RESUMEN
The currently used anti-cytokine therapeutic antibodies cannot selectively neutralize pathogenic cytokine signalling that cause collateral damage to protective signalling cascades. The single domain chain firstly discovered in Camelidae displays fully functional ability in antigen-binding against variable targets, which has been seemed as attractive candidates for the next-generation biologic drug study. In this study, we established a simple prokaryotic expression system for a dual target-directed single domain-based fusion protein against the interleukin-6 receptor and human serum, albumin, the recombinant anti-IL-6R fusion protein (VHH-0031). VHH-0031 exhibited potent anti-inflammatory effects produced by LPS on cell RAW264.7, where the major cytokines and NO production were downregulated after 24 h incubation with VHH-0031 in a dose-dependent manner. In vivo, VHH-0031 presented significant effects on the degree reduction of joint swelling in the adjuvant-induced arthritis (AIA) rat, having a healthier appearance compared with the dexamethasone. The expression level of JNK protein in the VHH-0031 group was significantly decreased, demonstrating that VHH-0031 provides a low-cost and desirable effect in the treatment of more widely patients.
Asunto(s)
Antiinflamatorios/inmunología , Artritis Experimental/tratamiento farmacológico , Interleucina-6/antagonistas & inhibidores , Albúmina Sérica Humana/antagonistas & inhibidores , Anticuerpos de Dominio Único/inmunología , Animales , Antiinflamatorios/uso terapéutico , Especificidad de Anticuerpos , Artritis Experimental/inmunología , Citocinas/metabolismo , ADN Complementario/genética , Dexametasona/uso terapéutico , Evaluación Preclínica de Medicamentos , Inducción Enzimática/efectos de los fármacos , Humanos , Interleucina-6/inmunología , Lipopolisacáridos/toxicidad , MAP Quinasa Quinasa 4/biosíntesis , MAP Quinasa Quinasa 4/genética , Ratones , Modelos Moleculares , Terapia Molecular Dirigida , Óxido Nítrico/metabolismo , Conformación Proteica , Células RAW 264.7 , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/inmunología , Albúmina Sérica Humana/inmunología , Anticuerpos de Dominio Único/genéticaRESUMEN
OBJECTIVE: To investigate the chemical constituents from the stems and branches of Sorbaria arborea. METHODS: The chemical constituents were isolated and purified by silica gel column chromatography, Sephadex LH-20 column chromatography and recrystallization. Their structures were identified by physicochemical properties and spectra analysis. RESULTS: Ten compounds were isolated and identified as ursolic acid (1), cucurbitacin F (2), (-) -epicatechin (3), daucosterol (4), arbutin (5), 3-O-ß-anthemisol (6), 2,6-dimethoxy-p-hydroquinone-4-O-ß-D-glucopyranoside (7), lupeol (8), betulin (9) and lup-20 (29) -en-3ß, 30-diol (10). CONCLUSION: All the compounds are isolated from this plant for the first time, and compounds 1, 6 - 8 and 10 are obtained from Sorbaria genus for the first time.