RESUMEN
In this study, a novel hydrogel system incorporating an amino acid-based deep eutectic solvent (DES) was prepared, and the skin-permeation enhancement of traditional Chinese herb medicine was evaluated using "sanwujiaowan" extract as the model formula. Briefly, a DES-extract complex was constructed by co-heating the herb formula extracts with the amino acid as the hydrogen receptor and citric acid as the hydrogen donor. The DES-extract complex demonstrated excellent dissolution and skin permeability of the complicated ingredients in the extracts. Consequently, the DES-extract complex was introduced to a hydrogel system, which showed better mechanical properties and viscoelasticity performance. Using a collagen-induced arthritis rat model, the DES-hydrogels exerted an enhanced therapeutic effect that significantly reduced the inflammatory response with systemic toxicity of the extracts. Therefore, our work suggests a novel strategy for synergistic transdermal delivery of Chinese herb medicine and local treatments for rheumatoid arthritis.
Asunto(s)
Artritis Reumatoide , Hidrogeles , Aminoácidos , Animales , Artritis Reumatoide/tratamiento farmacológico , China , Ácido Cítrico , Disolventes Eutécticos Profundos , Hidrogeles/química , Hidrógeno , RatasRESUMEN
Combinatory enhancement of innate and adaptive immune responses is a promising strategy in immunotherapeutic drug development. Bifunctional macromolecules that simultaneously target two mechanisms may provide additional advantages over the combination of targeting two single pathways. Interferon alpha (IFNα) has been used clinically against viral infection such as the chronic infection of hepatitis B virus (CHB) as well as some types of cancers. OX40 is a costimulatory immune checkpoint molecule involved in the activation of T lymphocytes. To test whether simultaneously activating IFNα and OX40 signaling pathway could produce a synergistic therapeutic effect on CHB and tumors, we designed a bifunctional fusion protein composed of a mouse OX40 agonistic monoclonal antibody (OX86) and a mouse IFNα4, joined by a flexible (GGGGS)3 linker. This fusion protein, termed OX86-IFN, can activate both IFNα and OX40. We demonstrated that OX86-IFN could effectively activate T lymphocytes in the peripheral blood of mice. Furthermore, we showed that OX86-IFN had superior therapeutic effect to monotherapies in HBV hydrodynamic transfection and syngeneic tumor models. Collectively, our data suggests that simultaneously targeting interferon and OX40 signaling pathways by bifunctional molecule OX86-IFN elicits potent antiviral and antitumor activities, which could provide a new strategy in developing therapeutic agents against viral infection and tumors.
Asunto(s)
Antineoplásicos Inmunológicos/farmacología , Antivirales/farmacología , Hepatitis B Crónica/tratamiento farmacológico , Interferón-alfa/agonistas , Neoplasias/tratamiento farmacológico , Receptores OX40/agonistas , Proteínas Recombinantes de Fusión/farmacología , Inmunidad Adaptativa/efectos de los fármacos , Animales , Antineoplásicos Inmunológicos/administración & dosificación , Antivirales/administración & dosificación , Linfocitos T CD4-Positivos/efectos de los fármacos , Linfocitos T CD4-Positivos/metabolismo , Linfocitos T CD8-positivos/efectos de los fármacos , Linfocitos T CD8-positivos/metabolismo , Línea Celular Tumoral , Modelos Animales de Enfermedad , Virus de la Hepatitis B/efectos de los fármacos , Inmunidad Innata/efectos de los fármacos , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Terapia Molecular Dirigida/métodos , Monocitos/efectos de los fármacos , Monocitos/metabolismo , Trasplante de Neoplasias , Receptor de Interferón alfa y beta/agonistas , Proteínas Recombinantes de Fusión/administración & dosificaciónRESUMEN
Both raw and vinegar products of the rhizome of Curcuma phaeocaulis are common drugs for promoting blood circulation and removing blood stasis in traditional Chinese medicine,which could be reflected in the inhibition of tail thrombosis in mice. As the traditional processing theory instructs,vinegar tastes sour and bitter,but can activate blood circulation and remove stasis after being infiltrated into the rhizome of C. phaeocaulis as an excipient. In this study,under the help of the ultrafast liquid chromatography-quadrupole time-offlight mass spectrometry( UFLC-Q-TOF-MS),the spectrum-effect relationship between the inhibition of tail thrombosis in mice and the rhizome of C. phaeocaulis both before and after the vinegar processing,were established to explore the functional changes of blood circulation and stasis after vinegar process. Based on the peak area from the fingerprint of UFLC-Q-TOF-MS of the alcohol extracts from the raw and vinegar-processed rhizome of C. phaeocaulis and their efficacy for inhibiting tail thrombosis,the correlation between the chromatography of UFLC-Q-TOF-MS and the inhibition of tail thrombosis in mice were analyzed by orthogonal partial least squares discriminant analysis( OPLS-DA) method. The results,produced by Simca-P software,showed that effective components consisted of eight peaks 16,24( aromadendrene oxide),3,11,22( dehydro-α-curcumene),19[( R)-(-)-α-curcumene],23 and 10 from the fingerprint,making great contribution to distinguish C. phaeocaulis raw products and the corresponding vinegar processed products. Therefore,from the perspective of inhibiting the formation of tail thrombosis in mice,the marker components could be found through the spectrum-effect relationship to distinguish C.phaeocaulis raw and vinegar products. This study provided new basis to explain the difference between the raw and the processed products of traditional Chinese medicine in the functional change of promoting blood circulation and removing blood stasis.
Asunto(s)
Curcuma/química , Medicamentos Herbarios Chinos/farmacología , Trombosis/tratamiento farmacológico , Ácido Acético , Animales , Cromatografía Líquida de Alta Presión , Medicamentos Herbarios Chinos/química , Espectrometría de Masas , Ratones , Rizoma/químicaRESUMEN
Excessive activation of hepatic stellate cells (HSCs) is a key step in liver fibrogenesis. Here we report that CUG-binding protein 1 (CUGBP1) expression is elevated in HSCs and positively correlates with liver fibrosis severity in human liver biopsies. Transforming growth factor-beta (TGF-ß) selectively increases CUGBP1 expression in cultured HSCs in a p38 mitogen-activated protein kinase (MAPK)-dependent manner. Knockdown of CUGBP1 inhibits alpha smooth muscle actin (α-SMA) expression and promotes interferon gamma (IFN-γ) production in HSCs in vitro. We further show that CUGBP1 specifically binds to the 3' untranslated region (UTR) of human IFN-γ mRNA and promotes its decay. In mice, knockdown of CUGBP1 alleviates, whereas its overexpression exacerbates, bile duct ligation (BDL)-induced hepatic fibrosis. Therefore, CUGBP1-mediated IFN-γ mRNA decay is a key event for profibrotic TGF-ß-dependent activation of HSCs, and inhibiting CUGBP1 to promote IFN-γ signalling in activated HSCs could be a novel strategy to treat liver fibrosis.
Asunto(s)
Proteínas CELF1/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Cirrosis Hepática/inducido químicamente , Regiones no Traducidas 3' , Animales , Benzofuranos/farmacología , Proteínas CELF1/genética , Células Cultivadas , Regulación de la Expresión Génica/fisiología , Hepatocitos/metabolismo , Humanos , Interferón gamma , Cirrosis Hepática/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Unión Proteica , Interferencia de ARN , ARN Mensajero/genética , ARN Mensajero/metabolismo , Factor de Crecimiento Transformador beta/genética , Factor de Crecimiento Transformador beta/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/genética , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
A series of novel hedgehog signaling pathway inhibitors have been designed by structural modification based on the former reported scaffold of 4-(2-pyrimidinylamino)benzamide. The SAR for this series was described and many derivatives showed potent inhibitory activity. Among these compounds, compounds 12af and 12bf were identified to have high potency and optimal PK profiles. Although both of compounds 12af and 12bf did not show strong antitumor efficacy in LS-174T nude mice model, they were promising candidates as Hh signaling inhibitors due to great potency against Hh signaling pathway and outstanding PK properties, deserving further evaluation in other Hh signaling operative tumor models.
Asunto(s)
Antineoplásicos/farmacología , Antineoplásicos/farmacocinética , Benzamidas/farmacología , Benzamidas/farmacocinética , Proteínas Hedgehog/antagonistas & inhibidores , Transducción de Señal/efectos de los fármacos , Administración Oral , Aminación , Animales , Antineoplásicos/administración & dosificación , Benzamidas/administración & dosificación , Línea Celular Tumoral , Proteínas Hedgehog/metabolismo , Humanos , Masculino , Ratones Endogámicos BALB C , Ratones Desnudos , Neoplasias/tratamiento farmacológico , Neoplasias/metabolismo , Piridinas/administración & dosificación , Piridinas/farmacocinética , Piridinas/farmacología , Relación Estructura-ActividadRESUMEN
Epigallocatechin-3-gallate (EGCG) exerts anti-inflammatory properties on immune cells and binds to CD4 molecules. However, the effects of EGCG on CD4(+) T cells remain largely unknown. Here, we found that EGCG enhanced IFN-γ-induced signal transducer and activator of transcription 1 (STAT1) activation in primary CD4(+) T cells from C57BL/6 mice and in a human leukemic CD4(+) T-cell line of Hut 78 cells, while it inhibited the classical pathway of IFN-γ signaling including activating phosphorylations of Janus kinase (JAK) 1, JAK2 and STAT3, forming interferon-γ activated sequence (GAS)-binding STAT1 homodimers, and producing pro-inflammatory chemokine (C-X-C motif) ligand 9 (CXCL9). CD4 blockade did not suppress the increase in IFN-γ-induced STAT1 activation in CD4(+) T cells by EGCG. Furthermore, activation of Src kinase was also triggered by IFN-γ plus EGCG in both Hut 78 and primary CD4(+) T cells. Interestingly, EGCG promoted apoptosis of CD4(+) T cells treated with IFN-γ. The increases in STAT1 activation and apoptosis induced by EGCG in IFN-γ-activated CD4(+) T cells were almost completely abolished by a selective Src family kinase inhibitor, SU6656. Moreover, EGCG alleviates CD4(+) CD45RB(hi) CD25(-) T cell transfer induced colitis with less accumulation of CD4(+) T cells in the colon. In conclusion, the present study reports an alternative activation of STAT1 via Src by EGCG in IFN-γ-activated CD4(+) T cells, which promotes the apoptosis of IFN-γ-activated CD4(+) T cells and contributes to the improvement of T cell-mediated colitis. Our findings suggest a novel role of EGCG in regulating IFN-γ signaling and controlling inflammation.
Asunto(s)
Apoptosis/efectos de los fármacos , Linfocitos T CD4-Positivos/efectos de los fármacos , Catequina/análogos & derivados , Interferón gamma/farmacología , Factor de Transcripción STAT1/metabolismo , Animales , Apoptosis/fisiología , Linfocitos T CD4-Positivos/fisiología , Catequina/farmacología , Línea Celular , Femenino , Humanos , Janus Quinasa 1/genética , Janus Quinasa 1/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones SCID , Receptores de Interferón/genética , Receptores de Interferón/metabolismo , Factor de Transcripción STAT1/genética , Transducción de Señal , Bazo/citología , Familia-src Quinasas/efectos de los fármacos , Familia-src Quinasas/metabolismo , Receptor de Interferón gammaRESUMEN
The side effects of current immunosuppressive drugs have impeded the development of therapies for immune diseases. Selective regulation of STAT signaling is an attractive strategy for treating immune disorders. In this study, we used a small-molecule compound to explore possible means of targeting STAT1 for the treatment of Th1-mediated inflammation. Selective regulation of STAT1 signaling in T cells from C57BL/6 mice was accomplished using fusaruside, a small-molecule compound that triggers the tyrosine phosphorylation of Src homology 2-containing protein tyrosine phosphatase 2 (SHP-2). The interaction of tyrosine phosphorylated SHP-2 (pY-SHP-2) with cytosolic STAT1 prevented the recruitment of STAT1 to IFN-γR and specifically inhibited STAT1 signaling, resulting in a reduction in Th1 cytokine production and an improvement in 2, 4, 6-trinitrobenzene sulfonic acid-induced colitis in mice. Blocking the pY-SHP-2-STAT1 interaction, with SHP-2 inhibitor NSC-87877 or using T cells from conditional SHP-2 knockout mice, reversed the effects of fusaruside, resulting in STAT1 activation and worsened colitis. The fusaruside-induced ability of pY-SHP-2 to selectively sequestrate STAT1 from recruitment to the receptor is independent of its function as a phosphatase, demonstrating a novel role for SHP-2 in regulating both STAT1 signaling and Th1-type immune responses. These findings could lead to increased options for the treatment of Crohn's disease and other Th1-mediated inflammatory diseases.
Asunto(s)
Colitis/inmunología , Colitis/terapia , Citosol/inmunología , Citosol/metabolismo , Inmunosupresores/farmacología , Mediadores de Inflamación/fisiología , Proteína Tirosina Fosfatasa no Receptora Tipo 11/metabolismo , Factor de Transcripción STAT1/metabolismo , Animales , Células Cultivadas , Cerebrósidos/metabolismo , Cerebrósidos/fisiología , Colitis/metabolismo , Modelos Animales de Enfermedad , Fusarium/química , Inmunosupresores/metabolismo , Mediadores de Inflamación/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones SCID , Ratones Transgénicos , Fosforilación/inmunología , Unión Proteica/inmunología , Transporte de Proteínas/inmunología , Proteína Tirosina Fosfatasa no Receptora Tipo 11/deficiencia , Proteína Tirosina Fosfatasa no Receptora Tipo 11/fisiología , Factor de Transcripción STAT1/antagonistas & inhibidores , Factor de Transcripción STAT1/fisiología , Transducción de Señal/inmunología , Células TH1/inmunología , Células TH1/metabolismo , Células TH1/patologíaRESUMEN
Erlotinib is a potent inhibitor of epidermal growth factor receptor tyrosine kinase and has been demonstrated to treat advanced or metastatic non-small cell lung cancer to prolong survival after failure of first-line or second-line chemotherapy. However, little is known about its effects on immune system. In the present study, we aimed to investigate the immunosuppressive activity of erlotinib on T lymphocytes both in vitro and in vivo, and further explore its potential molecular mechanism. Erlotinib exerted a significant inhibition on the T cell proliferation and activation induced by concanavalin A, anti-CD3 plus anti-CD28, staphylococcal enterotoxin B or phorbol myristate acetate respectively in a concentration-dependent manner and it also inhibited the secretion of the proinflammatory cytokines such as IL-2 and IFN-γ of activated T cells. Further study showed that erlotinib caused G0/G1 arrest and suppressed the phosphorylations of c-Raf, ERK and Akt in activated T cells. Moreover, erlotinib significantly ameliorated picryl chloride-induced ear contact dermatitis in a dose-dependent manner in vivo. In summary, these findings suggest that erlotinib may cause the impairment of T-cell-mediated immune response both in vitro and in vivo through inhibiting T cell proliferation and activation, which is closely associated with its potent down-regulation of the c-Raf/ERK cascade and Akt signaling pathway.
Asunto(s)
Regulación hacia Abajo/efectos de los fármacos , Quinasas MAP Reguladas por Señal Extracelular/antagonistas & inhibidores , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Proteína Oncogénica v-akt/antagonistas & inhibidores , Inhibidores de Proteínas Quinasas/toxicidad , Proteínas Proto-Oncogénicas c-raf/antagonistas & inhibidores , Quinazolinas/toxicidad , Subgrupos de Linfocitos T/efectos de los fármacos , Animales , Células Cultivadas , Relación Dosis-Respuesta a Droga , Regulación hacia Abajo/inmunología , Clorhidrato de Erlotinib , Quinasas MAP Reguladas por Señal Extracelular/fisiología , Femenino , Sistema de Señalización de MAP Quinasas/inmunología , Ratones , Ratones Endogámicos BALB C , Proteína Oncogénica v-akt/fisiología , Subgrupos de Linfocitos T/inmunologíaRESUMEN
In the present study, we aimed to investigate the immunosuppressive activity of jaceosidin, a flavone isolated from Artemisia vestita, on T lymphocytes both in vitro and in vivo, and further explore its potential molecular mechanism. Jaceosidin exerted a significant inhibition on the T cell proliferation and activation induced by concanavalin A (Con A) in a concentration-dependent manner and it also inhibited the secretion of the proinflammatory cytokines such as IL-2, TNF-α and IFN-γ of activated T cells. Further study showed that jaceosidin down-regulated STAT1 activation and T-bet expression in activated T cells. Moreover, in order to investigate the immunosuppressive effect of jaceosidin in vivo, the picryl chloride (PCl)-induced ear contact dermatitis model was performed on BALB/c mice. Jaceosidin significantly ameliorated PCl-induced ear swelling in a dose-dependent manner, which was due to its inhibition of the STAT1/T-bet signaling pathway. In summary, these findings suggest that jaceosidin exerts its immunosuppressive effect both in vitro and in vivo through inhibiting T cell proliferation and activation, which is closely associated with its potent down-regulation of the IFN-γ/STAT1/T-bet signaling pathway.