Desenvolupament del nou mètode DELOS (R) de cristal·lització amb fluids comprimits : estudi a nivell molecular dels seus fonaments /

Consultable des del TDXA portada: Departament de Nanociència Molecular i Materials Orgànics. Institut de Ciència de Materials de Barcelona (CSIC)Títol obtingut de la portada digitalitzadaLa present Tesi Doctoral es troba emmarcada dins del camp de les tecnologies de processament de materials emprant fluids comprimits (FCs), i en concret dins dels mètodes de preparació de sòlids finament dividits que es basen en la utilització d'aquest tipus de fluids. Els mètodes de cristal·lització o preparació de sòlids micro o nanoparticulats que utilitzen FCs es caracteritzen perquè la modulació de la variable pressió fa possible que es generin canvis abruptes en la capacitat solvatadora del medi dissolvent que es transmeten de manera molt homogènia en tot el si de la dissolució. Com a conseqüència s'obtenen perfils de sobresaturació amb elevats valors de velocitat de generació de sobresaturació i valors també elevats de sobresaturació màxima, i que per tant condueixen a un procés amb predomini del fenomen de nucleació per sobre del fenomen de creixement cristal·lí. Així, en el present treball es mostra el procés de desenvolupament del nou mètode DELOS® de preparació de materials finament dividits que es basa en la utilització del FC CO2 com a cosolvent en dissolucions orgàniques expandides amb CO2. En una primera part de la tesi, es demostra que el fenomen de cristal·lització associat al nou mètode DELOS® és conseqüència del refredament que sofreix una dissolució expandida amb CO2 del compost a cristal·litzar, quan es realitza la seva despressurització fins a pressió atmosfèrica. Tal i com es prova, el refredament observat és conseqüència de l'evaporació del CO2 de la dissolució expandida i el procés associat d'intercanvi de calor que es produeix, i que per tant, només depèn, per a un determinat sistema, del contingut en CO2 d'aquesta dissolució. En aquesta primera part de la tesi també es demostra que és viable aplicar el mètode DELOS a materials de naturalesa variada. Els experiments realitzats en aquest sentit permeten veure que s'obté un control molt significatiu de les característiques del sòlid particulat produït, tant en termes de mida de partícula com en termes de cristal·linitat i naturalesa polimòrfica. En una segona part de la tesi es mostra com la utilització de tensoactius en el nou mètode desenvolupat és beneficiós per a la reducció dels fenòmens d'agregació i aglomeració de partícules i per tant per a l'estabilització d'aquestes partícules com a entitats discretes. També es mostra que és possible la directa estabilització de partícules obtingudes per despressurització de dissolucions expandides directament sobre dissolucions aquoses amb un cert contingut en tensoactiu, la qual cosa pot permetre la preparació de nanosuspensions susceptibles per a ser utilitzades en l'administració de principis actius amb baixa solubilitat en aigua. Aquesta possibilitat ha permès desenvolupar el nou mètode anomenat DELOS-SUSP. L'última part de la tesi presenta estudis realitzats a escala molecular en relació a dissolucions expandides amb CO2, els quals permeten obtenir informació molt rellevant en relació als sistemes utilitzats en l'aplicació del mètode DELOS®. En aquests estudis s'utilitza la tècnica d'espectroscòpia d'Infraroig a alta pressió en combinació amb l'aproximació semiempírica Linear Solvation Energy Relationships (LSER) i càlculs teòrics ab-initio. Els resultats obtinguts permeten determinar l'evolució de l'esfera de solvatació de soluts dissolts en dissolvents expandits amb CO2, la naturalesa de les interaccions solut-dissolvent que s'hi donen i l'efecte, a nivell molecular, que té la presència de CO2 en els sistemes subjectes a estudi.The present work is devoted to the field of materials processing using compressed fluids (CFs), and more precisely, it deals with methods for the preparation of finely divided solid materials in CFs. It is well known that the methods for the preparation of micro or nano-particulate solids using CFs take advantage of pressure changes to produce abrupt changes on the solvating power of solvent media which are transmitted very homogeneously along all the points of the solution. This makes possible to obtain a supersaturation profile characterized by high rates of supersaturation generation and also high values of maximum supersaturations, which define a process with high promotion of nucleation phenomena over crystal growth. In the present work, it is shown the process of development of the new DELOS method for the production of finely divided materials based on the use of the CF CO2 as cosolvent in CO2-expanded organic solutions. In a first part of the thesis, it is demonstrated that the crystallisation phenomenon associated with the DELOS process is consequence of the cooling effect experienced by a CO2-expanded solution when it is depressurized to atmospheric pressure. It is proved that this cooling effect is consequence of the evaporation of CO2 from the expanded solution and the derived heat exchange process. Therefore, the temperature decrease observed is only function, for a certain system, of the CO2 content of this solution. In this first part of the thesis it is also demonstrated the viability to apply the DELOS process to materials of different nature. The experiments realized on this issue allow to see that it is possible to tune and control solid characteristics such as particle size, cristallinity and polymorphic nature. In a second part of the thesis, it is shown that the use of surfactants in the new process developed can contribute to reduce the occurrence of aggregation and agglomeration phenomena and hence promote the stability of the particles obtained. It is also shown that the stabilization of particles in water as nanosuspensions can be achieved by performing the depressurization of a CO2-expanded solution directly into a solution of a surfactant in water. This possibility has lead to the development of a new process for the preparation of nanosuspensions that has been called DELOS-SUSP. Finally, the last part of the thesis presents molecular level studies concerning CO2-expanded solutions which enable to obtain very relevant information related to the systems employed for the application of the DELOS process. In these studies, High pressure IR spectroscopy is employed in combination with the solvent effects treatment Linear Solvation Energy Relationships and theoretical abinitio calculations. The results obtained permit to determine the evolution of the solvation shell of solutes dissolved in CO2-expanded solutions, the nature of solute-solvent interactions that take place in this region and the molecular level effect that causes CO2 in the systems subject of study

Procedimiento para la obtención de micro- o nanopartículas sólidas

Procedimiento para la obtencion de micro- o nanoparticulas solidas. La invencion proporciona un nuevo procedimiento para la obtencion de micro- o nanoparticulas solidas con estructura homogenea. Se proporciona un procedimiento que permite obtener micro- o nanoparticulas solidas de estructura homogenea, con un tamano de particula inferior a 10 ƒÊm donde el compuesto solido procesado revela la naturaleza, cristalina, amorfa, polimorfica, etc..., propia del compuesto de partida. De acuerdo con la invencion se proporciona un procedimiento que ademas permite obtener micro- o nanoparticulas solidas con una morfologia sustancialmente esferoidal.Peer reviewedConsejo Superior de Investigaciones Científicas (Esspaña), Centro de Investigación Biomédica en Red en Bioingeniería Biomateriales y Nanomedicina (CIBER-BBN)A1 Solicitud de patente con informe sobre el estado de la técnic

Lipid-based nanovesicles for nanomedicine

Molecular self-assembly has enabled the fabrication of biologically inspired, advanced nanostructures as lipid-based nanovesicles (L-NVs). The oldest L-NVs, liposomes, have been widely proposed as potential candidates for drug delivery, diagnostic and/or theranostic applications and some liposome-based drug products have already stepped from the lab-bench to the market. This success is attributed to their ability to encapsulate both hydrophobic and/or hydrophilic molecules, efficiently carry and protect them within the body and finally deliver them at the target site. These positive features are also coupled with high biocompatibility. However, liposomes still present some unsolved drawbacks, such as poor colloidal stability, short shelf-life, restricted and expensive conditions of preparation because of the inherent nature of their fundamental constituents (phospholipids). The new tools available in the self-assembly of controlled molecules have significantly advanced the field of L-NV design and synthesis, and nonliposomal L-NVs have been recently developed; this new generation of nanovesicles can represent a paradigm shift in nanomedicine: they may complement liposomes, showing their advantages and overcoming most of their drawbacks. Clearly, being still young, their rocky way to the clinic first and then to the market has just started and it is still long, but they have all the potentialities to reach their objective target. The purpose of this review is to first present the large plethora of L-NVs available, focusing on this new generation of non-liposomal L-NVs and showing their similarities and differences with respect to their ancestors (liposomes). Since the overspread of a nanomaterial to the market is also strongly dependent on the availability of technological-scale preparation methods, we will also extensively review the current approaches exploited for L-NV production. The most cutting-edge approaches based on compressed fluid (CF) technologies will be highlighted here since they show the potential to represent a game-change in the production of L-NVs, favouring their step from the bench to the market. Finally, we will briefly discuss L-NV applications in nanomedicine, looking also for their future perspectives.This work was financially supported by MINECO (DGI), Spain, grants BE-WELL CTQ2013-40480-R, TERARMET (RTC-2014-2207-1), QUATFORFRAG (RTC-2014-2183-5) and UNDERLIPIDS (RTC-2015-3303-1), and by AGAUR, Generalitat de Catalunya, “Grant 2014-SGR-17“. N.G. acknowledges the European Commission (EC) (FP7-PEOPLE-2013-Initial Training Networks (ITN) ‘‘NANO2FUN’’ Project No. 607721) for their Postdoctoral contract. The authors appreciate support from LIPOCELL project financed by CIBER-BBN and Praxis Pharmaceuticals and also acknowledge the financial support from Instituto de Salud Carlos III, through “Acciones CIBER”. The Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN) is an initiative funded by the VI National R&D&I Plan 2008–2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions and financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund.Peer reviewe

α-Galactosidase-A Loaded-Nanoliposomes with Enhanced Enzymatic Activity and Intracellular Penetration

Lysosomal storage disorders (LSD) are caused by lysosomal dysfunction usually as a consequence of deficiency of a single enzyme required for the metabolism of macromolecules, such as lipids, glycoproteins, and mucopolysaccharides. For instance, the lack of α-galactosidase A (GLA) activity in Fabry disease patients causes the accumulation of glycosphingolipids in the vasculature leading to multiple organ pathology. Enzyme replacement therapy, which is the most common treatment of LSD, exhibits several drawbacks mainly related to the instability and low efficacy of the exogenously administered therapeutic enzyme. In this work, the unprecedented increased enzymatic activity and intracellular penetration achieved by the association of a human recombinant GLA to nanoliposomes functionalized with Arginine-Glycine-Aspartic acid (RGD) peptides is reported. Moreover, these new GLA loaded nanoliposomes lead to a higher efficacy in the reduction of the GLA substrate named globotriasylceramide in a cellular model of Fabry disease, than that achieved by the same concentration of the free enzyme. The preparation of these new liposomal formulations by DELOS-SUSP, based on the depressurization of a CO-expanded liquid organic solution, shows the great potential of this CO-based methodology for the one-step production of protein-nanoliposome conjugates as bioactive nanomaterials with therapeutic interest. α-galactosidase-A-(GLA)-loaded nano-liposomes functionalized with Arginine-Glycine-Aspartic acid (RGD) peptides are successfully prepared by using compressed CO. This nanoformulation shows an unprecedented increase of the GLA enzymatic activity and intracellular penetration, in comparison to the free enzyme. Moreover, these nanoconjugates lead to a higher efficacy in the reduction of the GLA substrate named globotriasylceramide (Gb3), in a cellular model of Fabry disease, than that achieved by the free enzyme.We acknowledge financial support from Instituto de Salud Carlos III, through “Acciones CIBER”. The Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN) is an initiative funded by the VI National R&D&I Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions and financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund. The authors appreciate the financial support through the “Development of nanomedicines for enzymatic replacement therapy in Fabry disease” project, granted by the Fundació Marató TV3, BE-WELL (CTQ2013-40480-R) project granted by DGI (Spain), GenCat (2014-SGR-17) project financed by DGR (Catalunya), LIPOCELL project financed by CIBER-BBN and Praxis Pharmaceuticals, TERARMET (RTC-2014-2207-1) project financed by MEC (Spain) and excellence Grant CTS-6270 financed by “Junta de Andalucía>. The authors wish also to thank the Microscopy Service of UAB, especially Pablo Castro for the technical support in taking the Cryo-TEM images, and Mª Eugenia López Sánchez and Natalia García Aranda for their technical assistance in the in vitro activity assays. AV is recipient of an ICREA Academia (Generalitat de Catalunya) award. We acknowledge EMBL and Electra synchrotrons for beamtime allocation, and H. Amenitsch, B. Marmilori and B. Sartori for technical support at the SAXS beamline. Access to the synchrotron facility is supported by the BioStructX program (nº BIOSTRUCTX_1093). We also acknowledge computer time at the Minotauro-BSC supercomputer from the Red Española de Supercomputación (RES).Peer Reviewe

DELOS Nanovesicles-Based Hydrogels: An Advanced Formulation for Topical Use

Topical delivery has received great attention due to its localized drug delivery, its patient compliance, and its low risk for side effects. Recent developments have focused on studying new drug delivery systems as a strategy for addressing the challenges of current topical treatments. Here we describe the advances on an innovative drug delivery platform called DELOS nanovesicles for topical drug delivery. Previously, the production of DELOS nanovesicles demonstrated potentiality for the topical treatment of complex wounds, achieving well-tolerated liquid dispersions by this route. Here, research efforts have been focused on designing these nanocarriers with the best skin tolerability to be applied even to damaged skin, and on exploring the feasibility of adapting the colloidal dispersions to a more suitable dosage form for topical application. Accordingly, these drug delivery systems have been efficiently evolved to a hydrogel using MethocelTM K4M, presenting proper stability and rheological properties. Further, the integrity of these nanocarriers when being gellified has been confirmed by cryo-transmission electron microscopy and by Förster resonance energy transfer analysis with fluorescent-labeled DELOS nanovesicles, which is a crucial characterization not widely reported in the literature. Additionally, in vitro experiments have shown that recombinant human Epidermal Growth Factor (rhEGF) protein integrated into gellified DELOS nanovesicles exhibits an enhanced bioactivity compared to the liquid form. Therefore, these studies suggest that such a drug delivery system is maintained unaltered when hydrogellified, becoming the DELOS nanovesicles-based hydrogels, an advanced formulation for topical use.This research was funded by Industrial Doctorates Plan of Agaur-Generalitat de Catalunya (2018 DI 057). This work was also co-financed by European Regional Development Funds from the European Union within the framework of the operative FEDER program in Catalonia 2014–2020, by the specific project NANONAFRES (COMRDI15-1-0023). We thank the Spanish Ministry of Science and Innovation (MICINN) for financing MOL4BIO project through grant No. PID2019-105622RB-I00. This project has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No 953110. ICMAB-CSIC members acknowledge support from the MINECO through the Severo Ochoa Program for Centers of Excellence in R&D (SEV-2015-0496 and CEX2019-000917-S), and through the State Program for R&D&i (2016), orientated to the Society Challenges, modality RETOS-Collaboration 2016, and co-financed by FEDER funds from the European Commission (grant No. RTC-2016-4567-1, Nano4Derm). ICMAB-CSIC members also thank Agaur agency of Generalitat de Catalunya for their support through grant No. 2017-SGR-918. DELOS nanovesicle production and their physicochemical characterization was performed by the ICTS “NANBIOSIS”, more specifically in the Biomaterial Processing and Nanostructuring Unit (U6), Unit of the CIBER in Bioengineering, Biomaterials and Nanomedicne (CIBER-BBN) located at the Institute of Materials Science of Barcelona (ICMAB-CSIC). This work has been done in the framework of the L.B.-H. doctorate in Materials Science of the Universitat Autonoma de Barcelona.With funding from the Spanish government through the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000917-S).Peer reviewe

Procedimiento para la precipitación de partículas sólidas finamente divididas

Referencia OEPM: P200002129.-- Fecha de solicitud: 25/08/2000.-- Titular: Sociedad Española de Carbutos Metálicos, S.A.El procedimiento para la precipitación de partículas sólidas finamente divididas comprende: a) Disolver un compuesto C en un fluido A para obtener una solución A; b) Termostatizar dicha solución A a una temperatura comprendida entre -50°C y 200°C; c) Añadir a dicha solución A un fluido B hasta alcanzar una presión P, y se caracteriza por el hecho de que dicho fluido B a una presión P es miscible con dicha solución A y actúa como cosolvente para formar una solución AB; y d) Reducir la presión de dicha solución AB para que tenga lugar la precipitación de dicho compuesto C. El procedimiento permite obtener partículas de tamaño medio inferior a 20 μm, generalmente inferior a 10 μm, con una distribución estrecha de tamaño comprendida entre 1 y 100 μm, generalmente entre 1 y 20 μm, a partir de una solución, y no una mezcla, que contiene el compuesto a precipitar.Peer reviewe

Procedimiento para la obtención de micro- o nanopartículas sólidas

Procedimiento para la obtencion de micro- o nanoparticulas solidas. La invencion proporciona un nuevo procedimiento para la obtencion de micro- o nanoparticulas solidas con estructura homogenea. Se proporciona un procedimiento que permite obtener micro- o nanoparticulas solidas de estructura homogenea, con un tamano de particula inferior a 10 ƒÊm donde el compuesto solido procesado revela la naturaleza, cristalina, amorfa, polimorfica, etc..., propia del compuesto de partida. De acuerdo con la invencion se proporciona un procedimiento que ademas permite obtener micro- o nanoparticulas solidas con una morfologia sustancialmente esferoidal.Peer reviewedConsejo Superior de Investigaciones Científicas (Esspaña), Centro de Investigación Biomédica en Red en Bioingeniería Biomateriales y Nanomedicina (CIBER-BBN)A1 Solicitud de patente con informe sobre el estado de la técnic

Vesicles based on glucose-derived surfactants and phytosterols

It is provided a vesicle comprising at least one phytosterol and at least one glucose-derived surfactant. Further, a composition comprising a plurality of vesicles as defined herein is also provided. The vesicle and the composition comprising a plurality of vesicles are used in the manufacture of a suspension, as a delivery system or as medicament. Further, a method for the manufacture of a vesicle as described herein is also providedPeer reviewedNanomol TechnologiesA1 Solicitud de patente con informe sobre el estado de la técnic

Functionalized liposomes useful for the delivery of bioactive compounds

Ventosa Rull, Leonor et al.[EN] The invention relates to conjugates in which a sterol is functionalized by an ether bond with a water-soluble polymer to which a guiding ligand is bound. These conjugates improve the physico-chemical and delivery properties of their carrying vesicles, making these more stable, homogeneous and effective. A method for their preparation, a pharmaceutical composition containing said liposomes, and their therapeutic use are described as wellPeer reviewedConsejo Superior de Investigaciones Científicas, Fundació Parc Científic de Barcelona, Centro de Investigaciones Biomédicas en Red, Fundació Institut de Recerca Biomédica, Universitat de Barcelona, Fundació Hospital Universitari Vall d'Hebron, Universitat Autónoma de BarcelonaA1 Solicitud de patente con informe sobre el estado de la técnic

Composición de gentamicina, un surfactante aniónico, y un copolímero

Composición de gentamicina, un surfactante aniónico, y un copolímero. La invención se refiere a una composición que comprende gentamicina, un surfactante aniónico, y un copolímero de metil vinil éter y un derivado de ácido maleico, en donde la gentamicina y el surfactante aniónico forman un complejo iónico. Dicha composición se prepara mediante a) la disolución de un complejo iónico de gentamicina y un surfactante aniónico, y un copolímero de metil vinil éter y un derivado de ácido maleico, en un disolvente orgánico, en cuyo disolvente orgánico el complejo iónico y el copolímero son solubles; b) el pulverizado de la disolución resultante de la etapa a) en un fluido comprimido, que actúa como antisolvente y causa la precipitación de un sólido; c) la separación del sólido resultante de la etapa b). La composición obtenida tiene utilidad como medicamento, particularmente para el tratamiento de una enfermedad causada por una bacteria seleccionada del grupo de Salmonella spp., Legionella pneumophila, Staphylococcus aureus, Mycobacterium spp., Brucella spp., Francisella tularensis, Bartonella spp, y Listeria monocytogenes.Peer reviewedConsejo Superior de Investigaciones Científicas (España), Instituto Científico y tecnológico de Navarra SAB1 Patente sin examen previ