RESUMEN
Complement C1s protease inhibitors have potential utility in the treatment of diseases associated with activation of the classical complement pathway such as humorally mediated graft rejection, ischemia-reperfusion injury (IRI), vascular leak syndrome, and acute respiratory distress syndrome (ARDS). The utility of biphenylsulfonyl-thiophene-carboxamidine small-molecule C1s inhibitors are limited by their poor in vivo pharmacokinetic properties. Pegylation of a potent analog has provided compounds with good potency and good in vivo pharmacokinetic properties.
Asunto(s)
Amidas/química , Complemento C1s/antagonistas & inhibidores , Diseño de Fármacos , Polietilenglicoles/química , Inhibidores de Proteasas/síntesis química , Tiofenos/química , Animales , Complemento C1s/metabolismo , Semivida , Inhibidores de Proteasas/química , Inhibidores de Proteasas/farmacocinética , RatasRESUMEN
Complement activation has been implicated in disease states such as hereditary angioedema, ischemia-reperfusion injury, acute respiratory distress syndrome, and acute transplant rejection. Even though the complement cascade provides several protein targets for potential therapeutic intervention only two complement inhibitors have been approved so far for clinical use including anti-C5 antibodies for the treatment of paroxysmal nocturnal hemoglobinuria and purified C1-esterase inhibitor replacement therapy for the control of hereditary angioedema flares. In the present study, optimization of potency and physicochemical properties of a series of thiophene amidine-based C1s inhibitors with potential utility as intravenous agents for the inhibition of the classical pathway of complement is described.
Asunto(s)
Complemento C1s/antagonistas & inhibidores , Compuestos Heterocíclicos/química , Compuestos Heterocíclicos/farmacología , Animales , Sitios de Unión , Semivida , Modelos Moleculares , Estructura Molecular , Ratas , Relación Estructura-ActividadRESUMEN
HDM2 binds to an alpha-helical transactivation domain of p53, inhibiting its tumor suppressive functions. A miniaturized thermal denaturation assay was used to screen chemical libraries, resulting in the discovery of a novel series of benzodiazepinedione antagonists of the HDM2-p53 interaction. The X-ray crystal structure of improved antagonists bound to HDM2 reveals their alpha-helix mimetic properties. These optimized molecules increase the transcription of p53 target genes and decrease proliferation of tumor cells expressing wild-type p53.
Asunto(s)
Benzodiazepinas/síntesis química , Proteínas Nucleares/antagonistas & inhibidores , Proteínas Proto-Oncogénicas/antagonistas & inhibidores , Proteína p53 Supresora de Tumor/agonistas , Benzodiazepinas/química , Benzodiazepinas/farmacología , Sitios de Unión , Línea Celular Tumoral , Técnicas Químicas Combinatorias , Cristalografía por Rayos X , Humanos , Modelos Moleculares , Imitación Molecular , Estructura Molecular , Proteínas Proto-Oncogénicas c-mdm2 , Estereoisomerismo , Relación Estructura-Actividad , Proteína p53 Supresora de Tumor/biosíntesisRESUMEN
We have identified RWJ-671818 (8) as a novel, low molecular weight, orally active inhibitor of human alpha-thrombin (K(i) = 1.3 nM) that is potentially useful for the acute and chronic treatment of venous and arterial thrombosis. In a rat deep venous thrombosis model used to assess antithrombotic efficacy, oral administration of 8 at 30 and 50 mg/kg reduced thrombus weight by 87 and 94%, respectively. In an anesthetized rat antithrombotic model, where electrical stimulation of the carotid artery created a thrombus, 8 prolonged occlusion time 2- and 3-fold at 0.1 and 1.0 mg/kg, i.v., respectively, and more than doubled activated clotting time and activated partial thromboplastin time at the higher dose. This compound had excellent oral bioavailability of 100% in dogs with an estimated half-life of approximately 3 h. On the basis of its noteworthy preclinical data, 8 was advanced into human clinical trials and successfully progressed through phase 1 studies.
Asunto(s)
Anticoagulantes/síntesis química , Fibrinolíticos/síntesis química , Guanidinas/síntesis química , Pirazinas/síntesis química , Trombina/antagonistas & inhibidores , Secuencias de Aminoácidos , Animales , Anticoagulantes/farmacocinética , Anticoagulantes/farmacología , Presión Sanguínea/efectos de los fármacos , Células CACO-2 , Cristalografía por Rayos X , Citocromo P-450 CYP3A , Inhibidores del Citocromo P-450 CYP3A , Perros , Método Doble Ciego , Electrocardiografía , Femenino , Fibrinolíticos/farmacocinética , Fibrinolíticos/farmacología , Guanidinas/farmacocinética , Guanidinas/farmacología , Cobayas , Frecuencia Cardíaca/efectos de los fármacos , Hemodinámica/efectos de los fármacos , Humanos , Técnicas In Vitro , Masculino , Microsomas Hepáticos/metabolismo , Modelos Moleculares , Pirazinas/farmacocinética , Pirazinas/farmacología , Ratas , Ratas Sprague-Dawley , Proteínas Recombinantes/antagonistas & inhibidores , Relación Estructura-Actividad , Trombina/química , Trombosis de la Vena/sangre , Trombosis de la Vena/tratamiento farmacológicoAsunto(s)
Discusiones Bioéticas , Bioética , Directivas Anticipadas , Altruismo , Beneficencia , Coerción , Confidencialidad , Toma de Decisiones , Deber de Advertencia , Ética Clínica , Ética Médica , Eutanasia Pasiva , Testimonio de Experto , Honorarios y Precios , Libertad , Ingeniería Genética , Seropositividad para VIH , Asignación de Recursos para la Atención de Salud , Proyecto Genoma Humano , Humanos , Consentimiento Informado , Jurisprudencia , Competencia Mental , Obligaciones Morales , Trasplante de Órganos , Atención al Paciente , Selección de Paciente , Autonomía Personal , Médicos , Diagnóstico Prenatal , Consentimiento Presumido , Negativa al Tratamiento , Asignación de Recursos , Órdenes de Resucitación , Justicia Social , Responsabilidad Social , Consentimiento por Terceros , Donantes de Tejidos , Obtención de Tejidos y Órganos , Negativa del Paciente al Tratamiento , Privación de TratamientoRESUMEN
The pharmacokinetics of TDP4815 was evaluated in rats, rabbits, dogs and monkeys. After intravenous administration, TDP4815 achieved C(O) of 3255 ng/ml in rats at 5 mg/kg, 9066 ng/ml in rabbits and 7858 ng/ml in monkeys at 6 mg/kg, and 4457 ng/ml in dogs at 3 mg/kg. The clearance (C(L)) was 3105, 1692, 835 and 640 ml/h/kg in rats, rabbits, monkeys and dogs, respectively. The volume of distribution (V(Z)) was more than 3861 ml/kg in all species, except 1915 ml/kg in monkeys. The oral bioavailability was rabbit >rat> monkey compared at 100 mg/kg, but it was much higher in dogs (>64%) after oral administrations. The calculated intrinsic clearance data suggested that the clearance of dog and human was restricted by binding to the plasma protein, and the clearance of rat and monkey was dependent on both the free fraction of plasma protein binding and the liver blood flow rate. The unbound hepatic intrinsic clearance of monkey was close to its C(L) suggesting that the hepatic clearance was an important excretion in monkeys. The poor oral bioavailability in the monkey may be related to the extensive glucuronidation. The V(Z).kg and C(L).kg in test species showed good correlation with the animal body weights (R(2)=0.87 and 0.96).
Asunto(s)
Anticoagulantes/farmacocinética , Guanidina/análogos & derivados , Administración Oral , Animales , Anticoagulantes/administración & dosificación , Disponibilidad Biológica , Peso Corporal , Perros , Glucurónidos/metabolismo , Guanidina/administración & dosificación , Guanidina/farmacocinética , Humanos , Técnicas In Vitro , Inyecciones Intravenosas , Hígado/irrigación sanguínea , Hígado/metabolismo , Macaca fascicularis , Masculino , Microsomas Hepáticos/metabolismo , Unión Proteica , Conejos , Ratas , Ratas Sprague-Dawley , Especificidad de la Especie , Distribución TisularRESUMEN
Inhibiting the classical pathway of complement activation by attenuating the proteolytic activity of the serine protease C1s is a potential strategy for the therapeutic intervention in disease states such as hereditary angioedema, ischemia-reperfusion injury, and acute transplant rejection. A series of arylsulfonylthiophene-2-carboxamidine inhibitors of C1s were synthesized and evaluated for C1s inhibitory activity. The most potent compound had a Ki of 10nM and >1000-fold selectivity over uPA, tPA, FX(a), thrombin, and plasmin.
Asunto(s)
Arilsulfonatos/síntesis química , Complemento C1s/antagonistas & inhibidores , Inhibidores de Serina Proteinasa/síntesis química , Amidinas/síntesis química , Amidinas/farmacología , Angioedema/tratamiento farmacológico , Arilsulfonatos/farmacología , Fibrinolisina/farmacología , Rechazo de Injerto/tratamiento farmacológico , Humanos , Isquemia Miocárdica/tratamiento farmacológico , Inhibidores de Serina Proteinasa/farmacología , Relación Estructura-Actividad , Acetato de Tetradecanoilforbol/farmacología , Tiofenos/síntesis química , Tiofenos/farmacología , Trombina/farmacología , Activador de Plasminógeno de Tipo Uroquinasa/farmacologíaRESUMEN
The protein product of an essential gene of unknown function from Streptococcus pneumoniae was expressed and purified for screening in the ThermoFluor affinity screening assay. This assay can detect ligand binding to proteins of unknown function. The recombinant protein was found to be in a dimeric, native-like folded state and to unfold cooperatively. ThermoFluor was used to screen the protein against a library of 3000 compounds that were specifically selected to provide information about possible biological functions. The results of this screen identified pyridoxal phosphate and pyridoxamine phosphate as equilibrium binding ligands (K(d) approximately 50 pM, K(d) approximately 2.5 microM, respectively), consistent with an enzymatic cofactor function. Several nucleotides and nucleotide sugars were also identified as ligands of this protein. Sequence comparison with two enzymes of known structure but relatively low overall sequence homology established that several key residues directly involved in pyridoxal phosphate binding were strictly conserved. Screening a collection of generic drugs and natural products identified the antifungal compound canescin A as an irreversible covalent modifier of the enzyme. Our investigation of this protein indicates that its probable biological role is that of a nucleoside diphospho-keto-sugar aminotransferase, although the preferred keto-sugar substrate remains unknown. These experiments demonstrate the utility of a generic affinity-based ligand binding technology in decrypting possible biological functions of a protein, an approach that is both independent of and complementary to existing genomic and proteomic technologies.
Asunto(s)
Proteínas Bacterianas/fisiología , Genes Esenciales/fisiología , Azúcares de Nucleósido Difosfato/metabolismo , Streptococcus pneumoniae/genética , Transaminasas/fisiología , Secuencia de Aminoácidos , Benzopiranos/metabolismo , Dimerización , Furanos/metabolismo , Ligandos , Datos de Secuencia Molecular , Fosfato de Piridoxal/metabolismo , Piridoxamina/metabolismo , Streptococcus pneumoniae/enzimologíaRESUMEN
Although thrombin has been extensively researched with many examples of potent and selective inhibitors, the key characteristics of oral bioavailability and long half-life have been elusive. We report here a novel series non-peptidic phenyl-based, highly potent, highly selective and orally bioavailable thrombin inhibitors using oxyguanidines as guanidine-mimetics.
Asunto(s)
Derivados del Benceno/química , Derivados del Benceno/farmacología , Guanidinas/química , Guanidinas/farmacología , Trombina/antagonistas & inhibidores , Administración Oral , Animales , Derivados del Benceno/farmacocinética , Disponibilidad Biológica , Células CACO-2 , Perros , Guanidinas/farmacocinética , Humanos , Ratones , Microsomas/efectos de los fármacos , Ratas , Serina Endopeptidasas/efectos de los fármacos , Inhibidores de Serina Proteinasa/farmacología , Relación Estructura-ActividadRESUMEN
Through structure-based drug design and parallel synthesis, we have discovered a novel series of nonpeptidic phenyl-based thrombin inhibitors using oxyguanidines as guanidine bioisosteres. These compounds have been found to be highly potent, highly selective, and orally bioavailable.
Asunto(s)
Anticoagulantes/síntesis química , Diseño de Fármacos , Guanidinas/síntesis química , Trombina/antagonistas & inhibidores , Administración Oral , Animales , Anticoagulantes/farmacocinética , Sitios de Unión , Línea Celular , Cristalografía por Rayos X , Perros , Guanidinas/farmacocinética , Humanos , Inhibidores de Serina Proteinasa/síntesis química , Inhibidores de Serina Proteinasa/farmacocinética , Relación Estructura-ActividadRESUMEN
Activation of the classical pathway of complement has been implicated in disease states such as hereditary angioedema, ischemia-reperfusion injury and acute transplant rejection. The trypsin-like serine protease C1s represents a pivotal upstream point of control in the classical pathway of complement activation and is therefore likely to be a useful target in the therapeutic intervention of these disease states. A series of thiopheneamidine-based inhibitors of C1s has been optimized to give a 70 nM inhibitor that inhibits the classical pathway of complement activation in vitro.
Asunto(s)
Proteínas Inactivadoras de Complemento/síntesis química , Vía Clásica del Complemento/efectos de los fármacos , Pirazoles/síntesis química , Serina Endopeptidasas/metabolismo , Inhibidores de Serina Proteinasa/síntesis química , Tiazoles/síntesis química , Tiofenos/síntesis química , Sitios de Unión/fisiología , Complemento C1/metabolismo , Proteínas Inactivadoras de Complemento/farmacología , Vía Clásica del Complemento/fisiología , Humanos , Isquemia Miocárdica/tratamiento farmacológico , Isquemia Miocárdica/enzimología , Pirazoles/farmacología , Inhibidores de Serina Proteinasa/farmacología , Relación Estructura-Actividad , Tiazoles/farmacología , Tiofenos/farmacologíaRESUMEN
A study of the S1 binding of lead 5-methylthiothiophene amidine 3, an inhibitor of urokinase-type plasminogen activator, was undertaken by the introduction of a variety of substituents at the thiophene 5-position. The 5-alkyl substituted and unsubstituted thiophenes were prepared using organolithium chemistry. Heteroatom substituents were introduced at the 5-position using a novel displacement reaction of 5-methylsulfonylthiophenes and the corresponding oxygen or sulfur anions. Small alkyl group substitution at the 5-position provided inhibitors equipotent with but possessing improved solubility.