Supplementary Material Table 1 - Customized in Vitro Screening of L-Stepholidine by Novascreen

Supplementary Material Table 1 - Customized in vitro screening of l-stepholidine by Novascreen®

Receptor / Species / Tissue / Radioligand
(Concentration in nM
used in the assay) / Kd(M) / Reference compound / Ki (M)
(of the reference compound) / % inhibition of specific activity by
l-stepholidine (10E-7M)* / Novascreen
Catalog No.
Neurotransmitters
GABA A,
Agonist site / Bovine / Cerebellum / [3H]GABA (5.0) / 3.7E-7 / GABA / 1.24E-8 / 13.87% / 100-0021
GABA A,
BDZ, alpha 1 site / Bovine / Cortex / [3H]Flunitrazepam (1.0) / 1.4 E-9 / Ro15-1788 / 1.52E-9 / -13.48% / 100-0022
GABA-B / Rat / Cortex / [3H]CGP 54626A (1.0) / 1.2 E-9 / (+/-)Baclofen / 1.38E-6 / 10.75% / 100-0024
Glutamate,
AMPA site / Rat / Forebrain / [3H]AMPA (5.0) / 2.8E-8 / (+/-)AMPA Hbr / 2.98E-8 / -6.02% / 100-0026
Glutamate,
Kainate site / Rat / Forebrain / [3H]Kainic acid (10.0) / 16.0 E-9 / Kainic acid / 6.6E-9 / -18.84% / 100-0027
Glutamate,
NMDA Agonist site / Rat / Forebrain / [3H]CGP 39653 (2.0) / 7.0 E-9 / NMDA / 9.11E-6 / 0.95% / 100-0029
Glutamate,
NMDA Glycine (Stry-insensitive site) / Rat / Cortex / [3H]-MDL-105,519 (4.0) / 2.0E-8 / MDL-105,519 / 1.57E-8 / 6.19% / 100-0032
Glycine,
Strychnine-sensitive / Rat / Spinal Cord / [3H]Strychnine (16.0) / 28.0 E-9 / Strychnine nitrate / 1.74E-7 / -9.52% / 100-0033
Muscarinic, M1 (hr) / Human / CHO cell line / [3H]Scopolamine,N-Methyl (0.5) / 5.0E-11 / (-)Scopolamine MeBr / 9.09E-11 / 2.72% / 100-0039
Muscarinic, M2 (hr) / Human / CHO cell line / [3H]Scopolamine,N-Methyl (0.5) / 2.9E-10 / (-)Scopolamine MeBr / 7.59E-10 / 8.33% / 100-0039
Muscarinic,
Non-selective, Central / Rat / Cortex / [3H]QNB (0.15) / 1.0E-10 / Atropine sulphate / 2.78E-10 / -3.02 / 100-0135
Muscarinic,
Non-selective, Peripheral / G.pig / Bladder / [3H]QNB (0.3) / 0.3E-9 / Atropine sulphate / 2.86E-10 / 9.46% / 100-0136
Nicotinic (-Bungarotoxin insensitive) / Rat / Cortex / [3H]Epibatidine (0.1) / 63.0E-12 / (+/-)Epibatidine / 1.39E-10 / 10.31% / 100-0046

* Values represent the average of replicate tubes tested with 10E-7M of l-stepholidine; bolded values represent inhibition beyond baseline range (-20% to +20%) inhibition of binding or enzymatic activity. More details on individual assays can be obtained from Novascren® Biosciences Ltd. USA., website

Receptor / Species / Tissue / Radioligand
(Concentration in nM
used in the assay) / Kd(M) / Reference compound / Ki (M)
(of the reference compound) / % inhibition of specific activity by
l-stepholidine (10E-7M)* / Novascreen
Catalog No.
Neurotransmitter transporters
Dopamine Transporter / G.pig / Striatum / [3H]WIN 35,428 (6.0) / 28.0E-9 / GBR12909 / 8.27E-9 / -0.81% / 100-0015
Norepinephrine transporter / Rat / Forebrain / [3H]Nisoxetine (1.0) / 0.9E-9 / Desimipramine HCl / 1.22E-9 / 11.53% / 100-0048
Serotonin transporter / Rat / Cortex / [3H]Citalopram, N-Methyl (0.7) / 1.7 E-9 / Imipramine HCl / 2.93E-8 / -1.79% / 100-0056
Ion channels
Calcium channel, Type L (Dihydropyridine site) / Rat / Cortex / [3H]Nitrendipine (0.2) / 0.2E--9 / Nifedipine / 8.76E-10 / 11.07% / 100-0013
Calcium channel, Type N / Rat / Cortex / [125I]Conotoxin GVIA (0.01) / 1.0E-11 / -Conotoxin GVIA / 1.05E-11 / -3.01% / 100-0014
Potassium channel, ATP-sensitive / Rat / Cortex / [3H]Glibenclamide (0.2) / 0.25E-9 / Glibenclamide / 4.8E-10 / 0.44% / 100-0052
Potassium channel, Ca2+ Act. VI / Rat / Cortex / [125I]Apamin (0.05) / 7.0E-11 / Apamin / 3.61E-11 / 8.25% / 100-0054
Potassium channel, I(Kr) (hERG) (hr) / Human / CHO cell line / [3H]Astemizole (40) / 100E-9 / Terfenadine / 7.34E-7 / 10.52% / 100-0330
Sodium site 2 / Rat / Forebrain / [3H]Batrachotoxin A
20--benzo (2.0) / 32.0E-9 / Aconitine / 1.77E-6 / 6.84% / 100-0069

Receptor / Species / Tissue / Radioligand
(Concentration in nM
used in the assay) / Kd(M) / Reference compound / Ki (M)
(of the reference compound) / % inhibition of specific activity by
l-stepholidine (10E-7M)* / Novascreen
Catalog No.
Brain/Gut Peptides
Angiotensin II,
AT1 (h) / Human / KAN – TS cell line / [125I]-(Sar1-Ile8) Angiotensin (0.06) / 8.5E-10 / Angiotensin II (human) / 5.13E-9 / 1.58% / 100-0084
Angiotensin II (central), AT2 / Bovine / Cerebellum / [125I]Tyr4-Angiotensin II / 0.4E-9 / Angiotensin II (human) / 3.67E-10 / -13.21% / 100-0108
Bradykinin, BK2 / G.pig / Ileum / [3H]Bradykinin (0.2) / 0.4E-9 / Bradykinin TFA / 2.43E-10 / 5.67% / 100-0012
Cholecystokinin, CCK1 (CCKA) / Mouse / Pancreas / [125I]CCK-8 (0.02) / 3.17E-11 / CCK-8(sulfated) / 5.16E-11 / -5.67% / 100-0114
Cholecystokinin, CCK2 (CCKB) / Mouse / Pancreas / [125I]CCK-8 (0.02) / 0.2E-9 / CCK-8(sulfated) / 4.55E-10 / 6.87% / 100-0115
Endothelin, ET-A (h) / Human / Neuroblastoma / [125I]Endothelin-1 (0.033) / 1.6E-10 / Endothelin-1 / 1.74E-10 / 21.53% / 100-0085
Endothelin, ET-B (h) / Human / Astrocytoma / [125I]Endothelin-1 (0.025) / 2.1E-10 / Endothelin-1 / 2.44E-10 / 8.71% / 100-0124
Galanin,
Non-selective / Rat / Brain / [125I]Galanin (porcine) (100.0) / 1.0E-10 / Galanin (porcine) / 4.38E-10 / 11.6% / 100-0130
Neurokinin,
NK1 (NKA) (hr) / Rat / Submaxillary gland / [3H]Substance P (1.4) / 1.4E-9 / Substance P / 1.13E-8 / -9.65% / 100-0044
Neurokinin,
NK2 (NKA) (hr) / Human / Expressed in rCHO cell line / [125I]-NKA (0.1) / 5E-10 / Neurokinin A / 6.06E-10 / -0.18% / 100-0140
Neurokinin,
NK3 (NKB) / Rat / Cortex / [125I]-Eledoisin (0.1) / 1.5E-9 / Eledoisin / 5.06E-9 / 19.84% / 100-0141
Vasoactive Intestinal Peptide, non-selective / Rat / Forebrain / [125I]VIP (0.05) / 1.0E-9 / VIP / 1.23E-9 / 2.63% / 100-0170
Vasopressin 1 / Rat / Liver / [3H]Vasopressin-1 antagonist (0.5) / 0.3E-9 / Arg8-Vasopressin / 4.62E-9 / -15.19% / 100-0171

Enzyme / Species / Tissue / Substrate
(Concentration used
in the assay) / Km (M) / Reference compound / Ki (M) of reference compound / % inhibition of specific activity by
l-stepholidine (10E-7M)* / Novascreen
Catalog No.
Enzymes
Choline Acetyltransferase / Rat / Striatum / [14C]Acetyl Coenzyme A (50M) / 1.5E-6 / Bromoacetylcholine / 1.06E-7 / 15.51% / 200-0176
Esterase, Acetylcholine / Rat / Brain / Acetylthiocholine (0.3mM) / 1.4E-4 / Eserine / 3.63E-7 / 6.38% / 200-0083
Glutamic Acid Decarboxylase / Rat / Striatum / [14C]Glutamic acid (4.0M) / 1.3E-6 / AminoOxy acetic acid / 5.94E-10 / 2.45% / 200-0180
Oxidase,
MAO-A, peripheral / Rat / Liver / [14C]-5HT (Serotonin) (50.0M) / 8.7E-9 / Ro41-1049 / 5.51E-10 / 12.09% / 200-0078
Oxidase,
MAO-B, peripheral / Rat / Liver / [14C]Phenylethylamine (10.0M) / 4.0E-8 / Ro 16-6491 HCl / 1.69E-8 / -17.22% / 200-0079