|
Biological Activity
|
|
Description
|
JTC-801 is a selective opioid receptor-like1 (ORL1) receptor antagonist with IC50 of 94 nM. |
|
Targets
|
Opioid receptor-like1 (ORL1) |
|
|
|
|
|
|
IC50 |
94 nM [1] |
|
|
|
|
|
|
In Vitro
|
JTC-801 displays about 12.5-, 129-, and 1055-fold selectivity for ORL1 receptor (Ki = 8.2 nM) over μ-, κ-, and δ-opioid receptors, respectively. JTC-801 does not inhibit forskolin-stimulated cyclic AMP accumulation in human ORL1 receptor-expressing HeLa cells, but it prevents nociceptin-induced inhibition of cyclic AMP accumulation, indicating that JTC-801 possesses full antagonistic activity. [2] In rat cerebrocortical membrane, JTC-801 inhibits ORL1 receptor with IC50 of 472 nM and μ-receptor with IC50 of 1831 nM. JTC-801 completely antagonizes the suppression of nociceptin on forskolin-induced accumulation of cyclic AMP with IC50 of 2.58 μM in HeLa cells expressing ORL1 receptor. [1] |
|
In Vivo
|
Oral administration of JTC-801 (0.3-3 mg/kg) antagonizes nociceptin-induced allodynia in mice, and shows analgesic effect in a hot plate test using mice and in a formalin test using rats. [2] In mouse hot-plate test, JTC-801 prolongs escape response latency (ERL) or exposed heat stimulus with minimum effective doses (MED) of 0.01 mg/kg by i.v. or 1 mg/kg by p.o. In the rat formalin test, JTC-801 reduces both the first and second phases of the nociceptive response with MED of 0.01 mg/kg71 by i.v. or 1 mg/kg by p.o. [1] JTC-801 dose-dependently normalizes paw withdrawal latency (PWL). Although JTC-801 does not inhibit a chronic constriction injury (CCI)-induced decrease in bone mineral content (BMC) and bone mineral density (BMD), it inhibits an increase in the number of osteoclasts. [3] Tactile allodynia induced by L5/L6 spinal nerve ligation is reversed by both systemic (3-30 mg/kg) and spinal (22.5 and 45 pg) JTC-801 in a dose-dependent manner. Furthermore, systemic JTC-801 reduces Fos-like immunoreactivity in the dorsal horn of the spinal cord (laminae I/II). [4] JTC-801 produces dose-dependent mechanical and cold anti-allodynic effects with ED50 of 0.83 mg/kg and 1.02 mg/kg, respectively. [6] |
|
Clinical Trials
|
|
|
Features
|
|
|
Combination Therapy
|
|
Description
|
JTC-801 (1 mg/kg, i.p.) blocks a significant proportion of the hypothermia caused by WIN 55212-2 (2.5, 5, and 10 mg/kg, i.p.) or another cannabinoid agonist, CP-55940 (1 mg/kg, i.p.). [5] JTC-801 in combination with WIN 55212-2 produces greater anti-allodynic effects, compared with the dose-response curves of each drug alone. [6] |
|
Protocol
|
|
Kinase Assay
[1]
|
| Human ORL1 receptor binding affinity |
Human ORL1 receptor expressed in HeLa cells are harvested and homogenized in 50 mM Tris buffer (pH 7.4) containing 1 mM EDTA. After centrifugation for 30 minutes at 40,000×g, the pellets are resuspended in buffer containing 50 mM Tris, supplemented with 10 mM MgCl2 and 2 mM EGTA, and used as membrane preparations. 50 mM Tris (pH 7.4) supplemented with 2 mM EDTA and 0.1 mM (p-amidoinophenyl) methanesulphonyl fluoride hydrochloride containing 0.2% bovine serum albumin is used for the binding buffer. For saturation binding assay, the cell membrane preparations are incubated for 60 minutes at 24 °C with various concentrations of [3H]-nociceptin. Nonspecific binding is determined in the presence of 1 mM unlabelled nociceptin. For competitive assay, the cell membrane preparations (4.17 μg/well) are incubated for 60 minutes at 24 °C with 50 pM [3H]-nociceptin in the presence of various concentrations of JTC-801 (10 nM-10 μM). JTC-801 is dissolved in DMSO and diluted in binding buffer, and then added to the incubation mixture. Final concentration of vehicle is 1% DMSO in binding buffer. After incubation for 60 minutes, the membrane preparations are rapidly filtrated over Whatman GF/B glass filters which are pretreated with 0.1% polyethyleneimine, and the radioactivity on each filter is measured by liquid scintillation counting. IC50 value is calculated as the concentration of JTC-801 required to displace 50% inhibition of the [3H]-nociceptin. |
|
Animal Study
[1]
|
| Animal Models |
Male ICR (CD-1) subjected to nociceptin-induced allodynia test or hot plate test, and Male SD rats subjected to formalin-induced paw-licking response |
| Formulation |
Suspended in 0.5% methyl cellulose solution or dissolved in 5% sorbitol |
| Doses |
~10 mg/kg |
| Administration |
Orally or injected i.v. |
|
References |
|
[1] Yamada H, et al. Br J Pharmacol, 2002, 135(2), 323-332.
|
|
[2] Shinkai H, et al. J Med Chem, 2000, 43(24), 4667-4677.
|
|
[3] Suyama H, et al. Neurosci Lett, 2003, 351(3), 133-136.
|
|
[4] Tamai H, et al. Eur J Pharmacol, 2005, 510(3), 223-228.
|
|
[5] Rawls SM, et al. Neuropeptides, 2007, 41(4), 239-247.
|
|
[6] Gunduz O, et al. Pharmacol Biochem Behav, 2011, 99(4), 540-544.
|
|