TABLE 4.
kapp values of selected alkyne and desthiobiotin probes against a panel of trypsin-like serine proteases and the description of the mechanism of inhibition defined by the progress curves: slow-binding (Rev.) or irreversible (Irrev.).
| ||||||||
|---|---|---|---|---|---|---|---|---|
| # | R1 | R2 | Alk/DtB a | Trypsin-3 | β-tryptase | Thrombin | uPA b | CatG b |
| kapp (M−1 s−1) c | ||||||||
| 32b |
|
H | Alk | 18 × 102 ± 66 Irrev. | 41 ± 5 Irrev. | 64 ± 8 Rev. | 41 ± 4 Irrev. | * d Rev. |
| 45b | DtB | 17 × 102 ± 122 Irrev. | 237 ± 42 Irrev. | 19 ± 2 Rev. | N.D. | N.D. | ||
| 33b | Cl | Alk | 35 × 102 ± 40 Irrev. | 28 × 102 ± 82 Irrev. | 11 × 102 ± 42 Rev. | 15 × 102 ± 12 Irrev. | * d Rev. | |
| 46b | DtB | 92 × 102 ± 349 Irrev. | 75 × 102 ± 1475 Irrev. | 63 × 102 ± 290 Rev. | 940 ± 37 Irrev. | N.D. | ||
| 34b |
|
H | Alk | 40 × 103 ± 2642 Rev. | 712 ± 41 Rev. | 17 ± 1 Rev. | 75 × 102 ± 725 Irrev. | * d Rev. |
| 50b | DtB | 49 × 103 ± 2979 Rev. | 32 × 102 ± 613 Rev. | 47 ± 5 Rev. | 33 × 103 ± 12 × 103 Irrev. | * d Rev. | ||
| 35b | NHCOCH3 | Alk | * d Rev. | 70 × 102 ± 582 Rev. | 128 ± 5 Rev. | 13 × 104 ± 30 × 103 Irrev. | 477 ± 67 Rev. | |
| 52b | DtB | 86 × 103 ± 8250 Rev. | 18 × 103 ± 1184 Rev. | 49 ± 5 Rev. | 38 × 103 ± 4175 Irrev. | * d Rev. | ||
| 36b | Cl | Alk | 29 × 103 ± 6499 Rev. | 61 × 103 ± 6860 Rev. | 251 ± 16 Rev. | 32 × 104 ± 29 × 103 Irrev. | * d Rev. | |
| 54b | DtB | * d Rev. | * d Rev. | * d Rev. | 23 × 104 ± 36 × 103 Irrev. | * d Rev. | ||
| 37b |
|
H | Alk | 293 ± 34 Irrev. | 375 ± 34 Irrev. | N.D. | 124 ± 23 Irrev. | 582 ± 2 Irrev. |
| 55b | DtB | 231 ± 18 Irrev. | 347 ± 19 Irrev. | N.D. | 77 ± 1 Irrev. | 301 ± 85 Irrev. | ||
| 38b | NHCOCH3 | Alk | 358 ± 6 Irrev. | 20 × 102 ± 54 Irrev. | N.D. | 215 ± 7 Irrev. | 14 × 102 ± 52 Irrev. | |
| 56b | DtB | 334 ± 85 Irrev. | 321 ± 2 Irrev. | N.D. | 62 ± 2 Irrev. | 420 ± 37 Irrev. | ||
| 39b | Cl | Alk | 11 × 102 ± 164 Irrev. | 45 × 102 ± 547 Irrev. | N.D. | 25 × 102 ± 86 Irrev. | 23 × 102 ± 347 Rev. | |
| 57b | DtB | 13 × 102 ± 340 Irrev. | 65 × 102 ± 1,203 Irrev. | N.D. | 25 × 102 ± 11 Irrev. | 11 × 102 ± 117 Rev. | ||
| 41b |
|
H | Alk | 17 × 102 ± 142 Irrev. | 27 ± 2 Irrev. | 41 × 102 ± 38 Irrev. | N.D. | N.D. |
| 31b | Pro-Lys | H | Alk | 13 × 102 ± 28 Irrev. | 136 ± 3 Irrev. | 13 × 102 ± 5 Irrev. | 188 ± 1 Irrev. | N.D. |
Alk, alkyne probe; DtB, desthiobiotin probe.
Panel of serine proteases abbreviations: uPA, urokinase plasminogen activator; catG, cathepsin G.
kapp are calculated from two independent experiments; when SD was higher than three times the average value, a third independent experiment was run (mean ± SD).
kapp determination is not possible by curve fitting.
N.D.: IC50 is greater than 10 μM, and progress curves were not performed.