RESUMEN
A series of beta-aminoamides bearing triazolopiperazines have been discovered as potent, selective, and orally active dipeptidyl peptidase IV (DPP-4) inhibitors by extensive structure-activity relationship (SAR) studies around the triazolopiperazine moiety. Among these, compound 34b with excellent in vitro potency (IC50 = 4.3 nM) against DPP-4, high selectivity over other enzymes, and good pharmacokinetic profiles exhibited pronounced in vivo efficacy in an oral glucose tolerance test (OGTT) in lean mice. On the basis of these properties, compound 34b has been profiled in detail. Further refinement of the triazolopiperazines resulted in the discovery of a series of extremely potent compounds with subnanomolar activity against DPP-4 (42b- 49b), that is, 4-fluorobenzyl-substituted compound 46b, which is notable for its superior potency (IC50 = 0.18 nM). X-ray crystal structure determination of compounds 34b and 46b in complex with DPP-4 enzyme revealed that (R)-stereochemistry at the 8-position of triazolopiperazines is strongly preferred over (S) with respect to DPP-4 inhibition.
Asunto(s)
Amidas/síntesis química , Inhibidores de la Dipeptidil-Peptidasa IV , Piperazinas/síntesis química , Pirazinas/síntesis química , Triazoles/síntesis química , Amidas/farmacocinética , Amidas/farmacología , Animales , Cristalografía por Rayos X , Dipeptidil Peptidasa 4/química , Perros , Prueba de Tolerancia a la Glucosa , Haplorrinos , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , Piperazinas/farmacocinética , Piperazinas/farmacología , Pirazinas/farmacocinética , Pirazinas/farmacología , Ratas , Estereoisomerismo , Relación Estructura-Actividad , Triazoles/farmacocinética , Triazoles/farmacologíaRESUMEN
Probing with tool molecules, and by modeling and X-ray crystallography the binding modes of two structurally distinct series of DPP-4 inhibitors led to the discovery of a rare aromatic fluorine H-bond and the spatial requirement for better biaryl binding in the DPP-4 enzyme active site. These newly found binding elements were successfully incorporated into novel DPP-4 inhibitors.
Asunto(s)
Dipeptidil Peptidasa 4/química , Sitios de Unión , Química Farmacéutica/métodos , Cristalografía por Rayos X/métodos , Inhibidores de la Dipeptidil-Peptidasa IV , Diseño de Fármacos , Flúor/química , Glicina/química , Humanos , Enlace de Hidrógeno , Concentración 50 Inhibidora , Modelos Químicos , Modelos Teóricos , Conformación Molecular , Programas Informáticos , beta-Alanina/químicaRESUMEN
The synthesis, selectivity, rat pharmacokinetic profile, and drug metabolism profiles of a series of potent fluoroolefin-derived DPP-4 inhibitors (4) are reported. A radiolabeled fluoroolefin 33 was shown to possess a high propensity to form reactive metabolites, thus revealing a potential liability for this class of DPP-4 inhibitors.
Asunto(s)
Alquenos/farmacocinética , Amidas/química , Inhibidores de la Dipeptidil-Peptidasa IV/farmacocinética , Hidrocarburos Fluorados/farmacocinética , Hipoglucemiantes/farmacocinética , Microsomas Hepáticos/efectos de los fármacos , Alquenos/síntesis química , Animales , Inhibidores de la Dipeptidil-Peptidasa IV/síntesis química , Hidrocarburos Fluorados/síntesis química , Hipoglucemiantes/síntesis química , Microsomas Hepáticos/patología , Modelos Químicos , Imitación Molecular , Ratas , Estereoisomerismo , Relación Estructura-Actividad , Especificidad por SustratoRESUMEN
Dipeptidyl peptidase (DPP)-IV inhibitors are a new approach to the treatment of type 2 diabetes. DPP-IV is a member of a family of serine peptidases that includes quiescent cell proline dipeptidase (QPP), DPP8, and DPP9; DPP-IV is a key regulator of incretin hormones, but the functions of other family members are unknown. To determine the importance of selective DPP-IV inhibition for the treatment of diabetes, we tested selective inhibitors of DPP-IV, DPP8/DPP9, or QPP in 2-week rat toxicity studies and in acute dog tolerability studies. In rats, the DPP8/9 inhibitor produced alopecia, thrombocytopenia, reticulocytopenia, enlarged spleen, multiorgan histopathological changes, and mortality. In dogs, the DPP8/9 inhibitor produced gastrointestinal toxicity. The QPP inhibitor produced reticulocytopenia in rats only, and no toxicities were noted in either species for the selective DPP-IV inhibitor. The DPP8/9 inhibitor was also shown to attenuate T-cell activation in human in vitro models; a selective DPP-IV inhibitor was inactive in these assays. Moreover, we found DPP-IV inhibitors that were previously reported to be active in models of immune function to be more potent inhibitors of DPP8/9. These results suggest that assessment of selectivity of potential clinical candidates may be important to an optimal safety profile for this new class of antihyperglycemic agents.
Asunto(s)
Diabetes Mellitus Tipo 2/tratamiento farmacológico , Dipeptidasas/antagonistas & inhibidores , Dipeptidil Peptidasa 4 , Dipeptidil-Peptidasas y Tripeptidil-Peptidasas/antagonistas & inhibidores , Hipoglucemiantes , Inhibidores de Proteasas/uso terapéutico , Animales , Dipeptidil Peptidasa 4/genética , Dipeptidil Peptidasa 4/fisiología , Perros , Femenino , Humanos , Hipoglucemiantes/uso terapéutico , Hipoglucemiantes/toxicidad , Isoleucina/análogos & derivados , Isoleucina/química , Isoleucina/uso terapéutico , Isoleucina/toxicidad , Isomerismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Inhibidores de Proteasas/toxicidad , Ratas , Proteínas Recombinantes/antagonistas & inhibidores , Tiazoles/química , Tiazoles/uso terapéutico , Tiazoles/toxicidadRESUMEN
A series of beta-substituted biarylphenylalanine amides were synthesized and evaluated as inhibitors of dipeptidyl peptidase IV (DPP-4) for the treatment of type 2 diabetes. Optimization of the metabolic profile of early analogues led to the discovery of (2S,3S)-3-amino-4-(3,3-difluoropyrrolidin-1-yl)-N,N-dimethyl-4-oxo-2-(4-[1,2,4]triazolo[1,5-a]pyridin-6-ylphenyl)butanamide (6), a potent, orally active DPP-4 inhibitor (IC(50) = 6.3 nM) with excellent selectivity, oral bioavailability in preclinical species, and in vivo efficacy in animal models. Compound 6 was selected for further characterization as a potential new treatment for type 2 diabetes.
Asunto(s)
Diabetes Mellitus Tipo 2/tratamiento farmacológico , Dipeptidil Peptidasa 4/metabolismo , Hipoglucemiantes/síntesis química , Fenilalanina/análogos & derivados , Inhibidores de Proteasas/síntesis química , Triazoles/síntesis química , Administración Oral , Animales , Disponibilidad Biológica , Canales de Calcio Tipo L/efectos de los fármacos , Línea Celular , Cristalografía por Rayos X , Prueba de Tolerancia a la Glucosa , Humanos , Hipoglucemiantes/química , Hipoglucemiantes/farmacología , Técnicas In Vitro , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Obesos , Microsomas Hepáticos/efectos de los fármacos , Microsomas Hepáticos/metabolismo , Modelos Moleculares , Proteínas Musculares/antagonistas & inhibidores , Músculo Esquelético/metabolismo , Canal de Sodio Activado por Voltaje NAV1.5 , Fenilalanina/síntesis química , Fenilalanina/química , Fenilalanina/farmacología , Inhibidores de Proteasas/química , Inhibidores de Proteasas/farmacología , Conejos , Canales de Sodio , Estereoisomerismo , Relación Estructura-Actividad , Triazoles/química , Triazoles/farmacologíaRESUMEN
The presence of DPPII (dipeptidyl peptidase II; E.C. 3.4.14.2) has been demonstrated in various mammalian tissues. However, a profound molecular and catalytic characterization, including substrate selectivity, kinetics and pH-dependence, has not been conducted. In the present study, DPPII was purified from human seminal plasma to apparent homogeneity with a high yield (40%) purification scheme, including an inhibitor-based affinity chromatographic step. The inhibitor lysyl-piperidide (K(i) approximately 0.9 microM at pH 5.5) was chosen, as it provided a favourable affinity/recovery ratio. The human enzyme appeared as a 120 kDa homodimer. Mass spectrometric analysis after tryptic digestion together with a kinetic comparison indicate strongly its identity with QPP (quiescent cell proline dipeptidase), also called dipeptidyl peptidase 7. pH profiles of both kcat and kcat/K(m) clearly demonstrated that DPPII/QPP possesses an acidic and not a neutral optimum as was reported for QPP. Kinetic parameters of the human natural DPPII for dipeptide-derived chromogenic [pNA (p-nitroanilide)] and fluorogenic [4Me2NA (4-methoxy-2-naphthylamide)] substrates were determined under different assay conditions. DPPII preferred the chromogenic pNA-derived substrates over the fluorogenic 4Me2NA-derived substrates. Natural human DPPII showed high efficiency towards synthetic substrates containing proline at the P1 position and lysine at P2. The importance of the P1' group for P2 and P1 selectivity was revealed, explaining many discrepancies in the literature. Furthermore, substrate preferences of human DPPII and dipeptidyl peptidase IV were compared based on their selectivity constants (kcat/K(m)). Lys-Pro-pNA (k(cat)/K(m) 4.1x10(6) s(-1) x M(-1)) and Ala-Pro-pNA (kcat/K(m) 2.6x10(6) s(-1) x M(-1)) were found to be the most sensitive chromogenic substrates for human DPPII, but were less selective than Lys-Ala-pNA (kcat/K(m) 0.4x10(6) s(-1) x M(-1)).
Asunto(s)
Dipeptidasas/genética , Dipéptidos/metabolismo , Dipeptidil-Peptidasas y Tripeptidil-Peptidasas/metabolismo , Secuencia de Aminoácidos , Animales , Dipeptidil Peptidasa 4/metabolismo , Dipeptidil-Peptidasas y Tripeptidil-Peptidasas/química , Inhibidores Enzimáticos/química , Estabilidad de Enzimas , Humanos , Concentración de Iones de Hidrógeno , Hidrólisis , Cinética , Ratones , Datos de Secuencia Molecular , Estructura Molecular , Peso Molecular , Nanotecnología/métodos , Ratas , Proteínas Recombinantes/química , Alineación de Secuencia/métodos , Espectrometría de Masa por Ionización de Electrospray/métodos , Especificidad por SustratoRESUMEN
A novel series of beta-amino amides incorporating fused heterocycles, i.e., triazolopiperazines, were synthesized and evaluated as inhibitors of dipeptidyl peptidase IV (DPP-IV) for the treatment of type 2 diabetes. (2R)-4-Oxo-4-[3-(trifluoromethyl)-5,6-dihydro[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl]-1-(2,4,5-trifluorophenyl)butan-2-amine (1) is a potent, orally active DPP-IV inhibitor (IC(50) = 18 nM) with excellent selectivity over other proline-selective peptidases, oral bioavailability in preclinical species, and in vivo efficacy in animal models. MK-0431, the phosphate salt of compound 1, was selected for development as a potential new treatment for type 2 diabetes.
Asunto(s)
Diabetes Mellitus Tipo 2/tratamiento farmacológico , Dipeptidil Peptidasa 4/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Hipoglucemiantes/farmacología , Pirazinas/química , Pirazinas/farmacología , Triazoles/química , Triazoles/farmacología , Administración Oral , Animales , Sitios de Unión , Bioquímica/métodos , Glucemia/análisis , Cristalografía por Rayos X , Dipeptidil Peptidasa 4/química , Dipeptidil Peptidasa 4/metabolismo , Perros , Relación Dosis-Respuesta a Droga , Evaluación Preclínica de Medicamentos/métodos , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacocinética , Glucagón/sangre , Glucagón/efectos de los fármacos , Péptido 1 Similar al Glucagón , Prueba de Tolerancia a la Glucosa , Hipoglucemiantes/química , Hipoglucemiantes/farmacocinética , Ratones , Ratones Endogámicos C57BL , Modelos Moleculares , Fragmentos de Péptidos/sangre , Fragmentos de Péptidos/efectos de los fármacos , Conformación Proteica , Precursores de Proteínas/sangre , Precursores de Proteínas/efectos de los fármacos , Pirazinas/farmacocinética , Ratas , Fosfato de Sitagliptina , Relación Estructura-Actividad , Triazoles/farmacocinéticaRESUMEN
A series of substituted imidazopiperidine amides has been prepared and evaluated for inhibition of dipeptidyl peptidase IV (DPP-4). Substitution at the 1- and 3-positions produced increased selectivity for DPP-4 relative to DPP-8 and DPP-9. Compounds in this series had IC(50) values as low as 5.8 nM for inhibition of DPP-4.
Asunto(s)
Diabetes Mellitus/tratamiento farmacológico , Inhibidores de la Dipeptidil-Peptidasa IV , Hipoglucemiantes/farmacología , Piperidinas/farmacología , Inhibidores de Proteasas/farmacología , Amidas/química , Humanos , Hipoglucemiantes/uso terapéutico , Piperidinas/química , Piperidinas/uso terapéutico , Inhibidores de Proteasas/química , Inhibidores de Proteasas/uso terapéuticoRESUMEN
Various beta-amino amides containing triazolopiperazine heterocycles have been prepared and evaluated as potent, selective, orally active dipeptidyl peptidase IV (DPP-4) inhibitors. These compounds display excellent oral bioavailability and good overall pharmacokinetic profiles in preclinical species. Moreover, in vivo efficacy in an oral glucose tolerance test in lean mice is demonstrated.
Asunto(s)
Amidas/química , Amidas/farmacología , Inhibidores de la Dipeptidil-Peptidasa IV , Piperazinas/química , Inhibidores de Proteasas/química , Inhibidores de Proteasas/farmacología , Amidas/síntesis química , Amidas/farmacocinética , Animales , Cristalografía por Rayos X , Diseño de Fármacos , Evaluación Preclínica de Medicamentos , Modelos Moleculares , Inhibidores de Proteasas/síntesis química , Inhibidores de Proteasas/farmacocinética , RatasRESUMEN
Substituted 3-aminopiperidines 3 were evaluated as DPP-4 inhibitors. The inhibitors showed good DPP-4 potency with superb selectivity over other peptidases (QPP, DPP8, and DPP9). Selected DPP-4 inhibitors were further evaluated for their hERG potassium channel, calcium channel, Cyp2D6, and pharmacokinetic profiles.
Asunto(s)
Inhibidores de la Dipeptidil-Peptidasa IV , Piperidinas/química , Piperidinas/farmacología , Animales , Área Bajo la Curva , Disponibilidad Biológica , Semivida , Humanos , Hipoglucemiantes/química , Hipoglucemiantes/farmacología , Estructura Molecular , Piperidinas/sangre , Ratas , Relación Estructura-ActividadRESUMEN
A novel series of 4-arylcyclohexylalanine DPP-4 inhibitors was synthesized and tested for inhibitory activity as well as selectivity over the related proline-specific enzymes DPP-8 and DPP-9. Optimization of this series led to 28 (DPP-4 IC(50)=4.8 nM), which showed an excellent pharmacokinetic profile across several preclinical species. Evaluation of 28 in an oral glucose tolerance test demonstrated that this compound effectively reduced glucose excursion in lean mice.
Asunto(s)
Alanina/análogos & derivados , Inhibidores de la Dipeptidil-Peptidasa IV , Inhibidores de Proteasas/farmacología , Administración Oral , Alanina/química , Alanina/farmacología , Animales , Área Bajo la Curva , Ratones , Modelos Moleculares , Inhibidores de Proteasas/administración & dosificación , Inhibidores de Proteasas/química , Inhibidores de Proteasas/farmacocinéticaRESUMEN
A novel series of 4-aminophenylalanine and 4-aminocyclohexylalanine derivatives were designed and evaluated as inhibitors of dipeptidyl peptidase IV (DPP-4). The phenylalanine series afforded compounds such as 10 that were potent and selective (DPP-4, IC(50)=28nM), but exhibited limited oral bioavailability. The corresponding cyclohexylalanine derivatives such as 25 afforded improved PK exposure and efficacy in a murine OGTT experiment. The X-ray crystal structure of 25 bound to the DPP-4 active site is presented.
Asunto(s)
Alanina/análogos & derivados , Ciclohexanos/farmacología , Inhibidores de la Dipeptidil-Peptidasa IV , Inhibidores Enzimáticos/farmacología , Fenilalanina/análogos & derivados , Administración Oral , Alanina/química , Alanina/farmacología , Animales , Sitios de Unión , Disponibilidad Biológica , Cristalografía por Rayos X , Ciclohexanos/química , Dipeptidil Peptidasa 4/química , Inhibidores Enzimáticos/química , Prueba de Tolerancia a la Glucosa , Ratones , Modelos Moleculares , Estructura Molecular , Fenilalanina/química , Fenilalanina/farmacología , Relación Estructura-ActividadRESUMEN
Following the discovery of N-acyl-1,4-diazepan-2-one as a novel pharmacophore for potent and selective DPP-4 inhibitors, optimization of this new lead with different substitution on the seven-membered ring resulted in several highly potent and selective, orally bioavailable, and efficacious DPP-4 inhibitors, such as 3R-methyl-1-cyclopropyl-1,4-diazepan-2-one derivative 9i (DPP-4 IC(50)=8.0 nM) and 3R,6R-dimethyl-1,4-diazepan-2-one derivative 14a (DPP-4 IC(50)=9.7 nM).
Asunto(s)
Azepinas/síntesis química , Química Farmacéutica/métodos , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Inhibidores de la Dipeptidil-Peptidasa IV , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/farmacología , Administración Oral , Animales , Azepinas/farmacología , Diseño de Fármacos , Concentración 50 Inhibidora , Masculino , Ratones , Modelos Químicos , Conformación Molecular , Ratas , Ratas Sprague-DawleyRESUMEN
A series of beta-aminoamides bearing triazolopiperazines has been prepared and evaluated as potent, selective, orally active dipeptidyl peptidase IV (DPP-4) inhibitors. Efforts at optimization of the beta-aminoamide series, which ultimately led to the discovery of JANUVIA (sitagliptin phosphate, compound 1), are described.
Asunto(s)
Amidas/química , Inhibidores de la Dipeptidil-Peptidasa IV , Piperazinas/química , Inhibidores de Proteasas/farmacología , Pirazinas/farmacología , Triazoles/química , Animales , Inhibidores de Proteasas/síntesis química , Pirazinas/química , Ratas , Fosfato de Sitagliptina , Relación Estructura-Actividad , Triazoles/farmacologíaRESUMEN
Replacement of the triazolopiperazine ring of sitagliptin (DPP-4 IC(50)=18nM) with 3-(2,2,2-trifluoroethyl)-1,4-diazepan-2-one gave dipeptidyl peptidase IV (DPP-4) inhibitor 1 which is potent (DPP-4 IC(50)=2.6nM), selective, and efficacious in an oral glucose tolerance test in mice. It was selected for extensive preclinical development as a potential back-up candidate to sitagliptin.
Asunto(s)
Azepinas/química , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Inhibidores de la Dipeptidil-Peptidasa IV , Hipoglucemiantes/química , Inhibidores de Proteasas/química , Animales , Azepinas/uso terapéutico , Cristalografía por Rayos X , Dipeptidil Peptidasa 4/química , Hipoglucemiantes/uso terapéutico , Inhibidores de Proteasas/uso terapéutico , Conformación Proteica , Pirazinas/química , Pirazinas/uso terapéutico , Ratas , Ratas Endogámicas , Fosfato de Sitagliptina , Triazoles/química , Triazoles/uso terapéuticoRESUMEN
anti-Substituted beta-methylphenylalanine derived amides have been shown to be potent DPP-IV inhibitors exhibiting excellent selectivity over both DPP8 and DPP9. The optimized compound exhibited good pharmacokinetic profiles in three preclinical species.
Asunto(s)
Inhibidores de la Adenosina Desaminasa , Glicoproteínas/antagonistas & inhibidores , Inhibidores de Proteasas/administración & dosificación , Inhibidores de Proteasas/farmacología , Piridonas/administración & dosificación , Piridonas/farmacología , Adenosina Desaminasa/metabolismo , Administración Oral , Amidas/química , Animales , Disponibilidad Biológica , Dipeptidil Peptidasa 4/metabolismo , Perros , Glicoproteínas/metabolismo , Humanos , Concentración 50 Inhibidora , Macaca mulatta , Estructura Molecular , Nitrógeno/química , Inhibidores de Proteasas/química , Inhibidores de Proteasas/farmacocinética , Piridonas/química , Piridonas/farmacocinética , Ratas , Sensibilidad y Especificidad , Relación Estructura-Actividad , Especificidad por SustratoRESUMEN
A novel series of oxadiazole based amides have been shown to be potent DPP-4 inhibitors. The optimized compound 43 exhibited excellent selectivity over a variety of DPP-4 homologs.
Asunto(s)
Inhibidores de la Dipeptidil-Peptidasa IV , Oxadiazoles/administración & dosificación , Oxadiazoles/farmacología , Inhibidores de Proteasas/administración & dosificación , Inhibidores de Proteasas/farmacología , Administración Oral , Animales , Sitios de Unión , Disponibilidad Biológica , Perros , Activación Enzimática/efectos de los fármacos , Canales de Potasio Éter-A-Go-Go/efectos de los fármacos , Humanos , Modelos Moleculares , Conformación Molecular , Oxadiazoles/química , Inhibidores de Proteasas/química , Conformación Proteica , Ratas , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
anti-Substituted beta-methylphenylalanine derived amides have been shown to be potent DPP-IV inhibitors exhibiting excellent selectivity over both DPP8 and DPP9. These are among the most potent compounds reported to date lacking an electrophilic trap. The most potent compound among these is 5-oxo-1,2,4-oxadiazole 44, which is a 3 nM DPP-IV inhibitor.
Asunto(s)
Dipeptidil Peptidasa 4/efectos de los fármacos , Fenilalanina/farmacología , Inhibidores de Proteasas/farmacología , Fenilalanina/química , Inhibidores de Proteasas/químicaRESUMEN
A series of beta-aminoacylpiperidines bearing various fused five-membered heterocyclic rings was synthesized as dipeptidyl peptidase IV inhibitors. Potent and relatively selective inhibition could be obtained, depending on choice of heterocycle, regioisomerism, and substitution. In particular, one analog (74, DPP-IV IC50=26 nM) exhibited good oral bioavailability and acceptable half-life in the rat, albeit with rather high clearance.
Asunto(s)
Dipeptidil Peptidasa 4/metabolismo , Piperidinas/síntesis química , Piperidinas/farmacología , Inhibidores de Proteasas/síntesis química , Inhibidores de Proteasas/farmacología , Isoxazoles , Oxazoles , Piperidinas/química , Inhibidores de Proteasas/química , Pirazoles , TiazolesRESUMEN
anti-Substituted biaryl beta-methylphenylalanine derived amides have been shown to be potent DPP-IV inhibitors that suffer from suboptimal selectivity and pharmacokinetics. This letter describes the substitution of the beta-methyl substituent with beta-polar substituents, culminating in the discovery of a beta-dimethylamide substituted phenylalanine derivative with an excellent potency, selectivity, and pharmacokinetic profile.