Desarrollo de nanocápsulas lipídicas como estrategia para facilitar el paso a través de la barrera hematoencefálica de fármacos que actúan a nivel del sistema nervioso central

Tesis de la Universidad Complutense de Madrid, Facultad de Farmacia, Departamento de Farmacia Galénica y Tecnología Alimentaria, leída el 17/09/2018. Tesis formato europeo (compendio de artículos)Diseases affecting the central nervous system (CNS) should be regarded as a major health challenge due to their steadily rising incidences and to the current lack of effective treatments given the hindrance to brain drug delivery imposed by the blood-brain barrier (BBB). Some of the described delivery strategies to circumvent the BBB such as the direct intracerebral administration and the artificial disruption of the tight junctions involve high risk of neurological damage. Hence, every effort is currently being devoted to achieving efficient transport across the brain endothelium with targeted drug carriers following minimally-invasive intravenous injection. In particular, nanomedicine is chiefly germane to the field of chemotherapy wherein dose availability at the target site cannot be enhanced by dose increase for fear of severe side effects. Since efficient brain targeting should not solely rely on passive targeting, brain active targeting of nanomedicines into the CNS is being explored...Las patologías que afectan al sistema nervioso central representan un desafío terapéutico por su incidencia creciente y la limitación del acceso a sistema nervioso central de la mayoría de fármacos administrados por vía sistémica por parte de la barrera hematoencefálica. Algunas de las estrategias para sortear esta barrera, incluyendo la administración intracerebral o la disrupción artificial de sus uniones estrechas, suponen un elevado riesgo de daño neurológico. Por ello, actualmente se persigue diseñar transportadores de fármacos capaces de atravesar de manera eficiente el endotelio cerebral tras su administración intravenosa. En concreto, la vectorización de antineoplásicos en nanotransportadores para el tratamiento de tumores cerebrales supondría un sustancial avance en terapéutica por la reducción de efectos secundarios derivados de su distribución sistémica. Dado que la distribución de transportadores a sistema nervioso central no puede depender en exclusiva de la vectorización pasiva, a fin de favorecer su paso a través de la barrera hematoencefálica, se está investigando la incorporación de distintos ligandos a estos sistemas. El objetivo global de esta tesis doctoral es diseñar, desarrollar y evaluar a nivel preclínico un nanotransportador lipídico capaz de atravesar la barrera hematoencefálica para vectorizar fármacos a nivel del sistema nervioso central tras una administración intravenosa. Este objetivo general se desglosa en tres objetivos específicos Estudiar los parámetros experimentales determinantes en la obtención de nanocápsulas lipídicas mediante el método térmico de inversión de fases para habilitar la producción de nanocápsulas lipídicas bajo demanda. Desarrollar una novedosa estrategia de funcionalización de nanocápsulas lipídicas con cannabidiol para favorecer su distribución a sistema nervioso central y evaluar su potencial in vitro e in vivo. Encapsular cannabidiol en el núcleo oleoso de las nanocápsulas y evaluar in vitro su eficacia como sistemas de liberación prolongada con actividad frente a la línea celular U373MG de glioblastoma humano. Asimismo, se persigue evaluar la estrategia de funcionalización con cannabidiol para potenciar la captación por células de glioma. En cuanto a los resultados, la obtención de nanocápsulas lipídicas con tamaños de partícula predeterminados para aumentar las posibilidades de éxito de tratamientos de patologías del sistema nervioso central puede conseguirse mediante el método térmico de inversión de fases, pues el diámetro volumen se ajusta a un modelo matemático en una variable (el cociente másico entre la fase interna oleosa y el tensioactivo). Este modelo es válido para nanocápsulas blancas y cargadas con fármaco. Además, una disminución de tamaño de las nanocápsulas lipídicas en el intervalo 20-60 nm incrementa en 2,5 y 1,6-2,5 veces su paso a través de la barrera hematoencefálica in vitro e in vivo, respectivamente. Por otra parte, la funcionalización de las nanocápsulas lipídicas con el fitocannabinoide cannabidiol aumenta el paso a través de barrera hematoencefálica 4,3 y 2,5 veces in vitro e in vivo, respectivamente. Una disminución de tamaño de las nanocápsulas lipídicas incrementa 3 veces la captación por células de glioma. Asimismo, el tamaño de las nanocápsulas lipídicas condiciona la liberación de fármacos: nanocápsulas de 20 nm cargadas con cannabidiol reducen invariablemente 3 veces los valores de concentración inhibitoria 50 en comparación con sus homólogas de 50 nm. Además, la funcionalización de nanocápsulas lipídicas con cannabidiol aumenta la captación por células de glioma 3,4 veces. Como conclusión, las nanocápsulas lipídicas, cargadas y funcionalizadas con cannabidiol, son prometedores candidatos para el tratamiento de gliomas con capacidad de vectorización a través de barrera hematoencefálica y de células de glioma. Su potencial terapéutico debe ser evaluado en modelos animales de glioma.Depto. de Farmacia Galénica y Tecnología AlimentariaFac. de FarmaciaTRUEunpu

Enhancing cannabidiol bioaccessibility using ionic liquid as emulsifier to produce nanosystems: Characterization of structures, cytotoxicity assessment, and in vitro digestion

The emulsifying potential of a biocompatible ionic liquid (IL) to produce lipid-based nanosystems developed to enhance the bioaccessibility of cannabidiol (CBD) was investigated. The IL (cholinium oleate) was evaluated at concentrations of 1 % and 2 % to produce nanoemulsions (NE-IL) and nanostructured lipid carriers (NLC-IL) loaded with CBD. The IL concentration of 1 % demonstrated to be sufficient to produce both NE-IL and NLC-IL with excellent stability properties, entrapment efficiency superior to 99 %, and CBD retention rate of 100 % during the storage period evaluated (i.e. 28 days at 25 °C). The in vitro digestion evaluation demonstrated that the NLC-IL provided a higher stability to the CBD, while the NE-IL improved the CBD bioaccessibility, which was mainly related to the composition of the lipid matrices used to obtain each nanosystem. Finally, it was observed that the CBD cytotoxicity was reduced when the compound was entrapped into both nanosystems.info:eu-repo/semantics/publishedVersio

Novel cannabinoid release system : encapsulation of a cannabidiol precursor into γ-cyclodextrin metal-organic frameworks

Altres ajuts: acord transformatiu CRUE-CSICAltres ajuts: Jorge Rodríguez-Martínez acknowledges Agència de Gestió d'Ajuts Universitaris i de Recerca from Generalitat de Catalunya (Sapin) for the FI-2018 fellowship.γ-Cyclodextrin-metal-organic frameworks (γ-CD-MOFs) are developed as a new promising and biocompatible material, which shows a great potential for drug delivery system (DDS) applications. γ-CD-MOFs were successfully synthesized using microwave-assisted technique from different potassium sources (KOH, KCl and KNO3). The encapsulation of olivetol (OLV) into these materials was investigated as an innovative model of DDS for cannabinoids. Loading of OLV in γ-CD-MOFs was performed by impregnation and co-crystallization methods. Scanning electron microscopy (SEM) and X-ray powder diffraction (XRPD) were employed to study the structural properties of γ-CD-MOF samples, showing the typical cubic crystals in case of KOH and trigonal morphologies in case of KCl and KNO3. Olivetol content was determined using UV-Vis spectrophotometry and its interaction with γ-CD-MOFs was investigated by Attenuated Total Reflection-Fourier Transform Infrared Spectroscopy analysis (ATR-FTIR). OLV content was significantly higher when KCl or KNO3 were employed in combination with a cocrystallization method, while the drug encapsulation using KOH and the impregnation method was really poor. For the first time, γ-CD-MOFs loaded with cannabinoids were developed and they could be considered a novel strategy as DDS of these compounds

Exosomal cannabidiol: A promising candidate for targeted oral delivery against breast cancer.

Cancer therapies are limited by lack of specificity, selectivity and numerous adverse effects. Repurposing potential anticancer compounds with appropriate drug carriers can help overcome these drawbacks. In our present study, we address the major obstacle for cannabidiol (CBD): its low oral bioavailability, using exosomes. Exosomes (Exo) are promising non-toxic and biocompatible drug delivery vehicles Our project aims are to address the research gap by formulating exosomal cannabidiol (ExoCBD): a tumor-targeted oral delivery system for the treatment of breast cancer. The targeting approach relies on the overexpression of folate receptors at the tumor site by using folic acid functionalized exosomes (FA-Exo). We loaded CBD onto the exosomes with 20 % drug load. The CBD-loaded exosomal formulations were well within the nanometric range and had an average size of 123 ± 2.4 nm. Triple-negative breast cancer (TNBC) cell lines – MDA-MB-231 and MDA-MB-231TR were more sensitive to CBD treatment as compared to MCF-7 and MCF-7TR. CBD exerted its anti-cancer effect by downregulating anti-apoptotic markers, anti-inflammatory maker at protein level. The ExoCBD formulation when administered orally, showed higher efficacy than the free drug (55 % vs 34%) against the orthotopic breast tumor growth. The efficacy was further enhanced to 68% with FA-ExoCBD. We also observed that ExoCBD showed higher levels of CBD as compared to free CBD in plasma, liver, and mammary pads upon oral administration of a single high dose. Thus, FA-ExoCBD can be further advanced as a cancer therapeutic considering its advantages such as low effective dose, with anti-inflammatory and pain-relieving effects

Cannabidiol as Self-Assembly Inducer for Anticancer Drug-Based Nanoparticles

Cannabidiol (CBD) is a biologically active compound present in the plants of the Cannabis family, used as anticonvulsant, anti-inflammatory, anti-anxiety, and more recently, anticancer drug. In this work, its use as a new self-assembly inducer in the formation of nanoparticles is validated. The target conjugates are characterized by the presence of different anticancer drugs (namely N-desacetyl thiocolchicine, podophyllotoxin, and paclitaxel) connected to CBD through a linker able to improve drug release. These nanoparticles are formed via solvent displacement method, resulting in monodisperse and stable structures having hydrodynamic diameters ranging from 160 to 400 nm. Their biological activity is evaluated on three human tumor cell lines (MSTO-211H, HT-29, and HepG2), obtaining GI(50) values in the low micromolar range. Further biological assays were carried out on MSTO-211H cells for the most effective NP 8B, confirming the involvement of paclitaxel in cytotoxicity and cell death mechanis

Cannabinoid Formulations and Delivery Systems: Current and Future Options to Treat Pain

The field of Cannabis sativa L. research for medical purposes has been rapidly advancing in recent decades and a growing body of evidence suggests that phytocannabinoids are beneficial for a range of conditions. At the same time impressing development has been observed for formulations and delivery systems expanding the potential use of cannabinoids as an effective medical therapy. The objective of this review is to present the most recent results from pharmaceutical companies and research groups investigating methods to improve cannabinoid bioavailability and to clearly establish its therapeutic efficacy, dose ranges, safety and also improve the patient compliance. Particular focus is the application of cannabinoids in pain treatment, describing the principal cannabinoids employed, the most promising delivery systems for each administration routes and updating the clinical evaluations. To offer the reader a wider view, this review discusses the formulation starting from galenic preparation up to nanotechnology approaches, showing advantages, limits, requirements needed. Furthermore, the most recent clinical data and meta-analysis for cannabinoids used in different pain management are summarized, evaluating their real effectiveness, in order also to spare opioids and improve patients’ quality of life. Promising evidence for pain treatments and for other important pathologies are also reviewed as likely future directions for cannabinoids formulations

Formulation Development, Preclinical Testing, and Primary Packaging Optimization for Cannabinoids and Other Therapeutics

Through the process of drug development, a molecule goes through discovery screening; lead selection and optimization, ADME testing, toxicity profiling, dosage form determination, preclinical testing in an in vitro and in vivo setup, folloby clinical research, FDA review and approval until eventually it is manufactured in the determined dosage form and reaches the patient. At every point through this process, scientists actively work towards a smoother transition and a quick and safe approval of the molecule towards the next step. The different chapters in this research would cover various phases of drug development; from discovery stage to fill-finish and primary container compatibility

The possible enhancement of colon cancer targeted photodynamic therapy when combined with Cannabidiol

Abstract: Colorectal cancer (CRC) remains the third most prevalent cancer and the fourth most common cause of cancer-related mortality (Mármol et al., 2017). In recent years, the incidence and mortality rate of CRC has been rampant in several developing countries (Benarba et al., 2018). Although it is feasible to combat some cancers, this does not hold true for CRC due to its asymptomatic nature in the early stages, hence it is often diagnosed after symptoms become noticeable (Mishra et al., 2013). There are several therapeutic methods available for CRC management, such as surgical excision, chemotherapy and radiotherapy (Hodgkinson et al., 2017). However, these therapeutic modalities have rarely yielded good prognosis in patients and have detrimental side effects (Mishra et al., 2013). Moreover, evolving resistance of cancer cells towards chemotherapeutic and biological drugs, as well as non-specific toxicity of these drugs on healthy tissues creates the need to explore other methods of CRC therapy (Hodgkinson et al., 2017)...M.Tech. (Biomedical Technology

Leukemia treatment: studies exploring bone marrow microenvironment, drug resistance and cannabidiol

The results described in this thesis highlight the role of human regenerative associated endothelial cells (hRECs) in the bone marrow microenvironment and hematopoiesis. By being resistant to 5-FU therapy, hRECs remain active in secreting factors enabling enhanced expansion of precursor HSC. A second part of this thesis provides novel insights into the role of specific influx and efflux transporters implicated in the efficacy/resistance of MTX and Ara-C. Finally, a third part described the results of in vitro experimental anti-leukemia therapeutics with cannabidiol, revealing double-edged effects: anti-leukemic effects at high doses of CBD and cell growth stimulatory effects at low doses of CBD. Together, these results may contribute to improve anti-leukemic chemotherapy efficacy and minimize toxicity