RESUMEN
Based on insight from the X-ray crystal structure of human chymase in complex with compound 1, a lactam carbonyl of the diazepane core was exchanged with O-substituted oxyimino group, leading to amidoxime derivatives. This modification resulted in highly potent chymase inhibitors, such as O-phenylamidoxime 5f. X-ray crystal structure analysis indicated that compound 5f induced movement of the Leu99 and Tyr94 side chains at the S2 site, and the increase in inhibitory activity of O-phenyl amidoxime derivatives suggested that the O-phenyl moiety interacted with the Tyr94 residue. Surface plasmon resonance experiments showed that compound 5f had slower association and dissociation kinetics and the calculated residence time of compound 5f to human chymase was extended compared to that of amide compound 1.
Asunto(s)
Quimasas/antagonistas & inhibidores , Diseño de Fármacos , Oximas/farmacología , Inhibidores de Serina Proteinasa/farmacología , Sitios de Unión/efectos de los fármacos , Quimasas/metabolismo , Relación Dosis-Respuesta a Droga , Humanos , Estructura Molecular , Oximas/síntesis química , Oximas/química , Inhibidores de Serina Proteinasa/síntesis química , Inhibidores de Serina Proteinasa/química , Relación Estructura-ActividadRESUMEN
Novel thienopyrimidine compounds 2 and 3 were discovered from high-throughput screening as Natriuretic Peptide Receptor A (NPR-A) agonists. Scaffold hopping of a thienopyrimidine ring to a quinazoline ring, introduction of the basic functional group and optimization of the substituent on the 6-position of the benzene ring of quinazoline led to improved agonistic activity. We discovered compound 48, which showed potent agonistic activity for NPR-A with an EC50 value of 0.073µM, indicating 350-fold potency compared to the hit compound 3.
Asunto(s)
Pirimidinas/metabolismo , Receptores del Factor Natriurético Atrial/agonistas , Animales , Células CHO , Cricetinae , Cricetulus , Evaluación Preclínica de Medicamentos , Humanos , Pirimidinas/síntesis química , Pirimidinas/química , Quinazolinas/síntesis química , Quinazolinas/química , Quinazolinas/metabolismo , Receptores del Factor Natriurético Atrial/metabolismo , Relación Estructura-ActividadRESUMEN
Novel agonists of the Natriuretic Peptide Receptor A (NPR-A) were obtained through random screening and subsequent structural modification of triazine derivatives. The key structural feature to improve in vitro activity was the dimerization of triazine monomer derivatives. The non peptide derivative 7c and 13a showed highly potent NPR-A agonistic activity in vitro and diuretic activity in vivo. These results implied that non-peptidic small molecules open the possibility of new therapy for congestive heart failure.
Asunto(s)
Descubrimiento de Drogas , Receptores del Factor Natriurético Atrial/agonistas , Triazinas/farmacología , Animales , Cristalografía por Rayos X , GMP Cíclico/metabolismo , Dimerización , Diuréticos/farmacología , Humanos , Riñón/efectos de los fármacos , Riñón/metabolismo , Masculino , Espectrometría de Masas/métodos , Estructura Molecular , Espectroscopía de Protones por Resonancia Magnética , Ratas , Ratas Sprague-Dawley , Relación Estructura-Actividad , Triazinas/químicaRESUMEN
Natriuretic peptide receptor A (NPR-A) agonists were evaluated in vivo by optimizing the structure of quinazoline derivatives to improve agonistic activity for rat NPR-A. A 1,4-Cis-aminocyclohexylurea moiety at 4-position and hydroxy group of d-alaninol at 2-position on the quinazoline ring were found to be important factors in improving rat NPR-A activity. We identified potent quinazoline and pyrido[2,3-d]pyrimidine derivatives against rat NPR-A, with double-digit nanomolar EC50 values. The in vivo results showed that compound 56b administered at 1.0â¯mg/kg/min significantly increased plasma cGMP concentration and urine volume in rats. We discovered novel potent NPR-A agonists that showed agonistic effects similar to those of atrial natriuretic peptide.
Asunto(s)
Descubrimiento de Drogas , Quinazolinas/farmacología , Receptores del Factor Natriurético Atrial/agonistas , Animales , Relación Dosis-Respuesta a Droga , Humanos , Masculino , Estructura Molecular , Quinazolinas/síntesis química , Quinazolinas/química , Ratas , Ratas Sprague-Dawley , Relación Estructura-ActividadRESUMEN
A novel series of 6-benzyl substituted 4-aminocarbonyl-1,4-diazepane-2,5-diones were explored as human chymase inhibitors using structure-based drug design according to the X-ray cocrystal structure of chymase and compound 1. The optimization focused on the prime site led to the attainment of compounds that showed potent inhibitory activity, and among them, 18R shows a novel interaction mode.
Asunto(s)
Azepinas/síntesis química , Quimasas/antagonistas & inhibidores , Diseño de Fármacos , Inhibidores Enzimáticos/química , Etilmorfina/síntesis química , Azepinas/química , Azepinas/farmacología , Dominio Catalítico , Cristalografía por Rayos X , Activación Enzimática/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Etilmorfina/química , Etilmorfina/farmacología , Humanos , Concentración 50 Inhibidora , Estructura Molecular , Relación Estructura-ActividadRESUMEN
Our aim was to examine the effects of ASB17061, an orally active novel chymase inhibitor, on angiotensin II-induced abdominal aortic aneurysm (AAA) in apolipoprotein E-deficient mice. Oral administration of ASB17061 (10 mg/kg) significantly suppressed angiotensin II-induced AAA formation in these mice. The pro-matrix metalloproteinase-9 (pro-MMP-9) level in AAA lesions was significantly suppressed by ASB17061 treatment, indicating that ASB17061 inhibited the accumulation of pro-MMP-9-producing cells in AAA lesions. Mouse mast cell protease 4 (mMCP-4, human chymase ortholog) was injected into BALB/c mice intraperitoneally to examine the ability of mMCP-4 to induce the accumulation of pro-MMP-9-producing cells. An intraperitoneal injection of mMCP-4 induced the accumulation of pro-MMP-9-producing cells including CD11b + Gr-1 + cells. Taken together, these data indicate that ASB17061 is a promising novel oral therapeutic agent for human AAA.
Asunto(s)
Angiotensina II/farmacología , Aneurisma de la Aorta Abdominal/inducido químicamente , Aneurisma de la Aorta Abdominal/prevención & control , Apolipoproteínas E/deficiencia , Ácido Benzoico/farmacología , Quimasas/antagonistas & inhibidores , Inhibidores de Serina Proteinasa/farmacología , Animales , Aneurisma de la Aorta Abdominal/metabolismo , Colitis/prevención & control , Precursores Enzimáticos/metabolismo , Masculino , Metaloproteinasa 9 de la Matriz/metabolismo , Ratones , Ratones Endogámicos BALB CRESUMEN
The R5 subfamily of receptor-type protein tyrosine phosphatases (RPTPs) comprises PTPRZ and PTPRG. A recent study on primary human glioblastomas suggested a close association between PTPRZ1 (human PTPRZ) expression and cancer stemness. However, the functional roles of PTPRZ activity in glioma stem cells have remained unclear. In the present study, we found that sphere-forming cells from the rat C6 and human U251 glioblastoma cell lines showed high expression levels of PTPRZ-B, the short receptor isoform of PTPRZ. Stable PTPRZ knockdown altered the expression levels of stem cell transcription factors such as SOX2, OLIG2, and POU3F2 and decreased the sphere-forming abilities of these cells. Suppressive effects on the cancer stem-like properties of the cells were also observed following the knockdown of PTPRG. Here, we identified NAZ2329, a cell-permeable small molecule that allosterically inhibits both PTPRZ and PTPRG. NAZ2329 reduced the expression of SOX2 in C6 and U251 cells and abrogated the sphere-forming abilities of these cells. Tumor growth in the C6 xenograft mouse model was significantly slower with the co-treatment of NAZ2329 with temozolomide, an alkylating agent, than with the individual treatments. These results indicate that pharmacological inhibition of R5 RPTPs is a promising strategy for the treatment of malignant gliomas.
Asunto(s)
Carcinogénesis/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Glioblastoma/prevención & control , Células Madre Neoplásicas/efectos de los fármacos , Proteínas Tirosina Fosfatasas Clase 5 Similares a Receptores/antagonistas & inhibidores , Bibliotecas de Moléculas Pequeñas/farmacología , Temozolomida/farmacología , Animales , Antineoplásicos Alquilantes/farmacología , Carcinogénesis/metabolismo , Carcinogénesis/patología , Femenino , Glioblastoma/enzimología , Glioblastoma/patología , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Células Madre Neoplásicas/enzimología , Células Madre Neoplásicas/patología , Ratas , Proteínas Tirosina Fosfatasas Clase 5 Similares a Receptores/metabolismo , Células Tumorales Cultivadas , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Human chymase is known to function as a chemoattractant for human leukocytes. To investigate the mechanism of the chymase-induced cell migration, change in intracellular calcium concentration ([Ca(2+)]i) was examined in human polymorphonuclear (PMN) cells using Fluo-3 as a fluorescent Ca(2+) indicator. Treatment of PMN cells with human chymase caused [Ca(2+)]i elevation in a concentration-dependent manner. Depletion of extracellular Ca(2+) from the medium partially attenuated the chymase-induced [Ca(2+)]i increase, showing that both Ca(2+) influx and Ca(2+) release from internal stores might be involved in the [Ca(2+)]i response. Pretreatment of the cells with pertussis toxin completely blocked the chymase-induced [Ca(2+)]i signal, suggesting an involvement of G protein in the chymase-mediated [Ca(2+)]i elevation. The data in the present study raise the possibility that the chymase-induced cell migration is mediated by the [Ca(2+)]i elevation, which might be caused by stimulation of a G-protein-coupled receptor such as protease-activated receptors (PARs).
Asunto(s)
Señalización del Calcio/fisiología , Calcio/metabolismo , Neutrófilos/metabolismo , Serina Endopeptidasas/metabolismo , Señalización del Calcio/efectos de los fármacos , Quimasas , Inhibidores Enzimáticos/farmacología , Humanos , Neutrófilos/efectos de los fármacos , Toxina del Pertussis/farmacología , Serina Endopeptidasas/efectos de los fármacosRESUMEN
The possible role of mast cell chymase in organ fibrosis was examined using a bleomycin-induced pulmonary fibrosis model in mice. Intratracheal injection of bleomycin to mice significantly increased not only hydroxyproline content but also chymase activity in the lung. Administration of a chymase inhibitor SUN C8077 (7-chloro-3-(3-amynophenyl) quinazoline-2, 4-dione methanesulfonate) dose-dependently reversed the bleomycin-induced increase in hydroxyproline content as well as chymase activity in the lung. Human chymase digested latent transforming growth factor-beta1 (TGF-beta1) to form mature TGF-beta1 in vitro, which was inhibited by SUN C8077. Human chymase, on the other hand, failed to stimulate DNA synthesis of human lung fibroblasts CCD-8Lu and LL97A. Taken together, it is suggested that mast cell chymase might participate in the pathogenesis of pulmonary fibrosis, and that the chymase-induced fibrosis might be mediated at least in part by TGF-beta1. Chymase inhibitor may be promising for treatment of pulmonary fibrosis in humans.
Asunto(s)
Antibióticos Antineoplásicos , Bleomicina , Mastocitos/enzimología , Fibrosis Pulmonar/enzimología , Fibrosis Pulmonar/patología , Serina Endopeptidasas/fisiología , Animales , Biotransformación , Quimasas , Relación Dosis-Respuesta a Droga , Fibroblastos/efectos de los fármacos , Humanos , Hidroxiprolina/metabolismo , Pulmón/enzimología , Pulmón/patología , Masculino , Ratones , Ratones Endogámicos ICR , Mitógenos/farmacología , Fibrosis Pulmonar/inducido químicamente , Proteínas Recombinantes , Serina Endopeptidasas/farmacología , Factor de Crecimiento Transformador beta/fisiologíaRESUMEN
A chymase inhibitor SUN13834 has been shown to improve skin condition in animal models for atopic dermatitis. In the present study, effective dosages of SUN13834 for atopic dermatitis patients were predicted by pharmacokinetic/pharmacodynamic (PK/PD) analyses of SUN13834 in NC/Nga mice, which spontaneously develop atopic dermatitis-like skin lesions. For the PK/PD analyses, we utilized the minimum effective plasma concentration of unbound SUN13834 in late-phase reaction of trinitrochlorobenzene (TNCB)-induced biphasic dermatitis in mice, based on the assumption that the minimum effective plasma concentrations are the same among the two animal models. In late-phase reaction of biphasic dermatitis, SUN13834 was most effective when its plasma concentration was highest at the elicitation, and the minimum effective plasma concentration of unbound SUN13834 at the elicitation was calculated to be 0.13-0.2 ng/mL. Oral administration of SUN13834 improved dermatitis in NC/Nga mice at 15 mg/kg (twice a day; bid) and 30 mg/kg (once a day; qd), but not at 60 mg/kg (every other day; eod). At the three dosages, the duration times over the plasma level of 0.13-0.2 ng/mL were 16.1-20.3, 10.7-12.2 and 7.8-8.8h, respectively, suggesting an importance of maintenance of the minimum effective plasma concentration for at least about 10-12h. The clinical effective dosage predicted in this paper is also discussed in relation to a recently conducted Phase 2a study.
Asunto(s)
Azepinas/administración & dosificación , Quimasas/metabolismo , Dermatitis Atópica/tratamiento farmacológico , Inhibidores Enzimáticos/administración & dosificación , Piel/efectos de los fármacos , Administración Oral , Animales , Azepinas/farmacocinética , Ensayos Clínicos Fase II como Asunto , Dermatitis Atópica/inducido químicamente , Dermatitis Atópica/enzimología , Susceptibilidad a Enfermedades , Cálculo de Dosificación de Drogas , Inhibidores Enzimáticos/farmacocinética , Humanos , Ratones , Ratones Endogámicos , Cloruro de Picrilo/administración & dosificación , Piel/patologíaRESUMEN
Chymase is a chymotrypsin-like serine protease exclusively stored in secretory granules of mast cells and has been thought to participate in allergic diseases. It has already been shown that chymase inhibitor SUN13834 improves dermatitis in NC/Nga mice that spontaneously develop dermatitis resembling atopic dermatitis. In the present study, effect of chymase inhibitor SUN13834 on itch, the major feature of atopic dermatitis, was examined using a mouse dermatitis model induced by repeated topical application of 2,4-dinitrofluorobenzene (DNFB). Oral administration of SUN13834 once a day for 5 weeks inhibited not only skin swelling but accumulation of inflammatory cells including mast cells and eosinophils in the skin of the mice. In addition, SUN13834 also decreased significantly at 10 and 50 mg/kg the amount of scratching behavior induced by the DNFB challenge. This result indicates for the first time that mast cell chymase may be involved in itch induction. In conclusion, SUN13834 is thought to be useful as therapeutic agent for atopic dermatitis.
Asunto(s)
Azepinas/farmacología , Quimasas/antagonistas & inhibidores , Dermatitis Atópica/tratamiento farmacológico , Inhibidores Enzimáticos/farmacología , Administración Oral , Animales , Azepinas/administración & dosificación , Dermatitis Atópica/fisiopatología , Dinitrofluorobenceno , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Inhibidores Enzimáticos/administración & dosificación , Eosinófilos/efectos de los fármacos , Eosinófilos/metabolismo , Femenino , Inflamación/tratamiento farmacológico , Inflamación/etiología , Mastocitos/efectos de los fármacos , Mastocitos/metabolismo , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C3H , Prurito/tratamiento farmacológico , Prurito/etiología , Piel/efectos de los fármacos , Piel/patologíaRESUMEN
A novel series of 6-substituted 4-sulfonyl-1,4-diazepane-2,5-diones were designed, synthesized and evaluated as human chymase inhibitors. Structure-activity relationship studies led to the identification of a potent inhibitor, (6S)-6-(5-chloro-2-methoxybenzyl)-4-[(4-chlorophenyl)sulfonyl]-1,4-diazepane-2,5-dione, with an IC(50) of 0.027 microM.
Asunto(s)
Azepinas/farmacología , Quimasas/antagonistas & inhibidores , Inhibidores de Serina Proteinasa/farmacología , Azepinas/síntesis química , Humanos , Concentración 50 Inhibidora , Modelos Químicos , Inhibidores de Serina Proteinasa/síntesis química , Relación Estructura-ActividadRESUMEN
A novel series of 6-benzyl substituted 4-aminocarbonyl-1,4-diazepane-2,5-diones was designed, synthesized, and evaluated as human chymase inhibitors. From this series, we identified several compounds which were effective, via oral administration, in a mouse model of chronic dermatitis.
Asunto(s)
Azepinas/farmacología , Compuestos de Bencilo/farmacología , Quimasas/antagonistas & inhibidores , Dermatitis/tratamiento farmacológico , Inhibidores de Serina Proteinasa/farmacología , Administración Oral , Animales , Azepinas/síntesis química , Compuestos de Bencilo/síntesis química , Sitios de Unión , Dermatitis/patología , Modelos Animales de Enfermedad , Humanos , Concentración 50 Inhibidora , Ratones , Modelos Químicos , Inhibidores de Serina Proteinasa/síntesis química , Inhibidores de Serina Proteinasa/uso terapéutico , Relación Estructura-ActividadRESUMEN
The ability to convert angiotensin (Ang) I to Ang II was compared between human alpha-chymase and two mouse beta-chymases, mouse mast cell protease (mMCP)-1 and mMCP-4. Human chymase hydrolyzed Ang I to produce Ang II without further degradation. mMCP-1 similarly generated Ang II from Ang I in a time-dependent manner and the formation of the fragment other than Ang II was marginal. In contrast, mMCP-4 hydrolyzed Ang I at two sites, Tyr(4)-Ile(5) and Phe(8)-His(9), with Ang II formation being tentative. Consistently, mMCP-4 but not human chymase hydrolyzed Ang II and mMCP-1 showed little hydrolytic activity against Ang II. These data suggest that not only human chymase but also mMCP-1 might possess a physiological role in Ang II formation. Our findings also imply that the Ang-converting activity of chymase may not be related to the categorization of chymase into alpha- or beta-type based on their primary structure.