Improved Gamma-Secretase Inhibitor Screening Assays
AssigneesVib Vzw, Katholieke Universiteit Leuven
InventorsWim Annaert, Ragna Sannerud, Marine Bretou
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Abstract
Current application relates to the field of neurodegenerative diseases. Specifically, the present invention relates to screening methods to identify therapeutic candidates for the prevention and/or treatment of Alzheimer's disease. More particularly, said candidates overcome endolysosomal dysfunction resulting from an accumulation of APP-carboxyterminal fragments.
Claims16
A method of detecting an improved amyloid precursor protein (APP) processing modulator from a collection of gamma-secretase modulators, the method comprising:
- administering a gamma-secretase modulator from the collection to one or more cells expressing a gamma-secretase complex and comprising a polypeptide having at least 95% amino acid identity to SEQ ID No. 1;
- quantifying the level of APP-carboxyterminal fragment (CTF) in the one or more cells before and after administering the gamma-secretase modulator;
- optionally determining the Ca2+ level and/or the cholesterol level in the lysosomes of the one or more cells before and after administering the gamma-secretase modulator;
- detecting that the APP-CTF level is at most 50% increased after the administration of the gamma-secretase modulator compared to before the administration and optionally if the Ca2+ level in the lysosomes is at most 20% decreased and/or the cholesterol level in the lysosomes is at most 25% increased after the administration of the gamma-secretase modulator compared to before the administration.
A method of detecting an improved APP processing modulator, the method comprising:
- providing one or more cells expressing a functional gamma-secretase complex and comprising a polypeptide having at least 95% amino acid identity to SEQ ID No. 1 hereof;
- quantifying amyloid beta (Ab) peptides with a length of 38, 40, 42 and/or 43 amino acids produced in said one or more cells before and after administering a test compound;
- quantifying before and after administering the test compound the level of APP-CTF in the one or more cells and/or the Ca2+ level in the lysosomes of the one or more cells and/or the cholesterol level in the lysosomes of the one or more cells;
- detecting a statistically significant increase in the ratio of Ab38/Ab42, Ab40/Ab42, Ab40/Ab43 or of Ab (38+40)/Ab (42+43) compared to before administering the test compound and at most a 50% increased APP-CTF level and/or at most a 20% decrease in lysosomal Ca2+ level and/or at most a 25% increase lysosomal cholesterol level after the administration of the test compound compared to before administration.
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Dependencies are inferred from the claim text.