| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| Other Sizes |
|
Purity: =100%
E55888 is a novel and potent agonist at the 5HT7 serotonin receptor
| Targets |
5-HT7 receptors (Ki = 2.5nM); 5-HT1A (Ki = 700nM)
|
|---|---|
| ln Vitro |
For the 5-HT(7) receptor agonists used, binding profile and intrinsic efficacy to stimulate cAMP formation in HEK-293F cells expressing the human 5-HT(7) receptor were also evaluated. AS-19 and E-55888 were selective for 5-HT(7) receptors. E-55888 was a full agonist whereas AS-19 and MSD-5a behaved as partial agonists, with maximal effects corresponding to 77% and 61%, respectively, of the cAMP response evoked by the full agonist 5-HT[1].
|
| ln Vivo |
In vivo results revealed that systemic administration of 5-HT(7) receptor agonists exerted a clear-cut dose-dependent antinociceptive effect that was prevented by 5-HT(7) receptor antagonists, but not by the 5-HT(1A) receptor antagonist. The order of efficacy (E-55888>AS-19>MSD-5a) matched their in vitro efficacy as 5-HT(7) receptor agonists. Contrary to agonists, a dose-dependent promotion of mechanical hypersensitivity was observed after administration of 5-HT(7) receptor antagonists, substantiating the involvement of the 5-HT(7) receptor in the control of capsaicin-induced mechanical hypersensitivity. These findings suggest that serotonin exerts an inhibitory role in the control of nociception through activation of 5-HT(7) receptors, and point to a new potential therapeutic use of 5-HT(7) receptor agonists in the field of analgesia[1].
|
| Enzyme Assay |
Binding affinities of E-55888 and AS-19 were determined through commercial radioligand binding assays by MDS Pharma and/or CEREP, as indicated, according to their standard assay protocols. Assays were done twice and concentrations were tested in duplicate. (For details regarding the specific receptors and experimental conditions see http://discovery.mdsps.com/Catalog/OnlineCatalog/Profiling/Assays/AssayList.aspx?id=5 and http://www.cerep.fr/Cerep/Users/pages/catalog/binding/catalog.asp).
For the determination of efficacy/potency of 5-HT7 receptor agonists used in this study, cAMP measurements were performed using a system based on homogeneous time-resolved fluorescence (HTRF) applied to human embryonic kidney (HEK)-293F cells that stably express the human 5-HT7(a) receptor, as previously described. This assay is based on competition between cAMP produced by cells and cAMP-XL665 conjugate for binding onto monoclonal anti-cAMP-cryptate conjugate. The HTRF cAMP kit was used according to the instructions of the manufacturer. Briefly, after overnight incubation in serum-free medium, cells were added to 96-well plates (20,000 cells/well) in Ham’s F12 incubation buffer (40μl/well) containing 1mM 3-isobutyl-1-methyl-xanthine (IBMX) and 20μM pargyline. Then, 10μl of different concentrations of the tested compound was added, and the plates were incubated for 30min at 37°C. The reaction was stopped by using a mixture of 25μl of cryptate and 25μl of XL-665 prepared in the lysis buffer supplied by the manufacturer. Plates were then incubated for an additional hour at room temperature, and cAMP contents were calculated from the 665nm/620nm ratio using a RubyStar Plate reader [1]. |
| Cell Assay |
E-55888 is a new highly selective, potent 5-HT7 receptor agonist with high efficacy. It showed high affinity for 5-HT7 receptors (Ki=2.5nM) with significant affinity for 5-HT1A (Ki=700nM) and no significant affinity (Ki>1μM or% inhibition at 10−6M lower than 50%) for other 5-HT receptor subtypes and 170 additional targets including receptors, transporters and ion channels (in vitro binding screening packages by CEREP and MDS Pharma Services) (Table 1). When tested in a functional assay, E-55888 concentration-dependently increased cAMP formation in HEK-293F/h5-HT7 cells and behaved as a full agonist, with efficacy and potency (Emax=99±1% and EC50=16±1nM) similar to those of 5-HT (Emax=100 and EC50=11nM). [1]
This compound/AS-19 was also tested in a functional assay (cAMP stimulation in HEK-293F/h5-HT7 cells) and was found to behave as a potent (EC50=9±1nM) but partial 5-HT7 receptor agonist, with a maximal effect reaching 77% of that of 5-HT[1] |
| Animal Protocol |
Male CD1 mice aged from 6 to 8 weeks old were used. Animals were housed in groups of five, provided with food and water ad libitum and kept under controlled laboratory conditions (temperature: 21±1°C; 12h light/dark cycles with light on at 07:00h). Experiments were carried out in a soundproof and air-regulated room. The number of mice ranged from 9 to 16 in each experimental group as 3–4 independent experiments using 3–4 mice per group were always performed. All experimental procedures and animal husbandry were conducted according to ethical principles for the evaluation of pain in conscious animals and to ethical guidelines of the European Community on the Care and Use of Laboratory Animals (European Communities Council Directive of 24 November 1986, 86/609/ECC), and received approval by the Local Ethical Committee.[1]
When fixed at 1g, the pressure exerted by the filament is insufficient to induce timely pain responses in normal control mice (in the absence of capsaicin), and thus prompt withdrawal responses found 15min after sensitization with 1μg capsaicin reflect mechanical hypersensitivity (see Section 3 and Fig. 2). This approach enabled us to evaluate the possible antinociceptive effect of 5-HT7 and 5-HT1A receptor ligands. |
| References |
[1]. 5-HT7 receptor activation inhibits mechanical hypersensitivity secondary to capsaicin sensitization in mice. Pain. 2009 Feb;141(3):239-47. https://pubmed.ncbi.nlm.nih.gov/19118950/
|
| Additional Infomation |
This work aimed to evaluate the potential role of the 5-HT(7) receptor in nociception secondary to a sensitizing stimulus in mice. For this purpose, the effects of relevant ligands (5-HT(7) receptor agonists: AS-19, MSD-5a, E-55888; 5-HT(7) receptor antagonists: SB-258719, SB-269970; 5-HT(1A) receptor agonist: F-13640; 5-HT(1A) receptor antagonist: WAY-100635) were assessed on capsaicin-induced mechanical hypersensitivity, a pain behavior involving hypersensitivity of dorsal horn neurons (central sensitization). For the 5-HT(7) receptor agonists used, binding profile and intrinsic efficacy to stimulate cAMP formation in HEK-293F cells expressing the human 5-HT(7) receptor were also evaluated. AS-19 and E-55888 were selective for 5-HT(7) receptors. E-55888 was a full agonist whereas AS-19 and MSD-5a behaved as partial agonists, with maximal effects corresponding to 77% and 61%, respectively, of the cAMP response evoked by the full agonist 5-HT. Our in vivo results revealed that systemic administration of 5-HT(7) receptor agonists exerted a clear-cut dose-dependent antinociceptive effect that was prevented by 5-HT(7) receptor antagonists, but not by the 5-HT(1A) receptor antagonist. The order of efficacy (E-55888>AS-19>MSD-5a) matched their in vitro efficacy as 5-HT(7) receptor agonists. Contrary to agonists, a dose-dependent promotion of mechanical hypersensitivity was observed after administration of 5-HT(7) receptor antagonists, substantiating the involvement of the 5-HT(7) receptor in the control of capsaicin-induced mechanical hypersensitivity. These findings suggest that serotonin exerts an inhibitory role in the control of nociception through activation of 5-HT(7) receptors, and point to a new potential therapeutic use of 5-HT(7) receptor agonists in the field of analgesia.[1]
|
| Molecular Formula |
C16H23N3
|
|---|---|
| Molecular Weight |
257.373923540115
|
| Exact Mass |
257.1892
|
| Elemental Analysis |
C, 74.67; H, 9.01; N, 16.33
|
| CAS # |
1034142-33-0
|
| PubChem CID |
24825775
|
| Appearance |
Solid powder if stored under nitrogen but this product is extremely hydroscopic and can become oily (ointment) when exposed to air
|
| LogP |
3
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
2
|
| Rotatable Bond Count |
4
|
| Heavy Atom Count |
19
|
| Complexity |
279
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
N(C)(C)CCC1=CC=CC(C2=C(C)N(C)N=C2C)=C1
|
| InChi Key |
MFUWRMRKXKCSPL-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C16H23N3/c1-12-16(13(2)19(5)17-12)15-8-6-7-14(11-15)9-10-18(3)4/h6-8,11H,9-10H2,1-5H3
|
| Chemical Name |
N,N-dimethyl-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenethylamine
|
| Synonyms |
E-55888; E 55888; 1034142-33-0; E55888; E-55888; CHEMBL4205349; N,N-dimethyl-2-[3-(1,3,5-trimethylpyrazol-4-yl)phenyl]ethanamine; N,N-dimethyl-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenethylamine; Benzeneethanamine, N,N-dimethyl-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)-; dimethyl-{2-[3-(1,3,5-trimethyl-1H-pyrazol-4-yl)-phenyl]-ethyl}-amine; E55888
|
| HS Tariff Code |
2934.99.9001
|
| 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, as this product is extremely hydroscopic and can become oily (ointment) when exposed to air. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
|
|---|---|
| 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 | 3.8855 mL | 19.4273 mL | 38.8546 mL | |
| 5 mM | 0.7771 mL | 3.8855 mL | 7.7709 mL | |
| 10 mM | 0.3885 mL | 1.9427 mL | 3.8855 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA