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Identification of Small Molecule Inhibitors of Tau Aggregation by Targeting Monomeric Tau As a Potential Therapeutic Approach for Tauopathies

View Article: PubMed Central

ABSTRACT

A potential strategy to alleviate the aggregation of intrinsically disordered proteins (IDPs) is to maintain the native functional state of the protein by small molecule binding. However, the targeting of the native state of IDPs by small molecules has been challenging due to their heterogeneous conformational ensembles. To tackle this challenge, we applied a high-throughput chemical microarray surface plasmon resonance imaging screen to detect the binding between small molecules and monomeric full-length Tau, a protein linked with the onset of a range of Tauopathies. The screen identified a novel set of drug-like fragment and lead-like compounds that bound to Tau. We verified that the majority of these hit compounds reduced the aggregation of different Tau constructs in vitro and in N2a cells. These results demonstrate that Tau is a viable receptor of drug-like small molecules. The drug discovery approach that we present can be applied to other IDPs linked to other misfolding diseases such as Alzheimer’s and Parkinson’s diseases.

No MeSH data available.


Related in: MedlinePlus

Comparison of N2a cell viability (MTT assay) with the amount of ThS+ cells (FACS) at increasing compound concentrations. Promising compounds should show a relatively high stable cell viability value over a broad concentration range whereas the amount of ThS+ cells should drop already at low compound concentrations (A). By this criterion, compound ID220149 (B) is the best inhibitor in this series wereas ID220218 (C) and ID220255 (D) show less effectivity and drug-like performance. Note: The MTT value in the untreated sample (without compound) is defined as 100% cell viability. The amount of ThS+ cells in the Tau-induced control is normalized to 100%.
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Figure 6: Comparison of N2a cell viability (MTT assay) with the amount of ThS+ cells (FACS) at increasing compound concentrations. Promising compounds should show a relatively high stable cell viability value over a broad concentration range whereas the amount of ThS+ cells should drop already at low compound concentrations (A). By this criterion, compound ID220149 (B) is the best inhibitor in this series wereas ID220218 (C) and ID220255 (D) show less effectivity and drug-like performance. Note: The MTT value in the untreated sample (without compound) is defined as 100% cell viability. The amount of ThS+ cells in the Tau-induced control is normalized to 100%.

Mentions: We selected three representative synthesized hits, ID220149, ID220218 and ID220255, due to their superior ability to reduce aggregation of the different constructs of Tau in vitro and in N2a cells and for their CNS drug-like physico-chemical properties, and tested them for their ability to inhibit Tau aggregation and cell viability in the inducible N2aTau4RDΔK280 cell line [37] at increasing concentrations (0, 5, 10, 30 and 60 µM). The aggregate load in the N2a cells was quantified via ThS+ cells counted by FACS and the cell viability was measured by MTT-assay (Fig. 6).


Identification of Small Molecule Inhibitors of Tau Aggregation by Targeting Monomeric Tau As a Potential Therapeutic Approach for Tauopathies
Comparison of N2a cell viability (MTT assay) with the amount of ThS+ cells (FACS) at increasing compound concentrations. Promising compounds should show a relatively high stable cell viability value over a broad concentration range whereas the amount of ThS+ cells should drop already at low compound concentrations (A). By this criterion, compound ID220149 (B) is the best inhibitor in this series wereas ID220218 (C) and ID220255 (D) show less effectivity and drug-like performance. Note: The MTT value in the untreated sample (without compound) is defined as 100% cell viability. The amount of ThS+ cells in the Tau-induced control is normalized to 100%.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4997948&req=5

Figure 6: Comparison of N2a cell viability (MTT assay) with the amount of ThS+ cells (FACS) at increasing compound concentrations. Promising compounds should show a relatively high stable cell viability value over a broad concentration range whereas the amount of ThS+ cells should drop already at low compound concentrations (A). By this criterion, compound ID220149 (B) is the best inhibitor in this series wereas ID220218 (C) and ID220255 (D) show less effectivity and drug-like performance. Note: The MTT value in the untreated sample (without compound) is defined as 100% cell viability. The amount of ThS+ cells in the Tau-induced control is normalized to 100%.
Mentions: We selected three representative synthesized hits, ID220149, ID220218 and ID220255, due to their superior ability to reduce aggregation of the different constructs of Tau in vitro and in N2a cells and for their CNS drug-like physico-chemical properties, and tested them for their ability to inhibit Tau aggregation and cell viability in the inducible N2aTau4RDΔK280 cell line [37] at increasing concentrations (0, 5, 10, 30 and 60 µM). The aggregate load in the N2a cells was quantified via ThS+ cells counted by FACS and the cell viability was measured by MTT-assay (Fig. 6).

View Article: PubMed Central

ABSTRACT

A potential strategy to alleviate the aggregation of intrinsically disordered proteins (IDPs) is to maintain the native functional state of the protein by small molecule binding. However, the targeting of the native state of IDPs by small molecules has been challenging due to their heterogeneous conformational ensembles. To tackle this challenge, we applied a high-throughput chemical microarray surface plasmon resonance imaging screen to detect the binding between small molecules and monomeric full-length Tau, a protein linked with the onset of a range of Tauopathies. The screen identified a novel set of drug-like fragment and lead-like compounds that bound to Tau. We verified that the majority of these hit compounds reduced the aggregation of different Tau constructs in vitro and in N2a cells. These results demonstrate that Tau is a viable receptor of drug-like small molecules. The drug discovery approach that we present can be applied to other IDPs linked to other misfolding diseases such as Alzheimer’s and Parkinson’s diseases.

No MeSH data available.


Related in: MedlinePlus