Los tratamientos considerados para el dolor y la inflamación han demostrado efectos colaterales, por ello, el descubrimiento e investigación de nuevos fármacos de origen natural son considerados como alternativa favorable. Los flavonoides comprenden un grupo de compuestos del tipo fenólico que abarcan una gran variedad de sustancias dentro de los Productos Naturales. Es por ello que en el presente trabajo se asilaron cuatro flavanonas isoprenílicas: 5, 7-dihidroxi-6-metil-8-prenilflavanona (I), 5, 7-dihidroxi-6-metil-8-prenil-4’-metoxi-flavanona (II), 5, 7-dihidroxi-6-prenilflavanona (III) y 5-hidroxi-7-metoxi-6-prenilflavanona (IV) a partir del extracto metanólico del las hojas de la especie vegetal Eysenhardtia platycarpa. A estas flavanonas se les evaluó su eficacia antiinflamatoria en un modelo de TPA en oreja de ratón, su capacidad antioxidante utilizando DPPH como radical libre y finalmente se hicieron pruebas de citotoxicidad frente a Artemia salina. Posteriormente, se elaboraron nanoformulaciones, en específico nanoemulsiones (NEs) y nanopartículas (NPs) a partir de las cuatro flavanonas Las NPs fueron del tipo polimérico preparadas mediante el método del desplazamiento del disolvente descrito por Fessi utilizando como polímero el PLGA 50/50 a la concentración de 1.5 mg/mL de principio activo. Estas NPs mostraron un tamaño de partícula medio de 156 - 202 nm con una carga negativa entre -25 and -30 mV. Los estudios de microscopía de transmisión electrónica (TEM) evidencian que las NPs tienen forma esférica obteniéndose de esta manera nanoesferas con un patrón denso uniformemente distribuido. Para el caso de las NEs se eleboraron empleando el método de pesada directa a una concentración de 0.5 % de principio activo. Los estudios de TEM y DLS (Dynamic light scattering) indican que se obtuvieron NEs del tipo W/O, de forma esférica y una estructura micelar de las gotas que oscilaban entre 60 y 70 nm; obteniéndose un menor tamaño de gotícula que en el caso de las NPs. A partir de las NPs y NEs se realizaron ensayos de liberación donde se encontró que los compuestos vehiculizados en nanopartículas liberan más rápido que en naoemulsiones; a las 2 h y a las 10 h. respectivamente y en ambos casos, las flavanonas (I) y (III) son las que mayor cantidad de fármaco liberan a esos tiempos. De los ensayos de permeación se observó que la constante de permeación y la cantidad retenida en piel se ven incrementados para las flavanonas (I), (II) y (IV) cuando son vehiculizadas en NPs. Con base a los resultados de la presente investigación se considera que la flavanona 5-hidroxi-7-metoxi-6-prenil flavanona (III) vehiculizada en nanoemulsión y nanopartículas poliméricas presenta los mejores resultados de permeación y actividad antiinflamatoria por lo que puede ser considerado un potencial agente alternativo antiinflamatorio de uso tópico. / The clinically used antiinflammatory drugs suffer from the disadvantage of side effects, though the discovery and research of new natural drugs are a better alternative. Flavonoids are a wide group of natural phenolic compounds. In the present research, some nanoformulations were elaborated, specifically nanoemulsions (NEs) and nanoparticles (NPs), from four isoprenylic flavanones: 5, 7-dihydroxy-6-metil-8-prenylflavanone (I), 5, 7-dihydroxy-6-metil-8-prenyl-4’-methoxy-flavanone (II), 5, 7-dihydroxy-6-prenylflavanone (III) y 5-hydroxy-7-methoxy-6-prenylflavanone (IV) for using them as topical antiinflamatory drugs. The NPs were polimerycal compounds prepared by the solvent displacement technique described by Fessi using the PLGA 50/50 as a polymer (Concentration of active: 1.5 mg/mL). These NPs showed an average particle size of 156-202 nm with a negative charge between -25 and -30 mV. Microscopical studies (TEM) reveal that the NPs have a spherical shape, getting then nanospheres with a dense pattern homogeneously distributed. In the case of the NEs, they were prepared using the direct weighing method at a 0.5 % concentration of active. The TEM and DLS (Dynamic light scattering) studies confirmed that all W/O NEs were obtained, in round shape and a micellar structure of the drops between 60 and 70 nm; whereas in the NPs droplets of smaller size were obtained. Release studies revealed that the NPs vehiculized compounds release rate is higher than the one in the NEs (at 2 and 10 hours, respectively) and in both cases, flavanones (I) and (III) are the ones with the highest amount of drug released. Permeation studies showed that the permeation constant and the retained amount in skin are increased in flavanones (I), (II) and (IV) when they are vehiculized in NPs. According to the present research results we confirm that flavanone 5-dihydroxy-6-methoxy-6-prenylflavanone (III) vehiculized in NEs and polymeric NPs shows the best permeation and anti-inflammatory activity results; therefore it may be considered a potential alternative anti-inflammatory topical agent.

El principal factor desencadenant de la neurodegeneració que caracteritza la malaltia d’Alzheimer (MA) és el pèptid β-amiloide (Aβ). La formació incrementada d’aquest pèptid i la seva agregació es troben a l’inici d’una cascada de processos neurotòxics que culminen amb una disfunció neuronal generalitzada i mort neuronal. Actualment, es troben en fase clínica diferents candidats a fàrmacs anti-Alzheimer denominats “modificadors de la malaltia” que van dirigits a inhibir la formació o l’agregació de l’Aβ. S’espera que aquests compostos puguin prevenir o frenar la neurodegeneració de la MA. L’arsenal terapèutic actual per al tractament de la MA continua estant dominat per un grup de fàrmacs que van estar desenvolupats per a tractar la seva simptomatologia, la qual apareix com a conseqüència de dèficits de diferents neurotransmissors, especialment d’acetilcolina (ACh), al sistema nerviós central. Concretament, quatre dels cinc fàrmacs actualment comercialitzats per al tractament de la MA són un grup d’inhibidors de l’enzim acetilcolinesterasa (AChE), enzim responsable de la hidròlisi de l’ACh. Recentment s’ha demostrat que l’AChE es pot unir a l’Aβ accelerant la seva agregació, augmentant-ne la neurotoxicitat i promovent el deteriorament cognitiu. A través del denominat lloc perifèric, el qual es troba a l’entrada d’una gorja catalítica d’uns 20 Å de longitud, al fons de la qual es troba el lloc catalític de l’enzim. Així, el bloqueig simultani amb ambdós llocs d’unió (actiu i perifèric) ha portat al desenvolupament dels denominats inhibidors d’AChE de lloc d’unió dual, els quals són candidats molt prometedors a fàrmacs anti-Alzheimer amb potencial per modificar positivament el curs de la malaltia. D’altra banda, degut al caràcter multifactorial de l’etiologia de la MA s’està començant a considerar el desenvolupament de nous compostos multipotents capaços d’interaccionar amb diferents dianes biològiques involucrades en la malaltia com a única via per a aconseguir incidir eficientment sobre la malaltia, com ara, l’estrès oxidatiu degut a la sobreproducció d’espècies reactives d’oxigen (ROS). S’ha demostrat que els compostos quelants d’ions metàl•lics són capaços d’inhibir la producció de ROS i s’està considerant la utilització d’antioxidants naturals contra processos en els quals està implicat l’estrès oxidatiu, com és el cas de la MA. / The main factor triggering the process of neurodegeneration characteristic of Alzheimer's disease (AD) is the so-called β-amyloid peptide (Aβ). The increased formation of this peptide and its aggregation are placed at the beginning of a cascade of neurotoxic processes culminating in widespread neuronal dysfunction and death. Currently, there are several "disease-modifying" anti-Alzheimer drug candidates in clinical trials aimed at inhibiting the formation or aggregation of Aβ. It is expected that these compounds can prevent or slow the neurodegeneration of AD. The current treatment available for the treatment of AD is still dominated by a group of drugs that were developed to treat its symptoms, which appear as a result of the decrease in the production of various neurotransmitters, particularly acetylcholine (ACh) at the central nervous system. Consequently, the use of cholinomimetic agents capable to compensate this deficit should reduce cholinergic symptoms of AD. In fact, four out of the five drugs currently marketed for the treatment of AD are a group of inhibitors of the enzyme acetylcholinesterase (AChE), the enzyme responsible for the hydrolysis of ACh at the synapses, which increase levels of the neurotransmitter, thus reducing the symptoms of AD. Recently, it has been demonstrated that the enzyme AChE can bind to Aβ accelerating its aggregation, increasing the neurotoxicity and promoting cognitive decline. The binding site of AChE to Aβ is the so-called peripheral site, which is located at the entrance of a catalytic gorge of about 20 Å length, at the bottom of which the catalytic site of the enzyme is placed, where the hydrolysis of ACh takes place. Thus, the blockade of the peripheral site of AChE should lead to an inhibition of Aβ aggregation and of the neurodegenerative cascade. In addition, the particular architecture of the enzyme where the active and peripheral sites are separated but close enough to allow a molecule to simultaneously interact with the two binding sites has led to the development of dual binding site AChE inhibitors, which are very promising anti-Alzheimer drug candidates expected to positively alter the course of disease. Moreover, due to the multifactorial nature of the etiology of AD, development of multipotent compounds capable of interacting with different biological targets involved in the disease is beginning to be considered as the only way to achieve efficient impact on the course of the disease. Aβ aggregation plays a crucial role in the pathogenesis of AD, inducing processes in the neurodegenerative cascade and oxidative stress due to overproduction of reactive oxygen species (ROS). It also appears that the Cu2+ and Fe3+ ions are capable of generating ROS, after binding to the Aβ For these reasons, the metal ions and oxidative stress are interesting targets to modulate by multipotent anti-Alzheimer drug candidates. Thus, it has been shown that the compounds capable of chelating metal ions are able to inhibit the production of ROS and the use of natural antioxidants is also being considered to block the oxidative stress involved in Aβ. The aim of the present PhD is to develop multipotent compounds capable to interact with different biological targets involved in AD, namely cholinesterases, Aβ and oxidative stress, in order to block its neurodegenerative cascade.

Tesis desarrollada en el Hospital de Bellvitge / El citomegalovirus humano (hCMV) es un herpes virus responsable de la mayoría de las infecciones virales oportunistas tras el trasplante de órgano sólido (TOS), con importantes efectos directos e indirectos que tienen consecuencias a largo plazo sobre la supervivencia del paciente y del injerto. El ganciclovir (GCV) IV ha sido la terapia de elección para el tratamiento de la infección y enfermedad por hCMV. Sin embargo, requiere administración endovenosa comportando hospitalización, necesidad de acceso vascular, elevados costes y complicaciones tipo infecciones y trombosis. El valganciclovir (VGC), un profármaco de ganciclovir desarrollado más recientemente, con una biodisponibilidad del 60% y buenos resultados preliminares para la profilaxis del hCMV podría ser una alternativa eficaz. Los objetivos del presente proyecto de tesis han sido, i) evaluar la eficacia y seguridad de un tratamiento secuencial basado en una inducción corta con GCV IV 5 mg/kg/12h (5 días), seguida de VGC oral 900 mg/12h(16 días); ii) evaluar la exposición al fármaco en ambas ocasiones (IV y oral); ii) desarrollar un modelo farmacocinético poblacional con NONMEM VI. En el estudio se incluyeron 21 pacientes receptores de un TOS (renal, hepático y cardíaco) con infección/ enfermedad por hCMV. Se realizaron determinaciones virológicas los días basal, 5, 15, 21, 30, 60, 90 y 180. Se obtuvieron muestras de sangre a distintos tiempos los días 5 y 15 del tratamiento y se cuantificaron las concentraciones séricas de GCV. Resultados: Todos los pacientes respondieron al tratamiento, con mejoría clínica y erradicación o disminución de la carga viral de hCMV al final del tratamiento. Concretamente, 14 de 21 pacientes (66,7%) presentaron erradicación viral, mientras que 7 de 21 (33.3%) disminuyeron pero persistieron con viremia. La carga viral basal presentó diferencias estadísticamente significativas entre los pacientes con erradicación viral el día 21 (33.250 CMV DNA copias (rango 3940-227000)) frente a los que no erradicaron (200.000 CMV DNA copias (rango 31.800-517.000)), (p = 0.025). Durante el seguimiento post-tratamiento, 3 de 21 pacientes presentaron recurrencia clínica de la infección por CMV, con un total de 4 episodios. Todos los efectos adversos observados pudieron manejarse sin discontinuar el tratamiento excepto en un paciente que presentó pancitopenia requiriendo discontinuación. La exposición a ganciclovir, estimada mediante el área bajo la curva de concentraciones séricas (AUC), obtenida tras la administración de valganciclovir fue 1,24 veces (intervalo de confianza 90% 1.03-1.41) superior a la obtenida tras ganciclovir IV, pero sin ser la diferencia estadísticamente significativa (p = 0.054). Tampoco se observó acumulación de ganciclovir, siendo los índices de acumulación calculados para cada régimen de dosificación inferiores a 2, sugiriendo que el valganciclovir oral podría substituir al ganciclovir IV desde el inicio del tratamiento. El comportamiento farmacocinético del ganciclovir fue descrito por un modelo abierto de dos compartimentos con cinética de absorción y eliminación de orden uno y tiempo de latencia. La función renal determinada por el aclaramiento de creatinina (CLcr), estimado mediante Cocrkroft-Gault, fue la covariable más influyente en el aclaramiento del fármaco (CL). Los parámetros farmacocinéticos finales fueron: CL= 7,49 (CLCR/57) L / h (57: valor medio de CLcr de la población); volúmenes de distribución central y periférico, V2 = 31,9 L y V3 = 32,0 L, el aclaramiento intercompartimental CLD1 = 10,2 L / h, constante de velocidad de absorción de primer orden ka = 0.895 h-1; biodisponibilidad F = 0,825 y tiempo de latencia LT = 0.382 h. La validación interna del modelo demostró su capacidad predictiva. El modelo desarrollado permitió evaluar la adecuación de las dosis recomendadas por el fabricante ajustadas a CLcr para evitar sobre o infraexposición de ganciclovir, asumiendo que la viremia puede ser suprimida durante la profilaxis con una exposición de 40-50 (μg/ml)•h. / Human Cytomegalovirus (hCMV) is a herpesvirus, very common opportunistic pathogen after solid organ transplantation (SOT). Early treatment is required to avoid serious direct and indirect effects that can affect long term patient and graft survival. Ganciclovir (GCV) given orally or intravenously (IV) has been the first line agent for prevention and treatment of human Cytomegalovirus (hCMV) infection and disease in solid organ transplantation (SOT). The introduction of oral valganciclovir(VGC), with higher bioavailability than oral ganciclovir, has demonstrated to be a suitable approach in hCMV profilaxis therapy towards outpatient oral therapy. The aims of the present study were, i) to evaluate efficacy and safety of a short sequential treatment based on GCV IV (5 mg/kg twice daily) for 5 days followed by VGC oral (900 mg twice daily) for 16 days; ii) to evaluate the exposures achieved after both occasions; iii) to develop a population pharmacokinetic model with NONMEM VI. Twenty-one SOT patients with Cytomegalovirus infection/disease were included and followed until day 180. Pharmacodynamic assessments showed viral load eradication in 66.7% of patients, on day 21 (end of treatment), mainly in patients not at risk for primary CMV exposure. Mean exposure value to ganciclovir after oral valganciclovir was 1.24 times (90% confidence interval 1.03-1.41) that found after intravenous ganciclovir, so confirms that oral valganciclovir is an useful alternative to intravenous ganciclovir, with the benefit of increased convenience to the patient. A two-open compartment model best described the PK of GCV. The final pharmacokinetic parameters were: ganciclovir clearance CL = 7.49 (CLCR/57) L/h (57: mean population value of CLCR); central and peripheral distribution volumes, V2 = 31.9 L and V3 = 32.0 L; intercompartmental clearance CLD1 = 10.2 L/h; first-order absorption rate constant ka = 0.895 h-1; bioavailability F = 0.825 and lag time LT = 0.382 h. The renal function given by creatinine clearance (CLCR) was the most influential covariate in CL. After internal validation, the model was used to evaluate the adequacy of the manufacturer’s recommended CLCR adjusted doses to prevent under- or over-exposure, assuming that viraemia eradication can be suppressed during prophylaxis with exposures of 40–50 (g/mL)•h (Whiltshire and col ,2005).

La Tesi Doctoral s’emmarca dins del projecte de recerca adreçat al estudi de la química de sistemes moleculars basats en sals d’imidazoli, que ve desenvolupant el nostre grup en els darrers anys. En concret la present Tesi Doctoral s’ha proposat una alternativa als mètodes clàssics, poc estudiada fins ara en líquids iònics, pel procés de bescanvi d’anió. La metodologia desenvolupada és senzilla d’aplicar i eficient donat que millora els rendiments descrits en la literatura, a vegades de forma substancial, proporcionant parells iònics lliures d’halur. El mètode basat en l’ús de reïnes de bescanvi iònic, permet obtenir sals de cations orgànics amb l’anió objectiu seguint un procés on es poden diferenciar dues parts. La primera correspon a la carrega de la reïna amb l’anió seleccionat [AER (forma A¯)], on s’han estudiat diferents vies com a font de l’anió i diversos solvents de càrrega segons el seu caràcter més o menys hidròfil. La segona correspon al bescanvi de l’anió halur examinant diferents solvents i mescles segons les característiques dels productes. A la senzillesa del protocol, s’hi afegeix el fet de que es minimitza la formació de productes secundaris tòxics, i la reïna utilitzada pel bescanvi és reciclable, amb el conseqüent benefici medi ambiental, entrant dins l’àrea de la química sostenible. El mètode ha estat aplicat amb èxit al bescanvi del contraanió en sals d’imidazoli, benzimidazoli, piridini i triazoli, així com en sals d’amoni quaternàries, amb propietats com a líquids iònics. La selecció de l’anió permet modular les seves propietats fisico-químiques, obtenir ILs per tasques especifiques, i disminuir la seva ecotoxicitat, entre altres possibilitats. La metodologia s’ha estès amb resultats excel•lents a sensors i receptors d’anions basats en sals d’imidazoli, monocatiònics i dicatiònics de forma que es facilita l’accés a compostos que podran ser útils per a l’estudi de propietats en l’àrea del reconeixement molecular d’anions, el que obre noves perspectives per a l’estudi d’aquests sistemes. Per altre part poden preparar-se sals d’ingredients farmacèutics actius (APIs), estratègia que pot permetre la modulació de les seves propietats proporcionant millores que incloguin el control de la solubilitat, la biodisponibilitat o bioactivitat, l’estabilitat, l'eliminació de polimorfisme, noves formes d’alliberament, o fins i tot permetre l’administració de còctels de fàrmacs personalitzats. Així, el mètode s’ha aplicat en la preparació de sals àcids arilpropiònics amb propietats antiinflamatòries en forma de líquids iònics, accedint als corresponents arilpropionats d’1-butil-3-metilimidazoli amb els que s’ha iniciat l’estudi del perfil d’alliberament del fàrmac. En la darrera part de la tesi, i dins de l’àrea del reconeixement molecular s’ha estudiat el comportament com a receptor abiòtic d’anions de [14]heterofans mitjançant estudis de complexació de RMN de 1H, observant que en CD3CN presenta afinitat pels anions clorur i acetat segons un model d’estequiometria 1:1 amb un valor de Ka (constant d’associació) proper al 104 mol–1 per l’anió acetat. A més a més, els resultats obtinguts en DMSO d6 indiquen una considerable afinitat pel anió H2PO4¯ si bé l’estudi resta pendent de completar / The PhD thesis is part of a research project aimed at studying the chemistry of imidazolium molecular systems, which our group has been developing in recent years. Initially, the current work was focused on the development of a simple and efficient protocol to obtain a halide exchange with a broad range of organic and inorganic anions in imidazolium ionic liquids (ILs) using an anion exchange resin (AER) as an alternative to the classical methods. This approach involves two steps: a) loading the anion in the AER, with different anions of variable hydrophilic character exploring sources and solvents and b) performing the halide exchange with the most suitable solvent mixture to obtain the new halide-free ion pair. Besides the advantage of this simplicity, this protocol minimized the formation of toxic byproducts, essential for sustainable chemistry, as well as featuring a recyclable AER. The AER (A¯ form) method has been applied to swap halides for assorted anions in several quaternary heteroaromatic salts that include imidazolium, benzimidazolium, pyridinium and triazolium salts, and quaternary ammonium salts, providing the halide-free ion pairs in excellent to quantitative yields. The methodology has been extended with excellent results to anion receptors and sensors based on monocationic and dicationic imidazolium salts. Thus, access has been facilitated to potentially useful compounds in the area of anion molecular recognition, which opens new perspectives for the study of these systems. Additionally, the developed protocol was successfully extended to prepare salts from active pharmaceutical ingredients (APIs), modifying their properties. An imidazolium cation was combined with the anion of an anti-inflamatory arylpropionic acid to obtain the ionic liquid salt. A study of the release of API from hyaluronan-based hydrogels as drug delivery system is currently underway. In the final part of the thesis, specifically in the area of molecular recognition, we have explored the behaviour of [14]heterophanes as abiotic receptors using 1H-NMR techniques to study their anion binding properties. As an example, in CD3CN, the acetate anion showed a Ka value of nearly 104 mol–1. The results obtained open perspectives for further work in this field.

DE LA TESI:Una visió unificada de possibles estructures heterocícliques originals posa de manifest que les betaïnes formen un conjunt d'entitats químiques amb un elevat carácter dipolar i baix pes molecular <96CHC, 96CHC(3)3>. L'atractiu tant des d'un punt de vista bàsic de les betaïnes heterocícliques com el de les seves potencials aplicacions, es deuen fonamentalment a la seva naturalesa dipolar, que té una influència determinant en la seva química.Des de fa anys, el nostre grup d'investigació ha centrat l'atenció en l'estudi de les betaïnes heterocícliques d'azolat azoli(azini) amb una diferents agrupaments interanulars 1 i els seus precursors immediats 2 <94AHC(60)197, 96FE8381, 99FE8297>. Un dels aspectes que estem interesats en desenvolupar dins el camp de l'estudi de les betaïnes heterocícliques 1 i els derivats protonats 2 es aquell relacionat amb les potencials propietats biològiques d'aquest tipus de molècules. Les betaïnes heterocícliques 1 poden donar lIoc a una investigació en diferents camps farmacològics, com per exemple l'estudi d'interaccions amb la colina acetiltransferasa (ChAT) i amb l'acetilcolinesterasa (AChE). Entre l'àmplia i variada gamma d'activitats biològiques que presenten les sals quaternàries de piridini, diverses sèries han mostrat ésser inhibidors enzimàtics específics.Per altra banda, destaca l'interès d'aquestes betaïnes 1 com a substrats per a l'obtenció de cristalls líquids i materials orgànics amb propietats en òptica no-lineal <94AHC(60)197, 94JOC644, 96FES381, 99FES297> . Recentment, les investigacions dutes a terme per un grup d'investigació japonès corroboren que diferents tipus de betaïnes heterocícliques 1 presenten propietats excepcionals en òptica no-lineal, tant des d'un punt de vista teòric <96JA470S> com experimental <98MI1193>.A fi d'aprofundir en la comprensió de la química de les betaïnes heterocícliques 1, es considerà la seva inserció com a blocs fonamentals en sistemes macrocíclics, fet que permetrà endinsar-se en el camp del reconeixement molecular. La dificultat resideix en la selecció dels models de receptors. En aquest sentit, l'inexhaurible diversitat d'arquitectures moleculars en els sistemes macrocíclics -naturals i sintètics- podria permetre el disseny de nous compostos les característiques químiques dels quals influeixen de forma determinant sobre les seves propietats biològiques i físiques <96AG(E)1154, 96CSC(1), 96CSC(2». Els ciclofans, fans i heterofans, junt amb diferents tipus de calixarens representen una àmplia selecció de molècules i formes <96CSC(1), 96CSC(2» i constitueixen una font d'inspiració pel disseny d'originals receptors sintètics, però cap d'aquestes arquitectures moleculars està relacionada amb els [1 n]heterofans quadrupolars 4-6.Els components anulars presents en els heterofans són normalment subunitats heteroaromàtiques no carregades, i en els pocs casos en els quals soporten una càrrega, corresponen a nuclis quaternaris de piridini <88AG(E)1547, 91JA7033, 94AC2082, 96CSC(1), 96CSC(2» i azoli <99T2327>. Per una altra part, s'han descrit alguns ciclofans amb l'habilitat de reconèixer ions, encara que pocs d'ells presenten la capacitat de complexar anions <96RTC37, 97CRV1609>, malgrat el creixent interés en el reconeixement molecular d'anions <97MI1 >.Des de que es va sintetitzar l'[1(4)]azolofà quadrupolar 4 <95CC1239, 95SL757, 96T15171>, una part de la nostra recerca sobre nous substrats orgànics que incorporen subunitats betaíniques està relacionada amb arquitectures macrocícliques quadrupolars <99FES297>. Els heterofans que contenen betaïnes heterocícliques com subunitats emergeixen com una nova família d'[1(n)]heterofans i la seva natura dipolar o quadrupolar confereix propietats que tenen una clara incidència en la formació de complexos receptor-substrat.La informació obtinguda de les estructures en estat sòlid de l'[1(4) ]metaheterofà quadrupolar 7 <95CC1239>, l'[1(4)] (meta-orto)(2)heterofà quadrupolar 8 <96T15189> i l'[1(4)]]metaheterofà tripolar 9 <95CC1239> és de gran rellevància per avaluar la seva capacitat per actuar com receptors abiòtics Els receptors macrocíclics presenten normalment una conformació preorganitzada, però aquest no és un requisit essencial per assolir una complexació forta i selectiva. Així, en un estudi parat·lel realitzat, s'han sintetitzat diverses betaïnes heterocícliques de cadena oberta <97MI2>, on les subunitats heteroaromàtiques amb càrrega estan separades per llargues cadenes alifàtiques, les quals presenten la posibilidad d'actuar com pinces moleculars segons un mecanisme d'ajust induït.El disseny i generació de nous sistemes moleculars que incorporin betaïnes com a blocs fonamentals brinda un excel·lent potencial investigador. Les betaïnes, que poden actuar com a pinces dipolars, es poden estudiar tant des d'un punt de vista estructural com biològic. Pel cas dels heterofans dicatiònics i quadrupolars, amb diferents tamanys de cavitat i forma, són molècules prototipus per estudiar les seves propietats estructurals com la seva capacitant complexant (reconeixement molecular).En resum, els objectius de la present Memòria s'han centrat en tres camps. - Com a extensió lògica del treball previ realitzat al nostre laboratori <94AHC(60)197, 96FE8381, 99FE8297> es considerà, en primer lloc la síntesi, l'estudi estructural i l'avaluació com a receptors abiòtics d'anions dels macrocicles [14lmetaimidazoliofans 10·2X (Capítols 2 i 3 respectivament).- D'altra banda, el segon objectiu de la present Tesi Doctoral ha estat l'estudi de la reactivitat enfront d'electròfils -N-alquilació- dels [1(4)]heterofans quadrupolars 7 que incorporen a la seva molècula betaïnes d'1,2,4-triazolat metilenimidazoli i la caracterització estructural dels [14lmetaazolofans dicatiònics 11·2X i 12·2X resultants (Capítol 4).- Finalment, el darrer objectiu de la present Tesi Doctoral ha estat la síntesi i estudi estructural dels sistemes macrocíclics quadrupolars del tipus [1(6]heterofà 5 i [1(8)]heterofà 6.En el cas dels [1(6)]heterofans quadrupolars, les molècules diana 13 incorporen betaïnes d'1,2,4-triazolat metilenimidazoli com a subunitats. Els seus precursors immediats són els compostos dicatiònics 14·2X (Capítol 5).En el cas dels [1a]heterofans quadrupolars, les molècules diana 15 també incorporen betaïnes d'1,2,4-triazolat metilenimidazoli com a subunitats i els seus precursors immediats són els compostos dicatiònics 16·2X (Capítol 5).

El punt de partida de la present Tesi Doctoral se situa entorn a l'estudi d'estructures basades en (Z)-estilbens: disseny, síntesi i examen de quimioteques dirigides a l'obtenció de nous compostos bioactius a nivell de sistema nerviós central (SNC), en el marc d'una col·laboració amb la indústria farmacèutica. Així, el projecte es va iniciar amb una recerca bibliogràfica per la selecció del scaffold, la qual ens posà de manifest com els (Z)-arilmetilidenindens constituïen un conjunt de compostos amb diferents perfils farmacològics.Valorant aquests antecedents, la dificultat radicava en l'elecció de l'estructura prototipus. Els (Z) arilmetilidenindens podien ésser un inici ja que el fragment estructural cis estilbè s'integra en el scaffold. A més, la química de sistemes basats en indens i l'estudi de les seves propietats biològiques estaven menys explorats que la d'altres famílies de compostos, la qual cosa permetria desenvolupar temes específics d'investigació amb probabilitat raonable d'èxit.Concretament, se sintetitzaren els cis-indens I i les seves formes reduïdes II, basats en una selecció estructural, a partir de les indolilsulfonamides per substitució de l'indol per l'indè, i que incorporaven el grup sulfonamido en la posició 5. L'accés a les indenilsulfonamides II es portà a terme inicialment mitjançant una via específica, de cinc etapes, on els àcids (sulfonilamino)indenilacètics eren els intermedis claus, els quals es prepararen, després d'examinar diferents alternatives sintètiques, per reacció aldòlica.Els assajos d'afinitats pel receptor 5-HT6 mostraren valors diferents, sent moderats pels compostos I i augmentant de forma important per les indenilsulfonamides II. A més, cal destacar la funcionalitat agonista d'aquests derivats, ja que en els darrers anys, la majoria de lligands descoberts per aquest receptor havien estat identificats com antagonistes. Per aquesta raó, i tenint en compte que els canvis responsables de l'augment d'afinitat enfront el receptor 5-HT6 venien limitats per la química de l'indè, ens plantejàrem la síntesi d'altres estructures mitjançant una ruta més general de quatre etapes, on la sulfonilació es duia en la darrera etapa, sent les inden-5-amines els precursors de nous lligands II diversificats en la posició 5.Les noves indenilsulfonamides II exhibiren afinitats variables pel receptor 5 HT6, destacant dos compostos que s'identificaren com a potents agonistes selectius amb excel·lents perfils farmacològics in vitro (Ki &#8805; 4,5 nM).Alhora, s'ha estudiat la influència de la cadena N,N-dimetilaminoetil de la posició 3 sobre l'indè, amb a la preparació d'indenilsulfonamides restringides III i derivats simplificats, en base als requeriments estructurals establerts prèviament per augmentar l'afinitat dels lligands pel receptor 5 HT6. La síntesi d'aquests compostos es portà a terme mitjançant una via de tres etapes, a partir de les indanones convenientment substituïdes per preparar les indenamines precursores, aplicant la reacció aldòlica anteriorment emprada, adaptada a les lactames.Aquestes indenilsulfonamides restringides III exhibiren elevades afinitats pel receptor 5-HT6 actuant com antagonistes, tot i que amb potències moderades a nivell micromolar. D'altra banda, l'absència de la amina bàsica en els compostos estructuralment simplicats no implicava pèrdua d'afinitat, indicant com els grups N,N aminoetil o l'amina restringida sobre la posició 3 de l'indè no eren necessàris per mantenir l'afinitat pel receptor.Finalment, també s'ha portat a terme el canvi de la cadena aminoetílica per un grup guanilhidrazona rígid, que ens ha permès la identificació de les indanilguanilhidrazones sulfonamides IV com a potents i selectius antagonistes del receptor 5-HT6 amb elevades afinitats, a nivell nanomolar.Aquest conjunt de nous lligands basats en indens podrien ser útils eines d'elucidació d'aspectes funcionals i usos terapèutics d'altres lligands del receptor 5-HT6 en el tractament de malalties del SNC tals com l'ansietat, depressió o altres desordres mentals. Així, com l'interès que representen els agonistes enfront alteracions alimentàries tals com l'obesitat o la bulímia. / The background of this thesis is the study of structures based on (Z)-stilbenes: design, synthesis and testing of libraries designed to obtain new bioactive compounds in central nervous system, in collaboration with the pharmaceutical industry. So the structure proptotype, the (Z)-arylmethylideneindenes were chosen as the structural fragment cis-stilbenes integrated into the scaffold could form a set of compounds with different pharmacological profiles. In addition, chemical-based systems indenes such as I and the study of their biological properties were less explored than other families of compounds, which permit the development of specific topics of research with responsible likelihood of success.By developing an indole-to-indene core change, we have synthesized the cis indenes I and their reduced forms II, which incorporate the sulfonamido group in 5 position. These compounds have presented affinity for serotonin 5-HT6 receptor (Ki &#8805; 20 nM). The novel indenylsulfonamides II exhibited variable binding affinities for the 5-HT6 receptor and the in vitro profiles of the most active compounds revealed them to be selective 5 HT6 receptor agonists in the low-nanomolar range (Ki &#8805; 4.5 nM).Moreover, we have studied the influence of the N,N-dimethylaminoethyl side chain on the indene 3-position. Preparation of a few examples of ring-constrained III type indenylsulfonamides and the structurally simplified counterparts, based on previously established structural requirements for enhancing the affinity of indene based ligands towards the 5-HT6 receptor, gave access compounds which exhibited a high binding affinity and functioned as antagonists, although with moderate potency at the micromolar level. Finally, changing the N,N-(dimethylamino)ethyl side chain for a conformationally rigid guanylhydrazone moiety on the indene 3 position led to the identification of novel indanylguanylhydrazone sulfonamides IV with excellent binding affinities and an antagonistic response at the 5-HT6 receptor in the nanomolar range.The ensemble of indene-based frameworks constituted by the (aminoethyl)indenylsulfonamide agonists II, the conformationally rigid antagonists III and the structurally simplified counterparts, as well as the indanylsulfonamide guanylhydrazone antagonists IV (see Figure 1) resulted in a range of compounds which may be useful biological tools for the fundamental understanding of the 5-HT6 receptor.

Tesis por compendio de publicaciones / Objetivo: Explorar las relaciones entre las experiencias de los pacientes y sus evaluaciones del servicio en centros de rehabilitación. Métodos: Encuesta transversal multicéntrica usando cuestionario auto-administrado. Participaron 465 pacientes (90% contactados), edad media 39,4 (DE=11,9). El cuestionario incluyó medidas de experiencias y evaluaciones (satisfacción, calidad percibida y confianza). Resultados: Tanto el grado de calidad percibida como de confianza con la profesionalidad del fisioterapeuta mostraron buena correlación con la satisfacción (r=0,72 y r=0,672). Tres modelos de regresión indicaron que las experiencias fueron determinantes negativos en esas evaluaciones. Un 25% de pacientes con alta satisfacción y calidad percibida tuvieron muy baja cantidad de problemas. Sin embargo, muchos pacientes con alta satisfacción, calidad percibida y confianza manifestaron problemas de calidad. Conclusión: Satisfacción, calidad percibida y confianza son positivas y consistentes entre sí, pero proporcionan una medida limitada y optimista de la calidad asistencial: son positivas aún cuando algunas experiencias son de calidad mejorable. / Objective: To explore the relationships between patient experiences and their evalutions of service in outpatient rehabilitation centers. Methods: Cross-sectional, self-reported survey. Four hundred and sixty-five patients (response rate 90%). Mean age 39,4 (SD=11,9). Questionnaire measured patient experiences and overall evaluations (satisfaction, perceived service quality and trust). Results: Perceived service quality and trust in professionalism showed good correlation with satisfaction (r=0,72 y r=0,672). Three regression models showed experiences as negative determinants on those evaluations. Only 25% of participants with high satisfaction and perceived service quality had low amount of problems. However, many patients with high satisfaction, perceived service quality and trust showed quality problems. Conclusion: Satisfaction, perceived quality and trust are positive and consistent between them, but show a limited and optimistic picture of quality of care: they are positive while some experiences are improvable.

La medición de la adherencia es un requisito básico tanto en la práctica clínica como en la investigación. En el ámbito clínico se ha señalado que es importante evaluar y monitorizar la adherencia más que asumir que el consejo es suficiente o que los pacientes que son adherentes durante las sesiones lo son también en sus domicilios. En el ámbito de la investigación la medición de la adherencia es útil tanto para determinar el nivel o prevalencia como para su relación con otro tipo de variables que se quieran asociar, tales como sus beneficios, sus determinantes o las estrategias para su mejora. La medición de la adherencia terapéutica en los programas de fisioterapia es compleja porque habitualmente requieren diferentes actividades. Se ha señalado que la adherencia no puede ser medida de forma global sino que su medida necesita ser multifacética de acuerdo a cada uno de los comportamientos implicados. Objetivos: 1) Describir la frecuencia en la que los pacientes en fase post-aguda reciben durante la asistencia a Fisioterapia consejos que implican la realización de ejercicios domiciliarios específicos para su problema, ejercicios de acondicionamiento general y el área del automanejo del dolor. 2) Describir la adherencia existente al final del periodo de tratamiento de Fisioterapia a diferentes áreas de consejos proporcionados a los pacientes en fase post-aguda. 3) Analizar la adherencia a diferentes consejos de la Fisioterapia de los pacientes en fase post-aguda según el perfil del paciente, sus experiencias y su juicio global sobre la atención proporcionada. 4) Explorar y determinar la importancia relativa de diferentes factores explicativos a la adherencia a diferentes consejos de la Fisioterapia en el ámbito de la atención al paciente post-agudo. Material y Métodos: Se realizó un estudio transversal mediante cuestionario autocumplimentado por el paciente que recibe atención de fisioterapia por problemas músculoesqueleticos. Resultados y Discusión: El estudio se desarrolló con 516 sujetos, obteniendo 465 respuestas al cuestionario. Un alto porcentaje de pacientes recibieron consejo en relación a ejercicios específicos para su problema y medidas para el automanejo del dolor (88% y 83.1% respectivamente). Los consejos relativos al ejercicio aeróbico (45.9%) fueron recibidos por un menor porcentaje de pacientes. En cuanto al cumplimiento de las recomendaciones o adherencia por parte del paciente podemos señalar que es muy alta para los consejos relacionados al ejercicio aeróbico y a las medidas para el automanejo del dolor. Conclusiones: 1) Los fisioterapeutas suelen aconsejar el automanejo del dolor y la realización de ejercicios domiciliarios a la mayoría de sus pacientes post-agudos, y la realización de actividades aeróbicas a la mitad de ellos aproximadamente. 2) La adherencia al consejo fisioterápico es un problema en nuestro contexto cuya magnitud varía en función del tipo de consejo. Es un problema muy frecuente en el caso de los ejercicios domiciliarios específicos y, afortunadamente, menos frecuente para los ejercicios aeróbicos y automanejo del dolor. 3) No existe un perfil demográfico o clínico asociado a la adherencia a los ejercicios incluidos en este estudio. Por el contrario, el sexo femenino es un determinante de la adherencia a los consejos para el automanejo del dolor. 4) Los fisioterapeutas pueden incrementar la adherencia a los ejercicios domiciliarios específicos y el automanejo del dolor evitando experiencias problemáticas con la información y educación, y el apoyo emocional que prestan a sus pacientes. 5) Facilitar que los pacientes valoren el servicio recibido como de buena calidad o altamente satisfactorio puede favorecer respectivamente la adherencia a los ejercicios domiciliarios específicos y aeróbicos. / Measuring adherence is a basic requirement in both clinical practice and research . In clinical settings has indicated that it is important to assess and monitor adherence rather than assume that the advice is sufficient or that patients are adherent during sessions are also at home. In the research field of measuring adherence is useful for determining the level or prevalence as to its relationship with other variables that want to associate , such as benefits , its determinants and strategies for improvement. Measuring adherence in physiotherapy programs is complex because they usually require different activities. It has been suggested that adherence can not be measured global but multifaceted needs to be tailored according to each of the behaviors involved. Objectives: 1) Describe the frequency that patients in post- acute phase received while attending physiotherapy tips involve making specific home exercises for their problem , general conditioning exercises and pain self-management area . 2) Describe the adherence at the end of the physiotherapy treatment period to different areas of advice given to patients in post- acute phase . 3) Assess adherence to different physiotherapy advices of patients in post- acute phase according to the patient's profile , their experiences and their overall assessment of the care provided . 4) Explore and determine the relative importance of different factors explaining the different adherence to physiotherapy advices in the field of patient care post- acute . Material and Methods : We performed a cross-sectional study using self-administered questionnaire for the patient receiving physiotherapy care for musculoskeletal problems. Results and Discussion: The study was conducted with 516 subjects , obtaining 465 responses to the questionnaire. A high percentage of patients received advice in relation to specific exercises for your problem and measures for self-management of pain ( 88% and 83.1 % respectively). The advice relating to aerobic exercise (45.9 % ) were received by a smaller percentage of patients. Regarding compliance with recommendations or adherence by the patient can point which is very high for aerobic exercise -related advice and measures for self- pain . Conclusions : 1) Physical therapists often advise pain self-management and home-based exercises most of its post- acute patients , and aerobic activities to approximately half of them . 2) Adherence to physiotherapy advice is a problem in our context whose magnitude varies depending on the kind of advice . It is a very common problem in the case of specific home exercises and fortunately less common for aerobic exercise and self-management of pain. 3 ) There is no demographic or clinical profile associated with adherence to the exercises included in this study. By contrast , female gender is a determinant of adherence to self-management tips for pain . 4) Physical therapists can increase adherence to specific home exercises and avoiding pain self-management of problematic experiences with information and education , and emotional support they provide to their patients. 5) Give that patients value the service they received as good quality or highly satisfactory respectively may promote adherence to specific home exercises and aerobics .

Las proteínas de membrana constituyen entre un 25 y un 30 % de las proteínas presentes en las células, pudiendo estar implicadas, en algunos casos, en mecanismos de resistencia a determinados fármacos. Es por ello de gran interés el estudio de estas proteínas en modelos biomiméticos.El conocimiento existente sobre los fenómenos implicados en el proceso de reconstitución de proteínas transmembranarias en liposomas es limitado. Dicho proceso es fundamentalmente empírico, e implica la utilización de liposomas y surfactantes, así como proteínas con un alto grado de pureza. Estas condiciones son un requisito imprescindible para poder realizar estudios de integración de las proteínas, de su disposición en la bicapa y de su función biológica.La aplicación de técnicas nanométricas, como la microscopía de fuerza atómica (AFM), así como determinados métodos de fluorescencia (cálculo del incremento de potencial electrostático de superficie y anisotropía de la fluorescencia), permiten el estudio del proceso de reconstitución de proteínas.La aplicación de la AFM al estudio de láminas lípido-proteicas y cristales bidimensionales permite el estudio estructural de proteínas de membrana, así como la posibilidad de estudiar en tiempo real y en condiciones biomiméticas este tipo de moléculas, permitiendo, en algunos casos, el diseño de biosensores.La hipótesis de trabajo y objetivo principal de esta Tesis fue el estudio detallado del proceso de reconstitución de dos proteínas transmembranarias, la porina Omp1 de Serratia marcescens y la lactosa permeasa de Escherichia coli (LacY), tomando como modelo para comprobar la posible aplicación de las técnicas utilizadas a este tipo de moléculas, el citocromo c (Cyt c) y la melitina (MLT).Las principales conclusiones a las que se llegó en este trabajo fueron:· Las proteínas Omp1 y LacY presentaron una estructura y estabilidad en solución condicionadas por la presencia de surfactante adecuado.· La solubilización de liposomas con los surfactantes OG y DDM fue estudiada con las composiciones lipídicas: POPC, DMPC:POPC (1:1, mol:mol), POPE:POPG (3:1, mol:mol) y extracto lipídico de E. coli, en las que incorporaron las proteínas de membrana, permitiendo obtener tanto proteoliposomas como láminas lípido-proteicas.· La incorporación de Cyt c, MLT, Omp1 y LacY a la bicapa lipídica produce, en los casos estudiados, variaciones positivas del valor del potencial electrostático de superficie.· Los valores de "delta-phi" demuestran que la incorporación de las proteínas estudiadas se produce tanto por interacción electrostática, como por su balance hidrofília/lipofília.· Se observa una rigidificación de la bicapa lipídica al incorporar la LacY, debida a la reorganización de los lípidos, para conferir estabilidad a la proteína.· La incorporación de la LacY puede inducir la segregación lateral de fosfolípidos, al formarse regiones enriquecidas en las mismas.· La proteína LacY, en forma de dímero, fue cristalizada en 2D, obteniéndose los parámetros de celda: a = 13,15 nm, b = 16,74 nm, gamma = 116º.· La incorporación óptima de proteínas de membrana para dar lugar a cristales 2D, se produce en fosfolípidos como la POPC, que se encuentran en estado fluido durante todo el proceso de cristalización y que no presentan dominios lipídicos.· La inmovilización de proteínas sobre sustratos adecuados es un factor crítico en el control y manipulación molecular necesarios para posibles aplicaciones, como podría ser el diseño y fabricación de biosensores. / Membrane proteins constitute the 25-30 % of the total proteins cell that can be implicated in processes as drug resistance. The biomimetic models, proteoliposomes and 2D crystals are useful tools to study this kind of proteins.The knowledge about phenomena implicated in reconstitution of membrane proteins into liposomes is limited. Basically, this process is empiric, the presence of liposomes and detergents to form mixed micelles is required, and they are also needed membrane proteins with a high purity degree. These conditions are crucial to study the incorporation of membrane proteins and their insertion into the bilayer, including, in some cases its physiological activity.The application of nanometric techniques, as atomic force microscopy (AFM), and some methods of fluorimetry (obtention of increase of membrane surface potential and fluorescence anisotropy), allow the study of the reconstitution process of membrane proteins. The AFM application to proteolipid sheets and bidimensional crystals lead to obtain structural information of membrane proteins. Additionally there is the possibility to perform studies in real-time and under biomimetical conditions. This fact opens the possibility to design a biosensors based in these proteins. The hypothesis and main objective of this work was the detailed study of the reconstitution process of two transmembrane proteins, the porine Omp1 of Serratia marcescens and lactose permease of Escherichia coli (LacY). cytochrome c and the peptide melitin were used as models to assure the possible application of fluorescence techniques to these proteins.Finally, the main conclusions were:· Liposomes and proteolipid sheets were stables in solution after being extended onto a flat solid substrates. · LacY reconstituted in POPC was crystallized in 2D and visualized using AFM, obtaining a dimmer as a motif. The cell parameters were: a = 13.15 nm, b = 16.74 nm, gamma = 116º, accordingly at a p2 symmetry.

Fundamento y objetivo: El acoso laboral sistemático (ACS), a pesar de las diferentes tratamientos farmacológicos a largo plazo y tratamientos no farmacológicos, conduce a una alta tasa de incapacidades permanentes y graves trastornos mentales. El objetivo de esta comunicación es analizar la prescripción de medicamentos para el tratamiento de enfermedades, trastornos, síntomas y signos que experimentan los afectados de ACSa, teniendo en cuenta el diagnóstico establecido por los MAP o psiquiatras. Configuración y método: Estudio observacional, longitudinal, basado en la revisión de los registros médicos de las personas afectadas por HSW. Principales medidas de resultado: datos socio-demográficos, días de incapacidad temporal, el período de HSW psicológica, los principales diagnóstico de acuerdo a la Clasificación Internacional de Enfermedades (CIE-10), la prescripción patrón de acuerdo con la clasificación ATC. Resultados: 50 pacientes fueron incluidos en el estudio (edad media: 45,5 años; 62% mujeres). Los períodos medios de ACS y de ILT eran 4 años y 413 días respectivamente. Los principales diagnósticos fueron 76,5% F40-48 y el 23,1% F30-39 por médico de cabecera y el 49,4% F30-39 y del 42,1% F40-48 por el psiquiatra. El número total de prescripciones fue 909, incluyendo 67 diferentes principios activos, principalmente del grupo N de la clasificación ATC (88%), dirigidos a mitigar los síntomas mentales y lograr la remisión completa de los síntomas de la enfermedad mental. El restante (12%) se utiliza para tratar el estado psicosomático aparato locomotor, digestivo, cardiovascular, endocrino, dermatológicas infecciosas, respiratorias y los trastornos de dolor crónico. Los psiquiatras se centran su actividad en la depresión y, así, su prescripción es el 49% de los antidepresivos en comparación con el 29% de los ansiolíticos. Además, la prescripción de antidepresivos aproximadamente coincide con el porcentaje de pacientes con diagnóstico de depresión (43%), mientras que el porcentaje de casos diagnosticados de ansiedad (53%) es muy superior que receta ansiolíticos. Sin embargo, la Guía NICE para los estados depresivos que en el caso de los asociados síntomas de la depresión y la ansiedad, la depresión merece una atención prioritaria. Conclusiones: Los diagnósticos de los psiquiatras son consistentes con los criterios establecidos en la literatura internacional al describir los trastornos mentales graves y discapacitantes que afectan a HSW psicológico. La prescripción de antidepresivos está de acuerdo con el enfoque de diagnóstico informado por todos los especialistas, y también está de acuerdo con las recomendaciones de NICE Directrices para la depresión. / Objectives: The first aim of this study was to examine the possible association of long standing Systematic psychological harassment at work (SPHW) with psychiatric disorders and its characteristics. The second aim was to analyze drug prescription used to treat diseases, disorders, symptoms, and signs of those affected with psychological SPHW, taking into account the established diagnosis by the General Physician (GP) or Psychiatrists. Method and study subjects: Design: Longitudinal observational study with follow up focused on persons undergoing a long duration SPHW; based on the review of medical records of people affected by SPHW Total length of the follow up period was 6 years, from 2006 to 2012. Mean duration of follow up was 3.1 years (range 1.1-6.3 years). Setting: Patients who voluntarily attend to a Catalonia Harassment Victims association (Associació de Víctimes d’Assejament laboral de Catalunya: AVALC) asking for psychological and laboral aid. Subjects: 132 persons were recruited along a seven years period, 84 women (63.6%) and 48 men (36.4%). Mean age was 45.5 years (SD ± 9.8), 43.8 years for women and 48.3 years for men (p= 0.114). SPHW was measured by self-reported data. All participants were assessed using several neuropsychological instruments such as the Beck Scale, the Hamilton Depression Scale, the Echeburua Scale, the PDS (Posttraumatic Diagnostic Scale) and the LIPT (Leymann Inventory of Psychological Terrorization). Medical records were examined to verify all the diagnosis affecting to the participants. After checking the medical records, only 50 cases (37.3%), 31 women and 19 men, had verified psychiatric diagnostics, according to ICD-10 criteria. Main outcome measures: - Socio-demographic data: age, gender - Work environment: types of harassers, mainly distributed by work sectors, public administration workers and private organizations workers, days of temporary disability, period of SPHW. - Main diagnosis according to International Classification of Diseases ICD-10 - Type of assistance: primary care, mental health unit, emergency department, neurologist or occupational health unit. - Previous pharmacotherapeutical history including all prescribed drugs (generic or brand) related with working problems and concomitant medication, number of prescriptions, active principles (DCI), reason for prescription, dose and dosing interval, duration of treatment, and number of drugs per patient. - Established treatment planning (pharmaceutical care) designed to detect, prevent and solve potential or acute problems related with the prescribed drugs, such as adherence, interactions, collateral effects, negative outcomes, effectiveness and safety. - Prescription pattern according to Anatomical, Therapeutic, Chemical Classification System (ATC classification). - Most prescribed active principles and percentages of utilization. Key terms: Systematic psychological harassment at work, mobbing, workplace bullying, stress, occupational health, work environment, psychosocial factors mental health disorders, gender, posttraumatic stress disorder, major depression disorder, depression, anxiety, comorbidity, insomnia, somatic disorder, prevalence, epidemiology, cost mental disorder, treatment / pharmacotherapy/ drug, compliance. Background/Introduction The phenomenon of systematic psychological harassment at work (SPHW) first described by Heinz Leymann, refers to systematic and prolonged exposure to frequent hostile behaviours at work. SPHW is commonly considered as an expression of violent behaviour. SPHW has been defined as ‘‘harassing, offending, socially excluding someone or negatively affecting someone’s work tasks...” it has to occur repeatedly and regularly (e.g., weekly) and over a period of time (e.g., about six months). However, there is no accepted common definition as yet and different terminology is used as synonym of bullying (e.g., harassment, mobbing, emotional abuse, victimisation, psychological abuse at work). Research about this topic has dramatically increased in the last years. These behaviours may lead to the stigmatisation and victimization of the exposed individuals. A reciprocal relationship between SPHW and mental distress has been described. Several studies have shown that SPHW is associated with mental distress. Furthermore, mental distress was shown to be a predictor of bullying. It seems that experiencing workplace bullying can contribute to mental distress, but individual characteristics may also contribute to the perception of bullying. Thus, a significant relationship between general proneness to negative feelings and work victimization has been described. A theoretical framework, the cognitive theory of trauma, proposes the post-traumatic stress disorder (PTSD) as the result of systematic psychological harmful events. The traumatic potential of bullying is unquestionable. So, the path from job demands to PTSD through bullying seems plausible. PTSD is a severe anxiety disorder that can be developed after exposure to any event that results in psychological trauma. Diagnostic symptoms for PTSD include re-experiencing the original trauma(s) through flashbacks or nightmares, avoidance of stimuli associated with the trauma, and increased arousal, such as difficulty falling or staying asleep, anger, and hyper vigilance. Diagnostic criteria require that symptoms last more than one month and cause significant impairment in social, occupational, or other important areas of functioning. Exposure to intense bullying at work may change the individual’s perceptions of their work-environment and life in general to one of threat, danger, insecurity and self-questioning, which may result in emotional, psychosomatic and psychiatric problems, even affecting the personal and familiar relationships of the victims. Most studies examining the relationship between psychosocial work characteristics and mental health outcomes, ranging from symptoms and psychological distress to diagnosed psychiatric disorders, have been cross-sectional and, therefore, the causal association between work stressors and mental health remains uncertain. The aim of this study is to describe and analyze the possible relationship between SPHW characteristics and the subsequent psychiatric diagnoses, such as the presence of PTSD symptoms as a possible consequence of workplace bullying-related victimization. PTSD in the workplace may be a common problem and there has been scarce research about this topic (32). RESULTS Mean duration of SPHW was 4.0 years (SD ± 2.8), 3.7 years for women and 4.6 years for men (p= 0.226). 14% of participants suffered SPHW along 3 years or less, 48% from 3 to 6 years, 30% from 6 to 9 years and 8% more than 9 years. In 62.7% of cases, the harassers were hierarchic superiors and in 9.4% of cases companions and/or collaborators contributed to bullying. In 62.7% of cases, the harassers were hierarchic superiors and in 9.4% of cases companions and/or collaborators contributed to bullying. Cases were mainly distributed by work sectors as follows: 28.4% public administrations, 23.2% commercial sector, 23.2% industrial sector and 14.0% services. In our sample, we included 35.6% of public administration workers and 64.4% of private organizations workers. Despite of the greater proportion of private organization workers, we found a high number of worked years in the same organization: 25% of cases with 20 or more years worked at the same organization. The distribution was as follows: 8.3% less than 1 year, 25.0% from 1 to 5 years, 26.5% from 5 to 10 years, 6.8% from 10 to 15 years, 8.3% from 15 to 20 years, 12.1% from 20 to 25 years, 4.5% from 25 to 30 years, 6.1% from 30 to 35 years and 2.3% from 35 to 40 years. The mean number of diagnoses for each patient was 4.2 (range 1-7). Persons with previous mental disorders were excluded. The remaining participants met no criteria or were lost along the follow-up period. All of them showed a follow-up period longer than 30 months. Mean duration of follow up for this group was 3.8 years (range 2.7-6.3 years). The main diagnoses were 76.5% F40-48 and 23.1% F30-39 by GP, and 49.4% F30-39 and 42.1% F40-48 by psychiatrist. The most frequent psychiatric diagnoses were Posttraumatic Stress Disorder (PTSD) was the most often diagnosed disorder (44.0%). Major depression has been developed by 20% of patients with important comorbidity, mainly physical symptoms as insomnia, headache, digestive disorders, etc. Significant differences were found in the association of SPHW length with type of subsequent psychiatric diagnosis (p=0.015) and public/private job (p=0.047). Psychiatric diagnoses by gender showed significant differences for PTSD (p=0.043), generalized anxiety (p=0.041) and minor depression episodes (p=0.049). One case of suicide was registered (0.8%) and 3 cases of autolysis attempts (2.3%). All participant patients presented sick leaves. Mean length of temporary disability was 13.8 months (SD ±6.7), 13.2 months for women and 14.8 months for men. Sick leaves duration was slightly higher for men, but no significant differences by gender were observed (p= 0.417). On the other hand, significant differences were observed in the mean duration of sick leaves by diagnosis. Mean values were 9.5 months for adaptation disorders, 11.6 months for generalized anxiety, 11.7 months for minor depression episodes, 11.9 months for major depression and 17.2 months for PTSD (p=0.019). We also found significant differences in the mean duration of sick leaves by public/private job. Mean values were 11.7 months for private jobs and 17.6 months for public employments (p=0.002). Significant differences were found in the mean age of the psychiatric diagnoses. Cases with diagnosis of adaptation disorders were the youngest and with diagnosis of major depression were the oldest (p=0.019). SPHW length seems related with the type of subsequent psychiatric diagnosis. The mean SPHW length was longer for the more severe disorders, PTSD and depression, and shorter for anxiety and adaptation disorders. So, 1.9 years for adaptation disorders, 2.2 years for generalized anxiety, 3.9 years for minor depression, 4.6 years for major depression and 5.1 years for PTSD (p=0.015). Significant differences were also found in the association between SPHW length and public/private job. Mean values were 3.4 years for private jobs and 5.0 years for public jobs (p=0.047). There were 909 prescriptions, including 67 different active principles (88%). Most of them were from the N group of the ATC classification, which targeted a mitigation of mental symptoms and achievement of complete remission. The remaining 12% prescriptions were used to treat psychosomatic symptoms including locomotive, digestive, cardiovascular, endocrine, infectious, dermatological, respiratory, and chronic pain disorders. In our study, 92% of patients are treated with both anxiolytic and antidepressant drugs. General Physicians focus on anxiety, these accounts for 70% of anxiolytic prescribing, in comparison to them being responsible for 20% of antidepressant prescribing. Nevertheless, the NICE Guidelines for Depression state that if a case is presented in which symptoms of both depression and anxiety coexist, the former deserves priority attention. Psychiatrists focus on depression; this accounts for 49% of antidepressant prescribing, in comparison to them being responsible for 26.3% of anxiolytic prescribing. Furthermore, the number of antidepressants prescribed is roughly in accordance with the percentage of patients diagnosed with depression (43%), while the percentage of diagnosed anxiety cases (53%) is higher than the amount of anxiolytics prescribed. Treatment pattern has been successful in 34% of SPHW with comorbidity, post-traumatic stress syndrome (PTSD) and major depression episodes (MDE). Gender differences have been observed in the successful rates: 47.4% for men and 25.8% for women. 38% of SPHW with associated major depression disorder (MDD) have achieved a remission. Gender differences have been observed in the successful rates: 47.4% for men and 32.3% for women. The total number of patients in not remission is 17 (34%), 10 women (20%) and 7 men (36.8%). These patients are treated with both antipsychotic drug (N05A) and another antidepressant (N06AX) drugs. Antipsychotic drugs are almost exclusively prescribed by psychiatrists in 88%. DISCUSSION An important limitation of the study is its lack of generalization, because we have focused on diagnosed cases and not in general population. But, in the other hand, there is scarce research focused on persons with a long history of intense bullying. A second limitation is that the data were self-reported, which raises the issue of common method variance. However, other methods, such as observer ratings of working conditions, may be equally affected by bias. Participants in the study have undergone very long periods of SPHW, so it can be expected that psychological or psychiatric consequences should be more severe than those reported by other studies. It has been described a significant correlation between the duration and intensity of SHPW and the subsequent psychiatric symptoms. PTSD is a severe consequence of important traumatic events. Heterogeneity in PTSD diagnosis is a complex factor which should be assessed. It is not the purpose of this paper to solve the complex issue of the appropriateness of PTSD diagnosis as a consequence of bullying, which is related to the conceptualization of bullying as an overwhelming traumatic event. However, association between SPHW and PTSD have been reported by a large number of studies, most of them having a cross-sectional design. In our data, PTSD incidence in patients SPHW is higher than the described in the literature. It may be partially due to the longitudinal design of the study. The long length of SPHW jointly with the prolonged follow up period (mean value: 3.1 years) could lead into a higher PTSD development ratio. A key question in this relationship is how long do the psychological effects of bullying persist over time, independently of SPHW duration. This question remains uncertain and reveals the need of longitudinal studies with long enough periods of follow up. Association between depression and SPHW is well documented. Moreover, a meta-analysis collecting epidemiological data calculated summary estimates for the effects of work stress: the average risk of depression across all studies weighted by the number of participants was 1.2 to 1.4 fold. Our results show a 20% incidence of major depression cases, probably due to the characteristics of SPHW, specially its long length. Gender has been found to be the strongest predictor of Post-Stress Traumatic Disorder (PSTD). Our results agree with the important role of gender. Although WPB duration seems to be highly correlated with the development of PTSD, our data show longer length of SPHW with lower incidence of PTSD for males and shorter length of SPHW with higher rates of PTSD for females. A great number of patients were lost along the follow-up period (32.1%) and, so, it could not be verified the correct diagnosis, if any, and its possible relation with SPHW. However, the heavy utilization of psychotropic drugs seems to be a strong indicator of severe mental disorders. The high number of withdrawals may be probably due to the characteristics of participants. Persons attending a victims association in a desperate situation, after a long suffering time, need strong and urgent aid. Solving the SPHW problem is a complex and difficult question, so patients go on looking for new options. Comorbidity is the main prognosis factor for transitory disability length. The high number of diagnoses for each patient observed in our study probably influences the long length of sick leaves. Major psychiatric disorders are commonly related to work disability and, usually, they are an important potential factor for unemployment. It has been argued that the association between work characteristics and health is due to confounding by social class. Thus, poor work environmental characteristics are merely a marker of low socioeconomic position. But even after adjustment for social position, in occupational cohorts of homogeneous social status, work characteristics are still important determinants of psychological distress. Mental health problems can take years to develop but the cross-sectional studies explore a reduced time period. In our data the minimum follow-up period is 30 months, so the described relationship between WPB and psychiatric disorders should be more consistent. CONCLUSIONS Longer length of SPHW is associated with more severe psychiatric diagnoses. As mental health problems can take years to develop, long follow up periods are needed to identify the psychiatric consequences of SPHW. The diagnoses made by psychiatrists were consistent, but GP diagnoses weren’t it, with the criteria established in the international literature describing severe and disabling mental disorders affecting psychological of SPHW. The prescribing of antidepressants correlated with the diagnostic approach used by specialists, and was also in accordance with the NICE Guidelines for depression. SPHW despite of different long-term pharmacological and non-pharmacological treatments, leads to a high rate of permanent disabilities and severe mental disorders. Prescription of drugs in Mental Health settings by GP is based on so-called target symptoms (depression, anxiety, insomnia and somatic) representing 93.9% of total prescription. However, Psychiatrists prescription is based on an another target symptoms (psychosis diseases, depression, anxiety, insomnia, suicide risk, aggressive behaviour, attention, phobia, mania, neurosis, agitation, comorbid psychiatric disorders) Chronic insomnia in patients with major depression disorder (MDD) is underdiagnosed and undertreated, despite of its high prevalence in this type of patients with SPHW.