Anales de la RANM

40 A N A L E S R A N M R E V I S T A F U N D A D A E N 1 8 7 9 S U P L E M E N T O I SIMPOSIO · JÓVENES INVESTIGADORES Libro de Abstracts An RANM. 2021;138(03).supl01: 40 - 54 en modelos relacionados con el estrés oxidativo. Además, llevamos a cabo un modelo de rodajas de estriado de rata adulta con rotenona y 6-hidroxido- pamina como tóxicos. Los compuestos restauraron la muerte celular, estrés oxidativo y modularon proteínas relacionadas con el proceso neurodegener- ativo. Fueron capaces también de revertir el proceso inflamatorio inducido por LPS en cultivos primarios de glía, disminuyendo la producción de óxido nítrico y citocinas pro-inflamatorias. Nuestro estudio también incluyó un programa in silico para caracterizar los mecanismos de inhibición de MAO-B. A partir de esta información, el objetivo es tratar de mejorar la potencia de inhibición de MAO-B, reducir la toxicidad celular y mejorar las propie- dades farmacológicas en cuanto a biodisponibilidad y permeabilidad de la barrera hematoencefálica de los nuevos compuestos. Para ello, hemos construido una nueva quimioteca de unos 500 compuestos para llevar a cabo un proceso de cribado virtual. Tras la evaluación de los resultados, hemos selecci- onado una nueva familia de 30 compuestos, que han sido sintetizados y sometidos a una evaluación farmacológica completa. Estos nuevos compuestos mostraron un mejor perfil con un aumento en la potencia de inducción de NRF2, selectividad de MAO-B y permeabilidad al sistema nervioso central por difusión pasiva. También demostraron un mejor perfil neuroprotector, una toxicidad celular reducida y mantuvieron propiedades antioxidantes y antiinflama- torias complementarias. Además de esto, hemos diluci- dado los mecanismos para controlar la homeostasis proteica del compuesto líder de esta segunda familia, mostrando una activación de la vía del sistema ubiqui- tina-proteasoma y la macroautofagia. Parkinson's disease (PD) is the second most prevalent neurodegenerative disease. The lack of effective treatments able to stop the progression of the disease highlights the importance of finding new therapies. Physiopathological features such as dopamine depletion and selective destruction of dopaminergic neurons, together with the presence of α-synuclein protein deposits, are considered main events in PD. Importantly, oxidative stress plays a major role in the disease progression leading to deregulation of protein processing, mitochondrial damage, and neuroinflam- mation. These factors, in turn, lead to the increase of oxidative stress generating a loop that accelerates the neurodegeneration process. The broad spectrum of physiological roles regulated by transcription factor NF-E2-related factor 2 (NRF2) positions it as a good target for the develo- pment of innovative treatments for chronic diseases characterized by a complex network of pathological pathways. In that sense, we have developed two families of new NRF2 inducers with complemmen- tary activities for the treatment of PD, regarding that NRF2 regulates a plethora of cytoprotective genes related to oxidative stress, inflammation, and protein homeostasis, among other processes involved in the onset of PD. Apart from this, the novel compounds are also designed to selectively inhibit monoamine oxidase B (MAO-B). This enzyme is involved in controlling oxidative stress, regarding dopamine metabolism, and MAO-B inhibitors have been used for years in clinic to treat PD symptoms. Based on this, we have a first small chemical library of 9 new multitarget compounds combining several activities: NRF2 inducing capacity, MAO-B potent and selective inhibition, and neuroprotective ability in oxidative stress-related models. In addition, we performed a more complex model of adult rat striatal slices treated with rotenone and 6-hydroxydopa- mine as toxics. Selected compounds were protective in terms of cell death, oxidative stress, and modula- tion of several proteins related with neurodegenera- tion. Hit compounds were also capable of restoring LPS-induced inflammatory conditions in primary glial cultures leading to a decrease in the production of nitric oxide and pro-inflammatory cytokines. Our study also included an in silico program to charac- terize the mechanisms of MAO-B inhibition. Regarding the information from this first family of compounds, the next objective is mainly trying to improve the potency for MAO-B inhibition, reduce cell toxicity, and also improve drug-like proper- ties regarding half-life and blood-brain barrier permeability of the novel compounds. To this end, we have built a new chemical library of almost 500 compounds and subjected it to a virtual screening program. After evaluation of the results, we selected a new family of 30 compounds, we synthesized them, and we performed a complete pharmacolo- gical evaluation. These novel compounds showed a better profile with improved NRF2 induction potency, MAO-B selectivity, and central nervous system permeability by passive diffusion. They also demonstrated a better neuroprotective profile, reduced cell toxicity, and maintained complementary antioxidant and anti-inflammatory-related proper- ties. In addition to this, we have also elucidated the mechanisms for controlling protein homeostasis of the lead compound from this family, showing an upregulation of the ubiquitin-proteasome system pathway and activation of macroautophagy. DESARROLLO DE NUEVOS COMPUESTOS MUL- TIDI ANA DIRIGIDOS AL TRATAMIENTO DE LA ENFERMEDAD DE ALZHEIMER DEVELOPMENT OF NEW MULT I TARGET COMPOUNDS FOR THE TREATMENT OF ALZHE IMER´S D I SEASE Enrique Crisman 1,2,3 , Sheila Abril 1 , Laura Vozmediano 1 , Esteban Daudén 1 and Rafael León 1 1 Instituto de Química Médica, Consejo Superior de Investiga- ciones Científicas (IQM-CSIC). 2 Instituto Teófilo Hernando y Departamento de Farma- cología y Terapéutica, Facultad de Medicina. Universidad Au- tónoma de Madrid. 3 Instituto de Investigación Sanitaria La Princesa (IIS-IS), Hos- pital Universitario de la Princesa.