| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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Purity: ≥98%
CYM-5541 (also known as ML249) is a potent, selective, full, and allosteric agonist of the S1P3 (Sphingosine 1-phosphate) receptor with EC50 in the range of 72 to 132 nM. S1P is a lysophospholipid signaling molecule that activates G protein-coupled receptors for S1P, specifically S1P(1) through S1P(5), to regulate key biological processes, such as lymphocyte trafficking and vascular development. Phe263 is an important gate-keeper residue for the affinity and efficacy of an allosteric site that was identified using CYM-5541. This ligand is nonpolar, and the unique allosteric hydrophobic pocket allows CYM-5541's S1P(3) selectivity within the remarkably similar S1P receptor family.
| Targets |
S1P3 ( EC50: between 72 and 132 nM )
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| ln Vivo |
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| Enzyme Assay |
For four hours, WT or mutant S1P3-expressing Jump-In TI CHO-K cells are serum-starved. Following this, they are incubated for 30 minutes at 4 °C in the binding buffer, which contains 20 mM Tris-HCl (pH 7.5), 100 mM NaCl, 15 mM NaF, 0.5 mM EDTA, 1 mM Na2VO4, 0.5% fatty acid-free bovine serum albumin, and a protease inhibitor mixture containing 0.1 nM [33P]S1P and increasing concentrations of S1P, SPM-242, or CYM-5541. Cold binding buffer is used to wash the cells three times. Liquid scintillation counting is the method used to measure cell-bound radioactivity after the cells have been lysed with 0.5% SDS. In order to reference the level of [33P]S1P bound to each cell line (WT or mutant) in the absence of a competing ligand as 100% for its own cell line, the raw data is normalized[1].
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| Cell Assay |
The pcDNA3.1 plasmid encoding the triple HA-tagged N-terminal S1P3 fusion protein was acquired from cDNA.org. Overlapping PCR mutagenesis was used to create S1P3 receptor mutants, and sequences were checked before use. The Gateway cloning vectors (pDONR 221, pJTI R4 DEST, and pJTI R4 Int), enzymes (BP clonase II and LR clonase), and Jump-InTM TI™ CHO-K parental cells were acquired from Invitrogen Corp. The BP recombination reaction was first used to clone triple HA-tagged WT and mutant S1P3 into entry clones. In order to create a pJTI R4 EXP retargeting expression vector, they were retargeted into a suitable pJTI R4 DEST vector. For retargeting, both the pJTI R4 EXP S1P1 and the pJTI R4 Int vector were transfected into Jump-In TI CHO-K cells. Retargeted Jump-In TI cells were selected using blasticidin at 10 μg/mL for four weeks after the cells had expanded.
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| Animal Protocol |
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| References |
| Molecular Formula |
C19H28N2O2
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| Molecular Weight |
316.45
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| Exact Mass |
316.215
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| Elemental Analysis |
C, 72.12; H, 8.92; N, 8.85; O, 10.11
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| CAS # |
945128-26-7
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| Related CAS # |
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| PubChem CID |
17253208
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| Appearance |
White to off-white solid powder
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| Density |
1.1±0.1 g/cm3
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| Boiling Point |
503.1±38.0 °C at 760 mmHg
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| Flash Point |
258.1±26.8 °C
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| Vapour Pressure |
0.0±1.3 mmHg at 25°C
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| Index of Refraction |
1.559
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| LogP |
3.47
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
23
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| Complexity |
394
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O=C(N(C1CCCCC1)C1CCCCC1)C1C=C(C2CC2)ON=1
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| InChi Key |
NDKGACIWVAOUQH-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C19H28N2O2/c22-19(17-13-18(23-20-17)14-11-12-14)21(15-7-3-1-4-8-15)16-9-5-2-6-10-16/h13-16H,1-12H2
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| Chemical Name |
N,N-dicyclohexyl-5-cyclopropyl-1,2-oxazole-3-carboxamide
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| Synonyms |
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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 |
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| 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) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (7.90 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 25.0 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. Solubility in Formulation 2: ≥ 2.5 mg/mL (7.90 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 25.0 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (7.90 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.1601 mL | 15.8003 mL | 31.6006 mL | |
| 5 mM | 0.6320 mL | 3.1601 mL | 6.3201 mL | |
| 10 mM | 0.3160 mL | 1.5800 mL | 3.1601 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.
Ligand-induced ERK phosphorylation in WT S1P3Jump-In stable cell lines (Mean ± SEM; n=3), revealing CYM-5541 is a full agonist of S1P3 |
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Radioligand binding and competition. |
Receptor mutagenesis and ligand-induced ERK phosphorylation. |
SPM-242, an S1P3-selective antagonist, is a bitopic antagonist. |
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Visualization of the receptor binding pocket by homology modeling and docking. Three dimensional plot of S1P3binding to (A) S1P, (B) CYM-5541, and (C and D) both |
Visualization of the orthosteric, allosteric, and bitopic ligand interactions based upon the biochemistry, mutagenesis, homology modeling, and molecular dynamic simulations.CYM-5541 (green) and SPM-242 (cyan) overlap in the S1P3binding pocket. |
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