RESUMEN
Anticoagulant agents have emerged as a promising class of therapeutic drugs for the treatment and prevention of arterial and venous thrombosis. We investigated a series of novel orally active factor Xa inhibitors designed using our previously reported conjugation strategy to boost oral anticoagulant effect. Structural optimization of anthranilamide derivative 3 as a lead compound with installation of phenolic hydroxyl group and extensive exploration of the P1 binding element led to the identification of 5-chloro-N-(5-chloro-2-pyridyl)-3-hydroxy-2-{[4-(4-methyl-1,4-diazepan-1-yl)benzoyl]amino}benzamide (33, AS1468240) as a potent factor Xa inhibitor with significant oral anticoagulant activity. We also reported a newly developed Free-Wilson-like fragment recommender system based on the integration of R-group decomposition with collaborative filtering for the structural optimization process.
Asunto(s)
Anticoagulantes/química , Inhibidores del Factor Xa/química , Factor Xa/química , Administración Oral , Animales , Anticoagulantes/síntesis química , Anticoagulantes/metabolismo , Benzamidas/síntesis química , Benzamidas/química , Benzamidas/metabolismo , Diseño de Fármacos , Factor Xa/metabolismo , Inhibidores del Factor Xa/síntesis química , Inhibidores del Factor Xa/metabolismo , Glucurónidos/química , Humanos , Masculino , Ratones , Ratones Endogámicos ICR , Unión Proteica , Tiempo de Protrombina , Relación Estructura-ActividadRESUMEN
The blood coagulation cascade represents an attractive target for antithrombotic drug development, and recent studies have attempted to identify oral anticoagulants with inhibitory activity for enzymes in this cascade, with particular attention focused on thrombin and factor Xa (fXa) as typical targets. We previously described the discovery of the orally active fXa inhibitor darexaban (1) and reported a unique profile that compound 1 rapidly transformed into glucuronide YM-222714 (2) after oral administration. Here, we propose a novel strategy towards the discovery of an orally active anticoagulant that is based on the bioconversion of a non-amidine inhibitor into the corresponding conjugate to boost ex vivo anticoagulant activity via an increase in hydrophilicity. Computational molecular modeling was utilized to select a template scaffold and design a substitution point to install a potential functional group for conjugation. This strategy led to the identification of the phenol-derived fXa inhibitor ASP8102 (14), which demonstrated highly potent anticoagulant activity after biotransformation into the corresponding glucuronide (16) via oral dosing.
Asunto(s)
Anticoagulantes/química , Inhibidores Enzimáticos/química , Factor Xa/química , Trombina/química , Administración Oral , Animales , Anticoagulantes/síntesis química , Anticoagulantes/farmacocinética , Azepinas/química , Azepinas/metabolismo , Benzamidas/química , Benzamidas/metabolismo , Sitios de Unión , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/farmacocinética , Factor Xa/metabolismo , Glucurónidos/química , Glucurónidos/metabolismo , Semivida , Macaca fascicularis , Simulación del Acoplamiento Molecular , Estructura Terciaria de Proteína , Tiempo de Protrombina , Trombina/metabolismoRESUMEN
Inhibitors of factor Xa (FXa), a crucial serine protease in the coagulation cascade, have attracted a great deal of attention as a target for developing antithrombotic agents. We previously reported findings from our optimization study of a high-throughput screening (HTS) derived lead compound 1a that resulted in the discovery of potent amidine-containing FXa inhibitors represented by compound 2. We also conducted an alternative optimization study of 1a without incorporating a strong basic amidine group, which generally has an adverse effect on the pharmacokinetic profile after oral administration. Replacement of 4-methoxybenzene with a 1,4-benzodiazepine structure and introduction of a hydroxy group at the central benzene led to the discovery of the potent and orally effective factor Xa inhibitor 14i (darexaban, YM150). Subsequent extensive study revealed a unique aspect to the pharmacokinetic profile of this compound, wherein the hydroxy moiety of 14i is rapidly transformed into its glucuronide conjugate 16 (YM-222714) as an active metabolite after oral administration and it plays a major role in expression of potent anticoagulant activity in plasma. The distinctive, potent activity of inhibitor 14i after oral dosing was explained by this unique pharmacokinetic profile and its favorable membrane permeability. Compound 14i is currently undergoing clinical development for prevention and treatment of thromboembolic diseases.
Asunto(s)
Azepinas/síntesis química , Benzamidas/síntesis química , Inhibidores del Factor Xa , Fibrinolíticos/síntesis química , Administración Oral , Animales , Anticoagulantes/síntesis química , Anticoagulantes/farmacocinética , Anticoagulantes/farmacología , Azepinas/química , Azepinas/farmacología , Benzamidas/química , Benzamidas/farmacología , Disponibilidad Biológica , Dominio Catalítico , Factor Xa/química , Fibrinolíticos/química , Fibrinolíticos/farmacología , Glucurónidos/metabolismo , Humanos , Técnicas In Vitro , Masculino , Ratones , Ratones Endogámicos ICR , Microsomas Hepáticos/metabolismo , Modelos Moleculares , Ratas , Ratas Endogámicas F344 , Relación Estructura-ActividadRESUMEN
Eltrombopag, an orally-active small molecule thrombopoietin (TPO) receptor agonist, was used for the first time in 2008 to treat patients with chronic idiopathic thrombocytopenic purpura. Here, we investigated the pharmacological effect of a new orally-active small molecule TPO receptor agonist which may be effective in treating these patients. 50% effective concentration values for cell proliferation with AS1670542 or eltrombopag were 1.9 and 13nM, respectively, while those for megakaryocyte colony formation from human cord blood CD34(+) cells with AS1670542 or eltrombopag were 260 and 950nM, respectively. On Day 14 after the start of administration, AS1670542 significantly increased the number of human platelets in non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice with transplanted human hematopoietic stem cells at 0.3 (P<0.05); in contrast, while administration of eltrombopag also increased the numbers of these platelets at 30mg/kg/day (P=0.058), no statistical significance was noted in the increase. Here, we identified AS1670542, a novel orally-active TPO receptor agonist which mimics the biological activity of TPO and may demonstrate greater in vitro and in vivo pharmacologically efficacy than eltrombopag.
Asunto(s)
Receptores de Trombopoyetina/agonistas , Tiazoles/administración & dosificación , Tiazoles/farmacología , Tiofenos/administración & dosificación , Tiofenos/farmacología , Administración Oral , Secuencia de Aminoácidos , Animales , Antígenos CD34/metabolismo , Benzoatos/farmacología , Línea Celular , Membrana Celular/metabolismo , Perros , Femenino , Histidina/metabolismo , Humanos , Hidrazinas/farmacología , Hígado/citología , Hígado/efectos de los fármacos , Hígado/metabolismo , Ratones , Datos de Secuencia Molecular , Recuento de Plaquetas , Estructura Terciaria de Proteína , Pirazoles/farmacología , Ratas , Receptores de Trombopoyetina/química , Receptores de Trombopoyetina/metabolismo , Especificidad de la Especie , Trasplante de Células MadreRESUMEN
Thrombopoietin (TPO) is the principal physiologic regulator of platelet production. We have searched for small molecule compounds that mimic the action of TPO by using human TPO receptor-expressed in Ba/F3 cells, resulting in the discovery of AKR-501 (YM477). AKR-501 specifically targeted the TPO receptor and stimulated megakaryocytopoiesis throughout the development and maturation of megakaryocytes just as rhTPO did. AKR-501, however, was shown to be effective only in humans and chimpanzees with high species specificity. Therefore, we examined the in vivo platelet-increasing effect of AKR-501 in human platelet producing non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice transplanted with human fetal liver CD34(+) cells. Daily oral administration of AKR-501 dose-dependently increased the number of human platelets in these mice, with significance achieved at doses of 1 mg/kg and above. The peak unbound plasma concentrations of AKR-501 after administration at 1 mg/kg in NOD/SCID mice were similar to those observed following administration of an active oral dose in human subjects. These results suggest that AKR-501 is an orally-active TPO receptor agonist that may be useful in the treatment of patients with thrombocytopenia.
Asunto(s)
Plaquetas/fisiología , Megacariocitos/fisiología , Receptores de Trombopoyetina/agonistas , Tiazoles/farmacología , Tiofenos/farmacología , Administración Oral , Animales , Antígenos CD34/fisiología , Plaquetas/citología , Plaquetas/efectos de los fármacos , Diferenciación Celular , Sangre Fetal/citología , Hematopoyesis/efectos de los fármacos , Hematopoyesis/fisiología , Humanos , Hígado/embriología , Megacariocitos/citología , Megacariocitos/efectos de los fármacos , Ratones , Ratones Endogámicos NOD , Ratones SCID , Pan troglodytes , Recuento de Plaquetas , Ploidias , Poliploidía , Receptores de Trombopoyetina/genética , Receptores de Trombopoyetina/fisiología , Proteínas Recombinantes/farmacología , Trasplante de Células Madre , Tiazoles/administración & dosificación , Tiofenos/administración & dosificación , Trombopoyetina/genética , Trombopoyetina/farmacologíaRESUMEN
OBJECTIVE: AKR-501 (YM477) is an orally active thrombopoietin (TPO) receptor agonist that mimics the biological effect of TPO in vitro and in vivo. Here, we report that AKR-501 in combination with TPO has additive effect on megakaryocytopoiesis. MATERIALS AND METHODS: Granulocyte colony-stimulating factor-mobilized human peripheral blood CD34+ cells were cultured with AKR-501, TPO, or a combination of the two in serum-free liquid culture system. The numbers of hematopoietic progenitor cells, megakaryocytic progenitor cells, and megakaryocytes were measured using flow cytometry. Further, the effect of AKR-501 on TPO binding to TPO receptor was examined. RESULTS: Both AKR-501 and TPO alone increased the number of megakaryocytes, and the maximum activities of AKR-501 and TPO were similar. Interestingly, in the presence of TPO concentrations producing maximal stimulation, the addition of AKR-501 increased the number of megakaryocytes to about 200% of that generated with TPO only. In the time course experiment, the combination of AKR-501 and TPO augmented the numbers of hematopoietic progenitor cells and colony-forming unit in culture in the early stages. Thus, the combination of AKR-501 and TPO enhanced not only the differentiation into megakaryocytes, but also the expansion of human hematopoietic progenitor cells. Further, AKR-501 did not inhibit TPO binding to the TPO receptor. This result indicated the possibility that AKR-501 and TPO may act simultaneously on the TPO receptor, and this could be responsible for their additive effect of on megakaryocytopoiesis. CONCLUSIONS: This study suggests that AKR-501 would be useful for the treatment of thrombocytopenia even at high plasma levels of endogenous TPO following chemotherapy.
Asunto(s)
Piperidinas/uso terapéutico , Receptores de Trombopoyetina/agonistas , Tiazoles/uso terapéutico , Tiofenos/uso terapéutico , Trombocitopenia/tratamiento farmacológico , Trombopoyesis/efectos de los fármacos , Trombopoyetina/uso terapéutico , Antígenos CD/sangre , Antígenos CD34/sangre , Plaquetas/efectos de los fármacos , Plaquetas/fisiología , Células Cultivadas , Ensayo de Unidades Formadoras de Colonias , Citometría de Flujo , Humanos , Trombopoyesis/fisiologíaRESUMEN
We describe here our investigation of a new series of orally active fXa inhibitors based on a prodrug strategy. Solid-phase parallel synthesis identified a unique series of fXa inhibitors with a substituted benzenesulfonyl group as a novel S4 binding element. This series resulted in compound 39, which exhibited potent inhibitory activity against fXa (IC50 = 13 nM) and excellent selectivity over thrombin (>7000-fold). The masking of its highly hydrophilic groups led to the creation of related prodrug 28, which demonstrated an anticoagulant effect after oral dosing.
Asunto(s)
Inhibidores del Factor Xa , Profármacos/síntesis química , Profármacos/farmacología , Inhibidores de Serina Proteinasa/síntesis química , Inhibidores de Serina Proteinasa/farmacología , Animales , Evaluación Preclínica de Medicamentos , Humanos , Espectroscopía de Resonancia Magnética , Masculino , Ratones , Ratones Endogámicos ICR , Modelos Moleculares , Profármacos/química , Inhibidores de Serina Proteinasa/química , Espectrometría de Masa Bombardeada por Átomos VelocesRESUMEN
We found the novel selective and orally available non-amidine TF/FVIIa complex inhibitor 21e, 4-({[(1S)-(aminocarbonyl)-3-methylbutyl]amino}carbonyl)-2'-({[4- (aminomethyl)phenyl]amino}carbonyl)-4'-(methylamino)biphenyl-2- carboxylic acid. The derivatives were synthesized by conversions of the isobutyl moiety and the introduction of alkylamino groups to 4'-position of the central phenyl ring of compounds 2a and 2b reported previously. Some compounds show increased in vitro anti-TF/FVIIa and PT prolongation activities. Among them, compound 21e reached and sustained micromolar plasma concentration levels of up to 2h after oral administration in mice. Moreover, compound 21e did not prolong the bleeding time even at the highest dose level in cynomolgus monkeys, while PT was prolonged 3.7-fold increases at this dose.
Asunto(s)
Compuestos de Bifenilo/síntesis química , Compuestos de Bifenilo/farmacología , Inhibidores de Factor de Coagulación Sanguínea/síntesis química , Factor VIIa/antagonistas & inhibidores , Lipoproteínas/síntesis química , Metilaminas/síntesis química , Metilaminas/farmacología , Tromboplastina/antagonistas & inhibidores , Administración Oral , Animales , Sitios de Unión , Compuestos de Bifenilo/química , Inhibidores de Factor de Coagulación Sanguínea/química , Inhibidores de Factor de Coagulación Sanguínea/farmacología , Diseño de Fármacos , Evaluación Preclínica de Medicamentos , Humanos , Enlace de Hidrógeno , Ligandos , Lipoproteínas/química , Lipoproteínas/farmacología , Macaca fascicularis , Masculino , Metilaminas/química , Ratones , Ratones Endogámicos ICR , Modelos Moleculares , Estructura Molecular , Estructura Secundaria de Proteína , Sensibilidad y Especificidad , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
Inhibition of tissue factor/factor VIIa complex (TF/FVIIa) is an attractive strategy for antithrombotic therapies. We began with an investigation of a non-amidine TF/FVIIa inhibitor based on a modification of amidine compound 1. Optimization of the substituents on the P1 phenyl portion of the compound 1 led to a neutral or less basic alternative for the 4-amidinophenyl moiety. By further optimization of the substituents on the central phenyl ring, a highly potent and selective TF/FVIIa inhibitor 17d was discovered.
Asunto(s)
Factor VIIa/antagonistas & inhibidores , Fibrinolíticos/química , Tromboplastina/antagonistas & inhibidores , Amidinas/química , Derivados del Benceno , Humanos , Ligandos , Unión Proteica/efectos de los fármacos , Relación Estructura-ActividadRESUMEN
A practical and cost-effective procedure has been developed for the synthesis of 7-methyl-2-naphthalenecarbonitrile, the precursor of the anticoagulant agents YM-60828 or YM-96765. This new route generates the key intermediate in only two steps from readily available 3-cyanopropionaldehyde dimethyl acetal and m-tolualdehyde, without requiring chromatographic purification. The synthesis involves condensation of the cyano derivative with the aldehyde and subsequent cyclodehydration.
Asunto(s)
Anticoagulantes/síntesis química , Naftalenos/síntesis química , Nitrilos/síntesis química , Benzaldehídos , Cromatografía en Capa Delgada , Compuestos Heterocíclicos/síntesis química , Espectroscopía de Resonancia Magnética , Piperidinas/síntesis química , Espectrofotometría UltravioletaRESUMEN
Factor Xa (fXa) is a serine protease that plays a pivotal role in the coagulation cascade. High-throughput screening of the Yamanouchi compound library yielded lead compound 1 with the ability to inhibit fXa at micromolar concentrations. To improve its fXa inhibitory activity and its oral anticoagulant activity, the linker between benzamidine and the central benzene ring was modified and a carboxyl group was introduced at the central benzene ring. The resulting compounds 40b (YM-203552), 41a (YM-202054), and 41c (YM-203558) exhibited potent fXa inhibitory activity and oral anticoagulant activity. In particular, YM-203558 exhibited the most potent oral anticoagulant activity, prolonging PT more than 3-fold at 0.5 and 2.0 h. Additionally, these compounds showed a high degree of selectivity for other serine proteases.
Asunto(s)
Derivados del Benceno/síntesis química , Derivados del Benceno/farmacología , Inhibidores del Factor Xa , Inhibidores de Serina Proteinasa/síntesis química , Inhibidores de Serina Proteinasa/farmacología , Animales , Derivados del Benceno/química , Espectroscopía de Resonancia Magnética , Masculino , Ratones , Modelos Moleculares , Inhibidores de Serina Proteinasa/química , Espectrometría de Masa Bombardeada por Átomos VelocesRESUMEN
Factor Xa (fXa) is a serine protease, which plays a pivotal role in the coagulation cascade. To improve the oral anticoagulant activity of fXa inhibitors containing a 1,4-diazepane moiety as the P4 part, a prodrug strategy was examined. Among the compounds evaluated in this study, amidoxime prodrugs bearing an ester moiety, such as compounds 21 and 30, showed effective oral anticoagulant activity in mice.
Asunto(s)
Amidinas/síntesis química , Amidinas/farmacología , Anticoagulantes/síntesis química , Anticoagulantes/farmacología , Antitrombina III/química , Profármacos/síntesis química , Profármacos/farmacología , Administración Oral , Amidinas/química , Animales , Anticoagulantes/química , Antitrombina III/síntesis química , Antitrombina III/farmacología , Compuestos Aza/síntesis química , Compuestos Aza/química , Compuestos Aza/farmacología , Pruebas de Coagulación Sanguínea , Femenino , Humanos , Ratones , EmbarazoRESUMEN
Factor Xa (fXa) is a serine protease involved in the coagulation cascade, which has received great interest as a potential target for the development of new antithrombotic drugs. Herein we report a novel series of fXa inhibitors in which the 1,4-diazepane moiety was designed to interact with the S4 aryl-binding domain of the fXa active site. Compound 13 (YM-96765) showed potent fXa inhibitory activity (IC(50) = 6.8 nM) and effective antithrombotic activity without prolonging bleeding time.
Asunto(s)
Compuestos Aza/química , Inhibidores de Serina Proteinasa/química , Animales , Compuestos Aza/síntesis química , Compuestos Aza/farmacología , Perros , Masculino , Ratones , Ratas , Ratas Sprague-Dawley , Inhibidores de Serina Proteinasa/síntesis química , Inhibidores de Serina Proteinasa/farmacologíaRESUMEN
Compound YM-60828 was previously characterized in our laboratory as a potent, selective and orally-bioavailable Factor Xa (FXa) inhibitor. The L-shape conformation of this compound in the active site of FXa was recognized as an important factor in displaying its FXa inhibitory activity. This led to the exploration of conformationally restricted cyclic scaffolds bearing a similar active conformation. The current study investigated a novel series of benzothiadiazine-4-one based compounds as FXa inhibitors. Structure-activity relationship (SAR) investigations revealed some potent FXa inhibitors that were selected for further in vitro and ex vivo anticoagulant studies. Among them, compound 6j (YM-169920) was proved to be most effective anticoagulant in this series. The synthesis and SAR in addition to docking studies of this class of inhibitors are described.
Asunto(s)
Benzotiadiazinas/química , Benzotiadiazinas/farmacología , Inhibidores del Factor Xa , Naftalenos/química , Naftalenos/farmacología , Piperidinas/química , Piperidinas/farmacología , Administración Oral , Animales , Anticoagulantes/síntesis química , Anticoagulantes/química , Anticoagulantes/farmacología , Benzotiadiazinas/síntesis química , Disponibilidad Biológica , Femenino , Humanos , Concentración 50 Inhibidora , Masculino , Ratones , Ratones Endogámicos ICR , Modelos Moleculares , Conformación Molecular , Naftalenos/síntesis química , Piperidinas/síntesis química , Tiempo de Protrombina , Saimiri , Relación Estructura-Actividad , Trombina/antagonistas & inhibidores , Inhibidores de Tripsina/síntesis química , Inhibidores de Tripsina/química , Inhibidores de Tripsina/farmacologíaRESUMEN
Factor Xa (FXa) is a serine protease which plays a pivotal role in the coagulation cascade. The inhibition of FXa has received great interest as a potential target for the development of new antithrombotic drug. Herein we describe a series of novel 7-amidino-2-naphthoanilide and 7-amidino-2-naphthalensulfonanilide derivatives which are potent FXa inhibitors. These scaffolds are rigid and are allowed to adopt an L-shape conformation which was estimated as the active conformation based on a docking study of YM-60828 with FXa. Optimization of the side chain at the central aniline nitrogen of 7-amidino-2-naphthoanilide has led to several potent and orally active FXa inhibitors. 5h (YM-169964), the best compound of these series, showed potent FXa inhibitory activity (IC(50)=3.9nM) and effectively prolonged prothrombin time by 9.6-fold ex vivo at an oral dose of 3mg/kg in squirrel monkeys.
Asunto(s)
Anilidas/síntesis química , Anticoagulantes/síntesis química , Inhibidores del Factor Xa , Administración Oral , Anilidas/farmacocinética , Anilidas/farmacología , Animales , Anticoagulantes/farmacocinética , Anticoagulantes/farmacología , Disponibilidad Biológica , Diseño de Fármacos , Femenino , Masculino , Ratones , Naftalenos/síntesis química , Naftalenos/farmacocinética , Naftalenos/farmacología , Piperidinas/síntesis química , Piperidinas/farmacocinética , Piperidinas/farmacología , Tiempo de Protrombina , Saimiri , Inhibidores de Serina Proteinasa/síntesis química , Inhibidores de Serina Proteinasa/farmacocinética , Inhibidores de Serina Proteinasa/farmacología , Relación Estructura-ActividadRESUMEN
Since Factor Xa (FXa) is well known to play a central role in thrombosis and hemostasis, inhibition of FXa is an attractive target for antithrombotic strategies. As a part of our investigation of a non-peptide, orally available FXa inhibitor, we found that a series of N-[(7-amidino-2-naphthyl)methyl]aniline derivatives possessed potent and selective inhibitory activities. Structure--activity relationship (SAR) of the substituent (R(1)) on the central aniline moiety suggested that increasing lipophilicity caused a detrimental effect on anticoagulant activity (prothrombin time assay) in plasma. Several compounds bearing a hydrophilic substituent in R(1) showed not only potent FXa inhibitory activities but also high anticoagulant activities. The best compound in this series was sulfamoylacetic acid derivative (YM-60828) which was a potent, selective and orally bioavailable FXa inhibitor and was chosen for clinical development.