TABLE 1

Bicifadine interactions with neurotransmitter transporters, receptors, and ion channels: radioligand-receptor binding screen The receptors tested, their sources, the radioligands used to define the binding sites, and the relative affinity of bicifadine for these receptors are indicated. Additional information on the assay conditions can be obtained from the contractor's website. Bicifadine showed significant affinity (Ki < 10 μM) for the α1-, α2-, and β1-adrenergic; 5-HT1A- and 5-HT1B-serotonergic; and σ1-receptors. Studies were performed at: 1Oregon Health Sciences University/VA Medical Center; 2Cerep; 3MDS Pharma Services; and 4Polish Academy of Sciences.


Transporter/Receptor/Ion Channel

Source

Assay Conditions (Radioligand, [nM]/nonspecific binding ligand, [μM])

Affinity
Norepinephrine transporter Human recombinant (HEK 293 cells) [125I]RTI-55, 0.04/mazindol, 51 Ki = 5.0 ± 1.1 μM
Serotonin transporter Human recombinant (HEK 293 cells) [125I]RTI-55, 0.04/imipramine, 51 Ki = 2.4 ± 0.7 μM
Dopamine transporter Human recombinant (HEK 293 cells) [125I]RTI-55, 0.04/mazindol, 51 Ki = 5.2 ± 1.2 μM
A1 Adenosine Human recombinant (CHO cells) [3H]DPCPX, 1/DPCPX, 12 <25% Inhibition at 10 μM
A2A Adenosine Human recombinant (HEK 293 cells) [3H]CGS 21680, 6/NECA, 102 <25% Inhibition at 10 μM
α1-Adrenergic Rat brain [3H]Prazosin, 0.25/prazosin, 0.13 Ki = 773 nM
α2-Adrenergic Rat cerebral cortex [3H]Rauwolscine, 0.7/yohimbine, 13 Ki = 1.01 μM
β1-Adrenergic Human recombinant (Rex 16 cells) [125I]Cyanopindolol, 0.03/S(-)-propranolol, 1003 Ki = 1.55 μM
β2-Adrenergic Human recombinant (CHO cells) [3H]CGP-12177, 0.2/ICI-118551, 103 41% Inhibition at 10 μM
β3-Adrenergic Human recombinant (HEK 293 cells) [125I]Cyanopindolol, 0.05/alprenolol, 10003 <25% Inhibition at 10 μM
Angiotensin1 Human recombinant (CHO cells) [125I][Sar1,Ile8]-Angiotensin II, 0.05/angiotensin II, 103 <25% Inhibition at 10 μM
Benzodiazepine Rat brain (- cerebellum) [3H]Flunitrazepam, 1/diazepam, 103 <25% Inhibition at 10 μM
Bradykinin1 Human IMR-90 cells [3H](des-Arg10)-Kallidin, 0.5/(Des-Arg9,Leu8)-bradykinin, 0.000173 <25% Inhibition at 10 μM
Bradykinin2 Human recombinant (CHO cells) [3H]Bradykinin, 0.2/bradykinin, 53 <25% Inhibition at 10 μM
Calcitonin gene-related peptide; Human SK-N-MC cells [125I]CGRP, 0.01/α-CGRP (8–37), 13 <25% Inhibition at 10 μM
Cannabinoid1 Human recombinant (HEK 293 cells) [3H]WIN 55212-2, 2/WIN 55212-2, 102 <25% inhibition at 10 μM
Cannabinoid2 Human recombinant (CHO-K1 cells) [3H]WIN 55212-2, 2.4/R(+)-WIN 55212-2, 0.00493 <25% Inhibition at 10 μM
CholecystokininA Human recombinant (NIH-3T3 cells) [3H]L-364718, 0.8/l-364718, 13 <25% Inhibition at 10 μM
Cholinergic receptor, M1 Muscarinic Human recombinant (CHO cells) [3H]N-Methylscopolamine 0.8/atropine, 13 <25% Inhibition at 10 μM
Cholinergic receptor, M2 Muscarinic Human recombinant (CHO cells) [3H]N-Methylscopolamine 0.8/atropine, 13 <25% Inhibition at 10 μM
Cholinergic receptor, M3 Muscarinic Human recombinant (CHO cells) [3H]N-Methylscopolamine 0.8/atropine, 13 38% Inhibition at 10 μM
Cholinergic receptor, M4 Muscarinic Human recombinant (CHO cells) [3H]N-Methylscopolamine 0.8/atropine, 13 <25% Inhibition at 10 μM
Cholinergic receptor, α7 Nicotinic Rat brain (- cerebellum) [125I]α-Bungarotoxin, 0.6/ α-bungarotoxin,13 <25% Inhibition at 10 μM
Corticotropin releasing factor1 Human recombinant (CHO-K1 cells) [125I](Tyr0)-CRF, 0.05/urocortin, 0.13 <25% Inhibition at 10 μM
D1 Dopamine Human recombinant (CHO cells) [3H]SCH 23390, 1.4/(+)-butaclamol, 103 <25% Inhibition at 10 μM
D2S Dopamine Human recombinant (CHO cells) [3H]Spiperone, 0,3/(+)butaclamol, 103 <25% Inhibition at 10 μM
D3 Dopamine Human recombinant (CHO cells) [3H]Spiperone, 0.7/S(-)-sulpiride, 253 <25% Inhibition at 10 μM
D4.4 Dopamine Human recombinant (CHO cells) [3H]Spiperone, 0.3/(+)butaclamol, 102 IC50 >30 μM
EndothelinA Human recombinant (CHO cells) [125I]Endothelin-1, 0.03/endothelin-1, 0.13 <25% Inhibition at 10 μM
Epidermal growth factor Human A431 cells [125I]EGF, 0.08/EGF, 0.13 <25% Inhibition at 10 μM
GABAA Rat brain (- cerebellum) [3H]Muscimol, 1/muscimol, 0.13 <25% Inhibition at 10 μM
Glucocorticoid Human HeLa S3 cells [3H]Dexamethasone, 6/dexamethasone, 203 <25% Inhibition at 10 μM
Glutamate receptor, AMPA type Rat cerebral cortex [3H]AMPA, 5/l-glutamic acid, 10003 <25% Inhibition at 10 μM
Glutamate receptor, NMDA type, agonist binding site Rat cerebral cortex [3H]CGP-39653, 2/l-glutamic acid, 10003 <25% Inhibition at 10 μM
Glutamate receptor, NMDA type, ionophore binding site Rat cerebral cortex [3H]MK-801, 5/PCP, 1004 Ki = 23.7 ± 7.9 μM
H1 Histamine Human recombinant (CHO cells) [3H]Pyrilamine, 1.2/pyrilamine, 13 44% Inhibition at 10 μM
Melanocortin4 Human recombinant (HEK-293 cells) [125I]NDP-α-MSH, 0.05/ NDP-α-MSH, 12 <25% Inhibition at 10 μM
Neurokinin1 U-373MG cells [3H]Sar9, Met(O2)11]-Substance P, 1.2/ Sar9, Met(O2)11]-SP, 12 <25% Inhibition at 10 μM
Neuropeptide Y1 Human SK-N-MC cells [125I]Peptide YY, 0.015/neuropeptide Y, 13 <25% Inhibition at 10 μM
μ Opioid Human recombinant (CHO cells) [3H]DAMGO, 0.5/naloxone, 102 IC50 >30 μM
κ Opioid Guinea-pig cerebellum [3H]U 69593, 0.7/naloxone, 102 IC50 >30 μM
δ2 Opioid Human recombinant (CHO cells) [3H]DADLE, 0.5/naltrexone, 102 <25% Inhibition at 30 μM
Orphanin1 Human recombinant (HEK 293 cells) [3H]Nociceptin, 0.6/orphanin-FQ, 13 <25% Inhibition at 10 μM
DP Prostanoid Human recombinant (CHO K1 cells) [3H]PGD2, 1.7/PGD2, 13 <25% Inhibition at 10 μM
EP2 Prostanoid Human recombinant (HKE-293 cells) [3H]PGE2, 4/PGE2, 103 <25% Inhibition at 10 μM
EP4 Prostanoid Human recombinant (HKE-293 cells) [3H]PGE2, 0.3/PGE2, 103 <25% Inhibition at 10 μM
P2X Purine NZW Rabbit urinary bladder [3H]α,β-Methylene-ATP, 8/β,γ-methylene-ATP, 1003 <25% Inhibition at 10 μM
5-HT1A Serotonin Human recombinant (HKE-293 cells) [3H]8-OH-DPAT, 0.5/8-OH-DPAT, 102 Ki = 1.7 μM
5-HT1B Serotonin Rat cerebral cortex [125I]Cyanopindolol, 0.1/serotonin, 102 Ki = 2.9 μM
5-HT3 Serotonin Human recombinant (HEK-293 cells) [3H]BRL 43694, 0.5/MDL 72222, 102 37% Inhibition at 10 μM
σ1 Jurkat T-cells [3H](+)Pentazocine, 8/haloperidol, 102 Ki = 1.2 μM
σ2 Rat cerebral cortex [3H]DTG, 5, (+)pentazocine, 300 nM/haloperidol, 102 Ki = 11 μM
Vanilloid1 Rat spinal cord [3H]Resiniferatoxin, 0.2/resiniferatoxin, 0.13 <25% Inhibition at 10 μM
Vasopressin1A Human recombinant (CHO cells) [3H]AVP, 0.3/AVP, 12 <25% Inhibition at 10 μM
α Ca2+ Channel Rat cerebral cortex [3H]Gabapentin, 20/gabapentin, 1003 <25% Inhibition at 10 μM
N-type Ca2+ Channel Rat frontal cortex [125I]ω-Conotoxin GVIA, 0.010/ω-conotoxin GVIA, 0.103 <25% Inhibition at 10 μM
KATP+ Channel Rat cerebral cortex [3H]Glibenclamide, 0.1/glibenclamide, 12 <25% Inhibition at 10 μM
Saxitoxin-sensitive KV+ channels Rat cerebral cortex [125I]α-Dendrotoxin, 0.01/α-dendrotoxin, 0.0502 <25% Inhibition at 10 μM
KV+ hERG
Human recombinant (HEK-293 cells)
[3H]Astemizole, 1.5/astemizole, 103
<25% Inhibition at 10 μM
  • DPCPX, 8-cyclopentyl-1,3-dipropylxanthine; CGS 21680, 4-[2-[[6-amino-9-(N-ethyl-β-d-ribofuranuronamidosyl)-9 H-purin-2-yl]amino]ethyl]benzenepropanoic acid HCl; NECA, 5′-N-ethylcarboxamidoadenosine; CGP-12177, 4-[3-[(1,1-dimethylethyl)amino]2-hydroxypropoxy]-1,3-dihydro-2H-be nzimidazol-2-one HCl; ICI-118551, (±)-1-[2,3-(dihydro-7-methyl-1H-inden-4-yl)oxy]-3-[(1-methylet hyl)amino]-2-butanol HCl; CGRP, calcitonin gene-related peptide; WIN 55212–2, (R)-(+)-[2,3-dihydro-5-methyl-3-(4-morpholinylmethyl)pyrrolo[1,2,3-de]-1,4-benzoxazin-6-yl]-1-napthalenylmethanone mesylate; L-364718, devazepide; CRF, corticotropin-releasing factor; SCH 23390, (R)-(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine HCl; EGF, endothelial growth factor; AMPA, α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid HBr; CGP-39653, d,l-(E)-2-amino-4-propyl-5-phosphono-3-pentenoic acid; PCP, 1-(1-phenylcyclohexyl)piperidine HCl; MSH, melanocyte-stimulating hormone; DAMGO, [d-Ala2,Nme-Phe4,Gly-ol5]-enkephalin; U 69593, (+)-(5α,7α,8β)-N-methyl-N-[7-(1-pyrrolidinyl)-1-ox aspiro[4.5]dec-8-yl]-benzeneacetamide; DADLE, [d-Ala2, d-Leu5]-enkephaline acetate; PGD2, prostaglandin D2; PGE2, prostaglandin E2; 8-OH-DPAT, (±)-8-hydroxy-2-dipropylaminotetralin HBr; BRL 43694, 1-methyl-N-(9-methyl-9-azabicyclo[3.3.1]non-3-yl)-indazole-3-carboxa mide; MDL 72222, tropanyl 3,5-dichlorobenzoate; DTG, 1,3-di-o-tolylguanine; AVP, arginine vasopressin