Garrido J

References (11)

Title : Fine-Tuning the Biological Profile of Multitarget Mitochondriotropic Antioxidants for Neurodegenerative Diseases - Chavarria_2021_Antioxidants.(Basel)_10_329
Author(s) : Chavarria D , Da Silva O , Benfeito S , Barreiro S , Garrido J , Cagide F , Soares P , Remiao F , Brazzolotto X , Nachon F , Oliveira PJ , Dias J , Borges F
Ref : Antioxidants (Basel) , 10 :329 , 2021
Abstract : Neurotransmitter depletion and mitochondrial dysfunction are among the multiple pathological events that lead to neurodegeneration. Following our previous studies related with the development of multitarget mitochondriotropic antioxidants, this study aims to evaluate whether the Pi-system extension on the chemical scaffolds of AntiOXCIN2 and AntiOXCIN3 affects their bioactivity and safety profiles. After the synthesis of four triphenylphosphonium (TPP(+)) conjugates (compounds 2-5), we evaluated their antioxidant properties and their effect on neurotransmitter-metabolizing enzymes. All compounds were potent equine butyrylcholinesterase (eqBChE) and moderate electric eel acetylcholinesterase (eeAChE) inhibitors, with catechols 4 and 5 presenting lower IC(50) values than AntiOXCIN2 and AntiOXCIN3, respectively. However, differences in the inhibition potency and selectivity of compounds 2-5 towards non-human and human cholinesterases (ChEs) were observed. Co-crystallization studies with compounds 2-5 in complex with human ChEs (hChEs) showed that these compounds exhibit different binging modes to hAChE and hBChE. Unlike AntiOXCINs, compounds 2-5 displayed moderate human monoamine oxidase (hMAO) inhibitory activity. Moreover, compounds 4 and 5 presented higher ORAC-FL indexes and lower oxidation potential values than the corresponding AntiOXCINs. Catechols 4 and 5 exhibited broader safety windows in differentiated neuroblastoma cells than benzodioxole derivatives 2 and 3. Compound 4 is highlighted as a safe mitochondria-targeted antioxidant with dual ChE/MAO inhibitory activity. Overall, this work is a contribution for the development of dual therapeutic agents addressing both mitochondrial oxidative stress and neurotransmitter depletion.
ESTHER : Chavarria_2021_Antioxidants.(Basel)_10_329
PubMedSearch : Chavarria_2021_Antioxidants.(Basel)_10_329
PubMedID: 33672269
Gene_locus related to this paper: human-ACHE , human-BCHE

Title : Benzoic acid-derived nitrones: A new class of potential acetylcholinesterase inhibitors and neuroprotective agents - Oliveira_2019_Eur.J.Med.Chem_174_116
Author(s) : Oliveira C , Bagetta D , Cagide F , Teixeira J , Amorim R , Silva T , Garrido J , Remiao F , Uriarte E , Oliveira PJ , Alcaro S , Ortuso F , Borges F
Ref : Eur Journal of Medicinal Chemistry , 174 :116 , 2019
Abstract : The discovery of new chemical entities endowed with potent and selective acetylcholinesterase (AChE) and/or butyrylcholinesterase (BChE) inhibitory activity is still a relevant subject for Alzheimer's disease therapy. Therefore, a small library of benzoic based amide nitrones (compounds 24 to 42) was synthesized and screened toward cholinesterase enzymes. SAR studies showed that the tert-butyl moiety is the most favourable nitrone pattern. In general, tert-butyl derivatives effectively inhibited AChE, being compound 33 the most potent (IC50=8.3+/-0.3muM; Ki 5.2muM). The data pointed to a non-competitive inhibition mechanism of action, which was also observed for the standard donepezil. None of compounds showed BChE inhibitory activity. Molecular modelling studies provided insights into the enzyme-inhibitor interactions and rationalised the experimental data, confirming that the binding mode of nitrones 33 and 38 towards AChE has the most favourable binding free energy. The tert-butylnitrones 33 and 38 were not cytotoxic on different cell lines (SH-SY5Y and HepG2). Moreover, compound 33 was able to prevent t-BHP-induced oxidative stress in SH-SY5Y differentiated cells. Due to its AChE selectivity and promising cytoprotective properties, as well as its appropriate drug-like profile pointing toward blood-brain barrier permeability, compound 33 is proposed as a valid lead for a further optimization step.
ESTHER : Oliveira_2019_Eur.J.Med.Chem_174_116
PubMedSearch : Oliveira_2019_Eur.J.Med.Chem_174_116
PubMedID: 31029943

Title : Hydroxybenzoic Acid Derivatives as Dual-Target Ligands: Mitochondriotropic Antioxidants and Cholinesterase Inhibitors - Oliveira_2018_Front.Chem_6_126
Author(s) : Oliveira C , Cagide F , Teixeira J , Amorim R , Sequeira L , Mesiti F , Silva T , Garrido J , Remiao F , Vilar S , Uriarte E , Oliveira PJ , Borges F
Ref : Front Chem , 6 :126 , 2018
Abstract : Alzheimer's disease (AD) is a multifactorial age-related disease associated with oxidative stress (OS) and impaired cholinergic transmission. Accordingly, targeting mitochondrial OS and restoring cholinergic transmission can be an effective therapeutic strategy toward AD. Herein, we report for the first time dual-target hydroxybenzoic acid (HBAc) derivatives acting as mitochondriotropic antioxidants and cholinesterase (ChE) inhibitors. The studies were performed with two mitochondriotropic antioxidants AntiOxBEN1 (catechol derivative), and AntiOxBEN2 (pyrogallol derivative) and compounds 15-18, which have longer spacers. Compounds AntiOxBEN1 and 15, with a shorter carbon chain spacer (six- and eight-carbon) were shown to be potent antioxidants and BChE inhibitors (IC50 = 85 +/- 5 and 106 +/- 5 nM, respectively), while compounds 17 and 18 with a 10-carbon chain were more effective AChE inhibitors (IC50 = 7.7 +/- 0.4 and 7.2 +/- 0.5 muM, respectively). Interestingly, molecular modeling data pointed toward bifunctional ChEs inhibitors. The most promising ChE inhibitors acted by a non-competitive mechanism. In general, with exception of compounds 15 and 17, no cytotoxic effects were observed in differentiated human neuroblastoma (SH-SY5Y) and human hepatocarcinoma (HepG2) cells, while Abeta-induced cytotoxicity was significantly prevented by the new dual-target HBAc derivatives. Overall, due to its BChE selectivity, favorable toxicological profile, neuroprotective activity and drug-like properties, which suggested blood-brain barrier (BBB) permeability, the mitochondriotropic antioxidant AntiOxBEN1 is considered a valid lead candidate for the development of dual acting drugs for AD and other mitochondrial OS-related diseases.
ESTHER : Oliveira_2018_Front.Chem_6_126
PubMedSearch : Oliveira_2018_Front.Chem_6_126
PubMedID: 29740575

Title : Effects of chlorophenoxy herbicides and their main transformation products on DNA damage and acetylcholinesterase activity - Benfeito_2014_Biomed.Res.Int_2014_709036
Author(s) : Benfeito S , Silva T , Garrido J , Andrade PB , Sottomayor MJ , Borges F , Garrido EM
Ref : Biomed Res Int , 2014 :709036 , 2014
Abstract : Persistent pesticide transformation products (TPs) are increasingly being detected among different environmental compartments, including groundwater and surface water. However, there is no sufficient experimental data on their toxicological potential to assess the risk associated with TPs, even if their occurrence is known. In this study, the interaction of chlorophenoxy herbicides (MCPA, mecoprop, 2,4-D and dichlorprop) and their main transformation products with calf thymus DNA by UV-visible absorption spectroscopy has been assessed. Additionally, the toxicity of the chlorophenoxy herbicides and TPs was also assessed evaluating the inhibition of acetylcholinesterase activity. On the basis of the results found, it seems that AChE is not the main target of chlorophenoxy herbicides and their TPs. However, the results found showed that the transformation products displayed a higher inhibitory activity when compared with the parent herbicides. The results obtained in the DNA interaction studies showed, in general, a slight effect on the stability of the double helix. However, the data found for 4-chloro-2-methyl-6-nitrophenol suggest that this transformation product can interact with DNA through a noncovalent mode.
ESTHER : Benfeito_2014_Biomed.Res.Int_2014_709036
PubMedSearch : Benfeito_2014_Biomed.Res.Int_2014_709036
PubMedID: 24795892

Title : A major portion of synaptic basal lamina acetylcholinesterase is detached by high salt- and heparin-containing buffers from rat diaphragm muscle and torpedo electric organ - Casanueva_1998_J.Biol.Chem_273_4258
Author(s) : Casanueva OI , Garcia-Huidobro T , Campos EO , Aldunate R , Garrido J , Inestrosa NC
Ref : Journal of Biological Chemistry , 273 :4258 , 1998
Abstract : Collagen-tailed asymmetric acetylcholinesterase (AChE) forms are believed to be anchored to the synaptic basal lamina via electrostatic interactions involving proteoglycans. However, it was recently found that in avian and rat muscles, high ionic strength or polyanionic buffers could not detach AChE from cell-surface clusters and that these buffers solubilized intracellular non-junctional asymmetric AChE rather than synaptic forms of the enzyme. In the present study, asymmetric AChE forms were specifically solubilized by ionic buffers from synaptic basal lamina-enriched fractions, largely devoid of intracellular material, obtained from the electric organ of Torpedo californica and the end plate regions of rat diaphragm muscle. Furthermore, foci of AChE activity were seen to diminish in size, number, and staining intensity when the rat synaptic basal lamina-enriched preparations were treated with the extraction buffers. In the case of Torpedo, almost all the AChE activity was removed from the pure basal lamina sheets. We therefore conclude that a major portion of extracellular collagen-tailed AChE is extractable from rat and Torpedo synaptic basal lamina by high ionic strength and heparin buffers, although some non-extractable AChE activity remains associated with the junctional regions.
ESTHER : Casanueva_1998_J.Biol.Chem_273_4258
PubMedSearch : Casanueva_1998_J.Biol.Chem_273_4258
PubMedID: 9461624

Title : A monoclonal antibody against acetylcholinesterase inhibits the formation of amyloid fibrils induced by the enzyme - Reyes_1997_Biochem.Biophys.Res.Commun_232_652
Author(s) : Reyes AE , Perez DR , Alvarez A , Garrido J , Gentry MK , Doctor BP , Inestrosa NC
Ref : Biochemical & Biophysical Research Communications , 232 :652 , 1997
Abstract : A monoclonal antibody (mAb) 25B1 directed against fetal bovine-serum acetylcholinesterase (FBS AChE) was used to examine the ability of the cholinergic enzyme to promote the assembly of amyloid-beta peptides (A beta) into Alzheimer fibrils. This mAb binds to the peripheral anionic site of the enzyme and allosterically inhibits catalytic activity of FBS AChE. Several techniques, including thioflavine-T fluorescence, turbidity, and negative-staining at the electron microscopy level, were used to assess amyloid formation. Inhibition of amyloid formation was dependent on the molar ratio AChE:mAb 25B1, and at least 50% of the inhibition of the AChE promoting effect occurs at a molar ratio similar to that required for inhibition of the esterase activity. Our results suggest that mAb 25B1 inhibits the promotion of the amyloid fibril formation triggered by AChE by affecting the lag period of the A beta aggregation process.
ESTHER : Reyes_1997_Biochem.Biophys.Res.Commun_232_652
PubMedSearch : Reyes_1997_Biochem.Biophys.Res.Commun_232_652
PubMedID: 9126330

Title : Acetylcholinesterase promotes the aggregation of amyloid-beta-peptide fragments by forming a complex with the growing fibrils - Alvarez_1997_J.Mol.Biol_272_348
Author(s) : Alvarez A , Opazo C , Alarcon R , Garrido J , Inestrosa NC
Ref : Journal of Molecular Biology , 272 :348 , 1997
Abstract : Acetylcholinesterase (AChE), an enzyme involved in the hydrolysis of the neurotransmitter acetylcholine, consistently colocalizes with the amyloid deposits characteristic of Alzheimer's disease and may contribute to the generation of amyloid proteins and/or physically affect fibril assembly. In order to identify the structural domains of the amyloid-beta-peptide (Abeta) involved in the aggregation induced by AChE, we have studied the effect of this cholinergic enzyme on Abeta peptide fragments of different sizes. AChE enhanced the aggregation of the Abeta(12-28) and Abeta(25-35) peptides but not of the Abeta(1-16) fragment. The inductive effect of AChE on the aggregation of Abeta(12-28) was abolished by the presence of either Abeta(1-16) or Abeta(9-21). The effect of the enzyme was also analysed using two different mutant fragments, possessing a low and the other a high capacity for fibrillogenesis. The fragments used were Abeta(12-28)Val18-->Ala and Abeta(12-28)Glu22-->Gln, respectively. AChE was able to promote the aggregation of these fragments in a very specific way and both mutant peptides were able to form amyloid fibrils, as revealed by negative staining under the electron microscope. Binding assays indicated that AChE was bound to Abeta(12-28), as well as to the Abeta(1-16) peptide. AChE was seen to form strong complexes with the Abeta(12-28) fibrils as such complexes stained positively for both thioflavine-T and AChE activity, were resistant to high ionic strength treatment, and were partially sensitive to detergents, suggesting that hydrophobic interactions may play a role in the stabilization of the AChE-Abeta complex. Our results suggest that such amyloid-AChE complexes are formed when AChE interacts with the growing amyloid fibrils and accelerates the assembly of Abeta peptides. This is consistent with the fact that AChE is known to be present within Abeta deposits including the pre-amyloid diffuse and mature senile plaques found in Alzheimer's brain.
ESTHER : Alvarez_1997_J.Mol.Biol_272_348
PubMedSearch : Alvarez_1997_J.Mol.Biol_272_348
PubMedID: 9325095

Title : Acetylcholinesterase accelerates assembly of amyloid-beta-peptides into Alzheimer's fibrils: possible role of the peripheral site of the enzyme - Inestrosa_1996_Neuron_16_881
Author(s) : Inestrosa NC , Alvarez A , Perez CA , Moreno RD , Vicente M , Linker C , Casanueva OI , Soto C , Garrido J
Ref : Neuron , 16 :881 , 1996
Abstract : Acetylcholinesterase (AChE), an important component of cholinergic synapses, colocalizes with amyloid-beta peptide (A beta) deposits of Alzheimer's brain. We report here that bovine brain AChE, as well as the human and mouse recombinant enzyme, accelerates amyloid formation from wild-type A beta and a mutant A beta peptide, which alone produces few amyloid-like fibrils. The action of AChE was independent of the subunit array of the enzyme, was not affected by edrophonium, an active site inhibitor, but it was affected by propidium, a peripheral anionic binding site ligand. Butyrylcholinesterase, an enzyme that lacks the peripheral site, did not affect amyloid formation. Furthermore, AChE is a potent amyloid-promoting factor when compared with other A beta-associated proteins. Thus, in addition to its role in cholinergic synapses, AChE may function by accelerating A beta formation and could play a role during amyloid deposition in Alzheimer's brain.
ESTHER : Inestrosa_1996_Neuron_16_881
PubMedSearch : Inestrosa_1996_Neuron_16_881
PubMedID: 8608006

Title : Acetylcholinesterase, a senile plaque component, affects the fibrillogenesis of amyloid-beta-peptides - Alvarez_1995_Neurosci.Lett_201_49
Author(s) : Alvarez A , Bronfman F , Perez CA , Vicente M , Garrido J , Inestrosa NC
Ref : Neuroscience Letters , 201 :49 , 1995
Abstract : Acetylcholinesterase (AChE) colocalizes with amyloid-beta peptide (A beta) deposits present in the brain of Alzheimer's patients. Recent studies showed that A beta 1-40 can adopt two different conformational states in solution (an amyloidogenic conformer, A beta ac, and a non-amyloidogenic conformer, A beta nac) which have distinct abilities to form amyloid fibrils. We report here that AChE binds A beta nac and accelerates amyloid formation by the same peptide. No such effect was observed with A beta ac, the amyloidogenic conformer, suggesting that AChE acts as a 'pathological chaperone' inducing a conformational transition from A beta nac into A beta ac in vitro.
ESTHER : Alvarez_1995_Neurosci.Lett_201_49
PubMedSearch : Alvarez_1995_Neurosci.Lett_201_49
PubMedID: 96274631

Title : Anchorage of collagen-tailed acetylcholinesterase to the extracellular matrix is mediated by heparan sulfate proteoglycans - Brandan_1985_J.Cell.Biol_101_985
Author(s) : Brandan E , Maldonado M , Garrido J , Inestrosa NC
Ref : Journal of Cell Biology , 101 :985 , 1985
Abstract : Heparan sulfate and heparin, two sulfated glycosaminoglycans (GAGs), extracted collagen-tailed acetylcholinesterase (AChE) from the extracellular matrix (ECM) of the electric organ of Discopyge tschudii. The effect of heparan sulfate and heparin was abolished by protamine; other GAGs could not extract the esterase. The solubilization of the asymmetric AChE apparently occurs through the formation of a soluble AChE-GAG complex of 30S. Heparitinase treatment but not chondroitinase ABC treatment of the ECM released asymmetric AChE forms. This provides direct evidence for the vivo interaction between asymmetric AChE and heparan sulfate residues of the ECM. Biochemical analysis of the electric organ ECM showed that sulfated GAGs bound to proteoglycans account for 5% of the total basal lamina. Approximately 20% of the total GAGs were susceptible to heparitinase or nitrous acid oxidation which degrades specifically heparan sulfates, and approximately 80% were susceptible to digestion with chondroitinase ABC, which degrades chondroitin-4 and -6 sulfates and dermatan sulfate. Our experiments provide evidence that asymmetric AChE and carbohydrate components of proteoglycans are associated in the ECM; they also indicate that a heparan sulfate proteoglycan is involved in the anchorage of the collagen-tailed AChE to the synaptic basal lamina.
ESTHER : Brandan_1985_J.Cell.Biol_101_985
PubMedSearch : Brandan_1985_J.Cell.Biol_101_985
PubMedID: 3161900

Title : The electric organ of Discopyge tschudii: its innervated face and the biology of acetylcholinesterase - Mendez_1984_Cell.Mol.Neurobiol_4_125
Author(s) : Mendez B , Garrido J , Maldonado M , Jaksic FM , Inestrosa NC
Ref : Cellular Molecular Neurobiology , 4 :125 , 1984
Abstract : An ultrastructural, histochemical, and biochemical study of the electric organ of the South American Torpedinid ray, Discopyge tschudii, was carried out. Fine structural cytochemical localization of acetylcholinesterase (AChE) indicated that most of the esterase was associated with the basal lamina. Electron microscopy indicated no marked differences in the electrocyte ultrastructure between Discopyge and Torpedo californica. Discopyge electric organ possessed three molecular forms, two asymmetric forms (16 S and 13 S) and one globular hydrophobic form (6.5 S). The asymmetric 16 S AChE form was solubilized by heparin, a sulfated glycosaminoglycan, suggesting that heparin-like macromolecules are involved in the binding of the enzyme to the basal lamina. Our results show that cell-free translated AChE peptides, synthesized using Discopyge electric organ poly(A+) RNA, correspond to a main band of 62,000 daltons which probably represents the catalytic subunit of the asymmetric AChE.
ESTHER : Mendez_1984_Cell.Mol.Neurobiol_4_125
PubMedSearch : Mendez_1984_Cell.Mol.Neurobiol_4_125
PubMedID: 6488242