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Introduction: The antinociceptive and pharmacological activities of C-Phycocyanin (C-PC) and Phycocyanobilin (PCB) in the context of inflammatory arthritis remain unexplored so far. In the present study, we aimed to assess the protective actions of these compounds in an experimental mice model that replicates key aspects of human rheumatoid arthritis. Methods: Antigen-induced arthritis (AIA) was established by intradermal injection of methylated bovine serum albumin in C57BL/6 mice, and one hour before the antigen challenge, either C-PC (2, 4, or 8 mg/kg) or PCB (0.1 or 1 mg/kg) were administered intraperitoneally. Proteome profiling was also conducted on glutamate-exposed SH-SY5Y neuronal cells to evaluate the PCB impact on this key signaling pathway associated with nociceptive neuronal sensitization. Results and discussion: C-PC and PCB notably ameliorated hypernociception, synovial neutrophil infiltration, myeloperoxidase activity, and the periarticular cytokine concentration of IFN-γ, TNF-α, IL-17A, and IL-4 dose-dependently in AIA mice. In addition, 1 mg/kg PCB downregulated the gene expression for T-bet, RORγ, and IFN-γ in the popliteal lymph nodes, accompanied by a significant reduction in the pathological arthritic index of AIA mice. Noteworthy, neuronal proteome analysis revealed that PCB modulated biological processes such as pain, inflammation, and glutamatergic transmission, all of which are involved in arthritic pathology. Conclusions: These findings demonstrate the remarkable efficacy of PCB in alleviating the nociception and inflammation in the AIA mice model and shed new light on mechanisms underlying the PCB modulation of the neuronal proteome. This research work opens a new avenue to explore the translational potential of PCB in developing a therapeutic strategy for inflammation and pain in rheumatoid arthritis.

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Cytokines, demyelination and neuroaxonal degeneration in the central nervous system are pivotal elements implicated in the pathogenesis of multiple sclerosis (MS) and its nonclinical model of experimental autoimmune encephalomyelitis (EAE). Phycocyanobilin (PCB), a chromophore of the biliprotein C-Phycocyanin (C-PC) from Spirulina platensis, has antioxidant, immunoregulatory and anti-inflammatory effects in this disease, and it could complement the effect of other Disease Modifying Treatments (DMT), such as Interferon-ß (IFN-ß). Here, our main goal was to evaluate the potential PCB benefits and its mechanisms of action to counteract the chronic EAE in mice. MOG35-55-induced EAE was implemented in C57BL/6 female mice. Clinical signs, pro-inflammatory cytokines levels by ELISA, qPCR in the brain and immunohistochemistry using precursor/mature oligodendrocytes cells antibodies in the spinal cord, were assessed. PCB enhanced the neurological condition, and waned the brain concentrations of IL-17A and IL-6, pro-inflammatory cytokines, in a dose-dependent manner. A down- or up-regulating activity of PCB at 1 mg/kg was identified in the brain on three (LINGO1, NOTCH1, and TNF-α), and five genes (MAL, CXCL12, MOG, OLIG1, and NKX2-2), respectively. Interestingly, a reduction of demyelination, active microglia/macrophages density, and axonal damage was detected along with an increase in oligodendrocyte precursor cells and mature oligodendrocytes, when assessed the spinal cords of EAE mice that took up PCB. The studies in vitro in rodent encephalitogenic T cells and in vivo in the EAE mouse model with the PCB/IFN-ß combination, showed an enhanced positive effect of this combined therapy. Overall, these results demonstrate the anti-inflammatory activity and the protective properties of PCB on the myelin and support its use with IFN-ß as an improved DMT combination for MS.

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BACKGROUND: More potent antitumor activity is desired in Interferon (IFN)-treated cancer patients. This could be achieved by combining IFN alpha and IFN gamma. The aim of this work was to characterize the pharmacokinetics and pharmacodynamics of a novel formulation containing a co-formulated combination of IFNs alpha-2b and gamma (CIGB-128-A). METHODS: A group of nine healthy male subjects received intramuscularly 24.5 × 106 IU of CIGB-128-A. IFN concentrations were evaluated for 48 h. Serum neopterin, beta2-microglobulin (ß2M) and 2'-5' oligoadenylate synthetase (2'-5' OAS), classical IFN-inducible serum markers, were measured during 192 h by enzyme immunoassay and body temperature was used as pharmacodynamic variable as well. RESULTS: Concerning pharmacokinetics, serum IFNs' profiles were better fitted to a mono-compartmental model with consecutive zero order and first order absorption, one bioavailability value. No interferences by simultaneous administered IFNs were observed in their typical similar systemic profiles. Neopterin and ß2M time profiles showed a delay that was efficiently linked to pharmacokinetics by means of a zero order absorption rate constant. Neopterin level was nine-fold higher than initial values, 48 h post-administration, an increment not described before. At this time, mean serum ß2M peaked around the double from baseline. Serum concentrations of the enzyme 2'-5' OAS was still elevated on the 8 day post-injection. The formulation was well tolerated. Most frequent adverse reactions were fever, headache, arthralgia and lymphopenia, mostly mild. CONCLUSIONS: The administration of co-formulated IFN alpha-2b and IFN gamma likely provides improved pharmacodynamic properties that may be beneficial to treat several malignancies. TRIAL REGISTRATION: Cuban Public Registry of Clinical Trials RPCEC00000118 , May 24, 2011.

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La producción de medicamentos es hoy muy demandada debido al alto rigor que imponen las regulaciones y normativas actuales. Ese rigor no es solo aplicable a las facilidades de fabricación, sino que también es exigido en la fase de investigación y desarrollo. Nuestro país se destaca actualmente por una industria biotecnológica creciente que ha puesto ya en el mercado nacional e internacional importantes medicamentos de probada eficacia en el tratamiento de patologías como el cáncer. El Centro de Isótopos es una institución, cuyas instalaciones están a disposición de esa industria, brindando una plataforma de trabajo para llevar a cabo investigaciones que permitan dar respuesta a estudios de biodistribución y farmacocinética en modelos experimentales relevantes. Durante los años de experiencia en estas actividades se ha contribuido a la investigación y desarrollo de productos de diversa naturaleza como los biotecnológicos, experimentando una evolución hacia la puesta a punto de nuevas tecnologías y a la incorporación de metodologías más adecuadas a los estándares actuales. La aplicación del marcaje de moléculas con isótopos radiactivos ha demostrado su vigencia, especialmente en su conveniencia para obtener imágenes de distribución de los fármacos y su cinética. Se dispone de técnicas para dar respuesta a las más importantes investigaciones que se demanden.

Drugs production is a highly demanding industry because of the high rigor of current regulations and standards which apply to manufacturing facilities. They are also required in the research and development stage. Our national biotechnological industry is developing and producing important drugs for national and international market to treat diseases like cancer. The Isotope Centre is an institution supporting such a development by means of a work platform to carry out researches in the field of pharmacokinetic and biodistribution in experimental models. Accumulated experience allows us to contribute to research and development of different kind of products (i.e. biotechnological), what represents a move towards the development of new technologies and the incorporation of appropriate methodologies to current standards. Radiolabeling is still a convenient choice to obtain images of drug distribution and kinetics. With the techniques currently available and those to be used in a near future, we can undertake to the most important researches required.

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El objetivo del presente trabajo fue desarrollar un péptido híbrido, que presente una secuencia capaz de quelatar al 99mTc y otra con afinidad por el receptor de la vitronectina, que permita la detección in vivo de tumores malignos. El marcaje del péptido PICIC3 con 99mTc se realizó de forma directa. Se estudió la estabilidad del complejo en exceso de L-cisteína y en plasma, la unión a proteínas plasmáticas, el coeficiente de partición en el sistema NaCl 0.9 por ciento:n-octanol, la carga del complejo mediante electroforesis y la afinidad por el receptor de la vitronectina se valoró a partir de un ensayo de saturación con membranas de células B16-F10. Se determinó la biodistribución en ratones C57BL/6 con injertos de melanoma B16-F10. Conclusiones: El péptido desarrollado mostró una afinidad satisfactoria por el receptor de la vitronectina y que permitió la detección in vivo de los melanomas múridos del tipo B16-F10 en los ratones injertados.

The aim of the present work was to develop a hybrid peptide, with a sequence for the chelation of 99mTc and other with affinity for the vitronectine receptor, to allow in vivo detection of malignant tumors. 99mTc-labeling of peptide PICIC3 was directly performed. The stability in presence of L-cysteine excess and plasma of the complex, its binding to plasma proteins, the partition coefficient in NaCl 0.9 percent:n-octanol, the charge of the chelate by electrophoresis and the peptide affinity for the vitronectine receptor by a saturation assay using membranes of B16-F10 cells, were studied. Biodistribution in C57BL/6 mice injerted with melanoma B16-F10 was assessed. Conclusions: Developed peptide showed a satisfactory affinity for the vitronectine receptor and allowed in vivo detection of murine melanomas in mice with allografts.

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RESUMEN En este trabajo, se realizó el diseño, optimización, control de calidad de pureza radioquímica, cualidad biológica y estudios de estabilidad en el tiempo de una formulación de macroagregados de albúmina a partir de la desnaturalización controlada de la ASH. Se realizó un diseño experimental con un plan factorial para determinar las condiciones ideales de la formulación. Los resultados mostraron que la concentración de albúmina en el intervalo 515 mg/mL es irrelevante, sin embargo, el pH final de la mezcla y la velocidad de agitación sí fueron relevantes, así como todas las variables de interacción. Con las condiciones seleccionadas en el diseño experimental, se obtuvieron formulaciones con tamaño de partículas con diámetros comprendidos entre 11 y 80 µ, que cumplieron los criterios de pureza radioquímica establecidos internacionalmente, así como la condición de esterilidad y apirogenicidad. La biodistribución en animales de experimentación mostró imágenes pulmonares de buena calidad. Los lotes producidos mantuvieron estabilidad física, radioquímica, biológica y microbiológica durante 90 días de conservación a 4°C.

ABSTRACT The present paper shows the design, optimization, quality control for testing radiochemical purity, biological quality and realtime stability studies of a preparation of macroaggregated albumin from the controlled denaturation of HSA. An experimental design with a factorial plan was carried out in order to determine the ideal conditions for the preparation. The results showed that variations of albumin concentrations within the range of 515 mg/mL were irrelevant, however, the final pH of the mixture and the stirring speed were significant variables, as well as their interaction. Formulations with particles of diameter between 11 and 80 µ were obtained with the conditions selected in the experimental design which met the internationally established criteria for radiochemical purity, sterility and apyrogenicity. The biodistribution in experimental animals showed good quality lung images. The produced lots maintained physical, radiochemical, biological and microbiological stability during 90 days of storage at 4°C.