RESUMO
To further explore the optimum placement of the acid moiety in conformationally constrained analogs of GW 4064 1a, a series of stilbene replacements were prepared. The benzothiophene 1f and the indole 1g display the optimal orientation of the carboxylate for enhanced FXR agonist potency.
Assuntos
Isoxazóis/química , Isoxazóis/farmacologia , Receptores Citoplasmáticos e Nucleares/agonistas , Estilbenos/química , Estilbenos/farmacologia , Sequência de Aminoácidos , Animais , Linhagem Celular , Humanos , Conformação Molecular , Dados de Sequência Molecular , Receptores Citoplasmáticos e Nucleares/metabolismoRESUMO
To improve on the drug properties of GSK8062 1b, a series of heteroaryl bicyclic naphthalene replacements were prepared. The quinoline 1c was an equipotent FXR agonist with improved drug developability parameters relative to 1b. In addition, analog 1c lowered body weight gain and serum glucose in a DIO mouse model of diabetes.
Assuntos
Isoxazóis/química , Naftalenos/química , Quinolinas/química , Receptores Citoplasmáticos e Nucleares/agonistas , Animais , Sítios de Ligação , Glicemia/metabolismo , Cristalografia por Raios X , Diabetes Mellitus Experimental/metabolismo , Cães , Transferência Ressonante de Energia de Fluorescência , Humanos , Isoxazóis/síntese química , Isoxazóis/farmacocinética , Ligantes , Camundongos , Conformação Molecular , Estrutura Terciária de Proteína , Quinolinas/síntese química , Quinolinas/farmacocinética , Ratos , Receptores Citoplasmáticos e Nucleares/metabolismo , Aumento de Peso/efeitos dos fármacosRESUMO
Inhibition of acetyl-CoA carboxylase (ACC) may prevent lipid-induced insulin resistance and type 2 diabetes, making the enzyme an attractive pharmaceutical target. Although the enzyme is highly conserved amongst animals, only the yeast enzyme structure is available for rational drug design. The use of biophysical assays has permitted the identification of a specific C-terminal truncation of the 826-residue human ACC2 carboxyl transferase (CT) domain that is both functionally competent to bind inhibitors and crystallizes in their presence. This C-terminal truncation led to the determination of the human ACC2 CT domain-CP-640186 complex crystal structure, which revealed distinctions from the yeast-enzyme complex. The human ACC2 CT-domain C-terminus is comprised of three intertwined alpha-helices that extend outwards from the enzyme on the opposite side to the ligand-binding site. Differences in the observed inhibitor conformation between the yeast and human structures are caused by differing residues in the binding pocket.
Assuntos
Acetil-CoA Carboxilase/química , Acetil-CoA Carboxilase/antagonistas & inibidores , Acetil-CoA Carboxilase/isolamento & purificação , Acetil-CoA Carboxilase/metabolismo , Acetiltransferases/antagonistas & inibidores , Sequência de Aminoácidos , Sítios de Ligação , Dicroísmo Circular , Cristalografia por Raios X , Ácidos Graxos/metabolismo , Polarização de Fluorescência , Humanos , Ligantes , Modelos Moleculares , Dados de Sequência Molecular , Conformação Proteica , Desnaturação Proteica , Estrutura Terciária de Proteína , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/isolamento & purificação , Proteínas Recombinantes de Fusão/metabolismo , Proteínas de Saccharomyces cerevisiae/química , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Especificidade da Espécie , Relação Estrutura-AtividadeRESUMO
Starting from the known FXR agonist GW 4064 1a, a series of alternately 3,5-substituted isoxazoles was prepared. Several of these analogs were potent full FXR agonists. A subset of this series, with a tether between the isoxazole ring and the 3-position aryl substituent, were equipotent FXR agonists to GW 4064 1a, with the 2,6-dimethyl phenol analog 1t having greater FRET FXR potency than GW 4064 1a.
Assuntos
Isoxazóis/síntese química , Receptores Citoplasmáticos e Nucleares/agonistas , Animais , Cristalografia por Raios X , Transferência Ressonante de Energia de Fluorescência , Isoxazóis/química , Isoxazóis/farmacologia , Ratos , Receptores Citoplasmáticos e Nucleares/metabolismo , Relação Estrutura-AtividadeRESUMO
Two series of conformationally constrained analogs of the FXR agonist GW 4064 1 were prepared. Replacement of the metabolically labile stilbene with either benzothiophene or naphthalene rings led to the identification of potent full agonists 2a and 2g.
Assuntos
Isoxazóis/química , Naftalenos/química , Receptores Citoplasmáticos e Nucleares/agonistas , Tiofenos/química , Animais , Sítios de Ligação , Simulação por Computador , Cristalografia por Raios X , Humanos , Isoxazóis/farmacologia , Naftalenos/farmacocinética , Ratos , Receptores Citoplasmáticos e Nucleares/metabolismo , Relação Estrutura-Atividade , Tiofenos/farmacocinéticaRESUMO
Explorations of the S(1') subsite of ACE2 via modifications of the P(1') methylene biphenyl moiety of thiol-based metalloprotease inhibitors led to improvements in ACE2 selectivity versus ACE and NEP, while maintaining potent ACE2 inhibition.
Assuntos
Inibidores da Enzima Conversora de Angiotensina/química , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Peptidil Dipeptidase A/metabolismo , Compostos de Sulfidrila/química , Compostos de Sulfidrila/farmacologia , Enzima de Conversão de Angiotensina 2 , Sítios de Ligação , Humanos , Modelos Moleculares , Estrutura Molecular , Proteínas Recombinantes , Relação Estrutura-AtividadeRESUMO
Screening of a metalloprotease library led to the identification of a thiol-based dual ACE/NEP inhibitor as a potent ACE2 inhibitor. Modifications of the P(1) benzyl moiety led to improvements in ACE2 potency as well as to increased selectivity versus ACE and NEP.
Assuntos
Inibidores da Enzima Conversora de Angiotensina/química , Peptidil Dipeptidase A/efeitos dos fármacos , Compostos de Sulfidrila/química , Enzima de Conversão de Angiotensina 2 , Inibidores da Enzima Conversora de Angiotensina/metabolismo , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Animais , Feminino , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Peptidil Dipeptidase A/genética , Relação Estrutura-Atividade , Especificidade por Substrato , Compostos de Sulfidrila/metabolismo , Compostos de Sulfidrila/farmacologiaRESUMO
A series of estrogen receptor ligands based on a 3-alkyl naphthalene scaffold was synthesized using an intramolecular enolate-alkyne cycloaromatization as the key step. Several of these compounds bearing a C6-OH group were shown to be high affinity ligands. All compounds had similar ERalpha and ERbeta binding affinity ranging from micromolar to low nanomolar.
Assuntos
Naftalenos/síntese química , Naftalenos/farmacologia , Receptores de Estrogênio/agonistas , Técnicas de Química Combinatória , Moduladores de Receptor Estrogênico/metabolismo , Receptor alfa de Estrogênio/metabolismo , Receptor beta de Estrogênio/metabolismo , Feminino , Humanos , Ligantes , Estrutura Molecular , Naftalenos/química , Moduladores Seletivos de Receptor Estrogênico/metabolismo , Relação Estrutura-AtividadeRESUMO
We report the synthesis and in vitro activity of a series of novel pyrrolidinyl pyridones and pyrazinones as potent inhibitors of prolyl oligopeptidase (POP). Within this series, compound 39 was co-crystallized within the catalytic site of a human chimeric POP protein which provided a more detailed understanding of how these inhibitors interacted with the key residues within the catalytic pocket.
Assuntos
Piridonas/síntese química , Piridonas/farmacologia , Pirrolidinas/síntese química , Pirrolidinas/farmacologia , Serina Endopeptidases/efeitos dos fármacos , Inibidores de Serina Proteinase/síntese química , Inibidores de Serina Proteinase/farmacologia , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Técnicas de Química Combinatória , Cristalografia por Raios X , Desenho de Fármacos , Humanos , Conformação Molecular , Estrutura Molecular , Prolil Oligopeptidases , Piridonas/sangue , Piridonas/química , Pirrolidinas/sangue , Pirrolidinas/química , Inibidores de Serina Proteinase/sangue , Inibidores de Serina Proteinase/químicaRESUMO
Starting from the known FXR agonist GW 4064 1a, a series of stilbene replacements were prepared. The 6-substituted 1-naphthoic acid 1b was an equipotent FXR agonist with improved developability parameters relative to 1a. Analog 1b also reduced the severity of cholestasis in the ANIT acute cholestatic rat model.
Assuntos
Ácidos Carboxílicos/síntese química , Ácidos Carboxílicos/farmacologia , Colestase/tratamento farmacológico , Proteínas de Ligação a DNA/agonistas , Isoxazóis/síntese química , Isoxazóis/farmacologia , Naftalenos/síntese química , Naftalenos/farmacologia , Receptores Citoplasmáticos e Nucleares/agonistas , Fatores de Transcrição/agonistas , Administração Oral , Animais , Ácidos Carboxílicos/química , Colestase/metabolismo , Colestase/patologia , Cristalografia por Raios X , Proteínas de Ligação a DNA/química , Modelos Animais de Doenças , Cães , Mucosa Gástrica/metabolismo , Haplorrinos , Isoxazóis/química , Camundongos , Conformação Molecular , Estrutura Molecular , Naftalenos/química , Ratos , Receptores Citoplasmáticos e Nucleares/química , Estômago/efeitos dos fármacos , Relação Estrutura-Atividade , Fatores de Transcrição/químicaRESUMO
The estrogen-related receptor alpha (ERRalpha) is a potential target for activation in the treatment of metabolic disease. To date, no small-molecule agonists of ERRalpha have been identified despite several high-throughput screening campaigns. We describe the synthesis and profiling of a small array of compounds designed on the basis of a previously reported agonist-bound crystal structure of the closely related receptor ERRgamma. The results suggest that ERRalpha may be intractable as a direct target for pharmacologic activation.
Assuntos
Hidrazonas/química , Hidrazonas/farmacologia , Receptores de Estrogênio/agonistas , Receptores de Estrogênio/química , Sítios de Ligação , Cristalografia por Raios X , Agonismo Inverso de Drogas , Transferência Ressonante de Energia de Fluorescência , Células HeLa , Humanos , Hidrazonas/síntese química , Receptores de Estrogênio/antagonistas & inibidores , Receptor ERRalfa Relacionado ao EstrogênioRESUMO
The 2.7 A X-ray crystal structure of the HNF4gamma ligand binding domain (LBD) revealed the presence of a fatty acid within the pocket, with the AF2 helix in a conformation characteristic of a transcriptionally active nuclear receptor. GC/MS and NMR analysis of chloroform/methanol extracts from purified HNF4alpha and HNF4gamma LBDs identified mixtures of saturated and cis-monounsaturated C14-18 fatty acids. The purified HNF4 LBDs interacted with nuclear receptor coactivators, and both HNF4 subtypes show high constitutive activity in transient transfection assays, which was reduced by mutations designed to interfere with fatty acid binding. The endogenous fatty acids did not readily exchange with radiolabeled palmitic acid, and all attempts to displace them without denaturing the protein failed. Our results suggest that the HNF4s may be transcription factors that are constitutively bound to fatty acids.
Assuntos
Proteínas de Ligação a DNA , Ácidos Graxos/metabolismo , Fosfoproteínas/química , Fosfoproteínas/metabolismo , Fatores de Transcrição/química , Fatores de Transcrição/metabolismo , Sequência de Aminoácidos , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos , Linhagem Celular , Cristalografia por Raios X , Cromatografia Gasosa-Espectrometria de Massas , Fator 4 Nuclear de Hepatócito , Humanos , Espectroscopia de Ressonância Magnética , Modelos Moleculares , Dados de Sequência Molecular , Ligação Proteica , Conformação Proteica , Isoformas de Proteínas/química , Homologia de Sequência de Aminoácidos , Relação Estrutura-Atividade , Transcrição GênicaRESUMO
The first small molecule agonists of the estrogen-related receptors have been identified. GSK4716 (3) and GSK9089 (4) show binding to ERRgamma with remarkable selectivity over the classical estrogen receptors. Notably, in cell-based reporter assays, 3 mimics the protein ligand PGC-1alpha in activation of human ERRbeta and ERRgamma.
Assuntos
Receptor beta de Estrogênio/agonistas , Hidrazinas/síntese química , Fenóis/síntese química , Receptores de Estrogênio/agonistas , Ligação Competitiva , Receptor beta de Estrogênio/genética , Transferência Ressonante de Energia de Fluorescência , Genes Reporter , Células HeLa , Proteínas de Choque Térmico/biossíntese , Humanos , Hidrazinas/química , Hidrazinas/farmacologia , Ligantes , Mimetismo Molecular , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo , Fenóis/química , Fenóis/farmacologia , Receptores de Estrogênio/genética , Relação Estrutura-Atividade , Fatores de Transcrição/biossínteseRESUMO
Cathepsin K, a lysosomal cysteine protease of the papain superfamily, is abundantly and selectively expressed in osteoclasts, suggesting that this enzyme is crucial for bone resorption. Prevention of osteoclast-mediated bone resorption via inhibition of cathepsin K could be an effective approach to prevent osteoporosis. Potent and selective reversible ketoamide-based inhibitors have been identified in the present study. Using a known crystal structure of a ketoamide-based inhibitor, information from residues that form the P2/P3 pocket was used in the design of inhibitors that could allow for gains in selectivity and potency. Further, incorporation of P' selective heterocycles, along with the P2/P3 modifications, is also described. These modifications have resulted in potent and selective cathepsin K inhibitors that allow for improvements in their physiochemical properties and represent a viable lead series for the discovery of new therapies for the prevention and treatment of osteoporosis
Assuntos
Amidas/síntese química , Catepsinas/antagonistas & inibidores , Cetonas/síntese química , Amidas/química , Carbamatos/síntese química , Carbamatos/química , Catepsina K , Catepsinas/química , Ciclobutanos/síntese química , Ciclobutanos/química , Ciclopentanos/síntese química , Ciclopentanos/química , Cetonas/química , Modelos Moleculares , Pirazóis/síntese química , Pirazóis/química , Relação Estrutura-AtividadeRESUMO
A series of tetrahydrobenzofuranyl and tetrahydrobenzothienyl propenoic acids that showed potent agonist activity against RXRalpha were synthesized via a structure-based design approach. Among the compounds studied, 46a,b showed not only very good potency against RXRalpha (K(i) = 6 nM) but was also found to be greater than 167-fold selective vs RARalpha (K(i) > 1000 nM). This compound profiled out as a full agonist in a cell-based transient transfection assay (EC(50) = 3 nM). The two antipodes were separated via chiral chromatography, and 46b was found to be 40-fold more potent than 46a. Interestingly, cocrystallization of 46a,b with the RXRalpha protein generated a liganded structure whereby the (S)-antipode was found in the binding pocket. Given orally in db/db mice or ZDF rats, 46a,b showed a significant glucose-lowering effect and an increase in liver mass. Triglycerides decreased significantly in db/db mice but increased in the ZDF rats. A dose-dependent decrease of nonesterified free fatty acids was seen in ZDF rats but not in db/db mice. These differences indicate a species specific effect of RXR agonists on lipid metabolism.
Assuntos
Acrilatos/síntese química , Benzofuranos/síntese química , Hipoglicemiantes/síntese química , Receptores do Ácido Retinoico/agonistas , Fatores de Transcrição/agonistas , Acrilatos/química , Acrilatos/farmacologia , Animais , Benzofuranos/química , Benzofuranos/farmacologia , Sítios de Ligação , Linhagem Celular , Cristalografia por Raios X , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/tratamento farmacológico , Haplorrinos , Humanos , Hipoglicemiantes/química , Hipoglicemiantes/farmacologia , Ligantes , Lipídeos/biossíntese , Masculino , Camundongos , Modelos Moleculares , Ensaio Radioligante , Ratos , Ratos Zucker , Receptores do Ácido Retinoico/química , Receptores do Ácido Retinoico/genética , Receptores X de Retinoides , Estereoisomerismo , Relação Estrutura-Atividade , Fatores de Transcrição/química , Fatores de Transcrição/genética , TransfecçãoRESUMO
Osteoclast-mediated bone matrix resorption has been attributed to cathepsin K, a cysteine protease of the papain family that is abundantly and selectively expressed in osteoclast. Inhibition of cathepsin K could potentially be an effective method to prevent osteoporosis. Structure-activity studies on a series of reversible ketoamides based inhibitors of cathepsin K have led to identification of potent and selective compounds. Crystallographic studies have given insights into the mode of binding of these inhibitors. A series of ketoamides with varying P1 moieties were first synthesized to find an optimum group that would fit into the S1 subsite of the cysteine protease, cathepsin K. With a desired P1 group in place a variety of heterocyclic analogues in the P' region were synthesized to study their steric and electronic effects. In the process of exploring these P' heterocyclic variations, excellent selectivity was gained over other highly homologous cysteine proteases, including cathepsins L, S, and V. The favorable pharmacokinetic properties of some of these cathepsin K inhibitors in rats make them suitable for evaluation in rodent osteoporosis models. A representative cathepsin K inhibitor was shown to attenuate PTH-stimulated hypercalcemia in the TPTX rat model. These inhibitors provide a viable lead series in the discovery of new therapies for the prevention and treatment of osteoporosis
Assuntos
Amidas/síntese química , Catepsinas/antagonistas & inibidores , Inibidores de Cisteína Proteinase/síntese química , Cetonas/síntese química , Administração Oral , Amidas/farmacocinética , Amidas/farmacologia , Animais , Disponibilidade Biológica , Cálcio/sangue , Catepsina K , Catepsinas/química , Cristalografia por Raios X , Inibidores de Cisteína Proteinase/farmacocinética , Inibidores de Cisteína Proteinase/farmacologia , Humanos , Cetonas/farmacocinética , Cetonas/farmacologia , Masculino , Modelos Moleculares , Estrutura Molecular , Osteoporose/metabolismo , Ratos , Ratos Wistar , Proteínas Recombinantes/antagonistas & inibidores , Proteínas Recombinantes/química , Relação Estrutura-AtividadeRESUMO
Starting from a potent ketone-based inhibitor with poor drug properties, incorporation of P(2)-P(3) elements from a ketoamide-based inhibitor led to the identification of a hybrid series of ketone-based cathepsin K inhibitors with better oral bioavailability than the starting ketone.
Assuntos
Catepsinas/antagonistas & inibidores , Inibidores de Cisteína Proteinase/química , Inibidores de Cisteína Proteinase/farmacocinética , Cetonas/química , Cetonas/farmacocinética , Administração Oral , Disponibilidade Biológica , Catepsina K , Catepsinas/química , Cristalografia por Raios X , Inibidores de Cisteína Proteinase/administração & dosagem , Humanos , Cetonas/administração & dosagem , Conformação Proteica , Relação Estrutura-AtividadeRESUMO
X-ray crystal structures of the ligand binding domain (LBD) of the estrogen-related receptor-gamma (ERRgamma) were determined that describe this receptor in three distinct states: unliganded, inverse agonist bound, and agonist bound. Two structures were solved for the unliganded state, the ERRgamma LBD alone, and in complex with a coregulator peptide representing a portion of receptor interacting protein 140 (RIP140). No significant differences were seen between these structures that both exhibited the conformation of ERRgamma seen in studies with other coactivators. Two structures were obtained describing the inverse agonist-bound state, the ERRgamma LBD with 4-hydroxytamoxifen (4-OHT), and the ERRgamma LBD with 4-OHT and a peptide representing a portion of the silencing mediator of retinoid and thyroid hormone action protein (SMRT). The 4-OHT structure was similar to other reported inverse agonist bound structures, showing reorientation of phenylalanine 435 and a displacement of the AF-2 helix relative to the unliganded structures with little other rearrangement occurring. No significant changes to the LBD appear to be induced by peptide binding with the addition of the SMRT peptide to the ERRgamma plus 4-OHT complex. The observed agonist-bound state contains the ERRgamma LBD, a ligand (GSK4716), and the RIP140 peptide and reveals an unexpected rearrangement of the phenol-binding residues. Thermal stability studies show that agonist binding leads to global stabilization of the ligand binding domain. In contrast to the conventional mechanism of nuclear receptor ligand activation, activation of ERRgamma by GSK4716 does not appear to involve a major rearrangement or significant stabilization of the C-terminal helix.
Assuntos
Receptores Citoplasmáticos e Nucleares/química , Receptores de Estrogênio/química , Sítios de Ligação , Dicroísmo Circular , Cristalografia por Raios X , Humanos , Técnicas In Vitro , Ligantes , Modelos Moleculares , Complexos Multiproteicos , Estrutura Quaternária de Proteína , Estrutura Terciária de Proteína , Receptores Citoplasmáticos e Nucleares/agonistas , Receptores Citoplasmáticos e Nucleares/genética , Receptores Citoplasmáticos e Nucleares/metabolismo , Receptores de Estrogênio/agonistas , Receptores de Estrogênio/genética , Receptores de Estrogênio/metabolismo , Proteínas Recombinantes/agonistas , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Eletricidade EstáticaRESUMO
The design and synthesis of 4-hydroxytamoxifen (4-OHT) derivatives are described. The binding affinities of these compounds toward the orphan estrogen-related receptor gamma and the classical estrogen receptor alpha demonstrate that analogs bearing hydroxyalkyl groups display improved binding selectivity profiles compared with that of 4-OHT. An X-ray crystal structure of one of the designed compounds bound to ERRgamma LBD confirms the molecular basis of the selectivity.
Assuntos
Receptores Citoplasmáticos e Nucleares/efeitos dos fármacos , Receptores de Estrogênio/efeitos dos fármacos , Tamoxifeno , Sítios de Ligação/efeitos dos fármacos , Cristalografia por Raios X , Desenho de Fármacos , Receptor alfa de Estrogênio/efeitos dos fármacos , Humanos , Ligantes , Modelos Moleculares , Estrutura Molecular , Relação Estrutura-Atividade , Tamoxifeno/análogos & derivados , Tamoxifeno/síntese química , Tamoxifeno/farmacologiaRESUMO
Starting from a potent pantolactone ketoamide cathepsin K inhibitor discovered from structural screening, conversion of the lactone scaffold to a pyrrolidine scaffold allowed exploration of the S(3) subsite of cathepsin K. Manipulation of P3 and P1' groups afforded potent inhibitors with drug-like properties.