Home Neuronal Signaling 5-HT Receptor antagonist RS-127445

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RS-127445 5-HT Receptor antagonist

RS-127445 (MT500) is a selective 5-HT2B receptor antagonist with pKi of 9.5 and pIC50 of 10.4, exhibits >1000-fold selectivity against other 5-HT receptors.

RS-127445 5-HT Receptor antagonist Chemical Structure

Chemical Structure

Molecular Weight: 281.33

Purity & Quality Control

Batch: S269801 DMSO]56 mg/mL]false]Ethanol]8 mg/mL]false]Water]Insoluble]false Purity: 98.72%
98.72

Related Products

Signaling Pathway

5-HT Receptor Signaling Pathways

5-HT Receptor Signaling Pathway

Mechanism of Action

Targets
5-HT2B [1] 5-HT2B [1]
10.4(pIC50) 9.5(pKi)

In vitro
In vitro RS-127445 is a novel high affinity, selective 5-HT2B receptor antagonist devoid of detectable intrinsic activity. This compound is found to have nM affinity and 1000 fold selectivity for the 5-HT2B receptor. It is thus among the highest affinity, most selective 5- HT2B receptor ligands. This chemical potently blocks the 5-HT evoked increase in inositol phosphate formation and blocks the 5-HT evoked increases in intracellular calcium concentrations with a potency 1000 times greater than that of yohimbine. [1]
Kinase Assay Radioligand binding
The selectivity of RS-127445 for 5-HT2B receptors is examined by testing this compound for affinity at over 100 additional ion channel or receptor binding sites. CHO-K1 cells expressing human 5-HT2A, 5-HT2B or 5-HT2C receptors are harvested using 2 mM EDTA in phosphate buffered saline. Cell membranes are prepared by four cycles of homogenization and centrifugation (48,000×g for 15 min). Each assay is established so as to achieve steady state conditions and to optimize specific binding. For the 5-HT2A receptor, membranes from 1×106 cells are incubated with 0.2 nM [3 H]-ketanserin at 32 °C for 60 min. Nonspecific binding is determined using 10 μM methysergide. For the 5-HT2B receptor, membranes from 1.5×106 cells are incubated with 0.2 nM [3 H]-5-HT at 48 °C for 120 min. Nonspecific binding is determined using 10 μM 5-HT. For the 5-HT2Creceptor, membranes from 3×10 5 cells are incubated with 0.5 nM [3 H]-mesuler -gine at 32 °C for 60 min. Nonspecific binding is determined using 10μM methysergide. Assays are terminated by vacuum filtration through glass fibre filters(GF/B) which has been pretreated with 0.1% polyethyleneimine. Total and bound radioactivity is determined by liquid scintillation counting. Greater than 90% specific binding is achieved in each of these assays.
Cell Research Cell lines HEK-293 cells expressing the human 5-HT2B receptor
Concentrations 10 μM
Incubation Time 20 min
Method

RS-127445, vehicle or other antagonists are pre-incubated with 240 μl of HEK-293 cells expressing the human 5-HT2B receptor suspension at 37 °C for 20 min. HEK-293 cells are incubated with[3H]-myoinositol (1.67 μCi/ml) in 162 cm2 flasks overnight at 37 °C in an inositol free Ham's F12 medium containing 10% dialyzed foetal bovine serum. The cells are harvested, washed five times with phosphate bufffered saline and resuspended in inositol free Ham's F12 media at density of approximately 3×103 cells/ml. The reactions are initiated by addition of 5-HT. Sixty minutes later, the reactions are terminated by adding 50 μl of ice-cold 20% perchloric acid, chilled in an ice-water bath for 10 min and then neutralized with 160μl of 1 N KOH. Each sample is diluted with 2 ml of 50 mM Tris-HCl, pH 7.4 at room temperature. The aqueous portion (2.2 ml) is transferred onto Dowex AG1X8 columns (1 ml, 1 : 1, w/v) which has been washed with 5 ml of distilled water. The columns are then washed with 18 ml of distilled water and the inositol phosphates are eluted with 3 ml of 1 N HCl. The eluted radioactivity is determined by liquid scintillation spectroscopy using a Packard 1900CA analyzer. [1]

In Vivo
In vivo RS-127445 is readily absorbed with no obvious dose or route-dependent limitations and rapidly absorbed following both oral and intraperitoneal administration with peak plasma concentrations being achieved within 15 min of dosing. This compound's concentration in the plasma are proportional to the administered dose. When administered at a dose of 5 mg/kg with approximately 60% of an intraperitoneal dose and 14% of the oral dose is bioavailable. Its concentration in the plasma is predicted to fully saturate accessible 5-HT2B receptors can be readily achieved and maintained in the rat. This chemical administered at 1 to 10 mg/kg with oral significantly inhibits visceral hypersensitivity up to 35 to 74% provoked by restraint stress. Oral administration produces a significant suppression of TNBS-induced visceral hypersensitivity (15 to 62% inhibition at 3 to 30 mg/ kg), although it has no significant effect on the visceral nociceptive threshold of native rats. It administrated orally with 1 to 30 mg/kg also dose -dependently reduce the restraint stress-induced defecation in native and TNBS-treated rats. [2]. This compound inhibits colonic motility and defecation. [3]
Animal Research Animal Models rats
Dosages 5 mg/kg
Administration Oral for 2.5 h ,intraperitoneal and intravenousroutes for 0.08 h

References
  • https://pubmed.ncbi.nlm.nih.gov/10455251/
  • http://www.ncbi.nlm.nih.gov/pubmed/ 19740115
  • https://pubmed.ncbi.nlm.nih.gov/19371340/

Chemical Information

Molecular Weight 281.33 Formula

C17H16FN3
CAS No. 199864-87-4 SDF Download SDF
Synonyms MT500
Smiles CC(C)C1=NC(=NC(=C1)C2=CC=C(C3=CC=CC=C32)F)N

Storage and Stability

Storage (From the date of receipt)

In vitro
Batch:

DMSO : 56 mg/mL ( (199.05 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Ethanol : 8 mg/mL

Water : Insoluble


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In vivo
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