Antiprion compounds that reduce PrPSc levels in dividing and stationary-phase cells |
| |
Authors: | B. Michael Silber Joel R. Gever Zhe Li Alejandra Gallardo-Godoy Adam R. Renslo Kartika Widjaja John J. Irwin Satish Rao Matthew P. Jacobson Sina Ghaemmaghami Stanley B. Prusiner |
| |
Affiliation: | 1. Institute for Neurodegenerative Diseases, University of California, San Francisco, CA 94143, United States;2. Department of Neurology, University of California, San Francisco, CA 94143, United States;3. Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, CA 94143, United States;4. Small Molecule Discovery Center, University of California, San Francisco, CA 94143, United States;5. Department of Pharmaceutical Chemistry, University of California, San Francisco, CA 94143, United States |
| |
Abstract: | During prion diseases, a normally benign, host protein, denoted PrPC, undergoes alternative folding into the aberrant isoform, PrPSc. We used ELISA to identify and confirm hits in order to develop leads that reduce PrPSc in prion-infected dividing and stationary-phase mouse neuroblastoma (ScN2a-cl3) cells. We tested 52,830 diverse small molecules in dividing cells and 49,430 in stationary-phase cells. This led to 3100 HTS and 970 single point confirmed (SPC) hits in dividing cells, 331 HTS and 55 confirmed SPC hits in stationary-phase cells as well as 36 confirmed SPC hits active in both. Fourteen chemical leads were identified from confirmed SPC hits in dividing cells and three in stationary-phase cells. From more than 682 compounds tested in concentration–effect relationships in dividing cells to determine potency (EC50), 102 had EC50 values between 1 and 10 μM and 50 had EC50 values of <1 μM; none affected cell viability. We observed an excellent correlation between EC50 values determined by ELISA and Western immunoblotting for 28 representative compounds in dividing cells (R2 = 0.75; p <0.0001). Of the 55 confirmed SPC hits in stationary-phase cells, 23 were piperazine, indole, or urea leads. The EC50 values of one indole in stationary-phase and dividing ScN2a-cl3 cells were 7.5 and 1.6 μM, respectively. Unexpectedly, the number of hits in stationary-phase cells was ~10% of that in dividing cells. The explanation for this difference remains to be determined. |
| |
Keywords: | Antiprion compounds Dividing and stationary-phase brain cells |
本文献已被 ScienceDirect 等数据库收录! |
|