RESUMEN

Among cancer cells, indoleamine 2, 3-dioxygenase1 (IDO1) activity has been implicated in improving the proliferation and growth of cancer cells and suppressing immune cell activity. IDO1 is also responsible for the catabolism of tryptophan to kynurenine. Depletion of tryptophan and an increase in kynurenine exert important immunosuppressive functions by activating regulatory T cells and suppressing CD8+ T and natural killer (NK) cells. In this study, we compared the anti-tumor effects of YH29407, the best-in-class IDO1 inhibitor with improved pharmacodynamics and pharmacokinetics, with first and second-generation IDO1 inhibitors (epacadostat and BMS-986205, respectively). YH29407 treatment alone and anti-PD-1 (aPD-1) combination treatment induced significant tumor suppression compared with competing drugs. In particular, combination treatment showed the best anti-tumor effects, with most tumors reduced and complete responses. Our observations suggest that improved anti-tumor effects were caused by an increase in T cell infiltration and activity after YH29407 treatment. Notably, an immune depletion assay confirmed that YH29407 is closely related to CD8+ T cells. RNA-seq results showed that treatment with YH29407 increased the expression of genes involved in T cell function and antigen presentation in tumors expressing ZAP70, LCK, NFATC2, B2M, and MYD88 genes. Our results suggest that an IDO1 inhibitor, YH29407, has enhanced PK/PD compared to previous IDO1 inhibitors by causing a change in the population of CD8+ T cells including infiltrating T cells into the tumor. Ultimately, YH29407 overcame the limitations of the competing drugs and displayed potential as an immunotherapy strategy in combination with aPD-1.

RESUMEN

INTRODUCTION: Lazertinib is a potent, irreversible, brain-penetrant, mutant-selective, and wild-type-sparing third-generation EGFR tyrosine kinase inhibitor (TKI), creating a wide therapeutic index. Cardiovascular adverse events (AEs), including QT prolongation, decreased left ventricular ejection fraction (LVEF), and heart failure, have emerged as potential AEs with certain EGFR TKI therapies. METHODS: Cardiac safety of lazertinib was evaluated in TKI-tolerant adults with EGFR mutation-positive locally advanced or metastatic NSCLC receiving lazertinib (20-320 mg/d). QT intervals corrected with Fridericia's formula (QTcF) prolongation, time-matched concentration-QTcF relationship, change of LVEF, and cardiac failure-associated AEs were evaluated. The clinical findings were supplemented by the following three preclinical studies: an in vitro hERG inhibition assay, an ex vivo isolated perfused rabbit heart study, and an in vivo telemetry-instrumented beagle dog study. RESULTS: Preclinical evaluation revealed little to no physiological effect on the basis of electrocardiogram, electrophysiological, proarrhythmic, and hemodynamic parameters. Clinical evaluation of 181 patients revealed no clinically relevant QTcF prolongation by centralized electrocardiogram in any patient and at any dose level. The predicted magnitude of QTcF value increase at maximum steady-state plasma concentration for the therapeutic dose of lazertinib (240 mg/d) was 2.2 msec (upper bound of the two-sided 90% confidence interval: 3.6 msec). No patient had clinically relevant LVEF decrease (i.e., minimum postbaseline LVEF value of <50% and a maximum decrease in LVEF value from baseline of ≥10 percentage points). Cardiac failure-associated AE occurred in one patient (grade 2 decreased LVEF) and resolved without any dose modifications. CONCLUSIONS: Our first-in-human study, together with preclinical data, indicates that lazertinib is not associated with increased cardiac risk.

RESUMEN

PURPOSE: Given that osimertinib is the only approved third-generation EGFR-TKI against EGFR activating and resistant T790M mutated non-small cell lung cancer (NSCLC), additional mutant-selective inhibitors with a higher efficacy, especially for brain metastases, with favorable toxicity profile are still needed. In this study, we investigated the antitumor efficacy of YH25448, an oral, mutant-selective, irreversible third-generation EGFR-TKI in preclinical models. EXPERIMENTAL DESIGN: Antitumor activity of YH25448 was investigated in vitro using mutant EGFR-expressing Ba/F3 cells and various lung cancer cell lines. In vivo antitumor efficacy, ability to penetrate the blood-brain barrier (BBB), and skin toxicity of YH25448 were examined and compared with those of osimertinib using cell lines and PDX model. RESULTS: Compared with osimertinib, YH25448 showed a higher selectivity and potency in kinase assay and mutant EGFR-expressing Ba/F3 cells. In various cell line models harboring EGFR activating and T790M mutation, YH25448 effectively inhibited EGFR downstream signaling pathways, leading to cellular apoptosis. When compared in vivo at equimolar concentrations, YH25448 produced significantly better tumor regression than osimertinib. Importantly, YH25448 induced profound tumor regression in brain metastasis model with excellent brain/plasma and tumor/brain area under the concentration-time curve value. YH25448 rarely suppressed the levels of p-EGFR in hair follicles, leading to less keratosis than osimertinib in animal model. The potent systemic and intracranial activity of YH25448 has been shown in an ongoing phase I/II clinical trial for advanced EGFR T790M mutated NSCLC (NCT03046992). CONCLUSIONS: Our findings suggest that YH25448 is a promising third-generation EGFR inhibitor, which may be more effective and better tolerated than the currently approved osimertinib.

Asunto(s)

Neoplasias Encefálicas/secundario , Carcinoma de Pulmón de Células no Pequeñas/genética , Carcinoma de Pulmón de Células no Pequeñas/patología , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patología , Inhibidores de Proteínas Quinasas/farmacología , Acrilamidas/química , Acrilamidas/farmacología , Acrilamidas/uso terapéutico , Adulto , Compuestos de Anilina/química , Compuestos de Anilina/farmacología , Compuestos de Anilina/uso terapéutico , Animales , Barrera Hematoencefálica/efectos de los fármacos , Barrera Hematoencefálica/metabolismo , Neoplasias Encefálicas/diagnóstico , Neoplasias Encefálicas/tratamiento farmacológico , Neoplasias Encefálicas/mortalidad , Carcinoma de Pulmón de Células no Pequeñas/diagnóstico por imagen , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Modelos Animales de Enfermedad , Resistencia a Antineoplásicos/genética , Receptores ErbB/antagonistas & inhibidores , Receptores ErbB/química , Receptores ErbB/genética , Humanos , Neoplasias Pulmonares/diagnóstico por imagen , Masculino , Ratones , Modelos Moleculares , Mutación , Inhibidores de Proteínas Quinasas/química , Inhibidores de Proteínas Quinasas/uso terapéutico , Relación Estructura-Actividad , Resultado del Tratamiento , Ensayos Antitumor por Modelo de Xenoinjerto

RESUMEN

Notoginseng Radix and Rehmanniae Radix Preparata have been widely used traditionally for treating inflammatory diseases. This research studies the therapeutic effects of YH23537, the extracts of Notoginseng Radix and Rehmanniae Radix Preparata, on pain and cartilage degeneration in an experimental osteoarthritis (OA) model. Male Wistar rats were inoculated intra-articularly with 3 mg of monosodium iodoacetate (MIA) in the right intra-articular. Four days later, the animals were administrated orally with YH23537 daily for 24 days. Tactile allodynia and weight bearing were measured. Macroscopic and microscopic observations for articular cartilage were performed at the end of the experiment. Protein expression in the joint was determined by immunohistochemistry. The effects of YH23537 on mRNA levels in chondrocytes stimulated with interleukin (IL)-1ß were analyzed using random polymerase chain reaction. OA induction was confirmed by significant decrease of paw withdrawal latency, paw withdrawal threshold, and weight bearing compared with the normal group at 3 days after MIA injection. The YH23537-treated groups displayed significant increases in pain thresholds and weight bearing throughout the observation period. The damage to articular cartilage was significantly lessened visually and histopathologically by YH23537 treatment. YH23537 suppressed the expression of metalloproteinase-3, nitrotyrosine, IL-1ß and IL-6 increased in OA joints. YH23537 upregulated tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-3 in IL-1ß-stimulated human OA chondrocytes. The protein levels of the NF-κBp65 and HIF-2α in the joint tissues were reduced by YH23537. YH23537 exerted antinociceptive effects and cartilage protective effects in experimental OA rats by suppressing oxidative injury, inflammatory mediators, and inducing anabolic factors. We suggest that YH23537 may have efficacy for treating OA in humans.

Asunto(s)

Antiinflamatorios/farmacología , Enfermedades de los Cartílagos/tratamiento farmacológico , Osteoartritis/tratamiento farmacológico , Dolor/tratamiento farmacológico , Panax , Extractos Vegetales/farmacología , Rehmannia , Administración Oral , Animales , Antiinflamatorios/efectos adversos , Antiinflamatorios/uso terapéutico , Enfermedades de los Cartílagos/inducido químicamente , Cartílago Articular/efectos de los fármacos , Modelos Animales de Enfermedad , Yodoacetatos , Masculino , Osteoartritis/inducido químicamente , Dimensión del Dolor , Fitoterapia , Extractos Vegetales/administración & dosificación , Extractos Vegetales/uso terapéutico , Ratas , Ratas Wistar

RESUMEN

BACKGROUND AND PURPOSE: 5-HT4 receptor agonists have been shown to be effective at treating various gastrointestinal tract disorders. However, a lack of selectivity against off-targets has been a limiting factor for their clinical use. EXPERIMENTAL APPROACH: The binding affinity and selectivity of YH12852 for human 5-HT4(a) receptor in CHO-K1 cells were evaluated using radioligand binding assays, and agonistic activity was assessed using a ß-lactamase reporter system. Contractile activity and propulsive motility were measured in the guinea pig isolated distal colon. Its prokinetic effect on the gastrointestinal tract was evaluated in guinea pigs, dogs and monkeys. Its tissue distribution was evaluated in rats. KEY RESULTS: YH12852 exhibited high affinity and potency for human recombinant 5-HT4(a) receptor with high selectivity over other 5-HT and non-5-HT receptors, ion channels, enzymes and transporters. YH12852 induced contractions and increased propulsive motility in guinea pig isolated colon. These effects were abolished by the 5-HT4 receptor antagonist GR113808. YH12852 increased defecation more effectively than prucalopride in guinea pigs and dogs and improved gastric emptying more effectively than mosapride in guinea pigs, dogs and monkeys. YH12852 was highly distributed to the gastrointestinal tract as the target organ. CONCLUSION AND IMPLICATIONS: The high in vitro potency and selectivity of YH12852 for 5-HT4 receptor translated into potent in vivo efficacy with good tolerability. YH12852 significantly improved both upper and lower bowel motility in the animal models tested and has the potential to address considerable unmet needs in patients with functional constipation, gastroparesis or both.

Asunto(s)

Colon/efectos de los fármacos , Motilidad Gastrointestinal/efectos de los fármacos , Intestino Delgado/efectos de los fármacos , Receptores de Serotonina 5-HT4/metabolismo , Agonistas del Receptor de Serotonina 5-HT4/metabolismo , Agonistas del Receptor de Serotonina 5-HT4/farmacología , Animales , Células CHO , Colon/fisiología , Cricetinae , Cricetulus , Perros , Relación Dosis-Respuesta a Droga , Motilidad Gastrointestinal/fisiología , Humanos , Intestino Delgado/fisiología , Macaca fascicularis , Masculino , Pirimidinas , Ratas , Ratas Sprague-Dawley

RESUMEN

Resistin-like molecule alpha (Retnla), also known as 'Found in inflammatory zone 1', is a secreted protein that has been found in bronchoalveolar lavage (BAL) fluid of ovalbumin (OVA)-induced asthmatic mice and plays a role as a regulator of T helper (Th)2-driven inflammation. However, the role of Retnla in the progress of Th2-driven airway inflammation is not yet clear. To better understand the function of Retnla in Th2-driven airway inflammation, we generated Retnla-overexpressing (Retnla-Tg) mice. Retnla-Tg mice showed increased expression of Retnla protein in BAL fluid and airway epithelial cells. Retnla overexpression itself did not induce any alteration in lung histology or lung function compared to non-Tg controls. However, OVA-sensitized/challenged Retnla-Tg mice had decreased numbers of cells in BAL and inflammatory cells accumulating in the lung. They also showed a reduction in mucus production in the airway epithelium, concomitant with a decreased Muc5ac level. These results were accompanied by reduced levels of Th2 cytokines, including interleukin (IL)-4, IL-5, and IL-13, with no effect on levels of OVA-specific immunoglobulin isotypes. Furthermore, phosphorylation of ERK was markedly reduced in the lungs of OVA-challenged Retnla-Tg mice. Taken together, these results indicates that Retnla protects against Th2-mediated inflammation in an experimental mouse model of asthma, suggesting that therapeutic approaches to enhance the production of Retnla or Retnla-like molecules could be valuable for preventing allergic lung inflammation.

Asunto(s)

Asma/metabolismo , Hipersensibilidad/metabolismo , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Animales , Asma/genética , Secuencia de Bases , Líquido del Lavado Bronquioalveolar , Cartilla de ADN , Ensayo de Inmunoadsorción Enzimática , Hipersensibilidad/genética , Péptidos y Proteínas de Señalización Intercelular/genética , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Reacción en Cadena en Tiempo Real de la Polimerasa

RESUMEN

Gynecologic cancers constitute the fourth most common cancer type in women. Treatment outcomes are dictated by a multitude of factors, including stage at diagnosis, tissue type, and overall health of the patient. Current therapeutic options include surgery, radiotherapy, and chemotherapy, although significant unmet medical needs remain in regard to side effects and long-term survival. The efficacy of chemotherapy is influenced by cellular events such as the overexpression of oncogenes and downregulation of tumor suppressors, which together determine apoptotic responses. Phytochemicals are a broad class of natural compounds derived from plants, a number of which exhibit useful bioactive effects toward these pathways. High-throughput screening methods, rational modification, and developments in regulatory policies will accelerate the development of novel therapeutics based on these compounds, which will likely improve overall survival and quality of life for patients.

Asunto(s)

Resistencia a Antineoplásicos/efectos de los fármacos , Neoplasias de los Genitales Femeninos/tratamiento farmacológico , Fitoquímicos/administración & dosificación , Femenino , Neoplasias de los Genitales Femeninos/patología , Ensayos Analíticos de Alto Rendimiento , Humanos , Fitoquímicos/clasificación , Calidad de Vida , Resultado del Tratamiento

RESUMEN

AIMS: The aim of this study was to identify new compounds that induce cardiomyocyte differentiation of stem cells through cell-based screening and investigate lineage specificity and mechanisms in vitro. MAIN METHODS: Embryoid bodies (EBs) formed from TC-1/KH2 mouse embryonic stem cells (ESCs) carrying the gene for enhanced green fluorescent protein (EGFP) under the control of the α-myosin heavy chain (MHC) promoter were treated with test compounds. The number of cardiomyocyte-like (EGFP-expressing) cells in EBs was determined by fluorescence-activated cell sorting. Cardiomyocyte differentiation was further confirmed using lineage-specific biochemical assays and by investigating the expression of cardiomyocyte-specific and "stemness"-associated genes. Nuclear factor-kappaB (NF-κB) signaling activity was measured in A549 cells using a reporter-gene assay. KEY FINDINGS: A ß-carboline compound, designated CW108F, increased the number of mouse ESCs expressing α-MHC promoter-driven EGFP and the proportion of beating EBs. CW108F also increased expression of MHC in P19 stem cells, but did not induce osteogenesis of MC3T3-E1 cells, suggesting lineage-specific activity toward cardiomyocytes. CW108F upregulated expression of cardiac-specific GATA-4 and atrial natriuretic factor (ANF) genes in TC-1/KH2 cells, but downregulated expression of the stemness genes, Oct-4 and brachyury. CW108F inhibited NF-κB transcriptional activity, an effect that might contribute to its cardiomyogenesis-promoting activity. SIGNIFICANCE: The results of this study suggest that the novel ß-carboline, CW108F, promotes the differentiation of ESCs into cardiomyocytes and may be useful for investigating molecular pathways of cardiomyogenesis and generating cardiomyocytes from ESCs.

Asunto(s)

Carbolinas/farmacología , Diferenciación Celular/efectos de los fármacos , Cuerpos Embrioides/efectos de los fármacos , Células Madre Embrionarias/efectos de los fármacos , Miocitos Cardíacos/efectos de los fármacos , Células 3T3 , Animales , Cuerpos Embrioides/citología , Células Madre Embrionarias/citología , Regulación de la Expresión Génica/efectos de los fármacos , Proteínas Fluorescentes Verdes/genética , Ratones , Ratones Endogámicos C57BL , Miocitos Cardíacos/metabolismo , Cadenas Pesadas de Miosina/genética , FN-kappa B/metabolismo , Regiones Promotoras Genéticas

Asunto(s)

Hiperqueratosis Epidermolítica/genética , Ictiosis Ampollosa de Siemens/diagnóstico , Queratina-1/genética , Mutación , Adolescente , Análisis Mutacional de ADN , Diagnóstico Diferencial , Predisposición Genética a la Enfermedad , Humanos , Hiperqueratosis Epidermolítica/patología , Ictiosis Ampollosa de Siemens/genética , Ictiosis Ampollosa de Siemens/patología , Masculino , Fenotipo , Valor Predictivo de las Pruebas , Estructura Terciaria de Proteína , Índice de Severidad de la Enfermedad , Piel/patología

Asunto(s)

Colágeno Tipo VII/genética , Epidermólisis Ampollosa Distrófica/genética , Eliminación de Secuencia/genética , Epidermólisis Ampollosa Distrófica/patología , Femenino , Humanos , Recién Nacido , Reacción en Cadena de la Polimerasa

RESUMEN

Embryonic stem (ES) cells may be used as an alternative source of functionally intact cardiomyocytes for ischemic heart disease. Several natural and synthetic small molecules have been identified as useful tools for controlling and manipulating stem cell renewal and differentiation. Currently, there is an urgent requirement for novel small molecules that specifically induce differentiation of stem cells into cardiomyocytes. To identify compounds that promote cardiomyogenesis of stem cells, cell-based screening of a peptidomimetic small-molecule library was carried out. A series of ß-turn peptidomimetic compounds, including CW209E, increased the expression of α-MHC promoter-driven enhanced green fluorescent protein (EGFP) and ratio of beating embryoid bodies (EBs) without inducing cytotoxicity in mouse embryonic stem cells. CW209E also increased the number of beating EBs in human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs). Thus, this chemical compound should be useful for elucidation of the molecular pathway of cardiogenesis and generation of cardiomyocytes ex vivo, which can be further applied for experimental or clinical cell therapy for ischemic heart diseases.

Asunto(s)

Células Madre Embrionarias/efectos de los fármacos , Células Madre Pluripotentes Inducidas/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Peptidomiméticos/farmacología , Animales , Diferenciación Celular/efectos de los fármacos , Cuerpos Embrioides/metabolismo , Células Madre Embrionarias/metabolismo , Proteínas Fluorescentes Verdes/genética , Compuestos Heterocíclicos de 4 o más Anillos/farmacología , Compuestos Heterocíclicos de 4 o más Anillos/toxicidad , Ensayos Analíticos de Alto Rendimiento , Humanos , Hidrocarburos Fluorados/farmacología , Hidrocarburos Fluorados/toxicidad , Células Madre Pluripotentes Inducidas/metabolismo , Ratones , Ratones Endogámicos C57BL , Cadenas Pesadas de Miosina/genética , Peptidomiméticos/toxicidad , Regiones Promotoras Genéticas , Bibliotecas de Moléculas Pequeñas

RESUMEN

ETHNOPHARMACOLOGICAL RELEVANCE: Curcumin, a polyphenol compound from Curcuma longa L. has been used for centuries as an anti-inflammatory remedy including asthma. Curcumin has been reported to exert an anti-inflammatory effect, in part, through inhibition of the NF-κB pathway. AIM OF THE STUDY: The purposes of this study were to determine whether curcumin inhibits NF-κB-dependent transcription in vitro, and test whether treatment with curcumin reduces allergen-induced airway inflammation and hyper-responsiveness in a mouse model of asthma through inhibition of NF-κB pathway. MATERIALS AND METHODS: The effect of curcumin on NF-κB transcriptional activity was investigated using a cell-based luciferase reporter assay in A549 cells and by measuring inhibitory κBα (IκBα), p65, and p50 levels after exposure of Raw264.7 cells to lipopolysaccharide (LPS). BALB/c mice were sensitized to ovalbumin (OVA) by intraperitoneal injection, and challenged with repeated exposure to aerosolized OVA. The effects of daily administered curcumin (200mg/kg body weight, i.p.) on airway hyper-responsiveness (AHR), inflammatory cell number, and IgE levels in bronchoalveolar lavage (BAL) fluid were analyzed. NF-κB activation in lung tissue was also assessed by Western blot analyses. RESULTS: Curcumin inhibited NF-κB-dependent transcription in reporter assays in A549 cells with an IC(50) of 21.50±1.25µM. Curcumin stabilized IκBα and inhibited nuclear translocation of p65 and p50 in LPS-activated Raw264.7 cells, and curcumin-treated mice showed reduced nuclear translocation of p65 in lung tissue. Treatment with curcumin significantly attenuated AHR and reduced the numbers of total leukocytes and eosinophils in BAL fluid. Infiltration of inflammatory cells and mucus occlusions in lung tissue were significantly ameliorated by treatment with curcumin, which also markedly decreased the level of IgE in BAL fluid. CONCLUSION: Curcumin attenuates the development of allergic airway inflammation and hyper-responsiveness, possibly through inhibition of NF-κB activation in the asthmatic lung tissue. Our results indicate that curcumin may attenuate development of asthma by inhibition of NF-κB activation.

Asunto(s)

Antiinflamatorios/uso terapéutico , Asma/tratamiento farmacológico , Curcuma/química , Curcumina/uso terapéutico , FN-kappa B/antagonistas & inhibidores , Fitoterapia , Extractos Vegetales/uso terapéutico , Animales , Antiinflamatorios/farmacología , Asma/inmunología , Asma/metabolismo , Transporte Biológico/efectos de los fármacos , Líquido del Lavado Bronquioalveolar , Línea Celular , Curcumina/farmacología , Eosinófilos/metabolismo , Femenino , Proteínas I-kappa B/metabolismo , Inmunoglobulina E/metabolismo , Inflamación/tratamiento farmacológico , Inflamación/inmunología , Inflamación/metabolismo , Concentración 50 Inhibidora , Leucocitos/metabolismo , Pulmón/efectos de los fármacos , Pulmón/metabolismo , Ratones , Ratones Endogámicos BALB C , Moco/metabolismo , Inhibidor NF-kappaB alfa , Extractos Vegetales/farmacología , Rizoma , Factor de Transcripción ReIA/metabolismo

RESUMEN

BACKGROUNDS: Epidermolysis bullosa simplex (EBS) is a group of hereditary bullous disorders caused by mutations in the keratin genes KRT5 and KRT14. A significant genotype-phenotype correlation has been noted in previous studies of EBS. OBJECTIVE: In order to identify additional EBS mutations and elucidate the genotype-phenotype correlations in Korean EBS patients, we performed the first large scale mutational analysis of EBS patients of Korean origin. METHODS: We investigated fifteen Korean EBS patients by performing a sequence analysis of the entire coding sequences of KRT5 and KRT14. RESULTS: We identified six novel mutations, four within KRT5 (p.V143F, p.R265P, p.C479X and p.Asn177del), and two within KRT14 (p.R125L and p.L401P). In all, 13 missense, 1 nonsense, and 1 small deletion mutation were found. Five mutations in Dowling-Meara type (K14-p.R125H, K14-p.R125L, K5-E477K, K5-p.C479X and K5-p.Asn177del) were located in the highly conserved ends of the alpha-helical rod domain, the helix initiation (HIP), or helix termination (HTP) peptides of KRT5 and KRT14. Further, seven and three mutations were identified in EBS-generalized type and EBS-localized type, respectively. The positions of the mutations in both subtypes were more widely distributed within the rod domains and in the L12 linker domains of both keratin genes. CONCLUSIONS: This study should provide useful data and enhance our understanding of the EBS genotype-phenotype relationship. The genotype-phenotype correlation in Korean EBS patients was similar to previous studies performed in other ethnic groups. Lastly, our results confirmed that the mutational location in KRT5 or KRT14 is the most important factor in determining the phenotype severity.

Asunto(s)

Pueblo Asiatico/genética , Epidermólisis Ampollosa Simple/etnología , Epidermólisis Ampollosa Simple/genética , Queratina-14/genética , Queratina-5/genética , Mutación , Adolescente , Adulto , Codón sin Sentido , Epidermólisis Ampollosa Simple/patología , Femenino , Eliminación de Gen , Genotipo , Humanos , Lactante , Recién Nacido , Masculino , Persona de Mediana Edad , Mutación Missense , Fenotipo , Análisis de Secuencia , Índice de Severidad de la Enfermedad , Adulto Joven

RESUMEN

A series of methylenedioxybenzene compounds were synthesized and found to have hepatoprotective effects in chemical-induced hepatotoxicity models. The purpose of the present study was to investigate the anti-fibrotic effects of a synthetic methylenedioxybenzene compound, CW209292, using the dimethylnitrosamine (DMN)-induced chronic liver injury model in rats. Liver injuries were induced in Sprague Dawley rats by injection of DMN (intraperitoneally, 10 microl/kg) 3 times per week for 4 weeks. The rats were treated with CW209292 (per os, 25 or 75 mg/kg/d) for 4 weeks. Treatment of rats with DMN for 4 weeks resulted in significant decreases in serum albumin levels, whereas concomitant treatment with CW209292 prevented these decreases. CW209292 treatment also shortened prothrombin time prolonged by DMN, providing evidence that the agent was active in preserving liver function against DMN insult. DMN treatment caused marked increases in plasma bilirubin, aspartate aminotransferase (AST), alanine transaminase (ALT), and hyaluronic acid levels; CW209292 treatment reversed these increases. CW209292 also significantly reduced hepatic hydroxyproline content as well as hepatic fibrosis and inflammation in histological examination. Additionally, immunochemically detectable hepatic collagen type IV and alpha-smooth muscle actin levels were decreased by CW209292 treatment. Proliferation of hepatic stellate cells isolated from DMN-treated rats was inhibited by CW209292. Furthermore, tumor growth factor (TGF)-beta1 mRNA expression was increased in DMN-treated rats, whereas CW209292 treatment prevented these increases. These results suggest that CW209292 exhibits anti-fibrotic effects in Sprague Dawley rats with DMN-induced hepatic fibrosis by blocking the mRNA expression of TGF-beta1 and subsequent inhibition of the proliferation of hepatic stellate cells.

Asunto(s)

Benzodioxoles/uso terapéutico , Dimetilnitrosamina/toxicidad , Cirrosis Hepática/prevención & control , Tiazoles/uso terapéutico , Animales , Benzodioxoles/química , Benzodioxoles/farmacología , Peso Corporal/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Modelos Animales de Enfermedad , Femenino , Células Estrelladas Hepáticas/efectos de los fármacos , Células Estrelladas Hepáticas/patología , Hidroxiprolina/metabolismo , Inmunohistoquímica , Hígado/efectos de los fármacos , Hígado/enzimología , Hígado/metabolismo , Hígado/patología , Cirrosis Hepática/sangre , Cirrosis Hepática/inducido químicamente , Cirrosis Hepática/patología , Masculino , Estructura Molecular , Tiempo de Protrombina , Ratas , Ratas Sprague-Dawley , Análisis de Supervivencia , Tiazoles/química , Tiazoles/farmacología

Asunto(s)

Cromosomas Humanos Par 1 , Codón sin Sentido , Dermatitis Atópica/genética , Ictiosis Vulgar/genética , Proteínas de Filamentos Intermediarios/genética , Adulto , Pueblo Asiatico/genética , Dermatitis Atópica/complicaciones , Proteínas Filagrina , Humanos , Ictiosis Vulgar/complicaciones , Masculino

RESUMEN

CW-270031 is a novel synthesized carbapenem antibiotic with a broad antimicrobial activity. Carbapenem antibiotics are well known for their nephrotoxicity. In this study, we evaluated the nephrotoxicity potential of this compound in rabbits, which are known for being more sensitive than other animals to renal insult. CW-270031 was administered to NZW male rabbits via an ear vein (200 mg/kg, single injection). Blood samples were collected on 2, 3, and 4 days after treatment. Urea nitrogen and creatinine in plasma were quantified. Four days after the treatment, all animals were autopsied and histopathological examinations were performed on their kidneys, revealing that cephaloridine and imipenem were highly nephrotoxic, and cefazolin had mild renal toxicity, whereas CW-270031 as well as meropenem and tienam had no toxicity to the kidney. The present findings suggest that CW-270031 is a potential carbapenem antibiotic with no nephrotoxicity.

Asunto(s)

Antibacterianos/efectos adversos , Carbapenémicos/efectos adversos , Enfermedades Renales/inducido químicamente , Riñón/efectos de los fármacos , Pirrolidinas/efectos adversos , Animales , Antibacterianos/administración & dosificación , Antibacterianos/química , Carbapenémicos/administración & dosificación , Carbapenémicos/química , Cilastatina/administración & dosificación , Cilastatina/efectos adversos , Cilastatina/química , Imipenem/administración & dosificación , Imipenem/efectos adversos , Imipenem/química , Riñón/patología , Masculino , Meropenem , Pirrolidinas/administración & dosificación , Pirrolidinas/química , Conejos , Tienamicinas/administración & dosificación , Tienamicinas/efectos adversos , Tienamicinas/química

RESUMEN

3'-Phosphoinositide-dependent kinase-1 (PDK1) has been identified for its ability to phosphorylate and activate Akt. Accumulated studies have shown that the activation of the PDK1/Akt pathway plays a pivotal role in cell survival, proliferation, and tumorigenesis. Therefore, the PDK1/Akt pathway is believed to be a critical target for cancer intervention. In this paper, we report the discovery of a new function of phenothiazines, widely known as antipsychotics, inhibiting PDK1/Akt pathway. Upon epidermal growth factor (EGF) stimulation, phenothiazines specifically suppressed the kinase activity of PDK1 and the phosphorylation level of Akt. The inhibition of PDK1/Akt kinase resulted in suppression of EGF-induced cell growth and induction of apoptosis in human ovary cancer cells. In particular, phenothiazines were highly selective for downstream targets of PDK1/Akt and did not inhibit the activation of phosphatidylinositol 3-kinase (PI3K), EGFR, or extracellular signal-regulated kinase 1/2 (ERK1/2). In particular, phenothiazines effectively suppressed tumor growth in nude mice of human cancer cells. Taken together, these findings provide strong evidence for novel function of phenothiazines, pharmacologically targeting PDK1/Akt for anticancer drug discovery.

Asunto(s)

Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Fenotiazinas/farmacología , Proteínas Serina-Treonina Quinasas/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-akt/antagonistas & inhibidores , Transducción de Señal , Animales , Humanos , Ratones , Fosforilación , Piruvato Deshidrogenasa Quinasa Acetil-Transferidora , Trasplante Heterólogo

Asunto(s)

Epidermólisis Ampollosa Simple/genética , Queratina-5/genética , Fenotipo , Adulto , Femenino , Genotipo , Humanos , Lactante , Queratina-5/metabolismo , Masculino , Piel/metabolismo , Piel/patología

RESUMEN

CW-270033, an injectable carbapenem, is a novel, synthesized pyrrolidinyl-thio carbapenem. In the present study, the in-vitro and in-vivo antibacterial activities of CW-270033 against wild-type strains and clinical isolates were compared with those of imipenem and meropenem. CW-270033 was more active than imipenem against Gram-negative (Escherichia coli and Pseudomonas aeruginosa) clinical isolates, but was slightly less active than meropenem. Against the Gram-positive clinical isolates methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus (VRE), CW-270033 was slightly more active than meropenem, but was less active than imipenem. CW-270033 displayed potent in-vivo activity against E. coli ATCC 25922, P. aeruginosa ATCC 27853, and S. aureus SMITH in the mouse systemic infection model; the efficacy of CW-270033 in this model was 2--7 fold higher than that of meropenem. This activity was comparable to the in-vitro activity of CW-270033. An intravenous injection of CW-270033 showed that the half-life of CW-270033 in serum in mice was about 20 min, which was about two times that of meropenem. CW-270033 was also found to be resistant to hydrolysis by the mouse renal dehydropeptidase I (DHP-I) enzyme.

Asunto(s)

Antibacterianos/farmacología , Bacterias/efectos de los fármacos , Infecciones Bacterianas/prevención & control , Carbapenémicos/farmacología , Pirrolidinas/farmacología , Animales , Antibacterianos/administración & dosificación , Antibacterianos/química , Bacterias/crecimiento & desarrollo , Infecciones Bacterianas/microbiología , Carbapenémicos/administración & dosificación , Carbapenémicos/química , Cilastatina/administración & dosificación , Cilastatina/química , Cilastatina/farmacología , Relación Dosis-Respuesta a Droga , Enterococcus faecalis/efectos de los fármacos , Enterococcus faecalis/crecimiento & desarrollo , Escherichia coli/efectos de los fármacos , Escherichia coli/crecimiento & desarrollo , Inyecciones Intravenosas , Meropenem , Resistencia a la Meticilina , Ratones , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Inhibidores de Proteasas/administración & dosificación , Inhibidores de Proteasas/química , Inhibidores de Proteasas/farmacología , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/crecimiento & desarrollo , Pirrolidinas/administración & dosificación , Pirrolidinas/química , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/crecimiento & desarrollo , Tienamicinas/administración & dosificación , Tienamicinas/farmacología , Resistencia a la Vancomicina

RESUMEN

OBJECTIVE: To evaluate whether serum replacement with growth factor-free synthetic substances contributed to the effective establishment of embryonic stem (ES) cells. DESIGN: Randomized, prospective model study. SETTING: Gamete and stem cell biotechnology laboratory at Seoul National University in Korea. ANIMAL(S): F1 (C57BL6 x DBA2) mice. INTERVENTION(S): Blastocysts of different origins were cultured in serum-replaced media. MAIN OUTCOME MEASURE(S): Embryonic stem cell establishment. RESULT(S): Eight batches of ES cells were established from colony-forming inner cell mass cells after the replacement of fetal bovine serum (FBS) with synthetic knockout serum replacement (KSR) in mkDMEM. The established cells were positive for ES cell markers and formed both embryoid bodies in vitro and teratomas in vivo, but the established cell batches and control (transformed) ES cells responded differently to the culture media. Higher levels of cell viability were detected after the replacement with the 75:25 FBS-KSR mixture than with any other mixtures, and a gradual decrease in viability was detected as the KSR volume ratio was increased. The 75:25 FBS-KSR mixture-containing medium supported ES cell establishment of outbred ICR, F1, and F2 of C57BL6/DBA2; F1 parthenogenetic and ES cell-complemented tetraploid blastocysts; and single ES-cell cultures. CONCLUSION(S): A serum-replaced medium could be used for effective ES-cell establishment of various origins.

Asunto(s)

Medios de Cultivo/síntesis química , Medios de Cultivo/farmacología , Embrión de Mamíferos/citología , Células Madre/citología , Células Madre/efectos de los fármacos , Animales , Animales no Consanguíneos , Biomarcadores/metabolismo , Blastocisto/citología , Blastocisto/efectos de los fármacos , Bovinos/sangre , Supervivencia Celular , Células Cultivadas , Medios de Cultivo/química , Femenino , Sangre Fetal , Sustancias de Crecimiento/análisis , Masculino , Ratones , Ratones Endogámicos , Células Madre/metabolismo , Células Madre/fisiología