Table 1.
Hit compound structures and corresponding assay activities from the L. donovani and T. cruzi screening campaigns.
| Activitya | Compound Id. | Structure | IDb and priorityc |
L. donovani |
T. cruzi |
HepG2e (pCC50) | |||
|---|---|---|---|---|---|---|---|---|---|
| Axenic assay (pIC50) | InMac assay (pIC50) | InMac HS assayd (pIC50) | High-content imaging assay (pIC50) | Trypomastigote assay (pIC50) | |||||
| Both parasites | TCOLFS068570 |  |
Cluster A (H) | 5.93 ± 0.01 | AmMacf 6.3 ± 0.25 InfCelg 6.27 ± 0.2 Mach <4.3 |
AmMac HS 6.17 ± 0.19 InfCel HS 6.09 ± 0.11 Mac HS 4.53 ± 0.28 |
AmCel 5.85 ± 0.11 InfCel 4.4 ± 0.25 H9c2i < 4.3 |
24 h < 4.3 48 h < 4.3 72 h < 4.3 |
<4 |
| TCOLFS018919 |  |
Cluster B (H) | 5.80 ± 0.15 | AmMac 5.68 ± 0.30 InfCel 5.09 ± 0.65 Mac <4.3 |
AmMac HS 4.89 ± 0.32 InfCel HS 4.75 ± 0.30 Mac HS < 4.3 |
AmCel 5.71 ± 0.30 InfCel 5.38 ± 0.30 H9c2 <4.3 |
24 h < 4.3 48 h < 4.3 72 h 4.53 ± 0.27 |
<4 | |
| TCOLFS026832 |  |
Cluster C (H) | 6.53 ± 0.03 | AmMac 6.47 ± 0.26 InfCel 6.32 ± 0.17 Mac 4.5 ± 0.31 |
AmMac HS 6.13 ± 0.05 InfCel HS 5.96 ± 0.09 Mac HS 4.62 ± 0.45 |
AmCel 7.98 ± 0.23 InfCel 7.64 ± 0.28 H9c2 <4.3 |
24 h 7.28 48 h 7.08 72 h 7.41 ± 0.19 |
<4 | |
| TCOLFS026398 |  |
Cluster D (H) | - - | AmMac 5.09 ± 0.06 InfCel 5.03 ± 0.1 Mac <4.3 |
- - | AmCel 6.65 ± 0.2 InfCel 6.37 ± 0.2 H9c2 <4.3 |
24 h to −48 h to −72 h 6.06 ± 0.16 | <4 | |
| TCOLFS129266 |  |
Cluster E (L) | 6.54 ± 0.01 | AmMac 6.28 ± 0.33 InfCel 5.62 ± 0.78 Mac 4.90 ± 0.18 |
- - | AmCel 6.77 ± 0.01 InfCel 6.68 ± 0.03 H9c2 4.42 ± 0.02 |
24 h to −48 h to −72 h 6.24 ± 0.01 | 4.69 ± 0.01 | |
| TCOLFS098882 |  |
Singleton (H) | 5 ± 0.04 | AmMac 5.82 ± 0.55 InfCel 5.2 ± 0.09 Mac 4.46 ± 0.22 |
- - | AmCel 5.37 ± 0.06 InfCel 5.17 ± 0.24 H9c2 5.61 |
24 h to −48 h to −72 h 5.66 ± 0.01 | <4 | |
| TCOLFS006487 |  |
Singleton (L) | 5.23 ± 0.05 | AmMac 5.64 ± 0.32 InfCel 5.38 ± 0.09 Mac 4.53 ± 0.25 |
- - | AmCel 5.89 ± 0.06 InfCel 5.79 ± 0.06 H9c2 5.28 ± 0.1 |
24 h to −48 h to −72 h 5.94 ± 0.02 | 4.33 ± 0.38 | |
| L. donovani | TCOLFS079469 |  |
Singleton (H) | 6.38 ± 0.03 | AmMac 6.25 ± 0.28 InfCel 4.62 ± 0.46 Mac <4.3 |
- - | AmCel <4.3 InfCel <4.3 H9c2 <4.3 |
24 h to −48 h to −72 h < 4.3 | <4 |
| TCOLFS124301 |  |
Singleton (H) | 5.51 | AmMac 5.46 ± 0.04 InfCel 5.28 ± 0.08 Mac <4.3 |
- - | - - | - - | 4.58 ± 0.82 | |
| T. cruzi | TCOLFS008553 |  |
Cluster F (H) | - - | AmMac <4.3 InfCel <4.3 Mac 4.42 ± 0.19 |
AmMac HS < 4.3 InfCel HS < 4.3 Mac HS 4.52 ± 0.27 |
AmCel 6.3 ± 0.11 InfCel 5.88 ± 0.11 H9c2 4.36 ± 0.08 |
24 h 5.66 ± 0.06 48 h 5.31 ± 0.08 72 h 5.44 ± 0.05 |
<4 |
| TCOLFS129639 |  |
Cluster G (L) | - - | AmMac 5.01 ± 0.17 InfCel 4.31 ± 0.01 Mac 4.74 ± 0.21 |
AmMac HS 4.78 ± 0.15 InfCel HS 4.52 ± 0.06 Mac HS 4.42 ± 0.16 |
AmCel 6.45 ± 0.11 InfCel 6.13 ± 0.27 H9c2 4.41 ± 0.12 |
24 h 4.8 48 h 5.27 72 h 5.84 ± 0.08 |
<4 | |
| TCOLFS025993 |  |
Cluster H (M) | - - | AmMac 4.84 ± 0.21 InfCel 4.52 ± 0.24 Mac 4.36 ± 0.09 |
AmMac HS 4.64 ± 0.43 InfCel HS < 4.3 Mac HS 4.44 ± 0.28 |
AmCel 6.68 ± 0.2 InfCel 6.49 ± 0.19 H9c2 <4.3 |
24 h < 4.3 48 h < 4.3 72 h 5.59 ± 0.11 |
<4 | |
| TCOLFS059386 |  |
Singleton (L) | - - | - - | - - | AmCel 5.9 ± 0.06 InfCel 5.82 ± 0.1 H9c2 <4.3 |
24 h to −48 h to −72 h 5.27 ± 0.01 | <4 | |
| TCOLFS089113 |  |
Singleton (L) | - - | AmMac <4.3 InfCel <4.3 Mac <4.3 |
AmMac HS 4.73 ± 0.01 InfCel HS 4.41 ± 0.16 Mac HS 4.89 ± 0.04 |
AmCel 6.3 ± 0.12 InfCel 6.03 ± 0.18 H9c2 <4.3 |
24 h < 4.3 48 h < 4.3 72 h 4.83 ± 0.77 |
<4 | |
| TCOLFS135869 |  |
Cluster I (L) | - - | - - | - - | AmCel 6.85 ± 0.14 InfCel 6.48 ± 0.1 H9c2 <4.3 |
24 h to −48 h to −72 h 6.33 ± 0.06 | <4 | |
| TCOLFS112845 |  |
Singleton (M) | - - | - - | - - | AmCel 6.07 ± 0.04 InfCel 5.98 ± 0.07 H9c2 <4.3 |
24 h to −48 h to −72 h 5.27 ± 0.01 | <4 | |
| TCOLFS002713 |  |
Singleton (M) | - - | - - | - - | AmCel 6.27 ± 0.01 InfCel 5.97 ± 0.08 H9c2 <4.3 |
24 h to −48 h to −72 h 5.72 ± 0.01 | <4 | |
| TCOLFS050529 |  |
Singleton (L) | - - | - - | - - | AmCel 6.0 ± 0.01 InfCel 5.88 ± 0.08 H9c2 <4.3 |
24 h to −48 h to −72 h 4.42 ± 0.1 | <4 | |
| TCOLFS099080 |  |
Singleton (L) | - - | - - | - - | AmCel 6.35 ± 0.02 InfCel 5.87 ± 0.11 H9c2 <4.3 |
24 h to −48 h to −72 h 5.64 ± 0.01 | 4.73 | |
| TCOLFS013976 |  |
Singleton (L) | - - | - - | - - | AmCel 6.13 ± 0.03 InfCel 5.76 ± 0.1 H9c2 <4.3 |
24 h to −48 h to −72 h 5.69 ± 0.01 | <4 | |
| TCOLFS148231 |  |
Singleton (M) | - - | - - | - - | AmCel 6.21 InfCel 5.87 ± 0.11 H9c2 4.85 ± 0.05 |
24 h to −48 h to −72 h 5.64 | <4 | |
- - not tested.
*Compounds determined to not be novel; novel compounds shown in Table 3.
Compounds are classified per their antiparasitic activity (both parasites, L. donovani, or T. cruzi).
Hits are categorized as either a hit cluster (2 or more related analogs; clusters A−I) or singletons (i.e., those hits with no related analogs).
The hit priority level based on biological activity (H: high, meets desirable activity profile; M: medium, slow-acting compound or missing activity profiling; L: low, undesirable activity profile).
HS denotes replacement of FBS with horse serum in the L. donovani inMac assay.
Additional mammalian cytotoxicity was measured against HepG2 (hepatocellular carcinoma) cell line.
AmCel: High-content imaging assay out measuring the number of amastigotes per cell.
InfCel: High-content imaging assay out measuring the number of infected host cells per well.
Mac: Macrophage host cell for L. donovani intracellular high-content imaging assay.
H9c2: Embryonic cardiomyocyte host cell for T. cruzi intracellular high-content imaging assay.