RESUMEN
Activation of the cGAS-STING pathway plays a key role in the innate immune response to cancer through Type-1 Interferon (IFN) production and T cell priming. Accumulation of cytosolic double-stranded DNA (dsDNA) within tumor cells and dying cells is recognized by the DNA sensor cyclic GMP-AMP synthase (cGAS) to create the secondary messenger cGAMP, which in turn activates STING (STimulator of INterferon Genes), resulting in the subsequent expression of IFN-related genes. This process is regulated by Three-prime Repair EXonuclease 1 (TREX1), a 3' â 5' exonuclease that degrades cytosolic dsDNA, thereby dampening activation of the cGAS-STING pathway, which in turn diminishes immunostimulatory IFN secretion. Here, we characterize the activity of VB-85680, a potent small-molecule inhibitor of TREX1. We first demonstrate that VB-85680 inhibits TREX1 exonuclease activity in vitro in lysates from both human and mouse cell lines. We then show that treatment of intact cells with VB-85680 results in activation of downstream STING signaling, and activation of IFN-stimulated genes (ISGs). THP1-Dual™ cells cultured under low-serum conditions exhibited an enhanced ISG response when treated with VB-85680 in combination with exogenous DNA. Collectively, these findings suggest the potential of a TREX1 exonuclease inhibitor to work in combination with agents that generate cytosolic DNA to enhance the acquisition of the anti-tumor immunity widely associated with STING pathway activation.
Asunto(s)
Exodesoxirribonucleasas , Fosfoproteínas , Exodesoxirribonucleasas/metabolismo , Humanos , Fosfoproteínas/metabolismo , Ratones , Animales , Nucleotidiltransferasas/metabolismo , Nucleotidiltransferasas/antagonistas & inhibidores , Proteínas de la Membrana/metabolismo , Regulación hacia Arriba/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Interferones/metabolismo , Inmunidad Innata/efectos de los fármacosRESUMEN
Synthesis and structure-activity relationships (SAR) of a novel series of benzodiazepinedione-based inhibitors of Clostridium difficile toxin B (TcdB) are described. Compounds demonstrating low nanomolar affinity for TcdB, and which possess improved stability in mouse plasma vs. earlier compounds from this series, have been identified. Optimized compound 11d demonstrates a good pharmacokinetic (PK) profile in mouse and hamster and is efficacious in a hamster survival model of Clostridium difficile infection.
Asunto(s)
Antibacterianos/síntesis química , Proteínas Bacterianas/antagonistas & inhibidores , Toxinas Bacterianas/antagonistas & inhibidores , Benzodiazepinas/química , Administración Oral , Animales , Antibacterianos/farmacocinética , Antibacterianos/uso terapéutico , Proteínas Bacterianas/metabolismo , Toxinas Bacterianas/metabolismo , Benzodiazepinas/farmacocinética , Benzodiazepinas/uso terapéutico , Células CHO , Clostridioides difficile/metabolismo , Infecciones por Clostridium/tratamiento farmacológico , Infecciones por Clostridium/veterinaria , Cricetinae , Cricetulus , Semivida , Ratones , Relación Estructura-ActividadRESUMEN
Clostridium difficile infection (CDI) is the leading cause of hospital-acquired infectious diarrhea, with significant morbidity, mortality, and associated health care costs. The major risk factor for CDI is antimicrobial therapy, which disrupts the normal gut microbiota and allows C. difficile to flourish. Treatment of CDI with antimicrobials is generally effective in the short term, but recurrent infections are frequent and problematic, indicating that improved treatment options are necessary. Symptoms of disease are largely due to two homologous toxins, TcdA and TcdB, which are glucosyltransferases that inhibit host Rho GTPases. As the normal gut microbiota is an important component of resistance to CDI, our goal was to develop an effective nonantimicrobial therapy. Here, we report a highly potent small-molecule inhibitor (VB-82252) of TcdA and TcdB. This compound inhibits the UDP-glucose hydrolysis activity of TcdB and protects cells from intoxication after challenge with either toxin. Oral dosing of the inhibitor prevented inflammation in a murine intrarectal toxin challenge model. In a murine model of recurrent CDI, the inhibitor reduced weight loss and gut inflammation during acute disease and did not cause the recurrent disease that was observed with vancomycin treatment. Lastly, the inhibitor demonstrated efficacy similar to that of vancomycin in a hamster disease model. Overall, these results demonstrate that small-molecule inhibition of C. difficile toxin UDP-glucose hydrolysis activity is a promising nonantimicrobial approach to the treatment of CDI.
Asunto(s)
Antibacterianos/uso terapéutico , Infecciones por Clostridium/tratamiento farmacológico , Uridina Difosfato Glucosa/metabolismo , Animales , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Toxinas Bacterianas/genética , Toxinas Bacterianas/metabolismo , Línea Celular , Supervivencia Celular , Clostridioides difficile/efectos de los fármacos , Clostridioides difficile/patogenicidad , Infecciones por Clostridium/metabolismo , Colon/microbiología , Cricetinae , Humanos , Hidrólisis , RatonesRESUMEN
The discovery, synthesis and preliminary structure-activity relationship (SAR) of a novel class of inhibitors of Clostridium difficile (C. difficile) toxin B (TcdB) is described. A high throughput screening (HTS) campaign resulted in the identification of moderately active screening hits 1-5 the most potent of which was compound 1 (IC50â¯=â¯0.77⯵M). In silico docking of an early analog offered suggestions for structural modification which resulted in the design and synthesis of highly potent analogs 13j(IC50â¯=â¯1â¯nM) and 13â¯l(IC50â¯=â¯7â¯nM) which were chosen as leads for further optimization.
Asunto(s)
Antibacterianos/farmacología , Proteínas Bacterianas/antagonistas & inhibidores , Toxinas Bacterianas/antagonistas & inhibidores , Clostridioides difficile/efectos de los fármacos , Nucleotidasas/antagonistas & inhibidores , Animales , Antibacterianos/síntesis química , Antibacterianos/química , Antibacterianos/farmacocinética , Apoptosis/efectos de los fármacos , Células CHO , Cricetulus , Estabilidad de Medicamentos , Enterotoxinas/antagonistas & inhibidores , Ensayos Analíticos de Alto Rendimiento , Humanos , Ratones , Pruebas de Sensibilidad Microbiana , Microsomas Hepáticos/metabolismo , Simulación del Acoplamiento Molecular , Estructura Molecular , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
Synthesis and structure-activity relationships (SAR) of a novel series of vasopressin V(1b) antagonists are described. 2-(6-Aminomethylaryl-2-aryl-4-oxo-quinazolin-3(4H)-yl)acetamide have been identified with low nanomolar affinity for the V(1b) receptor and good selectivity with respect to related receptors V(1a), V(2) and OT. Optimised compound 16 shows a good pharmacokinetic profile and activity in a mechanistic model of HPA dysfunction.
Asunto(s)
Acetamidas/síntesis química , Antagonistas de los Receptores de Hormonas Antidiuréticas , Sistema Hipotálamo-Hipofisario/efectos de los fármacos , Quinazolinonas/síntesis química , Quinazolinonas/farmacología , Acetamidas/química , Acetamidas/farmacología , Animales , Células CACO-2 , Humanos , Concentración 50 Inhibidora , Masculino , Estructura Molecular , Quinazolinonas/química , Ratas , Ratas Wistar , Relación Estructura-ActividadRESUMEN
Synthesis and structure-activity relationships (SAR) of a novel series of vasopressin V(1b) (V(3)) antagonists are described. 2-(4-Oxo-2-aryl-quinazolin-3(4H)-yl)acetamides have been identified with low nanomolar affinity for the V(1b) receptor and good selectivity with respect to related receptors V(1a), V(2) and oxytocin (OT). Optimised compound 12j demonstrates a good pharmacokinetic profile and activity in a mechanistic model of HPA dysfunction.
Asunto(s)
Antagonistas de los Receptores de Hormonas Antidiuréticas , Quinazolinas/síntesis química , Quinazolinas/farmacología , Animales , Humanos , Quinazolinas/química , Quinazolinas/farmacocinética , Ratas , Relación Estructura-ActividadRESUMEN
The discovery, synthesis, and preliminary structure-activity relationship (SAR) of a novel class of vasopressin V3 (V1b) receptor antagonists is described. Compound 1, identified by high throughput screening of a diverse, three million-member compound collection, prepared using ECLiPS technology, had good activity in a V3 binding assay (IC50=0.20 microM), but less than desirable physicochemical properties. Optimization of compound 1 yielded potent analogs 19 (IC50=0.31 microM) and 24 (IC50=0.12 microM) with improved drug-like characteristics.
Asunto(s)
Acetamidas/química , Acetamidas/farmacología , Antagonistas de los Receptores de Hormonas Antidiuréticas , Quinazolinas/química , Quinazolinas/farmacología , Receptores de Vasopresinas/metabolismo , Acetamidas/síntesis química , Animales , Trastorno Depresivo/tratamiento farmacológico , Humanos , Quinazolinas/síntesis química , Ratas , Relación Estructura-ActividadRESUMEN
The discovery, synthesis and preliminary structure-activity relationships (SARs) of a novel class of CB1 antagonists is described. Initial optimization of benzimidazole-based screening hit 4 led to the identification of 'inverted' indole-based lead compound 18c with improved properties versus compound 4 including reduced AlogP, improved microsomal stability and improved aqueous solubility. Compound 18c demonstrates in vivo CB1 antagonist efficacy (CB1 agonist induced hypothermia model) and is orally bioavailable in rat.
Asunto(s)
Bencimidazoles/química , Bencimidazoles/farmacología , Indoles/química , Indoles/farmacología , Receptor Cannabinoide CB1/antagonistas & inhibidores , Receptor Cannabinoide CB1/metabolismo , Animales , Bencimidazoles/metabolismo , Bencimidazoles/farmacocinética , Humanos , Hipotermia/inducido químicamente , Hipotermia/tratamiento farmacológico , Indoles/metabolismo , Indoles/farmacocinética , Masculino , Ratones , Microsomas Hepáticos/metabolismo , Obesidad/tratamiento farmacológico , Ratas , Ratas Wistar , Solubilidad , Relación Estructura-ActividadRESUMEN
A novel class of Janus tyrosine kinase 3 (JAK3) inhibitors based on a 2-benzimidazoylpurinone core structure is described. Through substitution of the benzimidazoyl moiety and optimization of the N-9 substituent of the purinone, compound 24 was identified incorporating a chroman-based functional group. Compound 24 shows excellent kinase activity, good oral bioavailability and demonstrates efficacy in an acute mechanistic mouse model through inhibition of interleukin-2 (IL-2) induced interferon-gamma (INF-gamma) production.
Asunto(s)
Bencimidazoles/farmacología , Inhibidores Enzimáticos/farmacología , Janus Quinasa 3/antagonistas & inhibidores , Purinas/farmacología , Animales , Bencimidazoles/síntesis química , Bencimidazoles/química , Relación Dosis-Respuesta a Droga , Diseño de Fármacos , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/química , Interferón gamma/biosíntesis , Interleucina-2/antagonistas & inhibidores , Ratones , Modelos Animales , Modelos Moleculares , Estructura Molecular , Purinas/síntesis química , Purinas/química , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
The discovery, synthesis and preliminary SAR of a novel class of non-peptidic antagonists of the alpha(v)-integrins alpha(v)beta(3) and alpha(v)beta(5) is described. High-throughput screening of an extensive series of ECLiPStrade mark compound libraries led to the identification of compound 1 as a dual inhibitor of the alpha(v)-integrins alpha(v)beta(3) and alpha(v)beta(5). Optimization of compound 1 involving, in part, introduction of two novel constraints led to the discovery of compounds 15a and 15b with reduced PSA and much improved potency for both the alpha(v)beta(3) and alpha(v)beta(5) integrins. Compounds 15a and 15b were shown to have promising activity in functional cellular assays and compound 15a also exhibited a promising Caco-2 permeability profile.