| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
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| Targets |
5-HT Receptors (5-HT4; 5-HT3; 5-HT2A; 5-HT2C)
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| ln Vitro |
1. The 5-HT4 receptor has only recently been identified but has yet to be cloned. This paper describes the pharmacology of a potent and selective 5-HT4 receptor antagonist, GR113808, which will be useful in the further characterization of this receptor. 2. On the guinea-pig ascending colon, GR113808 (1 nM-0.1 microM) behaved as an antagonist of 5-hydroxytryptamine (5-HT)-induced contraction, producing rightward displacements of the concentration-effect curve to 5-HT and a concentration-related depression of the maximum effect. However, the compound had no effect on cholecystokinin (CCK-8)-induced contraction in concentrations up to 1 microM. 3. In the guinea-pig colon preparation, onset and offset of the antagonism by GR113808 of 5-HT-induced contraction was examined. Incubation of the tissues for either 15 min, 30 min or 60 min produced similar rightward displacements of the concentration-effect curves to 5-HT, with no increase in the degree of depression of the maxima with increasing time of incubation. Experiments examining offset of antagonism (0.01 microM) demonstrated that washout for 30 min was required to reverse fully the effects of the antagonist. 4. Potency estimates in the colon for GR113808 were made by determining approximate pA2 values (30 min) using the Gaddum equation. The values obtained were 9.2, 9.7 and 9.2 when tested against the agonists 5-HT, 5-methoxytryptamine and R,S-zacopride respectively. 5. On the carbachol-contracted tunica muscularis mucosae preparation of the rat thoracic oesophagus, GR113808 behaved as an antagonist of 5-HT-induced relaxation, producing no reduction in maximum response [1].
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| ln Vivo |
GR113808 (intraperitoneal injection; 1 mg/kg; 5-7 days starting 24 hours after conduction inflammation) revealed a considerable rise in the active disease index (DAI), which takes into account the presence of matrix groundwater, weight loss, and matrix consistency [2].
Rectal administration of tegaserod reduced the severity of colitis, compared to mice given vehicle, and accelerated recovery from active colitis. Rectal tegaserod did not improve colitis in 5-HT4R knockout mice, and intraperitoneally administered tegaserod did not protect wild-type mice from colitis. Tegaserod increased proliferation of crypt epithelial cells. Stimulation of 5-HT4R increased Caco-2 cell migration and reduced oxidative stress-induced apoptosis; these actions were blocked by co-administration of the 5-HT4R antagonist GR113808. In non-inflamed colons of wild-type mice not receiving tegaserod, inhibition of 5-HT4Rs resulted in signs of colitis within 3 days. In these mice, epithelial proliferation decreased and bacterial translocation to the liver and spleen was detected. Daily administration of tegaserod increased motility in inflamed colons of guinea pigs and mice, whereas administration of GR113808 disrupted motility in animals without colitis.
Conclusions
5-HT4R activation maintains motility in healthy colons of mice and guinea pigs reduces inflammation in colons of mice with colitis. Agonists might be developed as treatments for patients with inflammatory bowel diseases [2].
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| Cell Assay |
Cell lines culture and treatment for oxidative stress [2]
Human Caco-2 cells, a epithelial colorectal adenocarcinoma cell line (ATCC, UK), were maintained in a humidified atmosphere of 95% air and 5% CO2 at 37°C in Dulbecco’s modified Eagle’s medium (DMEM), supplemented with 10% (v/v) fetal bovine serum (FBS), 100 U/mL penicillin and 100 Ag/mL streptomycin. To produce oxidative stress in Caco-2, cells were treated with 200 μM of H2O2 in phosphate-buffered saline (PBS) for 30 min19,18 24 h after the cells were seeded. The sulforhodamine B (SRB) assay was used to determine cell density, as previously described and validated. 20,19,18 Cells grown in 24 well plates were treated for 1 hr with vehicle, agonist or agonist plus antagonist. Supernatant was aspirated from wells and cells were fixed with cold trichloroacetic acid solution (30% w/v) at 4°C for 1 h. Fixed cells were washed with H2O and dried, and SRB solution (0.057% w/v) was applied to stain the cellular protein contents. Using spectrophotometry, absorbance was measured at 540 nm with a reference wavelength of 630 nm. Scratch Assay [2] Caco-2 cells (provided by Dr. J Turner, University of Chicago) were cultured in 6 well plates to ~90% confluence in DMEM high glucose supplemented with 10% FBS, 1% GlutaMAX, 10 mM Hepes and 100 U/mL Penicillin-Streptomycin. Once ~90% confluence was reached, three wounds were created using a sterile 200 uL pipet tip dragged perpendicular to a black line drawn on the underside of the plate for reference. Images were captured of each scratch at time points 0 h and 48 h with a Nikon D7100 camera on a Nikon Diaphot inverted microscope at 4X magnification. Only scratches whose edges could be captured in one frame at time point 0 h were included for final analysis. Measurements were taken from edge to edge at time 0 h and compared to measurements from 48 h using ImageJ software.21 The reported values are the difference between 0 h and 48 h, with higher values representing increased cellular migration. Three separate experiments were conducted with all three conditions. |
| Animal Protocol |
Animal/Disease Models: Mice [1]
Doses: 1 mg/Kg Route of Administration: intraperitoneal (ip) injection; 1 mg/kg; 5-7 days starting 24 hrs (hrs (hours)) after induction of colitis Experimental Results: Blocking the effects of tegaserod after induction of colitis Protective effects. Colitis Paradigms [2] For the prevention experiments, mice received DSS for 5 days and were then switched to tap water for 2 days. Alternatively, mice and guinea pigs were given a single enema of TNBS. Enemas with either vehicle (1% dimethyl sulfoxide (DMSO) in 0.9% saline; 0.2 mL/mouse) or drug (tegaserod and GR113808 both delivered at 1 mg/Kg) were administered daily for 5–7 days starting 24 h after induction of colitis. These doses were chosen because they were effective in previous studies of the effects of luminal administration of these compounds on visceral sensitivity.8 In a preliminary study, we found that enema treatment did not affect the histological damage score (HDS) (naïve, 0.6±0.25, n=5; vehicle enema, 1.1±0.3; p=0.13, n=7). In another preliminary study, involving enema administration of a vehicle solution containing 0.5% Evans Blue, we found that the solution delivered spread as far orally as the cecum after 10 minutes. Animals were euthanized on day 6 or 7 for TNBS or DSS studies, respectively. In the recovery paradigm drug treatment began on day 6 and lasted for 10 days, with animals euthanized on day 15. The time courses were chosen to test the effectiveness of the treatments leading up to the peak of inflammation, or beginning once the peak had been reached. Transcript levels for the 5-HT4 receptor were not altered in DSS (p=0.56) or TNBS (p=0.9) colitis. |
| References | |
| Additional Infomation |
GR 113808 is an indolyl carboxylate ester obtained by formal condensation between the carboxy group of 1-methylindole-3-carboxylic acid with the hydroxy group of N-{2-[4-(hydroxymethyl)piperidin-1-yl]ethyl}methanesulfonamide. It has a role as a serotonergic antagonist. It is an indolyl carboxylate ester, a member of piperidines and a sulfonamide.
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| Molecular Formula |
C19H27N3O4S
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| Molecular Weight |
393.5
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| Exact Mass |
393.172
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| Elemental Analysis |
C, 57.99; H, 6.92; N, 10.68; O, 16.26; S, 8.15
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| CAS # |
144625-51-4
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| PubChem CID |
119376
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| Appearance |
White to off-white solid powder
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| Density |
1.3g/cm3
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| Boiling Point |
572.3ºC at 760 mmHg
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| Flash Point |
299.9ºC
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| Vapour Pressure |
4.19E-13mmHg at 25°C
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| Index of Refraction |
1.613
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| LogP |
3.005
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
27
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| Complexity |
599
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
MOZPSIXKYJUTKI-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C19H27N3O4S/c1-21-13-17(16-5-3-4-6-18(16)21)19(23)26-14-15-7-10-22(11-8-15)12-9-20-27(2,24)25/h3-6,13,15,20H,7-12,14H2,1-2H3
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| Chemical Name |
[1-[2-(methanesulfonamido)ethyl]piperidin-4-yl]methyl 1-methylindole-3-carboxylate
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| Synonyms |
GR-113808; GR113808; 144625-51-4; GR-113,808; GR113,808; 1H-Indole-3-carboxylic acid, 1-methyl-, (1-(2-((methylsulfonyl)amino)ethyl)-4-piperidinyl)methyl ester; ZT350OYT3I; (1-(2-(Methylsulfonylamino)ethyl)-4-piperidinyl)methyl 1-methyl-1H-indole-3-carboxylate; CHEBI:73380; GR 113808
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~39 mg/mL (~99.11 mM)
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5413 mL | 12.7065 mL | 25.4130 mL | |
| 5 mM | 0.5083 mL | 2.5413 mL | 5.0826 mL | |
| 10 mM | 0.2541 mL | 1.2706 mL | 2.5413 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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