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1.
Pharmacology ; 102(3-4): 169-179, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30099452

RESUMO

In this study, rat and human 11ß-hydroxysteroid dehydrogenase type 1 (11ß-HSD1) have been cloned by lentiviral transduction and expressed by CHO-K1 cells. The results showed that recombinant plasmids contained R11bhsd1 or H11bhsd1 have been constructed, which is consistent with the gene bank respectively. A clone cell was selected with G418 and cultivated to express 11ß-HSD1. 11ß-HSD1 catalytic activity of rat and human were 99.5 and 98.7%, respectively, determined by scanning radiometer. And the cloned CHO-K1 cells expressed the protein of 11ß-HSD1 in a long-term and stable manner, which makes it suitable for screening 11ß-HSD1 inhibitor. The three-dimensional structure of 11ß-HSD1 was used for studying the interaction between inhibitor and enzyme by the binding poses predicted by AutoDock and LeDock software. The docking results revealed that compound 8 forms 2 hydrogen bonds with the residues of Gly-216 and Ile-218 in 11ß-HSD1, that is to say compound 8 maybe a good 11ß-HSD1 inhibitor. Moreover, C57BL/6 mice with R11bHsd1 overexpression had a higher body weight, glucose, total cholesterol, and triglyceride levels compared to the mice treated with an empty viral vector. The results might provide a beneficial foundation for selecting inhibitors of 11ß-HSD1 or for researching drug candidate mechanisms.


Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1/antagonistas & inibidores , Curcumina/análogos & derivados , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/química , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/genética , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/metabolismo , Animais , Células CHO , Clonagem Molecular , Cricetinae , Cricetulus , Curcumina/síntese química , Curcumina/farmacologia , Avaliação Pré-Clínica de Medicamentos/métodos , Inibidores Enzimáticos/síntese química , Lentivirus/genética , Fígado/patologia , Camundongos , Camundongos Endogâmicos C57BL , Simulação de Acoplamento Molecular , Transdução Genética
2.
Eur J Pharmacol ; 835: 169-178, 2018 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-30096297

RESUMO

11ß-hydroxysteroid dehydrogenase type 1 (11ß-HSD1) is a cortisol regenerating enzyme that amplifies tissue glucocorticoid levels, especially in the liver and adipose tissue. Knockout mice or a selective inhibitor of 11ß-HSD1 improves metabolic syndrome parameters in preclinical models and human clinical trials. Here, we evaluated the therapeutic potential of INU-101, a potent and selective oral inhibitor of 11ß-HSD1. The in vitro activity of 11ß-HSD1 was measured using the homogeneous time-resolved fluorescence (HTRF) assay. Differentiated adipocytes were used to evaluate the cellular 11ß-HSD1 activity. To determine the inhibitory effects on 11ß-HSD1 in tissues, we performed ex vivo studies using liver and adipose tissue isolated from C57BL/6 J mice and Cynomolgus monkeys. KKAy mice, ob/ob mice and ZDF rats were administered INU-101 to evaluate whether this compound ameliorated metabolic abnormalities in obese and diabetic animals. INU-101 had highly potent inhibitory activity in mouse, monkey and human 11ß-HSD1, derived from liver microsomes. The oral administration of INU-101 significantly inhibited 11ß-HSD1 activity in the liver and adipose tissue of mice and monkeys. In KKAy mice, ob/ob mice and ZDF rats, the oral administration of INU-101 enhanced insulin sensitivity and lowered the fasting blood glucose level. Furthermore, INU-101 treatment decreased the body weight and ameliorated an improved lipid profile in the diabetic mouse model. These results suggest that the 11ß-HSD1 inhibitor, INU-101 may serve as a novel drug candidate for the treatment of type 2 diabetes and metabolic syndrome.


Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1/antagonistas & inibidores , Adamantano/análogos & derivados , Adamantano/farmacologia , Inibidores Enzimáticos/farmacologia , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/química , Células 3T3-L1 , Adamantano/química , Adamantano/farmacocinética , Animais , Peso Corporal/efeitos dos fármacos , Humanos , Camundongos , Modelos Moleculares , Conformação Proteica , Ratos
3.
Bioorg Chem ; 79: 115-121, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29738970

RESUMO

11ß-Hydroxysteroid dehydrogenase type 1 (11ß-HSD1) is an enzyme that catalyzes the conversion of inactive cortisone into physiologically active cortisol. Inhibiting the activity of this enzyme plays a key role in the treatment of Cushing's syndrome, metabolic syndrome and type 2 diabetes. Therefore, new compounds that are selective inhibitors of this enzyme are constantly being looked for. In this work we present the synthesis of 2-(allylamino)thiazol-4(5H)-one derivatives by the reaction of N-allylthiourea with appropriate α-bromoesters. In the case of using of aliphatic α-bromoesters and α-bromo-ß-phenylesters, the reactions were carried out in a basic medium (sodium ethoxide) and the products were isolated with a yield of up to 68%. Derivatives containing spiro systems in which carbon C-5 of the thiazole ring is the linker atom were obtained in the presence of N,N-diisopropylethylamine. Some of the obtained compounds, at a concentration of 10 µM have activity in the inhibition of 11ß-HSD1 up to 71%. IC50 value for the most active compound: 2-(allylamino)-1-thia-3-azaspiro[4.5]dec-2-en-4-one is 2.5 µM. With a high degree of 11ß-HSD1 inhibition and a relatively large difference in the inhibition of 11ß-HSD1 and 11ß-HSD2 activity, this compound appears to be promising and should be subjected to further testing.


Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Tiazóis/farmacologia , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/química , Sítios de Ligação , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Humanos , Microssomos Hepáticos/metabolismo , Simulação de Acoplamento Molecular , Tiazóis/síntese química , Tiazóis/química
4.
Molecules ; 23(3)2018 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-29495550

RESUMO

We recently found that a cyclohexanecarboxamide derived from 4-azatetracyclo[5.3.2.02,6.08,10]dodec-11-ene displayed low nanomolar inhibition of 11ß-HSD1. In continuation of our efforts to discover potent and selective 11ß-HSD1 inhibitors, herein we explored several replacements for the cyclohexane ring. Some derivatives exhibited potent inhibitory activity against human 11ß-HSD1, although with low selectivity over the isoenzyme 11ß-HSD2, and poor microsomal stability.


Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1/antagonistas & inibidores , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/química , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Técnicas de Química Sintética , Desenho de Drogas , Ativação Enzimática , Inibidores Enzimáticos/síntese química , Humanos , Concentração Inibidora 50 , Espectroscopia de Ressonância Magnética , Relação Estrutura-Atividade
5.
Steroids ; 132: 25-32, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29425740

RESUMO

Many flavonoids and isoflavonoids have anti-diabetic effects in animal models. However, the mechanisms that are involved are generally unclear. Since 11ß-hydroxysteroid dehydrogenases (HSD11Bs) play important roles in diabetes, we hypothesize that flavonoids and isoflavonoids may affect diabetes by targeting two isoforms of HSD11B differently. The inhibitory effects of flavonoids (apigenin and quercetin) and isoflavonoids [genistein and (±) equol] on rat and human HSD11B1 and HSD11B2 were analyzed. The potencies of inhibition on human HSD11B1 reductase was in the order of apigenin > quercetin > genistein > (±) equol, with IC50 values of 2.19, 5.36, 11.00, and over 100 µM, respectively. Genistein also inhibited rat HSD11B1 reductase with IC50 value of 24.58 µM, while other three chemicals showed no effects on the enzyme activity with IC50 values over 100 µM. However, apigenin and (±) equol did not inhibit human HSD11B2 at concentrations as high as 100 µM, while genistein and quercetin inhibited human HSD11B2 by 60% and 50% at 100 µM, respectively. The effective flavonoids and isoflavonoids are noncompetitive inhibitors of HSD11B1 when steroid substrates were used. Docking analysis showed that they bound to the steroid-binding site of the human HSD11B1. These data indicate that apigenin is a selective inhibitor of human HSD11B1 of two HSD11B isoforms, which may be useful in managing symptoms of the metabolic syndrome.


Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1/antagonistas & inibidores , 11-beta-Hidroxiesteroide Desidrogenase Tipo 2/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Flavonoides/farmacologia , Isoflavonas/farmacologia , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/química , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/metabolismo , 11-beta-Hidroxiesteroide Desidrogenase Tipo 2/química , 11-beta-Hidroxiesteroide Desidrogenase Tipo 2/metabolismo , Animais , Apigenina/metabolismo , Apigenina/farmacologia , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/metabolismo , Equol/farmacologia , Flavonoides/metabolismo , Genisteína/farmacologia , Humanos , Isoflavonas/metabolismo , Simulação de Acoplamento Molecular , Conformação Proteica , Quercetina/metabolismo , Quercetina/farmacologia , Ratos
6.
J Comput Aided Mol Des ; 31(7): 603-608, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28623485

RESUMO

Computational scaffold hopping aims to identify core structure replacements in active compounds. To evaluate scaffold hopping potential from a principal point of view, regardless of the computational methods that are applied, a global analysis of conventional scaffolds in analog series from compound activity classes was carried out. The majority of analog series was found to contain multiple scaffolds, thus enabling the detection of intra-series scaffold hops among closely related compounds. More than 1000 activity classes were found to contain increasing proportions of multi-scaffold analog series. Thus, using such activity classes for scaffold hopping analysis is likely to overestimate the scaffold hopping (core structure replacement) potential of computational methods, due to an abundance of artificial scaffold hops that are possible within analog series.


Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1/química , Fator XI/química , Compostos Orgânicos/química , Receptor CB1 de Canabinoide/química , Interpretação Estatística de Dados , Estrutura Molecular , Relação Estrutura-Atividade
7.
J Med Chem ; 60(12): 4932-4948, 2017 06 22.
Artigo em Inglês | MEDLINE | ID: mdl-28537398

RESUMO

BMS-816336 (6n-2), a hydroxy-substituted adamantyl acetamide, has been identified as a novel, potent inhibitor against human 11ß-hydroxysteroid dehydrogenase type 1 (11ß-HSD1) enzyme (IC50 3.0 nM) with >10000-fold selectivity over human 11ß-hydroxysteroid dehydrogenase type 2 (11ß-HSD2). 6n-2 exhibits a robust acute pharmacodynamic effect in cynomolgus monkeys (ED50 0.12 mg/kg) and in DIO mice. It is orally bioavailable (%F ranges from 20 to 72% in preclinical species) and has a predicted pharmacokinetic profile of a high peak to trough ratio and short half-life in humans. This ADME profile met our selection criteria for once daily administration, targeting robust inhibition of 11ß-HSD1 enzyme for the first 12 h period after dosing followed by an "inhibition holiday" so that the potential for hypothalamic-pituitary-adrenal (HPA) axis activation might be mitigated. 6n-2 was found to be well-tolerated in phase 1 clinical studies and represents a potential new treatment for type 2 diabetes, metabolic syndrome, and other human diseases modulated by glucocorticoid control.


Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1/antagonistas & inibidores , Adamantano/análogos & derivados , Azetidinas/farmacologia , Inibidores Enzimáticos/farmacologia , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/química , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/metabolismo , Actinas/antagonistas & inibidores , Adamantano/administração & dosagem , Adamantano/química , Adamantano/farmacologia , Administração Oral , Animais , Azetidinas/administração & dosagem , Azetidinas/química , Disponibilidade Biológica , Cristalografia por Raios X , Cães , Inibidores Enzimáticos/administração & dosagem , Inibidores Enzimáticos/química , Feminino , Meia-Vida , Humanos , Sistema Hipotálamo-Hipofisário/efeitos dos fármacos , Concentração Inibidora 50 , Macaca fascicularis , Masculino , Camundongos Obesos , Ratos , Relação Estrutura-Atividade
8.
Biochim Biophys Acta Gen Subj ; 1861(9): 2342-2353, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28454736

RESUMO

BACKGROUND: Host defence peptides (HDPs) are evolutionarily conserved components of innate immunity. Human HDPs, produced by a variety of immune cells of hematopoietic and epithelial origin, are generally grouped into two families: beta structured defensins and variably-structured cathelicidins. We report the characterization of a very promising cryptic human HDP, here called GVF27, identified in 11-hydroxysteroid dehydrogenase-1 ß-like protein. METHODS: Conformational analysis of GVF27 and its propensity to bind endotoxins were performed by NMR, Circular Dichroism, Fluorescence and Dynamic Light Scattering experiments. Crystal violet and WST-1 assays, ATP leakage measurement and colony counting procedures were used to investigate antimicrobial, anti-biofilm, cytotoxicity and hemolytic activities. Anti-inflammatory properties were evaluated by ELISA. RESULTS: GVF27 possesses significant antibacterial properties on planktonic cells and sessile bacteria forming biofilm, as well as promising dose dependent abilities to inhibit attachment or eradicate existing mature biofilm. It is unstructured in aqueous buffer, whereas it tends to assume a helical conformation in mimic membrane environments as well as it is able to bind lipopolysaccharide (LPS) and lipoteichoic acid (LTA). Notably it is not toxic towards human and murine cell lines and triggers a significant innate immune response by attenuating expression levels of pro-inflammatory interleukins and release of nitric oxide in LPS induced macrophages. CONCLUSION: Human GVF27 may offer significant advantages as leads for the design of human-specific therapeutics. GENERAL SIGNIFICANCE: Human cryptic host defence peptides are naturally no immunogenic and for this they are a real alternative for solving the lack of effective antibiotics to control bacterial infections.


Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1/farmacologia , Anti-Infecciosos/farmacologia , Fragmentos de Peptídeos/farmacologia , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/química , Animais , Anti-Inflamatórios/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Humanos , Lipopolissacarídeos/farmacologia , Espectroscopia de Ressonância Magnética , Camundongos , Fragmentos de Peptídeos/química
10.
Sci Rep ; 6: 26418, 2016 05 19.
Artigo em Inglês | MEDLINE | ID: mdl-27194583

RESUMO

Two new phenone derivatives penicophenones A (1) and B (2), a new cyclic tetrapeptide penicopeptide A (3), and five known compounds were isolated from the culture broth of Penicillium commune, an endophytic fungus derived from Vitis vinifera. Compounds 1-3 were elucidated by extensive spectroscopic analyses including 1D and 2D NMR and HRESIMS. The absolute configurations of 1 and 3 were determined by comparing its ECD with related molecules and modified Marfey's analysis, respectively. Penicophenone A (1) possesses a rare benzannulated 6,6-spiroketal moiety, which is a new member of the unusual structural class with peniphenone A as the representative. Compound 3 exhibited significant inhibition activities against 11ß-hydroxysteroid dehydrogenase type 1 (11ß-HSD1) in vitro and showed strong binding affinity to 11ß-HSD1. Moreover, compound 3 treatments decreased the lipid droplet accumulation associate with the inhibition of 11ß-HSD1 expression in differentiate-induced 3T3-L1 preadipocytes. Furthermore, the molecular docking demonstrated that compound 3 coordinated in the active site of 11ß-HSD1 is essential for the ability of diminishing the enzyme activity.


Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Proteínas Fúngicas/farmacologia , Oligopeptídeos/farmacologia , Penicillium/crescimento & desenvolvimento , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/química , Células 3T3-L1 , Adipócitos/citologia , Adipócitos/efeitos dos fármacos , Adipócitos/metabolismo , Animais , Domínio Catalítico/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Inibidores Enzimáticos/química , Proteínas Fúngicas/química , Humanos , Metabolismo dos Lipídeos/efeitos dos fármacos , Camundongos , Modelos Moleculares , Simulação de Acoplamento Molecular , Estrutura Molecular , Oligopeptídeos/química , Penicillium/metabolismo , Vitis/microbiologia
11.
SAR QSAR Environ Res ; 27(4): 265-92, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27094303

RESUMO

Type 2 diabetes mellitus is described by insulin resistance and high fasting blood glucose. Increased levels of 11ß-hydroxysteroid dehydrogenase type 1 (11ß-HSD1) enzyme result in insulin resistance and metabolic syndrome. Inhibition of 11ß-HSD1 decreases glucose production and increases hepatic insulin sensitivity. Use of selective 11ß-HSD1 inhibitors could prove to be an effective strategy for the treatment of the disease. It was decided to identify the essential structural features required by any compound to possess 11ß-HSD1 inhibitory activity. A dataset of 139 triazoles and tetrazoles having 11ß-HSD1 inhibitory activity was used for the development of a 3D-QSAR model. The best comparative molecular field analysis (CoMFA) model was generated with databased alignment, which was further used for comparative molecular similarity indices analysis (CoMSIA). The optimal CoMSIA model showed [Formula: see text] = 0.809 with five components, [Formula: see text] = 0.931, SEE = 0.323 and F-value = 249.126. The CoMSIA model offered better prediction than the CoMFA model with [Formula: see text] = 0.522 and 0.439, respectively, indicating that the CoMSIA model appeared to be a better one for the prediction of activity for the newly designed 11ß-HSD1 inhibitors. The selectivity aspect of 11ß-HSD1 over 11ß-HSD2 was studied with the help of docking studies.


Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1/antagonistas & inibidores , Simulação de Acoplamento Molecular , Relação Quantitativa Estrutura-Atividade , Tetrazóis/química , Triazóis/química , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/química , Simulação por Computador , Conformação Molecular
12.
Obesity (Silver Spring) ; 23(9): 1856-63, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26239572

RESUMO

OBJECTIVE: Cortisolemia and 11ßHSD1 in liver and adipose tissue are altered in obesity. However, their participation in the development of obesity remains unclear. This study analyzed these parameters in the transition from morbid to type 1 obesity after bariatric surgery. METHODS: A group of 34 patients with morbid obesity and 22 nonobese subjects were recruited. Initial hypothalamus-pituitary-adrenal (HPA) basal activity and 11ßHSD1 mRNA expression in liver, subcutaneous (SAT), and visceral adipose tissue (VAT) were evaluated. A year after bariatric surgery (weight loss of 48 kg), these parameters were reappraised in plasma, SAT, and liver. RESULTS: Body weight loss was accompanied by a downshift in basal HPA activity and 11ßHSD1 expression in SAT. In patients with morbid obesity, 11ßHSD1 expression correlated positively with BMI in VAT and negatively in liver at 6 and 12 months after surgery. In SAT, a correlation was observed with body weight only when patients showed type 1 obesity. Insulin, glucose, and HOMA correlated positively with all the HPA indicators and 11ßHSD1 expression in SAT. CONCLUSIONS: Body weight loss after bariatric surgery is accompanied by a downshift in basal HPA activity. Hepatic and VAT 11ßHSD1 expressions in morbid obesity are predictors of body weight loss.


Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1/metabolismo , Tecido Adiposo/metabolismo , Cirurgia Bariátrica/métodos , Biomarcadores/metabolismo , Hidrocortisona/metabolismo , Gordura Intra-Abdominal/metabolismo , Fígado/metabolismo , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/química , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Obesidade Mórbida/cirurgia , Adulto Jovem
13.
Biochim Biophys Acta ; 1853(7): 1672-82, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25889538

RESUMO

Membrane proteins of the endoplasmic reticulum (ER) are involved in a wide array of essential cellular functions. Identification of the topology of membrane proteins can provide significant insight into their mechanisms of action and biological roles. This is particularly important for membrane enzymes, since their topology determines the subcellular site where a biochemical reaction takes place and the dependence on luminal or cytosolic co-factor pools and substrates. The methods currently available for the determination of topology of proteins are rather laborious and require post-lysis or post-fixation manipulation of cells. In this work, we have developed a simple method for defining intracellular localization and topology of ER membrane proteins in living cells, based on the fusion of the respective protein with redox-sensitive green-fluorescent protein (roGFP). We validated the method and demonstrated that roGFP fusion proteins constitute a reliable tool for the study of ER membrane protein topology, using as control microsomal 11ß-hydroxysteroid dehydrogenase (11ß-HSD) proteins whose topology has been resolved, and comparing with an independent approach. We then implemented this method to determine the membrane topology of six microsomal members of the 17ß-hydroxysteroid dehydrogenase (17ß-HSD) family. The results revealed a luminal orientation of the catalytic site for three enzymes, i.e. 17ß-HSD6, 7 and 12. Knowledge of the intracellular location of the catalytic site of these enzymes will enable future studies on their biological functions and on the role of the luminal co-factor pool.


Assuntos
Retículo Endoplasmático/metabolismo , Proteínas de Fluorescência Verde/metabolismo , Proteínas de Membrana/química , Proteínas de Membrana/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/química , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/metabolismo , 17-Hidroxiesteroide Desidrogenases/química , 17-Hidroxiesteroide Desidrogenases/metabolismo , Permeabilidade da Membrana Celular , Sobrevivência Celular , Simulação por Computador , Células HEK293 , Humanos , Membranas Intracelulares/metabolismo , Modelos Biológicos , Oxirredução , Frações Subcelulares/metabolismo
14.
PLoS One ; 10(2): e0116570, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25659145

RESUMO

MOTIVATION: Using molecular similarity to discover bioactive small molecules with novel chemical scaffolds can be computationally demanding. We describe Ultra-fast Shape Recognition with Atom Types (UFSRAT), an efficient algorithm that considers both the 3D distribution (shape) and electrostatics of atoms to score and retrieve molecules capable of making similar interactions to those of the supplied query. RESULTS: Computational optimization and pre-calculation of molecular descriptors enables a query molecule to be run against a database containing 3.8 million molecules and results returned in under 10 seconds on modest hardware. UFSRAT has been used in pipelines to identify bioactive molecules for two clinically relevant drug targets; FK506-Binding Protein 12 and 11ß-hydroxysteroid dehydrogenase type 1. In the case of FK506-Binding Protein 12, UFSRAT was used as the first step in a structure-based virtual screening pipeline, yielding many actives, of which the most active shows a KD, app of 281 µM and contains a substructure present in the query compound. Success was also achieved running solely the UFSRAT technique to identify new actives for 11ß-hydroxysteroid dehydrogenase type 1, for which the most active displays an IC50 of 67 nM in a cell based assay and contains a substructure radically different to the query. This demonstrates the valuable ability of the UFSRAT algorithm to perform scaffold hops. AVAILABILITY AND IMPLEMENTATION: A web-based implementation of the algorithm is freely available at http://opus.bch.ed.ac.uk/ufsrat/.


Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1 , Algoritmos , Simulação por Computador , Inibidores Enzimáticos , Simulação de Acoplamento Molecular , Proteína 1A de Ligação a Tacrolimo , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/antagonistas & inibidores , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/química , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/metabolismo , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Células HEK293 , Humanos , Proteína 1A de Ligação a Tacrolimo/antagonistas & inibidores , Proteína 1A de Ligação a Tacrolimo/química , Proteína 1A de Ligação a Tacrolimo/metabolismo
15.
Eur J Pharmacol ; 746: 50-5, 2015 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-25445047

RESUMO

To combat the increased morbidity and mortality associated with the developing diabetes epidemic new therapeutic interventions are desirable. Inhibition of intracellular cortisol generation from cortisone by blocking 11ß-hydroxysteroid dehydrogenase 1 (11ß-HSD1) has been shown to ameliorate the risk factors associated with the metabolic syndrome. A challenge in developing 11ß-HSD1 inhibitors has been the species selectivity of small molecules, as many compounds are primate specific. Here we describe our strategy to identify potent selective 11ß-HSD1 inhibitors while ensuring target engagement in key metabolic tissues, liver and fat. This strategy enabled the identification of the clinical candidate, BI 135585.


Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1/antagonistas & inibidores , Diabetes Mellitus Tipo 2/tratamento farmacológico , Inibidores Enzimáticos/farmacologia , Oxazinas/farmacologia , Piridonas/farmacologia , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/química , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/metabolismo , Animais , Domínio Catalítico , Diabetes Mellitus Tipo 2/enzimologia , Avaliação Pré-Clínica de Medicamentos , Inibidores Enzimáticos/química , Inibidores Enzimáticos/uso terapêutico , Humanos , Macaca fascicularis , Masculino , Modelos Moleculares , Oxazinas/química , Oxazinas/uso terapêutico , Piridonas/química , Piridonas/uso terapêutico
16.
J Med Chem ; 57(3): 970-86, 2014 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-24422550

RESUMO

11ß-Hydroxysteroid dehydrogenase type 1 (11ß-HSD1) has been widely considered by the pharmaceutical industry as a target to treat metabolic syndrome in type II diabetics. We hypothesized that central nervous system (CNS) penetration might be required to see efficacy. Starting from a previously reported pyrimidine compound, we removed hydrogen-bond donors to yield 3, which had modest CNS penetration. More significant progress was achieved by changing the core to give 40, which combines good potency and CNS penetration. Compound 40 was dosed to diet-induced obese (DIO) mice and gave excellent target engagement in the liver and high free exposures of drug, both peripherally and in the CNS. However, no body weight reduction or effects on glucose or insulin were observed in this model. Similar data were obtained with a structurally diverse thiazole compound 51. This work casts doubt on the hypothesis that localized tissue modulation of 11ß-HSD1 activity alleviates metabolic syndrome.


Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1/antagonistas & inibidores , Adamantano/análogos & derivados , Adamantano/síntese química , Encéfalo/metabolismo , Diabetes Mellitus Tipo 2/tratamento farmacológico , Hipoglicemiantes/síntese química , Síndrome Metabólica/tratamento farmacológico , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/química , Adamantano/farmacocinética , Adamantano/farmacologia , Animais , Glicemia/metabolismo , Peso Corporal/efeitos dos fármacos , Cristalografia por Raios X , Ciclopropanos/síntese química , Ciclopropanos/farmacocinética , Ciclopropanos/farmacologia , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/fisiopatologia , Gorduras na Dieta/administração & dosagem , Humanos , Hipoglicemiantes/farmacocinética , Hipoglicemiantes/farmacologia , Insulina/sangue , Isoenzimas/antagonistas & inibidores , Isoenzimas/química , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Síndrome Metabólica/metabolismo , Síndrome Metabólica/fisiopatologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Obesos , Modelos Moleculares , Pirazóis/síntese química , Pirazóis/farmacocinética , Pirazóis/farmacologia , Ratos , Estereoisomerismo , Relação Estrutura-Atividade , Tiazóis/síntese química , Tiazóis/farmacocinética , Tiazóis/farmacologia
17.
Mol Cell Endocrinol ; 384(1-2): 71-82, 2014 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-24447464

RESUMO

11 beta-hydroxysteroid dehydrogenase type 1 (11ß-HSD1) converts cortisone to cortisol in a NADPH dependent manner. Overexpression of 11ß-HSD1 in key metabolic tissues is related to the development of type 2 diabetes, obesity, hypertension and metabolic syndrome. Using crystal structures of human 11ß-HSD1 in complex with inhibitors as source of structural information, a combined ligand and structure-based virtual screening approach was implemented to identify novel 11ß-HSD1 inhibitors. A selected group of compounds was identified in silico and further evaluated in cell-based assays for cytotoxicity and 11ß-HSD1 mediated cortisol production inhibitory capacity. The expression of 11ß-HSD1 and 11ß-HSD2 in human LS14 adipocytes was assessed during differentiation. Biological evaluation of 39 compounds in adipocytes and steroids quantification by HPLC-MS/MS identify 4 compounds that exhibit 11ß-HSD1 mediated cortisol production inhibitory activity with potencies in the micromolar range. Two compounds showed to be selective for the 11ß-HSD1 reductase activity and over 11ß-HSD2 isoform, and thus represent novel leads for the development of more active derivatives with higher efficacies targeting intracellular cortisol levels in type 2 diabetes and metabolic syndrome.


Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1/antagonistas & inibidores , 11-beta-Hidroxiesteroide Desidrogenase Tipo 2/antagonistas & inibidores , Adipócitos/efeitos dos fármacos , Inibidores Enzimáticos/química , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/química , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/genética , 11-beta-Hidroxiesteroide Desidrogenase Tipo 2/química , 11-beta-Hidroxiesteroide Desidrogenase Tipo 2/genética , Adipócitos/citologia , Adipócitos/enzimologia , Diferenciação Celular , Linhagem Celular Tumoral , Descoberta de Drogas , Inibidores Enzimáticos/farmacologia , Expressão Gênica , Humanos , Hidrocortisona/antagonistas & inibidores , Hidrocortisona/biossíntese , Cinética , Ligantes , Simulação de Acoplamento Molecular , Relação Estrutura-Atividade , Interface Usuário-Computador
18.
Chem Biol Interact ; 207: 52-7, 2014 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-24246760

RESUMO

Dehydrogenase/reductase (SDR family) member 7 (DHRS7, retSDR4, SDR34C1) is a previously uncharacterized member of the short-chain dehydrogenase/reductase (SDR) superfamily. While human SDR members are known to play an important role in various (patho)biochemical pathways including intermediary metabolism and biotransformation of xenobiotics, only 20% of them are considered to be well characterized. Based on phylogenetic tree and SDR sequence clusters analysis DHRS7 is a close relative to well-known SDR member 11ß-hydroxysteroid dehydrogenase 1 (11ß-HSD1) that participates in metabolism of endogenous and xenobiotic substances with carbonyl group. The aim of present study is to determine the basic biochemical properties of DHRS7 and its possible involvement in metabolism of substrates with carbonyl group. For the first time the computational predictions of this membrane protein and membrane topology were experimentally confirmed. DHRS7 has been demonstrated to be an integral protein facing the lumen of the endoplasmic reticulum with lack of posttranscriptional glycosylation modification. Subsequently, NADP(H) cofactor preference and enzymatic reducing activity of DHRS7 was determined towards endogenous substrates with a steroid structure (cortisone, 4-androstene-3,17-dion) and also toward relevant exogenous substances bearing a carbonyl group harmful to human health (1,2-naphtoquinone, 9,10-phenantrenequinone). In addition to 11ß-HSD1, DHRS7 is another enzyme from SDR superfamily that have been proved, at least in vitro, to contribute to the metabolism of xenobiotics with carbonyl group.


Assuntos
Oxirredutases/metabolismo , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/química , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/metabolismo , Sequência de Aminoácidos , Animais , Benzaldeídos/metabolismo , Western Blotting , Imunofluorescência , Humanos , Membranas Intracelulares/metabolismo , Isoenzimas/química , Isoenzimas/metabolismo , Cinética , Microssomos Hepáticos/enzimologia , Dados de Sequência Molecular , NAD/metabolismo , NADP/metabolismo , Nitrosaminas/química , Nitrosaminas/metabolismo , Oxirredutases/química , Células Sf9 , Espectrofotometria , Especificidade por Substrato , Ultracentrifugação
19.
Org Lett ; 15(22): 5790-3, 2013 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-24175735

RESUMO

An efficient and divergent total synthesis of (+)-colletoic acid, a selective 11-ß hydroxysteroid dehydrogenase 1 (11-ßHSD1) inhibitor, is presented along with its biological activity at the whole-cell level. A scalable, asymmetric synthetic strategy was designed featuring a diversity-oriented synthesis utilizing a diastereoselective intramolecular 5-exo-Heck reaction as the key step to provide the quaternary spirocenter intermediate 9 in multigram scale, thus establishing a platform for further structure-activity relationship studies and providing access to other acorane family members.


Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1/antagonistas & inibidores , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/química , Sesquiterpenos/síntese química , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/metabolismo , Estrutura Molecular , Sesquiterpenos/química , Estereoisomerismo , Relação Estrutura-Atividade
20.
Eur J Pharmacol ; 721(1-3): 70-9, 2013 Dec 05.
Artigo em Inglês | MEDLINE | ID: mdl-24135201

RESUMO

It has been reported that the selective inhibitors of 11ß-hydroxysteroid dehydrogenase type 1 (11ß-HSD1) have considerable potential for treating type 2 diabetes mellitus, metabolic syndrome and inflammation. In the present study, we investigated the anti-diabetic and anti-inflammatory effects of N-(5-carbamoyladamantan-2-yl)-3-((2-fluorophenyl) sulfonyl)thiazolidine-2-carboxamide (KR-67105), a novel 11ß-HSD1 inhibitor, in diabetic mice model and preadipocyte model. KR-67105 concentration dependently inhibited 11ß-HSD1 activity in human and mouse 11ß-HSD1 overexpressing cells and mouse 3T3-L1 adipocytes. Furthermore, KR-67105 concentration-dependently inhibited 11ß-HSD1 activity in the ex vivo assay of C57BL/6 mice. In the study with diet-induced obese (DIO) mice, the administration of KR-67105 (100mg/kg/day, orally for 28 days) improved the glucose tolerance and insulin sensitivity as determined by the oral glucose tolerance test and the insulin tolerance test. Anti-diabetic effect by KR-67105 was associated with the suppression of diabetic related genes expression in liver and fat. Furthermore, KR-67105 suppressed 11ß-HSD1 activity in liver and fat of diabetic mice, but showed no effect on adrenal grand weight/body weight ratio and plasma corticosterone concentration in diabetic mice. In 3T3-L1 preadipocytes, cortisone induced the mRNA of inflammatory cytokines and 11ß-HSD1 and reactive oxygen species formation. This effect was abolished by co-incubation with KR-67105 in a concentration-dependent manner. Moreover, KR-67105 attenuated cortisone induced iNOS expression and phosphorylation of NF-κB p65, p38 MAPK, and ERK1/2 in preadipocytes. Taken together, it is concluded that a selective 11ß-HSD1 inhibitor, KR-67105, may provide a new therapeutic window in the prevention and treatment of type 2 diabetes with chronic inflammation without toxicity.


Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1/antagonistas & inibidores , Anti-Inflamatórios/farmacologia , Dieta/efeitos adversos , Hipoglicemiantes/farmacologia , Obesidade/tratamento farmacológico , Tiazolidinas/farmacologia , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/química , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/metabolismo , Células 3T3-L1 , Adipócitos/efeitos dos fármacos , Adipócitos/metabolismo , Adipócitos/patologia , Animais , Anti-Inflamatórios/metabolismo , Anti-Inflamatórios/uso terapêutico , Cortisona/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Teste de Tolerância a Glucose , Humanos , Hipoglicemiantes/metabolismo , Hipoglicemiantes/uso terapêutico , Hipotálamo/efeitos dos fármacos , Hipotálamo/fisiopatologia , Resistência à Insulina , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Camundongos , Camundongos Obesos , Modelos Moleculares , Obesidade/metabolismo , Obesidade/patologia , Obesidade/fisiopatologia , Sistema Hipófise-Suprarrenal/efeitos dos fármacos , Sistema Hipófise-Suprarrenal/fisiopatologia , Conformação Proteica , Tiazolidinas/metabolismo , Tiazolidinas/uso terapêutico
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