Subscriber access provided by UNIV OF DURHAM
Letter
Neuroinflammation signaling modulated by ASS234, a multitarget small molecule for Alzheimer disease therapy Javier Del Pino, José Marco-Contelles, Francisco Lopez-Muñoz, Alejandro Romero, and Eva Ramos ACS Chem. Neurosci., Just Accepted Manuscript • DOI: 10.1021/acschemneuro.8b00203 • Publication Date (Web): 26 Jul 2018 Downloaded from http://pubs.acs.org on July 27, 2018
Just Accepted “Just Accepted” manuscripts have been peer-reviewed and accepted for publication. They are posted online prior to technical editing, formatting for publication and author proofing. The American Chemical Society provides “Just Accepted” as a service to the research community to expedite the dissemination of scientific material as soon as possible after acceptance. “Just Accepted” manuscripts appear in full in PDF format accompanied by an HTML abstract. “Just Accepted” manuscripts have been fully peer reviewed, but should not be considered the official version of record. They are citable by the Digital Object Identifier (DOI®). “Just Accepted” is an optional service offered to authors. Therefore, the “Just Accepted” Web site may not include all articles that will be published in the journal. After a manuscript is technically edited and formatted, it will be removed from the “Just Accepted” Web site and published as an ASAP article. Note that technical editing may introduce minor changes to the manuscript text and/or graphics which could affect content, and all legal disclaimers and ethical guidelines that apply to the journal pertain. ACS cannot be held responsible for errors or consequences arising from the use of information contained in these “Just Accepted” manuscripts.
is published by the American Chemical Society. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society. Copyright © American Chemical Society. However, no copyright claim is made to original U.S. Government works, or works produced by employees of any Commonwealth realm Crown government in the course of their duties.
Page 1 of 16 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
ACS Chemical Neuroscience
Neuroinflammation signaling modulated by ASS234, a multitarget small molecule for Alzheimer disease therapy †
‡
Javier del Pino, José Marco-Contelles, Francisco López-Muñoz, Ramos †
#,&
Alejandro Romero,
†,*
and Eva
†,*
Department of Pharmacology and Toxicology, Faculty of Veterinary Medicine, Complutense University of Madrid, Madrid, Spain. ‡
Laboratory of Medicinal Chemistry, Institute of General Organic Chemistry (CSIC) Madrid, Spain.
#
Faculty of Health, Camilo José Cela University, Villanueva de la Cañada, Madrid, Spain.
&
Neuropsychopharmacology Unit, “Hospital 12 de Octubre” Research Institute, Madrid, Spain.
ABSTRACT: There are clear evidences that neuroinflammation plays a crucial role in the pathogenesis of Alzheimer's disease. Consequently, modulating the inflammatory environment in brain has become a powerful and attractive strategy to deal with Alzheimer's disease physiopathology. In spite of the neuroprotective capacity shown by ASS234, a multitarget propargylamine targeted for Alzheimer's disease, its regulation of inflammation in the brain still remains unexplored. Therefore, we aimed to characterize ASS234 possible anti-inflammatory effects, counteracting induced inflammatory effects in RAW 264.7 cells and evaluating seven neuroinflammation related genes expression profiling (IL-6, IL-10, IL1ß, NF-κB, TNF-α, TNFR1, and TGF-ß), after ASS234 (5µM) treatment in SH-SY5Y cells. The analysis of the obtained fold changes lead us to conclude that ASS234 may play an important role facing the neuroinflammatory environment in Alzheimer's disease pathology.
KEYWORDS: Alzheimer disease, ASS234, neuroinflammation, multitarget small compounds, gene expression, signaling pathways
ACS Paragon Plus Environment
ACS Chemical Neuroscience 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
Alzheimer's disease (AD) is a multifactorial neurodegenerative disorder, characterized by strong cognitive impairment that finally leads to death. Extensive neuroinflammation in AD patients may exacerbate its pathogenesis. Therefore, therapies directed to reduce the inflammation process in brain may lessen AD progress and development.1-3 However, not many therapeutic tools against this disease are available. For this reason, our group has demonstrated a strong commitment in the search of new molecules against neurodegeneration. In this context N-((5-(3-(1-benzylpiperidin-4-yl) propoxy) -1-methyl-1H-indol-2-yl) methyl) -N-methylprop-2-yn-1-amine (ASS234, Figure 1), has been synthesized as a new multitarget direct small compound potentially effective for AD. ASS234 is able to cross blood brain barrier.4 Several therapeutic activities and its action mechanisms had been previously described.5-11 Regardless, its activity and safety profile should be more deeply investigated, specifically in what concerns to its anti-neuroinflammation capacity. FIGURE 1 Figure 1. Chemical structures of ASS234 and ladostigil, highlighting common functional motifs as the possible responsible feature of their anti-inflammatory effects. Recent reports, had shown that the anti-inflammatory effects displayed by a number of Multi Target Directed Ligands (MTDLs) correlate very well with their capacity to inhibit GSK-3β enzymes.12-14 In addition, its association with the induction of the Nrf-2,12 or β-secretase inhibition, is a promising new therapeutic approach for AD therapy.13-14 We can hypothesize ASS234 anti-inflammatory effects from a structural point of view, by a simple comparison with ladostigil15 (Figure 1). Antiinflammatory activity can be attributed to the presence of a common N-propargylamine moiety in both compounds, responsible inter alia of their remarkable neuroprotective properties.4, 16 Therefore, we herein performed a gene analysis study to investigate its possible anti-inflammatory action mechanism.
Besides the classical in AD hallmarks, extracellular amyloid plaques (Aß) and intracellular deposits of neurofibrillary tangles (NFTs), microglia plays a significant role in the neurodegenerative process. Microglia has been described as the immune surveillance of the central nervous system (CNS) to maintain its homeostasis. Nevertheless, its over-activation leads to the release of pro-inflammatory factors, such as, IL-6, IL-1ß, TNF-α, contributing to AD progress.17-18 Moreover, Aß and NFTs are exacerbated in the presence of pro-inflammatory cytokines.3, 17 Chronic neuroinflammation, indeed, has been widely described as a deterioration co-factor in AD, and chronic overexpression of proinflammatory cytokines worsens AD progression.1-3, 19 Taking into account the complexity of the AD
ACS Paragon Plus Environment
Page 2 of 16
Page 3 of 16 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
ACS Chemical Neuroscience
brain environment, targeting pro-inflammatory factors and AD engaged signaling pathways are promising therapeutic approaches.
In previous studies, we have evaluated Wnt and Nrf-2 signaling pathways as possible neuroprotection mechanisms, showing that ASS234 upregulated them eliciting a neuroprotective profile.8-9 These pathways have been related to CNS inflammation, thus in order to deepen the neuroprotective profile of ASS234, we evaluated its possible effects related to the inflammatory response in SH-SY5Y cells. As a first step, ASS234 was tested in macrophage RAW 264.7 cell line, stimulated with Lipopolysaccharide (LPS) to simulate an inflammatory environment. ASS234 was able to inhibit in a statistically significant manner the inflammatory response in LPS-stimulated RAW 264.7 macrophages after 24 h co-treatment (Figure 2). While it is true that MTT assay does not completely justify the anti-inflammatory effect of ASS234, it is an additional test that supports the initial hypothesis, since this inflammation induced environment was partially counteracted
FIGURE 2 Figure 2. Anti-inflammatory effect of ASS234 (5 µM) in LPS-stimulated RAW 264.7 cells measured through the MTT assay. Data correspond to the mean ± S.E.M. of four independent experiments in triplicate. ***p< 0.001 respect to basal, &&p
