RÉSUMÉ
N-(Benzothiazole-2-yl)pyrrolamide DNA gyrase inhibitors with benzyl or phenethyl substituents attached to position 3 of the benzothiazole ring or to the carboxamide nitrogen atom were prepared and studied for their inhibition of Escherichia coli DNA gyrase by supercoiling assay. Compared to inhibitors bearing the substituents at position 4 of the benzothiazole ring, the inhibition was attenuated by moving the substituent to position 3 and further to the carboxamide nitrogen atom. A co-crystal structure of (Z)-3-benzyl-2-((4,5-dibromo-1H-pyrrole-2-carbonyl)imino)-2,3-dihydrobenzo[d]-thiazole-6-carboxylic acid (I) in complex with E. coli GyrB24 (ATPase subdomain) was solved, revealing the binding mode of this type of inhibitor to the ATP-binding pocket of the E. coli GyrB subunit. The key binding interactions were identified and their contribution to binding was rationalised by quantum theory of atoms in molecules (QTAIM) analysis. Our study shows that the benzyl or phenethyl substituents bound to the benzothiazole core interact with the lipophilic floor of the active site, which consists mainly of residues Gly101, Gly102, Lys103 and Ser108. Compounds with substituents at position 3 of the benzothiazole core were up to two orders of magnitude more effective than compounds with substituents at the carboxamide nitrogen. In addition, the 6-oxalylamino compounds were more potent inhibitors of E. coli DNA gyrase than the corresponding 6-acetamido analogues.
Sujet(s)
DNA gyrase , Escherichia coli , Inhibiteurs de la topoisomérase-II , Inhibiteurs de la topoisomérase-II/pharmacologie , Inhibiteurs de la topoisomérase-II/composition chimique , Inhibiteurs de la topoisomérase-II/synthèse chimique , DNA gyrase/métabolisme , DNA gyrase/composition chimique , Sites de fixation , Escherichia coli/enzymologie , Escherichia coli/effets des médicaments et des substances chimiques , Relation structure-activité , Benzothiazoles/composition chimique , Benzothiazoles/pharmacologie , Benzothiazoles/synthèse chimique , Adénosine triphosphate/métabolisme , Adénosine triphosphate/composition chimique , Structure moléculaire , Théorie quantique , Antibactériens/pharmacologie , Antibactériens/composition chimique , Antibactériens/synthèse chimique , Modèles moléculairesRÉSUMÉ
Multifunctional mitochondrial enzyme 17ß-hydroxysteroid dehydrogenase type 10 (17ß-HSD10) is a potential drug target for the treatment of various pathologies. The most discussed is the pathology associated with Alzheimer's disease (AD), where 17ß-HSD10 overexpression and its interaction with amyloid-ß peptide contribute to mitochondrial dysfunction and neuronal stress. In this work, a series of new benzothiazole-derived 17ß-HSD10 inhibitors were designed based on the structure-activity relationship analysis of formerly published inhibitors. A set of enzyme-based and cell-based methods were used to evaluate the inhibitory potency of new compounds, their interaction with the enzyme, and their cytotoxicity. Most compounds exhibited significantly a higher inhibitory potential compared to published benzothiazolyl ureas and good target engagement in a cellular environment accompanied by low cytotoxicity. The best hits displayed mixed-type inhibition with half maximal inhibitory concentration (IC50) values in the nanomolar range for the purified enzyme (3-7, 15) and/or low micromolar IC50 values in the cell-based assay (6, 13-16).
RÉSUMÉ
Mitochondrial enzyme 17ß-hydroxysteroid dehydrogenase type 10 (HSD10) is a potential molecular target for treatment of mitochondrial-related disorders such as Alzheimer's disease (AD). Its over-expression in AD brains is one of the critical factors disturbing the homeostasis of neuroprotective steroids and exacerbating amyloid beta (Aß)-mediated mitochondrial toxicity and neuronal stress. This study was focused on revalidation of the most potent HSD10 inhibitors derived from benzothiazolyl urea scaffold using fluorescent-based enzymatic assay with physiologically relevant substrates of 17ß-oestradiol and allopregnanolone. The oestradiol-based assay led to the identification of two nanomolar inhibitors (IC50 70 and 346 nM) differing from HSD10 hits revealed from the formerly used assay. Both identified inhibitors were found to be effective also in allopregnanolone-based assay with non-competitive or uncompetitive mode of action. In addition, both inhibitors were confirmed to penetrate the HEK293 cells and they were able to inhibit the HSD10 enzyme in the cellular environment. Both molecules seem to be potential lead structures for further research and development of HDS10 inhibitors.
RÉSUMÉ
17ß-hydroxysteroid dehydrogenase type 10 (17ß-HSD10) is a multifunctional mitochondrial enzyme and putative drug target for the treatment of various pathologies including Alzheimer's disease or some types of hormone-dependent cancer. In this study, a series of new benzothiazolylurea-based inhibitors were developed based on the structure-activity relationship (SAR) study of previously published compounds and predictions of their physico-chemical properties. This led to the identification of several submicromolar inhibitors (IC50 â¼0.3 µM), the most potent compounds within the benzothiazolylurea class known to date. The positive interaction with 17ß-HSD10 was further confirmed by differential scanning fluorimetry and the best molecules were found to be cell penetrable. In addition, the best compounds weren't found to have additional effects for mitochondrial off-targets and cytotoxic or neurotoxic effects. The two most potent inhibitors 9 and 11 were selected for in vivo pharmacokinetic study after intravenous and peroral administration. Although the pharmacokinetic results were not fully conclusive, it seemed that compound 9 was bioavailable after peroral administration and could penetrate into the brain (brain-plasma ratio 0.56).
Sujet(s)
Maladie d'Alzheimer , Humains , Maladie d'Alzheimer/traitement médicamenteux , Relation structure-activité , 17-Hydroxysteroid dehydrogenases , Encéphale/métabolisme , Antienzymes/composition chimiqueRÉSUMÉ
Cyclophilin D (CypD) is a key regulator of mitochondrial permeability transition pore (mPTP) opening. This pathophysiological phenomenon is associated with the development of several human diseases, including ischemia-reperfusion injury and neurodegeneration. Blocking mPTP opening through CypD inhibition could be a novel and promising therapeutic approach for these conditions. While numerous CypD inhibitors have been discovered to date, none have been introduced into clinical practice, mostly owing to their high toxicity, unfavorable pharmacokinetics, and low selectivity for CypD over other cyclophilins. This review summarizes current knowledge of CypD inhibitors, with a particular focus on small-molecule compounds with regard to their in vitro activity, their selectivity for CypD, and their binding mode within the enzyme's active site. Finally, approaches for improving the molecular design of CypD inhibitors are discussed.
Sujet(s)
Maladies mitochondriales , Protéines de transport de la membrane mitochondriale , Peptidyl-prolyl isomerase F , Peptidyl-prolyl isomerase F/antagonistes et inhibiteurs , Humains , Mitochondries/métabolisme , Maladies mitochondriales/traitement médicamenteux , Protéines de transport de la membrane mitochondriale/métabolisme , Pore de transition de perméabilité mitochondrialeRÉSUMÉ
The rise in multidrug-resistant bacteria defines the need for identification of new antibacterial agents that are less prone to resistance acquisition. Compounds that simultaneously inhibit multiple bacterial targets are more likely to suppress the evolution of target-based resistance than monotargeting compounds. The structurally similar ATP binding sites of DNA gyrase and topoisomerase â £ offer an opportunity to accomplish this goal. Here we present the design and structure-activity relationship analysis of balanced, low nanomolar inhibitors of bacterial DNA gyrase and topoisomerase IV that show potent antibacterial activities against the ESKAPE pathogens. For inhibitor 31c, a crystal structure in complex with Staphylococcus aureus DNA gyrase B was obtained that confirms the mode of action of these compounds. The best inhibitor, 31h, does not show any in vitro cytotoxicity and has excellent potency against Gram-positive (MICs: range, 0.0078-0.0625 µg/mL) and Gram-negative pathogens (MICs: range, 1-2 µg/mL). Furthermore, 31h inhibits GyrB mutants that can develop resistance to other drugs. Based on these data, we expect that structural derivatives of 31h will represent a step toward clinically efficacious multitargeting antimicrobials that are not impacted by existing antimicrobial resistance.
Sujet(s)
Adénosine triphosphate/pharmacologie , Antibactériens/pharmacologie , DNA gyrase/métabolisme , DNA topoisomerase IV/antagonistes et inhibiteurs , Escherichia coli/effets des médicaments et des substances chimiques , Staphylococcus aureus/effets des médicaments et des substances chimiques , Adénosine triphosphate/synthèse chimique , Adénosine triphosphate/composition chimique , Antibactériens/synthèse chimique , Antibactériens/composition chimique , Cristallographie aux rayons X , DNA topoisomerase IV/métabolisme , Relation dose-effet des médicaments , Escherichia coli/enzymologie , Escherichia coli/pathogénicité , Tests de sensibilité microbienne , Simulation de docking moléculaire , Structure moléculaire , Staphylococcus aureus/enzymologie , Staphylococcus aureus/pathogénicité , Relation structure-activitéRÉSUMÉ
Mitochondrial enzymes are targets of newly synthesized drugs being tested for the treatment of neurodegenerative disorders, such as Alzheimer's disease (AD). The enzyme 17ß-hydroxysteroid dehydrogenase type 10 (HSD10) is a multifunctional mitochondrial protein that is thought to play a role in the pathophysiology of AD and is one of the targets of new potential AD drugs. The in vitro effects of frentizole, riluzole, AG18051, and 42 novel modulators of HSD10 (potential AD drugs) on citrate synthase (CS) activity, monoamine oxidase (MAO) activity, complex I- or complex II-linked mitochondrial respiratory rate, and complex I activity were measured in isolated pig brain mitochondria. Based on their minimal inhibitory effects on the respiratory rate of mitochondria and CS and complex I activity, six novel compounds were selected for further testing. Assuming that inhibition of MAO-B could be a desirable effect of AD drugs, only AG18051 and one new compound met the criteria for MAO-B inhibition with minimal drug-induced effects on mitochondrial respiration. In conclusion, our in vitro screening of mitochondrial effect of novel potential AD drugs has enabled the selection of the most promising molecules for further testing that are relatively safe in terms of drug-induced mitochondrial toxicity.
Sujet(s)
17-Hydroxysteroid dehydrogenases/antagonistes et inhibiteurs , 17-Hydroxysteroid dehydrogenases/toxicité , Respiration cellulaire/effets des médicaments et des substances chimiques , Antienzymes/toxicité , Antienzymes/usage thérapeutique , Mitochondries/effets des médicaments et des substances chimiques , Maladies neurodégénératives/traitement médicamenteux , Animaux , Humains , Modèles animaux , SuidaeRÉSUMÉ
We designed and synthesized a series of inhibitors of the bacterial enzymes DNA gyrase and DNA topoisomerase IV, based on our recently published benzothiazole-based inhibitor bearing an oxalyl moiety. To improve the antibacterial activity and retain potent enzymatic activity, we systematically explored the chemical space. Several strategies of modification were followed: varying substituents on the pyrrole carboxamide moiety, alteration of the central scaffold, including variation of substitution position and, most importantly, modification of the oxalyl moiety. Compounds with acidic, basic, and neutral properties were synthesized. To understand the mechanism of action and binding mode, we have obtained a crystal structure of compound 16a, bearing a primary amino group, in complex with the N-terminal domain of E. coli gyrase B (24 kDa) (PDB: 6YD9). Compound 15a, with a low molecular weight of 383 Da, potent inhibitory activity on E. coli gyrase (IC50 = 9.5 nM), potent antibacterial activity on E. faecalis (MIC = 3.13 µM), and efflux impaired E. coli strain (MIC = 0.78 µM), is an important contribution for the development of novel gyrase and topoisomerase IV inhibitors in Gram-negative bacteria.
RÉSUMÉ
Berberine, a naturally occurring compound, possesses an interesting multipotent pharmacological profile potentially applicable for Alzheimer's disease (AD) treatment. In this study, a series of novel 22 berberine derivatives was developed and tested in vitro. Berberine core was substituted at position 9-O of its aromatic ring region. All the hybrids under the study revealed multi-targeted profile inhibiting prolyl oligopeptidase, acetylcholinesterase and butyrylcholinesterase highlighting 4a, 4g, 4j, 4l and 4s possessing balanced activities in the micromolar range. The top-ranked candidates in terms of the most pronounced potency against POP, AChE and BChE can be classified as 4d, 4u and 4v, bearing 4-methylbenzyl, (naphthalen-2-yl)methylene and 1-phenoxyethyl moieties, respectively. In vitro data were corroborated by detailed kinetic analysis of the selected lead molecules. 4d, 4u and 4v were also inspected for their potential to inhibit aggregation of two abberant proteins in AD, namely amyloid beta and tau, indicating their potential disease-modifying properties. To explain the results of our study, we carried out docking simulation to the active sites of the respective enzyme with the best berberine derivatives, along with QSAR study. We also investigated compounds' potential permeability through blood-brain barrier by applying parallel artificial membrane permeation assay and addressed their cytotoxicity profile.
Sujet(s)
Berbérine/composition chimique , Berbérine/pharmacologie , Anticholinestérasiques/composition chimique , Anticholinestérasiques/pharmacologie , Cholinesterases/métabolisme , Conception de médicament , Prolyl-oligopeptidases/antagonistes et inhibiteurs , Berbérine/métabolisme , Barrière hémato-encéphalique/métabolisme , Lignée cellulaire tumorale , Anticholinestérasiques/métabolisme , HumainsRÉSUMÉ
Alzheimer's disease (AD) has a complex pathophysiology that includes aggregation of pathological proteins, impaired neurotransmission, increased oxidative stress, or microglia-mediated neuroinflammation. Therapeutics targeting only one of these AD-related subpathologies have not yet been successful in the search for a disease-modifying treatment. Therefore, multi-target drugs (MTDs) aiming simultaneously at several subpathologies are expected to be a better approach. However, the concept of MTD is inherently connected with several limitations, which are often ignored during MTD design and development. Here, we provide an overview of the MTD approach and discuss its potential pitfalls in the context of AD treatment. We also put forward ideas to be used in the rational design of MTDs to obtain drugs that are effective against AD.
Sujet(s)
Maladie d'Alzheimer , Préparations pharmaceutiques , Maladie d'Alzheimer/traitement médicamenteux , Systèmes de délivrance de médicaments , Humains , Inflammation , Stress oxydatif , Transmission synaptiqueRÉSUMÉ
17ß-hydroxysteroid dehydrogenase (17ß-HSD10) is a multifunctional human enzyme with important roles both as a structural component and also as a catalyst of many metabolic pathways. This mitochondrial enzyme has important functions in the metabolism, development and aging of the neural system, where it is involved in the homeostasis of neurosteroids, especially in regard to estradiol, changes in which make it an essential part of neurodegenerative pathology. These roles therefore, indicate that 17ß-HSD10 may be a possible druggable target for neurodegenerative diseases including Alzheimer's disease (AD), and in hormone-dependent cancer. The objective of this review was to provide a summary about physiological functions and pathological roles of 17ß-HSD10 and the modulators of its activity.
Sujet(s)
3-Hydroxyacyl-CoA dehydrogenases/génétique , 3-Hydroxyacyl-CoA dehydrogenases/métabolisme , État de santé , Mitochondries/métabolisme , 3-Hydroxyacyl-CoA dehydrogenases/composition chimique , Maladie d'Alzheimer/génétique , Maladie d'Alzheimer/métabolisme , Humains , Mitochondries/génétique , Mutation/génétique , Tumeurs/génétique , Tumeurs/métabolisme , Maladies neurodégénératives/génétique , Maladies neurodégénératives/métabolisme , Structure secondaire des protéinesRÉSUMÉ
Human cyclophilin D is a mitochondrial peptidyl-prolyl isomerase that plays a role in regulating the opening of the mitochondrial permeability transition pore. It is considered a viable and promising molecular target for the treatment of diseases for which disease development is associated with pore opening, e.g., Alzheimer's disease or ischemia/reperfusion injury. Currently available and widely used in vitro methods based on Kofron's assay for determining cyclophilin D activity suffer from serious drawbacks and limitations. In this study, a completely novel approach for an in vitro assay of cyclophilin D activity using RNase T1 refolding is introduced. The method is simple and is more in line with the presumed physiological role of cyclophilin D in protein folding than Kofron's assay, which relies on a peptide substrate. The method is applicable for identifying novel inhibitors of cyclophilin D as potential drugs for the treatment of the diseases mentioned above. Moreover, the description of CypD activity in the in vitro RNase T1 refolding assay reveals new possibilities for investigating the role of cyclophilin D in protein folding in cells and may lead to a better understanding of its pathological and physiological roles.
Sujet(s)
Découverte de médicament , Mitochondries/métabolisme , Peptidyl-prolyl isomerase F/métabolisme , Repliement des protéines , Ribonuclease T1/composition chimique , Animaux , Aspergillus oryzae/enzymologie , Bovins , Peptidyl-prolyl isomerase F/composition chimique , Humains , Modèles moléculaires , Conformation des protéinesRÉSUMÉ
Human 17ß-hydroxysteroid dehydrogenase type 10 is a multifunctional protein involved in many enzymatic and structural processes within mitochondria. This enzyme was suggested to be involved in several neurological diseases, e.g., mental retardation, Parkinson's disease, or Alzheimer's disease, in which it was shown to interact with the amyloid-beta peptide. We prepared approximately 60 new compounds based on a benzothiazolyl scaffold and evaluated their inhibitory ability and mechanism of action. The most potent inhibitors contained 3-chloro and 4-hydroxy substitution on the phenyl ring moiety, a small substituent at position 6 on the benzothiazole moiety, and the two moieties were connected via a urea linker (4at, 4bb, and 4bg). These compounds exhibited IC50 values of 1-2 µM and showed an uncompetitive mechanism of action with respect to the substrate, acetoacetyl-CoA. These uncompetitive benzothiazolyl inhibitors of 17ß-hydroxysteroid dehydrogenase type 10 are promising compounds for potential drugs for neurodegenerative diseases that warrant further research and development.
Sujet(s)
3-Hydroxyacyl-CoA dehydrogenases/antagonistes et inhibiteurs , Benzothiazoles/composition chimique , Antienzymes/composition chimique , Antienzymes/pharmacologie , Urée/composition chimique , Urée/pharmacologie , 3-Hydroxyacyl-CoA dehydrogenases/composition chimique , Maladie d'Alzheimer/traitement médicamenteux , Activation enzymatique , Humains , Cinétique , Structure moléculaire , Protéines recombinantes , Relation structure-activitéRÉSUMÉ
: It has long been established that mitochondrial dysfunction in Alzheimer's disease (AD) patients can trigger pathological changes in cell metabolism by altering metabolic enzymes such as the mitochondrial 17ß-hydroxysteroid dehydrogenase type 10 (17ß-HSD10), also known as amyloid-binding alcohol dehydrogenase (ABAD). We and others have shown that frentizole and riluzole derivatives can inhibit 17ß-HSD10 and that this inhibition is beneficial and holds therapeutic merit for the treatment of AD. Here we evaluate several novel series based on benzothiazolylurea scaffold evaluating key structural and activity relationships required for the inhibition of 17ß-HSD10. Results show that the most promising of these compounds have markedly increased potency on our previously published inhibitors, with the most promising exhibiting advantageous features like low cytotoxicity and target engagement in living cells.
Sujet(s)
17-Hydroxysteroid dehydrogenases/antagonistes et inhibiteurs , 17-Hydroxysteroid dehydrogenases/composition chimique , Benzothiazoles/composition chimique , Urée/composition chimique , Maladie d'Alzheimer/traitement médicamenteux , Peptides bêta-amyloïdes/métabolisme , Lignée cellulaire , Relation dose-effet des médicaments , Conception de médicament , Humains , Mitochondries/métabolisme , Structure moléculaire , Relation structure-activitéRÉSUMÉ
Alzheimer's Disease (AD) is a multifactorial progressive neurodegenerative disorder characterized by memory loss, disorientation, and gradual deterioration of intellectual capacity. Its etiology has not been elucidated yet. To date, only one therapeutic approach has been approved for the treatment of AD. The pharmacotherapy of AD has relied on noncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist - memantine, and acetylcholinesterase (AChE) inhibitors (AChEIs) - tacrine, donepezil, rivastigmine and galantamine. Donepezil was able to ameliorate the symptoms related to AD mainly via AChE, but also through reduction of ß-amyloid burden. This review presents the overview of donepezilrelated compounds as potential anti-AD drugs developed on the basis of cholinergic hypothesis to act as solely AChE and butyrylcholinesterase (BChE) inhibitors.
Sujet(s)
Maladie d'Alzheimer/traitement médicamenteux , Anticholinestérasiques/pharmacologie , Anticholinestérasiques/usage thérapeutique , Donépézil/pharmacologie , Donépézil/usage thérapeutique , Neuroprotecteurs/pharmacologie , Neuroprotecteurs/usage thérapeutique , Acetylcholinesterase/métabolisme , Maladie d'Alzheimer/métabolisme , Animaux , Butyrylcholine esterase/métabolisme , Anticholinestérasiques/composition chimique , Donépézil/composition chimique , Humains , Structure moléculaire , Neuroprotecteurs/composition chimiqueRÉSUMÉ
Several neurodegenerative disorders including Alzheimer's disease (AD) have been connected with deregulation of casein kinase 1 (CK1) activity. Inhibition of CK1 therefore presents a potential therapeutic strategy against such pathologies. Recently, novel class of CK1-specific inhibitors with N-(benzo[d]thiazol-2-yl)-2-phenylacetamide structural scaffold has been discovered. 1-(benzo[d]thiazol-2-yl)-3-phenylureas, on the other hand, are known inhibitors amyloid-beta binding alcohol dehydrogenase (ABAD), an enzyme also involved in pathophysiology of AD. Based on their tight structural similarity, we decided to evaluate series of previously published benzothiazolylphenylureas, originally designed as ABAD inhibitors, for their inhibitory activity towards CK1. Several compounds were found to be submicromolar CK1 inhibitors. Moreover, two compounds were found to inhibit both, ABAD and CK1. Such dual-activity could be of advantage for AD treatment, as it would simultaneously target two distinct pathological processes involved in disease's progression. Based on PAMPA testing both compounds were suggested to permeate the blood-brain barrier, which makes them, together with their unique dual activity, interesting lead compounds for further development.
Sujet(s)
3-Hydroxyacyl-CoA dehydrogenases/métabolisme , Casein kinase I/antagonistes et inhibiteurs , Antienzymes/pharmacologie , Maladies neurodégénératives/traitement médicamenteux , Phénylurées/pharmacologie , Casein kinase I/métabolisme , Relation dose-effet des médicaments , Antienzymes/composition chimique , Humains , Structure moléculaire , Maladies neurodégénératives/métabolisme , Phénylurées/composition chimique , Relation structure-activitéRÉSUMÉ
The purpose of this study was to identify new small molecules that possess activity on human toll-like receptor 4 associated with the myeloid differentiation protein 2 (hTLR4/MD2). Following current rational drug design principles, we firstly performed a ligand and structure based virtual screening of more than 130â¯000 compounds to discover until now unknown class of hTLR4/MD2 modulators that could be used as novel type of immunologic adjuvants. The core of the in silico study was molecular docking of flexible ligands in a partially flexible hTLR4/MD2 receptor model using a peta-flops-scale supercomputer. The most promising substances resulting from this study, related to anthracene-succimide hybrids, were synthesized and tested. The best prepared candidate exhibited 80% of Monophosphoryl Lipid A in vitro agonistic activity in cell lines expressing hTLR4/MD2.
Sujet(s)
Simulation numérique , Conception de médicament , Bibliothèques de petites molécules/pharmacologie , Récepteur de type Toll-4/antagonistes et inhibiteurs , Lignée cellulaire , Relation dose-effet des médicaments , Évaluation préclinique de médicament , Humains , Ligands , Simulation de docking moléculaire , Structure moléculaire , Bibliothèques de petites molécules/synthèse chimique , Bibliothèques de petites molécules/composition chimique , Relation structure-activitéRÉSUMÉ
Alzheimer's disease (AD) is a neurodegenerative disorder associated with an excessive accumulation of amyloid-beta peptide (Aß). Based on the multifactorial nature of AD, preparation of multi-target-directed ligands presents a viable option to address more pathological events at one time. A novel class of asymmetrical disubstituted indolyl thioureas have been designed and synthesized to interact with monoamine oxidase (MAO) and/or amyloid-binding alcohol dehydrogenase (ABAD). The design combines the features of known MAO inhibitors scaffolds (e.g. rasagiline or ladostigil) and a frentizole moiety with potential to interact with ABAD. Evaluation against MAO identified several compounds that inhibited in the low to moderate micromolar range. The most promising compound (19) inhibited human MAO-A and MAO-B with IC50 values of 6.34µM and 0.30µM, respectively. ABAD activity evaluation did not show any highly potent compound, but the compound series allowed identification of structural features to assist the future development of ABAD inhibitors. Finally, several of the compounds were found to be potent inhibitors of horseradish peroxidase (HRP), preventing the use of the Amplex™ Red assay to detect hydrogen peroxide produced by MAO, highlighting the need for serious precautions when using an enzyme-coupled assay.
Sujet(s)
3-Hydroxyacyl-CoA dehydrogenases/antagonistes et inhibiteurs , Maladie d'Alzheimer/traitement médicamenteux , Benzothiazoles/pharmacologie , Antienzymes/pharmacologie , Monoamine oxidase/métabolisme , Phénylurées/pharmacologie , Thiourée/pharmacologie , 3-Hydroxyacyl-CoA dehydrogenases/métabolisme , Maladie d'Alzheimer/métabolisme , Benzothiazoles/composition chimique , Relation dose-effet des médicaments , Antienzymes/synthèse chimique , Antienzymes/composition chimique , Humains , Structure moléculaire , Phénylurées/composition chimique , Relation structure-activité , Thiourée/synthèse chimique , Thiourée/composition chimiqueRÉSUMÉ
BACKGROUND: The mitochondrial enzyme amyloid beta-binding alcohol dehydrogenase (ABAD) also known as 17ß-hydroxysteroid dehydrogenase type 10 (17ß-HSD10) has been connected with the pathogenesis of Alzheimer's disease (AD). ABAD/ 17ß-HSD10 is a binding site for the amyloid-beta peptide (Aß) inside the mitochondrial matrix where it exacerbates Aß toxicity. Interaction between these two proteins triggers a series of events leading to mitochondrial dysfunction as seen in AD. METHODS: As ABAD's enzymatic activity is required for mediating Aß toxicity, its inhibition presents a promising strategy for AD treatment. In this study, a series of new benzothiazolylurea analogues have been prepared and evaluated in vitro for their potency to inhibit ABAD/ 17ß-HSD10 enzymatic activity. The most potent compounds have also been tested for their cytotoxic properties and their ability to permeate through blood-brain barrier has been predicted. To explain the structure-activity relationship QSAR and pharmacophore studies have been performed. RESULTS AND CONCLUSIONS: Compound 12 was identified being the most promising hit compound with good inhibitory activity (IC50 = 3.06 ± 0.40µM) and acceptable cytotoxicity profile comparable to the parent compound of frentizole. The satisfactory physical-chemical properties suggesting its capability to permeate through BBB make compound 12 a novel lead structure for further development and biological assessment.
RÉSUMÉ
Amyloid-beta peptide (Aß) has been recognized to interact with numerous proteins, which may lead to pathological changes in cell metabolism of Alzheimer's disease (AD) patients. One such known metabolic enzyme is mitochondrial amyloid-binding alcohol dehydrogenase (ABAD), also known as 17ß-hydroxysteroid dehydrogenase type 10 (17ß-HSD10). Altered enzyme function caused by the Aß-ABAD interaction, was previously shown to cause mitochondrial distress and a consequent cytotoxic effect, therefore providing a feasible target in AD drug development. Based on previous frentizole derivatives studies, we report two novel series of benzothiazolyl ureas along with novel insights into the structure and activity relationships for inhibition of ABAD. Two compounds (37, 39) were identified as potent ABAD inhibitors, where compound 39 exhibited comparable cytotoxicity with the frentizole standard; however, one-fold higher cytotoxicity than the parent riluzole standard. The calculated and experimental physical chemical properties of the most potent compounds showed promising features for blood-brain barrier penetration.