Table 1 |.
DCLK1-IN-1 is an exquisitely selective inhibitor of DCLK1.
| Target | Assay (nM) | DCLK1-IN-1 (4) | DCLK1-NEG (9) | XMD8-92 (2) | LRRK2-IN-1 (1) | JQ1 | Trametinib |
|---|---|---|---|---|---|---|---|
| DCLK1 | ITC Kda | 55 | > 1000 | - | - | - | - |
| DCLK1 | binding assay IC50b | 9.5 | 2700 | 1400 | 3 | - | - |
| DCLK1 | kinase assay IC50c | 57.2 | > 10000 | 104 | 55 | - | - |
| DCLK1 | cellular assay IC50d | 279 | > 10000 | 1150 | 400 | - | - |
| DCLK2 | binding assay IC50e | 31 | 1400 | 3600 | 9.9 | - | - |
| DCLK2 | kinase assay IC50f | 103 | > 10000 | 385 | 45 | - | - |
| ERK5 | binding assay IC50g | 1800 | > 10000 | 1600 | 21 | - | - |
| ERK5 | kinase assay IC50h | 4000 | > 10000 | 364 | 160 | - | - |
| ERK5 | cellular assay IC50i | 5360 | > 10000 | 240 | 114 | - | - |
| LRRK2 | kinase assay IC50j | 6970 | > 10000 | 36 | 4.1 | - | - |
| BRD4 | binding assay IC50k | > 10000 | > 10000 | 1566 | 1347 | 37 | - |
| PATU-8988T cells | 2D monolayer culture IC50l | > 20000 | > 20000 | 2015 | 1151 | 67.4 | 60 (P.R.) |
| PATU-8988T cells | 3D spheroid culture IC50m | > 20000 | > 20000 | 6062 | 1866 | 111.2 | 1 |
| PATU-8902 cells | 2D monolayer culture IC50l | > 20000 | > 20000 | > 20000 | > 20000 | 1814 | 4.89 |
| PATU-8902 cells | 3D spheroid culture IC50m | > 20000 | > 20000 | > 20000 | > 1000 (P.R.) | > 1000 (P.R.) | < 0.63 |
| PANFR0172_T2 | 3D organoid culture IC50n | 877 | 8340 | > 20000 | 1340 | > 1000 (P.R.) | 791 |
| PANFR0172_T3 | 3D organoid culture IC50n | 2120 | > 20000 | > 20000 | 1500 | > 1000 (P.R.) | 1180 |
| PANFR0172_T4 | 3D organoid culture IC50n | > 20000 | > 20000 | > 20000 | > 20000 | > 1000 (P.R.) | > 20000 |
| PANFR0029_T2 | 3D organoid culture IC50n | > 20000 | > 20000 | 7030 | 1040 | 363 | 185 |
ITC data against purified recombinant DCLK1 protein. Data are representative of n = 2 independent experiments.
DCLK1 KINOMEscan binding assay. Data are presented as mean of n = 2 biologically independent samples
33P-ATP DCLK1 kinase assay. Assays were performed at an ATP concentration of 50 μM (Km). Data are presented as mean of n = 3 biologically independent samples.
DCLK1 nanoBRET assay. Data are presented as mean of n = 4 biologically independent samples and are representative of n = 3 independent experiments.
DCLK2 KINOMEscan binding assay. Data are presented as mean of n = 2 biologically independent samples (f) 33P-ATP DCLK2 kinase assay. Assays were performed at an ATP concentration of 100 μM (Km). Data are presented as mean of n = 3 biologically independent samples.
ERK5 KINOMEscan binding assay. Data are presented as mean of n = 2 biologically independent samples and
ERK5 32P-ATP kinase assay. Data are presented as mean of n = 3 biologically independent samples and representative of n = 2 independent experiments.
ERK5 cellular kinase assay. Data are presented as mean of n = 2 biologically independent samples and are representative of n = 2 independent experiments.
LRRK2 ADAPTA kinase assay. Data are presented as mean of n = 2 biologically independent samples.
BRD4 AlphaScreen assay. Data are presented as the mean of n = 3 biologically independent samples and representative of n = 2 independent experiments.
Analysis of cell viability in 2D-adherent cultures (l) and ultra-low adherent 3D-spheroid suspensions (m). Data in l and m are presented as mean of n = 4 biologically independent samples and are representative of n = 3 independent experiments.
Analysis of patient-derived pancreatic organoid viability. Data are presented as mean of n = 3 biologically independent samples and are representative of n = 2 independent experiments.
P.R., partial response.