RESUMO
Androgen receptor (AR) is a ligand-activated transcription factor that plays a pivotal role in the development and progression of many severe diseases such as prostate cancer, muscle atrophy, and osteoporosis. Binding of ligands to AR triggers the conformational changes in AR that may affect the recruitment of coactivators and downstream response of AR signaling pathway. Therefore, AR ligands have great potential to treat these diseases. In this study, we searched for novel AR ligands by performing a docking-based virtual screening (VS) on the basis of the crystal structure of the AR ligand binding domain (LBD) in complex with its agonist. A total of 58 structurally diverse compounds were selected and subjected to LBD affinity assay, with five of them (HBP1-3, HBP1-17, HBP1-38, HBP1-51, and HBP1-58) exhibiting strong binding to AR-LBD. The IC values of HBP1-51 and HBP1-58 are 3.96 µM and 4.92 µM, respectively, which are even lower than that of enzalutamide (Enz, IC = 13.87 µM), a marketed second-generation AR antagonist. Further bioactivity assays suggest that HBP1-51 is an AR agonist, whereas HBP1-58 is an AR antagonist. In addition, molecular dynamics (MD) simulations and principal components analysis (PCA) were carried out to reveal the binding principle of the newly-identified AR ligands toward AR. Our modeling results indicate that the conformational changes of helix 12 induced by the bindings of antagonist and agonist are visibly different. In summary, the current study provides a highly efficient way to discover novel AR ligands, which could serve as the starting point for development of new therapeutics for AR-related diseases.
Assuntos
Humanos , Masculino , Antagonistas de Receptores de Andrógenos , Farmacologia , Androgênios , Metabolismo , Farmacologia , Bioensaio , Linhagem Celular Tumoral , Descoberta de Drogas , Métodos , Ligantes , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Feniltioidantoína , Farmacologia , Análise de Componente Principal , Neoplasias da Próstata , Tratamento Farmacológico , Ligação Proteica , Fisiologia , Conformação Proteica , Receptores Androgênicos , MetabolismoRESUMO
OBJECTIVE@#To research the responsibility and mechanism of auditory event-related potentials (AERP) evoked by pure-tone and different Chinese speech (including single-syllable words, two-syllable words and munbers) stimuli respectively. Meanwhile, to explore feasibility of using different Chinese speech forms as stimulation to evoke AERP in Chinese people.@*METHOD@#AERPs were measured in 37 young normal post-graduated students (70 ears) with pure-tone and three different Chinese speech forms as stimuli. The mean incidence rates, latencies and amplitudes of AERPs were analyzed, the waveforms of AERPs were quantitatively scored. Then all data were statistically analyzed and compared.@*RESULT@#The typical wave P1, N1, P2, N2 and P3 of AERPs could be recorded by both pure-tone and three different Chinese speech stimuli, in which the Chinese two-syllable word was most likely to evoked AERP than pure-tone and remains of Chinese speech forms, and the differences were statistical significance (Chi2 = 0.046, P 0.05) among different stimuli. However, the waveforms of AERPs were smoother and much satisfied evoked by Chinese character and two-syllable word than pure-tone and Chinese number stimuli. The latencies of P3 evoked by number stimulus were significant longer than by other stimuli (P 0.05).@*CONCLUSION@#Besides pure-tone and Chinese single-syllable words, other Chinese speech such as two-syllable words and numbers could be used to evoke AERP with satisfied waveform, which indicated that the Chinese character, two-syllable words and numbers were available to apply for AERP measurement as stimuli for Chinese people.