Category: Assay Development and Screening

1084-E - High Content Screening of APP processing inhibitors in an ischemic stroke in vitro model: Calcium Channel Blockers (CCB) as neuroprotective compounds

Wednesday, February 7, 2018
11:30 AM - 12:30 PM

There is increasing evidence for influence of Alzheimer´s proteins and neuropathology on ischemic brain injury. Production and deposition of β‐amyloid protein is an important key player involved in mechanism of Alzheimer´s disease as well as ischemic neurodegeneration. To date, efficient therapeutics drugs are still insufficient, making necessary to find relevant compounds amongst masses of potential candidates. A first cell‐based screening model for secretase inhibitors using MCDK cells was performed followed by a second assay in cortical neurons where cell parameters associated with membrane damage, cell viability and neurite outgrowth were measured using High Content Screening (HCS) analysis. In the primary screen we identified 57 inhibitors of Amyloid Precursor Protein (APP) processing in the 1200 compounds of Prestwich library. These initial hits were further analyzed for their capacity to protect primary cortical neurons against the neurotoxicity induced by oxygen glucose deprivation (OGD). In this neuronal screening, just only 4 of these 57 compound (metergoline, bepridil, carbetapentane and raloxifene), all antagonist of calcium channels, displayed as highly neuroprotective effect in a pre‐treatment toxicity assay but only metergoline compound remained active when tested in a post-treatment schedule administered six hours after OGD damage. Our work establishes the feasibility of identifying molecule inhibitors of neuronal damage using both screening models and provides novel insights into the mechanisms of neuroprotection by calcium channels antagonists.

Aida Clarisa Salado

C.E.O.
INNOPROT
Derio-Bizkaia, Pais Vasco, Spain

Dr. Clarisa Salado holds a PhD in Biochemistry and has more than 18 years of experience in fields of pharmacology, drug screening and cancer. She has in the industry since 2002 working in Inbiomed and University of the Basque Country as Researcher and in the Biotechnology Company, Dominion Pharmakine as Biotechnology Division Director. She is currently the CEO of Innoprot.