Table 1.
Kinetic constants of substrates and inhibitors of B0AT1
| FLIPR assay | Radioactive uptake | |||||
|---|---|---|---|---|---|---|
| Compound | NSC | Purity (%) | KM | IC50 | KM | IC50 |
| Leucine | >98 | 500 ± 100 | 1050 ± 50 | |||
| Isoleucine | >98 | 700 ± 60 | 1400 ± 100 | |||
| S‐Benzyl‐L‐cysteine | 97 | 14 ± 2 | 398 ± 29 | |||
| O‐Benzyl‐L‐serine | >99 | 14 ± 5 | 810 ± 176 | |||
| S‐(4‐Tolyl)‐L‐cysteine | >95 | 23 ± 14 | 205 ± 46 | |||
| S‐Phenyl‐L‐cysteine | >97 | 49 ± 23 | ||||
| Nimesulide | >98 | 172 ± 26* | ||||
| Benztropine | 63 912 | >98 | 44 ± 9 | 71 ± 8 | ||
| 2‐Benzyl‐1‐(3‐phenylpropyl)piperidine | 22 789 | >98 | 90 ± 21 | 78 ± 16 | ||
| 1‐(1,2‐diphenylethyl)‐3‐methylpiperidine | 34 358 | >95 | 5 ± 1 | 17 ± 3* | ||
| 11‐dihydro‐5H‐dibenzo[a,d]cyclohepten‐5‐ylidene)methyl]‐1‐methyl‐pyrrolidine | 169 092 | >99 | 10 ± 9 | |||
| 1‐(4,4‐diphenylbut‐3‐enyl)piperidine | 39 706 | >99 | 13 ± 3 | 70 ± 21 | ||
| 4‐(dibenzo[1,2‐a:1′,2′‐e][7] annulen‐11‐ylidene)‐1‐methylpiperidine | 169 911 | >99 | 15 ± 3 | 67 ± 16* | ||
| 3‐[(2‐aminophenyl)sulfanyl]‐ 1‐[4‐(4‐chlorophenoxy) phenyl]pyrrolidine‐2,5‐dione | 201 503 | >99 | 17 ± 10 | |||
| N1,n1‐dimethyl‐4‐[[4‐(dimethylamino)phenyl](4‐nitrophenyl)methyl]aniline | 3323 | 94 | 33 ± 13 | |||
| 2‐chloro‐N‐(1,1,3,3,5‐ pentamethyl‐6‐(1‐pyrrolidinyl) ‐2,3‐dihydro‐1H‐inden‐4‐yl)benzamide | 321 496 | 92 | 37 ± 23 | 55 ± 22* | ||
Concentrations are given in μM. All compounds were preincubated with the cells for >30 min in the FLIPR assay. Some inhibitors required preincubation for 4 h to be effective (indicated by *). In the FLIPR assay, 1.5 mM leucine was used as substrate (n = 3); in the radioactive uptake assay, 150 μM isoleucine was used as substrate (n = 3). The chemical identification number (NSC) is shown for compound identification. Compound purity as provided by supplier or determined by liquid chromatography (see Supporting Information Data S1 and S2)