Latanoprost-loaded nanotransfersomes designed for scalp administration enhance keratinocytes proliferation.

Latanoprost (LAT) has been shown to have a hypertrichotic effect, which makes it a promising candidate for alopecia treatments. For the first time, LAT has been encapsulated in nanotransfersomes in order to increase its efficacy. Ex vivo skin biodistribution was studied by confocal laser microscopy both in human scalp and pig skin. Results showed that nanotransfersomes increase the penetration of two different fluorochromes, with similar patterns in both species, compared with fluorochrome solutions containing no nanotransfersomes. Nanotransfersomes were stable under accelerated conditions (40 °C/75% RH) and long-term conditions (25 °C/60% RH) for up to 1 year, with no differences in vesicle size and polydispersity when LAT was loaded. Nanotransfersomes increased the LAT cell proliferation effect in HaCaT cell via MAPK signaling pathway. Collectively, our results demonstrate LAT-nanotransfersomes formulation could be a promising therapy for hair growth disorders

Dexamethasone-Loaded Lipomers: Development, Characterization, and Skin Biodistribution Studies

Follicular targeting has gained more attention in recent decades, due to the possibility of obtaining a depot effect in topical administration and its potential as a tool to treat hair follicle-related diseases. Lipid core ethyl cellulose lipomers were developed and optimized, following which characterization of their physicochemical properties was carried out. Dexamethasone was encapsulated in the lipomers (size, 115 nm; polydispersity, 0.24; zeta-potential (Z-potential), +30 mV) and their in vitro release profiles against dexamethasone in solution were investigated by vertical diffusion Franz cells. The skin biodistribution of the fluorescent-loaded lipomers was observed using confocal microscopy, demonstrating the accumulation of both lipomers and fluorochromes in the hair follicles of pig skin. To confirm this fact, immunofluorescence of the dexamethasone-loaded lipomers was carried out in pig hair follicles. The anti-inflammatory (via TNFα) efficacy of the dexamethasone-loaded lipomers was demonstrated in vitro in an HEK001 human keratinocytes cell culture and the in vitro cytotoxicity of the nanoformulation was investigated

Repurposing Disulfiram as an Antimicrobial Agent in Topical Infections

Antimicrobial drugs applied topically offer several advantages. However, the widespread use of antibiotics has led to increasing antimicrobial resistance. One interesting approach in the drug discovery process is drug repurposing. Disulfiram, which was originally approved as an anti-alcoholism drug, offers an attractive alternative to treat topical multidrug resistance bacteria in skin human infections. This study aimed to evaluate the biopharmaceutical characteristics of the drug and the effects arising from its topical application in detail. Microdilution susceptibility testing showed antibacterial activity against Gram-positive bacteria Staphylococcus aureus and Streptococcus pyogenes. Dermal absorption revealed no permeation in pig skin. The quantification of the drug retained in pig skin demonstrated concentrations in the stratum corneum and epidermis, enough to treat skin infections. Moreover, in vitro cytotoxicity and micro-array analyses were performed to better understand the mechanism of action and revealed the importance of the drug as a metal ion chelator. Together, our findings suggest that disulfiram has the potential to be repurposed as an effective antibiotic to treat superficial human skin infections

La psoriasis

Treballs d'Educació Farmacèutica als ciutadans. Unitat Docent d'Estades en Pràctiques Tutelades. Facultat de Farmàcia, Universitat de Barcelona. Curs: 2016-2017. Tutors: David Berlana, Ignasi Cardona Pascual i Marian March Pujol.La psoriasi és una malaltia dermatològica crònica no contagiosa que es caracteritza principalment per lesions a la pell, tot i que pot afectar qualsevol part del cos, inclosos el cuir cabellut, les mans, els peus, els genitals i les ungles. A més, afecta a la salut emocional. És una malaltia autoimmunitària, de manera que una part del sistema immunitari està sobreactivada, fet que provoca que hi hagi una inflamació persistent i una acceleració del procés de renovació de la pell. Les cèl·lules de la pell creixen massa ràpid, i això fa que s’acumulin a la superfície a un ritme molt superior al de la descamació de la pell, donant lloc a una inflamació i envermelliment al voltant de la zona afectada i a l’aparició d’unes plaques de descamació. La seva etiologia és multifactorial, donat que intervenen: - Factors ambientals: estrès, fred, obesitat, traumatismes, infeccions, alcohol, tabac i drogues. - Factors genètics: tot i que no es coneix com s’hereta la malaltia, es creu que la genètica juga un paper clau en el seu desenvolupament. - Factors immunològics És molt important tenir uns hàbits de vida saludables: cuidar-se, fer exercici de forma regular, menjar bé i controlar la pressió arterial

The Efficacy and Biopharmaceutical Properties of a Fixed-Dose Combination of Disulfiram and Benzyl Benzoate

Scabies and hair lice are parasitic diseases that affect human skin and hair, respectively. The incidence and resistances of these infections are increasing. Tenutex® (disulfiram and benzyl benzoate emulsion) is an alternative to standard insecticides to avoid resistances. The aim of the work is to evaluate the transdermal absorption and the in vitro efficacy against scabies and hair lice after different exposition times. Dermatomed human skin was used to assess the dermal absorption using a validated High Performance Liquid Chromatography (HPLC) method. HEK001 keratinocytes were used to evaluate the cytotoxicity of benzyl benzoate. Only benzyl benzoate was able to cross the skin, but it did not show cytotoxicity at any of the tested concentrations. The product efficacy was tested on Psoroptes ovis after direct contact and after administration on sheep skin explants at different contact times. Permethrin/malathion-resistant strains of Pediculus humanis capitis adults and eggs were directly exposed to Tenutex, and the vitality and hatchability, respectively, were evaluated. The anti-scabies study demonstrated that exposure for 6 or 24 h completely eradicated the parasite. The pediculicidal activity of Tenutex exhibited superior efficacy than standard treatment on resistant lice. The positive results obtained suggest that Tenutex® is a good treatment option, especially in drug resistance situations

Evaluación y caracterización de disulfiram como agente antimicrobiano de uso tópico

Actualment, la resistència antimicrobiana es considera una pandèmia silenciosa i una de les majors amenaces per a la salut a les quals s'enfronta la humanitat. En aquest context, la situació s'agreuja per la falta de descobriment de noves molècules actives. A més, en els últims anys, hi ha un creixent interès pels trastorns infectius de la pell i mucoses, per la seva alta prevalença i morbiditat. Per la qual cosa, és urgent reactivar la recerca i el desenvolupament de noves alternatives en aquestes àrees que permetin fer front a la farmacorresistència i superar les complexes característiques intrínseques a l' aplicació de fàrmacs en aquests teixits. En aquesta Tesi Doctoral, l'objectiu és avaluar i caracteritzar l'acció antimicrobiana de disulfiram en tres formulacions d'ús tòpic per abordar el problema de la resistència antimicrobiana des de diferents perspectives. En un primer estudi, s'ha determinat l'absorció transdèrmica i l'eficàcia in vitro contra sarna i polls d'una combinació de bencil benzoat i disulfiram (Tenutex®), producte comercial d'ús geogràficament restringit que podria ser una alternativa als insecticides estàndards. Es va utilitzar pell humana dermatomitzada per avaluar l'absorció dèrmica mitjançant un mètode validat de cromatografia líquida d'alt rendiment i es van utilitzar queratinòcits HEK001 per avaluar la citotoxicitat. Només un dels actius, el bencil benzoat, va poder permear la pell, però no va mostrar citotoxicitat en cap de les concentracions provades. L'eficàcia del producte es va assajar davant Psoroptes ovis a diferents temps de contacte i enfront de ceps resistents a la malatió d'adults i ous de Pediculus humanis capitis. L'estudi contra sarna va demostrar que l'exposició durant 6 o 24 h del producte eradicava completament el paràsit i, respecte l'activitat pediculicida, va mostrar una eficàcia superior a la del tractament estàndard en polls resistents. En el següent estudi, es va avaluar el reposicionament de disulfiram com a agent antibacterià per tractar infeccions de la pell. Aquest estudi va tenir com a objectiu determinar les característiques biofarmacèutiques del fàrmac i els efectes derivats de la seva aplicació tòpica. Les proves de susceptibilitat van mostrar activitat antibacteriana contra els bacteris grampositius Staphylococcus aureus i Streptococcus pyogenes. L'absorció dèrmica no va revelar permeació en la pell de porc i la quantificació del fàrmac retingut en teixit va demostrar concentracions en l'estrat còrni i epidermis suficients per tractar les infeccions. A més, es van realitzar anàlisis de micromatrius i citotoxicitat in vitro per comprendre millor el mecanisme del fàrmac. Aquests estudis van revelar la importància de disulfiram com a quelant d'ions metàl·lics. En el tercer estudi, a més de proposar el reposicionament de disulfiram com a agent antifúngic, es va desenvolupar i optimitzar formulacions alternatives per millorar l' eficàcia dels tractaments administrats per via vaginal. En aquest cas, es van elaborar gels mucoadhesius per a l'administració local del fàrmac. Aquestes formulacions estaven compostes de polietilenglicol i carragenat per millorar les propietats mucoadhesives i prolongar el temps de residència a la cavitat vaginal. Les proves de susceptibilitat van mostrar que disulfiram té activitat contra Candida albicans, Candida parapsilosis i Candida glabrata (Nakaseomyces glabratus). Es van caracteritzar les propietats fisicoquímiques dels gels i es van estudiar els perfils d'alliberament i permeació in vitro amb cel·les de Franz. A més, es va demostrar que la quantitat de fàrmac retingut en l'epiteli vaginal de porc era suficient per tractar la infecció. Atesa la situació d' emergència de resistència als antimicrobians, els resultats obtinguts en les tres publicacions científiques posen de manifest el potencial ús de disulfiram com a agent antimicrobià, demostrant que les formulacions proposades són una alternativa interessant per al tractament en infeccions cutanis i vaginals.Actualmente, la resistencia antimicrobiana se considera una pandemia silenciosa y una de las mayores amenazas para la salud a las que se enfrenta la humanidad. En este contexto, la situación se agrava por la falta de descubrimiento de nuevas moléculas activas. Además, en los últimos años, hay un creciente interés por los trastornos infectivos de la piel y mucosas, por su alta prevalencia y morbilidad. Por lo que, es urgente reactivar la investigación y el desarrollo de nuevas alternativas en estas áreas que permitan hacer frente a la farmacorresistencia y superar las complejas características intrínsecas a la aplicación de fármacos en estos tejidos. En la presente Tesis Doctoral, el objetivo es evaluar y caracterizar la acción antimicrobiana de disulfiram en tres formulaciones de uso tópico para abordar el problema de la resistencia antimicrobiana desde diferentes perspectivas. En un primer estudio, se ha determinado la absorción transdérmica y la eficacia in vitro contra sarna y piojos de una combinación de bencil benzoato y disulfiram (Tenutex®), producto comercial de uso geográficamente restringido que podría ser una alternativa a los insecticidas estándares. Se empleó piel humana dermatomizada para evaluar la absorción dérmica mediante un método validado de cromatografía líquida de alto rendimiento y se utilizaron queratinocitos HEK001 para evaluar la citotoxicidad. Sólo uno de los activos, el bencil benzoato, pudo permear la piel, pero no mostró citotoxicidad en ninguna de las concentraciones probadas. La eficacia del producto se ensayó frente a Psoroptes ovis a diferentes tiempos de contacto y frente a cepas resistentes a permetrina/malatión de adultos y huevos de Pediculus humanis capitis. El estudio contra sarna demostró que la exposición durante 6 o 24 h del producto erradicaba completamente el parásito y, respecto la actividad pediculicida, mostró una eficacia superior a la del tratamiento estándar en piojos resistentes. En el siguiente estudio, se evaluó el reposicionamiento de disulfiram como agente antibacteriano para tratar infecciones de la piel. Este estudio tuvo como objetivo determinar las características biofarmacéuticas del fármaco y los efectos derivados de su aplicación tópica. Las pruebas de susceptibilidad mostraron actividad antibacteriana contra las bacterias grampositivas Staphylococcus aureus y Streptococcus pyogenes. La absorción dérmica no reveló permeación en la piel de cerdo y la cuantificación del fármaco retenido en tejido demostró concentraciones en el estrato córneo y epidermis suficientes para tratar las infecciones. Además, se realizaron análisis de micromatrices y citotoxicidad in vitro para comprender mejor el mecanismo acción del fármaco. Estos estudios revelaron la importancia de disulfiram como quelante de iones metálicos. En el tercer estudio, además de proponer el reposicionamiento de disulfiram como agente antifúngico, se desarrolló y optimizó formulaciones alternativas para mejorar la eficacia de los tratamientos administrados por vía vaginal. En este caso, se elaboraron geles mucoadhesivos para la administración local del fármaco. Dichas formulaciones estaban compuestas de polietilenglicol y carragenato para mejorar las propiedades mucoadhesivas y prolongar el tiempo de residencia en la cavidad vaginal. Las pruebas de susceptibilidad mostraron que disulfiram tiene actividad contra Candida albicans, Candida parapsilosis y Candida glabrata (Nakaseomyces glabratus). Se caracterizaron las propiedades fisicoquímicas de los geles y se estudiaron los perfiles de liberación y permeación in vitro con celdas de Franz. Además, se demostró que la cantidad de fármaco retenido en el epitelio vaginal de cerdo era suficiente para tratar la infección. Dada la situación de emergencia de resistencia a los antimicrobianos, los resultados obtenidos en las tres publicaciones científicas ponen de manifiesto el potencial uso de disulfiram como agente antimicrobiana, demostrando que las formulaciones propuestas son una alternativa interesante para el tratamiento en infecciones cutáneos y vaginales.Currently, antimicrobial resistance is considered a silent pandemic and one of the greatest health threats facing humanity. In this context, the situation is aggravated by the lack of discovery of new active molecules. In addition, in recent years, there is a growing interest in infective disorders of the skin and mucous membranes, due to their high prevalence and morbidity. Therefore, it is urgent to reactivate research and development of new alternatives in these areas that allow us to address drug resistance and overcome the complex intrinsic characteristics to the application of drugs in these tissues. In this Doctoral Thesis, the objective is to evaluate and characterize the antimicrobial action of disulfiram in three formulations for topical use to address the problem of antimicrobial resistance from different perspectives. In a first study, the transdermal absorption and in vitro efficacy against scabies and lice of a combination of benzyl benzoate and disulfiram (Tenutex®), a commercial product of geographically restricted use that could be an alternative to standard insecticides, has been determined. Dermatomicized human skin was used to assess dermal absorption using a validated high-performance liquid chromatography method and HEK001 keratinocytes were used to evaluate cytotoxicity. Only one of the actives, benzyl benzoate, was able to permeate the skin, but it did not show cytotoxicity at any of the tested concentrations. The efficacy of the product was tested on Psoroptes ovis at different contact times and against adult permethrin/malathion resistant strains and eggs of Pediculus humanis capitis. The anti-scabies study demonstrated that exposure for 6 or 24 h of the product completely eradicated the parasite and, with respect to pediculicidal activity, it exhibited superior efficacy to that of standard treatment in resistant lice. In the following study, the repositioning of disulfiram as an antibacterial agent to treat skin infections was evaluated. This study aimed to determine the biopharmaceutical characteristics of the drug and the effects arising from its topical application. Susceptibility tests showed antibacterial activity against the Gram-positive bacteria Staphylococcus aureus and Streptococcus pyogenes. Dermal absorption revealed no permeation in pig skin and the quantification of the drug retained in tissue demonstrated concentrations in the stratum corneum and epidermis, enough to treat infections. In addition, micro-array and in vitro cytotoxicity analyses were performed to better understand the mechanism of action of the drug. These studies revealed the importance of disulfiram as a of metal ion chelator. In the third study, in addition to the repositioning of disulfiram as an antifungal agent, alternative formulations were developed and optimized to improve the efficacy of vaginally administered treatments. In this case, mucoadhesive gels were developed for local administration of the drug. These formulations were composed of polyethylene glycol and carrageenan to improve mucoadhesive properties and to prolong the residence time in the vaginal cavity. Susceptibility tests showed that disulfiram has activity against Candida albicans, Candida parapsilosis and Candida glabrata (Nakaseomyces glabratus). The physicochemical properties of the gels were characterized, and the release and permeation profiles were studied in vitro with Franz cells. In addition, it was shown that the amount of drug retained in the pig vaginal epithelium was sufficient to treat the infection. Given the emergency of antimicrobial resistance, the results obtained in the three publications highlight the potential use of disulfiram as an antimicrobial agent, demonstrating that the proposed formulations are an interesting alternative for the treatment of skin and vaginal infections

Repurposing Disulfiram as an Antimicrobial Agent in Topical Infections

Antimicrobial drugs applied topically offer several advantages. However, the widespread use of antibiotics has led to increasing antimicrobial resistance. One interesting approach in the drug discovery process is drug repurposing. Disulfiram, which was originally approved as an anti-alcoholism drug, offers an attractive alternative to treat topical multidrug resistance bacteria in skin human infections. This study aimed to evaluate the biopharmaceutical characteristics of the drug and the effects arising from its topical application in detail. Microdilution susceptibility testing showed antibacterial activity against Gram-positive bacteria Staphylococcus aureus and Streptococcus pyogenes. Dermal absorption revealed no permeation in pig skin. The quantification of the drug retained in pig skin demonstrated concentrations in the stratum corneum and epidermis, enough to treat skin infections. Moreover, in vitro cytotoxicity and micro-array analyses were performed to better understand the mechanism of action and revealed the importance of the drug as a metal ion chelator. Together, our findings suggest that disulfiram has the potential to be repurposed as an effective antibiotic to treat superficial human skin infections

Repurposing Disulfiram as an Antifungal Agent: Development of a New Disulfiram Vaginal Mucoadhesive Gel

Alternative formulations need to be developed to improve the efficacy of treatments administered via the vaginal route. Mucoadhesive gels with disulfiram, a molecule that was originally approved as an antialcoholism drug, offer an attractive alternative to treat vaginal candidiasis. The aim of the current study was to develop and optimize a mucoadhesive drug delivery system for the local administration of disulfiram. Such formulations were composed of polyethylene glycol and carrageenan to improve the mucoadhesive and mechanical properties and to prolong the residence time in the vaginal cavity. Microdilution susceptibility testing showed that these gels had antifungal activity against Candida albicans, Candida parapsilosis, and Nakaseomyces glabratus. The physicochemical properties of the gels were characterized, and the in vitro release and permeation profiles were investigated with vertical diffusion Franz cells. After quantification, it was determined that the amount of the drug retained in the pig vaginal epithelium was sufficient to treat candidiasis infection. Together, our findings suggest that mucoadhesive disulfiram gels have the potential to be an effective alternative treatment for vaginal candidiasis

Repurposing Disulfiram as an Antimicrobial Agent in Topical Infections

Antimicrobial drugs applied topically offer several advantages. However, the widespread use of antibiotics has led to increasing antimicrobial resistance. One interesting approach in the drug discovery process is drug repurposing. Disulfiram, which was originally approved as an anti-alcoholism drug, offers an attractive alternative to treat topical multidrug resistance bacteria in skin human infections. This study aimed to evaluate the biopharmaceutical characteristics of the drug and the effects arising from its topical application in detail. Microdilution susceptibility testing showed antibacterial activity against Gram-positive bacteria Staphylococcus aureus and Streptococcus pyogenes. Dermal absorption revealed no permeation in pig skin. The quantification of the drug retained in pig skin demonstrated concentrations in the stratum corneum and epidermis, enough to treat skin infections. Moreover, in vitro cytotoxicity and micro-array analyses were performed to better understand the mechanism of action and revealed the importance of the drug as a metal ion chelator. Together, our findings suggest that disulfiram has the potential to be repurposed as an effective antibiotic to treat superficial human skin infections