RESUMEN
Fatty acid amide hydrolase (FAAH), an amidase-signature family member, is an integral membrane enzyme that degrades lipid amides including the endogenous cannabinoid anandamide and the sleep-inducing molecule oleamide. Both genetic knock out and pharmacological administration of FAAH inhibitors in rodent models result in analgesic, anxiolytic, and antiinflammatory phenotypes. Targeting FAAH activity, therefore, presents a promising new therapeutic strategy for the treatment of pain and other neurological-related or inflammatory disorders. Nearly all FAAH inhibitors known to date attain their binding potency through a reversible or irreversible covalent modification of the nucleophile Ser241 in the unusual Ser-Ser-Lys catalytic triad. Here, we report the discovery and mechanism of action of a series of ketobenzimidazoles as unique and potent noncovalent FAAH inhibitors. Compound 2, a representative of these ketobenzimidazoles, was designed from a series of ureas that were identified from high-throughput screening. While urea compound 1 is characterized as an irreversible covalent inhibitor, the cocrystal structure of FAAH complexed with compound 2 reveals that these ketobenzimidazoles, though containing a carbonyl moiety, do not covalently modify Ser241. These inhibitors achieve potent inhibition of FAAH activity primarily from shape complementarity to the active site and through numerous hydrophobic interactions. These noncovalent compounds exhibit excellent selectivity and good pharmacokinetic properties. The discovery of this distinctive class of inhibitors opens a new avenue for modulating FAAH activity through nonmechanism-based inhibition.
Asunto(s)
Amidohidrolasas/antagonistas & inhibidores , Bencimidazoles/aislamiento & purificación , Bencimidazoles/metabolismo , Descubrimiento de Drogas/métodos , Inhibidores Enzimáticos/aislamiento & purificación , Inhibidores Enzimáticos/metabolismo , Modelos Moleculares , Animales , Bencimidazoles/farmacocinética , Cumarinas , Cristalización , Inhibidores Enzimáticos/farmacocinética , Escherichia coli , Humanos , Estructura Molecular , Ratas , Espectrofotometría Ultravioleta , Espectrometría de Masas en Tándem , Urea/metabolismoRESUMEN
Sphingosine-1-phosphate (S1P) signaling plays a vital role in mitogenesis, cell migration and angiogenesis. Sphingosine kinases (SphKs) catalyze a key step in sphingomyelin metabolism that leads to the production of S1P. There are two isoforms of SphK and observations made with SphK deficient mice show the two isoforms can compensate for each other's loss. Thus, inhibition of both isoforms is likely required to block SphK dependent angiogenesis. A structure based approach was used to design and synthesize a series of SphK inhibitors resulting in the identification of the first potent inhibitors of both isoforms of human SphK. Additionally, to our knowledge, this series of inhibitors contains the only sufficiently potent inhibitors of murine SphK1 with suitable physico-chemical properties to pharmacologically interrogate the role of SphK1 in rodent models and to reproduce the phenotype of SphK1 (-/-) mice.
Asunto(s)
Diseño de Fármacos , Inhibidores Enzimáticos/síntesis química , Fosfotransferasas (Aceptor de Grupo Alcohol)/antagonistas & inhibidores , Fosfotransferasas (Aceptor de Grupo Alcohol)/química , Bibliotecas de Moléculas Pequeñas/síntesis química , Animales , Células Cultivadas , Cristalografía por Rayos X , Activación Enzimática/efectos de los fármacos , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Humanos , Ratones , Estructura Molecular , Isoformas de Proteínas/química , Ratas , Bibliotecas de Moléculas Pequeñas/química , Bibliotecas de Moléculas Pequeñas/farmacología , Relación Estructura-ActividadRESUMEN
The optimization of a series of 8-aza-quinazolinone analogs for antagonist activity against the CXCR3 receptor is reported. Compounds were optimized to avoid the formation of active metabolites and time-dependent-inhibitors of CYP3A4. In addition, antagonists showed potent against CXCR3 activity in whole blood and optimized to avoid activity in the chromosomal aberration assay. Compound 25 was identified as having the optimal balance of CXCR3 activity and pharmacokinetic properties across multiple pre-clinical species, which are reported herein.
Asunto(s)
Quinazolinas/síntesis química , Quinazolinonas/síntesis química , Receptores CXCR3/antagonistas & inhibidores , Animales , Bleomicina/toxicidad , Aberraciones Cromosómicas , Citocromo P-450 CYP3A , Inhibidores del Citocromo P-450 CYP3A , Perros , Relación Dosis-Respuesta a Droga , Diseño de Fármacos , Humanos , Inflamación , Concentración 50 Inhibidora , Leucocitos/efectos de los fármacos , Macaca fascicularis , Ratones , Modelos Químicos , Quinazolinas/farmacología , Quinazolinonas/farmacología , Factores de TiempoRESUMEN
Starting from a series of ureas that were determined to be mechanism-based inhibitors of FAAH, several spirocyclic ureas and lactams were designed and synthesized. These efforts identified a series of novel, noncovalent FAAH inhibitors with in vitro potency comparable to known covalent FAAH inhibitors. The mechanism of action for these compounds was determined through a combination of SAR and co-crystallography with rat FAAH.
Asunto(s)
Amidohidrolasas/antagonistas & inhibidores , Inhibidores Enzimáticos/química , Amidohidrolasas/metabolismo , Animales , Sitios de Unión , Simulación por Computador , Cristalografía por Rayos X , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/farmacocinética , Humanos , Lactamas/síntesis química , Lactamas/química , Lactamas/farmacocinética , Ratas , Compuestos de Espiro/química , Relación Estructura-Actividad , Urea/síntesis química , Urea/química , Urea/farmacocinéticaRESUMEN
A general way of improving the potency of CXCR3 antagonists with fused hetero-bicyclic cores was identified. Optimization efforts led to the discovery of a series of imidazo-pyrazine derivatives with improved pharmacokinetic properties in addition to increased potency. The efficacy of the lead compound 21 is evaluated in a mouse lung inflammation model.
Asunto(s)
Antiinflamatorios/química , Bencenoacetamidas/química , Óxidos S-Cíclicos/química , Imidazoles/química , Pirazinas/química , Receptores CXCR3/antagonistas & inhibidores , Animales , Antiinflamatorios/síntesis química , Antiinflamatorios/farmacocinética , Bencenoacetamidas/síntesis química , Bencenoacetamidas/farmacocinética , Óxidos S-Cíclicos/síntesis química , Óxidos S-Cíclicos/farmacocinética , Humanos , Imidazoles/síntesis química , Imidazoles/farmacocinética , Ratones , Conformación Molecular , Pirazinas/síntesis química , Pirazinas/farmacocinética , Ratas , Receptores CXCR3/metabolismoRESUMEN
A series of six-six and six-five fused heterocyclic CXCR3 antagonists has been synthesized and their activities evaluated in an [(125)I]-IP-10 displacement assay and an ITAC mediated in vitro cell migration assay. The pharmacokinetic properties of several top compounds have also been studied. This effort led to the discovery of compounds with increased potency and improved pharmacokinetic properties that could serve as useful tools to study the role of the CXCR3 receptor in vivo.
Asunto(s)
Compuestos Heterocíclicos/farmacología , Quinazolinonas/farmacología , Receptores CXCR3/antagonistas & inhibidores , Animales , Cromatografía Líquida de Alta Presión , Humanos , Quinazolinonas/farmacocinética , Ratas , EstereoisomerismoRESUMEN
The first nonpeptidic, noncovalent inhibitors of the cysteine protease cathepsin S (CatS) are described. Electronic database searching using the program DOCK generated a screening set of potential CatS inhibitors from which two lead structures were identified as promising starting points for a drug discovery effort. Lead optimization afforded potent (IC(50) < 50 nM) and selective inhibitors of CatS demonstrating cellular activity and reversibility of enzyme inhibition.
Asunto(s)
Catepsinas/antagonistas & inhibidores , Inhibidores de Cisteína Proteinasa/química , Inhibidores de Cisteína Proteinasa/farmacología , Linfocitos B/efectos de los fármacos , Linfocitos B/metabolismo , Catepsinas/química , Células Cultivadas , Evaluación Preclínica de Medicamentos/métodos , Antígenos de Histocompatibilidad Clase II/efectos de los fármacos , Antígenos de Histocompatibilidad Clase II/metabolismo , Humanos , Regiones Constantes de Inmunoglobulina/efectos de los fármacos , Regiones Constantes de Inmunoglobulina/metabolismo , Concentración 50 Inhibidora , Péptidos/química , Péptidos/farmacología , Conformación Proteica , Relación Estructura-ActividadRESUMEN
A series of competitive, reversible cathepsin S (CatS) inhibitors was investigated. An earlier disclosure detailed the discovery of the 4-(2-keto-1-benzimidazolinyl)-piperidin-1-yl moiety as an effective replacement for the 4-arylpiperazin-1-yl group found in our screening hit. Continued investigation into replacements for the 4-aryl piperazine resulted in the identification of potentially useful CatS inhibitors with enzymatic and cellular activity similar to that of JNJ 10329670 as disclosed in a previous publication.
Asunto(s)
Compuestos Bicíclicos con Puentes/química , Catepsinas/antagonistas & inhibidores , Inhibidores Enzimáticos/química , Piperidinas/química , Animales , Sitios de Unión , Compuestos Bicíclicos con Puentes/farmacología , Línea Celular Tumoral , Inhibidores Enzimáticos/farmacología , Ratones , Piperidinas/farmacología , Relación Estructura-ActividadRESUMEN
Aldol reactions of methyl ketone lithium enolates proceed via chairlike Zimmerman-Traxler transition states with 7:1 to 50:1 preference over alternative, boatlike transition structures, as determined by studies involving the configurationally stable deuterium-labeled enol silane 18 as the lithium enolate precursor.
Asunto(s)
Alcoholes/síntesis química , Aldehídos/química , Cetonas/síntesis química , Litio/química , Pentanonas/química , Deuterio , Cetonas/química , Conformación Molecular , Compuestos Organometálicos/química , EstereoisomerismoRESUMEN
A novel series of competitive, reversible cathepsin S (CatS) inhibitors was discovered and optimized. The 4-(2-keto-1-benzimidazolinyl)-piperidin-1-yl moiety was found to be an effective replacement for the 4-arylpiperazin-1-yl group found in our earlier series of CatS inhibitors. This replacement imparted improved PK properties as well as decreased off-target activity. Optimization of the ketobenzimidazole moiety led to the discovery of the lead compound JNJ 10329670, which represents a novel class of selective, noncovalent, reversible, and orally bioavailable inhibitors of cathepsin S.
Asunto(s)
Catepsinas/antagonistas & inhibidores , Animales , Bencimidazoles/química , Bencimidazoles/farmacocinética , Bencimidazoles/farmacología , Línea Celular , Compuestos Heterocíclicos con 2 Anillos/química , Compuestos Heterocíclicos con 2 Anillos/farmacología , Modelos Químicos , Estructura Molecular , Pirazoles/química , Pirazoles/farmacocinética , Pirazoles/farmacología , Piridinas/química , Piridinas/farmacocinética , Piridinas/farmacología , Relación Estructura-ActividadRESUMEN
We describe a simple 1H NMR analysis that permits the stereochemistry of beta-hydroxy ketones to be assigned by visual inspection of the ABX patterns for the alpha-methylene unit of the beta-hydroxy ketone in the 1H NMR spectra. This method has been verified by application to a wide range of beta-hydroxy ketones deriving from aldol reactions of chiral aldehydes with a variety of chiral and achiral methyl ketone enolates (see Tables 1 and 2). The stereochemistry of 54 of these compounds have been assigned by rigorous chemical methods.