Appendix Table 1. .
Comparison of the Concentration-Dependent Generation of H2O2 by Compounds in Different Reducing Agents
|
Compounds |
DTT |
TCEP |
GSH |
HBSS |
||||
|---|---|---|---|---|---|---|---|---|
| PubChem-SID | % Activation at 50 μM | AC50 μM | % Activation at 50 μM | AC50 μM | % Activation at 50 μM | AC50 μM | % Activation at 50 μM | AC50 μM |
| 14727570 | 134.74 | <0.01 | 171.40 | 25.30 | 3.60 | >50 | 0.80 | >50 |
| 14727734 | 115.48 | 0.19 | 3.99 | >50 | −0.60 | >50 | 1.80 | >50 |
| 22404392 | 128.95 | 0.43 | 53.25 | 48.70 | 3.30 | >50 | 1.00 | >50 |
| 16952991 | 109.74 | 0.72 | 1.70 | >50 | 2.50 | >50 | 0.50 | >50 |
| 14723875 | 111.50 | 0.90 | 86.65 | 23.10 | 2.50 | >50 | 6.70 | >50 |
| 845167 | 112.80 | 0.90 | 57.30 | 37.90 | 2.40 | >50 | 1.80 | >50 |
| 11532976 | 121.55 | 1.48 | 157.10 | 4.10 | 2.70 | >50 | 1.40 | >50 |
| 24809205 | 110.69 | 1.51 | 68.60 | 25.50 | 4.30 | >50 | 1.50 | >50 |
| 24814503 | 127.08 | 2.73 | 19.80 | >50 | 1.10 | >50 | 1.10 | >50 |
| 14726526 | 114.91 | 2.76 | 91.89 | 6.80 | 2.90 | >50 | 2.50 | >50 |
| 14730721 | 114.50 | 2.88 | 0.00 | >50 | 1.40 | >50 | 0.70 | >50 |
| 24809506 | 116.49 | 3.11 | 16.60 | >50 | 3.00 | >50 | 1.50 | >50 |
| 14742005 | 78.78 | 6.44 | 0.10 | >50 | 3.10 | >50 | 0.60 | >50 |
| 847359 | 133.23 | 7.85 | 110.00 | >50 | 1.40 | >50 | 0.90 | >50 |
| 17507336 | 123.59 | 9.42 | 5.70 | >50 | 0.40 | >50 | 0.10 | >50 |
| 14719117 | 107.90 | 9.85 | 0.20 | >50 | 3.50 | >50 | 0.80 | >50 |
| 17508465 | 115.53 | 11.77 | 4.00 | 28.00 | 3.40 | >50 | 0.80 | >50 |
| 17511877 | 117.92 | 12.27 | 0.20 | >50 | 3.30 | >50 | 0.70 | >50 |
| 17510947 | 126.66 | 12.50 | 23.00 | >50 | 0.70 | >50 | 0.30 | >50 |
| 22405508 | 131.06 | 12.84 | 120.93 | 13.30 | 2.10 | >50 | 16.20 | >50 |
| 24812833 | 110.79 | 12.87 | 7.90 | >50 | 3.60 | >50 | 1.40 | >50 |
| 17510616 | 121.21 | 12.98 | 29.20 | >50 | 0.80 | >50 | 1.20 | >50 |
| 14727791 | 94.83 | 18.44 | 0.50 | >50 | 3.10 | >50 | 0.30 | >50 |
| 14742458 | 129.86 | 19.15 | 2.20 | >50 | 2.70 | >50 | 0.50 | >50 |
| 17510946 | 114.68 | 23.26 | 55.59 | 45.50 | 1.40 | >50 | 2.20 | >50 |
| 24813200 | 112.06 | 24.03 | 21.00 | >50 | −0.20 | >50 | 0.00 | >50 |
| 14742370 | 122.00 | 26.04 | 1.20 | >50 | 3.10 | >50 | 0.30 | >50 |
| 22400632 | 126.92 | 28.27 | −1.70 | >50 | 3.60 | >50 | 1.10 | >50 |
| 857446 | 127.03 | 28.18 | 1.00 | >50 | 3.40 | >50 | 1.20 | >50 |
| 17433753 | 113.55 | 34.04 | 24.60 | >50 | 4.00 | >50 | 0.80 | >50 |
| 22412622 | 94.24 | 38.24 | 32.90 | >50 | 4.10 | >50 | 2.50 | >50 |
| 17516177 | 71.07 | 39.24 | 0.50 | >50 | 3.60 | >50 | 1.30 | >50 |
| 17513322 | 121.83 | 40.61 | 15.90 | >50 | 6.80 | >50 | 1.70 | >50 |
| 17504835 | 90.21 | 40.90 | 0.10 | >50 | 0.40 | >50 | 0.10 | >50 |
| 17505825 | 98.90 | 41.24 | 0.10 | >50 | 3.80 | >50 | 1.60 | >50 |
| 24825938 | 64.55 | 45.23 | 0.40 | >50 | 3.60 | >50 | 1.30 | >50 |
| 14742003 | 62.96 | 48.41 | 1.40 | >50 | 3.20 | >50 | 0.70 | >50 |
The amount of H2O2 generated by compounds in the presence of reducing agents was expressed as the % activation of the mean maximum (100 μM H2O2, n = 32) and mean minimum (0.5% DMSO in HBSS, n = 24) plate controls calculated with an ActivityBase™ concentration–response template. The % activation at the highest compound concentration tested (50 μM) is presented together with the AC50 (50% effective concentration of activation in μM) calculated using an XLfit 4-parameter logistic (sigmoidal dose–response) model: y = (A+((B-A)/(1+((C/x)ˆD)))), where y is the percent activation and x is the corresponding compound concentration. The fitted C parameter is the AC50 and defined as the concentration giving a response half way between the fitted top (B) and bottom (A) of the curve. The A and B parameters were locked as 0 and 100, respectively. The 10-point AC50 assays were conducted in single wells per concentration, and performed in Hank’s balanced salt solution (HBSS) in the absence or presence of a 0.8 mM concentration of 3 different reducing agents; dithiothreitol (DTT); reduced glutathione (GSH); or TCEP: tris(2-carboxyethyl)phosphine. Compounds were identified by their PubChem substance identifier (SID) numbers.
Abbreviations: DTT, Dithiothreitol; SID, PubChem Substance Identity Number.