RESUMO
Therapies that promote neuroprotection and axonal survival by enhancing myelin regeneration are an unmet need to prevent disability progression in multiple sclerosis. Numerous potentially beneficial compounds have originated from phenotypic screenings but failed in clinical trials. It is apparent that current cell- and animal-based disease models are poor predictors of positive treatment options, arguing for novel experimental approaches. Here we explore the experimental power of humanized zebrafish to foster the identification of pro-remyelination compounds via specific inhibition of GPR17. Using biochemical and imaging techniques, we visualize the expression of zebrafish (zf)-gpr17 during the distinct stages of oligodendrocyte development, thereby demonstrating species-conserved expression between zebrafish and mammals. We also demonstrate species-conserved function of zf-Gpr17 using genetic loss-of-function and rescue techniques. Finally, using GPR17-humanized zebrafish, we provide proof of principle for in vivo analysis of compounds acting via targeted inhibition of human GPR17. We anticipate that GPR17-humanized zebrafish will markedly improve the search for effective pro-myelinating pharmacotherapies.
Assuntos
Oligodendroglia , Pró-Fármacos , Animais , Humanos , Peixe-Zebra/metabolismo , Pró-Fármacos/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Diferenciação Celular , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/metabolismo , Modelos Animais de Doenças , Mamíferos/metabolismoRESUMO
A multi-disciplinary approach was used to identify the first pharmacophore model for KCC2 blockers: several physico-chemical studies such as XRD and NMR were combined to molecular modelling techniques, SAR analysis and synthesis of constrained analogues in order to determine a minimal conformational space regrouping few potential bioactive conformations. These conformations were further compared to the conformational space of a different series of KCC2 blockers in order to identify the common pharmacophoric features. The synthesis of more potent analogues in this second series confirmed the usefulness of this KCC2 blocker pharmacophore model.
Assuntos
Anticonvulsivantes/síntese química , Bloqueadores dos Canais de Potássio/síntese química , Prolina/análogos & derivados , Prolina/síntese química , Simportadores/antagonistas & inibidores , Animais , Anticonvulsivantes/farmacologia , Linhagem Celular Tumoral , Furosemida/farmacologia , Ensaios de Triagem em Larga Escala , Humanos , Transporte de Íons/efeitos dos fármacos , Espectroscopia de Ressonância Magnética , Modelos Moleculares , Conformação Molecular , Bloqueadores dos Canais de Potássio/farmacologia , Prolina/farmacologia , Ratos , Relação Estrutura-Atividade , Simportadores/metabolismo , Difração de Raios X , Cotransportadores de K e Cl-RESUMO
The discovery and optimization of a novel class of selective submicromolar KCC2 blockers is described. Details of synthesis and SAR are given together with ADME properties of selected compounds. A methylsulfone residue on the R(1) phenyl group improved the overall general profile of these prolinate derivatives.
Assuntos
Prolina/análogos & derivados , Simportadores/antagonistas & inibidores , Animais , Prolina/química , Prolina/farmacologia , Ratos , Estereoisomerismo , Relação Estrutura-Atividade , Cotransportadores de K e Cl-RESUMO
The discovery and optimization of a novel class of potent CCR3 antagonists is described. Details of synthesis and SAR are given together with some ADME properties of selected compounds. An optimal balance between activities, physicochemical properties, and in vitro metabolic stability was reached by the proper choice of substituents.
Assuntos
Piperidinas/farmacologia , Receptores de Quimiocinas/antagonistas & inibidores , Humanos , Piperidinas/síntese química , Piperidinas/química , Receptores CCR3 , Relação Estrutura-AtividadeRESUMO
The synthesis and structure-activity relationships against the C3a receptor of a series of substituted aminopiperidine derivatives are reported. DMPK properties and functional activities of selected compounds are described. The compounds obtained are the first non-arginine ligands of C3aR.
Assuntos
Aminas/química , Ativação do Complemento/efeitos dos fármacos , Proteínas de Membrana/metabolismo , Piperidinas/farmacologia , Receptores de Complemento/metabolismo , Animais , Ligantes , Camundongos , Camundongos Endogâmicos BALB C , Piperidinas/síntese química , Ligação Proteica , Proteínas Serina-Treonina Quinases/metabolismo , Relação Estrutura-AtividadeRESUMO
Amido derivatives 10-18 of the corresponding oxyamines were synthesised as melatoninergic ligands by the reaction of hydroxyphtalimide with the halogeno derivatives or the corresponding alcohols using Mitsunobu reaction conditions. The affinity of the compounds for chicken brain melatonin receptors and recombinant human MT(1) and MT(2) receptors was evaluated using 2-[125I]-iodomelatonin as the radioligand. Overall, the introduction of an oxygen atom in the amido chain was not a favourable parameter as the compounds were less potent than the corresponding deoxy derivatives. However, nanomolar compounds were obtained with the arylethyloxy derivatives (13c (R'=nPr), chicken brain, hMT(1), hMT(2), K(i) values: 4.8, 3.86, 2.4 nM, respectively) and the 2,7-dimethoxynaphthalene derivatives (17c (R'=nPr), chicken brain, hMT(1), hMT(2), K(i) values: 0.04, 0.13, 0.1 nM, respectively). The functional activity of these compounds was evaluated by the aggregation of melanophores in Xenopus laevis tadpoles and the potency was related to the affinity of the molecules for melatonin receptors. The compounds were found to be full agonists and compound 17a was 20-fold more potent than melatonin in this bioassay.
Assuntos
Alcanos/síntese química , Alcanos/farmacologia , Amidas/síntese química , Amidas/farmacologia , Melatonina/análogos & derivados , Melatonina/química , Animais , Ligação Competitiva/efeitos dos fármacos , Química Encefálica/efeitos dos fármacos , Galinhas , Feminino , Humanos , Larva , Ligantes , Espectroscopia de Ressonância Magnética , Masculino , Melatonina/metabolismo , Receptores de Superfície Celular/efeitos dos fármacos , Receptores Citoplasmáticos e Nucleares/efeitos dos fármacos , Receptores de Melatonina , Proteínas Recombinantes/efeitos dos fármacos , Relação Estrutura-Atividade , Xenopus laevisRESUMO
A series of 5-substituted 2-benzoylaminobenzoic acids has been synthesized and assayed for PPARalpha/gamma activity. Both dual activators and selective PPARgamma agonists have been identified. This class of compounds was shown to activate the PPARgamma receptor through interaction with a novel binding site.