TABLE 1

Binding affinities of CM156

Affinities (K_i values in nanomolar) were determined in tissue or cell homogenates. The values represent the mean ± S.E.M. from replicate assays. Values of >10,000 signify that there was less than 30% displacement of the radioligand at that concentration.

	Radioligand	Nonspecific Binding	Tissue	K_i
σ Receptors
σ₁	5 nM [³H](+)-Pentazocine	10 μM Haloperidol	Rat brain	1.3 ± 0.4
σ₂	3 nM [³H]Di-o-tolylguanidine	10 μM Haloperidol	Rat brain	0.6 ± 0.1
Monoamine transporters
Dopamine	0.5 nM [³H]WIN 35,428	50 μM Cocaine	Rat striatum	1175 ± 100
Serotonin	0.2 nM [³H]Paroxetine	1.5 μM Imipramine	Rat brainstem	1402 ± 152
Norepinephrine	0.5 nM [³H]Nisoxetine	4 μM Desipramine	Rat cerebral cortex	>10,000
Other neurotransmitter receptors
Adenosine	4.0 nM [³H]5′-N-Ethylcarboxamidoadenosine	1 μM 5′-N-Ethylcarboxamidoadenosine	Bovine striatum	>10,000^{^a}
Adrenergic, α₁	0.3 nM [³H]7-MeOxy-prazosin	1 μM Phentolamine	Rat forebrain	695^{^a}
Adrenergic, α₂	1 nM [³H]RX 821002	1 μM Phentolamine	Rat cerebral cortex	>10,000^{^a}
Adrenergic, β₁	0.04 nM [¹²⁵I](−)-Iodocyanopindolol	3 μM Alprenolol	Human neuroepithelioma	>10,000^{^a}
Cannabinoid, CB₁	0.5 nM [³H]CP 55940	1 μM HU-210	Human recombinant embryonic kidney 293 cells	>10,000^{^a}
Cannabinoid, CB₂	0.5 nM [³H]CP 55940	1 μM HU-210	Human recombinant CHO cells	>10,000^{^a}
Dopamine, nonselective	0.3 nM [³H]Spiperone	1 μM Spiperone	Bovine striatum	>10,000^{^a}
Dopamine D₁	0.18 nM [³H]SCH 23390	1 μM SCH 23390	LHD₁ cells	>10,000^{^a}
Dopamine D₂	5 nM [³H](−)-Sulpiride	1 μM Haloperidol	Rat brain	1041 ± 9
	0.21 nM [³H]YM-09151-2	1 μM Chlorpromazine	CHOp cells	>10,000^{^a}
Dopamine D₃	0.21 nM [³H]YM-09151-2	1 μM Chlorpromazine	CHOp cells	>10,000^{^a}
Dopamine D_4.2	0.15 nM [³H]Spiperone	1 μM Haloperidol	Human recombinant CHO-K1 cells	53^{^a}
GABA_A, agonist site	5 nM [³H]GABA	1 μM GABA	Bovine cerebellum	>10,000^{^a}
GABA_A, benzodiazepine α₁	1 nM [³H]Flunitrazepam	0.5 μM Flumazenil	Bovine cortex	>10,000^{^a}
GABA_B	1 nM [³H]CGP 54626A	100 μM Baclofen	Rat cerebral cortex	>10,000^{^a}
Glutamate, AMPA	5 nM [³H]AMPA	100 μM AMPA	Rat forebrain	>10,000^{^a}
Glutamate, kainate	10 nM [³H]Kainic acid	10 μM Kainic acid	Rat forebrain	>10,000^{^a}
Glutamate, NMDA agonist	2 nM [³H]CGP 39653	300 μM NMDA	Rat forebrain	>10,000^{^a}
Glutamate, NMDA glycine	4 nM [³H]MDL-105,519	3 μM MDL-105,519	Rat cortex/hippoc	>10,000^{^a}
Glutamate, NMDA/phencyclidine	10 nM [³H]TCP	100 μM (+)-MK801	Rat forebrain	>10,000^{^a}
	5 nM [³H]TCP	10 μM Cyclazocine	Rat brain	>10,000
Glutamate, mGluR1	20 nM [³H]Quisqualic acid	1 mM l-Glutamate	Rat cerebellum	>10,000^{^a}
Glutamate, mGluR5	10 nM [³H]2-Methyl-6-(phenylethynyl)pyridine	10 μM 2-Methyl-6-(phenylethynyl)pyridine	Rat brain	>10,000^{^a}
Glycine, strychnine	16 nM [³H]Strychnine	100 μM Strychnine nitrate	Rat spinal cord	>10,000^{^a}
Histamine H₁	2 nM [³H]Pyrilamine	10 μM Triprolidine	Bovine cerebellum	853^{^a}
Histamine H₂	0.1 nM [¹²⁵I]aminopotentidine	3 μM Tiotidine	Guinea pig striatum	2570^{^a}
Histamine H₃	0.2 nM [³H]N-α-Methylhistamine	0.1 nM R-(−)-α-Methylhistamine	Rat forebrain	2385^{^a}
Melatonin	70 pM [¹²⁵I]2-Iodomelatonin	0.1 μM 2-Iodomelatonin	Chicken brain	>10,000^{^a}
Muscarinic, central	0.15 nM [³H]Quinuclidinyl benzilate	0.1 μM Atropine	Rat cerebral cortex	3080^{^a}
Muscarinic, peripheral	0.3 nM [³H]Quinuclidinyl benzilate	0.1 μM Atropine	Guinea pig bladder	2155^{^a}
Muscarinic, M₁	0.5 nM [³H]N-Methyl scopolamine	1 μM (−)-Scopolamine	Human recombinant CHO cells	>10,000^{^a}
Muscarinic, M₂	0.5 nM [³H]N-Methyl scopolamine	1 μM Methylscopolamine	Human recombinant CHO cells	1210^{^a}
Nicotinic, muscle	1 nM ¹²⁵I-α-Bungarotoxin	10 μM Nicotine	Human TE671 cells	>10,000^{^a}
Nicotinic, neuronal	0.05 nM [³H]Epibatidine	20 nM Epibatidine	Human SK-N-F1 cells	>10,000^{^a}
Opioid, nonselective	0.5 nM [³H]Bremazocine	10 μM Levallorphan	Rat brain	>10,000
	1 nM [³H]Naloxone	1 μM Naloxone	Rat forebrain	>10,000^{^a}
Serotonin, nonselective	5 nM [³H]Lysergic acid diethylamide	10 μM Methysergide	Rat cerebral cortex	2060^{^a}
5-HT_1A	0.50 nM [³H]8-Hydroxy-2-dipropylaminotetralin	1 μM Dihydroergotamine	HA₇ cells	4985 ± 330^{^a}
5-HT₂	2 nM [³H]Ketanserin	1 μM Mianserin	Rat brain	1376 ± 159
5-HT_2A	0.40 nM [³H]Ketanserin	1 μM Ketanserin	NIH-3T3-GF₆ cells	1129 ± 49^{^a}
5-HT_2C	0.40 nM [³H]Mesulergine	10 μM Mesulergine	NIH-3T3-Po cells	>10,000^{^a}
Hormones, peptides, steroids
Angiotensin II, AT₁	0.06 nM [¹²⁵I](Sar¹-Ile⁸)angiotensin	1 μM Angiotensin II	Human KAN-TS cells	>10,000^{^a}
Angiotensin II, AT₂	0.1 nM [¹²⁵I]Tyr⁴-angiotensin II	0.05 μM Angiotensin II	Bovine cerebellum	>10,000^{^a}
Bradykinin, BK₂	0.2 nM [³H]Bradykinin	100 nM Bradykinin trifluoroacetic acid	Guinea pig ileum	>10,000^{^a}
Cholecystokinin, CCK_A	0.02 nM [¹²⁵I]CCK-8	1 μM CCK-8	Mouse pancreas	>10,000^{^a}
Cholecystokinin, CCK_B	0.02 nM [¹²⁵I]CCK-8	1 μM CCK-8	Mouse forebrain	>10,000^{^a}
CRF, nonselective	0.1 nM [¹²⁵I]Tyr⁰-oCRF	1 μM Tyr⁰-oCRF	Rat cerebral cortex	>10,000^{^a}
Endothelin, ET_A	0.033 nM ¹²⁵I-Endothelin-1	0.1 μM Endothelin-1	Human neuroblastoma	>10,000^{^a}
Endothelin, ET_B	0.025 nM ¹²⁵I-Endothelin-1	0.1 μM Endothelin-1	Human astrocytoma	>10,000^{^a}
Estrogen	0.1 nM ¹²⁵I-3,7β-Estradiol	10 nM 17β-Estadiol	Human breast cancer	>10,000^{^a}
Galanin, nonselective	0.07 nM ¹²⁵I-Galanin	100 nM Galanin (porcine)	Rat brain	>10,000^{^a}
Glucocorticoid	1 nM [³H]Dexamethasone	10 μM Triamcinolone	Human recombinant	>10,000^{^a}
Neurokinin, NK₁	1.4 nM [³H]Substance P	1 μM Substance P	Rat submaxillary gland	>10,000^{^a}
Neurokinin, NK₂	0.1 nM [¹²⁵I-Neurokinin A	1 μM Neurokinin A	Human recombinant CHO cells	1510^{^a}
Neurokinin, NK₃ (NKB)	0.1 nM ¹²⁵I-Eledoisin	1 μM Eledoisin	Rat cerebral cortex	>10,000^{^a}
Oxytocin	1 nM [³H]Oxytocin	1 μM Oxytocin	Rat uterus	>10,000^{^a}
Testosterone, cytosolic	0.5 nM [³H]Methyltrienolone	0.7 μM Methyltrienolone	Human LnCAP cells	>10,000^{^a}
TRH	2 nM [³H](3MeHis²)TRH	10 μM TRH	Rat forebrain	>10,000^{^a}
VIP, nonselective	0.05 nM [¹²⁵I]VIP	1 μM VIP	Rat forebrain	>10,000^{^a}
Vasopressin 1	0.5 nM [³H]Phenylalanyl-3,4,5-v	1 μM Arg⁸-Vasopressin	Rat liver	>10,000^{^a}
Ion channels:
Calcium, type L (benzothiazepine)	5 nM [³H]cis-(+)-Diltiazem	10 μM Diltiazem	Rat cerebral cortex	751^{^a}
Calcium, type L (dihydropyridine)	0.2 nM [³H]Nitrendipine	1 μM Nifedipine	Rat cerebral cortex	1710^{^a}
Calcium, type N	0.01 nM ¹²⁵I-ω-Conotoxin GVIA	0.1 μM ω-Conotoxin GVIA	Rat cerebral cortex	>10,000^{^a}
Chloride, GABA (t-butylbicycloorthobenzoate)	20 nM [³H]t-Butylbicycloorthobenzoate	10 μM t-Butylbicyclophosphorothionate	Rat cerebral cortex	>10,000^{^a}
Potassium, ATP-sensitive	0.2 nM [³H]Glibenclamide	0.1 μM Glibenclamide	Rat cerebral cortex	>10,000^{^a}
Potassium, Ca²⁺ act VI	0.05 nM ¹²⁵I-Apamin	100 nM Apamin	Rat forebrain	>10,000^{^a}
Potassium, human ether-a-go-go-related gene	40 nM [³H]Astemizole	100 μM Terfenadine	CHO cells	>10,000^{^a}
Sodium, site 2	2 nM [³H]Batrachotoxin	1 mM Aconitine	Rat forebrain	1535^{^a}
Enzymes and other miscellaneous
Acetylcholine esterase	0.3 mM [³H]Acethylthiocholine	100 μM Physostigmine	Human recombinant	>10,000^{^a}
Choline acetyltransferase	0.2 nM [¹⁴C]Acetyl coenzyme	0.1 μM Ro 41-1049	Rat cerebral cortex	>10,000^{^a}
Glutamic acid decarboxylase	4 μM [¹⁴C]l-Glutamic acid	100 μM Aminooxy acetic acid	Rat striatum	>10,000^{^a}
Leukotriene, LTB₄ (BLT)	0.48 nM [³H]Leukotriene B₄	500 nM Leukotriene B₄	Guinea pig spleen	>10,000^{^a}
Leukotriene, LTD₄ (CysLT1)	0.2 nM [³H]Leukotriene D₄	1 μM Leukotriene D₄	Guinea pig lung	>10,000^{^a}
MAO_A oxidase, peripheral	50 μM [¹⁴C]5-HT	1 μM Ro 41-1049	Rat liver mitochondria	>10,000^{^a}
MAO_B oxidase, peripheral	10 μM [¹⁴C]Phenylethylamine	10 μM Ro 16-6491	Rat liver mitochondria	>10,000^{^a}
Nitric-oxide synthase (NOS)	5 nM [³H]l-N^G-Nitro-arginine	100 μM l-N^G-Nitro-arginine	Rat brain	1220^{^a}
Platelet activating factor	1.7 nM [³H]Hexadecyl-acetyl-platelet-activating factor	1 μM C₁₆-platelet-activating factor	Rabbit platelets	>10,000^{^a}
Thromboxane, TXA₂	2 nM [³H]SQ 29,548	10 μM Pinane-thromboxane	Human platelets	>10,000^{^a}

CHO, Chinese hamster ovary; CRF, corticotrophin-releasing factor; TRH, thyrotropin releasing hormone; VIP, vasoactive intestinal peptide.
↵a These values were determined by NovaScreen. All other values were determined in-house using the assay conditions described under Materials and Methods.