Person: Bolas Fernández, Francisco
Loading...
First Name
Francisco
Last Name
Bolas Fernández
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Farmacia
Department
Microbiología y Parasitología
Area
Parasitología
Identifiers
13 results
Search Results
Now showing 1 - 10 of 13
Item Transferosomes as nanocarriers for drugs across the skin: Quality by design from lab to industrial scale(International Journal of Pharmaceutics, 2021) Fernández-García, Raquel; Lalatsa, Aikaterini; Statts, Larry; Bolas Fernández, Francisco; Ballesteros Papantonakis, María De La Paloma; Serrano López, Dolores RemediosTransferosomes, also known as transfersomes, are ultradeformable vesicles for transdermal applications consisting of a lipid bilayer with phospholipids and an edge activator and an ethanol/aqueous core. Depending on the lipophilicity of the active substance, it can be encapsulated within the core or amongst the lipid bilayer. Compared to liposomes, transferosomes are able to reach intact deeper regions of the skin after topical administration delivering higher concentrations of active substances making them a successful drug delivery carrier for transdermal applications. Most transferosomes contain phosphatidylcholine (C18) as it is the most abundant lipid component of the cell membrane, and hence, it is highly tolerated for the skin, decreasing the risk of undesirable effects, such as hypersensitive reactions. The most common edge activators are surfactants such as sodium deoxycholate, Tween® 80 and Span® 80. Their chain length is optimal for intercalation within the C18 phospholipid bilayer. A wide variety of drugs has been successfully encapsulated within transferosomes such as phytocompounds like sinomenine or apigenin for rheumatoid arthritis and leukaemia respectively, small hydrophobic drugs but also macromolecules like insulin. The main factors to develop optimal transferosomal formulations (with high drug loading and nanometric size) are the optimal ratio between the main components as well as the critical process parameters for their manufacture. Application of quality by design (QbD), specifically design of experiments (DoE), is crucial to understand the interplay among all these factors not only during the preparation at lab scale but also in the scale-up process. Clinical trials of a licensed topical ketoprofen transferosomal gel have shown promising results in the alleviation of symptons in orthreothritis with non-severe skin and subcutaneous tissue disorders. However, the product was withdrawn from the market which probably was related to the higher cost of the medicine linked to the expensive manufacturing process required in the production of transferosomes compared to other conventional gel formulations. This example brings out the need for a careful formulation design to exploit the best properties of this drug delivery system as well as the development of manufacturing processes easily scalable at industrial level.Item Enhanced bioavailability and anthelmintic efficacy of mebendazole in redispersible microparticles with low-substituted hydroxypropylcellulose(Drug Design, Development and Therapy, 2014) Torre Iglesias, Paloma Marina De La; García Rodríguez, Juan José; Torrado Durán, Guillermo; Torrado Durán, Santiago; Torrado Durán, Susana; Bolas Fernández, FranciscoAntecedentes: el mebendazol (MBZ) es un fármaco extremadamente insoluble y, por lo tanto, poco absorbido y los resultados clínicos variables pueden correlacionarse con las concentraciones sanguíneas. La necesidad de un tratamiento prolongado con dosis altas de este fármaco aumenta el riesgo de efectos adversos. Métodos: En el presente estudio preparamos micropartículas redispersables (RDM) que contienen MBZ, un fármaco oral poco soluble en agua, en diferentes proporciones de hidroxipropilcelulosa poco sustituida (L-HPC). Investigamos las estructuras de micropartículas que emergen espontáneamente tras la dispersión de un RDM en medio acuoso y dilucidamos su influencia en la disolución, y también en su biodisponibilidad oral y eficiencia terapéutica utilizando un modelo murino de infección con el parásito nematodo Trichinella espiralis. Resultados: Se obtuvieron porcentajes elevados de fármaco disuelto con RDM en proporciones 1:2,5 y 1:5 de MBZ:L-HPC. El análisis térmico mostró una amorfización de MBZ en el RDM por la ausencia de un pico de fusión claro de MBZ en las formulaciones. El rápido comportamiento de disolución podría deberse a la disminución de la cristalinidad del fármaco, el rápido tiempo de disolución de portadores como L-HPC, junto con su superior dispersabilidad y excelentes propiedades humectantes. RDM-1:2,5 y RDM-1:5 dieron como resultado un aumento de la concentración plasmática máxima y de los valores de área(s) bajo la curva (AUC)(0-infinito). Asimismo, después de la administración oral de RDM-1:2,5 y RDM-1:5, el AUC(0-infinito) fue 2,67 y 2,97 veces mayor, respectivamente, en comparación con el MBZ puro. La actividad terapéutica, evaluada en el ciclo de vida de Trichinella espiralis, mostró que RDM-1:5 fue el más eficaz para reducir el número de parásitos (4,56 veces) en comparación con el MBZ puro, en estado enquistado. Conclusión: El MBZ: L-HPC RDM podría ser una forma efectiva de mejorar la biodisponibilidad oral y la actividad terapéutica utilizando dosis bajas de MBZ (5 mg/kg), lo que implica un bajo grado de toxicidad para los humanos.Item Self-assembling, supramolecular chemistry and pharmacology of amphotericin B: Poly-aggregates, oligomers and monomers(Journal of Controlled Release, 2021) Raquel Fernández-García; Juan C. Muñoz-García; Matthew Wallace; Laszlo Fabian; González Burgos, Elena María; Gómez-Serranillos Cuadrado, María Pilar; Raposo González, Rafaela; Bolas Fernández, Francisco; Ballesteros Papantonakis, María De La Paloma; Anne Marie Healy; Yaroslav Z. Khimyak; Serrano López, Dolores RemediosAntifungal drugs such as amphotericin B (AmB) interact with lipids and phospholipids located on fungal cell membranes to disrupt them and create pores, leading to cell apoptosis and therefore efficacy. At the same time, the interaction can also take place with cell components from mammalian cells, leading to toxicity. AmB was selected as a model antifungal drug due to the complexity of its supramolecular chemical structure which can self-assemble in three different aggregation states in aqueous media: monomer, oligomer (also known as dimer) and poly-aggregate. The interplay between AmB self-assembly and its efficacy or toxicity against fungal or mammalian cells is not yet fully understood. To the best of our knowledge, this is the first report that investigates the role of excipients in the supramolecular chemistry of AmB and the impact on its biological activity and toxicity. The monomeric state was obtained by complexation with cyclodextrins resulting in the most toxic state, which was attributed to the greater production of highly reactive oxygen species upon disruption of mammalian cell membranes, a less specific mechanism of action compared to the binding to the ergosterol located in fungal cell membranes. The interaction between AmB and sodium deoxycholate resulted in the oligomeric and polyaggregated forms which bound more selectively to the ergosterol of fungal cell membranes. NMR combined with XRD studies elucidated the interaction between drug and excipient to achieve the AmB aggregation states, and ultimately, their diffusivity across membranes. A linear correlation between particle size and the efficacy/toxicity ratio was established allowing to modulate the biological effect of the drug and hence, to improve pharmacological regimens. However, particle size is not the only factor modulating the biological response but also the equilibrium of each state which dictates the fraction of free monomeric form available. Tuning the aggregation state of AmB formulations is a promising strategy to trigger a more selective response against fungal cells and to reduce the toxicity in mammalian cells.Item In vitro effect of new formulations of amphotericin B on amastigote and promastigote forms of Leishmania infantum(International journal of antimicrobial agents, 2007) Ordóñez Gutiérrez, Lara; Espada Fernández, Raquel; Dea Ayuela, María Auxiliadora; Torrado Durán, Juan José; Bolas Fernández, Francisco; Alunda Rodríguez, José MaríaThe in vitro antileishmanial activities of various new amphotericin B (AMB) formulations were investigated, including microspheres of hydrophilic albumin with three AMB aggregation forms (monomeric, dimeric and multiaggregate) and the polymers of polylactic-co-glycolic acid, Resomer RG502 and RG503 with the multiaggregate AMB form. This in vitro study was performed on the extracellular promastigote form and the intracellular amastigote form of a canine strain of Leishmania infantum (UCM 20) using the infected J774 murine macrophage-like cell line. Albumin-encapsulated forms did not show any toxicity for murine cells and had lower median effective concentration (EC50) values (ca. 0.003 microg/mL) for L. infantum amastigotes than free formulations (0.03 microg/mL). In addition, the aggregation state of AMB had a notable effect on the antileishmanial activity of the drug. Results obtained in vitro point towards interest in monomeric AMB encapsulated in microspheres in the chemotherapeutic control of leishmaniasis.Item Can Amphotericin B and Itraconazole be co-delivered orally? Tailoring Oral Fixed-Dose Combination Coated Granules for Systemic Mycoses(European Journal of Pharmaceutics and Biopharmaceutics, 2023) Fernández García, Raquel; Walsh, David; O'Connell, Peter; Slowing Barillas, Karla Verónica; Raposo González, Rafaela; Ballesteros Papantonakis, María De La Paloma; JImenez-Cebrian, Aurora; Chamorro Sancho, Manuel J.; Bolas Fernández, Francisco; Healy, Anne Marie; Serrano López, Dolores RemediosThe incidence and prevalence of invasive fungal infections have increased significantly over the last few years, leading to a global health problem due to the lack of effective treatments. Amphotericin B (AmB) and itraconazole (ITR) are two antifungal drugs with different mechanisms of action. In this work, AmB and ITR have been formulated within granules to elicit an enhanced pharmacological effect, while enhancing the oral bioavailability of AmB. A Quality by Design (QbD) approach was utilised to prepare fixed-dose combination (FDC) granules consisting of a core containing AmB with functional excipients, such as inulin, microcrystalline cellulose (MCC), chitosan, sodium deoxycholate (NaDC) and Soluplus® and polyvinyl pyrrolidone (PVP), coated with a polymeric layer containing ITR with Soluplus® or a combination of Poloxamer 188 and hydroxypropyl methyl cellulose-acetyl succinate (HPMCAS). A Taguchi designs of experiments (DoE) with 7 factors and 2 levels was carried out to understand the key factors impacting on the physicochemical properties of the formulation followed by a Box-Behnken design with 3 factors in 3 levels chosen to optimise the formulation parameters. The core of the FDC granules was obtained by wet granulation and later coated using a fluidized bed. In vitro antifungal efficacy was demonstrated by measuring the inhibition halo against different species of Candida spp., including C. albicans (24.19-30.48 mm), C. parapsilosis (26.38-27.84 mm) and C. krusei (11.48-17.92 mm. AmB release was prolonged from 3 to 24 hours when the AmB granules were coated. In vivo in CD-1 male mice studies showed that these granules were more selective towards liver, spleen and lung compared to kidney (up to 5-fold more selective in liver, with an accumulation of 8.07 µg AmB/g liver after twice-daily 5 days administration of F2), resulting in an excellent oral administration option in the treatment of invasive mycosis. Nevertheless, some biochemical alterations were found, including a decrease in blood urea nitrogen (~17 g/dl) and alanine aminotransferase (<30 U/l) and an increase in the levels of bilirubin (~0.2 mg/dl) and alkaline phosphatase (<80 U/l), which could be indicative of a liver failure. Once-daily regimen for 10 days can be a promising therapy.Item New amphotericin B-gamma cyclodextrin formulation for topical use with synergistic activity against diverse fungal species and Leishmania spp.(International journal of pharmaceutics, 2014) Ruiz, Helga K.; Serrano López, Dolores Remedios; Dea Ayuela, María Auxiliadora; Bilbao Ramos, Pablo Estanislao; Bolas Fernández, Francisco; Torrado Durán, Juan José; Molero, GloriaAmphotericin B (AmB) has a broad antifungal and leishmanicidal activity with low incidence of clinical resistance. Its parenteral administration has high risk of nephrotoxicity that limits its use. In order to treat cutaneous infections, AmB topical administration is a safer therapy because of the low systemic absorption of the drug across mucous membranes. Moreover, in some developing countries both fungal topical infections and cutaneous leishmaniasis are an important health problem. The aim of this work is to formulate a topical amphotericin preparation and test its in vitro antifungal (against 11 different fungal species) and antileishmanial activity. γ-Cyclodextrin (γ-CD) was chosen to solubilise AmB. Furthermore, γ-CD has shown a synergistic effect on membrane destabilization with AmB. Topical novel formulations based on AmB-CD complex have exhibited greater antifungal activity (48%, 28% and 60% higher) when compared to AmB Neo-Sensitabs(®) disks, AmB dissolved in dimethyl sulfoxide (DMSO) and Clotrimazole(®) cream, respectively. Furthermore, AmB-CD methyl cellulose gel has shown significantly higher inhibition activity on biofilm formation, larger penetration through yeast biofilms and higher fungicidal activity on biofilm cells compared to AmB dissolved in DMSO. In addition, AmB-CD gel exhibited both high in vitro leishmanicidal efficacy with wider therapeutic index (between 2 and 8-fold higher than AmB deoxycholate depending on Leishmania spp.) and also in vivo activity in an experimental model of cutaneous leishmaniasis. These results illustrate the feasibility of a topical AmB formulation easy to prepare, physicochemically stable over 6 months, safe and effective against diverse fungal and parasitic cutaneous infections.Item Evaluating the Potential of Ursolic Acid as Bioproduct for Cutaneous and Visceral Leishmaniasis(Molecules, 2020) Bilbao Ramos, Pablo Estanislao; Serrano López, Dolores Remedios; Ruiz Saldaña, Helga Karina; Torrado Durán, Juan José; Bolas Fernández, Francisco; Dea Ayuela, María AuxiliadoraLeishmaniasis affects around 12 million people worldwide and is estimated to cause the ninth-largest disease burden. There are three main forms of the disease, visceral (VL), cutaneous (CL), and mucocutaneous (MCL), leading to more than one million new cases every year and several thousand deaths. Current treatments based on chemically synthesized molecules are far from ideal. In this study, we have tested the in vitro and in vivo efficacy of ursolic acid (UA), a multifunctional triterpenoid with well-known antitumoral, antioxidant, and antimicrobial effects on different Leishmania strains. The in vitro antileishmanial activity against the intracellular forms was six and three-fold higher compared to extracellular forms of L. amazonensis and L. infantum, respectively. UA also showed to be a potent antileishmanial drug against both VL and CL manifestations of the disease in experimental models. UA parenterally administered at 5 mg/kg for seven days significantly reduced the parasite burden in liver and spleen not only in murine acute infection but also in a chronic-infection model against L. infantum. In addition, UA ointment (0.2%) topically administered for four weeks diminished (50%) lesion size progression in a chronic infection model of CL caused by L. amazonensis, which was much greater than the effect of UA formulated as an O/W emulsion. UA played a key role in the immunological response modulating the Th1 response. The exposure of Leishmania-infected macrophages to UA led to a significant different production in the cytokine levels depending on the Leishmania strain causing the infection. In conclusion, UA can be a promising therapy against both CL and VL.Item Repurposing Butenafine as An Oral Nanomedicine for Visceral Leishmaniasis(Pharmaceutics, 2019) Bezerra Souza, Adriana; Fernández García, Raquel; Rodrigues, Gabriela F.; Bolas Fernández, Francisco; Dalastra Laurenti, Marcia; Passero, Luiz Felipe; Lalatsa, Aikaterini; Serrano López, Dolores RemediosLeishmaniasis is a neglected tropical disease a_ecting more than 12 million people worldwide, which in its visceral clinical form (VL) is characterised by the accumulation of parasites in the liver and spleen, and can lead to death if not treated. Available treatments are not well tolerated due to severe adverse e_ects, need for parenteral administration and patient hospitalisation, and long duration of expensive treatments. These treatment realities justify the search for new e_ective drugs, repurposing existing licensed drugs towards safer and non-invasive cost-e_ective medicines for VL. In this work, we provide proof of concept studies of butenafine and butenafine self-nanoemulsifying drug delivery systems (B-SNEDDS) against Leishmania infantum. Liquid B-SNEDDS were optimised using design of experiments, and then were spray-dried onto porous colloidal silica carriers to produce solid-B-SNEDDS with enhanced flow properties and drug stability. Optimal liquid B-SNEDDS consisted of Butenafine:Capryol 90:Peceol:Labrasol (3:49.5:24.2:23.3 w/w), which were then sprayed-dried with Aerosil 200 with a final 1:2 (Aerosil:liquid B-SNEDDS w/w) ratio. Spray-dried particles exhibited near-maximal drug loading, while maintaining excellent powder flow properties (angle of repose <10_) and sustained release in acidic gastrointestinal media. Solid-B-SNEDDS demonstrated greater selectivity index against promastigotes and L. infantum-infected amastigotes than butenafine alone. Developed oral solid nanomedicines enable the non-invasive and safe administration of butenafine as a cost-e_ective and readily scalable repurposed medicine for VL.Item Ultradeformable Lipid Vesicles Localize Amphotericin B in the Dermis for the Treatment of Infectious Skin Diseases(ACS Infectious diseases, 2020) Fernández García, Raquel; Statts, Larry; Jesus, Jéssica A. De; Dea-Ayuela, María Auxiliadora; Bautista Chávez, Liliana; Simão, Rubén; Bolas Fernández, Francisco; Ballesteros Papantonakis, María De La Paloma; Laurenti, Marcia Dalastra; Passero, Luiz F. D.; Lalatsa, Aikaterini; Serrano López, Dolores RemediosCutaneous fungal and parasitic diseases remain challenging to treat, as available therapies are unable to permeate the skin barrier. Thus, treatment options rely on systemic therapy,which fail to produce high local drug concentrations but can lead tosignificant systemic toxicity. Amphotericin B (AmB) is highly efficacious in the treatment of both fungal and parasitic diseases such as cutaneous leishmaniasis but is reserved for parenteraladministration in patients with severe pathophysiology. Here, we have designed and optimized AmB-transfersomes [93.5% encapsulationefficiency, 150 nm size, and good colloidal stability (−35.02mV)] that can remain physicochemically stable (>90% drug content)at room temperature and 4 °C over 6 months when lyophilized and stored under desiccated conditions. AmB-transfersomes possessed good permeability across mouse skin (4.91 ± 0.41 μg/cm2/h) and 10-fold higher permeability across synthetic Strat-M membranes. In vivo studies after a single topical application in mice showed permeability and accumulation within the dermis (>25 μg AmB/g skin 6 h postadministration), indicating the delivery of therapeutic amounts of AmB for mycoses and cutaneous leishmaniasis, while a single daily administration in Leishmania (Leishmania) amazonensis infected mice over 10 days, resulted in excellent efficacy (98% reduction in Leishmania parasites). Combining the application of AmB-transfersomes with metallic microneedles in vivo increased the levels in the SC and dermis but was unlikely to elicit transdermal levels. In conclusion, AmB-transfersomes are promising and stable topical nanomedicines that can be readily translated for parasitic and fungal infectious diseases.Item Changes in the proteome and infectivity of Leishmania infantum induced by in vitro exposure to a nitric oxide donor(International journal of medical microbiology, 2009) Dea Ayuela, María Auxiliadora; Ordóñez Gutiérrez, Lara; Bolas Fernández, FranciscoLeishmania species are protozoan parasites that exhibit an intracellular amastigote form within mammalian macrophages and an extracellular promastigote form inside the sandfly vector. The generation of nitric oxide (NO) upon activation of macrophages is surely the principal killing effector of intracellular amastigotes but little is known about the potential action of NO against the promastigote phase during its multiplication inside the digestive tract of the sandfly vector. Therefore, we have approached this issue by using an in vitro model to study the effect of an NO donor, 3-morpholinosydnonimine (SIN-1), on the proteome and infectivity of promastigotes of Leishmania infantum. Exposure of promastigotes to SIN-1 during its logarithmic growth phase caused a dramatic effect on parasite protein expression and viability, consequently killing about 60–70% of the promastigotes. The significant changes in the proteome included the over-expression of enolase, peroxidoxin precursors, and heat-shock protein 70 (HSP70), underexpression of 20S proteasome alpha 5 unit, and phosphomannomutase and induced expression of 3-hydroxy-3-methyglutaryl-CoA (HMG-CoA) synthase and prostaglandine f2-alpha (PGD2) synthase. Interestingly, promastigotes that resisted treatment showed enhanced infectivity to J774 macrophages in comparison to the controls. This finding together with the appearance of the PGD2S and an over-expression of HSP70 isoforms in treated promastigotes led us to speculate the existence of NO-mediated programmed cell death (PCD) events as a potential mechanism of population regulation and selection of properly infecting forms that predominantly operate on the promastigote stage.