RESUMEN
Compound 21 (AM432) was identified as a potent and selective antagonist of the DP(2) receptor (CRTH2). Modification of a bi-aryl core identified a series of tri-aryl antagonists of which compound 21 proved a viable clinical candidate. AM432 shows excellent potency in a human whole blood eosinophil shape change assay with prolonged incubation, a comparatively long off-rate from the DP(2) receptor, excellent pharmacokinetics in dog and in vivo activity in two mouse models of inflammatory disease after oral dosing.
Asunto(s)
Fenilacetatos/química , Piridinas/química , Receptores Inmunológicos/antagonistas & inhibidores , Receptores de Prostaglandina/antagonistas & inhibidores , Administración Oral , Animales , Modelos Animales de Enfermedad , Perros , Eosinófilos/efectos de los fármacos , Eosinófilos/inmunología , Humanos , Inflamación/tratamiento farmacológico , Ratones , Fenilacetatos/farmacocinética , Fenilacetatos/uso terapéutico , Piridinas/farmacocinética , Piridinas/uso terapéutico , Receptores Inmunológicos/metabolismo , Receptores de Prostaglandina/metabolismoRESUMEN
The incorporation of synthetic nucleoside analogues into DNA duplexes provides a unique opportunity to probe both structure and function of nucleic acids. We used 1H and 19F NMR and molecular dynamics calculations to determine the solution structures of two similar DNA decamer duplexes, one containing a central G-T mismatched or "wobble" base pair, and one in which the thymine in this base pair is replaced by difluorotoluene (a thymine isostere) creating a G-F pair. Here, we show that the non-hydrogen-bonding G-F pair stacks relatively well into the helix and that the distortions caused by each non-Watson-Crick G-T or G-F base pair are quite localized to a three base pair site around the mismatch. A detailed structural analysis reveals that the absence of hydrogen bonding introduces more dynamic motion into the G-F pair relative to G-T and permits the G-F pair to exhibit stacking and conformational features characteristic of both a Watson-Crick base pair (on the guanine containing strand) and a wobble base pair (on the strand containing the difluorotoluene). We used these results to posit a rationale for recognition and repair of mismatch sites in DNA.
Asunto(s)
ADN/química , Guanina/química , Tolueno/análogos & derivados , Emparejamiento Base , Reparación del ADN , Enlace de Hidrógeno , Espectroscopía de Resonancia Magnética , Oligonucleótidos/síntesis química , Oligonucleótidos/química , Teoría Cuántica , Soluciones , Tolueno/químicaRESUMEN
Porphyrinmaleimides were synthesized and characterized. The thiol-containing amino acid L-cysteine reacted with 58% yield with these porphyrins to form bioconjugate adducts. The new thiol-active reagents were labeled cytoplasmic cysteine 140 and 316 in rhodopsin (Rh), a G protein coupled receptor (GPCR).