| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| Other Sizes |
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| Targets |
TP/thromboxane-prostanoid receptor
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|---|---|
| ln Vitro |
SQ29548 attenuates the decrease in the viability of SH-SY5Y cells induced by H2O2.[2]
SQ29548 reduces the intracellular ROS level in SH-SY5Y cells induced by H2O2.[2] SQ29548 ameliorates the decrease in SOD2 and catalase levels in SH-SY5Y cells induced by H2O2.[2] SQ29548 reduces the apoptosis of SH-SY5Y cells induced by H2O2.[2] SQ29548 inhibits cleaved caspase expression in SH-SY5Y cells induced by H2O2.[2] SQ29548 inhibits ERK and p38 signaling pathway activation in SH-SY5Y cells induced by H2O2.[2] |
| ln Vivo |
Serotonin-induced contraction of the MetS vessels was significantly inhibited in the presence of the selective PLA2 inhibitor quinacrine (10−6 M), the COX inhibitor indomethacin (10−5 M), and the TP receptor antagonist SQ29548 (10−6 M), respectively.[1]
Therefore, we evaluated the effects of SQ 29,548, a potent and selective TP antagonist-on neuromotor performance, neurodegeneration, reactive astrocytosis, and c-Fos protein immunoreactivity after pilocarpine-induced status epilepticus (SE) in mice. Adult C57BL/6 mice received intracerebroventricular SQ 29,548 injections 90 min and 24 h after pilocarpine-induced SE. We found that SQ 29,548 prevented the impairment of neuromotor performance (Neuroscore test) 48 h after pilocarpine-induced SE. Data analysis suggested the existence of two subgroups of SQ 29,548-treated post-SE animals. Eight out of 12 SQ 29,548-treated animals displayed Neuroscore values identical to those of vehicle-treated controls, and were considered SQ 29,548 responders. However, 4 out of 12 SQ 29,548-treated animals did not show any improvement in Neuroscore values, and were considered SQ 29,548 non-responders. Treatment with SQ 29,548 attenuated SE-induced increase in the number of FJC- or GFAP-positive cells in the hippocampus of SQ 29,548 responders. In addition, SQ 29,548 prevented the SE-elicited increase of c-Fos immunoreactivity in the hippocampus. In summary, our results suggest that the TP antagonist (SQ 29,548) improves neurological outcome after pilocarpine-induced SE in mice. The existence of SQ 29,548 responders and non-responders was suggested by results from the Neuroscore test. Additional studies are needed to understand the mechanisms underlying these findings, as well as the potential uses of TP antagonists in the treatment of seizure-induced damage.[4] |
| Enzyme Assay |
The binding of 5,6-3H(1S-[1 alpha, 2 beta(5Z), 3 beta, 4 alpha])-7-[3-([2-[(phenyl amino)carbonyl]hydrazino]methyl)-7-oxabicyclo[2.2.1]hept-2-yl]-5- heptenoic acid to receptors in human washed platelets (WP) and platelet membranes (PM) was characterized with regard to kinetics, saturability and competitive inhibition by putative thromboxane A2/prostaglandin H2 (TP)-receptor ligands. Specific binding of [3H]SQ 29,548 routinely amounted to 90 to 97% of total binding. The rate of association was 1.6 x 10(7) and 2.5 x 10(7) M-1 x min-1 in WP and PM, respectively. The corresponding rate of dissociation was 0.07 and 0.12 min-1, resulting in dissociation constants of 4.1 and 5.8 nM in WP and PM, respectively. Saturable binding to a single class of receptors indicated a receptor density of 2633 fmol/mg of protein in WP (1394 receptors/platelet; kd, 4.5 nM) and 1466 fmol/mg of protein in PM (kd, 11.3 nM). Specific binding of [3H]SQ 29,548 was inhibited by five antagonists (high/low affinity kd values in nanomolar), SQ 29,548 (WP, 5.2; PM, 7.3), SQ 28,668 (WP, 32; PM, 73), SQ 30,741 (WP, 28; PM, 50), BM 13,177 (WP, 140; PM, 4834) and BM 13,505 (WP, 5/379; PM, 11). Two agonists, U 44069 and U 46619, inhibited the binding in a biphasic manner, indicating binding to two receptor sites (approximately 20/80%) with kd values of 4/72 and 4/170 nM, respectively, in WP and 7/136 and 19/502 nM, respectively in PM. The demonstrated high affinity binding of [3H]SQ 29,548 to human platelet TP-receptors should make this radioligand a suitable and potentially useful tool in future studies of function, structure and regulation of TP-receptors.[3]
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| Cell Assay |
WST-1 assay[2]
A WST-1 assay was utilized to detect the cell viability, according to the protocol provided by the manufacturer. After transferring the cells to the plates for 24 h, the cells were treated with SQ29548 (0, 0.1, 0.5, 1.0 and 10 μM) for 2 h or treated with H2O2 (0, 0.05, 0.1, 0.2, 0.5 and 1.0 mM) for 1 h or treated with H2O2 (0, 0.1 mM) for 1 h following pretreatment with SQ29548 (0, 0.1, 0.5, 1.0 and 10 μM) for 2 h. Subsequent to the incubation for 24 h, 10 μl WST-1 solution was added to the SH-SY5Y cells. After 1 h of incubation at 37°C, a microplate reader (Bio-Rad Laboratories, Inc., Hercules, CA, USA) was used to measure the absorbance of each well at 450 nm. LDH assay[2] An LDH assay was conducted as reported previously (18). The cells were treated with H2O2 (0, 0.1 mM) for 1 h after pretreated with SQ29548 (0, 0.1, 1.0 μM) for 2 h. After 24 h of incubation, 150 μl culture medium of post-treated SH-SY5Y cells was collected and added into 96-well plates. Next, 150 μl reaction buffer containing 500 mM potassium phosphate, 7.5 mM sodium pyruvate and 1.5 mM NADH was added successively and the absorbance of each well at 340 nm was measured over 30 sec with the microplate reader. Intracellular ROS level assay[2] The intracellular ROS level was detected with DCFH-DA reagents. The cells were treated with H2O2 (0 or 0.1 mM) for 1 h following pretreatment with SQ29548 (0, 0.1, 1.0 μM) for 2 h. Treated SH-SY5Y cells were washed three times with PBS after 1-h incubation. Next, 200 μl DMEM (without sodium pyruvate, serum and antibiotics) with 10 μM DCFH-DA was added into each well. After 20 min of incubation at 37°C, DMEM (without sodium pyruvate, serum and antibiotics) was used to wash the cells for three times. The plates were then observed with an inverted fluorescence microscope at an excitation wavelength of 488 nm and an emission wavelength of 525 nm. All the data were subjected to fluorescence intensity analysis with ImageJ software, version 1.14 |
| Animal Protocol |
Either SQ 29,548 or its vehicle, 0.1 % DMSO (34869), were administered by intracerebroventricular (i.c.v.) injection 90 min and 24 h after SE (26 nmol/2 μl) (Yan et al., 2016). All animals submitted to the pilocarpine dosage protocol developed SE). Four animals died during SE before receiving diazepam or other treatments, and one vehicle-treated animal was found dead around 24 h after SE.[4]
After a 60-min stabilization period, the microvessels were constricted with serotonin (10−9–10−5 M) in the absence or presence of the selective PLA2 inhibitor quinacrine (10−6 M) or the COX inhibitor indomethacin (10−5 M) or the selective thromboxane-prostanoid (TP) receptor antagonist SQ29548 (10−6 M). Some of the microvessels were constricted with TXA-2 analog U46619 (10−9–10−6 M). One or three interventions were performed on each vessel. The order of drug administration was random.[1] |
| References |
[1]. Increased coronary arteriolar contraction to serotonin in juvenile pigs with metabolic syndrome. Mol Cell Biochem. 2019 Nov;461(1-2):57-64.
[2]. Thromboxane A2 receptor antagonist SQ29548 attenuates SH‑SY5Y neuroblastoma cell impairments induced by oxidative stress. Int J Mol Med . 2018 Jul;42(1):479-488. [3]. Characterization of [5, 6-3H] SQ 29,548 as a high affinity radioligand, binding to thromboxane A2/prostaglandin H2-receptors in human platelets. J Pharmacol Exp Ther. 1988 Jun;245(3):786-92. [4]. Neuroprotective effects of thromboxane receptor antagonist SQ 29,548 after pilocarpine-induced status epilepticus in mice. Epilepsy Res . 2020 Feb:160:106277. |
| Additional Infomation |
See also: SQ-29548 (annotation moved to).
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| Molecular Formula |
C21H29N3O4
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|---|---|
| Molecular Weight |
387.48
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| Exact Mass |
387.215
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| Elemental Analysis |
C, 65.10; H, 7.54; N, 10.84; O, 16.52
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| CAS # |
98672-91-4
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| PubChem CID |
6437074
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| Appearance |
Typically exists as White to off-white solids at room temperature
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| Density |
1.2±0.1 g/cm3
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| Index of Refraction |
1.583
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| LogP |
2.32
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
10
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| Heavy Atom Count |
28
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| Complexity |
548
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| Defined Atom Stereocenter Count |
4
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| SMILES |
C(O)(=O)CCC/C=C\C[C@@H]1[C@H](CNNC(NC2=CC=CC=C2)=O)[C@]2([H])O[C@@]1([H])CC2
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| InChi Key |
RJNDVCNWVBWHLY-YVUOLYODSA-N
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| InChi Code |
InChI=1S/C21H29N3O4/c25-20(26)11-7-2-1-6-10-16-17(19-13-12-18(16)28-19)14-22-24-21(27)23-15-8-4-3-5-9-15/h1,3-6,8-9,16-19,22H,2,7,10-14H2,(H,25,26)(H2,23,24,27)/b6-1-/t16-,17+,18+,19-/m1/s1
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| Chemical Name |
(Z)-7-[(1S,2R,3R,4R)-3-[[2-(phenylcarbamoyl)hydrazinyl]methyl]-7-oxabicyclo[2.2.1]heptan-2-yl]hept-5-enoic acid
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| Synonyms |
SQ29548; SQ 29548; (Z)-7-[(1S,2R,3R,4R)-3-[[2-(phenylcarbamoyl)hydrazinyl]methyl]-7-oxabicyclo[2.2.1]heptan-2-yl]hept-5-enoic acid; SQ-29548; SQ 29462; CHEMBL365308; DTXSID50873403; SQ-29548
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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 |
| 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) |
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
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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 | 2.5808 mL | 12.9039 mL | 25.8078 mL | |
| 5 mM | 0.5162 mL | 2.5808 mL | 5.1616 mL | |
| 10 mM | 0.2581 mL | 1.2904 mL | 2.5808 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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