RESUMO
There are currently no approved disease-modifying osteoarthritis (OA) drugs (DMOADs). The aggrecanase ADAMTS-5 is key in the degradation of human aggrecan (AGC), a component of cartilage. Therefore, ADAMTS-5 is a promising target for the identification of DMOADs. We describe the discovery of GLPG1972/S201086, a potent and selective ADAMTS-5 inhibitor obtained by optimization of a promising hydantoin series following an HTS. Biochemical activity against rat and human ADAMTS-5 was assessed via a fluorescence-based assay. ADAMTS-5 inhibitory activity was confirmed with human aggrecan using an AGC ELISA. The most promising compounds were selected based on reduction of glycosaminoglycan release after interleukin-1 stimulation in mouse cartilage explants and led to the discovery of GLPG1972/S201086. The anticatabolic activity was confirmed in mouse cartilage explants (IC50 < 1.5 µM). The cocrystal structure of GLPG1972/S201086 with human recombinant ADAMTS-5 is discussed. GLPG1972/S201086 has been investigated in a phase 2 clinical study in patients with knee OA (NCT03595618).
Assuntos
Proteína ADAMTS5/antagonistas & inibidores , Osteoartrite/tratamento farmacológico , Proteína ADAMTS5/metabolismo , Animais , Cartilagem Articular/efeitos dos fármacos , Cartilagem Articular/metabolismo , Cães , Glicosaminoglicanos/metabolismo , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Modelos Moleculares , Osteoartrite/metabolismo , RatosRESUMO
GPR84 is a medium chain free fatty acid-binding G-protein-coupled receptor associated with inflammatory and fibrotic diseases. As the only reported antagonist of GPR84 (PBI-4050) that displays relatively low potency and selectivity, a clear need exists for an improved modulator. Structural optimization of GPR84 antagonist hit 1, identified through high-throughput screening, led to the identification of potent and selective GPR84 inhibitor GLPG1205 (36). Compared with the initial hit, 36 showed improved potency in a guanosine 5'-O-[γ-thio]triphosphate assay, exhibited metabolic stability, and lacked activity against phosphodiesterase-4. This novel pharmacological tool allowed investigation of the therapeutic potential of GPR84 inhibition. At once-daily doses of 3 and 10 mg/kg, GLPG1205 reduced disease activity index score and neutrophil infiltration in a mouse dextran sodium sulfate-induced chronic inflammatory bowel disease model, with efficacy similar to positive-control compound sulfasalazine. The drug discovery steps leading to GLPG1205 identification, currently under phase II clinical investigation, are described herein.
Assuntos
Descoberta de Drogas/métodos , Receptores Acoplados a Proteínas G/antagonistas & inibidores , Receptores Acoplados a Proteínas G/metabolismo , Acetatos/química , Acetatos/farmacologia , Regulação Alostérica/efeitos dos fármacos , Regulação Alostérica/fisiologia , Animais , Células CACO-2 , Células Cultivadas , Cães , Avaliação Pré-Clínica de Medicamentos/métodos , Feminino , Células HEK293 , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Microssomos Hepáticos/efeitos dos fármacos , Microssomos Hepáticos/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
With technologies rapidly evolving, many research institutions are now opting to invest in costly, high-quality, specialized microscopes which are shared by many researchers. As a consequence, the user may not have the ability to adapt a microscope to their specific needs and limitations in experimental design are introduced. A flexible work-horse microscopy system is a valuable tool in any laboratory to meet the diverse needs of a research team and promote innovation in experimental design. We have developed the Flexiscope; a multi-functional, adaptable, efficient and high-performance microscopy/electrophysiology system for everyday applications in a neurobiology laboratory. The core optical components are relatively constant in the three configurations described here: an upright configuration, an inverted configuration and an upright/electrophysiology configuration. We have provided a comprehensive description of the Flexiscope. We show that this method is capable of oblique infrared illumination imaging, multi-channel fluorescent imaging and automated three-dimensional scanning of larger specimens. Image quality is conserved across the three configurations of the microscope, and conversion between configurations is possible quickly and easily, while the motion control system can be repurposed to allow sub-micrometre computer-controlled micromanipulation. The Flexiscope provides similar performance and usability to commercially available systems. However, as it can be easily reconfigured for multiple roles, it can remove the need to purchase multiple microscopes, giving significant cost savings. The modular reconfigurable nature allows the user to customize the system to their specific needs and adapt/upgrade the system as challenges arise, without requiring specialized technical skills.
RESUMO
The precise cause of the bands of Fontana, striations on peripheral nerves visible to the naked eye, has been the subject of debate for hundreds of years. Some researchers have described them as reflecting the sinuous course of nerve fibres passing through nerves, and others have proposed that endoneurial collagen and sheaths surrounding nerves play a role in their appearance. We hypothesised that the bands are caused exclusively by reflection of light from the surfaces of nerve fibres travelling in phase in sinusoidal waveforms through peripheral nerves. We aligned images of obliquely illuminated nerves with confocal images of axons in those nerves, and the numbers and positions of the bands precisely matched the axonal waves. We also developed three-dimensional models of nerves with representations of the sinusoidal path of axons at their surface. We observed patterns resembling the bands of Fontana when these models were obliquely illuminated. This provides evidence that the bands of Fontana can be caused by light reflected sinusoidal path of axons alone. We subsequently describe a mechanism of band production based on our observations of both nerves and models. We report that smaller diameter nerves such as phrenic nerves and distal branches of sciatic nerves have shorter band intervals than larger nerves, such as proximal trunks of sciatic nerves, and that shorter band intervals correlate with longer axons per unit length of nerve, which suggests a greater tolerance to stretch. Inspection of banding patterns on peripheral nerves may permit prediction of axon length within nerves, and assist in the interpretation of nerve conduction data, especially in diseases where axon path has become altered.
Assuntos
Axônios/fisiologia , Nervo Frênico/citologia , Nervo Isquiático/citologia , Animais , Camundongos Endogâmicos C57BL , Ratos WistarRESUMO
Structural modification performed on a 4-methyl-4-(4-hydroxyphenyl)hydantoin series is described which resulted in the development of a new series of 4-(hydroxymethyl)diarylhydantoin analogues as potent, partial agonists of the human androgen receptor. This led to the identification of (S)-(-)-4-(4-(hydroxymethyl)-3-methyl-2,5-dioxo-4-phenylimidazolidin-1-yl)-2-(trifluoromethyl)benzonitrile ((S)-(-)-18a, GLPG0492) evaluated in vivo in a classical model of orchidectomized rat. In this model, (-)-18a exhibited anabolic activity on muscle, strongly dissociated from the androgenic activity on prostate after oral dosing. (-)-18a has very good pharmacokinetic properties, including bioavailability in rat (F > 50%), and is currently under evaluation in phase I clinical trials.
Assuntos
Androgênios/síntese química , Hidantoínas/síntese química , Anabolizantes/síntese química , Anabolizantes/química , Anabolizantes/farmacologia , Antagonistas de Receptores de Andrógenos/síntese química , Antagonistas de Receptores de Andrógenos/química , Antagonistas de Receptores de Andrógenos/farmacologia , Androgênios/química , Androgênios/farmacologia , Animais , Disponibilidade Biológica , Agonismo Parcial de Drogas , Células HeLa , Humanos , Hidantoínas/química , Hidantoínas/farmacologia , Masculino , Modelos Moleculares , Conformação Molecular , Músculo Esquelético/anatomia & histologia , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/metabolismo , Orquiectomia , Tamanho do Órgão/efeitos dos fármacos , Próstata/anatomia & histologia , Próstata/efeitos dos fármacos , Próstata/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores Androgênicos/genética , Receptores Androgênicos/metabolismo , Estereoisomerismo , Relação Estrutura-Atividade , Ativação Transcricional/efeitos dos fármacosRESUMO
We previously reported the synthesis and the antimycobacterial activity of 4-(7,7-dimethyl-7H-furo[3,2-f]chromen-2-yl)pyridine. From this result, we sought to design simple synthetic accesses to related structures allowing the preparation of a diverse set of analogues. Two approaches were investigated. From 3-(2-bromo-7,7-dimethyl-8,9-dihydro-7H-furo[3,2-f]chromen-1-yl)propyl acetate, we prepared 2-arylated derivatives via Suzuki-Miyaura reactions between this bromine-bearing compound and various arylboronates. Moreover, and even more simple, we prepared the ((6-hydroxy-2,2,7,8-tetramethylchroman-5-yl)methyl)triphenylphosphonium salt via a selective bromination of 2,2,5,7,8-pentamethylchroman-6-ol. From this salt, a two stage Wittig reaction with an array of activated acids allowed the quick preparation of many analogues. The biological evaluation of the effect of these compounds on the growth of Mycobacterium bovis as well as Mycobacterium tuberculosis pointed out a fourfold improvement of the antimycobacterial properties for one of the compounds made. However, the many analogues which inhibited the growth of M. tuberculosis in the 0.6-5 microg/mL range turned out to be also cytotoxic on VERO cells growth at the same concentration range.
Assuntos
Antibacterianos/síntese química , Antibacterianos/farmacologia , Benzopiranos/síntese química , Benzopiranos/farmacologia , Furanos/síntese química , Furanos/farmacologia , Mycobacterium bovis/efeitos dos fármacos , Mycobacterium tuberculosis/efeitos dos fármacos , Animais , Antibacterianos/química , Antifúngicos/síntese química , Antifúngicos/química , Antifúngicos/farmacologia , Benzopiranos/química , Candida tropicalis/efeitos dos fármacos , Chlorocebus aethiops , Relação Dose-Resposta a Droga , Desenho de Fármacos , Furanos/química , Testes de Sensibilidade Microbiana , Estrutura Molecular , Mycobacterium bovis/crescimento & desenvolvimento , Mycobacterium tuberculosis/crescimento & desenvolvimento , Estereoisomerismo , Células VeroRESUMO
From the structure of 3,3-dimethyl-3H-benzofuro[3,2-f][1]-benzopyran, a selective in vitro inhibitor of mycobacterial growth, we have undertaken a structure-activity relationship investigation. We wish to report here our results on the use of [2+3] cycloadditions between 2-formylbenzoquinone and various enol derivatives to give various 4-formyl-5-hydroxy benzofurans. In the next step, an ytterbium triflate-catalysed reaction with 2-methylpropene allowed the preparation of various original furo[3,2-f]chromenes derivatives. Their biological evaluation on the growth of Mycobacterium smegmatis as well as Mycobacterium tuberculosis pointed out that some analogues were four times more active than the initial hit.
Assuntos
Antibacterianos/síntese química , Antibacterianos/farmacologia , Benzofuranos/síntese química , Benzofuranos/farmacologia , Benzopiranos/síntese química , Benzopiranos/farmacologia , Mycobacterium smegmatis/efeitos dos fármacos , Mycobacterium tuberculosis/efeitos dos fármacos , Antibacterianos/química , Benzofuranos/química , Benzopiranos/química , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos , Espectroscopia de Ressonância Magnética/métodos , Testes de Sensibilidade Microbiana , Estrutura Molecular , Mycobacterium smegmatis/crescimento & desenvolvimento , Mycobacterium tuberculosis/crescimento & desenvolvimento , Estereoisomerismo , Relação Estrutura-AtividadeRESUMO
A series of new 7-monosubstituted and 3,7-disubstituted indazoles have been prepared and evaluated as inhibitors of nitric oxide synthases (NOS). 1H-indazole-7-carbonitrile (6) was found equipotent to 7-nitro-1H-indazole (1) and demonstrated preference for constitutive NOS over inducible NOS. By contrast, 1H-indazole-7-carboxamide (8) was slightly less potent but demonstrated a surprising selectivity for the neuronal NOS. Further substitution of 6 by a Br-atom at carbon-3 of the heterocycle enhanced 10-fold the inhibitory effects. Inhibition of NO formation by 6 appeared to be competitive versus both substrate and the cofactor (6R)-5,6,7,8-tetrahydro-l-biopterin (H(4)B). In close analogies with 1, compound 6 strongly inhibited the NADPH oxidase activity of nNOS and induced a spin state transition of the heme-Fe(III). Our results are explained with the help of the X-ray structures that identified key-features for binding of 1 at the active site of NOS.
Assuntos
Indazóis/síntese química , Indazóis/farmacologia , Óxido Nítrico Sintase/antagonistas & inibidores , Animais , Sítios de Ligação , Catálise , Linhagem Celular , Indazóis/metabolismo , Camundongos , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase/metabolismo , Óxido Nítrico Sintase Tipo I/química , Óxido Nítrico Sintase Tipo I/metabolismo , Óxido Nítrico Sintase Tipo II/química , Óxido Nítrico Sintase Tipo III/química , Espectrofotometria Ultravioleta , Relação Estrutura-AtividadeRESUMO
Reactions of PH(3) and commercially available H(2)C=CHR(f) (R(f6)(/)(8)(/)(10) = (CF(2))(5)CF(3)/(CF(2))(7)CF(3)/(CF(2))(9)CF(3)) give, in two-stage processes conducted with free radical initiators (AIBN, VAZO; 80-90 degrees C), the phosphines P(CH(2)CH(2)R(f))(3) (1-3; 63-75%). Analogous reactions with H(2)C=CHCH(2)R(f8) (7) and H(2)C=CHCH(2)CH(2)R(f8) (10) give P(CH(2)CH(2)CH(2)R(f8))(3) (4, 73%) and P(CH(2)CH(2)CH(2)CH(2)R(f8))(3) (5, 66%), in which the phosphorus is increasingly insulated from the electronegative R(f) moiety. The alkenes 7 and 10 are prepared from Bu(3)SnCH(2)CH=CH(2) and IR(f8) (hnu, CH(2)Cl(2), 81%) or ICH(2)R(f8) (VAZO, refluxing CF(3)C(6)H(5), 56%). The reaction of 1 and H(2)O(2) gives O=P(CH(2)CH(2)R(f6))(3) (6, 88%), which can be reduced with HSiCl(3) to 1. Partition coefficients (CF(3)C(6)F(11)/toluene, 27 degrees C) range from 98.8:1.2 (1, 4) through 98.9:1.1 (5) to >99.7:<0.3 (2, 3, 6). Crystals of 4 diffract poorly, but a packing motif that maximizes interactions between R(f) segments is evident.