Table 1.
Assays of the inhibition of hDAGLα (radio-TLC assay with [14C]SAG substrate),37 rFAAH,57 and hMAGL63 enzyme activities
| Compound Number | Detail of N-formyl- α-amino ester | hDAGLα inhibition at 10 μM detergent free or (with Triton X-100) (%) | rFAAH inhibition at 10 μM (%) | hMAGL inhibition at 10 μM (%) |
|---|---|---|---|---|
| 3 (THL) | L-leucyl | 100, (100A) | 6 | 47 |
| 4 (OMDM-188) | L-isoleucyl | (98A) | 7 | 16 |
| 5 | L-allo-isoleucyl | (95A) | 3 | 1 |
| 6 | D-isoleucylC | (78A) | 28 | 39 |
| 7 | D-allo-isoleucyl | (86A) | 0 | 42 |
| 8 | (S)-α-aminobutyryl | (99A) | 12 | 45 |
| 9 | none | (25A, 45B) | 10 | 4 |
| trans-10 | none | (37B) | 79 | 55 |
| cis-11 | none | (25B) | 94D | 100D |
| JZL184 | NA | (37B) | 97E | 100F |
| URB597 | NA | ND | 100E | 18 |
| n-C16H33SO2F | NA | ND | 100E | 82 |
| PMSF | NA | 19 | 100E | 5 |
| RHC80267 (1) | NA | 30 | 95E | 22 |
| SD41 | NA | <0* | 10 | 13 |
NA, not applicable; ND, not done
A
0.05% Triton X-100 present
B
0.015% Triton X-100 present
C
5% impurity in D-isoleucyl analog due to D-allo-isoleucyl analog
D
cis-11 inhibits rFAAH only 13% at 1 μM, but inhibits hMAGL 66% at 100 nM
E
8 pt rFAAH IC50 (95% confidence)
| JZL184 | 974 nM (784–1210) |
| URB597 | 4.9 nM (4.1–6.0) |
| n-C16H33SO2F | 6.3 nM (4.5–8.7) |
| PMSF | 833 nM (746–931) |
| RHC80267 (1) | 2,240 nM (2010–2500) |
F
8 pt hMAGL IC50 (95% confidence)
| JZL184 | 57 nM (53–62) |
*
protein was pretreated with JZL184 to completely inhibit MAGL activity