Uso del 3-(2-isotiocianatoetil)-5-metoxi-1H-indol para el tratamiento de enfermedades neurodegenerativas

La presente invención se refiere al uso del 3-(2- isotiocianatoetil)-5-metoxi-1H-indol o de una composición que comprende el 3-(2-isotiocianatoetil)- 5-metoxi-1H-indol para el tratamiento de enfermedades que cursan con declive de la capacidad cognitiva o motoras secundarias a degeneración neuronal. La presente invención también se refiere al uso del 3-(2-isotiocianatoetil)-5- metoxi-1H-indol para el tratamiento de otras enfermedades neurodegenerativas que cursan con elevados niveles de estrés oxidativo y/o neuroinflamación y/o autoinmunes. Preferiblemente, la presente invención se refiere al uso del 3-(2- isotiocianatoetil)-5-metoxi-1H-indol para el tratamiento de la enfermedad de Alzheimer, la enfermedad de Parkinson, la enfermedad de Huntington, esclerosis múltiple, ictus cerebral o esclerosis lateral amiotrófica.Peer reviewedFundación para la Investigación Biomédica del Hospital Universitario de La Princesa, Universidad Autónoma de Madrid, Consejo Superior de Investigaciones Científicas, DNS Neuroscience S.A.B1 Patente sin examen previ

The melatonin- N,N-dibenzyl(N-methyl)amine hybrid ITH91/IQM157 affords neuroprotection in an in vitro Alzheimer's model via hemo-oxygenase-1 induction

We have investigated the protective effects of ITH91/IQM157, a hybrid of melatonin and N,N-dibenzyl(N-methyl)amine, in an in vitro model of Alzheimer's disease (AD)-like pathology that combines amyloid beta (Aβ) and tau hyperphosphorylation induced by okadaic acid (OA), in the human neuroblastoma cell line SH-SY5Y. Combination of subtoxic concentrations of Aβ and OA caused a significant toxicity of 40% cell death, which mainly was apoptotic; this effect was accompanied by retraction of the cells' prolongations and accumulation of thioflavin-S stained protein aggregates. In this toxicity model, ITH91/IQM157 (1-1000 nM) reduced cell death measured as MTT reduction; at 100 nM, it prevented apoptosis, retraction of prolongations, and Aβ aggregates. The protective actions of ITH91/IQM157 were blocked by mecamylamine, luzindol, chelerythrine, PD98059, LY294002, and SnPP. We show that the combination of melatonin with a fragment endowed with AChE inhibition in a unique chemical structure, ITH91/IQM157, can reduce neuronal cell death induced by Aβ and OA by a signaling pathway that implicates both nicotinic and melatonin receptors, PKC, Akt, ERK1/2, and induction of hemoxygenase-1.Peer Reviewe

Agmatine, by improving neuroplasticity markers and inducing Nrf2, prevents corticosterone-induced depressive-like behavior in mice

Agmatine, an endogenous neuromodulator, is a potential candidate to constitute an adjuvant/monotherapy for the management of depression. A recent study by our group demonstrated that agmatine induces Nrf2 and protects against corticosterone effects in a hippocampal neuronal cell line. The present study is an extension of this previous study by assessing the antidepressant-like effect of agmatine in an animal model of depression induced by corticosterone in mice. Swiss mice were treated simultaneously with agmatine or imipramine at a dose of 0.1 mg/kg/day (p.o.) and corticosterone for 21 days and the daily administrations of experimental drugs were given immediately prior to corticosterone (20 mg/kg/day, p.o.) administrations. Wild-type C57BL/6 mice (Nrf2 (+/+)) and Nrf2 KO (Nrf2 (−/−)) were treated during 21 days with agmatine (0.1 mg/kg/day, p.o.) or vehicle. Twenty-four hours after the last treatments, the behavioral tests and biochemical assays were performed. Agmatine treatment for 21 days was able to abolish the corticosterone-induced depressive-like behavior and the alterations in the immunocontent of mature BDNF and synaptotagmin I, and in the serotonin and glutamate levels. Agmatine also abolished the corticosterone-induced changes in the morphology of astrocytes and microglia in CA1 region of hippocampus. In addition, agmatine treatment in control mice increased noradrenaline, serotonin, and dopamine levels, CREB phosphorylation, mature BDNF and synaptotagmin I immunocontents, and reduced pro-BDNF immunocontent in the hippocampus. Agmatine’s ability to produce an antidepressant-like effect was abolished in Nrf2 (−/−) mice. The present results reinforce the participation of Nrf2 in the antidepressant-like effect produced by agmatine and expand literature data concerning its mechanisms of action.This work was supported by CNPq, CAPES/PDSE, CAPES/PROCAD, Núcleo de Excelência em Neurociências Aplicadas de Santa Catarina (NENASC) Project/ PRONEX Program CNPq/FAPESC (Brazil) to ALSR. The 8th Convocatoria de proyectos de Cooperación Interuniversitaria UAM-Santander con America Latina, Spanish Ministry of Economy and Competence Ref. SAF2012-32223 to MGL and ALSR. This work was also supported by FIS CA12/00122 to ARN and FPU12/02220 to AW.Peer Reviewe

Melatonin-sulforaphane hybrid ITH12674 attenuates glial response in vivo by blocking LPS binding to MD2 and receptor oligomerization

Neuroinflammation is increasingly associated to the onset and progression of neurodegenerative diseases. Furthermore, several lines of evidence have demonstrated the capacity of aberrant protein aggregates to activate the immune response, accelerating the advance of the disease. Compound ITH12674 is a melatonin-sulforaphane hybrid designed to exert a dual drug-prodrug mechanism of action that combines potent NRF2 induction and free radical scavenger activity. ITH12674 also showed neuroprotective properties in oxidative stress related models, that were dependant on its NRF2 inducing properties. Given the high impact of neuroinflammation in the pathogenesis of neurodegeneration, we foresaw to study the anti-inflammatory properties of ITH12674. ITH12674 reduced inflammatory markers in glial cell cultures and hippocampal tissue after LPS administration. The anti-inflammatory effect was related to inhibition of TLR4 receptors due to a direct interaction with the TLR4/MD2 complex at the hydrophobic cavity of MD2. ITH12674 is endowed with anti-inflammatory properties, that are complementary to the NRF2 inducing activity and neuroprotective properties. Thus, ITH12674 could be of potential interest for the treatment of diseases with chronic neuroinflammation.This work was supported by grants from IS Carlos III co-financed by the European Regional Development’s funds (FEDER), Programa Miguel Servet II (CP16/00014) and research project (grant PI17/01700) to RL; Fundación La Caixa, CaixaImpulse program (grant CI17-00048) to RL; Comunidad Autónoma de Madrid (grant B2017/BMD-3827) and Spanish Ministry of Science, Innovation and Education (grant RTI2018-095793-B-I00) to MGL. P.M., P.D. and C.F.-M thanks MECD for FPU fellowships (13/0373716/03977 and 15/03269 respectively). I.B thanks MECD for Juan de la Cierva fellowship (FJCI-2016/28282). We would also like to thank “Fundación Teófilo Hernando” for its continued support.Peer reviewe

Therapeutic agents for the treatment of neurodegenerative diseases: Nrf2 as central player

Resumen del póster presentado al 3rd Symposium on Biomedical Research: "Advances and Perspectives in Neuroscience", celebrado en la Universidad Autónoma de Madrid el 22 de abril de 2016.The continuous aging of population has led to an ever-increasing number of people afflicted by neurodegenerative diseases (NDDs). NDDs share many pathophysiological pathways, such as mitochondrial dysfunction, aberrant protein aggregation, oxidative stress and neuroinflammation. Although the multifactorial nature of NDDs hinders the development of efficient treatments, there is a great interest in the development of compounds directed to reduce oxidative stress and neuroinflammation as these alterations play a crucial role in the evolution of those pathologies. The Nrf2 pathway plays a central role in the defense against both, oxidative stress and neuroinflammation. Besides, this pathway is deregulated in NDDs. Herein, we propose the design, synthesis, and pharmacological evaluation of new compounds, hybrids of melatonin (potent antioxidant) and cinnamic acid (moderate Nrf2 inducer), as potential treatments for NDDs. The pharmacological screening of newly developed compounds showed the compounds are Nrf2 inducers, potent scavengers and anti-inflammatory agents. Besides, these compounds have demonstrated an interesting neuroprotective profile against different in vitro models, and a broad security profile. Furthermore, compound 11 has demonstrated a potent neurogenic effect via melatonin receptors and Nrf2 induction. Altogether, these results indicate these compounds should be further evaluated in in vivo models of NDDs.Peer reviewe

Deficiency in the transcription factor NRF2 worsens inflammatory parameters in a mouse model with combined tauopathy and amyloidopathy

Chronic neuroinflammation is a hallmark of the onset and progression of brain proteinopathies such as Alzheimer disease (AD) and it is suspected to participate in the neurodegenerative process. Transcription factor NRF2, a master regulator of redox homeostasis, controls acute inflammation but its relevance in low-grade chronic inflammation of AD is inconclusive due to lack of good mouse models. We have addressed this question in a transgenic mouse that combines amyloidopathy and tauopathy with either wild type (AT-NRF2-WT) or NRF2-deficiency (AT-NRF2-KO). AT-NRF2-WT mice died prematurely, at around 14 months of age, due to motor deficits and a terminal spinal deformity but AT-NRF2-KO mice died roughly 2 months earlier. NRF2-deficiency correlated with exacerbated astrogliosis and microgliosis, as determined by an increase in GFAP, IBA1 and CD11b levels. The immunomodulatory molecule dimethyl fumarate (DMF), a drug already used for the treatment of multiple sclerosis whose main target is accepted to be NRF2, was tested in this preclinical model. Daily oral gavage of DMF during six weeks reduced glial and inflammatory markers and improved cognition and motor complications in the AT-NRF2-WT mice compared with the vehicle-treated animals. This study demonstrates the relevance of the inflammatory response in experimental AD, tightly regulated by NRF2 activity, and provides a new strategy to fight AD.This work was funded by SAF2016-76520-R of the Spanish Ministry of Economy and Competitiveness, a Pathfinder grant of the Centres of Excellence in Neurodegeneration (COEN) of the EU-Joint Program for Neurodegenerative diseases, and European Regional Development Fund, Competitiveness Operational Program 2014–2020, through the grant P_37_732/2016 REDBRAIN and a collaborative CIBERNED project PI2017/04-3. M.P. is recipient of a FPU fellowship of Autonomous University of Madrid.Peer reviewe

Deficiency in the transcription factor NRF2 worsens inflammatory parameters in a mouse model with combined tauopathy and amyloidopathy

Resumen del trabajo presentado al 5th Symposium on Biomedical Research: "Advances and Perspectives In Pharmacology, Drug Toxicity and Pharmacogenetics", celebrado en Madrid del 15 al 16 de marzo de 2018.Chronic neuroinflammation is a hallmark of the onset and progression of brain proteinopathies such as Alzheimer disease (AD) and it is suspected to participate in the neurodegenerative process. Transcription factor NRF2, a master regulator of redox homeostasis, controls acute inflammation but its relevance in low-grade chronic inflammation of AD is inconclusive due to lack of good mouse models. We have addressed this question in a transgenic mouse that combines amyloidopathy and tauopathy with either wild type (AT-NRF2-WT) or NRF2-deficiency (AT-NRF2-KO). AT-NRF2-WT mice died prematurely, at around 14 months of age, due to motor deficits and a terminal spinal deformity but AT-NRF2-KO mice died roughly 2 months earlier. NRF2-deficiency correlated with exacerbated astrogliosis and microgliosis, as determined by an increase in GFAP, IBA1 and CD11b levels. The immunomodulatory molecule dimethyl fumarate (DMF), a drug already used for the treatment of multiple sclerosis whose main target is accepted to be NRF2, was tested in this preclinical model. Daily oral gavage of DMF during six weeks reduced glial and inflammatory markers and improved cognition and motor complications in the AT-NRF2-WT mice compared with the vehicle-treated animals. This study demonstrates the relevance of the inflammatory response in experimental AD, tightly regulated by NRF2 activity, and provides a new strategy to fight AD.Peer Reviewe

Multitarget compounds for the treatment of neurodegenerative diseases: Nrf2-EpRE pathway as key target

Resumen del trabajo presentado al 1st PhD Research Symposium in Health Sciences and Biomedicine, celebrado en la Universidad Autónoma de Madrid el 18 de mayo de 2018.IS Carlos III (Contrato Miguel Servet II a RL, proyecto CPII16/00014 y proyecto PI14/00372 a RL),). Ministerio de Educación, Cultura y Deporte (MECD) FPU13/03737 a P.M, FPU16/03977 a P.D.Peer reviewe

Discovery of the first dual GSK3β inhibitor/Nrf2 inducer. A new multitarget therapeutic strategy for Alzheimer's disease

The formation of neurofibrillary tangles (NFTs), oxidative stress and neuroinflammation have emerged as key targets for the treatment of Alzheimer's disease (AD), the most prevalent neurodegenerative disorder. These pathological hallmarks are closely related to the over-activity of the enzyme GSK3β and the downregulation of the defense pathway Nrf2-EpRE observed in AD patients. Herein, we report the synthesis and pharmacological evaluation of a new family of multitarget 2,4-dihydropyrano[2,3-c]pyrazoles as dual GSK3β inhibitors and Nrf2 inducers. These compounds are able to inhibit GSK3β and induce the Nrf2 phase II antioxidant and anti-inflammatory pathway at micromolar concentrations, showing interesting structure-activity relationships. The association of both activities has resulted in a remarkable anti-inflammatory ability with an interesting neuroprotective profile on in vitro models of neuronal death induced by oxidative stress and energy depletion and AD. Furthermore, none of the compounds exhibited in vitro neurotoxicity or hepatotoxicity and hence they had improved safety profiles compared to the known electrophilic Nrf2 inducers. In conclusion, the combination of both activities in this family of multitarget compounds confers them a notable interest for the development of lead compounds for the treatment of AD.R.L. thanks Instituto de Salud Carlos III co-financed by the European Regional Development's funds (FEDER) for a research contract under Miguel Servet Program (CP11/00165) and financial support from the European Commission-REA, People (Marie Curie Actions) FP7 under REA grant agreement n° PCIG11-GA-2012-322156; Spanish Ministry of Health (Instituto de Salud Carlos III) (grant PI14/00372); Bayer AG, From Targets to Drugs (grant 2015-03-1282) and Fundación FIPSE (grant 12-00001344-15). Ministerio de Economía y Competitividad, MINECO (grant CTQ-2015-68380-R) to J.C.M and grant SAF2015-63935R to M.G.L.; GW thanks Cancer Research UK (grant C9344/A10268), the BBSRC (grant BB/L01923X/1), the Bloomsbury Consortium and UCL School of Pharmacy for financial support. G.T. thanks Universidad Complutense for a predoctoral fellowship. P.M. thanks MECD for FPU fellowship (FPU13/03737). I. G. thanks UAM for a FPI fellowship. We also gratefully acknowledge the continued support of Instituto-Fundación Teófilo Hernando, Madrid, Spain.Peer Reviewe

Heme-oxygenase I and PCG-1α regulate mitochondrial biogenesis via microglial activation of alpha7 nicotinic acetylcholine receptors using PNU282987

[Aims]: A loss in brain acetylcholine and cholinergic markers, subchronic inflammation, and impaired mitochondrial function, which lead to low-energy production and high oxidative stress, are common pathological factors in several neurodegenerative diseases (NDDs). Glial cells are important for brain homeostasis, and microglia controls the central immune response, where α7 acetylcholine nicotinic receptors (nAChR) seem to play a pivotal role; however, little is known about the effects of this receptor in metabolism. Therefore, the aim of this study was to evaluate if glial mitochondrial energetics could be regulated through α7 nAChR. [Results]: Primary glial cultures treated with the α7 nicotinic agonist PNU282987 increased their mitochondrial mass and their mitochondrial oxygen consumption without increasing oxidative stress; these changes were abolished when nuclear erythroid 2-related factor 2 (Nrf2) was absent, heme oxygenase-1 (HO-1) was inhibited, or peroxisome proliferator-activated receptor gamma coactivator 1α (PGC-1α) was silenced. More specifically, microglia of animals treated intraperitoneally with the α7 nAChR agonist PNU282987 (10 mg/kg) showed a significant increase in mitochondrial mass. Interestingly, LysMcre-Hmox1Δ/Δ and PGC-1α−/− animals showed lower microglial mitochondrial levels and treatment with PNU282987 did not produce effects on mitochondrial levels.[Innovation]: Increases in microglial mitochondrial mass and metabolism can be achieved via α7 nAChR by a mechanism that implicates Nrf2, HO-1, and PGC-1α. This signaling pathway could open a new strategy for the treatment of NDDs, such as Alzheimer's, characterized by a reduction of cholinergic markers. [Conclusion]: α7 nAChR signaling increases glial mitochondrial mass, both in vitro and in vivo, via HO-1 and PCG-1α. These effects could be of potential benefit in the context of NDDs.This work was supported by the Spanish Ministry of Economy and Competence Ref. SAF2012-23332 and SAF2015-63935-R to M.G.L.; Ref. SAF2012-37693 to M.M. Comunidad Autónoma de Madrid grant number S2010/BMD-2361 to M.M. E.N. a predoctoral fellowship from Universidad Autónoma de Madrid. The work in SC laboratory is supported by grants from the Instituto de Salud Carlos III FIS (PI12/00933 and PI15/00448) and by Comunidad de Madrid (S2011/BMD-2402). We also thank the Fundación Teófilo Hernando, the Network of Excellence Nrf2-net of MINECO, and the European Cooperation and Technology (COST Action BM1203/EU-ROS).Peer reviewe