Screen Of Small Lead Compounds Targeting Protein-Protein Interaction

Protein–protein interactions (PPI) have a pivotal role in growth, differentiation, the terminal event of programmed cell death and other primary biological processes. PPI is also closely related to the occurrence and development of diseases. Therefore, the intervention of their function can generate novel therapeutic agents. Due to the discovery of protein"hot spots"which are compact regions at many protein–protein interfaces that are crucial for the high affinity of the interaction, many researches focus on the drugs targeting protein-protein interactions.Nrf2 is a transcription factor which is related with oxidative stresses. The inducers of the Keap1-Nrf2-ARE pathway have the function of chemoprevention. These inducers can protect cells and tissues from endogenous and exogenous stresses. In the present study, a high throughput screen model to screen regulative molecules of Keap1-Nrf2-ARE pathway is constructed which is based on the ARE reporter activity. 188 compounds are screened, and the most potent inducer is compound c7. c7 can activate the ARE reporter activity and upregulate the anti-oxidative target genes such as HO-1 and NQO1. Furthermore, c7 can protect L02 cells from the cell death induced by H2O2. We also investigate that c7 induces the ARE pathway by stabilizing the Nrf2 protein and the p38 MAPK pathway is involved in the activation of Keap1-Nrf2-ARE pathway by c7.Another model to screen small molecules for inhibitors of p53-MDM2 protein-protein interaction is established. The system is based on the mammalian two-hybrid technology. 188 compounds are also screened.