Table 2.
Competition binding affinities of different ligands obtained with [3H]RX821002 at wild-type and chimaeric α2-adrenoceptors expressed in CHO cells
| Human α2A | ||||||||
|---|---|---|---|---|---|---|---|---|
| Ligand | WT (AAA) | nH | BBA | nH | CCA | nH | ACA | nH |
| ARC239 | 1600 (1000–2600) | 0.89 | 450 (160–1400) | 0.66 | 820 (600–1110) | 0.72 | 760 (480–1200) | 1.02 |
| Prazosin | 1500 (800–3100) | 0.84 | 1400 (490–2400) | 1.16 | 820 (600–1100) | 0.94 | 910 (600–1500) | 0.89 |
| Spiperone | 1200 (870–1600) | 0.94 | 870 (450–1800) | 0.93 | 560 (440–730)a | 1.03 | 500 (320–770)aa | 0.86 |
| Spiroxatrine | 550 (240–1300) | 1.33 | 69 (34–140)aa | 0.92 | 71 (39–130)aa | 1.08 | 91 (48–180)aa | 1.51 |
| Chlorpromazine | 600 (430–870) | 0.82 | 160 (83–310)aaa | 0.89 | 120 (56–260)aaa | 0.68 | 160 (100–230)aaa | 1.07 |
| Clozapine | 89 (54–150) | 0.91 | 67 (32–150) | 0.69 | 34 (17–67) | 0.58 | 51 (35–77) | 1.13 |
| Atipamezole | 2.1 (1.5–3.1) | 0.89 | 2.0 (0.99–4.3) | 1.00 | 1.2 (0.95–1.5) | 0.82 | 1.1 (0.77–1.5) | 0.97 |
| Idazoxan | 22 (15–31) | 0.80 | 15 (2.7–45) | 0.97 | 17 (13–24) | 0.91 | 13 (8.3–22) | 0.85 |
| Rauwolscine | 1.8 (0.78–4.5) | 0.53 | 4.4 (2.0–9.8) | 0.66 | 0.85 (0.40–2.1) | 1.02 | 1.6 (0.82–3.1) | 0.71 |
| Human α2B | ||||||
|---|---|---|---|---|---|---|
| Ligand | WT (BBB) | nH | AAB | nH | CCB | nH |
| ARC239 | 150 (40–570)bbb | 0.74 | 240 (120–480) | 0.72 | 98 (48–220) | 0.74 |
| Prazosin | 47 (17–140)bb | 0.79 | 200 (120–360) | 0.82 | 170 (89–310) | 0.89 |
| Spiperone | 12 (3.7–38)bbb | 0.51 | 26 (11–65) | 1.65 | 14 (6.0–38) | 0.67 |
| Spiroxatrine | 2.4 (1.2–5.0)bbb/cc | 0.64 | 6.8 (1.8–28) | 2.03 | 1.6 (0.64–5.9) | 0.91 |
| Chlorpromazine | 43 (20–100)bbb/cc | 0.64 | 53 (20–200) | 1.84 | 46 (18–120) | 0.70 |
| Clozapine | 12 (5.0–28)bbb | 0.93 | 18 (8.7–45) | 1.09 | 6.7 (1.9–23) | 0.64 |
| Atipamezole | 2.7 (0.56–14) | 0.85 | 3.0 (1.7–5.6) | 0.86 | 1.7 (0.80–4.0) | 0.65 |
| Idazoxan | 24 (18–34) | 1.08 | 37 (29–48) | 0.95 | 30 (19–51) | 0.88 |
| Rauwolscine | 1.1 (0.7–1.8) | 0.97 | 1.2 (0.66–2.4) | 1.25 | 2.7 (1.1–6.9) | 1.02 |
| Human α2C | |||||||
|---|---|---|---|---|---|---|---|
| Ligand | WT (CCC) | nH | AAC | nH | BBC | CAC | nH |
| ARC239 | 130 (66–260)ddd | 1.15 | 180 (86–380) | 0.89 | n.d. | 77 (34–180) | 0.65 |
| Prazosin | 45 (21–81)dd | 0.67 | 110 (50–230) | 0.78 | n.d. | 120 (55–260) | 0.56 |
| Spiperone | 29 (9.2–81)ddd | 0.66 | 26 (10–75) | 0.63 | n.d. | 39 (18–89) | 0.79 |
| Spiroxatrine | 13 (5.9–28)ddd | 1.13 | 12 (5.8–27) | 1.05 | n.d. | 18 (8.4–31) | 0.68 |
| Chlorpromazine | 260 (150–350)dd | 0.81 | 260 (110–720) | 0.42 | n.d. | 230 (130–400) | 0.80 |
| Clozapine | 6.5 (2.5–18)ddd | 0.75 | 5.1 (2.6–8.2) | 0.57 | n.d. | 1.8 (0.2–17) | 0.33 |
| Atipamezole | 4.1 (2.0–9.0) | 0.92 | 5.3 (4.8–8.3) | 0.82 | n.d. | 10 (5.0–23) | 1.16 |
| Idazoxan | 71 (20–251) | 0.71 | 48 (21–110) | 0.73 | n.d. | 18 (12–34) | 0.89 |
| Rauwolscine | 0.47 (0.21–1.2) | 0.83 | 0.27 (0.12–0.63) | 0.78 | n.d. | 0.39 (0.20–0.83) | 1.14 |
The apparent Ki (nM) and their 95% confidence intervals from three to six independent experiments, analysed using a one-site competition model. The pseudo-Hill slope (nH) was obtained using a variable slope model. Receptors are named based on the exchanged domains (see Figure 1). Statistical significances are shown with symbols (one) P < 0.05; (two) P < 0.01; (three) P < 0.001. (a) chimaera versus wild-type α2A; (b) wild-type α2A versus wild-type α2B; (c) wild-type α2B versus wild-type α2C; (d) wild-type α2A versus wild-type α2C.