RESUMO
Starting from the micromolar 8-quinoline carboxamide high-throughput screening hit 1a, a systematic exploration of the structure-activity relationships (SAR) of the 4-, 6-, and 8-substituents of the quinoline ring resulted in the identification of approximately 10-100-fold more potent human CD38 inhibitors. Several of these molecules also exhibited pharmacokinetic parameters suitable for in vivo animal studies, including low clearances and decent oral bioavailability. Two of these CD38 inhibitors, 1ah and 1ai, were shown to elevate NAD tissue levels in liver and muscle in a diet-induced obese (DIO) C57BL/6 mouse model. These inhibitor tool compounds will enable further biological studies of the CD38 enzyme as well as the investigation of the therapeutic implications of NAD enhancement in disease models of abnormally low NAD.
Assuntos
ADP-Ribosil Ciclase 1/antagonistas & inibidores , Amidas/química , Aminoquinolinas/química , NAD/metabolismo , Quinolinas/química , Amidas/síntese química , Amidas/farmacologia , Aminoquinolinas/síntese química , Aminoquinolinas/farmacologia , Animais , Disponibilidade Biológica , Cristalografia por Raios X , Humanos , Hidrólise , Fígado/metabolismo , Membranas Artificiais , Camundongos Endogâmicos C57BL , Modelos Moleculares , Músculo Esquelético/metabolismo , Obesidade/metabolismo , Permeabilidade , Conformação Proteica , Quinolinas/síntese química , Quinolinas/farmacologia , Estereoisomerismo , Relação Estrutura-AtividadeRESUMO
Nicotinamide adenine dinucleotide (NAD+) is a key cofactor required for essential metabolic oxidation-reduction reactions. It also regulates various cellular activities, including gene expression, signaling, DNA repair and calcium homeostasis. Intracellular NAD+ levels are tightly regulated and often respond rapidly to nutritional and environmental changes. Numerous studies indicate that elevating NAD+ may be therapeutically beneficial in the context of numerous diseases. However, the role of NAD+ on skeletal muscle exercise performance is poorly understood. CD38, a multi-functional membrane receptor and enzyme, consumes NAD+ to generate products such as cyclic-ADP-ribose. CD38 knockout mice show elevated tissue and blood NAD+ level. Chronic feeding of high-fat, high-sucrose diet to wild type mice leads to exercise intolerance and reduced metabolic flexibility. Loss of CD38 by genetic mutation protects mice from diet-induced metabolic deficit. These animal model results suggest that elevation of tissue NAD+ through genetic ablation of CD38 can profoundly alter energy homeostasis in animals that are maintained on a calorically-excessive Western diet.
Assuntos
ADP-Ribosil Ciclase 1/genética , ADP-Ribosil Ciclase 1/metabolismo , Dieta Ocidental/efeitos adversos , Doenças Metabólicas/genética , Doenças Metabólicas/metabolismo , Condicionamento Físico Animal/fisiologia , ADP-Ribosil Ciclase/metabolismo , Animais , ADP-Ribose Cíclica/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Músculo Esquelético/metabolismo , NAD/metabolismo , OxirreduçãoRESUMO
A series of thiazoloquin(az)olinones were synthesized and found to have potent inhibitory activity against CD38. Several of these compounds were also shown to have good pharmacokinetic properties and demonstrated the ability to elevate NAD levels in plasma, liver, and muscle tissue. In particular, compound 78c was given to diet induced obese (DIO) C57Bl6 mice, elevating NAD > 5-fold in liver and >1.2-fold in muscle versus control animals at a 2 h time point. The compounds described herein possess the most potent CD38 inhibitory activity of any small molecules described in the literature to date. The inhibitors should allow for a more detailed assessment of how NAD elevation via CD38 inhibition affects physiology in NAD deficient states.
Assuntos
ADP-Ribosil Ciclase 1/antagonistas & inibidores , Quinolonas/química , Quinolonas/farmacologia , Tiazóis/química , Tiazóis/farmacologia , ADP-Ribosil Ciclase 1/metabolismo , Animais , Linhagem Celular , Cães , Descoberta de Drogas , Humanos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Camundongos Endogâmicos C57BL , Simulação de Acoplamento Molecular , Músculos/efeitos dos fármacos , Músculos/metabolismo , NAD/análise , NAD/sangue , NAD/metabolismo , Obesidade/tratamento farmacológico , Obesidade/metabolismo , Quinolonas/síntese química , Quinolonas/farmacocinética , Tiazóis/síntese química , Tiazóis/farmacocinéticaRESUMO
The metalloprotease ADAMTS-5 is considered a potential target for the treatment of osteoarthritis. To identify selective inhibitors of ADAMTS-5, we employed encoded library technology (ELT), which enables affinity selection of small molecule binders from complex mixtures by DNA tagging. Selection of ADAMTS-5 against a four-billion member ELT library led to a novel inhibitor scaffold not containing a classical zinc-binding functionality. One exemplar, (R)-N-((1-(4-(but-3-en-1-ylamino)-6-(((2-(thiophen-2-yl)thiazol-4-yl)methyl)amino)-1,3,5-triazin-2-yl)pyrrolidin-2-yl)methyl)-4-propylbenzenesulfonamide (8), inhibited ADAMTS-5 with IC(50) = 30 nM, showing >50-fold selectivity against ADAMTS-4 and >1000-fold selectivity against ADAMTS-1, ADAMTS-13, MMP-13, and TACE. Extensive SAR studies showed that potency and physicochemical properties of the scaffold could be further improved. Furthermore, in a human osteoarthritis cartilage explant study, compounds 8 and 15f inhibited aggrecanase-mediated (374)ARGS neoepitope release from aggrecan and glycosaminoglycan in response to IL-1ß/OSM stimulation. This study provides the first small molecule evidence for the critical role of ADAMTS-5 in human cartilage degradation.
Assuntos
Proteínas ADAM/antagonistas & inibidores , Cartilagem Articular/efeitos dos fármacos , Bases de Dados de Compostos Químicos , Osteoartrite/patologia , Sulfonamidas/síntese química , Triazinas/síntese química , Proteína ADAMTS5 , Agrecanas/metabolismo , Animais , Cartilagem Articular/metabolismo , Cartilagem Articular/patologia , Endopeptidases/metabolismo , Epitopos , Glicosaminoglicanos/metabolismo , Humanos , Técnicas In Vitro , Masculino , Pessoa de Meia-Idade , Osteoartrite/tratamento farmacológico , Ratos , Ratos Sprague-Dawley , Bibliotecas de Moléculas Pequenas , Estereoisomerismo , Relação Estrutura-Atividade , Sulfonamidas/farmacocinética , Sulfonamidas/farmacologia , Triazinas/farmacocinética , Triazinas/farmacologiaRESUMO
We report the synthesis and in vitro activity of a series of novel substituted N-{3-[(1,1-dioxido-1,2-benzothiazol-3-yl)(phenyl)amino]propyl}benzamide analogs. These analogs showed potent inhibitory activity against Kv1.3. Several demonstrated similar potency to the known Kv1.3 inhibitor PAP-1 when tested under the IonWorks patch clamp assay conditions. Two compounds 13i and 13rr were advanced further as potential tool compounds for in vivo validation studies.