Loss of mitochondrial Ndufs4 in striatal medium spiny neurons mediates progressive motor impairment in a mouse model of leigh syndrome

Altres ajuts: Juan del la Cierva (IJCI-2015-24576)Inability of mitochondria to generate energy leads to severe and often fatal myoencephalopathies. Among these, Leigh syndrome (LS) is one of the most common childhood mitochondrial diseases; it is characterized by hypotonia, failure to thrive, respiratory insufficiency and progressive mental and motor dysfunction, leading to early death. Basal ganglia nuclei, including the striatum, are affected in LS patients. However, neither the identity of the affected cell types in the striatum nor their contribution to the disease has been established. Here, we used a mouse model of LS lacking Ndufs4, a mitochondrial complex I subunit, to confirm that loss of complex I, but not complex II, alters respiration in the striatum. To assess the role of striatal dysfunction in the pathology, we selectively inactivated Ndufs4 in the striatal medium spiny neurons (MSNs), which account for over 95% of striatal neurons. Our results show that lack of Ndufs4 in MSNs causes a non-fatal progressive motor impairment without affecting the cognitive function of mice. Furthermore, no inflammatory responses or neuronal loss were observed up to 6 months of age. Hence, complex I deficiency in MSNs contributes to the motor deficits observed in LS, but not to the neural degeneration, suggesting that other neuronal populations drive the plethora of clinical signs in LS

Defined neuronal populations drive fatal phenotype in a mouse model of leigh syndrome

Altres ajuts: Seattle Children's Research Institute: Seed Funds;NINDS: R01 NIH/NS 102796; University of Washington Neurological Surgery Department: Ellenbogen Neurological Surgery Research Funds; University of Washington: The Ryan J. Murphy SUDEP Research Funds; Mitochondrial Research Guild: Seed FundsMitochondrial deficits in energy production cause untreatable and fatal pathologies known as mitochondrial disease (MD). Central nervous system affectation is critical in Leigh Syndrome (LS), a common MD presentation, leading to motor and respiratory deficits, seizures and premature death. However, only specific neuronal populations are affected. Furthermore, their molecular identity and their contribution to the disease remains unknown. Here, using a mouse model of LS lacking the mitochondrial complex I subunit Ndufs4, we dissect the critical role of genetically-defined neuronal populations in LS progression. Ndufs4 inactivation in Vglut2expressing glutamatergic neurons leads to decreased neuronal firing, brainstem inflammation, motor and respiratory deficits, and early death. In contrast, Ndufs4 deletion in GABAergic neurons causes basal ganglia inflammation without motor or respiratory involvement, but accompanied by hypothermia and severe epileptic seizures preceding death. These results provide novel insight in the cell type-specific contribution to the pathology, dissecting the underlying cellular mechanisms of MD

Multi-Target Directed Donepezil-Like Ligands for Alzheimer's Disease

HIGHLIGHTS Alzheimer's disease (AD), the most common form of adult onset dementia, is an age-related neurodegenerative disorder characterized by progressive memory loss, decline in language skills, and other cognitive impairments. Although its etiology is not completely known, several factors including deficits of acetylcholine, β-amyloid deposits, τ-protein phosphorylation, oxidative stress, and neuroinflammation are considered to play significant roles in the pathophysiology of this disease. For a long time, AD patients have been treated with acetylcholinesterase inhibitors such as donepezil (Aricept®) but with limited therapeutic success. This might be due to the complex multifactorial nature of AD, a fact that has prompted the design of new Multi-Target-Directed Ligands (MTDL) based on the "one molecule, multiple targets" paradigm. Thus, in this context, different series of novel multifunctional molecules with antioxidant, anti-amyloid, anti-inflammatory, and metal-chelating properties able to interact with multiple enzymes of therapeutic interest in AD pathology including acetylcholinesterase, butyrylcholinesterase, and monoamine oxidases A and B have been designed and assessed biologically. This review describes the multiple targets, the design rationale and an in-house MTDL library, bearing the N -benzylpiperidine motif present in donepezil, linked to different heterocyclic ring systems (indole, pyridine, or 8-hydroxyquinoline) with special emphasis on compound ASS234, an N -propargylindole derivative. The description of the in vitro biological properties of the compounds and discussion of the corresponding structure-activity-relationships allows us to highlight new issues for the identification of more efficient MTDL for use in AD therapy

Effectiveness of an intervention for improving drug prescription in primary care patients with multimorbidity and polypharmacy:Study protocol of a cluster randomized clinical trial (Multi-PAP project)

This study was funded by the Fondo de Investigaciones Sanitarias ISCIII (Grant Numbers PI15/00276, PI15/00572, PI15/00996), REDISSEC (Project Numbers RD12/0001/0012, RD16/0001/0005), and the European Regional Development Fund ("A way to build Europe").Background: Multimorbidity is associated with negative effects both on people's health and on healthcare systems. A key problem linked to multimorbidity is polypharmacy, which in turn is associated with increased risk of partly preventable adverse effects, including mortality. The Ariadne principles describe a model of care based on a thorough assessment of diseases, treatments (and potential interactions), clinical status, context and preferences of patients with multimorbidity, with the aim of prioritizing and sharing realistic treatment goals that guide an individualized management. The aim of this study is to evaluate the effectiveness of a complex intervention that implements the Ariadne principles in a population of young-old patients with multimorbidity and polypharmacy. The intervention seeks to improve the appropriateness of prescribing in primary care (PC), as measured by the medication appropriateness index (MAI) score at 6 and 12months, as compared with usual care. Methods/Design: Design:pragmatic cluster randomized clinical trial. Unit of randomization: family physician (FP). Unit of analysis: patient. Scope: PC health centres in three autonomous communities: Aragon, Madrid, and Andalusia (Spain). Population: patients aged 65-74years with multimorbidity (≥3 chronic diseases) and polypharmacy (≥5 drugs prescribed in ≥3months). Sample size: n=400 (200 per study arm). Intervention: complex intervention based on the implementation of the Ariadne principles with two components: (1) FP training and (2) FP-patient interview. Outcomes: MAI score, health services use, quality of life (Euroqol 5D-5L), pharmacotherapy and adherence to treatment (Morisky-Green, Haynes-Sackett), and clinical and socio-demographic variables. Statistical analysis: primary outcome is the difference in MAI score between T0 and T1 and corresponding 95% confidence interval. Adjustment for confounding factors will be performed by multilevel analysis. All analyses will be carried out in accordance with the intention-to-treat principle. Discussion: It is essential to provide evidence concerning interventions on PC patients with polypharmacy and multimorbidity, conducted in the context of routine clinical practice, and involving young-old patients with significant potential for preventing negative health outcomes. Trial registration: Clinicaltrials.gov, NCT02866799Publisher PDFPeer reviewe

Study of new propargylamine and donepezil-derived compounds as multitarget agents for the treatment of alzheimer’s disease

El compuesto PF9601N es un derivado de propargilamina y un potente inhibidor irreversible de la enzima monoamino oxidasa B (IMAO-B) el cual fue identificado por nuestro grupo tras una extensiva búsqueda de potenciales IMAOs. Además de su potente capacidad inhibidora, el PF9601N posee varias propiedades neuroprotectoras demostradas en varios modelos animales y celulares de la enfermedad de Parkinson (EP). Estos efectos, los cuales han sido relacionados con la presencia de la propargilamina en su estructura, están mediados por acciones en vías involucradas en la neurodegeneración observada en otras enfermedades neurodegenerativas como la enfermedad de Alzheimer (EA). Así, para estudiar más en detalle las propiedades beneficiosas del PF9601N investigamos sus efectos en un modelo in vivo de excitotoxicidad, un mecanismo implicado en el daño neuronal observado en las enfermedades neurodegenerativas. El hallazgo de que el PF9601N era capaz de evitar el daño excitotóxico mediante la disminución de la liberación inducida de glutamato y aspartato, y el aumento de la liberación de taurina así como mediante la prevención de la activación glial y la apoptosis proporcionó un valor añadido a este compuesto para ser considerado en la terapia de estas enfermedades. El tratamiento actual para la EA se basa principalmente en el uso de inhibidores de las enzimas colinesterasas (IChEs). Sin embargo, estos fármacos no son capaces de disminuir la progresión de la enfermedad y sólo producen una mejora temporal de los síntomas. Actualmente está ampliamente aceptado que la EA es una enfermedad multifactorial. En este contexto, la aproximación farmacológica más novedosa, conocida como aproximación de los MTDL (de las siglas en inglés “ligandos dirigidos hacia múltiples dianas”), propone el uso de compuestos multifuncionales capaces de abrazar varias propiedades biológicas. Esta tesis se centra en el estudio de la relación estructura-actividad (REA) así como la evaluación biológica de varios compuestos híbridos especialmente diseñados y sintetizados para actuar sobre múltiples factores involucrados en la EA. Los compuestos híbridos combinan la porción de bencilpiperidina de Donepecilo, un anticolinesterásico ampliamente utilizado en el tratamiento de la enfermedad, con el grupo propargilamina o indolil propargilamina presente en PF9601N, con el objetivo de obtener un compuesto capaz de retener la capacidad inhibidora de MAO así como las propiedades neuroprotectoras y antiapoptóticas de PF9601N. El trabajo presentado en esta tesis demuestra que algunos de los compuestos híbridos son potentes IMAOs (rango nM) y moderadamente potentes IChEs (rango subM). De entre todos los compuestos evaluados, ASS234 resultó ser un potente inhibidor de la agregación del péptido β−amiloide (A) y fue capaz de ejercer una acción protectora frente a la toxicidad inducida por Aβ y H2O2 en células neuronales. En resumen, los datos presentados en esta tesis doctoral sugieren que el compuesto ASS234 es un compuesto multidiana muy prometedor que podría tener un papel modificador en la EA dada su demostrada capacidad de interactuar con varias dianas involucradas en la patogénesis de esta enfermedad.PF9601N is a propargylamine-containing irreversible monoamine oxidase B inhibitor (MAOBI) previously identified by our group in an extensive screen of potential MAOIs. Besides its potent inhibitory capacity, it possesses several neuroprotective properties demonstrated in different animal and cellular models of Parkinson’s disease (PD). The beneficial effects of PF9601N, which have been related to the propargylamine group present in the molecule, are mediated through actions in pathways that are commonly involved in the neurodegeneration observed in other neurodegenerative disorders such as Alzheimer’s disease (AD), thus making this molecule a promising agent in the therapy of this disease as well. Thus, to study the beneficial properties of PF9601N in depth, we investigated its effects against an in vivo model of excitotoxicity, an important mechanism involved in the neuronal damage observed in neurodegenerative diseases. The finding that PF9601N was able to prevent the induced excitotoxic damage by decreasing the evoked release of excitatory neurotransmitters and decreasing the output of the inhibitory and neuroprotective taurine as well as preventing the induced glial activation and apoptosis gave more value to this compound to be considered in the therapy. The current treatment for AD is the use of cholinesterase inhibitors (ChEIs) although there is also a NMDA receptor antagonist. However, far from stopping the disease’s progression, these drugs only produce a temporary symptomatic benefit, thus highlighting an urgent need to provide real disease-modifying drugs. At present, the most accepted notion is that AD is a multifactorial disease caused by many different factors and thus drug therapy with multifunctional compounds, the so-called multi-target-directed ligand (MTDL) approach, embracing diverse biological properties will have noticeable advantages over individual-target drugs or cocktails of drugs. In this context, this thesis focuses on the structure-activity relationship (SAR) study and the biological evaluation of different hybrid compounds specifically designed and synthesised to target multiple factors involved in AD. The hybrid molecules combine the benzyl piperidine moiety of Donepezil, a commonly used anticholinesterasic for the treatment of AD, with the propargylamine or the indolyl propargylamine substructure of PF9601N, with the aim of retaining the MAO inhibitory capacity as well as the neuroprotective and antiapoptotic properties observed for this compound. The work presented in this thesis demonstrates that some hybrid compounds are potent MAOIs (nM range) and moderately potent ChEIs (submicroM range). Among them, ASS234 has also been shown to reduce Αβ fibrillogenesis, and to protect neuronal cells from A and H2O2 toxicity. Thus, this compound has proved to be able to block the Aβ-induced cell death in two ways: by preventing caspase cleavage and activation and blocking LDH release. Overall, the present data suggest ASS234 as a promising MTDL that may have a potential disease-modifying role in the treatment of AD since it is able to interact with diverse targets involved in the pathogenesis underlying AD

Nuevos derivados de propargilamina con capacidad neuroprotectora para el tratamiento de las enfermedades de Alzheimer y Parkinson

La presente invención se refiere a una serie derivados de N-metil-N-{[(1 -metil-5-alcoxi) -1 H-indol-2-il]metil}prop-2- ino-1-amina, que son inhibidores multipotentes de las enzimas monoaminooxidasas A y B, acetilcolinesterasa y butirilcolinesterasa, con posible aplicación dentro del ámbito de la industria farmacéutica, como fármacos para curar, detener o paliar enfermedades neurodegenerativas, tales como las enfermedades de Alzheimer y Parkinson.Peer reviewedConsejo Superior de Investigaciones Científicas (España), Universidad Autónoma de Barcelona, Universidad de BarcelonaA1 Solicitud de patente con informe sobre el estado de la técnic

Study of new propargylamine and donepezil-derived compounds as multitarget agents for the treatment of alzheimer's disease

El compuesto PF9601N es un derivado de propargilamina y un potente inhibidor irreversible de la enzima monoamino oxidasa B (IMAO-B) el cual fue identificado por nuestro grupo tras una extensiva búsqueda de potenciales IMAOs. Además de su potente capacidad inhibidora, el PF9601N posee varias propiedades neuroprotectoras demostradas en varios modelos animales y celulares de la enfermedad de Parkinson (EP). Estos efectos, los cuales han sido relacionados con la presencia de la propargilamina en su estructura, están mediados por acciones en vías involucradas en la neurodegeneración observada en otras enfermedades neurodegenerativas como la enfermedad de Alzheimer (EA). Así, para estudiar más en detalle las propiedades beneficiosas del PF9601N investigamos sus efectos en un modelo in vivo de excitotoxicidad, un mecanismo implicado en el daño neuronal observado en las enfermedades neurodegenerativas. El hallazgo de que el PF9601N era capaz de evitar el daño excitotóxico mediante la disminución de la liberación inducida de glutamato y aspartato, y el aumento de la liberación de taurina así como mediante la prevención de la activación glial y la apoptosis proporcionó un valor añadido a este compuesto para ser considerado en la terapia de estas enfermedades. El tratamiento actual para la EA se basa principalmente en el uso de inhibidores de las enzimas colinesterasas (IChEs). Sin embargo, estos fármacos no son capaces de disminuir la progresión de la enfermedad y sólo producen una mejora temporal de los síntomas. Actualmente está ampliamente aceptado que la EA es una enfermedad multifactorial. En este contexto, la aproximación farmacológica más novedosa, conocida como aproximación de los MTDL (de las siglas en inglés "ligandos dirigidos hacia múltiples dianas"), propone el uso de compuestos multifuncionales capaces de abrazar varias propiedades biológicas. Esta tesis se centra en el estudio de la relación estructura-actividad (REA) así como la evaluación biológica de varios compuestos híbridos especialmente diseñados y sintetizados para actuar sobre múltiples factores involucrados en la EA. Los compuestos híbridos combinan la porción de bencilpiperidina de Donepecilo, un anticolinesterásico ampliamente utilizado en el tratamiento de la enfermedad, con el grupo propargilamina o indolil propargilamina presente en PF9601N, con el objetivo de obtener un compuesto capaz de retener la capacidad inhibidora de MAO así como las propiedades neuroprotectoras y antiapoptóticas de PF9601N. El trabajo presentado en esta tesis demuestra que algunos de los compuestos híbridos son potentes IMAOs (rango nM) y moderadamente potentes IChEs (rango subM). De entre todos los compuestos evaluados, ASS234 resultó ser un potente inhibidor de la agregación del péptido β−amiloide (A) y fue capaz de ejercer una acción protectora frente a la toxicidad inducida por Aβ y H2O2 en células neuronales. En resumen, los datos presentados en esta tesis doctoral sugieren que el compuesto ASS234 es un compuesto multidiana muy prometedor que podría tener un papel modificador en la EA dada su demostrada capacidad de interactuar con varias dianas involucradas en la patogénesis de esta enfermedad.PF9601N is a propargylamine-containing irreversible monoamine oxidase B inhibitor (MAOBI) previously identified by our group in an extensive screen of potential MAOIs. Besides its potent inhibitory capacity, it possesses several neuroprotective properties demonstrated in different animal and cellular models of Parkinson's disease (PD). The beneficial effects of PF9601N, which have been related to the propargylamine group present in the molecule, are mediated through actions in pathways that are commonly involved in the neurodegeneration observed in other neurodegenerative disorders such as Alzheimer's disease (AD), thus making this molecule a promising agent in the therapy of this disease as well. Thus, to study the beneficial properties of PF9601N in depth, we investigated its effects against an in vivo model of excitotoxicity, an important mechanism involved in the neuronal damage observed in neurodegenerative diseases. The finding that PF9601N was able to prevent the induced excitotoxic damage by decreasing the evoked release of excitatory neurotransmitters and decreasing the output of the inhibitory and neuroprotective taurine as well as preventing the induced glial activation and apoptosis gave more value to this compound to be considered in the therapy. The current treatment for AD is the use of cholinesterase inhibitors (ChEIs) although there is also a NMDA receptor antagonist. However, far from stopping the disease's progression, these drugs only produce a temporary symptomatic benefit, thus highlighting an urgent need to provide real disease-modifying drugs. At present, the most accepted notion is that AD is a multifactorial disease caused by many different factors and thus drug therapy with multifunctional compounds, the so-called multi-target-directed ligand (MTDL) approach, embracing diverse biological properties will have noticeable advantages over individual-target drugs or cocktails of drugs. In this context, this thesis focuses on the structure-activity relationship (SAR) study and the biological evaluation of different hybrid compounds specifically designed and synthesised to target multiple factors involved in AD. The hybrid molecules combine the benzyl piperidine moiety of Donepezil, a commonly used anticholinesterasic for the treatment of AD, with the propargylamine or the indolyl propargylamine substructure of PF9601N, with the aim of retaining the MAO inhibitory capacity as well as the neuroprotective and antiapoptotic properties observed for this compound. The work presented in this thesis demonstrates that some hybrid compounds are potent MAOIs (nM range) and moderately potent ChEIs (submicroM range). Among them, ASS234 has also been shown to reduce Αβ fibrillogenesis, and to protect neuronal cells from A and H2O2 toxicity. Thus, this compound has proved to be able to block the Aβ-induced cell death in two ways: by preventing caspase cleavage and activation and blocking LDH release. Overall, the present data suggest ASS234 as a promising MTDL that may have a potential disease-modifying role in the treatment of AD since it is able to interact with diverse targets involved in the pathogenesis underlying AD

Loss of mitochondrial Ndufs4 in striatal medium spiny neurons mediates progressive motor impairment in a mouse model of leigh syndrome

Altres ajuts: Juan del la Cierva (IJCI-2015-24576)Inability of mitochondria to generate energy leads to severe and often fatal myoencephalopathies. Among these, Leigh syndrome (LS) is one of the most common childhood mitochondrial diseases; it is characterized by hypotonia, failure to thrive, respiratory insufficiency and progressive mental and motor dysfunction, leading to early death. Basal ganglia nuclei, including the striatum, are affected in LS patients. However, neither the identity of the affected cell types in the striatum nor their contribution to the disease has been established. Here, we used a mouse model of LS lacking Ndufs4, a mitochondrial complex I subunit, to confirm that loss of complex I, but not complex II, alters respiration in the striatum. To assess the role of striatal dysfunction in the pathology, we selectively inactivated Ndufs4 in the striatal medium spiny neurons (MSNs), which account for over 95% of striatal neurons. Our results show that lack of Ndufs4 in MSNs causes a non-fatal progressive motor impairment without affecting the cognitive function of mice. Furthermore, no inflammatory responses or neuronal loss were observed up to 6 months of age. Hence, complex I deficiency in MSNs contributes to the motor deficits observed in LS, but not to the neural degeneration, suggesting that other neuronal populations drive the plethora of clinical signs in LS

Multi-Target Directed Donepezil-Like Ligands for Alzheimer's Disease

HIGHLIGHTS Alzheimer's disease (AD), the most common form of adult onset dementia, is an age-related neurodegenerative disorder characterized by progressive memory loss, decline in language skills, and other cognitive impairments. Although its etiology is not completely known, several factors including deficits of acetylcholine, β-amyloid deposits, τ-protein phosphorylation, oxidative stress, and neuroinflammation are considered to play significant roles in the pathophysiology of this disease. For a long time, AD patients have been treated with acetylcholinesterase inhibitors such as donepezil (Aricept®) but with limited therapeutic success. This might be due to the complex multifactorial nature of AD, a fact that has prompted the design of new Multi-Target-Directed Ligands (MTDL) based on the "one molecule, multiple targets" paradigm. Thus, in this context, different series of novel multifunctional molecules with antioxidant, anti-amyloid, anti-inflammatory, and metal-chelating properties able to interact with multiple enzymes of therapeutic interest in AD pathology including acetylcholinesterase, butyrylcholinesterase, and monoamine oxidases A and B have been designed and assessed biologically. This review describes the multiple targets, the design rationale and an in-house MTDL library, bearing the N -benzylpiperidine motif present in donepezil, linked to different heterocyclic ring systems (indole, pyridine, or 8-hydroxyquinoline) with special emphasis on compound ASS234, an N -propargylindole derivative. The description of the in vitro biological properties of the compounds and discussion of the corresponding structure-activity-relationships allows us to highlight new issues for the identification of more efficient MTDL for use in AD therapy