, the alignment of drug-like properties and selectivity remains a major challenge. Herein, we describe the discovery of a novel class of potent, low clearance, CNS penetrant BACE1 inhibitors represented by thioamidine 5. Further profiling suggested that a high fraction of the metabolism (>95%) was due to CYP2D6, increasing the potential risk for victim-based drug-drug interactions (DDI) and variable exposure in the clinic due to the polymorphic nature of this enzyme. To guide future design, we solved crystal structures of CYP2D6 complexes with substrate 5 and its corresponding metabolic product pyrazole 6, which provided insight into the binding mode and movements between substrate/inhibitor complexes. Guided by the BACE1 and CYP2D6 crystal structures, we designed and synthesized analogues with reduced risk for DDI, central efficacy, and improved hERG therapeutic margins.
Assuntos
Secretases da Proteína Precursora do Amiloide/antagonistas & inibidores , Secretases da Proteína Precursora do Amiloide/química , Ácido Aspártico Endopeptidases/antagonistas & inibidores , Ácido Aspártico Endopeptidases/química , Citocromo P-450 CYP2D6/química , Interações Medicamentosas , Inibidores de Proteases/química , Inibidores de Proteases/farmacologia , Sequência de Aminoácidos , Proteínas Amiloidogênicas/metabolismo , Animais , Cristalografia por Raios X , Citocromo P-450 CYP2D6/genética , Citocromo P-450 CYP2D6/metabolismo , Desenho de Fármacos , Canal de Potássio ERG1 , Canais de Potássio Éter-A-Go-Go/antagonistas & inibidores , Canais de Potássio Éter-A-Go-Go/metabolismo , Humanos , Concentração Inibidora 50 , Masculino , Camundongos Endogâmicos , Modelos Moleculares , Dados de Sequência Molecular , Inibidores de Proteases/administração & dosagem , Inibidores de Proteases/farmacocinética , Pirazóis/química , Relação Estrutura-AtividadeRESUMO
The mechanism of necrotizing myopathy associated with antibodies to signal recognition particle (SRP) remains unclear. We investigated the effect of anti-SRP+serum and complement on cell viability in myoblast cultures. Cell viability was only slightly reduced by incubation with anti-SRP+serum compared with control serum. However, the addition of fresh complement resulted in a marked reduction in cell survival. Surface immunostaining for SRP, C3c and C5b-9 was demonstrated in cultures pre-incubated with anti-SRP+serum and complement, and in muscle biopsies from patients with myopathy. These findings provide further support for a complement-dependent antibody-mediated mechanism in anti-SRP associated myopathy.
Assuntos
Anticorpos/sangue , Complemento C3c/metabolismo , Doenças Musculares , Partícula de Reconhecimento de Sinal/imunologia , Idoso , Sobrevivência Celular , Células Cultivadas , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Células Musculares/metabolismo , Células Musculares/patologia , Músculo Esquelético/patologia , Doenças Musculares/imunologia , Doenças Musculares/metabolismo , Doenças Musculares/patologia , Mioblastos/metabolismo , Mioblastos/patologiaRESUMO
In the search for the most efficacious antisense oligonucleotides (AOs) aimed at inducing SMN2 exon 7 inclusion, we systematically assessed three AOs, PMO25 (-10, -34), PMO18 (-10, -27), and PMO20 (-10, -29), complementary to the SMN2 intron 7 splicing silencer (ISS-N1). PMO25 was the most efficacious in augmenting exon 7 inclusion in vitro in spinal muscular atrophy (SMA) patient fibroblasts and in vitro splicing assays. PMO25 and PMO18 were compared further in a mouse model of severe SMA. After a single intracerebroventricular (ICV) injection in neonatal mice, PMO25 increased the life span of severe SMA mice up to 30-fold, with average survival greater by 3-fold compared with PMO18 at a dose of 20 µg/g and 2-fold at 40 µg/g. Exon 7 inclusion was increased in the CNS but not in peripheral tissues. Systemic delivery of PMO25 at birth achieved a similar outcome and produced increased exon 7 inclusion both in the CNS and peripherally. Systemic administration of a 10-µg/g concentration of PMO25 conjugated to an octaguanidine dendrimer (VMO25) increased the life span only 2-fold in neonatal type I SMA mice, although it prevented tail necrosis in mild SMA mice. Higher doses and ICV injection of VMO25 were associated with toxicity. We conclude that (1) the 25-mer AO is more efficient than the 18-mer and 20-mer in modifying SMN2 splicing in vitro; (2) it is more efficient in prolonging survival in SMA mice; and (3) naked Morpholino oligomers are more efficient and safer than the Vivo-Morpholino and have potential for future SMA clinical applications.
Assuntos
Íntrons , Morfolinos/metabolismo , Atrofia Muscular Espinal/genética , Atrofia Muscular Espinal/metabolismo , Sequências Reguladoras de Ácido Nucleico , Proteína 2 de Sobrevivência do Neurônio Motor/genética , Proteína 2 de Sobrevivência do Neurônio Motor/metabolismo , Processamento Alternativo , Animais , Pareamento de Bases , Sequência de Bases , Modelos Animais de Doenças , Éxons , Ordem dos Genes , Humanos , Camundongos , Camundongos Transgênicos , Morfolinos/administração & dosagem , Morfolinos/química , Atrofia Muscular Espinal/mortalidade , Atrofia Muscular Espinal/terapiaRESUMO
Spinal muscular atrophy (SMA) is caused by loss of the Survival Motor Neuron 1 (SMN1) gene, resulting in reduced SMN protein. Humans possess the additional SMN2 gene (or genes) that does produce low level of full length SMN, but cannot adequately compensate for loss of SMN1 due to aberrant splicing. The majority of SMN2 gene transcripts lack exon 7 and the resultant SMNΔ7 mRNA is translated into an unstable and non-functional protein. Splice intervention therapies to promote exon 7 retention and increase amounts of full-length SMN2 transcript offer great potential as a treatment for SMA patients. Several splice silencing motifs in SMN2 have been identified as potential targets for antisense oligonucleotide mediated splice modification. A strong splice silencer is located downstream of exon 7 in SMN2 intron 7. Antisense oligonucleotides targeting this motif promoted SMN2 exon 7 retention in the mature SMN2 transcripts, with increased SMN expression detected in SMA fibroblasts. We report here systematic optimisation of phosphorodiamidate morpholino oligonucleotides (PMO) that promote exon 7 retention to levels that rescued the phenotype in a severe mouse model of SMA after intracerebroventricular delivery. Furthermore, the PMO gives the longest survival reported to date after a single dosing by ICV.
Assuntos
Morfolinos/genética , Atrofia Muscular Espinal/terapia , Oligonucleotídeos Antissenso/genética , Processamento Pós-Transcricional do RNA , Animais , Sequência de Bases , Células Cultivadas , Terapia Genética , Humanos , Injeções Intraventriculares , Camundongos , Camundongos Transgênicos , Atrofia Muscular Espinal/genética , Interferência de RNA , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteína 2 de Sobrevivência do Neurônio Motor/genética , Proteína 2 de Sobrevivência do Neurônio Motor/metabolismo , Titulometria , Transcrição GênicaRESUMO
Allergen-specific immunoglobulin E (present in allergic sensitization) has a central role in the pathogenesis of allergic disease. We performed the first large-scale genome-wide association study (GWAS) of allergic sensitization in 5,789 affected individuals and 10,056 controls and followed up the top SNP at each of 26 loci in 6,114 affected individuals and 9,920 controls. We increased the number of susceptibility loci with genome-wide significant association with allergic sensitization from three to ten, including SNPs in or near TLR6, C11orf30, STAT6, SLC25A46, HLA-DQB1, IL1RL1, LPP, MYC, IL2 and HLA-B. All the top SNPs were associated with allergic symptoms in an independent study. Risk-associated variants at these ten loci were estimated to account for at least 25% of allergic sensitization and allergic rhinitis. Understanding the molecular mechanisms underlying these associations may provide new insights into the etiology of allergic disease.
Assuntos
Loci Gênicos , Estudo de Associação Genômica Ampla , Hipersensibilidade/genética , Alelos , Biologia Computacional , Redes Reguladoras de Genes , Genômica , Humanos , Hipersensibilidade/metabolismo , Fenótipo , Polimorfismo de Nucleotídeo Único , Locos de Características Quantitativas , Transdução de SinaisRESUMO
ß-Lactamase inhibitors (clavulanic acid, sulbactam, and tazobactam) contribute significantly to the longevity of the ß-lactam antibiotics used to treat serious infections. In the quest to design more potent compounds and to understand the mechanism of action of known inhibitors, 6ß-(hydroxymethyl)penicillanic acid sulfone (6ß-HM-sulfone) was tested against isolates expressing the class A TEM-1 ß-lactamase and a clinically important variant of the AmpC cephalosporinase of Pseudomonas aeruginosa, PDC-3. The addition of the 6ß-HM-sulfone inhibitor to ampicillin was highly effective. 6ß-HM-sulfone inhibited TEM-1 with an IC(50) of 12 ± 2 nM and PDC-3 with an IC(50) of 180 ± 36 nM, and displayed lower partition ratios than commercial inhibitors, with partition ratios (k(cat)/k(inact)) equal to 174 for TEM-1 and 4 for PDC-3. Measured for 20 h, 6ß-HM-sulfone demonstrated rapid, first-order inactivation kinetics with the extent of inactivation being related to the concentration of inhibitor for both TEM-1 and PDC-3. Using mass spectrometry to gain insight into the intermediates of inactivation of this inhibitor, 6ß-HM-sulfone was found to form a major adduct of +247 ± 5 Da with TEM-1 and +245 ± 5 Da with PDC-3, suggesting that the covalently bound, hydrolytically stabilized acyl-enzyme has lost a molecule of water (HOH). Minor adducts of +88 ± 5 Da with TEM-1 and +85 ± 5 Da with PDC-3 revealed that fragmentation of the covalent adduct can result but appeared to occur slowly with both enzymes. 6ß-HM-sulfone is an effective and versatile ß-lactamase inhibitor of representative class A and C enzymes.
Assuntos
Antibacterianos/farmacologia , Proteínas de Bactérias/metabolismo , Sulbactam/análogos & derivados , Sulbactam/farmacologia , beta-Lactamases/metabolismo , Antibacterianos/química , Proteínas de Bactérias/antagonistas & inibidores , Proteínas de Bactérias/genética , Domínio Catalítico , Simulação por Computador , Escherichia coli/efeitos dos fármacos , Escherichia coli/enzimologia , Escherichia coli/genética , Modelos Moleculares , Estrutura Molecular , Conformação Proteica , Pseudomonas aeruginosa/enzimologia , Sulbactam/química , Inibidores de beta-Lactamases , beta-Lactamases/genéticaRESUMO
The synthesis and biological activity of a new series of LpxC inhibitors represented by pyridone methylsulfone hydroxamate 2a is presented. Members of this series have improved solubility and free fraction when compared to compounds in the previously described biphenyl methylsulfone hydroxamate series, and they maintain superior Gram-negative antibacterial activity to comparator agents.
Assuntos
Amidoidrolases/antagonistas & inibidores , Antibacterianos/síntese química , Bactérias Gram-Negativas/efeitos dos fármacos , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Ácidos Hidroxâmicos/síntese química , Piridonas/síntese química , Ácidos Sulfônicos/síntese química , Animais , Antibacterianos/farmacocinética , Antibacterianos/farmacologia , Cristalografia por Raios X , Humanos , Ácidos Hidroxâmicos/farmacocinética , Ácidos Hidroxâmicos/farmacologia , Testes de Sensibilidade Microbiana , Modelos Moleculares , Estrutura Molecular , Conformação Proteica , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/enzimologia , Piridonas/farmacocinética , Piridonas/farmacologia , Ratos , Estereoisomerismo , Relação Estrutura-Atividade , Ácidos Sulfônicos/farmacocinética , Ácidos Sulfônicos/farmacologiaRESUMO
In this paper, we present the synthesis and SAR as well as selectivity, pharmacokinetic, and infection model data for representative analogues of a novel series of potent antibacterial LpxC inhibitors represented by hydroxamic acid.
Assuntos
Amidoidrolases/antagonistas & inibidores , Antibacterianos/síntese química , Compostos de Bifenilo/síntese química , Ácidos Hidroxâmicos/síntese química , Éteres Fenílicos/síntese química , Infecções por Pseudomonas/tratamento farmacológico , Sulfetos/síntese química , Sulfonas/síntese química , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Compostos de Bifenilo/química , Compostos de Bifenilo/farmacologia , Domínio Catalítico , Cristalografia por Raios X , Farmacorresistência Bacteriana , Ligação de Hidrogênio , Ácidos Hidroxâmicos/química , Ácidos Hidroxâmicos/farmacologia , Camundongos , Modelos Moleculares , Conformação Molecular , Éteres Fenílicos/química , Éteres Fenílicos/farmacologia , Pseudomonas aeruginosa , Ratos , Estereoisomerismo , Relação Estrutura-Atividade , Sulfetos/química , Sulfetos/farmacologia , Sulfonas/química , Sulfonas/farmacologiaRESUMO
An efficient method was developed for the synthesis of 2-methylene-4-substituted ethyl butyrates via cyclopropyl opening followed by a Wittig reaction. The desired products were formed in a two-step, one-pot reaction sequence. Alternatively, the key intermediate ylide 2 was isolable and could be stored under oxygen-free conditions and subsequently utilized. A variety of nucleophiles were found to open the commercially available cyclopropane 1. The resulting ylide reacted with aldehydes to provide E-olefinic products.
Assuntos
Boratos/química , Compostos Organofosforados/química , Aldeídos/química , Ciclização , Estrutura MolecularRESUMO
As part of an effort to synthesize a dendronized cellulose, we have synthesized a trifunctional aminoamide derivative, which is the first generation of a dendron substituent. We anticipate that a dendronized cellulose would have applications in complexing metals and could be employed as an adjuvant for drugs. The trifunctional aminoamide substituent was introduced by coupling di-tert-butyl 4-[2-(tert-butoxycarbonyl)ethyl]-4-aminoheptanedicarboxylate, BA, directly to a (carboxymethyl)cellulose (CMC) backbone and converting the tert-butyl ester peripheral groups to aminoamide substituents by use of N,N-dimethyl-1,3-propanediamine. Confirmation of the proposed chemical structure of the intermediates as well as the water-soluble aminoamide derivative (CMCBADMPDA) was obtained by Fourier transform infrared (FT-IR) and NMR spectroscopy. The degree of substitution (DS) was determined to be 0.40 +/- 0.01 by thermogravimetric analysis. Typical weight average molecular weight (M(w)), molecular weight distribution (MWD), and molecular size of the dendronized polymers were found to be 97,000, 1.7, and 17.4 nm for derivatives of a CMC with corresponding M(w), MWD, and root-mean-square radius (RMS) of 230 000, 3.2, and 24 nm. A differential refractive index (dn/dc) for the aminoamide derivative measured in aqueous 0.40 N ammonium acetate-0.01 N NaOH was found to be 0.1473. The intrinsic viscosity of the dendronized cellulose decreased significantly when compared with that of CMC, that is, 0.40 dL/g relative to 5.60 dL/g. The hydrophobicity of the CMCBADMPDA microenvironment in aqueous solution was probed by evaluating the relative fluorescence intensities of the I(373)/I(384) pyrene bands; a slightly more hydrophobic environment was observed.