Table 1.
Compound potency in kinase and cell-based assays
| Compound name | IC50 WT (nM) (n = ) | IC50 T618Q (nM) (n = ) | EC50 WT (nM) (n = ) | EC50 T618Q (nM) (n = ) | EC90 WT (nM) (n = ) | EC90 T618Q (nM) (n = ) |
|---|---|---|---|---|---|---|
| ML1 | 3.1 + /−0.22 (68) | 8440 + /−1070 (14) | 395.0 + /−21.9 (82) | 5952 + /−594.7 (14) | 696.8 + /−98 .9 (82) | 20,715 + /−3209 (14) |
| ML2 | 1.71 + /−0.22 (2) | 1713 + /−218.5 (2) | 196.6 + /−13.4 (6) | 140.2 + /−12.0 (3) | 627.6 + /−125.8 (6) | 955.7 + /−58.2 (3) |
| ML3 | 0.79 + /−0.15 (2) | 842 + /−74.9 (2) | 63.4 + /−3.7 (9) | 195.1 + /−8.1 (3) | 235.5 + /−18.7 (9) | 278.7 + /−29.4 (3) |
| ML4 | 0.45 + /−0.04 (2) | 231 + /−9.1 (2) | 25.3 + /−1.7 (12) | 106.7 + /−12.2 (3) | 106.6 + /−13.4 (12) | 185.8 + /−26.2 (3) |
| ML5 | 0.88 + /−0.04 (2) | >100,000 (4) | 329.1 + /−3.5 (6) | 12,735 + /−1519 (3) | 445.2 + /−2.5 (6) | 61,465 + /−1542 (3) |
| ML6 | 0.13 + /−0.02 (2) | 52,500 + /−1000 (2) | 102.3 + /−13.4 (6) | 1855 + /−147.5 (3) | 141.5 + /−12.2 (6) | 3491 + /−822 (3) |
| ML7 | 4.34 + /−0.04 (2) | >100,000 (2) | 488.9 + /−39.4 (6) | 35,111 + /−2739 (3) | 633.3 + /−15.8 (6) | 60,663 + /−2931 (3) |
| ML8 | 0.79 + /−0.01 (2) | 73,730 (4) | 148.1 + /−10.6 (9) | 5574 + /−474.1 (3) | 181.6 + /−8.6 (9) | 31,022 + /−2243 (3) |
| ML9 | 1.19 + /−0.07 (2) | >100,000 (2) | 104.6 + /−5.3 (6) | 6900 + /−922.1 (3) | 158.1 + /−6.8 (6) | 48,327 + /−1790 (3) |
| ML10 | 0.16 + /−0.01 (2) | 29,540 + /−519 (2) | 2.1 + /−0.2 (18) | 2430 + /−413.3 (6) | 4.50 + /−0.5 (18) | 4612 + /−1549 (6) |
Compounds were tested for their ability to inhibit the protein kinase activity of the full length 6His-tagged PfPKG recombinant protein (IC50). This was carried out using a microfluidic mobility shift assay, which measures the separation of phosphorylated/non-phosphorylated forms of a fluorescent peptide substrate to evaluate conversion. Hypoxanthine incorporation assays were then used to measure the ability of the newly synthesized compounds to inhibit P. falciparum (3D7 and T618Q clonal lines) asexual blood stage growth in vitro (EC50 and EC90). Data are in nM ( + /− the s.e.m.). The number of biological replicates (carried out in triplicate) is shown in brackets