| . | IC50 TYK2 (nM) . | IC50 JAK1 (nM) (TYK2 > JAK1) . | IC50 JAK2 (nM) (TYK2 > JAK2) . | IC50 JAK3 (nM) (TYK2 > JAK3) . |
|---|---|---|---|---|
| 33P-ATP | 0.6 ± 0.1 | 23 ± 19 (41) | 26 ± 12 (46) | 41 ± 11 (73) |
| TR-FRET | 13 ± 3 | 59 ± 43 (4.7) | 882 ± 502 (70.2) | 1437 ± 630 (114.3) |
| TR-FRET ruxolitinib | 6.5 | 1.1 | 31 | 196 |
| TR-FRET tofacitinib | 78 | 1.6 | 264 | 10 |
| pSTAT (cell lines) | 126 | 345 (2.7) | 1006 (8.4) | — |
| pSTAT (primary cells) | 107 | 423 (3.9) | 2223 (20.8) | 1608 (15.0) |
| . | IC50 TYK2 (nM) . | IC50 JAK1 (nM) (TYK2 > JAK1) . | IC50 JAK2 (nM) (TYK2 > JAK2) . | IC50 JAK3 (nM) (TYK2 > JAK3) . |
|---|---|---|---|---|
| 33P-ATP | 0.6 ± 0.1 | 23 ± 19 (41) | 26 ± 12 (46) | 41 ± 11 (73) |
| TR-FRET | 13 ± 3 | 59 ± 43 (4.7) | 882 ± 502 (70.2) | 1437 ± 630 (114.3) |
| TR-FRET ruxolitinib | 6.5 | 1.1 | 31 | 196 |
| TR-FRET tofacitinib | 78 | 1.6 | 264 | 10 |
| pSTAT (cell lines) | 126 | 345 (2.7) | 1006 (8.4) | — |
| pSTAT (primary cells) | 107 | 423 (3.9) | 2223 (20.8) | 1608 (15.0) |
Inhibition of JAK family members was assessed by ATP assay (33P-ATP) and TR-FRET–based STAT3 phosphorylation assay. For pSTAT-based cell assays, cells were incubated with SAR-20347 or DMSO for 20 min and then stimulated with cytokines indicated in Table II for 30 min (see 2Materials and Methods). Values represent the IC50 (nM). For the biochemical assays, the variation between three individual experiments is indicated. The fold selectivity for TYK2 is calculated by comparing the IC50 for TYK2 against each JAK family member.