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1.
J Cell Mol Med ; 22(5): 2865-2883, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29504694

RESUMO

Metastasis is the primary cause of death in prostate cancer (PCa) patients. Effective therapeutic intervention in metastatic PCa is undermined by our poor understanding of its molecular aetiology. Defining the mechanisms underlying PCa metastasis may lead to insights into how to decrease morbidity and mortality in this disease. Glyoxalase 1 (Glo1) is the detoxification enzyme of methylglyoxal (MG), a potent precursor of advanced glycation end products (AGEs). Hydroimidazolone (MG-H1) and argpyrimidine (AP) are AGEs originating from MG-mediated post-translational modification of proteins at arginine residues. AP is involved in the control of epithelial to mesenchymal transition (EMT), a crucial determinant of cancer metastasis and invasion, whose regulation mechanisms in malignant cells are still emerging. Here, we uncover a novel mechanism linking Glo1 to the maintenance of the metastatic phenotype of PCa cells by controlling EMT by engaging the tumour suppressor miR-101, MG-H1-AP and TGF-ß1/Smad signalling. Moreover, circulating levels of Glo1, miR-101, MG-H1-AP and TGF-ß1 in patients with metastatic compared with non-metastatic PCa support our in vitro results, demonstrating their clinical relevance. We suggest that Glo1, together with miR-101, might be potential therapeutic targets for metastatic PCa, possibly by metformin administration.


Assuntos
Transição Epitelial-Mesenquimal , Lactoilglutationa Liase/metabolismo , Neoplasias da Próstata/enzimologia , Neoplasias da Próstata/patologia , Regiões 3' não Traduzidas/genética , Idoso , Sequência de Bases , Linhagem Celular Tumoral , Movimento Celular , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Homoarginina/análogos & derivados , Homoarginina/sangue , Homoarginina/metabolismo , Humanos , Imidazóis/sangue , Imidazóis/metabolismo , Lactoilglutationa Liase/sangue , Masculino , Metformina/farmacologia , MicroRNAs/sangue , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Modelos Biológicos , Invasividade Neoplásica , Metástase Neoplásica , Ornitina/análogos & derivados , Ornitina/sangue , Ornitina/metabolismo , Fenótipo , Neoplasias da Próstata/sangue , Neoplasias da Próstata/genética , Pirimidinas/sangue , Pirimidinas/metabolismo , Transdução de Sinais , Proteínas Smad/metabolismo , Tioléster Hidrolases/metabolismo , Fator de Crescimento Transformador beta1/sangue , Fator de Crescimento Transformador beta1/metabolismo
2.
Amino Acids ; 47(9): 1697-702, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26210755

RESUMO

The year 2008 witnessed the first report on the increase in the concentration of L-homoarginine (hArg) in the maternal plasma during human pregnancy. This observation, along with a well-known function of hArg, the methylene homologue of L-arginine (Arg), as a substrate for nitric oxide (NO) synthase, was the ignition for the start of intense research on the physiology and pathology of hArg. The circulating concentration of hArg was found to be lower in patients suffering from various diseases, and hArg emerged within only very few years as a novel cardiovascular risk factor. The compendium in hand comprises original and review articles covering several aspects of hArg, Arg and its symmetrically and asymmetrically guanidine (N (G))-dimethylated derivatives SDMA and ADMA, respectively. In contrast to ADMA and SDMA, low hArg concentrations in plasma or serum and in urine are associated with high risks for morbidity and mortality, notably in the renal and cardiovascular systems. Acutely and chronically administered Arg as a nutritional supplement or in the form of dietary proteins is safe in animals and humans and leads to concomitant formation of hArg and ADMA, albeit in a different hArg/ADMA ratio. Despite the close but opposite associations of hArg and ADMA with disease in adults, children and adolescents, the underlying biochemical processes are largely unknown, presumably not restricted to NO, and warrant deeper investigation. As the common substrate for hArg and ADMA, Arg may play a key role in the biosynthesis and homeostasis of hArg and ADMA, two putative antagonists. In animal models of stroke and obesity, hArg has beneficial effects. The potential utility of hArg as a therapeutic drug or nutritional supplement in humans and animals remains to be elaborated.


Assuntos
Homoarginina , Obesidade , Acidente Vascular Cerebral , Adolescente , Adulto , Animais , Transporte Biológico Ativo , Modelos Animais de Doenças , Feminino , Homoarginina/análogos & derivados , Homoarginina/sangue , Homoarginina/urina , Humanos , Masculino , Obesidade/sangue , Obesidade/urina , Gravidez , Acidente Vascular Cerebral/sangue , Acidente Vascular Cerebral/urina
4.
J Biochem ; 144(1): 21-32, 2008 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-18344542

RESUMO

alphaA-crystallin is abundant in the lens of the eye and acts as a molecular chaperone by preventing aggregation of denaturing proteins. We previously found that chemical modification of the guanidino group of selected arginine residues by a metabolic alpha-dicarbonyl compound, methylglyoxal (MGO), makes human alphaA-crystallin a better chaperone. Here, we examined how the introduction of additional guanidino groups and modification by MGO influence the structure and chaperone function of alphaA-crystallin. alphaA-crystallin lysine residues were converted to homoarginine by guanidination with o-methylisourea (OMIU) and then modified with MGO. LC-ESI-mass spectrometry identified homoargpyrimidine and homohydroimidazolone adducts after OMIU and MGO treatment. Treatment with 0.25 M OMIU abolished most of the chaperone function. However, subsequent treatment with 1.0 mM MGO not only restored the chaperone function but increased it by approximately 40% and approximately 60% beyond that of unmodified alphaA-crystallin, as measured with citrate synthase and insulin aggregation assays, respectively. OMIU treatment reduced the surface hydrophobicity but after MGO treatment, it was approximately 39% higher than control. FRET analysis revealed that alphaA-crystallin subunit exchange rate was markedly retarded by OMIU modification, but was enhanced after MGO modification. These results indicate a pattern of loss and gain of chaperone function within the same protein that is associated with introduction of guanidino groups and their neutralization. These findings support our hypothesis that positively charged guanidino group on arginine residues keeps the chaperone function of alphaA-crystallin in check and that a metabolic alpha-dicarbonyl compound neutralizes this charge to restore and enhance chaperone function.


Assuntos
Cristalinas/química , Chaperonas Moleculares/química , Aminoácidos/análise , Cromatografia Líquida de Alta Pressão , Cristalinas/metabolismo , Homoarginina/análogos & derivados , Homoarginina/análise , Homoarginina/química , Humanos , Interações Hidrofóbicas e Hidrofílicas , Imidazóis/análise , Imidazóis/química , Espectrometria de Massas , Compostos de Metilureia/química , Chaperonas Moleculares/metabolismo , Ornitina/análogos & derivados , Ornitina/análise , Conformação Proteica , Subunidades Proteicas/química , Pirimidinas/análise , Pirimidinas/química , Aldeído Pirúvico/química
6.
Arch Pharm Res ; 27(1): 83-5, 2004 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-14969344

RESUMO

A sesquiterpene lactone, 1-O-acetyl-4R,6S-britannilactone (1) isolated from the flowers of Inula britannica L. var. chinensis (Rupr.) Reg. (Compositae), was found as an iNOS inhibitory constituent for the first time with an IC50 value of 22.1 microM which is more potent than the positive control, L-N6-(1-iminoethyl)lysine (IC50 = 33.7 microM). Structure of compound 1 was identified by 1D and 2D NMR experiments and by comparison with the reference standard.


Assuntos
Inula , Lipopolissacarídeos/química , Lipopolissacarídeos/farmacologia , Óxido Nítrico Sintase/química , Óxido Nítrico/antagonistas & inibidores , Extratos Vegetais/química , Avaliação Pré-Clínica de Medicamentos/métodos , Flores/química , Homoarginina/análogos & derivados , Homoarginina/farmacologia , Lactonas/química , Lactonas/isolamento & purificação , Lactonas/farmacologia , Lipopolissacarídeos/antagonistas & inibidores , Medicina Tradicional do Leste Asiático , Metanol , Estrutura Molecular , Óxido Nítrico Sintase/isolamento & purificação , Óxido Nítrico Sintase/farmacologia , Óxido Nítrico Sintase Tipo II , Sesquiterpenos/química , Sesquiterpenos/isolamento & purificação , Sesquiterpenos/farmacologia
7.
FASEB J ; 17(10): 1298-300, 2003 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-12738811

RESUMO

The inducible isoenzyme of nitric oxide synthase (iNOS) generates nitric oxide (NO) in inflammatory diseases such as asthma. The prodrug L-N6-(1-iminoethyl)lysine 5-tetrazole amide (SC-51) is rapidly converted in vivo to the active metabolite L-N6-(1-iminoethyl)lysine (L-NIL). Initially, we performed in vitro experiments in human primary airway epithelial cells to demonstrate that L-NIL causes inhibition of iNOS. In a randomized double-blind placebo-controlled crossover trial, SC-51 was administered as a single oral dose (20 or 200 mg) in separate cohorts of healthy volunteers (two groups of n=12) and mild asthmatic patients (two groups of n=12). SC-51 (200 mg) reduced exhaled breath NO levels to <2 ppb in both healthy volunteers (P<0.001) and mild asthmatics (P<0.001) within 15 min, representing >90% inhibition of baseline levels of NO in asthmatic patients, with the effects lasting at least 72 h. There were no significant effects on blood pressure, pulse rate, or respiratory function (FEV1). This study demonstrates that an inhibitor of iNOS produces marked inhibition of exhaled breath NO in normal and asthmatic subjects without producing the side effects observed following the systemic administration of non-selective NOS inhibitors, and thus provides support for the potential use of iNOS inhibitors to treat a range of inflammatory clinical disorders.


Assuntos
Asma/tratamento farmacológico , Inibidores Enzimáticos/farmacologia , Homoarginina/análogos & derivados , Homoarginina/farmacologia , Lisina/análogos & derivados , Óxido Nítrico Sintase/antagonistas & inibidores , Óxido Nítrico/análise , Pró-Fármacos/farmacologia , Administração Oral , Adulto , Asma/diagnóstico , Asma/enzimologia , Testes Respiratórios , Células Cultivadas , Estudos Cross-Over , Método Duplo-Cego , Inibidores Enzimáticos/administração & dosagem , Feminino , Homoarginina/administração & dosagem , Humanos , Lisina/farmacologia , Masculino , Óxido Nítrico Sintase Tipo II , Pró-Fármacos/administração & dosagem , Mucosa Respiratória/efeitos dos fármacos , Mucosa Respiratória/enzimologia
8.
J Pharmacol Exp Ther ; 304(1): 22-9, 2003 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-12490571

RESUMO

Chronic cellular inflammation and airway wall remodeling with subepithelial fibrosis and airway smooth muscle thickening are features of chronic asthma. We determined the role of nitric oxide in the pathogenesis of allergen-induced airway cell proliferation and inflammation by studying the effects of a relatively selective prodrug inhibitor of nitric-oxide synthase type 2 (NOS2), L-N6-(1-iminoethyl)-lysine-5-tetrazole amide (SC-51). Brown-Norway rats were sensitized to ovalbumin and were exposed to ovalbumin aerosol every 3rd day on six occasions and were treated orally with either vehicle or SC-51 (10 mg. kg(-1); 12 doses). We measured inflammatory cell accumulation in the airways and proliferation of cells by incorporation of bromodeoxyuridine. There was an increase in the total number of airway smooth muscle cells expressing bromodeoxyuridine from 1.3% of airway smooth muscle cells in saline exposed to 5.4% after allergen-exposure (P < 0.001) and airway epithelial cells from 3.3 cells/mm basement membrane to 9.6 after allergen-exposure (P < 0.001). SC-51 had no effect on airway smooth muscle or epithelial cell proliferation. SC-51 attenuated the allergen-induced increase in major basic protein (MBP+) eosinophil (P < 0.05) and CD4+ T-cell (P < 0.05) accumulation. We conclude that nitric oxide derived during allergic inflammation is involved in the expression of eosinophilic inflammation and not in epithelial or airway smooth muscle cell DNA synthesis induced by chronic allergen exposure.


Assuntos
Alérgenos/toxicidade , Homoarginina/análogos & derivados , Inflamação/patologia , Óxido Nítrico/fisiologia , Sistema Respiratório/citologia , Sistema Respiratório/patologia , Actinas/biossíntese , Animais , Bromodesoxiuridina , Líquido da Lavagem Broncoalveolar/citologia , Contagem de Células , Divisão Celular/efeitos dos fármacos , DNA/biossíntese , Inibidores Enzimáticos/farmacologia , Eosinófilos/efeitos dos fármacos , Eosinófilos/metabolismo , Homoarginina/farmacologia , Imunoglobulina E/biossíntese , Imuno-Histoquímica , Inflamação/induzido quimicamente , Isoenzimas/metabolismo , Linfócitos/efeitos dos fármacos , Masculino , Músculo Liso/citologia , Músculo Liso/efeitos dos fármacos , Músculo Liso/patologia , Óxido Nítrico Sintase/antagonistas & inibidores , Óxido Nítrico Sintase Tipo III , Ovalbumina/imunologia , Ratos , Ratos Endogâmicos BN , Sistema Respiratório/efeitos dos fármacos
9.
Exp Eye Res ; 75(5): 521-8, 2002 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-12457864

RESUMO

Following experimental, transient, retinal ischemia in the rat, there is loss of retinal neurons, which occurs over several weeks. Retinal ganglion cells (RGCs) are particularly susceptible and there is early, massive degeneration of these neurons after ischemia. We have determined the early mechanisms by which RGCs are killed following ischemia. Retinal ischemia/reperfusion was produced in rats by transient unilateral elevation of intraocular pressure above systolic blood pressure. Retinas were studied by immunohistochemistry for the presence of inducible nitric oxide synthase (NOS-2) at several time points post-ischemia and specific cell types were identified. Rats were also treated orally with L -N(6) -(1-iminoethyl)lysine 5-tetrazole amide (SC-51), a prodrug of an inhibitor of NOS-2 or with aminoguanidine (AG) for a period of 14 days. Retrograde labelling with Fluoro-Gold quantitated the loss of RGCs. NOS-2 was not present in the normal retina and was not present in the eyes that were contralateral to the ischemic eyes. Within 24hr after ischemia, polymorphonuclear leukocytes containing NOS-2 had entered the ganglion cell layer and surrounded RGCs. Within 5 days after ischemia, NOS-2 was present in many inner retina cells and in invading monocytes in the vitreous. Between 7 and 14 days post-ischemia, there were few hematogenous cells in the retina but NOS-2 was sparsely detectable in microglia and other cells of the inner retina. Two weeks after ischemia, rat eyes lost approximately 50% of the RGCs. Treatment with AG for 14 days following ischemia was partially neuroprotective; approximately 28% of the RGCs were lost. Treatment with SC-51 for 14 days following ischemia almost completely prevented the loss of RGCs. Thus, within 24hr following ischemia, polymorphonuclear leukocytes containing NOS-2 attack and kill neurons in the ganglion cell layer. For 2 weeks after ischemia, NOS-2 appears transiently in the retina in several different cell types at different times. Continuous pharmacological treatment with inhibitors of NOS-2 activity during the 2 weeks post-ischemia period provides significant neuroprotection against the loss of RGCs.


Assuntos
Homoarginina/análogos & derivados , Isquemia/patologia , Óxido Nítrico Sintase/metabolismo , Células Ganglionares da Retina/metabolismo , Vasos Retinianos/metabolismo , Animais , Inibidores Enzimáticos/farmacologia , Guanidinas/farmacologia , Homoarginina/farmacologia , Isquemia/metabolismo , Masculino , Necrose , Neutrófilos/fisiologia , Óxido Nítrico Sintase Tipo II , Ratos , Ratos Endogâmicos F344 , Células Ganglionares da Retina/efeitos dos fármacos , Vasos Retinianos/efeitos dos fármacos
10.
Plant Physiol ; 130(3): 1288-97, 2002 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-12427995

RESUMO

Phytoalexin biosynthesis is part of the defense mechanism of soybean (Glycine max) plants against attack by the fungus Diaporthe phaseolorum f. sp. meridionalis (Dpm), the causal agent of stem canker disease. The treatment of soybean cotyledons with Dpm elicitor or with sodium nitroprusside (SNP), a nitric oxide (NO) donor, resulted in a high accumulation of phytoalexins. This response did not occur when SNP was replaced by ferricyanide, a structural analog of SNP devoid of the NO moiety. Phytoalexin accumulation induced by the fungal elicitor, but not by SNP, was prevented when cotyledons were pretreated with NO synthase (NOS) inhibitors. The Dpm elicitor also induced NOS activity in soybean tissues proximal to the site of inoculation. The induced NOS activity was Ca(2+)- and NADPH-dependent and was sensitive to the NOS inhibitors N(G)-nitro-L-arginine methyl ester, aminoguanidine, and L-N(6)-(iminoethyl) lysine. NOS activity was not observed in SNP-elicited tissues. An antibody to brain NOS labeled a 166-kD protein in elicited and nonelicited cotyledons. Isoflavones (daidzein and genistein), pterocarpans (glyceollins), and flavones (apigenin and luteolin) were identified after exposure to the elicitor or SNP, although the accumulation of glyceollins and apigenin was limited in SNP-elicited compared with fungal-elicited cotyledons. NOS activity preceded the accumulation of these flavonoids in tissues treated with the Dpm elicitor. The accumulation of these metabolites was faster in SNP-elicited than in fungal-elicited cotyledons. We conclude that the response of soybean cotyledons to Dpm elicitor involves NO formation via a constitutive NOS-like enzyme that triggers the biosynthesis of antimicrobial flavonoids.


Assuntos
Fungos/crescimento & desenvolvimento , Glycine max/metabolismo , Homoarginina/análogos & derivados , Óxido Nítrico Sintase/metabolismo , Extratos Vegetais/biossíntese , Sementes/metabolismo , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/farmacologia , Flavonoides/biossíntese , Guanidinas/farmacologia , Homoarginina/farmacologia , NG-Nitroarginina Metil Éster/farmacologia , Doadores de Óxido Nítrico/farmacologia , Óxido Nítrico Sintase/antagonistas & inibidores , Nitroprussiato/farmacologia , Sementes/microbiologia , Sesquiterpenos , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologia , Glycine max/microbiologia , Terpenos , Fitoalexinas
11.
Eur J Pharmacol ; 452(1): 123-33, 2002 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-12323393

RESUMO

The role of nitric oxide (NO) in allergic inflammation and bronchial hyperresponsiveness is unclear. We studied a selective prodrug nitric oxide synthase (NOS)-2 inhibitor, L-N(6)-(1-iminoethyl)lysine 5-tetrazole amide (SC-51). In ovalbumin-sensitized and challenged rats, exhaled NO levels increased by 3 h following challenge (3.73 +/- 0.74 ppb; P < 0.05), peaking at 9 h (11.0 +/- 2.75; P < 0.01) compared to saline controls (1.87 +/- 0.26; P < 0.05 and 2.81 +/- 0.18; P < 0.01). Immunoreactive lung NOS2 expression was increased in ovalbumin-challenged rats compared with ovalbumin-sensitized, saline-challenged rats at 8 h post-challenge. SC-51 (10 mg/kg; p.o.) inhibited allergen-induced increase in exhaled NO levels to 1.3 +/- 0.17 ppb. SC-51 inhibited bronchial hyperresponsiveness in ovalbumin-sensitized and challenged rats (P < 0.05). In sensitized non-exposed rats, SC-51 increased bronchial responsiveness (P < 0.05). SC-51 reduced the allergen-induced increase in bronchoalveolar lavage neutrophils, but caused a nonsignificant reduction in bronchial mucosal eosinophil numbers. NO generated through NOS2 contributes to allergen-induced bronchial hyperresponsiveness but not to bronchial eosinophilia, indicating that these are independently expressed.


Assuntos
Hiper-Reatividade Brônquica/patologia , Homoarginina/análogos & derivados , Hipersensibilidade/patologia , Inflamação/patologia , Óxido Nítrico/fisiologia , Acetilcolina/farmacologia , Resistência das Vias Respiratórias/fisiologia , Animais , Líquido da Lavagem Broncoalveolar/citologia , Contagem de Células , Inibidores Enzimáticos/farmacologia , Eosinófilos/metabolismo , Homoarginina/farmacologia , Imuno-Histoquímica , Isoenzimas/antagonistas & inibidores , Pulmão/patologia , Masculino , Óxido Nítrico Sintase/antagonistas & inibidores , Ovalbumina/imunologia , Ratos , Ratos Endogâmicos BN
12.
J Glaucoma ; 11(3): 221-5, 2002 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-12140399

RESUMO

PURPOSE: To test the hypothesis that nitric oxide, synthesized by inducible nitric oxide synthase, causes degeneration of retinal ganglion cells in an animal model of glaucoma. METHODS: Rats with unilateral, chronic, moderately elevated intraocular pressure were treated orally with L-N(6)-(1-iminoethyl)lysine 5-tetrazole amide, a prodrug of an inhibitor of inducible nitric oxide synthase. The loss of retinal ganglion cells was quantitated as an indicator of glaucomatous damage. RESULTS: At the end of seven months, rat eyes with chronic, moderately elevated intraocular pressure lost approximately 20,000 retinal ganglion cells. Treatment with L-N(6)-(1-iminoethyl)lysine 5-tetrazole amide for seven months completely prevented the loss of retinal ganglion cells in eyes with chronic, moderately elevated intraocular pressure. When treatment with L-N(6)-(1-iminoethyl)lysine 5-tetrazole amide was delayed and started after three months of chronic, moderately elevated intraocular pressure, further loss of retinal ganglion cells was prevented. CONCLUSION: Pharmacological neuroprotection with a selective inhibitor of inducible nitric oxide synthase may be useful for the treatment of glaucoma.


Assuntos
Inibidores Enzimáticos/uso terapêutico , Glaucoma/tratamento farmacológico , Homoarginina/uso terapêutico , Fármacos Neuroprotetores/uso terapêutico , Óxido Nítrico Sintase/antagonistas & inibidores , Pró-Fármacos/uso terapêutico , Administração Oral , Animais , Contagem de Células , Sobrevivência Celular/efeitos dos fármacos , Modelos Animais de Doenças , Homoarginina/análogos & derivados , Pressão Intraocular/efeitos dos fármacos , Masculino , Óxido Nítrico Sintase Tipo II , Hipertensão Ocular/tratamento farmacológico , Ratos , Ratos Endogâmicos BN , Células Ganglionares da Retina/efeitos dos fármacos , Células Ganglionares da Retina/patologia
13.
J Med Chem ; 45(8): 1686-9, 2002 Apr 11.
Artigo em Inglês | MEDLINE | ID: mdl-11931623

RESUMO

The 5-tetrazole amide of L-N(6)-(1-iminoethyl)lysine (L-NIL), L-N(6)-(1-iminoethyl)lysine 5-tetrazole amide (1), has been prepared and evaluated. In contrast to L-NIL, 1 is a stable, nonhygroscopic, crystalline solid. Unlike L-NIL, 1 has minimal inhibitory activity in vitro on human inducible nitric oxide synthase (iNOS). However, it is rapidly converted in vivo to L-NIL and produces dose-dependent inhibition of iNOS in acute and chronic models of inflammation in the rodent with efficacy comparable to L-NIL. In addition, both 1 and L-NIL exhibit significant and comparable in vivo selectivity for the inhibition of iNOS vs endothelial NOS. Doses approximately 80-fold greater than those that inhibited inflammation do not elevate systemic blood pressure. In summary, both the physical properties and the pharmacological profile of 1 make it an ideal molecule for preclinical and clinical studies on the role of selective iNOS inhibitors in mediating inflammatory disease processes.


Assuntos
Inibidores Enzimáticos/síntese química , Homoarginina/síntese química , Lisina/análogos & derivados , Lisina/química , Óxido Nítrico Sintase/antagonistas & inibidores , Pró-Fármacos/síntese química , Doença Aguda , Administração Oral , Animais , Artrite Experimental/patologia , Carragenina , Doença Crônica , Cristalização , Edema/induzido quimicamente , Edema/enzimologia , Edema/patologia , Inibidores Enzimáticos/química , Inibidores Enzimáticos/metabolismo , Homoarginina/análogos & derivados , Homoarginina/química , Homoarginina/metabolismo , Humanos , Isoenzimas/antagonistas & inibidores , Óxido Nítrico Sintase Tipo I , Óxido Nítrico Sintase Tipo II , Óxido Nítrico Sintase Tipo III , Pró-Fármacos/química , Pró-Fármacos/metabolismo , Ratos , Proteínas Recombinantes/antagonistas & inibidores
14.
Cancer Res ; 62(1): 165-70, 2002 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-11782374

RESUMO

The inducible isoforms of nitric oxide synthase (iNOS) and cyclooxygenase (COX-2) are overexpressed in colonic tumors of humans, as well as in colon tumors that develop in rats after the administration of the colon-specific carcinogen, azoxymethane (AOM). iNOS may regulate COX-2 production of proinflammatory prostaglandins, which are known to play a key role in colon tumor development. Experiments were designed to assess the potential chemopreventive properties of highly selective iNOS inhibitors, administered individually and in combination with a selective COX-2 inhibitor, on the development of AOM-induced colonic aberrant crypt foci (ACF). F344 rats were fed experimental diets containing one of the following: 0, 10, 30, or 100 parts/million (ppm) of the selective iNOS inhibitor L-N(6)-(1-iminoethyl)lysine tetrazole-amide (SC-51); 1800 ppm of the less potent, selective iNOS inhibitor aminoguanidine (AG); 500 ppm of the COX-2 inhibitor celecoxib; 320 ppm of the nonsteroidal anti-inflammatory sulindac (positive control); or 30 ppm of SC-51 with 500 ppm of celecoxib, and 100 ppm of SC-51 with 500 ppm of celecoxib. One and 2 weeks later, rats received s.c. injections of AOM at a dose of 15 mg/kg of body weight. At 17 weeks of age, all rats were sacrificed. Colons were evaluated for ACF, and colonic mucosae were assayed for COX and NOS isoform enzyme activities. Samples of venous blood, collected at various time points, were analyzed for these agents. SC-51, administered alone, demonstrated dose-dependent inhibition of the incidence of colonic ACF. The highest doses of SC-51 (100 ppm) and AG (1800 ppm) significantly suppressed the incidence of colonic ACF (P < 0.01 and < 0.001, respectively) and crypt multiplicity in terms of numbers of aberrant crypts/focus (P < 0.0001). Importantly, the combination of either low or high effective doses of SC-51 (30 or 100 ppm) and celecoxib (500 ppm) suppressed AOM-induced colonic ACF formation (P < 0.05 and < 0.001, respectively) and reduced multiplicity of four or more aberrant crypts/focus (P < 0.0001) to a greater extent than did these agents administered individually. As expected, sulindac inhibited colonic ACF formation (P < 0.001) and reduced the multiplicity of four or more aberrant crypts (P < 0.0001) to approximately 45%. The enzymatic activities of COX-2 and iNOS were significantly induced in the AOM-treated animals, and administration of the iNOS inhibitors, SC-51 and AG, significantly inhibited the activities of both iNOS and COX-2 in the colonic mucosa. The combined administration of SC-51 and celecoxib inhibited the COX-2 activity to a greater extent than did either of these agents administered alone. These findings support the hypothesis that selective iNOS inhibitors may have chemopreventive properties and that coadministration with a selective COX-2 inhibitor may have additional chemopreventive potential.


Assuntos
Anticarcinógenos/farmacologia , Neoplasias do Colo/prevenção & controle , Inibidores de Ciclo-Oxigenase/farmacologia , Inibidores Enzimáticos/farmacologia , Homoarginina/farmacologia , Isoenzimas/antagonistas & inibidores , Óxido Nítrico Sintase/antagonistas & inibidores , Sulfonamidas/farmacologia , Animais , Anticarcinógenos/sangue , Azoximetano , Peso Corporal/efeitos dos fármacos , Carcinógenos , Celecoxib , Colo/efeitos dos fármacos , Colo/enzimologia , Neoplasias do Colo/induzido quimicamente , Neoplasias do Colo/enzimologia , Ciclo-Oxigenase 2 , Inibidores de Ciclo-Oxigenase 2 , Inibidores de Ciclo-Oxigenase/sangue , Método Duplo-Cego , Sinergismo Farmacológico , Inibidores Enzimáticos/sangue , Homoarginina/análogos & derivados , Homoarginina/sangue , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/enzimologia , Isoenzimas/biossíntese , Isoenzimas/metabolismo , Masculino , Óxido Nítrico Sintase/metabolismo , Óxido Nítrico Sintase Tipo II , Prostaglandina-Endoperóxido Sintases/biossíntese , Prostaglandina-Endoperóxido Sintases/metabolismo , Pirazóis , Distribuição Aleatória , Ratos , Ratos Endogâmicos F344 , Sulfonamidas/sangue
15.
J Nat Prod ; 63(11): 1566-9, 2000 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-11087612

RESUMO

Lysine is proposed as an alternative biosynthetic precursor of the pyrrole-imidazole alkaloids frequently found in marine sponges. As a putative key intermediate, the natural product Nalpha-(4-bromopyrrolyl-2-carbonyl)-L-homoarginine (1) from the sponge Agelas wiedenmayeri was synthesized in the solid phase starting from Fmoc/Pmc-protected L-homoarginine and in solution starting from readily available L-lysine methyl ester.


Assuntos
Homoarginina/análogos & derivados , Poríferos/química , Alcaloides/metabolismo , Animais , Homoarginina/síntese química , Homoarginina/química , Hidroxilação , Imidazóis/metabolismo , Lisina/química , Lisina/metabolismo , Espectroscopia de Ressonância Magnética , Pirróis/metabolismo , Espectrometria de Massas de Bombardeamento Rápido de Átomos
16.
Eur J Pharmacol ; 216(1): 131-4, 1992 May 27.
Artigo em Inglês | MEDLINE | ID: mdl-1382020

RESUMO

We have assessed the capacity of two novel inhibitors to block cytokine-induced nitric oxide (NO) synthesis by macrophages and vascular smooth muscle cells, as well as NO production by the constitutive enzyme in central nervous system tissue. NG-Cyclopropyl-L-arginine selectively inhibited Ca2+/calmodulin-dependent NO synthesis, with an IC50 of 0.55 microM in brain versus 184 and 258 microM in macrophages and vascular smooth muscle cells, respectively. In contrast, NG-amino-L-homoarginine blocked NO production by all of the cell types examined, with IC50 values ranging from 6.6 to 26 microM. Both inhibitors were active in an in vivo model of endotoxic shock.


Assuntos
Arginina/análogos & derivados , Cerebelo/efeitos dos fármacos , Homoarginina/análogos & derivados , Macrófagos/efeitos dos fármacos , Músculo Liso Vascular/efeitos dos fármacos , Óxido Nítrico/metabolismo , Animais , Arginina/farmacologia , Pressão Sanguínea/efeitos dos fármacos , Cerebelo/metabolismo , Homoarginina/farmacologia , Macrófagos/metabolismo , Masculino , Músculo Liso Vascular/metabolismo , Ratos , Ratos Endogâmicos , Choque Séptico/fisiopatologia
17.
J Med Chem ; 31(1): 65-72, 1988 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-2447279

RESUMO

A new series of unnatural amino acids has been prepared and incorporated into antagonistic analogues of luteinizing hormone-releasing hormone (LH-RH), on the basis of the hypothesis that stabilization of a proposed phospholipid membrane interaction might yield analogues with high potency and a prolonged duration of action. Thus a series of NG,NG'-dialkyl-D-homoarginine analogues [H-D-hArg(R2)-OH; R = Me, Et, Pr, i-Pr, Bu, hexyl, cyclohexyl, (Et, Me2NPr)] was conveniently prepared by semisynthesis from D-Lys using the appropriate dialkylcarbodiimide. A number of the analogues that were prepared by using these new amino acid analogues exhibited very high potency and a prolonged duration of action. One of the most potent members of the series, [N-Ac-D-Nal(2)1,D-pCl-Phe2,D-Trp3,D-hArg(Et2)6,D-Ala10 ]LH-RH (detirelix), had an ED50 of 0.7 microgram in the rat antiovulatory assay when administered at noon on proestrus and only 2.5 micrograms when administered 24 h earlier, at noon on diestrus II. This antagonist is undergoing detailed biological and clinical evaluation.


Assuntos
Arginina/análogos & derivados , Hormônio Liberador de Gonadotropina/análogos & derivados , Hormônio Liberador de Gonadotropina/síntese química , Homoarginina/análogos & derivados , Homoarginina/síntese química , Alquilação , Animais , Hormônio Liberador de Gonadotropina/antagonistas & inibidores , Hormônio Liberador de Gonadotropina/farmacologia , Indicadores e Reagentes , Hormônio Luteinizante/sangue , Masculino , Orquiectomia , Ratos , Ratos Endogâmicos , Relação Estrutura-Atividade
18.
Biochim Biophys Acta ; 576(2): 372-84, 1979 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-427194

RESUMO

Specific modification of the single lysine residue (Lys-12) in glucagon with O-methylisourea has been effected by blocking the reactivity of the amino terminal histidine with copper, providing a method for obtaining [12-homoarginine]glucagon. It was found that as a side reaction, under the conditions of the modification reaction, Cu(II) catalyzed cleavage of the polypeptide chain between Asp-9 and Tyr-10, and between Lys-12 and Tyr-13. This observation may be of value for development of a sequence-specific peptide cleavage procedure. The dilute solution conformations of glucagon and [12-homoarginine]-glucagon were compared by circular dichroism, fluorescence, phosphorescence, energy transfer, and optical detection of magnetic resonance. The results indicate that conversion of Lys-12 to homoarginine does not alter the helix content the side chain conformation in the vicinity of the tyrosine and tryptophan residues, or the relative distances and orientations between these residues. However, the modification reduces the hormone potency towards activation of lipolysis in isolated rat epididymal fat cells by a factor of seven. We attribute the loss of potency to an interference with a specific interaction between the lysine residue and the fat cell hormone receptor, and not to a change in the solution conformation of the hormone.


Assuntos
Glucagon/análogos & derivados , Tecido Adiposo/metabolismo , Sequência de Aminoácidos , Animais , Bioensaio , Dicroísmo Circular , Cobre , Glucagon/farmacologia , Guanidinas , Homoarginina/análogos & derivados , Mobilização Lipídica/efeitos dos fármacos , Masculino , Conformação Proteica , Ratos , Espectrometria de Fluorescência , Suínos
19.
J Biol Chem ; 253(15): 5407-10, 1978 Aug 10.
Artigo em Inglês | MEDLINE | ID: mdl-670205

RESUMO

The alpha-keto acid analogs of arginine and citrulline (and of their next higher homologs, homoarginine and homocitrulline) were prepared enzymatically and shown to exist in equilibrium with cyclic forms which predominate in solution and in the solid state. The cyclic forms of the alpha-keto acid analogs of arginine and homoarginine have the structures pyrrolidine-1-amidino-2-hydroxy-2-carboxylic acid and piperidine-1-amidino-2-hydroxy-2-carboxylic acid, respectively. The cyclic forms of the alpha-keto acid analogs of citrulline and homocitrulline have the structures pyrrolidine-1-carbamyl-2-hydroxy-2-carboxylic acid and piperidine-1-carbamyl-2-carboxylic acid, respectively. The latter compound can undergo further cyclization and dehydration to form additional products, whose structures (2H, 5H, 7H-imidazo[1,5-a]pyridine-1,3-dione and 2H, 5H, 7H, 9H-9-hydroxy-imidazo[1,5-a]pyridine-1,3-dione) were deduced. The alpha-keto acids investigated here, which may be formed reversibly by enzymatic transamination of the corresponding amino acids, may be formed in vivo especially in the presence of metabolic abnormalities associated with inborn enzymatic defects.


Assuntos
Arginina/análogos & derivados , Citrulina/análogos & derivados , Homoarginina/análogos & derivados , Iminoácidos , Cetoácidos , Cinética , Espectroscopia de Ressonância Magnética , Conformação Molecular , Oxirredução , Espectrofotometria , Relação Estrutura-Atividade
20.
Biochem J ; 164(3): 589-94, 1977 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-883953

RESUMO

The biosynthesis of the pyrimidinyl amino acid lathyrine by seedlings of Lathyrus tingitanus L. was shown to be stimulated by uracil. [6(-14)C]Orotate, [2(-14)C]uracil and [3(-14)C]serine were incorporated into lathyrine; the incorporation of [6(-14)C]orotate was substantially decreased in the presence of uracil. Chemical degradation to locate the 14C incorporated from labelled precursors showed that 90% of the radioactivity incorporated into lathyrine from [3(-14)C]serine could be recovered in the alanine side chain. Over 80% of the radioactivity incorporated from [2(-14)C]uracil was shown to be located in C-2 of lathyrine. It is concluded that under the conditions studied, lathyrine arises from a preformed pyrimidine arising via the orotate pathway. Paradoxically, it was also possible to confirm previous reports that radioactivity from L-[guanidino-14C]homoarginine is incorporated into lathyrine and gamma-hydroxyhomoarginine. However, as homoarginine and gamma-hydroxyhomoarginine are also both labelled by [2(-14)C]uracil, it is suggested that they are products of the ring-opening of lathyrine and that reversibility of this process accounts, at least in part, for their observed experimental incorporation into lathyrine.


Assuntos
Diamino Aminoácidos/biossíntese , Plantas/metabolismo , Pirimidinas/biossíntese , Homoarginina/análogos & derivados , Homoarginina/metabolismo , Ácido Orótico/metabolismo , Plantas/efeitos dos fármacos , Uracila/farmacologia
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