RESUMEN
FTY720 is an immunosuppressive agent that modulates lymphocyte trafficking. It is phosphorylated in vivo to FTY720-phosphate (FTY-P) and binds to a family of G protein-coupled receptors recognizing sphingosine 1-phosphate (S1P) as the natural ligand. It has previously been reported that FTY-P blocks egress of lymphocytes from the thymus and lymph nodes, resulting in peripheral blood lymphopenia. We now report that FTY-P also causes displacement of marginal zone (MZ) B cells to the splenic follicles, an effect that is similar to that observed after in vivo administration of lipopolysaccharide. This effect is specific to B cells in the MZ, as treatment with FTY-P does not cause redistribution of the resident macrophage population. A small but statistically significant decrease in the expression of beta1 integrin on MZ B cells was observed with FTY-P treatment. The redistribution of MZ B cells from the MZ sinuses does not abolish the ability of these cells to respond to the T-independent antigen, trinitrophenol-Ficoll. It has been proposed that the displacement of MZ B cells to the follicles is an indication of cell activation. Consistent with this, FTY-P caused an increase in percentage of MZ B cells expressing activation markers CD9, CD1d, and CD24. These results suggest that S1P receptors on MZ B cells are responsible for their mobilization to follicles.
Asunto(s)
Linfocitos B/fisiología , Movimiento Celular/efectos de los fármacos , Centro Germinal/metabolismo , Inmunosupresores/administración & dosificación , Glicoles de Propileno/administración & dosificación , Receptores de Lisoesfingolípidos/antagonistas & inhibidores , Animales , Antígenos CD/biosíntesis , Linfocitos B/citología , Movimiento Celular/fisiología , Femenino , Clorhidrato de Fingolimod , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/fisiología , Centro Germinal/citología , Inyecciones Intraperitoneales , Lipopolisacáridos/administración & dosificación , Activación de Linfocitos/efectos de los fármacos , Activación de Linfocitos/fisiología , Tejido Linfoide/citología , Tejido Linfoide/metabolismo , Macrófagos/citología , Macrófagos/metabolismo , Ratones , Receptores de Lisoesfingolípidos/metabolismo , Esfingosina/análogos & derivadosRESUMEN
The discovery of novel 4-hydroxy-2-(heterocyclic)pyrimidine-5-carboxamide inhibitors of hypoxia-inducible factor (HIF) prolyl hydroxylases (PHD) is described. These are potent, selective, orally bioavailable across several species, and active in stimulating erythropoiesis. Mouse and rat studies showed hematological changes with elevations of plasma EPO and circulating reticulocytes following single oral dose administration, while 4-week q.d. po administration in rat elevated hemoglobin levels. A major focus of the optimization process was to decrease the long half-life observed in higher species with early compounds. These efforts led to the identification of 28 (MK-8617), which has advanced to human clinical trials for anemia.
Asunto(s)
Anemia/tratamiento farmacológico , Descubrimiento de Drogas , Inhibidores Enzimáticos/farmacología , Prolina Dioxigenasas del Factor Inducible por Hipoxia/antagonistas & inhibidores , Piridazinas/farmacología , Pirimidinas/farmacología , Administración Oral , Anemia/enzimología , Animales , Relación Dosis-Respuesta a Droga , Inhibidores Enzimáticos/administración & dosificación , Inhibidores Enzimáticos/química , Humanos , Prolina Dioxigenasas del Factor Inducible por Hipoxia/metabolismo , Ratones , Ratones Endogámicos C57BL , Modelos Moleculares , Estructura Molecular , Piridazinas/administración & dosificación , Piridazinas/química , Pirimidinas/administración & dosificación , Pirimidinas/química , Ratas , Ratas Sprague-Dawley , Relación Estructura-ActividadRESUMEN
A class of 3,5-diphenyl-1,2,4-oxadiazole based compounds have been identified as potent sphingosine-1-phosphate-1 (S1P1) receptor agonists with minimal affinity for the S1P2 and S1P3 receptor subtypes. Analogue 26 (S1P1 IC50 = 0.6 nM) has an excellent pharmacokinetics profile in the rat and dog and is efficacious in a rat skin transplant model, indicating that S1P3 receptor agonism is not a component of immunosuppressive efficacy.
Asunto(s)
Inmunosupresores/síntesis química , Oxadiazoles/síntesis química , Receptores de Lisoesfingolípidos/agonistas , Animales , Células CHO , Cricetinae , Cricetulus , Perros , Supervivencia de Injerto , Inmunosupresores/farmacocinética , Inmunosupresores/farmacología , Recuento de Linfocitos , Oxadiazoles/farmacocinética , Oxadiazoles/farmacología , Ensayo de Unión Radioligante , Ratas , Ratas Endogámicas Lew , Ratas Sprague-Dawley , Trasplante de Piel , Relación Estructura-ActividadRESUMEN
Hit-to-lead efforts resulted in the discovery of compound 19, a potent CYP11B2 inhibitor that displays high selectivity vs related CYPs, good pharmacokinetic properties in rat and rhesus, and lead-like physical properties. In a rhesus pharmacodynamic model, compound 19 displays robust, dose-dependent aldosterone lowering efficacy, with no apparent effect on cortisol levels.
RESUMEN
We report the discovery of a benzimidazole series of CYP11B2 inhibitors. Hit-to-lead and lead optimization studies identified compounds such as 32, which displays potent CYP11B2 inhibition, high selectivity versus related CYP targets, and good pharmacokinetic properties in rat and rhesus. In a rhesus pharmacodynamic model, 32 produces dose-dependent aldosterone lowering efficacy, with no apparent effect on cortisol levels.
RESUMEN
Moderately potent, selective S1P(1) receptor agonists identified from high-throughput screening have been adapted into lipophilic tails for a class of orally bioavailable amino acid-based S1P(1) agonists represented by 7. Many of the new compounds are potent S1P(1) agonists that select against the S1P(2), S1P(3), and S1P(4) (although not S1P(5)) receptor subtypes. Analogues 18 and 24 are highly orally bioavailable and possess excellent pharmacokinetic profiles in the rat, dog, and rhesus monkey.
Asunto(s)
Azetidinas/farmacología , Inmunosupresores/farmacología , Receptores de Lisoesfingolípidos/agonistas , Administración Oral , Animales , Azetidinas/farmacocinética , Disponibilidad Biológica , Células CHO , Cricetinae , Perros , Diseño de Fármacos , Guanosina 5'-O-(3-Tiotrifosfato)/metabolismo , Humanos , Inmunosupresores/farmacocinética , Linfocitos/efectos de los fármacos , Macaca mulatta , Ratones , Ratas , Ratas Endogámicas Lew , Relación Estructura-ActividadRESUMEN
We report herein the identification of MK-4409, a potent and selective fatty acid amide hydrolase (FAAH) inhibitor. Starting from a high throughput screening (HTS) hit, medicinal chemistry efforts focused on optimizing of FAAH inhibition in vitro potency, improving the pharmacokinetic (PK) profile, and increasing in vivo efficacy in rodent inflammatory and neuropathic pain assays.
RESUMEN
We report herein the discovery of a fatty acid amide hydrolase (FAAH) positron emission tomography (PET) tracer. Starting from a pyrazole lead, medicinal chemistry efforts directed toward reducing lipophilicity led to the synthesis of a series of imidazole analogues. Compound 6 was chosen for further profiling due to its appropriate physical chemical properties and excellent FAAH inhibition potency across species. [(11)C]-6 (MK-3168) exhibited good brain uptake and FAAH-specific signal in rhesus monkeys and is a suitable PET tracer for imaging FAAH in the brain.
RESUMEN
The discovery of 1,3,8-triazaspiro[4.5]decane-2,4-diones (spirohydantoins) as a structural class of pan-inhibitors of the prolyl hydroxylase (PHD) family of enzymes for the treatment of anemia is described. The initial hit class, spirooxindoles, was identified through affinity selection mass spectrometry (AS-MS) and optimized for PHD2 inhibition and optimal PK/PD profile (short-acting PHDi inhibitors). 1,3,8-Triazaspiro[4.5]decane-2,4-diones (spirohydantoins) were optimized as an advanced lead class derived from the original spiroindole hit. A new set of general conditions for C-N coupling, developed using a high-throughput experimentation (HTE) technique, enabled a full SAR analysis of the spirohydantoins. This rapid and directed SAR exploration has resulted in the first reported examples of hydantoin derivatives with good PK in preclinical species. Potassium channel off-target activity (hERG) was successfully eliminated through the systematic introduction of acidic functionality to the molecular structure. Undesired upregulation of alanine aminotransferese (ALT) liver enzymes was mitigated and a robust on-/off-target margin was achieved. Spirohydantoins represent a class of highly efficacious, short-acting PHD1-3 inhibitors causing a robust erythropoietin (EPO) upregulation in vivo in multiple preclinical species. This profile deems spirohydantoins as attractive short-acting PHDi inhibitors with the potential for treatment of anemia.
Asunto(s)
Anemia/tratamiento farmacológico , Compuestos Aza/síntesis química , Hidantoínas/síntesis química , Factor 1 Inducible por Hipoxia/metabolismo , Procolágeno-Prolina Dioxigenasa/antagonistas & inhibidores , Compuestos de Espiro/síntesis química , Animales , Compuestos Aza/farmacocinética , Compuestos Aza/farmacología , Perros , Canal de Potasio ERG1 , Eritropoyetina/biosíntesis , Canales de Potasio Éter-A-Go-Go/metabolismo , Ensayos Analíticos de Alto Rendimiento , Humanos , Hidantoínas/farmacocinética , Hidantoínas/farmacología , Prolina Dioxigenasas del Factor Inducible por Hipoxia , Indoles/síntesis química , Indoles/farmacocinética , Indoles/farmacología , Hígado/efectos de los fármacos , Hígado/enzimología , Macaca mulatta , Espectrometría de Masas , Ratones , Ratones Endogámicos C57BL , Unión Proteica , Ratas , Compuestos de Espiro/farmacocinética , Compuestos de Espiro/farmacología , Relación Estructura-Actividad , Regulación hacia ArribaRESUMEN
Structure-activity relationship (SAR) studies of 3-arylpropionic acids-a class of novel S1P(1) selective agonists-by introducing substitution to the propionic acid chain and replacing the adjacent phenyl ring with pyridine led to a series of modified 3-arylpropionic acids with enhanced half-life in rat. These analogs (e.g., cyclopropanecarboxylic acids) exhibited longer half-life in rat than did unmodified 3-arylpropionic acids. This result suggests that metabolic oxidation at the propionic acid chain, particularly at the C3 benzylic position of 3-arylpropionic acids, is probably responsible for their short half-life in rodent.
Asunto(s)
Propionatos/síntesis química , Propionatos/farmacología , Receptores de Lisoesfingolípidos/agonistas , Animales , Disponibilidad Biológica , Células CHO , Cricetinae , Cricetulus , Guanosina 5'-O-(3-Tiotrifosfato)/metabolismo , Semivida , Recuento de Linfocitos , Linfocitos/efectos de los fármacos , Linfocitos/metabolismo , Masculino , Ratones , Ratas , Ratas Sprague-Dawley , Relación Estructura-ActividadRESUMEN
A series of 2-aryl(pyrrolidin-4-yl)acetic acids were synthesized and their biological activities were evaluated as agonists of S1P receptors. These analogs were able to induce lowering of lymphocyte counts in the peripheral blood of mice and were found to have good overall pharmacokinetic properties in rat.
Asunto(s)
Acetatos/farmacología , Pirrolidinas/farmacología , Receptores de Lisoesfingolípidos/agonistas , Acetatos/síntesis química , Acetatos/química , Animales , Recuento de Linfocitos , Lisofosfolípidos/antagonistas & inhibidores , Ratones , Estructura Molecular , Unión Proteica/efectos de los fármacos , Pirrolidinas/síntesis química , Pirrolidinas/química , Ratas , Esfingosina/análogos & derivados , Esfingosina/antagonistas & inhibidores , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
Novel series of sphingosine-1-phosphate (S1P) receptor agonists were developed through a systematic SAR aimed to achieve high selectivity for a single member of the S1P family of receptors, S1P1. The optimized structure represents a highly S1P1-selective and efficacious agonist: S1P1/S1P2, S1P1/S1P3, S1P1/S1P4>10,000-fold, S1P1/S1P5>600-fold, while EC50 (S1P1) <0.2 nM. In vivo experiments are consistent with S1P1 receptor agonism alone being sufficient for achieving desired lymphocyte-lowering effect.
Asunto(s)
Inmunosupresores/síntesis química , Inmunosupresores/farmacología , Linfocitos/efectos de los fármacos , Receptores de Lisoesfingolípidos/agonistas , Animales , Células CHO , Cricetinae , Recuento de Linfocitos , Linfocitos/citología , Relación Estructura-ActividadRESUMEN
A series of 2,5-cis-disubstituted pyrrolidines were synthesized and evaluated as S1P receptor agonists. Compounds 15-21 were identified with good selectivity over S1P3 which lowered circulating lymphocytes after oral administration in mice.
Asunto(s)
Ácidos Carboxílicos/química , Ácidos Carboxílicos/farmacología , Pirrolidinas/química , Receptores de Lisoesfingolípidos/agonistas , Administración Oral , Animales , Ácidos Carboxílicos/síntesis química , Estructura Molecular , Ratas , Receptores de Lisoesfingolípidos/metabolismo , Relación Estructura-ActividadRESUMEN
A series of 3-arylpropionic acids were synthesized as S1P1 receptor agonists. Structure-activity relationship studies on the pendant phenyl ring revealed several structural features offering selectivity of S1P1 binding against S1P2-5. These highly selective S1P1 agonists induced peripheral blood lymphocyte lowering in mice and one of them was found to be efficacious in a rat skin transplantation model, supporting that S1P1 agonism is primarily responsible for the immunosuppressive efficacy observed in preclinical animal models.
Asunto(s)
Inmunosupresores/farmacología , Fenilpropionatos/síntesis química , Fenilpropionatos/farmacología , Receptores de Lisoesfingolípidos/agonistas , Animales , Células CHO , Cricetinae , Ligandos , Recuento de Linfocitos/veterinaria , Ratones , Ratas , Trasplante de Piel , Relación Estructura-ActividadRESUMEN
The novel immunosuppressive agent FTY720 (1) is phosphorylated in vivo in a variety of species yielding an active metabolite that is an agonist of four of the five known G-protein-coupled sphingosine-1-phosphate (S1P) receptors. A synthesis amenable to producing gram quantities of the stereoisomeric phosphate esters, a determination of their absolute stereochemistry via an enantioselective synthesis and their characterization as S1P receptor agonists and antagonists is reported.
Asunto(s)
Inmunosupresores/síntesis química , Organofosfatos/síntesis química , Glicoles de Propileno/síntesis química , Animales , Células CHO , Cricetinae , Clorhidrato de Fingolimod , Humanos , Conformación Molecular , Esfingosina/análogos & derivadosRESUMEN
Sphingosine-1-phosphate (S1P), a lipid signaling molecule that regulates many cellular functions, is synthesized from sphingosine and ATP by the action of sphingosine kinase. Two such kinases have been identified, SPHK1 and SPHK2. To begin to investigate the physiological functions of sphingosine kinase and S1P signaling, we generated mice deficient in SPHK1. Sphk1 null mice were viable, fertile, and without any obvious abnormalities. Total SPHK activity in most Sphk1-/-tissues was substantially, but not completely, reduced indicating the presence of multiple sphingosine kinases. S1P levels in most tissues from the Sphk1-/- mice were not markedly decreased. In serum, however, there was a significant decrease in the S1P level. Although S1P signaling regulates lymphocyte trafficking, lymphocyte distribution was unaffected in lymphoid organs of Sphk1-/- mice. The immunosuppressant FTY720 was phosphorylated and elicited lymphopenia in the Sphk1 null mice showing that SPHK1 is not required for the functional activation of this sphingosine analogue prodrug. The results with these Sphk1 null mice reveal that some key physiologic processes that require S1P receptor signaling, such as vascular development and proper lymphocyte distribution, can occur in the absence of SPHK1.
Asunto(s)
Linfopenia/etiología , Fosfotransferasas (Aceptor de Grupo Alcohol)/deficiencia , Glicoles de Propileno/toxicidad , Esfingosina/análogos & derivados , Animales , Secuencia de Bases , ADN/genética , Clorhidrato de Fingolimod , Inmunosupresores/metabolismo , Inmunosupresores/toxicidad , Linfocitos/efectos de los fármacos , Linfocitos/fisiología , Linfopenia/inducido químicamente , Linfopenia/enzimología , Lisofosfolípidos/metabolismo , Ratones , Ratones Noqueados , Fosforilación , Fosfotransferasas (Aceptor de Grupo Alcohol)/genética , Fosfotransferasas (Aceptor de Grupo Alcohol)/fisiología , Profármacos/metabolismo , Profármacos/toxicidad , Glicoles de Propileno/metabolismo , Receptores de Lisoesfingolípidos/metabolismo , Transducción de Señal , Esfingosina/metabolismoRESUMEN
3-(N-Alkyl)aminopropylphosphonic acids are potent agonists of four of the five known sphingosine-1-phosphate (S1P) receptor subtypes.
Asunto(s)
Receptores Acoplados a Proteínas G/agonistas , Receptores de Lisoesfingolípidos/agonistas , Animales , Células CHO , Cricetinae , Humanos , Concentración 50 Inhibidora , Ligandos , Organofosfonatos/síntesis química , Compuestos Organofosforados/síntesis química , Radioisótopos de Fósforo , Relación Estructura-ActividadRESUMEN
Blood lymphocyte numbers, essential for the development of efficient immune responses, are maintained by recirculation through secondary lymphoid organs. We show that lymphocyte trafficking is altered by the lysophospholipid sphingosine-1-phosphate (S1P) and by a phosphoryl metabolite of the immunosuppressive agent FTY720. Both species were high-affinity agonists of at least four of the five S1P receptors. These agonists produce lymphopenia in blood and thoracic duct lymph by sequestration of lymphocytes in lymph nodes, but not spleen. S1P receptor agonists induced emptying of lymphoid sinuses by retention of lymphocytes on the abluminal side of sinus-lining endothelium and inhibition of egress into lymph. Inhibition of lymphocyte recirculation by activation of S1P receptors may result in therapeutically useful immunosuppression.
Asunto(s)
Linfocitos B/fisiología , Lisofosfolípidos , Organofosfatos/farmacología , Organofosfonatos/farmacología , Glicoles de Propileno/metabolismo , Receptores de Superficie Celular/agonistas , Receptores Acoplados a Proteínas G , Esfingosina/análogos & derivados , Esfingosina/farmacología , Linfocitos T/fisiología , Animales , Linfocitos B/efectos de los fármacos , Unión Competitiva , Células CHO , Calcio/metabolismo , Cricetinae , AMP Cíclico/metabolismo , Clorhidrato de Fingolimod , Guanosina 5'-O-(3-Tiotrifosfato)/metabolismo , Humanos , Inmunosupresores/metabolismo , Inmunosupresores/farmacología , Ligandos , Ganglios Linfáticos/citología , Ganglios Linfáticos/efectos de los fármacos , Recuento de Linfocitos , Linfopenia/inducido químicamente , Ratones , Organofosfatos/síntesis química , Organofosfatos/química , Organofosfatos/metabolismo , Organofosfonatos/síntesis química , Organofosfonatos/química , Organofosfonatos/metabolismo , Fosforilación , Glicoles de Propileno/farmacología , Ratas , Receptores de Superficie Celular/metabolismo , Receptores Lisofosfolípidos , Esfingosina/metabolismo , Bazo/citología , Bazo/efectos de los fármacos , Estereoisomerismo , Linfocitos T/efectos de los fármacosRESUMEN
Alteration in lymphocyte trafficking and prevention of graft rejection in rodents observed on exposure to FTY720 (1) or its corresponding phosphate ester 2 can be induced by the systemic administration of potent sphingosine-1-phosphate receptor agonists exemplified by 19. The similar S1P receptor profiles of 2 and 19 coupled with their comparable potency in vivo supports a connection between S1P receptor agonism and immunosuppressive efficacy.
Asunto(s)
Factores Inmunológicos/administración & dosificación , Factores Inmunológicos/química , Glicoles de Propileno/administración & dosificación , Glicoles de Propileno/química , Receptores de Lisoesfingolípidos/agonistas , Animales , Células CHO , Cricetinae , Clorhidrato de Fingolimod , Humanos , Factores Inmunológicos/metabolismo , Inmunosupresores/administración & dosificación , Inmunosupresores/química , Inmunosupresores/metabolismo , Inyecciones Intravenosas , Masculino , Ratones , Glicoles de Propileno/metabolismo , Ratas , Ratas Sprague-Dawley , Receptores de Lisoesfingolípidos/metabolismo , Esfingosina/análogos & derivadosRESUMEN
A series of conformationally constrained 3-(N-alkylamino)propylphosphonic acids were systematically synthesized and their activities as S1P receptor agonists were evaluated. Several pyrrolidine and cyclohexane analogs had S1P receptor profiles comparable to the acyclic lead compound, 3-(N-tetradecylamino)propylphosphonic acid (3), lowered circulating lymphocytes in mice after iv administration and were thus identified as being suitable for further investigations.