| Size | Price | Stock | Qty |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg | |||
| 500mg | |||
| Other Sizes |
| Targets |
CCR8 ( IC50 = 1.8 μM ); 5-HT1A ( IC50 = 5.4 μM ); 5-HT2B ( IC50 = 4.4 μM ); 5-HT2C ( IC50 = 34.8 μM ); 5-HT5A ( IC50 = 16 μM ); 5-HT6 ( IC50 = 5.9 μM ); α2A ( IC50 < 20 μM )
CC chemokine receptor CCR8 (also known as C-C chemokine receptor type 8). ZK-756326 acts as a selective and nonpeptide agonist of CCR8, inhibiting the binding of the natural ligand I-309 (CCL1) and activating receptor-mediated signaling pathways. The compound shows high specificity for CCR8 with >28-fold selectivity over 26 other GPCRs tested. |
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| ln Vitro |
ZK 756326 prevents the CCR8 ligand I-309 (CCL1) from binding, with an IC50 value of 1.8 μM. ZK 756326 is a full agonist of CCR8, which cross-desensitizes the receptor's response to CCL1 and dose-responsively increases intracellular calcium. Extracellular acidification is stimulated in human CCR8-expressing cells by ZK 756326. To find out if ZK 756326's interaction is unique to CCR8, binding competition tests are run on a number of other G-protein-coupled receptors. In these tests, the inhibitory effect of ZK 756326 at 50 μM on radiolabeled ligand binding is examined. ZK 756326 exhibits >28 fold specificity for CCR8 at this concentration when compared to 26 other GPCRs, all of which have IC50 values greater than 50 μM. ZK 756326 exhibits reduced selectivity when tested against the adrenergic receptor α2A, the serotonergic receptors 5-HT1A, 5-HT2B, 5-HT2C, 5-HT5A, and 5-HT6, with IC50 values of 5.4, 4,4, 34.8, 16, 5.9, and <20 μM (at 20 μM 65% inhibition), respectively. Since the compound does not exhibit any agonist activity in a GTPγS binding assay when tested at concentrations up to 10 μM on the representative receptor 5-HT1A, it is unlikely to be an agonist on these biogenic amine receptors[1].
ZK-756326 inhibits the binding of the CCR8 ligand I-309 (CCL1) with an IC50 of 1.8 μM. As a full agonist of CCR8, it dose-responsively elicits an increase in intracellular calcium and cross-desensitizes the receptor's response to CCL1. The compound stimulates extracellular acidification in cells expressing human CCR8, indicating metabolic activation. It shows minimal activity against other GPCRs, with IC50 values >50 μM for most receptors tested. |
| ln Vivo |
In vivo activity data for ZK-756326 are limited in the available literature. As a CCR8 agonist, it is expected to modulate immune cell trafficking and chemotaxis in animal models. The compound has been shown to induce chemotaxis in murine CCR8-expressing cells, suggesting potential in vivo activity in immune response studies. Further in vivo studies are needed to fully characterize its pharmacological effects and therapeutic potential in disease models.
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| Enzyme Assay |
The in vitro receptor binding assay for ZK-756326 typically involves competition binding experiments using radiolabeled CCR8 ligands such as I-309 (CCL1). Membranes from cells expressing human CCR8 are incubated with the radiolabeled ligand and varying concentrations of the test compound. The IC50 value is determined by measuring the displacement of the radiolabeled ligand. Binding specificity is assessed by testing the compound against a panel of other G-protein-coupled receptors at concentrations up to 50 μM.
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| Cell Assay |
U87 MG cells that express CCR8 are plated at a density of 10,000 cells per well on black 96-well plates coated with poly-D-lysine and cultured for an entire night. After that, cells are loaded with Calcium 3, a Ca2+-sensitive non-wash fluorescence dye, and allowed to sit in Hanks' balanced salts solution (which contains 2.5 mM probenecid, 3.2 mM CaCl22, 1% (v/v) fetal bovine serum, and 20 mM HEPES) for 60 minutes at 37°C. Fluorometric Imaging Plate Reader (FLIPR 3) is used to measure intracellular free-Ca2+ concentration changes as soon as the agonist is added at room temperature. In cross-desensitization experiments, the agonist (CCL1 at 30 nM or ZK 756326 at 3 μM) is added once, and then 100 nM CCL1 is added right away after[1].
The in vitro cellular assay for ZK-756326 typically involves the use of cells expressing human CCR8, such as transfected cell lines. The compound is tested for its ability to stimulate intracellular calcium mobilization using fluorescent calcium indicators (e.g., Fluo-4). The agonist activity is confirmed by measuring the dose-dependent increase in intracellular calcium and by assessing cross-desensitization of the receptor response to CCL1. Extracellular acidification rates can also be measured as a readout of metabolic activity. |
| Animal Protocol |
In vivo animal studies for ZK-756326 would typically involve the use of mouse models to assess the compound's effects on immune cell trafficking and chemotaxis. Mice would be administered the compound via appropriate routes (e.g., intraperitoneal or subcutaneous injection), and the migration of CCR8-expressing cells to sites of inflammation or infection would be monitored. Endpoints include analysis of immune cell populations in tissues and assessment of functional responses such as chemotaxis inhibition.
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| ADME/Pharmacokinetics |
Detailed pharmacokinetic properties of ZK-756326 are not extensively reported in the available literature. As a small molecule with a molecular weight of 429.38 g/mol, it is expected to have reasonable bioavailability and tissue distribution. The compound shows good solubility in water (50 mg/mL) and DMSO (27.5 mg/mL), indicating favorable physicochemical properties for in vitro and in vivo studies. Further PK studies are needed to determine parameters such as half-life and clearance.
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| Toxicity/Toxicokinetics |
Specific toxicology data for ZK-756326 are not available in the public domain. The compound is intended for research use only and should be handled with appropriate laboratory safety precautions. Standard safety assessments would be required before any clinical application. Researchers should consult the safety data sheet (SDS) for detailed handling and disposal information.
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| References | |
| Additional Infomation |
See also: ZK 756326 (Notes have been moved to).
ZK-756326 is also known as ZK756326 and is commercially available as a research chemical. It is a full agonist of CCR8 that works through Gαi-mediated signaling pathways, as its effects can be blocked by pertussis toxin treatment. The compound is useful for studying CCR8 biology where the use of the natural protein agonist (CCL1) is impractical. It has not yet entered clinical trials and is strictly for preclinical research purposes. |
| Molecular Formula |
C21H30CL2N2O3
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|---|---|
| Molecular Weight |
429.380504131317
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| Exact Mass |
428.16
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| Elemental Analysis |
C, 58.74; H, 7.04; Cl, 16.51; N, 6.52; O, 11.18
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| CAS # |
1780259-94-0
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| Related CAS # |
ZK 756326; 874911-96-3
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| PubChem CID |
56972201
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
9
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| Heavy Atom Count |
28
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| Complexity |
368
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
MPACCEKWFGWZHS-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C21H28N2O3.2ClH/c24-14-16-25-15-13-22-9-11-23(12-10-22)18-19-5-4-8-21(17-19)26-20-6-2-1-3-7-20;;/h1-8,17,24H,9-16,18H2;2*1H
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| Chemical Name |
2-[2-[4-[(3-phenoxyphenyl)methyl]piperazin-1-yl]ethoxy]ethanol;dihydrochloride
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| Synonyms |
ZK756326; ZK-756326; ZK 756326; ZK-756326 dihydrochloride
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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: ~27.5 mg/mL (~64.1 mM)
H2O: ~50 mg/mL (~116.5 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.75 mg/mL (6.40 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 27.5 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. Solubility in Formulation 2: ≥ 2.75 mg/mL (6.40 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 27.5 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. View More
Solubility in Formulation 3: 50 mg/mL (116.45 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3289 mL | 11.6447 mL | 23.2894 mL | |
| 5 mM | 0.4658 mL | 2.3289 mL | 4.6579 mL | |
| 10 mM | 0.2329 mL | 1.1645 mL | 2.3289 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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