Effects of the mutations on [3H]osanetant displacement by RO4908594 in the membrane preparations from HEK293-EBNA cells transiently expressing WT and mutated hNK3Rs
Ki and nH values for [3H]osanetant binding inhibition by RO4908594 calculated as described under Materials and Methods. Values are mean ± S.E. of the Ki calculated from three independent experiments, each performed in duplicate. The mutations that affected the affinity constants of RO4908594 in comparison with WT are shown in bold. Statistical significance was determined using a two-tailed t test.
hNK3R | RO4908594 | ||||
|---|---|---|---|---|---|
| Ki | Ki (Mutant)/Ki (WT) | nH | |||
| nM | |||||
| WT | 1.6 ± 0.1 | 1.1 ± 0.1 | |||
| V95A | 1.6 ± 0.4 | 1.0 | 0.9 ± 0.1 | ||
| V95L | 3.5 ± 0.1 | 2.2 | 1.0 ± 0.0 | ||
| V95I/A99S | 4.2 ± 0.8 | 2.6 | 1.1 ± 0.1 | ||
| N138A | 4.0 ± 0.7 | 2.5 | 1.2 ± 0.0 | ||
| T139A | 2.9 ± 0.5 | 1.8 | 1.0 ± 0.1 | ||
| V95L/T139A | 4.9 ± 0.5 | 3.1 | 1.1 ± 0.0 | ||
| L232A | 1.6 ± 0.2 | 1.0 | 1.1 ± 0.0 | ||
| Y315F | 0.6 ± 0.04 | 0.4 * | 1.6 ± 0.1 | ||
| S341A | 2.0 ± 0.2 | 1.3 | 1.3 ± 0.3 | ||
| F342A | 0.02 ± 0.0 | 0.01 *** | 0.8 ± 0.2 | ||
| M346A | 6.0 ± 0.3 | 3.8 ** | 1.1 ± 0.1 | ||
| S348A |
8.0 ± 0.1
|
5.0
***
| 1.1 ± 0.1 | ||