Table 4. Summary of the effects of selected synthesized hits on Tau aggregation in vitro and in N2a cells. (Column 5 – 7) The relative ThS-fluorescence was used to indicate the degree of tau fibril formation after compound treatment (note: 2% DMSO were added to the controls to ensure comparable conditions to the compound treated samples). (Column 8) The amount of ThS+ cells (cells with tau aggregates stained by ThS and quantified by FACS) in the induced compound-untreated sample (”DMSO + Dox”) was normalized to 100%. (Column 9) The cell viability (monitored by MTT-assay) of the uninduced compound-untreated sample (entry #1, right column) was normalized to 100%. MTT-values reflect the viability of the N2a cells in the presence of the compounds without expression of tau protein. (Column 10) Concentrations at which synthesized hits were soluble and no compound aggregates were detected by DLS.
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 |
|---|---|---|---|---|---|---|---|---|---|
| Hit ID | Hit Series | MW | Affinity HT-CM-SPRa | hTau2N4Rwtb | Tau3RD c | Tau4RD DK280d |
Tau4RD DK280 N2a cellse |
viability N2a cells (MTT) |
solubility (µM) |
| bb14[34] | 452.0 | 54.3 ± 8.3 | 14.0 ± 0.1 | 42.0 ± 1.6 | |||||
| BSc3094[35] | 380.4 | 7.2 | 101.9 ± 3.8 | ||||||
| DMSO (- Dox) |
100 ± 5.5 | 100 ± 13.0 | 100 ± 5.4 | 0.1 | 100 ± 1.3 | ||||
| DMSO (+ Dox) |
100 | ||||||||
| ID220149 | 7 | 375.6 | medium | 105.8±13.6 | 83.6± 2.6 | 69.8 ± 4.3 | 29.6 | 74.1 ± 0.8 | 300.0 |
| ID220218 | 3 | 359.4 | weak | 97.4± 7.1 | 86.0± 4.7 | 77.1 ± 5.9 | 38.6 | 57.8 ± 3.0 | 300.0 |
| ID220255 | 12 | 406.4 | weak | 113.2 ± 4.4 | 107.9± 2.5 | 78.9 ± 5.4 | 25.0 | 86.6 ±6.5 | 150.0 |
aRelative affinity of immobilized hits to hTau2N4Rwt detected by SPR in the HT-CM-SPR.