5-HT7 Receptor (pKi = 7.5)
SB 258719 targets the 5-HT7 receptor, a G protein-coupled receptor that is activated by serotonin (5-hydroxytryptamine, 5-HT). The 5-HT7 receptor is widely expressed in the central nervous system, particularly in regions involved in mood regulation, cognition, and circadian rhythm. It is also expressed in peripheral tissues, including the gastrointestinal tract and vasculature. SB 258719 is a selective antagonist of the 5-HT7 receptor with high affinity (pKi = 7.5). By blocking 5-HT7 receptor activation, SB 258719 can be used to study the physiological and pathological roles of this receptor. The compound's selectivity for 5-HT7 over other serotonin receptors makes it a valuable tool for dissecting 5-HT7-specific functions.

Without significantly altering the peak response to 5-CT, SB 258719 (1, 3 and 10 μM; HEK 293 cells) causes a concentration-related rightward shift in the 5-CT concentration response curve [1].

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SB 258719 demonstrates potent in vitro activity as a 5-HT7 receptor antagonist with high affinity (pKi = 7.5). In receptor binding assays, SB 258719 selectively binds to the 5-HT7 receptor with high affinity. In functional assays, the compound antagonizes 5-HT7 receptor-mediated signaling, blocking the effects of serotonin at this receptor. The compound's selectivity for 5-HT7 over other serotonin receptors and other receptor classes makes it a valuable tool for studying 5-HT7-specific signaling pathways. SB 258719 has been used in research on cancer and neurological diseases, highlighting its utility in diverse research areas.

The 5-CT-induced hypothermia is greatly attenuated by SB 258719 (5–20 mg/kg; i.p.) [2].

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SB 258719 has been used in preclinical studies to investigate the role of 5-HT7 receptors in various physiological and pathological processes. The compound's high affinity and selectivity for 5-HT7 make it a valuable tool for in vivo studies. SB 258719 can be used for the research of cancer and neurological disease. In animal models, 5-HT7 receptor antagonists have been shown to modulate mood, cognition, and pain perception. SB 258719 has been used to study the role of 5-HT7 receptors in these processes. Detailed in vivo efficacy data, including specific model results and dosing regimens, are available in the primary literature. The compound's utility in studying 5-HT7 receptor function supports its value in preclinical research.

Measurement of [35S]-GTPgS binding [ 35S]-GTPgS binding to 5-HT7(a)/HEK293 membranes was measured using the method of Thomas et al. (1995a). Brie¯y, membranes (20 ± 30 mg protein) were preincubated (308C for 30 min) in 20 mM HEPES bu€er (pH 7.4) in the presence of 3 mM MgCl2, 100 mM NaCl, 10 mM GDP, 0.2 mM ascorbate and in the presence or absence of test drugs. Incubations (30 min, 308C) were started by addition of [ 35S]-GTPgS (0.1 ± 3 nM) followed by vigorous mixing and stopped by rapid ®ltration through Whatman GF/B ®lters followed by ®ve 1 ml washes with ice-cold bu€er containing 20 mM HEPES and 3 mM MgCl2. All determinations within an experiment were performed in duplicate. Radioactivity on the ®lters was determined using liquid scintillation spectrometry.[1]

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The in vitro receptor binding assay for SB 258719 measures the binding affinity of the compound to the 5-HT7 receptor. Membrane preparations from cells expressing the human 5-HT7 receptor are incubated with varying concentrations of SB 258719 (typically ranging from nanomolar to micromolar) in the presence of a radiolabeled 5-HT7 receptor ligand (e.g., 3H-5-HT or 3H-LSD). The amount of bound radioligand is measured by scintillation counting, and Ki values are determined by fitting competition binding curves. The pKi value of 7.5 indicates high affinity binding. The compound is dissolved in DMSO and diluted in assay buffer to achieve the desired final concentrations, with DMSO concentration kept constant across all wells. Appropriate positive controls (known 5-HT7 antagonists) and negative controls (DMSO vehicle) are included in each assay run. Selectivity is assessed by testing the compound against other serotonin receptors and other receptor classes.

HEK293 cells stably expressing the human 5-HT7(a) receptor were grown in Minimum Essential Medium (MEM) containing 10% dialysed foetal calf serum, G418 sulphate (1 mM), glutamine (2 mM) and non-essential amino acids (1%). Cells were grown to con¯uence, washed with phosphate bu€ered saline (PBS), and pelleted by centrifugation (1000 g) in PBS containing EDTA (0.1 mM) and dithiothreitol (1 mM). Pellets were stored at 7808C prior to membrane preparation. For preparation of membranes, cell pellets were washed twice by homogenisation (Polytron, 15 s, setting 5) and centrifugation (50,000 g, 15 min, 48C) in 20 volumes of Tris HCl (25 mM pH 7.4) containing EDTA (0.1 mM). Membranes were then resuspended in bu€er and incubated (378C, 20 min). Following centrifugation and a further wash at 48C, the membranes were ®nally re-suspended at a membrane concentration equivalent to 2.56107 cells ml71 and stored at 7808C prior to use[1].

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The in vitro cellular assay for SB 258719 is performed using cells expressing the 5-HT7 receptor, such as HEK293 cells stably transfected with 5-HT7R. Cells are cultured in appropriate medium and treated with varying concentrations of SB 258719 or vehicle control (DMSO). Cells are stimulated with serotonin to activate the 5-HT7 receptor, and receptor activation is measured by assessing downstream signaling events such as cAMP accumulation (5-HT7 is a Gs-coupled receptor that increases cAMP). The inhibition of serotonin-induced cAMP accumulation by SB 258719 is quantified, and IC50 or KB values are determined by fitting dose-response curves. The compound's selectivity for 5-HT7 over other serotonin receptors can be assessed in similar assays using cells expressing other 5-HT receptor subtypes.

Animal/Disease Models: Male Swiss Webster mouse (20-25 g) [2].
Doses: 5-20 mg/kg
Route of Administration: intraperitoneal (ip) injection
Experimental Results: Dramatically attenuated hypothermia induced by 5-CT.

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In vivo animal experiments with SB 258719 are typically conducted using rodent models of neurological or psychiatric disorders. SB 258719 is administered via intraperitoneal injection or oral gavage at various doses. Behavioral tests relevant to the condition being studied (e.g., forced swim test for depression, elevated plus maze for anxiety, novel object recognition for cognition) are performed. The effects of SB 258719 on behavior are compared to vehicle-treated controls. Additionally, the compound's effects on 5-HT7 receptor-mediated physiological responses can be assessed. At the end of the study, brain tissues may be harvested for analysis of receptor occupancy and downstream signaling markers. Detailed experimental protocols, including dosing regimens and endpoints, are described in the primary literature.

Detailed pharmacokinetic (PK) parameters for SB 258719 are not extensively documented in publicly available sources. As a small-molecule 5-HT7 antagonist with a molecular weight of 338.51, the compound is expected to have reasonable oral bioavailability and good central nervous system penetration. SB 258719 has a chemical formula of C18H30N2O2S. The compound is soluble in DMSO for formulation purposes. For in vivo administration, the compound is typically formulated using appropriate vehicles to ensure adequate solubility and stability. The compound should be stored as a powder at -20°C (stable for up to 3 years) or at 4°C (stable for up to 2 years). Detailed PK parameters including half-life, clearance, volume of distribution, and maximum concentration (Cmax) are available in the primary literature and should be consulted for specific experimental planning.

Comprehensive toxicological data for SB 258719 are not extensively documented in publicly available sources. As a research-grade compound, SB 258719 is intended for laboratory research purposes only and is not approved for human therapeutic use. Standard laboratory safety practices should be followed when handling this compound, including the use of appropriate personal protective equipment and working in a well-ventilated area. The compound should be stored according to the manufacturer's recommendations to maintain stability and prevent degradation. In animal studies, SB 258719 has been reported to be tolerated at the doses tested for behavioral studies. However, comprehensive toxicological profiling (e.g., LD50, maximum tolerated dose, organ-specific toxicity) is not available from the current search results and would require consultation of the primary literature or safety data sheets.

[1]. Functional characterisation of the human cloned 5-HT7 receptor (long form); antagonist profile of SB-258719. Br J Pharmacol. 1998;124(6):1300-1306.

[2]. The hypothermic effect of 5-CT in mice is mediated through the 5-HT7 receptor. Neuropharmacology. 2003;44(8):1031-1037.

[3]. 5-Hydroxytryptamine promotes hepatocellular carcinoma proliferation by influencing β-catenin. Mol Oncol. 2016;10(2):195-212.

1. This study investigated the functional characteristics of the long human clone 5-HT7 receptor (named h5-HT7(a)) using various 5-HT receptor agonists and antagonists, and compared them with their binding properties. Receptor function was assessed by measuring adenylate cyclase activity in the washed membrane of HEK293 cells stably expressing the recombinant h5-HT7(a) receptor. 2. The receptor binding properties, measured by competition with [3H]-5-CT, were consistent with previously reported characteristics of the h5-HT7(a) receptor. The selective 5-HT7 receptor antagonist SB-258719 ((R)-3,N-dimethyl-N-[1-methyl-3-(4-methylpiperidin-1-yl)propyl]benzenesulfonamide) exhibited high affinity for this receptor (pKi 7.5). 3. In the adenylate cyclase function assay, both 5-CT and 8-OH-DPAT were complete agonists compared to 5-HT, and the power order of the agonists (5-CT > 5-HT > 8-OH-DPAT) was consistent in both functional and binding studies. 4. Risperidone, methapine, mesuermoeline, clozapine, olanzapine, ketoserin, and SB-258719 all antagonized 5-CT-stimulated adenylate cyclase activity. Schild analysis of the antagonistic effect of SB-258719 showed a pA2 value of 7.2 ± 0.2, with no significant difference in slope compared to 1, consistent with competitive antagonism. 5. These antagonists also inhibited basal adenylate cyclase activity, and their power order was consistent with the order of antagonist power and binding affinity. Compared to other tested antagonists, SB-258719 and mesuermoeline both exhibited significant partially inverse agonist characteristics. The inhibitory effects of these antagonists appear to be mediated by the 5-HT7 receptor and reflect inverse agonist activity. 6. In conclusion, the h5-HT7(a) receptor in this expression system exhibits the expected binding and functional properties and constitutive activity, revealing inverse agonist activity of a range of antagonists. [1]

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SB 258719 is a research compound developed for studying the role of 5-HT7 receptors in neurological and psychiatric disorders. The compound is a selective 5-HT7 receptor antagonist with high affinity (pKi = 7.5). SB 258719 can be used for the research of cancer and neurological disease. The compound's selectivity for 5-HT7 over other serotonin receptors makes it a valuable tool for dissecting 5-HT7-specific functions. SB 258719 is not currently in clinical trials nor approved for therapeutic use; it remains an investigational tool compound for preclinical research. The compound is available from various chemical suppliers for research purposes. Its utility lies in its ability to selectively antagonize 5-HT7 receptors and probe their role in mood, cognition, and other physiological processes.

C18H30N2O2S

338.5080037117

338.202799

195199-95-2

SB 258719 hydrochloride;1217674-10-6

5312148

Light yellow to yellow liquid

4.946

0

4

6

23

452

1

S(C1=CC=CC(C)=C1)(N(C)[C@H](C)CCN1CCC(C)CC1)(=O)=O

UIZKHTBWJSUGOV-UNTBIKODSA-N

InChI=1S/C18H30N2O2S.ClH/c1-15-8-11-20(12-9-15)13-10-17(3)19(4)23(21,22)18-7-5-6-16(2)14-18;/h5-7,14-15,17H,8-13H2,1-4H3;1H/t17-;/m1./s1

N,3-dimethyl-N-[(2R)-4-(4-methylpiperidin-1-yl)butan-2-yl]benzenesulfonamide Hydrochloride

SB-258719 HCl; SB258719; SB258719; SB 258719; SB-258719; GF43CP5LXQ; SB258719; N,3-dimethyl-N-[(2R)-4-(4-methylpiperidin-1-yl)butan-2-yl]benzenesulfonamide; UNII-GF43CP5LXQ; CHEMBL12264; SB 258719 HCl.

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~250 mg/mL (~738.53 mM)

Solubility in Formulation 1: ≥ 6.25 mg/mL (18.46 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 62.5 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 6.25 mg/mL (18.46 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 62.5 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 6.25 mg/mL (18.46 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 62.5 mg/mL clear DMSO stock solution to 900 μL corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)