RESUMO
AIMS: KCL-286 is an orally available agonist that activates the retinoic acid receptor (RAR) ß2, a transcription factor which stimulates axonal outgrowth. The investigational medicinal product is being developed for treatment of spinal cord injury (SCI). This adaptive dose escalation study evaluated the tolerability, safety and pharmacokinetics and pharmacodynamic activity of KCL-286 in male healthy volunteers to establish dosing to be used in the SCI patient population. METHODS: The design was a double blind, randomized, placebo-controlled dose escalation study in 2 parts: a single ascending dose adaptive design with a food interaction arm, and a multiple ascending dose design. RARß2 mRNA expression was evaluated in white blood cells. RESULTS: At the highest single and multiple ascending doses (100 mg), no trends or clinically important differences were noted in the incidence or intensity of adverse events (AEs), serious AEs or other safety assessments with none leading to withdrawal from the study. The AEs were dry skin, rash, skin exfoliation, raised liver enzymes and eye disorders. There was an increase in mean maximum observed concentration and area under the plasma concentration-time curve up to 24 h showing a trend to subproportionality with dose. RARß2 was upregulated by the investigational medicinal product in white blood cells. CONCLUSION: KCL-286 was well tolerated by healthy human participants following doses that exceeded potentially clinically relevant plasma exposures based on preclinical in vivo models. Target engagement shows the drug candidate activates its receptor. These findings support further development of KCL-286 as a novel oral treatment for SCI.
Assuntos
Drogas em Investigação , Receptores do Ácido Retinoico , Humanos , Masculino , Voluntários Saudáveis , Relação Dose-Resposta a Droga , Área Sob a Curva , Método Duplo-CegoRESUMO
In the CNS, oligodendrocytes are responsible for myelin formation and maintenance. Following spinal cord injury, oligodendrocyte loss and an inhibitory milieu compromise remyelination and recovery. Here, we explored the role of retinoic acid receptor-beta (RARß) signaling in remyelination. Using a male Sprague Dawley rat model of PNS-CNS injury, we show that oral treatment with a novel drug like RARß agonist, C286, induces neuronal expression of the proteoglycan decorin and promotes myelination and differentiation of oligodendrocyte precursor cells (NG2+ cells) in a decorin-mediated neuron-glia cross talk. Decorin promoted the activation of RARα in NG2+ cells by increasing the availability of the endogenous ligand RA. NG2+ cells synthesize RA, which is released in association with exosomes. We found that decorin prevents this secretion through regulation of the EGFR-calcium pathway. Using functional and pharmacological studies, we further show that RARα signaling is both required and sufficient for oligodendrocyte differentiation. These findings illustrate that RARß and RARα are important regulators of oligodendrocyte differentiation, providing new targets for myelination.SIGNIFICANCE STATEMENT This study identifies novel therapeutic targets for remyelination after PNS-CNS injury. Pharmacological and knock-down experiments show that the retinoic acid (RA) signaling promotes differentiation of oligodendrocyte precursor cells (OPCs) and remyelination in a cross talk between neuronal RA receptor-beta (RARß) and RARα in NG2+ cells. We show that stimulation of RARα is required for the differentiation of OPCs and we describe for the first time how oral treatment with a RARß agonist (C286, currently being tested in a Phase 1 trial, ISRCTN12424734) leads to the endogenous synthesis of RA through retinaldehyde dehydrogenase 2 (Raldh2) in NG2 cells and controls exosome-associated-RA intracellular levels through a decorin-Ca2+ pathway. Although RARß has been implicated in distinct aspects of CNS regeneration, this study identifies a novel function for both RARß and RARα in remyelination.
Assuntos
Exossomos/metabolismo , Bainha de Mielina/metabolismo , Regeneração Nervosa/efeitos dos fármacos , Receptores do Ácido Retinoico/agonistas , Traumatismos da Medula Espinal/tratamento farmacológico , Tretinoína/metabolismo , Animais , Decorina/metabolismo , Receptores ErbB/metabolismo , Bainha de Mielina/efeitos dos fármacos , Regeneração Nervosa/fisiologia , Oligodendroglia/efeitos dos fármacos , Oligodendroglia/metabolismo , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Traumatismos da Medula Espinal/metabolismoRESUMO
Oxadiazole replacement of an amide linkage in an RARα agonist template 1, followed by lead optimisation, has produced a highly potent and selective RARß agonist 4-(5-(4,7-dimethylbenzofuran-2-yl)-1,2,4-oxadiazol-3-yl)benzoic acid (10) with good oral bioavailability in the rat and dog. This molecule increases neurite outgrowth in vitro and induces sensory axon regrowth in vivo in a rodent model of avulsion and crush injury, and thus has the potential for the treatment of nerve injury.
Assuntos
Oxidiazóis/química , Receptores do Ácido Retinoico/agonistas , Administração Oral , Animais , Cães , Avaliação Pré-Clínica de Medicamentos , Meia-Vida , Locomoção/efeitos dos fármacos , Células Madin Darby de Rim Canino , Crescimento Neuronal/efeitos dos fármacos , Traumatismos do Nervo Óptico/tratamento farmacológico , Oxidiazóis/farmacocinética , Oxidiazóis/farmacologia , Ratos , Receptores do Ácido Retinoico/metabolismo , Relação Estrutura-AtividadeRESUMO
Stimulation of retinoic acid (RA) mediated signalling pathways following neural injury leads to regeneration in the adult nervous system and numerous studies have shown that the specific activation of the retinoic acid receptor ß (RARß) is required for this process. Here we identify a novel mechanism by which neuronal RARß activation results in the endogenous synthesis of RA which is released in association with exosomes and acts as a positive cue to axonal/neurite outgrowth. Using an established rodent model of RARß induced axonal regeneration, we show that neuronal RARß activation upregulates the enzymes involved in RA synthesis in a cell specific manner; alcohol dehydrogenase7 (ADH7) in neurons and aldehyde dehydrogenase 2 (Raldh2) in NG2 expressing cells (NG2+ cells). These release RA in association with exosomes providing a permissive substrate to neurite outgrowth. Conversely, deletion of Raldh2 in the NG2+ cells in our in vivo regeneration model is sufficient to compromise axonal outgrowth. This hitherto unidentified RA paracrine signalling is required for axonal/neurite outgrowth and is initiated by the activation of neuronal RARß signalling.
Assuntos
Antígenos/metabolismo , Exossomos/metabolismo , Regeneração Nervosa/fisiologia , Neuroglia/metabolismo , Crescimento Neuronal/fisiologia , Proteoglicanas/metabolismo , Tretinoína/metabolismo , Aldeído Oxirredutases/metabolismo , Animais , Transporte Biológico/fisiologia , Células Cultivadas , Medula Cervical/metabolismo , Medula Cervical/patologia , Técnicas de Cocultura , Modelos Animais de Doenças , Exossomos/patologia , Masculino , Camundongos , Neuroglia/patologia , Neurônios/metabolismo , Neurônios/patologia , Ratos Sprague-Dawley , Receptores do Ácido Retinoico/metabolismo , Retinal Desidrogenase/metabolismo , Raízes Nervosas Espinhais/lesões , Raízes Nervosas Espinhais/metabolismo , Raízes Nervosas Espinhais/patologiaRESUMO
A ligand-based virtual screening exercise examining likely bioactive conformations of AM 580 (2) and AGN 193836 (3) was used to identify the novel, less lipophilic RARα agonist 4-(3,5-dichloro-4-ethoxybenzamido)benzoic acid 5, which has good selectivity over the RARß, and RARγ receptors. Analysis of the medicinal chemistry parameters of the 3,5-substituents of derivatives of template 5 enabled us to design a class of drug-like molecules with lower intrinsic clearance and higher oral bioavailability which led to the novel RARα agonist 4-(3-chloro-4-ethoxy-5-isopropoxybenzamido)-2-methylbenzoic acid 56 that has high RARα potency and excellent selectivity versus RARß (2 orders of magnitude) and RARγ (4 orders of magnitude) at both the human and mouse RAR receptors with improved drug-like properties. This RARα specific agonist 56 has high oral bioavailability (>80%) in both mice and dogs with a good PK profile and was shown to be inactive in cytotoxicity and genotoxicity screens.
Assuntos
Aminobenzoatos/química , Benzoatos/química , Desenho de Fármacos , Receptor alfa de Ácido Retinoico/agonistas , Tetra-Hidronaftalenos/química , Administração Oral , Aminobenzoatos/farmacocinética , Aminobenzoatos/toxicidade , Animais , Benzoatos/farmacocinética , Benzoatos/toxicidade , Células COS , Sobrevivência Celular/efeitos dos fármacos , Chlorocebus aethiops , Meia-Vida , Células Hep G2 , Humanos , Camundongos , Microssomos Hepáticos/metabolismo , Ratos , Receptores do Ácido Retinoico/agonistas , Receptores do Ácido Retinoico/metabolismo , Receptor alfa de Ácido Retinoico/metabolismo , Relação Estrutura-Atividade , Tetra-Hidronaftalenos/farmacocinética , Tetra-Hidronaftalenos/toxicidade , Receptor gama de Ácido RetinoicoRESUMO
Failure of axonal regeneration in the central nervous system (CNS) is mainly attributed to a lack of intrinsic neuronal growth programs and an inhibitory environment from a glial scar. Phosphatase and tensin homolog (PTEN) is a major negative regulator of neuronal regeneration and, as such, inhibiting its activity has been considered a therapeutic target for spinal cord (SC) injuries (SCIs). Using a novel model of rat cervical avulsion, we show that treatment with a retinoic acid receptor ß (RARß) agonist results in locomotor and sensory recovery. Axonal regeneration from the severed roots into the SC could be seen by biotinylated dextran amine labeling. Light micrographs of the dorsal root entry zone show the peripheral nervous system (PNS)-CNS transition of regrown axons. RARß agonist treatment also resulted in the absence of scar formation. Mechanism studies revealed that, in RARß-agonist-treated neurons, PTEN activity is decreased by cytoplasmic phosphorylation and increased secretion in exosomes. These are taken up by astrocytes, resulting in hampered proliferation and causing them to arrange in a normal-appearing scaffold around the regenerating axons. Attribution of the glial modulation to neuronal PTEN in exosomes was demonstrated by the use of an exosome inhibitor in vivo and PTEN siRNA in vitro assays. The dual effect of RARß signaling, both neuronal and neuronal-glial, results in axonal regeneration into the SC after dorsal root neurotmesis. Targeting this pathway may open new avenues for the treatment of SCIs. SIGNIFICANCE STATEMENT: Spinal cord injuries (SCIs) often result in permanent damage in the adult due to the very limited capacity of axonal regeneration. Intrinsic neuronal programs and the formation of a glial scar are the main obstacles. Here, we identify a single target, neuronal retinoic acid receptor ß (RARß), which modulates these two aspects of the postinjury physiological response. Activation of RARß in the neuron inactivates phosphatase and tensin homolog and induces its transfer into the astrocytes in small vesicles, where it prevents scar formation. This may open new therapeutic avenues for SCIs.
Assuntos
Astrócitos/metabolismo , Cicatriz/metabolismo , Exossomos/metabolismo , Neuroglia/metabolismo , PTEN Fosfo-Hidrolase/metabolismo , Receptores do Ácido Retinoico/fisiologia , Regeneração da Medula Espinal/fisiologia , Animais , Células Cultivadas , Cicatriz/prevenção & controle , Masculino , Camundongos , Neuroglia/patologia , Neurônios/metabolismo , Ratos , Ratos Wistar , Transdução de Sinais/fisiologiaRESUMO
The retinoic acid receptor (RAR) α system plays a key role in the adult brain, participating in the homeostatic control of synaptic plasticity, essential for memory function. Here we show that RARα signalling is down-regulated by amyloid beta (Aß), which inhibits the synthesis of the endogenous ligand, retinoic acid (RA). This results in the counteraction of a variety of RARα-activated pathways that are key in the aetiopathology of Alzheimer's disease (AD) but which can be reversed by an RARα agonist. RARα signalling improves cognition in the Tg2576 mice, it has an anti-inflammatory effect and promotes Aß clearance by increasing insulin degrading enzyme and neprilysin activity in both microglia and neurons. In addition, RARα signalling prevents tau phosphorylation. Therefore, stimulation of the RARα signalling pathway using a synthetic agonist, by both clearing Aß and counteracting some of its toxic effects, offers therapeutic potential for the treatment of AD.
Assuntos
Doença de Alzheimer/metabolismo , Peptídeos beta-Amiloides/metabolismo , Receptor X Retinoide alfa/agonistas , Tretinoína/metabolismo , Animais , Benzoatos/farmacologia , Cognição/efeitos dos fármacos , Regulação para Baixo , Insulisina/metabolismo , Camundongos , Microglia/metabolismo , Neprilisina/metabolismo , Neurônios/metabolismo , Receptor X Retinoide alfa/metabolismo , Transdução de Sinais , Tetra-Hidronaftalenos/farmacologiaRESUMO
Alzheimer's disease is a complex disorder encompassing multiple pathological features with associated genetic and molecular culprits. However, target-based therapeutic strategies have so far proved ineffective. The aim of this study is to develop a methodology harnessing the transcriptional changes associated with Alzheimer's disease to develop a high content quantitative disease phenotype that can be used to repurpose existing drugs. Firstly, the Alzheimer's disease gene expression landscape covering severe disease stage, early pathology progression, cognitive decline and animal models of the disease has been defined and used to select a set of 153 drugs tending to oppose disease-associated changes in the context of immortalised human cancer cell lines. The selected compounds have then been assayed in the more biologically relevant setting of iPSC-derived cortical neuron cultures. It is shown that 51 of the drugs drive expression changes consistently opposite to those seen in Alzheimer's disease. It is hoped that the iPSC profiles will serve as a useful resource for drug repositioning within the context of neurodegenerative disease and potentially aid in generating novel multi-targeted therapeutic strategies.
Assuntos
Doença de Alzheimer/tratamento farmacológico , Reposicionamento de Medicamentos , Células-Tronco Pluripotentes Induzidas , Doença de Alzheimer/genética , Humanos , Células-Tronco Pluripotentes Induzidas/metabolismo , Neurônios/metabolismoRESUMO
Neuropathic pain (NP) is associated with profound gene expression alterations within the nociceptive system. DNA mechanisms, such as epigenetic remodeling and repair pathways have been implicated in NP. Here we have used a rat model of peripheral nerve injury to study the effect of a recently developed RARß agonist, C286, currently under clinical research, in NP. A 4-week treatment initiated 2 days after the injury normalized pain sensation. Genome-wide and pathway enrichment analysis showed that multiple mechanisms persistently altered in the spinal cord were restored to preinjury levels by the agonist. Concomitant upregulation of DNA repair proteins, ATM and BRCA1, the latter being required for C286-mediated pain modulation, suggests that early DNA repair may be important to prevent phenotypic epigenetic imprints in NP. Thus, C286 is a promising drug candidate for neuropathic pain and DNA repair mechanisms may be useful therapeutic targets to explore.
RESUMO
Bridgehead substituted derivatives of bicyclo[4.2.1]nonanes were synthesized and shown to be potent inhibitors of gamma-secretase. Two related series were synthesized to explore the SARs. More potent compounds were found in the non-benzofused series compared with the benzofused series. One compound from each series showed good exposure in the hepatic portal vein (HPV) following oral dosing to rats.
Assuntos
Secretases da Proteína Precursora do Amiloide/antagonistas & inibidores , Compostos Bicíclicos com Pontes/química , Compostos Bicíclicos com Pontes/farmacologia , Inibidores de Proteases/química , Inibidores de Proteases/farmacologia , Sulfonamidas/química , Sulfonamidas/farmacologia , Administração Oral , Alcanos/síntese química , Alcanos/química , Alcanos/farmacocinética , Alcanos/farmacologia , Animais , Compostos Bicíclicos com Pontes/síntese química , Compostos Bicíclicos com Pontes/farmacocinética , Linhagem Celular Tumoral , Humanos , Veia Porta/metabolismo , Inibidores de Proteases/síntese química , Inibidores de Proteases/farmacocinética , Ratos , Relação Estrutura-Atividade , Sulfonamidas/síntese química , Sulfonamidas/farmacocinéticaRESUMO
Novel benzodiazepine-containing gamma-secretase inhibitors for potential use in Alzheimer's disease have been designed that incorporate a substituted hydrocinnamide C-3 side chain. A syn combination of alpha-alkyl or aryl and beta-hydroxy or hydroxymethyl substituents was shown to give highly potent compounds. In particular, (2S,3R)-3-(3,4-difluorophenyl)-2-(4-fluorophenyl)-4-hydroxy-N-((3S)-2-oxo-5-phenyl-2,3-dihydro-1H-benzo[e][1,4]diazepin-3-yl)butyramide (34) demonstrated excellent in vitro potency (IC(50) = 0.06 nM). 34 could also be selectively methylated to give [(3)H]-28, which is of use in radioligand binding assays.
Assuntos
Benzodiazepinas/síntese química , Benzodiazepinonas/síntese química , Endopeptidases/metabolismo , Inibidores de Proteases/síntese química , Secretases da Proteína Precursora do Amiloide , Peptídeos beta-Amiloides/antagonistas & inibidores , Peptídeos beta-Amiloides/biossíntese , Ácido Aspártico Endopeptidases , Benzodiazepinas/química , Benzodiazepinas/farmacologia , Benzodiazepinonas/química , Benzodiazepinonas/farmacologia , Desenho de Fármacos , Humanos , Marcação por Isótopo , Ligantes , Inibidores de Proteases/química , Inibidores de Proteases/farmacologia , Relação Estrutura-Atividade , Células Tumorais CultivadasRESUMO
The development of potent gamma-secretase inhibitors having substituted heterocycles attached to a benzobicyclo[4.2.1]nonane core is described. This work led to the identification of [6S,9R,11R]-2',3',4',5,5',6,7,8,9,10-decahydro-2-(5-(4-fluorophenyl)-1-methylpyrazol-3-yl)-5'-(2,2,2-trifluoroethyl)spiro[6,9-methanobenzocyclooctene-11,3'-[1,2,5]thiadiazole] 1',1'-dioxide (16), which has excellent in vitro potency (0.06 nM) and which reduced amyloid-beta in APP-YAC mice with an ED(50) of 1 mg/kg (po). 16 had a good pharmacokinetic profile in three preclinical species.
Assuntos
Secretases da Proteína Precursora do Amiloide/antagonistas & inibidores , Peptídeos beta-Amiloides/antagonistas & inibidores , Ciclo-Octanos/farmacologia , Inibidores de Proteases/farmacologia , Tiadiazóis/farmacologia , Administração Oral , Animais , Ciclo-Octanos/administração & dosagem , Ciclo-Octanos/síntese química , Ciclo-Octanos/farmacocinética , Concentração Inibidora 50 , Camundongos , Inibidores de Proteases/administração & dosagem , Inibidores de Proteases/síntese química , Inibidores de Proteases/farmacocinética , Tiadiazóis/administração & dosagem , Tiadiazóis/síntese química , Tiadiazóis/farmacocinéticaRESUMO
Previously, chemistry effort on the gem-cyclohexane series of gamma-secretase inhibitors has focused on the 4-position of the cyclohexane ring. Recently chemistry has been directed towards the 3-position and substitution here has also provided compounds with high gamma-secretase activity.
Assuntos
Doença de Alzheimer/patologia , Cicloexanos/química , Endopeptidases/metabolismo , Inibidores de Proteases/química , Sulfonas/química , Doença de Alzheimer/tratamento farmacológico , Doença de Alzheimer/enzimologia , Secretases da Proteína Precursora do Amiloide , Ácido Aspártico Endopeptidases , Cicloexanos/farmacologia , Humanos , Concentração Inibidora 50 , Modelos Químicos , Inibidores de Proteases/farmacologia , Estereoisomerismo , Sulfonas/farmacologiaRESUMO
Gamma-secretase is one of the critical enzymes required for the generation of amyloid-beta peptides from the beta-amyloid precursor protein. Because amyloid-beta peptides are generally accepted to play a key role in Alzheimer disease, gamma-secretase inhibition holds the promise for a disease-modifying therapy for this neurodegenerative condition. Although recent progress has enhanced the understanding of the biology and composition of the gamma-secretase enzyme complex, less information is available on the actual interaction of various inhibitor classes with the enzyme. Here we show that the two principal classes of inhibitor described in the scientific and patent literature, aspartyl protease transition state analogue and small molecule non-transition state inhibitors, display fundamental differences in the way they interact with the enzyme. Taking advantage of a gamma-secretase enzyme overexpressing cellular system and different radiolabeled gamma-secretase inhibitors, we observed that the maximal binding of non-transition state gamma-secretase inhibitors accounts only for half the number of catalytic sites of the recombinant enzyme complex. This characteristic stoichiometry can be best accommodated with a model whereby the non-transition state inhibitors bind to a unique site at the interface of a dimeric enzyme. Subsequent competition studies confirm that this site appears to be targeted by the main classes of small molecule gamma-secretase inhibitor. In contrast, the non-steroidal anti-inflammatory drug gamma-secretase modulator sulindac sulfide displayed noncompetitive antagonism for all types of inhibitor. This finding suggests that non-steroidal anti-inflammatory drug-type gamma-secretase modulators target an alternative site on the enzyme, thereby changing the conformation of the binding sites for gamma-secretase inhibitors.
Assuntos
Secretases da Proteína Precursora do Amiloide/química , Secretases da Proteína Precursora do Amiloide/metabolismo , Anti-Inflamatórios não Esteroides/farmacologia , Ligação Competitiva , Bioquímica/métodos , Domínio Catalítico , Linhagem Celular , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos/métodos , Inibidores Enzimáticos/farmacologia , Humanos , Cinética , Ligantes , Modelos Químicos , Ligação Proteica , Sulindaco/análogos & derivados , Sulindaco/farmacologiaRESUMO
The protease gamma-secretase plays a pivotal role in the synthesis of pathogenic amyloid-beta in Alzheimer's disease (AD). Here, we report a further extension to a series of cyclohexyl sulfone-based gamma-secretase inhibitors which has allowed the preparation of highly potent compounds which also demonstrate robust Abeta(40) lowering in vivo (e.g., compound 32, MED 1mg/kg p.o. in APP-YAC mice).
Assuntos
Cicloexanos/administração & dosagem , Cicloexanos/farmacologia , Endopeptidases/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Sulfonas/administração & dosagem , Sulfonas/farmacologia , Administração Oral , Secretases da Proteína Precursora do Amiloide , Peptídeos beta-Amiloides/efeitos dos fármacos , Animais , Ácido Aspártico Endopeptidases , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Cicloexanos/química , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Camundongos , Estrutura Molecular , Fragmentos de Peptídeos/efeitos dos fármacos , Relação Estrutura-Atividade , Sulfonas/química , Fatores de TempoRESUMO
Presenilins appear to form the active center of gamma-secretase but require the presence of the integral membrane proteins nicastrin, anterior pharynx defective 1, and presenilin enhancer 2 for catalytic function. We have simultaneously overexpressed all of these polypeptides, and we demonstrate functional assembly of the enzyme complex, a substantial increase in enzyme activity, and binding of all components to a transition state analogue gamma-secretase inhibitor. Co-localization of all components can be observed in the Golgi compartment, and further trafficking of the individual constituents seems to be dependent on functional assembly. Apart from its catalytic function, gamma-secretase appears to play a role in the trafficking of the beta-amyloid precursor protein, which was changed upon reconstitution of the enzyme but unaffected by pharmacological inhibition. Because the relative molecular mass and stoichiometry of the active enzyme complex remain elusive, we performed size exclusion chromatography of solubilized gamma-secretase, which yielded evidence of a tetrameric form of the complex, yet almost completely abolished enzyme activity. Gamma-secretase activity was reconstituted upon addition of an independent size exclusion chromatography fraction of lower molecular mass and nonproteinaceous nature, which could be replaced by a brain lipid extract. The same treatment was able to restore enzyme activity after immunoaffinity purification of the gamma-secretase complex, demonstrating that lipids play a key role in preserving the catalytic activity of this protease. Furthermore, these data show that it is important to discriminate between intact, inactive gamma-secretase complexes and the active form of the enzyme, indicating the care that must be taken in the study of gamma-secretase.
Assuntos
Endopeptidases/biossíntese , Metabolismo dos Lipídeos , Glicoproteínas de Membrana/metabolismo , Peptídeo Hidrolases/metabolismo , Secretases da Proteína Precursora do Amiloide , Precursor de Proteína beta-Amiloide/metabolismo , Ácido Aspártico Endopeptidases , Sítios de Ligação , Western Blotting , Encéfalo/metabolismo , Catálise , Linhagem Celular , Membrana Celular/metabolismo , Sistema Livre de Células/metabolismo , Cromatografia , Meios de Cultura/metabolismo , Meios de Cultivo Condicionados/farmacologia , DNA Complementar/metabolismo , Dimerização , Endopeptidases/metabolismo , Complexo de Golgi/metabolismo , Humanos , Imuno-Histoquímica , Peptídeos/química , Estrutura Terciária de Proteína , Fatores de TempoRESUMO
A novel series of N-alkyl-substituted cyclic sulfamides were developed from a screening hit. Chemistries were developed which allowed surveys of N-alkyl groups and amines resulting in the identification of N-trifluoroethyl-substituted cyclic sulfamides with good in vitro and in vivo gamma-secretase activity. One compound with subnanomolar activity elicited a reduction in brain Abeta40 after oral dosing in APP-YAC mice.
Assuntos
Peptídeos beta-Amiloides/antagonistas & inibidores , Inibidores de Proteases/síntese química , Ácidos Sulfônicos/síntese química , Administração Oral , Secretases da Proteína Precursora do Amiloide , Animais , Ácido Aspártico Endopeptidases , Química Encefálica , Endopeptidases , Compostos Heterocíclicos/síntese química , Compostos Heterocíclicos/farmacologia , Concentração Inibidora 50 , Camundongos , Inibidores de Proteases/farmacologia , Relação Estrutura-Atividade , Ácidos Sulfônicos/farmacologiaRESUMO
A new series of gamma-secretase inhibitors was developed from an in-house screening hit based on a benzobicyclo[4.2.1]nonane core. Lead optimisation studies led to the development of a series of potent inhibitors and in vivo efficacy was demonstrated.