Table 2.
Targets and off-targets of AZD1208 and HS38. Possible (off-)targets involved in anticontractile effects observed with AZD1208 and HS38 in this study, suggested by combination and alignment of previous data for affinities and IC50 values, with applied concentrations and findings in our experiments. See main text for references reporting available affinities and inhibition data from pharmacological profiling of both compounds
| AZD1208 | HS38 | |||||
|---|---|---|---|---|---|---|
| Kd 1 | IC50 2 | Kd | IC50 3 | Inhibition (10 µM) 3 | Ki | |
| Pim-1 | 0.2 nM | 2.6 nM | 1.8 µM | 1.7 µM | 77% 3, ca. 60% 4 | 2.94 µM |
| Pim-2 | 0.88 nM | 150–164 nM | 6.5 µM | 18 µM | 36% 3, ca. 10% 4 | > 100 µM |
| Pim-3 | 0.76 nM | 9–17 nM | 810 nM | 200 nM | 76% 3, ca. 85% 4 | 208 nM |
| DAPK1 | 420 nM | 300 nM | 200 nM | 89% 3 | ||
| DAPK2 | Probably > > 10 µM | 79 nM | ||||
| ZIPK/DAPK3 | Probably > > 10 µM | 280 nM | ca. 85% 4 | 260 nM | ||
| MLCK | Probably > > 10 µM | 19 µM | 29% 3 | |||
| ROCK | Probably > > 10 µM | 200 µM | 9% 3 | |||
| Off-targets | CDK7 38 nM, MAPK15 53 nM, CAMK4 360 nM, HIPK3 480 nM, STK17B 930 nM, seven further kinases > 10 µM | IRAK4 1.4 µM, DYRK2 1.8 µM | IRAK4 80%, DYRK2 79% 3 | |||
| Prostate tissues | α1-adrenergic contractions inhibited by 500 nM, but not by 100 nM | α1-adrenergic contractions only slightly and partly inhibited by 3 µM | ||||
| Interlobar arteries | α1-adrenergic contractions partly inhibited by 500 nM | α1-adrenergic contractions partly inhibited by 3 µM | ||||
| Putative (off-)targets involved in anticontractile effects 5 | DAPK1, CAMK4, HIPK3 (Kd/IC50 > > 100 nM, < 900 nM). Pim kinases: no, because no inhibition with 100 nM | ZIPK, IRAK4, DYRK2 (inhibition with 3 µM expected). Pim-1, -3: no, because no inhibition by 100 nM AZD1208. DAPK1: probably no, because no inhibition in prostate, but possible target for AZD1208 in prostate, while targets of AZD1208 and HS38 must be unshared between prostate and interlobar artery | ||||
1Determined in follow-up, after 13 off-target kinases were identified (inhibited > 50% by unknown concentration of AZD1208) by screening of 442 kinases
2In biochemical assays with 5-mM ATP (= high end of physiologic ATP concentrations in human cells)
3Biochemical assays using 124 different, purified kinases, not including ZIPK
4Biochemical assay, analog to (3)
5By fusion and intersection of organ bath data in this study with data from pharmacological profiling