RESUMO
BACKGROUND: The trajectory from healthy to critical illness is influenced by numerous factors, including metabolism, which differs substantially between males and females. Whole body protein breakdown is substantially increased in critically ill patients, but it remains unclear whether there are sex differences that could explain the different health outcomes. Hence, we performed a secondary analysis of a study, where we used a novel pulse isotope method in critically ill and matched healthy males and females. METHODS: In 51 critically ill ICU patients (26 males, 15 females) and 49 healthy controls (36 males and 27 females), we assessed their general and disease characteristics and collected arterial(ized) blood in the postabsorptive state after pulse administration of 8 ml of a solution containing 18 stable AA tracers. In contrast to the original study, we now fitted the decay curves and calculated non-compartmental whole body amino acid production (WBP) and compartmental measurements of metabolism, including intracellular amino acid production. We measured amino acid enrichments and concentrations by LC-MS/MS and derived statistics using AN(C)OVA. RESULTS: Critically ill males and females showed an increase in the WBP of many amino acids, including those related to protein breakdown, but females showed greater elevations, or in the event of a reduction, attenuated reductions. Protein breakdown-independent WBP differences remained between males and females, notably increased glutamine and glutamate WBP. Only severely ill females showed a lower increase in WBP of many amino acids in comparison to moderately ill females, suggesting a suppressed metabolism. Compartmental analysis supported the observations. CONCLUSIONS: The present study shows that females have a different response to critical illness in the production of several amino acids and changes in protein breakdown, observations made possible using our innovative stable tracer pulse approach. CLINICAL TRIAL REGISTRY: Data are from the baseline measurements of study NCT02770092 (URL: https://clinicaltrials.gov/ct2/show/NCT02770092) and NCT03628365 (URL: https://clinicaltrials.gov/ct2/show/NCT03628365).
Assuntos
Aminoácidos , Estado Terminal , Feminino , Humanos , Masculino , Aminoácidos/metabolismo , Cromatografia Líquida , Espectrometria de Massas em TandemRESUMO
Amino acid (AA) metabolism is severely disturbed in critically ill ICU patients. To be able to make a more scientifically based decision on the type of protein or AA nutrition to deliver in ICU patients, comprehensive AA phenotyping with measurements of plasma concentrations and whole body production (WBP) is needed. Therefore, we studied ICU patients and matched control subjects using a novel pulse isotope method to obtain in-depth metabolic analysis. In 51 critically ill ICU patients (SOFA~6.6) and 49 healthy controls, we measured REE and body composition/phase-angle using BIA. In the postabsorptive state, we collected arterial (ized) blood for CRP and AA. Then, we administered an 8 mL solution containing 18 stable AA tracers as a pulse and calculated WBP. Enrichments: LC-MS/MS and statistics: t-test, ANCOVA. Compared to healthy, critically ill ICU patients had lower phase-angle (p < 0.00001), and higher CRP (p < 0.0001). Most AA concentrations were lower in ICU patients (p < 0.0001), except tau-methylhistidine and phenylalanine. WBP of most AA were significantly (p < 0.0001) higher with increases in glutamate (160%), glutamine (46%), and essential AA. Remarkably, net protein breakdown was lower. There were only weak relationships between AA concentrations and WBP. Critically ill ICU patients (SOFA 8-16) had lower values for phase angle (p = 0.0005) and small reductions of most plasma AA concentrations, but higher tau-methylhistidine (p = 0.0223) and hydroxyproline (p = 0.0028). Remarkably, the WBP of glutamate and glutamine were lower (p < 0.05), as was their clearance, but WBP of tau-methylhistidine (p = 0.0215) and hydroxyproline (p = 0.0028) were higher. Our study in critically ill ICU patients shows that comprehensive metabolic phenotyping was able to reveal severe disturbances in specific AA pathways, in a disease severity dependent way. This information may guide improving nutritional compositions to improve the health of the critically ill patient. CLINICAL TRIAL REGISTRY: Data are from the baseline measurements of study NCT02770092 (URL: https://clinicaltrials.gov/ct2/show/NCT02770092) and NCT03628365 (URL: https://clinicaltrials.gov/ct2/show/NCT03628365).
Assuntos
Aminoácidos/sangue , Composição Corporal/fisiologia , Idoso , Metabolismo Basal/fisiologia , Estado Terminal , Impedância Elétrica , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
The inbred mouse strain C57BL/6 is commonly used for the generation of transgenic mouse and is a well established strain in bone research. Different vendors supply different substrains of C57BL/6J as wild-type animals when genetic drift did not incur any noticeable phenotype. However, we sporadically observed drastic differences in the bone phenotype of "WT" C57BL/6J mice originating from different labs and speculated that these variations are attributable, at least in part, to the variation between C57BL/6J substrains, which is often overlooked. C57BL/6J-OlaHsd is a commonly used substrain that despite a well defined deletion in the alpha-synuclein (Snca) and multimerin-1 (Mmrn1) genes, was reported to display no obvious phenotype and is used as WT control. Here, we compared the bone phenotype of C57BL/6J-OlaHsd (6J-OLA) to C57BL/6J-RccHsd (6J-RCC) and to the original C57BL/6J (6J-JAX). Using µCT analysis, we found that 6J-OLA mice display a significantly lower trabecular bone mass compared to 6J-RCC and 6J-JAX. PCR analysis revealed that both the Snca and Mmrn1 genes are expressed in bone tissue of 6J-RCC animals but not of 6J-OLA mutants, suggesting either one or both genes play a role in bone metabolism. In vitro analysis demonstrated increase in osteoclasts number and decreased osteoblast mineralization in cells derived from 6J-OLA compared with 6J-RCC. Our data may shed light on unexplained differences in basal bone measurements between different research centers and reiterate the importance of specifying the exact substrain type. In addition, our findings describe the physiological role for Mmrn1 and/or Snca in bone remodeling.
Assuntos
Proteínas Sanguíneas/genética , Remodelação Óssea/genética , Moléculas de Adesão Celular/genética , Mutação , Osteoporose/genética , alfa-Sinucleína/genética , Animais , Proteínas Sanguíneas/metabolismo , Densidade Óssea , Calcificação Fisiológica , Moléculas de Adesão Celular/metabolismo , Células Cultivadas , Fêmur/diagnóstico por imagem , Fêmur/metabolismo , Fêmur/fisiopatologia , Predisposição Genética para Doença , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Osteoblastos/metabolismo , Osteoclastos/metabolismo , Osteogênese , Osteoporose/diagnóstico por imagem , Osteoporose/metabolismo , Osteoporose/fisiopatologia , Fenótipo , Microtomografia por Raio-X , alfa-Sinucleína/metabolismoRESUMO
Despite progress in systemic small interfering RNA (siRNA) delivery to the liver and to solid tumors, systemic siRNA delivery to leukocytes remains challenging. The ability to silence gene expression in leukocytes has great potential for identifying drug targets and for RNAi-based therapy for leukocyte diseases. However, both normal and malignant leukocytes are among the most difficult targets for siRNA delivery as they are resistant to conventional transfection reagents and are dispersed in the body. We used mantle cell lymphoma (MCL) as a prototypic blood cancer for validating a novel siRNA delivery strategy. MCL is an aggressive B-cell lymphoma that overexpresses cyclin D1 with relatively poor prognosis. Down-regulation of cyclin D1 using RNA interference (RNAi) is a potential therapeutic approach to this malignancy. Here, we designed lipid-based nanoparticles (LNPs) coated with anti-CD38 monoclonal antibodies that are specifically taken up by human MCL cells in the bone marrow of xenografted mice. When loaded with siRNAs against cyclin D1, CD38-targeted LNPs induced gene silencing in MCL cells and prolonged survival of tumor-bearing mice with no observed adverse effects. These results highlight the therapeutic potential of cyclin D1 therapy in MCL and present a novel RNAi delivery system that opens new therapeutic opportunities for treating MCL and other B-cell malignancies.
Assuntos
Linfócitos B/imunologia , Linfoma de Células B/terapia , Linfoma de Célula do Manto/terapia , Nanomedicina/métodos , Interferência de RNA , RNA Interferente Pequeno/administração & dosagem , ADP-Ribosil Ciclase 1/imunologia , Animais , Anticorpos Monoclonais/imunologia , Linhagem Celular Tumoral , Ciclina D1/genética , Regulação para Baixo , Inativação Gênica , Humanos , Lipídeos , Linfoma de Células B/imunologia , Linfoma de Célula do Manto/genética , Linfoma de Célula do Manto/imunologia , Camundongos , Nanopartículas , RNA Interferente Pequeno/genética , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
BACKGROUND: Nuclear factor (NF)-κB is a transcription factor that regulates cytokine and chemokine production in various inflammatory diseases, including bronchial asthma. IκB kinase (IKK) ß is important for NF-κB activation in inflammatory conditions, and is possibly related to airway remodelling. Thus, inhibition of the IKKß-NF-κB pathway may be an ideal strategy for the management of airway remodelling. OBJECTIVE: We examined the effects of a newly synthesized IKKß inhibitor, IMD-0354, in a chronic allergen exposure model of bronchial asthma in mice. METHODS: A chronic mouse model was generated by challenge with house dust mite antigen (Dermatophagoides pteronyssinus). IMD-0354 was administrated intraperitoneally in therapeutic groups. Lung histopathology, hyperresponsiveness and the concentrations of mediators and molecules in supernatants of lung homogenates were determined. RESULTS: NF-κB activation was inhibited by prolonged periods of IMD-0354 administration. IMD-0354 reduced the numbers of bronchial eosinophils. IMD-0354 also inhibited the pathological features of airway remodelling, including goblet cell hyperplasia, subepithelial fibrosis, collagen deposition and smooth muscle hypertrophy. Inhibition of these structural changes by IMD-0354 was the result of the suppressing the production and activation of remodelling-related mediators, such as TGF-ß, via inhibition of IKKß. IMD-0354 inhibited IL-13 and IL-1ß production, and it restored the production of IFN-γ. It also ameliorated airway hyperresponsiveness. CONCLUSION: IKKß plays crucial roles in airway inflammation and remodelling in a chronic mouse model of asthma. A specific IKKß inhibitor, IMD-0354, may be therapeutically beneficial for treating airway inflammation and remodelling in chronic asthma.
Assuntos
Remodelação das Vias Aéreas/efeitos dos fármacos , Antígenos de Dermatophagoides/imunologia , Asma/tratamento farmacológico , Asma/patologia , Benzamidas/farmacologia , Modelos Animais de Doenças , Quinase I-kappa B/antagonistas & inibidores , Remodelação das Vias Aéreas/imunologia , Animais , Asma/enzimologia , Asma/fisiopatologia , Benzamidas/química , Benzamidas/uso terapêutico , Doença Crônica , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Estrutura MolecularRESUMO
Although it has been repeatedly suggested that low bone mineral density (BMD) is disproportionately prevalent among patients with depressive disorders, so far depression has not been officially acknowledged as a risk factor for osteoporosis. In a recent meta-analysis comparing depressed with nondepressed individuals we report that BMD is lower in depressed than nondepressed subjects. The association between depression and BMD is stronger in women than men, and in premenopausal than postmenopausal women. Only women psychiatrically diagnosed for major depression display significantly low BMD; women diagnosed by self-rating questionnaires do not. Using a mouse model for depression, we demonstrate a causal relationship between depressive-like behavior and bone loss. The depression-induced bone loss is associated with increases in skeletal norepinephrine and serum corticosterone levels. Bone loss, but not the depressive behavior, could be prevented by a beta-blocker. Hence, depression appears as a significant risk factor for low BMD, causing bone loss through stimulation of the sympathetic nervous system.
Assuntos
Doenças Ósseas Metabólicas/etiologia , Osso e Ossos/anatomia & histologia , Depressão/complicações , Depressão/fisiopatologia , Animais , Densidade Óssea/fisiologia , Doenças Ósseas Metabólicas/complicações , Depressão/metabolismo , Feminino , Humanos , Masculino , Camundongos , Fatores de RiscoRESUMO
Lymphoid neoplasms including lymphoma and leukemia are one of the most life-threatening disorders in dogs. Many lymphoid malignancies are well-treated with glucocorticoid (GC); however, GC resistance sometimes develops and its mechanism remains uncertain. Since constitutive activation of nuclear factor-κB (NF-κB) has been reported to play roles in lymphoid malignancies, we examined whether inhibition of NF-κB activity with a synthetic inhibitor IMD-0354 affected GC sensitivity of canine neoplastic lymphoid cells, CL-1 and GL-1. Dexamethasone failed to inhibit proliferation of these cells, in which low expression of glucocorticoid receptors (GR) was identified. In the presence of IMD-0354, GR expressions in CL-1 and GL-1 were increased, consequently dexamethasone inhibited their proliferation. These results indicated that GR expression might be down-regulated by spontaneous activation of NF-κB, resulting in GC resistance. Taken together, interference of NF-κB activity may have the synergistic effect in combination chemotherapy with GC for treatment against lymphoid malignancies.
Assuntos
Benzamidas/farmacologia , Doenças do Cão/tratamento farmacológico , Linfócitos/efeitos dos fármacos , Linfoma de Células B/veterinária , Linfoma de Células T/veterinária , NF-kappa B/antagonistas & inibidores , Receptores de Glucocorticoides/efeitos dos fármacos , Animais , Western Blotting/veterinária , Dexametasona/farmacologia , Doenças do Cão/imunologia , Cães , Relação Dose-Resposta a Droga , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/genética , Linfócitos/metabolismo , Linfoma de Células B/tratamento farmacológico , Linfoma de Células B/metabolismo , Linfoma de Células T/tratamento farmacológico , Linfoma de Células T/metabolismo , NF-kappa B/biossíntese , NF-kappa B/genética , Receptores de Glucocorticoides/biossíntese , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Regulação para Cima/efeitos dos fármacosRESUMO
Persimmon (Diospyros kaki Thunb.) is an important fruit in Asian countries, where it is eaten as a fresh fruit and is also used for many other purposes. To understand the molecular mechanism of fruit development and ripening in persimmon, we generated a total of 9,952 expressed sequence tags (ESTs) from randomly selected clones of two different cDNA libraries. One cDNA library was derived from fruit of "Saijo" persimmon at an early stage of development, and the other from ripening fruit. These ESTs were clustered into 6,700 non-redundant sequences. Of the 6,700 non-redundant sequences evaluated, the deduced amino acid sequences of 4,356 (65%) showed significant homology to known proteins, and 2,344 (35%) showed no significant similarity to any known proteins in Arabidopsis databases. We report comparison of genes identified in the two cDNA libraries and describe some putative genes involved in proanthocyanidin and carotenoid synthesis. This study provides the first global overview of a set of genes that are expressed during fruit development and ripening in persimmon.
Assuntos
Diospyros/genética , Etiquetas de Sequências Expressas , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Diospyros/crescimento & desenvolvimento , Frutas/genética , Frutas/crescimento & desenvolvimento , Biblioteca Gênica , Modelos Biológicos , Reação em Cadeia da Polimerase , Análise de Sequência de DNARESUMO
Since the discovery of the endocannabinoid system, its presence and involvement have been reported in a handful of biological systems. Recently, the skeleton has been identified as a major endocannabinoid target through both the neuronal CB1 and predominantly peripheral CB2 cannabinoid receptors. CB1 is present in sympathetic nerve terminals in bone, whereas CB2 is expressed in osteoblasts and osteoclasts, the respective bone-forming and -resorbing cells. Furthermore, the skeleton appears as the main system physiologically regulated by CB2. CB2-deficient mice show a markedly accelerated age-related bone loss and the CB2 locus in women is associated with low bone density and osteoporotic fractures. Since activation of CB2 attenuates experimentally induced bone loss by inhibiting bone resorption and stimulating bone formation, and because synthetic cannabinoids are stable and orally available, a therapy based on synthetic CB2 agonists is a promising novel target for antiosteoporotic drug development.
Assuntos
Remodelação Óssea , Moduladores de Receptores de Canabinoides/fisiologia , Endocanabinoides , Diferenciação Celular , Humanos , LigantesRESUMO
In Japanese pear (Pyrus pyrifolia Nakai), fruit storage potential is closely related to the amount of ethylene produced. We have developed a rapid and accurate method for analyzing genes involved in high ethylene production during fruit ripening in Japanese pear. This involves cleaved-amplified polymorphic sequences (CAPS) of two 1-aminocyclopropane-1-carboxylate (ACC) synthase genes (PPACS1 and PPACS2). Two CAPS markers (A for PPACS1 and B for PPACS2), associated with the amount of ethylene produced, were identified. Marker A was associated with high ethylene producers and marker B with moderate ethylene producers. The absence of these two markers enabled the identification of low ethylene producers. Using these markers, we have identified ethylene genotypes for 40 Japanese pear cultivars and two Chinese pear (P. bretschneideri) cultivars that are commercially important and used in breeding programs. Furthermore, we performed linkage analysis of these two genes in the F(2) population, which revealed that the recombination frequency between the two markers was 20.8 +/- 3.6%. This information is critical to the selection of parents and in breeding strategies to improve storage ability of Japanese pears.
Assuntos
Marcadores Genéticos , Liases/genética , Pyrus/enzimologia , Alelos , Sequência de Bases , Clonagem Molecular , Primers do DNA , Genótipo , Polimorfismo GenéticoRESUMO
PPARgamma is a ligand-activated transcription factor and functions as a heterodimer with a retinoid X receptor (RXR). Supraphysiological activation of PPARgamma by thiazolidinediones can reduce insulin resistance and hyperglycemia in type 2 diabetes, but these drugs can also cause weight gain. Quite unexpectedly, a moderate reduction of PPARgamma activity observed in heterozygous PPARgamma-deficient mice or the Pro12Ala polymorphism in human PPARgamma, has been shown to prevent insulin resistance and obesity induced by a high-fat diet. In this study, we investigated whether functional antagonism toward PPARgamma/RXR could be used to treat obesity and type 2 diabetes. We show herein that an RXR antagonist and a PPARgamma antagonist decrease triglyceride (TG) content in white adipose tissue, skeletal muscle, and liver. These inhibitors potentiated leptin's effects and increased fatty acid combustion and energy dissipation, thereby ameliorating HF diet-induced obesity and insulin resistance. Paradoxically, treatment of heterozygous PPARgamma-deficient mice with an RXR antagonist or a PPARgamma antagonist depletes white adipose tissue and markedly decreases leptin levels and energy dissipation, which increases TG content in skeletal muscle and the liver, thereby leading to the re-emergence of insulin resistance. Our data suggested that appropriate functional antagonism of PPARgamma/RXR may be a logical approach to protection against obesity and related diseases such as type 2 diabetes.
Assuntos
Diabetes Mellitus Tipo 2/metabolismo , Obesidade/metabolismo , Receptores Citoplasmáticos e Nucleares/antagonistas & inibidores , Receptores do Ácido Retinoico/antagonistas & inibidores , Tiazolidinedionas , Fatores de Transcrição/antagonistas & inibidores , Células 3T3 , Tecido Adiposo/metabolismo , Animais , Compostos Benzidrílicos , Benzoatos/metabolismo , Benzoatos/farmacologia , Compostos de Bifenilo/metabolismo , Compostos de Bifenilo/farmacologia , Compostos de Epóxi/metabolismo , Compostos de Epóxi/farmacologia , Ácidos Graxos/metabolismo , Hiperglicemia/etiologia , Hiperglicemia/metabolismo , Hipoglicemiantes/metabolismo , Hipoglicemiantes/farmacologia , Resistência à Insulina , Leptina/metabolismo , Camundongos , Camundongos Knockout , Ácidos Nicotínicos/metabolismo , Ácidos Nicotínicos/farmacologia , Receptores Adrenérgicos beta 3/metabolismo , Receptores Citoplasmáticos e Nucleares/agonistas , Receptores Citoplasmáticos e Nucleares/metabolismo , Receptores do Ácido Retinoico/agonistas , Receptores do Ácido Retinoico/metabolismo , Receptores X de Retinoides , Rosiglitazona , Tetra-Hidronaftalenos/metabolismo , Tetra-Hidronaftalenos/farmacologia , Tiazóis/metabolismo , Tiazóis/farmacologia , Fatores de Transcrição/agonistas , Fatores de Transcrição/metabolismoRESUMO
We have investigated the structure activity relationships of the potent retinoid agonist, 4-[4-(2-propyl-1,2-dicarba-closo-dodecaboran-l-yl)phenylamino]benzoic acid (BR403), which we have previously reported. Substitution of a methyl group on the aromatic nucleus or a methyl group on the nitrogen atom, or replacement of the amino group with ether, methylene, carboxyl or 1,1-ethylene greatly decreased the activity. The relatively planar conformation at the phenyl-N-phenyl moiety seems to play a critical role in the appearance of the biological activity.
Assuntos
Boranos/síntese química , Retinoides/agonistas , Animais , Benzoatos/química , Benzoatos/farmacologia , Boranos/química , Boranos/farmacologia , Terapia por Captura de Nêutron de Boro , Células COS , Diferenciação Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Células HL-60 , Humanos , Modelos Moleculares , Conformação Molecular , Receptores do Ácido Retinoico/agonistas , Relação Estrutura-Atividade , Ativação Transcricional/efeitos dos fármacosRESUMO
Aldose reductase (AR) has been implicated in the etiology of diabetic complications. Due to the limited number of currently available drugs for the treatment of diabetic complications, we have carried out structure-based drug design and synthesis in an attempt to find new types of AR inhibitors. With the ADAM&EVE program, a three-dimensional database (ACD3D) was searched using the ligand binding site of the AR crystal structure. Out of 179 compounds selected through this search followed by visual inspection, 36 compounds were purchased and subjected to a biological assay. Ten compounds showed more than 40% inhibition of AR at a 15 microg/mL concentration. In a subsequent lead optimization, a series of analogues of the most active compound were synthesized based on the docking mode derived by ADAM&EVE. Many of these congeners exhibited higher activities compared to the mother compound. Indeed, the most potent, synthesized compound showed an approximately 20-fold increase in inhibitory activity (IC(50) = 0.21 vs 4.3 microM). Furthermore, a hydrophobic subsite was newly inferred, which would be useful for the design of inhibitors with improved affinity for AR.
Assuntos
Aldeído Redutase/antagonistas & inibidores , Aldeído Redutase/química , Inibidores Enzimáticos/química , Indóis/química , Sítios de Ligação , Técnicas de Química Combinatória , Cristalografia por Raios X , Bases de Dados Factuais , Desenho de Fármacos , Inibidores Enzimáticos/síntese química , Indóis/síntese química , Ligantes , Modelos Moleculares , EstereoisomerismoRESUMO
BACKGROUND: Carboranes (dicarba-closo-dodecaboranes) are a class of carbon-containing polyhedral boron-cluster compounds having remarkable thermal stability and exceptional hydrophobicity. Applications of the unique structural and chemical properties offered by icosahedral carboranes in boron neutron capture therapy have received increasing attention over the past 30 years. However, these features of carboranes may allow another application as a hydrophobic pharmacophore in biologically active molecules that interact hydrophobically with receptors. RESULTS: We have designed candidate estrogen-receptor-binding compounds having carborane as a hydrophobic skeletal structure and synthesized them. The most potent compound bearing a carborane cage exhibited activity at least 10-fold greater than that of 17beta-estradiol in the luciferase reporter gene assay. Estrogen receptor-alpha-binding data for the compound were consistent with the results of the luciferase reporter gene assay. The compound also showed potent in vivo effects on the recovery of uterine weight and bone loss in ovariectomized mice. CONCLUSION: Further development of the potent carborane-containing estrogenic agonists described here, having a new skeletal structure and unique characteristics, should yield novel therapeutic agents, especially selective estrogen receptor modulators. Furthermore, the suitability of the spherical carborane cage for binding to the cavity of the estrogen receptor-alpha ligand-binding domain should provide a basis for a similar approach to developing novel ligands for other steroid receptors.
Assuntos
Compostos de Boro/síntese química , Compostos de Boro/uso terapêutico , Desenho de Fármacos , Estrogênios/síntese química , Estrogênios/uso terapêutico , Receptores de Estrogênio/agonistas , Animais , Densidade Óssea/efeitos dos fármacos , Compostos de Boro/metabolismo , Compostos de Boro/farmacologia , Células COS , Estradiol/química , Estradiol/metabolismo , Estradiol/farmacologia , Estrogênios/metabolismo , Estrogênios/farmacologia , Feminino , Fêmur/efeitos dos fármacos , Fêmur/patologia , Genes Reporter/genética , Ligação de Hidrogênio , Ligantes , Camundongos , Modelos Moleculares , Tamanho do Órgão/efeitos dos fármacos , Osteoporose/tratamento farmacológico , Osteoporose/patologia , Ratos , Receptores de Estrogênio/metabolismo , Relação Estrutura-Atividade , Ativação Transcricional/efeitos dos fármacos , Transfecção , Doenças Uterinas/tratamento farmacológico , Doenças Uterinas/patologia , Útero/efeitos dos fármacos , Útero/patologiaRESUMO
Aldose reductase has been implicated in the etiology of diabetic complications. Due to the limited number of currently available drugs for the treatment of diabetic complications, we have carried out a structure-based de novo design and synthesis in an attempt to identify new aldose reductase inhibitors. With the LEGEND program, we have designed 200 chemical structures that fit into the ligand binding site of the crystal structure of the enzyme. After their visual inspection and assessment of synthetic feasibility, four compounds were chosen to be synthesized. These compounds were all found to have reasonable inhibitory activities (IC50 = 17-91 microM), indicating the first successful generation of nonpeptide drug leads that have been obtained using a rational de novo design approach.
Assuntos
Aldeído Redutase/antagonistas & inibidores , Inibidores Enzimáticos/síntese química , Aldeído Redutase/química , Técnicas de Química Combinatória/métodos , Inibidores Enzimáticos/química , Modelos Moleculares , Conformação ProteicaAssuntos
Desenho de Fármacos , Genoma Humano , Genômica , Farmacogenética , Animais , Humanos , Tecnologia FarmacêuticaRESUMO
The discovery of a series of nonpeptide arginine vasopressin V(2) receptor agonists is described. After identifying the aniline derivative 8 as our lead compound from the metabolites of compound 7 that showed antidiuretic activity by po administration to Brattleboro rats, improvements in the in vitro potency involving evaluations of the structural requirements for agonist action and optimizing the structure of the benzoyl moiety have been intensively undertaken. These studies led to compounds 16g, 19a, and 23b,h,i that show potent agonist activity for the V(2) receptor.
Assuntos
Arginina Vasopressina/metabolismo , Benzazepinas/síntese química , Receptores de Vasopressinas/agonistas , Administração Oral , Animais , Benzazepinas/química , Benzazepinas/farmacologia , AMP Cíclico/biossíntese , Diurese/efeitos dos fármacos , Desenho de Fármacos , Feminino , Células HeLa , Humanos , Modelos Moleculares , Conformação Molecular , Ensaio Radioligante , Ratos , Receptores de Vasopressinas/metabolismo , Relação Estrutura-AtividadeRESUMO
Several pyridine- and pyrimidine-carboxylic acids were synthesized as ligand candidates for retinoid nuclear receptors, retinoic acid receptors (RARs) and retinoic X receptors (RXRs). Although the pyridine derivatives, 6-[(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthalenyl)carbamoyl]pyri dine-3-carboxylic acid (2b) and 6-[(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthalenyl)carboxamido]py ridine-3-carboxylic acid (5b) are more potent than the corresponding benzoic acid-type retinoids, Am80 (2a) and Am580 (5a), the replacement of the benzene ring of Am580 (5a), Am555 (6a), or Am55 (7a) with a pyrimidine ring caused loss of the retinoidal activity both in HL-60 cell differentiation assay and in RAR transactivation assay using COS-1 cells. On the other hand, pyrimidine analogs (PA series, 10 and 11) of potent RXR agonists (retinoid synergists) with a diphenylamine skeleton (DA series, 8 and 9) exhibited potent retinoid synergistic activity in HL-60 cell differentiation assay and activated RXRs. Among the synthesized compounds, 2-[N-n-propyl-N-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthalenyl)a mino]pyrimidine-5-carboxylic acid (PA013, 10e) is most active retinoid synergist in HL-60 assay.
Assuntos
Ácidos Carboxílicos/química , Pirimidinas/química , Retinoides/química , Animais , Células COS , Ácidos Carboxílicos/farmacologia , Diferenciação Celular/efeitos dos fármacos , Células HL-60 , Humanos , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Retinoides/farmacologia , Ativação Transcricional/efeitos dos fármacosRESUMO
Thirty cDNA clones of genes corresponding to mRNAs up-regulated during fruit ripening of Japanese pear (Pyrus pyrifolia Nakai cv. Kikusui) were obtained by differential screening of a ripe fruit cDNA library. All of these cDNAs were sequenced and gathered into 11 non-redundant groups after database searches. These clones represented genes associated with stress responses, protein catabolism or pathogenesis. The accumulation of transcripts of 3 out of 11 genes was inhibited by 1-methylcyclopropene (MCP), an inhibitor of ethylene action.
Assuntos
Etilenos/metabolismo , Frutas/genética , Rosales/genética , Transdução de Sinais , Northern Blotting , DNA Complementar/isolamento & purificação , Frutas/metabolismo , Frutas/fisiologia , RNA Mensageiro/análise , RNA de Plantas/análise , Rosales/metabolismo , Rosales/fisiologiaRESUMO
Dicarba-closo-dodecaboranes (carboranes), which have spherical geometry and hydrophobicity, are applicable as a hydrophobic pharmacophore of biologically active molecules. We have investigated structure-activity relations based on the structure of the potent estrogenic agonist, 1-hydroxymethyl-12-(4-hydroxyphenyl)-1,12-dicarba-closo-d odecaborane, which we have previously reported. The geometry and separation distance of the phenolic and alcoholic hydroxyl groups play a critical role in the appearance of biological activity.