RESUMO
We describe practical methods to prepare DOTAGA-DBCO and DFO-DBCO from commercially available starting materials. DOTAGA-DBCO is available in five steps from cyclen with a 33 % overall yield at gram scale. Our synthesis of DFO-DBCO also proceeds in five steps from commercially available starting materials. These bifunctional molecules possess chelating functionality for the binding of medically important radiometals and a strained alkyne suitable for Huisgen cyclization with an azide. These syntheses represent an important step toward improved radioimmunoconjugates for imaging and therapeutic applications.
Assuntos
Ciclamos , Imunoconjugados , Alcinos/química , Azidas/química , Ciclização , Imunoconjugados/farmacologia , Imunoconjugados/uso terapêuticoRESUMO
The orexin system consists of two neuropeptides (orexin-A and orexin-B) that exert their mode of action on two receptors (orexin-1 and orexin-2). While the role of the orexin-2 receptor is established as an important modulator of sleep wake states, the role of the orexin-1 receptor is believed to play a role in addiction, panic, or anxiety. In this manuscript, we describe the optimization of a nonselective substituted azabicyclo[2.2.1]heptane dual orexin receptor antagonist (DORA) into orally bioavailable, brain penetrating, selective orexin-1 receptor (OX1R) antagonists. This resulted in the discovery of our first candidate for clinical development, JNJ-54717793.
RESUMO
We have developed a workflow to extract, separate, and semi-quantify bioactive oxysterols from mouse colon tissues and fecal matters using solid- and liquid-phase extractions, enzymatic and chemical modifications, and stable-isotope dilution LC/MS/MS. The method was applied to a dextran sodium sulphate (DSS)-induced mouse colitis model, which revealed that one particular dihydroxycholesterol (diOHC), 7α,25-diOHC, was significantly elevated in both colon tissue and fecal matters of mice with colitis compared to that in naïve mice. The extent of 7α,25-diOHC elevation was positively correlated with colitis severity.
Assuntos
Colite/induzido quimicamente , Colo/química , Modelos Animais de Doenças , Oxisteróis/isolamento & purificação , Animais , Cromatografia Líquida , Colo/patologia , Sulfato de Dextrana , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Oxisteróis/química , Espectrometria de Massas em TandemRESUMO
Development of a scalable synthesis of an oxazine class of ß-secretase inhibitor is described. Trifluoromethylated acyloin synthesis by the reaction of a mandelic acid with trifluoroacetic anhydride in the presence of pyridine (Dakin-West reaction) was used as an efficient strategy to install the key trifluoromethyl substituent on the oxazine ring. Diastereoselective addition of methyl magnesium bromide to a cyclic sulfamidate imine and trimethylsilyl trifluoromethanesulfonate catalyzed intramolecular amidine formation to yield oxazine-3-amine are some of the significant, novel synthetic methods developed in this synthesis. These critical transformations allowed a concise 11-step route to the target compound with excellent overall yields.
RESUMO
Synthesis of several 7-hydroxy oxysterols and their potential roles as signaling molecules in the innate and adaptive immune responses is discussed. Discovery of a new, fluorinated, synthetic analog of the 7α,25-dihydroxycholesterol-the endogenous ligand of GPR 183 (EBI2), a G-protein coupled receptor highly expressed upon Epstein-Barr virus infection is described. Fluoro oxysterol 12 showed good metabolic stability while maintaining excellent EBI2 agonist activity.
Assuntos
Oxisteróis/química , Animais , Linhagem Celular , Herpesvirus Humano 4/efeitos dos fármacos , Herpesvirus Humano 4/metabolismo , Humanos , Oxisteróis/síntese química , Oxisteróis/farmacologia , Receptores Acoplados a Proteínas G/metabolismoRESUMO
We report the regioselective and direct functionalization of rationally designed imidazole derivatives through electrophilic fluorination with N-fluorobenzenesulfonimide enabled via in situ deprotonation with lithium 2,2,6,6-tetramethylpiperidine. Aided by a controlled protecting group switch, we were able to effectively target both the reactive 5- as well as the difficult to target 4-position of these molecules, leading to a series of fluorinated polysubstituted imidazoles in gram scale.
RESUMO
We report a Cu(I)-catalyzed azide-alkyne-allyl halide three-component reaction for a one-pot synthesis of 1,4-disubstituted 5-allyl-1,2,3-triazoles. The allyl moiety provides not only the electrophile but also a coordinating ligand to Cu, which is essential for the reaction to occur under mild conditions. A concise synthesis of a potential drug candidate 1 is realized based on this key reaction.
RESUMO
The preclinical characterization of novel octahydropyrrolo[3,4-c]pyrroles that are potent and selective orexin-2 antagonists is described. Optimization of physicochemical and DMPK properties led to the discovery of compounds with tissue distribution and duration of action suitable for evaluation in the treatment of primary insomnia. These selective orexin-2 antagonists are proven to promote sleep in rats, and this work ultimately led to the identification of a compound that progressed into human clinical trials for the treatment of primary insomnia. The synthesis, SAR, and optimization of the pharmacokinetic properties of this series of compounds as well as the identification of the clinical candidate, JNJ-42847922 (34), are described herein.
Assuntos
Peptídeos e Proteínas de Sinalização Intracelular/antagonistas & inibidores , Neuropeptídeos/antagonistas & inibidores , Pirróis/química , Pirróis/farmacologia , Animais , Ensaios Clínicos como Assunto , Cães , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Masculino , Neuropeptídeos/metabolismo , Receptores de Orexina/metabolismo , Orexinas , Pirróis/farmacocinética , Pirróis/uso terapêutico , Ratos , Distúrbios do Início e da Manutenção do Sono/tratamento farmacológico , Relação Estrutura-Atividade , Especificidade por SubstratoRESUMO
The preclinical characterization of novel phenyl(piperazin-1-yl)methanones that are histamine H3 receptor antagonists is described. The compounds described are high affinity histamine H3 antagonists. Optimization of the physical properties of these histamine H3 antagonists led to the discovery of several promising lead compounds, and extensive preclinical profiling aided in the identification of compounds with optimal duration of action for wake promoting activity. This led to the discovery of two development candidates for Phase I and Phase II clinical trials.
RESUMO
The RAR-related orphan receptor gamma t (RORγt) is a nuclear receptor required for generating IL-17-producing CD4(+) Th17 T cells, which are essential in host defense and may play key pathogenic roles in autoimmune diseases. Oxysterols elicit profound effects on immune and inflammatory responses as well as on cholesterol and lipid metabolism. Here, we describe the identification of several naturally occurring oxysterols as RORγt agonists. The most potent and selective activator for RORγt is 7ß, 27-dihydroxycholesterol (7ß, 27-OHC). We show that these oxysterols reverse the inhibitory effect of an RORγt antagonist, ursolic acid, in RORγ- or RORγt-dependent cell-based reporter assays. These ligands bind directly to recombinant RORγ ligand binding domain (LBD), promote recruitment of a coactivator peptide, and reduce binding of a corepressor peptide to RORγ LBD. In primary cells, 7ß, 27-OHC and 7α, 27-OHC enhance the differentiation of murine and human IL-17-producing Th17 cells in an RORγt-dependent manner. Importantly, we showed that Th17, but not Th1 cells, preferentially produce these two oxysterols. In vivo, administration of 7ß, 27-OHC in mice enhanced IL-17 production. Mice deficient in CYP27A1, a key enzyme in generating these oxysterols, showed significant reduction of IL-17-producing cells, including CD4(+) and γδ(+) T cells, similar to the deficiency observed in RORγt knockout mice. Our results reveal a previously unknown mechanism for selected oxysterols as immune modulators and a direct role for CYP27A1 in generating these RORγt agonist ligands, which we propose as RORγt endogenous ligands, driving both innate and adaptive IL-17-dependent immune responses.
Assuntos
Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/agonistas , Esteróis/farmacologia , Células Th17/citologia , Animais , Diferenciação Celular , Colestanotriol 26-Mono-Oxigenase/metabolismo , Interleucina-17/biossíntese , Ligantes , Camundongos , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/metabolismo , Esteróis/metabolismoRESUMO
An intramolecular 1,3-dipolar cycloaddition strategy for rapid entry into triazole-fused heterocyclic compounds without recourse to the traditional Cu(1)-catalyzed azide-alkyne cycloadditions is described. Central to the strategy is the in situ generation of substituted diazomethanes in a two-step sequence from the corresponding aldehydes, which then undergo smooth cycloaddition with a cyano group to generate the desired fused 1,2,3-triazoles in good overall yields. The entire sequence can be carried out in a one-pot operation.
Assuntos
Diazometano/química , Compostos Heterocíclicos/síntese química , Nitrilas/química , Triazóis/síntese química , Aldeídos/química , Alcinos/química , Catálise , Cobre/química , Reação de Cicloadição , Compostos Heterocíclicos/química , Estrutura Molecular , Triazóis/químicaRESUMO
Epstein-Barr virus-induced molecule 2 (EBI2) (also known as G-protein-coupled receptor 183) is a G-protein-coupled receptor (GPCR) that is best known for its role in B cell migration and localization. Our recent deorphanization effort led to the discovery of 7α,25-dihydroxycholesterol (7α,25-OHC) as the endogenous ligand for EBI2, which provides a tool for mechanistic studies of EBI2 function. Because EBI2 is the first GPCR known to bind and to be activated by an oxysterol, the goal of this study was to understand the molecular and structural bases for its ligand-dependent activation; this was achieved by identifying structural moieties in EBI2 or in 7α,25-OHC that might affect receptor-ligand interactions. By using a series of chemically related OHC analogs, we demonstrated that all three hydroxyl groups in 7α,25-OHC contributed to ligand-induced activation of the receptor. To determine the location and composition of the ligand binding domain in EBI2, we used a site-directed mutagenesis approach and generated mutant receptors with single amino acid substitutions at selected positions of interest. Biochemical and pharmacological profiling of these mutant receptors allowed for structure-function analyses and revealed critical motifs that likely interact with 7α,25-OHC. By using a hybrid ß(2)-adrenergic receptor-C-X-C chemokine receptor type 4 structure as a template, we created a homology model for EBI2 and optimized the docking of 7α,25-OHC into the putative ligand binding site, so that the hydroxyl groups interact with residues Arg87, Asn114, and Glu183. This model of ligand docking yields important structural insight into the molecular mechanisms mediating EBI2 function and may facilitate future efforts to design novel therapeutic agents that target EBI2.
Assuntos
Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/metabolismo , Sequência de Aminoácidos , Animais , Sítios de Ligação , Células COS , Quimiocinas CXC/genética , Quimiocinas CXC/metabolismo , Chlorocebus aethiops , Cisteína/genética , Cisteína/metabolismo , Humanos , Ligantes , Modelos Moleculares , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida/métodos , Mutação , Receptores Acoplados a Proteínas G/química , Homologia de Sequência de Aminoácidos , Relação Estrutura-AtividadeRESUMO
A POCl(3)-mediated, direct amination reaction of heterocyclic amides/ureas with NH-heterocycles or N-substituted anilines is described. Compared to the existing methods, this operationally simple protocol provides unique reactivity and functional group compatibility because of the metal-free, acidic reaction conditions. The yields are generally excellent.
Assuntos
Amidas/química , Química Farmacêutica/métodos , Compostos Heterocíclicos/síntese química , Medicamentos sob Prescrição/síntese química , Ureia/química , Aminação , Compostos de Anilina/química , Anticolesterolemiantes/análise , Anticolesterolemiantes/química , Compostos Azabicíclicos/análise , Compostos Azabicíclicos/química , Benzamidas , Catálise , Cloridrato de Erlotinib , Zopiclona , Fluorbenzenos/análise , Fluorbenzenos/química , Compostos Heterocíclicos/análise , Humanos , Concentração de Íons de Hidrogênio , Hipnóticos e Sedativos/análise , Hipnóticos e Sedativos/química , Hipoglicemiantes/análise , Hipoglicemiantes/química , Mesilato de Imatinib , Estrutura Molecular , Compostos de Fósforo/química , Piperazinas/análise , Piperazinas/química , Medicamentos sob Prescrição/análise , Inibidores de Proteínas Quinases/análise , Inibidores de Proteínas Quinases/química , Pirimidinas/análise , Pirimidinas/química , Quinazolinas/análise , Quinazolinas/química , Rosiglitazona , Rosuvastatina Cálcica , Sulfonamidas/análise , Sulfonamidas/química , Tiazolidinedionas/análise , Tiazolidinedionas/química , Ureia/análogos & derivadosRESUMO
EBI2 (also called GPR183) is an orphan G-protein-coupled receptor that is highly expressed in spleen and upregulated upon Epstein-Barr-virus infection. Recent studies indicated that this receptor controls follicular B-cell migration and T-cell-dependent antibody production. Oxysterols elicit profound effects on immune and inflammatory responses as well as on cholesterol metabolism. The biological effects of oxysterols have largely been credited to the activation of nuclear hormone receptors. Here we isolate oxysterols from porcine spleen extracts and show that they are endogenous ligands for EBI2. The most potent ligand and activator is 7α,25-dihydroxycholesterol (OHC), with a dissociation constant of 450 pM for EBI2. In vitro, 7α,25-OHC stimulated the migration of EBI2-expressing mouse B and T cells with half-maximum effective concentration values around 500 pM, but had no effect on EBI2-deficient cells. In vivo, EBI2-deficient B cells or normal B cells desensitized by 7α,25-OHC pre-treatment showed reduced homing to follicular areas of the spleen. Blocking the synthesis of 7α,25-OHC in vivo with clotrimazole, a CYP7B1 inhibitor, reduced the content of 7α,25-OHC in the mouse spleen and promoted the migration of adoptively transferred pre-activated B cells to the T/B boundary (the boundary between the T-zone and B-zone in the spleen follicle), mimicking the phenotype of pre-activated B cells from EBI2-deficient mice. Our results show an unexpected causal link between EBI2, an orphan G-protein-coupled receptor controlling B-cell migration, and the known immunological effects of certain oxysterols, thus uncovering a previously unknown role for this class of molecules.
Assuntos
Linfócitos B/efeitos dos fármacos , Hidroxicolesteróis/farmacologia , Receptores Acoplados a Proteínas G/imunologia , Inibidores de 14-alfa Desmetilase/farmacologia , Animais , Linfócitos B/imunologia , Células COS , Linhagem Celular , Movimento Celular/efeitos dos fármacos , Chlorocebus aethiops , Clotrimazol/farmacologia , Humanos , Hidroxicolesteróis/química , Ligantes , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Receptores de Superfície Celular/imunologia , Baço/química , Baço/efeitos dos fármacos , Baço/imunologia , Suínos , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologiaRESUMO
In our pursuit of an efficient, protecting-group-free synthesis of the dual CCK1/CCK2 receptor antagonist 1, we have developed chemoselective conditions for sulfonamide formation reaction in pure water and a PhNMe(2) mediated carboxamide formation, both in the presence of a carboxylic acid. Practical synthesis of an unnatural, chiral ß-aryl-α-amino acid is also described.
Assuntos
Receptor de Colecistocinina A/antagonistas & inibidores , Receptor de Colecistocinina B/antagonistas & inibidores , Estrutura Molecular , EstereoisomerismoRESUMO
We describe a practical and scalable route to compound (Z)-1, a selective CCK1 receptor antagonist. Notable features of this concise route are (1) a regioselective construction of the pyrazole core through the reaction of an aryl hydrazine and an elaborated acetylenic ketone, (2) a Tf2O/pyridine mediated Z-selective dehydration of an α-hydroxyester, and (3) a stereoselective hydrolysis. The sequence is high-yielding and amenable for large-scale synthesis.
Assuntos
Clorobenzenos/síntese química , Dioxóis/síntese química , Dioxóis/farmacologia , Hidrazinas/química , Propionatos/síntese química , Pirazóis/química , Pirazóis/farmacologia , Receptores Acoplados a Proteínas G/antagonistas & inibidores , Clorobenzenos/química , Dioxóis/química , Hidrólise , Cetonas/química , Estrutura Molecular , Propionatos/química , Pirazóis/síntese química , EstereoisomerismoRESUMO
We have recently completed the synthesis of 1-[2-(4-cyclobutyl-[1,4]diazepane-1-carbonyl)-4-(3-fluoro-phenoxy)-pyrrolidin-1-yl]-ethanone, a hydroxyproline-based H(3) receptor antagonist, on 100 g scale. The synthesis proceeds through four steps and route selection was driven by a desire to minimize the cost-of-goods. Naturally occurring trans-4-hydroxy-L-proline was chosen as the precursor to the target's core, which necessitated an inversion at both stereogenic centers. The inversions were accomplished through strategic employment of La Rosa's lactone and a late-stage Mitsunobu reaction. A first generation synthesis that employed N-Boc-homopiperazine was improved in a second generation approach wherein homopiperazine was directly desymmetrized. Finally, the water solubility of a key intermediate necessitated the development of a nonextractive workup for the sodium triacetoxyborohydride reduction.
Assuntos
Azepinas/síntese química , Boroidretos/química , Antagonistas dos Receptores Histamínicos H3/síntese química , Hidroxiprolina/química , Pirrolidinas/síntese química , Sódio/química , Azepinas/química , Cromatografia/métodos , Cromatografia Líquida de Alta Pressão/métodos , Antagonistas dos Receptores Histamínicos H3/química , Espectroscopia de Ressonância Magnética/métodos , Pirrolidinas/química , Solubilidade , Solventes/química , Estereoisomerismo , TemperaturaRESUMO
A series of pyrazole-based thioethers were prepared and found to be potent cathepsin S inhibitors. A crystal structure of 13 suggests that the thioether moiety may bind to the S3 pocket of the enzyme. Additional optimization led to the discovery of aminoethylthioethers with improved enzymatic activity and submicromolar cellular potency.
Assuntos
Catepsinas/antagonistas & inibidores , Catepsinas/metabolismo , Inibidores de Proteases/farmacologia , Pirazóis/farmacologia , Sulfetos/farmacologia , Sítios de Ligação , Catepsinas/química , Linhagem Celular , Cristalografia por Raios X , Humanos , Modelos Moleculares , Inibidores de Proteases/química , Pirazóis/química , Relação Estrutura-Atividade , Sulfetos/químicaRESUMO
A "redox economical" strategy resulted in a concise, modular synthesis of compound 1, a potent Cathepsin S inhibitor. Starting from three building blocks, crude drug substance was prepared in a two-step sequence in high yield. Efficient purification of the crude drug substance was accomplished via the formation of an unusual monoethyl oxalate salt.
Assuntos
Catepsinas/antagonistas & inibidores , Inibidores de Proteases/síntese química , Inibidores de Proteases/farmacologia , Cristalografia por Raios X , Desenho de Fármacos , Espectroscopia de Ressonância Magnética , Modelos Moleculares , Estrutura Molecular , OxirreduçãoRESUMO
CuI/L5 (N,N'-dimethylethylenediamine) proves to be an efficient catalyst system for the amination of arylhalides with guanidines. The same catalyst system is then successfully applied to the one-step synthesis of 1-H-2-amino-benzimidazoles through tandem aminations of 1,2-dihaloarenes in modest yields. This methodology is also applicable for the preparation of 1-H or 1-substutituted 2-aryl- or 2-alkyl-benzimidazoles.