Table 4.

Effects of the investigated analogues in inhibiting the dissociation of [³H]S-1 from hSERT WT. The relative potency of 30 µM compound in inhibiting [³H]S-1 dissociation as compared to buffer and S-1.

Compound	[3H] S-1 dissoc. t½ (min) at 18°C	[3H] S-1 dissoc. t½ (min) at 24°C	24°C dissociation t½ calculated to a t½ at 18°C (min)
Control	16.1±1.0	7.0±0.3
S-1	68±12	28.1±3.1
5	17.4±0.8
8	16.4±0.3
9	16.3±0.1
10	24.8±1.1
12	22.0±1.2
13	22.1±1.5
14	11.4±0.2
15	21.4±0.3
19		40.2±7.0	97±17
20		52.6±4.1	127±10
21	23.1±2.9
22	33.2±5.8
24	24.7±5.4
25		63±13	151±30
26	44±12
42		45.4±5.9	110±14
45	59.6±1.0
46	41.2±2.8
48	29.0±1.3
49	60.1±1.0
51		52.3±3.2	127.1±6.8
53		93±15	225±37
55		58.7±10.0	143±24
60	52.0±7.6
61	43.1±6.2
(+)62		54.9±3.5	132.3±8.5
(−)62	37.3±9.5
63	34.4±6.6

Experiments were performed on membrane preparations from COS-7 cells transiently expressing with hSERT WT. [³H]S-1 was added until equilibrium was obtained and subsequently diluted in 12× buffer volumes containing 30 µM of the indicated compound. Dissociation were measured at 18°C to obtain a suitable t½ (<100 min). For the compounds with t½ >100 min, the temperature were raised to 24°C for a more accurate determination. Since the dissociation t½ is directly proportional to the temperature, it is possible to calculate a t½ at 18°C from the 24°C dissociation. This temperature constant is found for the effect of S-1 on [³H]S-1 dissociation at 18°C and 24°C, respectively. Values are mean±S.E. of 3–6 experiments performed in triplicate.