Novel compounds lowering the cellular isoform of the human prion protein in cultured human cells |
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| Institution: | 1. Institute for Neurodegenerative Diseases, University of California, San Francisco, United States;2. Department of Neurology, University of California, San Francisco, United States;3. Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, United States;4. Small Molecule Discovery Center, University of California, San Francisco, United States;5. Department of Pharmaceutical Chemistry, University of California, San Francisco, United States;1. Collaborations in Chemistry, 5616 Hilltop Needmore Road, Fuquay-Varina, NC 27526, USA;2. Collaborative Drug Discovery, 1633 Bayshore Highway, Suite 342, Burlingame, CA 94010, USA;3. Center for Tuberculosis Research, Johns Hopkins University School of Medicine, Baltimore, MD, USA;4. Department of Pharmacology & Physiology, Rutgers University – New Jersey Medical School, 185 South Orange Avenue, Newark, NJ 07103, USA;5. Department of Medicine, Center for Emerging and Reemerging Pathogens, Rutgers University – New Jersey Medical School, 185 South Orange Avenue, Newark, NJ 07103, USA;1. Department of Medicinal Chemistry, Faculty of Pharmacy and Drug Design & Development Research Center, Tehran University of Medical Sciences, Tehran 14176, Iran;2. Pharmacology and Toxicology Department, Pharmaceutical Sciences Branch and Pharmaceutical Sciences Research Center, Islamic Azad University, Tehran, Iran;3. Department of Radiopharmacy, School of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran;4. Department of Pharmaceutical Chemistry, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran;5. Department of Physiology & Pharmacology, School of Medicine, Kashan University of Medical Sciences, Kashan, Iran;6. Endocrinology and Metabolism Research Center, Tehran University of Medical Sciences, Tehran, Iran;1. Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore 560012, India;2. LigDCipher, 729 Nevada Ave, San Mateo, CA 94402, USA;3. Collaborative Drug Discovery, 1633 Bayshore Highway, Suite 342, Burlingame, CA 94403, USA;4. Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore 560064, India;1. Cellular Biochemistry Laboratory, Methodist Research Institute, Indiana University Health, 1800 N. Capitol Ave., Room E504, Indianapolis, IN 46202, USA;2. Department of Medicine, Indiana University School of Medicine, Indianapolis, IN 46202, USA;3. Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University, 575 Stadium Mall Dr, West Lafayette, IN 47907, USA |
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| Abstract: | Purpose: Previous studies showed that lowering PrPC concomitantly reduced PrPSc in the brains of mice inoculated with prions. We aimed to develop assays that measure PrPC on the surface of human T98G glioblastoma and IMR32 neuroblastoma cells. Using these assays, we sought to identify chemical hits, confirmed hits, and scaffolds that potently lowered PrPC levels in human brains cells, without lethality, and that could achieve drug concentrations in the brain after oral or intraperitoneal dosing in mice. Methods: We utilized HTS ELISA assays to identify small molecules that lower PrPC levels by ?30% on the cell surface of human glioblastoma (T98G) and neuroblastoma (IMR32) cells. Results: From 44,578 diverse chemical compounds tested, 138 hits were identified by single point confirmation (SPC) representing 7 chemical scaffolds in T98G cells, and 114 SPC hits representing 6 scaffolds found in IMR32 cells. When the confirmed SPC hits were combined with structurally related analogs, >300 compounds (representing 6 distinct chemical scaffolds) were tested for dose–response (EC50) in both cell lines, only studies in T98G cells identified compounds that reduced PrPC without killing the cells. EC50 values from 32 hits ranged from 65 nM to 4.1 μM. Twenty-eight were evaluated in vivo in pharmacokinetic studies after a single 10 mg/kg oral or intraperitoneal dose in mice. Our results showed brain concentrations as high as 16.2 μM, but only after intraperitoneal dosing. Conclusions: Our studies identified leads for future studies to determine which compounds might lower PrPC levels in rodent brain, and provide the basis of a therapeutic for fatal disorders caused by PrP prions. |
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| Keywords: | Prion Neurodegeneration Creutzfeldt–Jakob disease |
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