| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
IC50: 79 nM (human uPA), 1580 nM (human tPA), 1330 nM (human plasmin)[1].
Urokinase plasminogen activator (uPA), with IC50s of 79 nM for human uPA. It also inhibits tPA (1580 nM) and plasmin (1330 nM) at higher concentrations, showing selectivity for uPA. |
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| ln Vitro |
ZK824859 has lost selectivity and is five times less potent in mice than in humans, with IC50 values of 79 nM, 1580 nM, and 1330 nM for uPA, tPA, and plasmin, respectively, of 410 nM, 910 nM, and 1600 nM[1].
In vitro, ZK824859 hydrochloride is a selective inhibitor of uPA, with an IC50 of 79 nM for the human enzyme. It has a weaker effect on tPA (IC50 = 1580 nM) and plasmin (IC50 = 1330 nM), demonstrating good selectivity. The compound is 5-fold less potent and loses selectivity in mouse enzymes compared to human, with IC50 values of 410 nM, 910 nM, and 1600 nM for mouse uPA, tPA, and plasmin, respectively. |
| ln Vivo |
In a chronic mouse EAE model, ZK824859 (50, 25 and 10 mg/kg; bid; 25 days) is employed. ZK 824859 totally stops illness from starting. However, clinical ratings are unaffected by two lower doses (25 and 10 mg/kg)[1].
In a chronic mouse model of experimental autoimmune encephalomyelitis (EAE) of multiple sclerosis, ZK824859 (50 mg/kg, b.i.d.) completely prevented the development of the disease, while lower doses (25 and 10 mg/kg) had no effect. This demonstrates the in vivo efficacy of uPA inhibition in an animal model of neuroinflammation. It also shows that ZK824859 is orally active and has potential for treating MS. |
| Enzyme Assay |
For non-cellular assays, the enzymatic activity of human uPA is measured using a fluorogenic tripeptide substrate, such as Z-Gly-Gly-Arg-AMC. The enzyme and substrate are incubated in an assay buffer (e.g., 50 mM Tris-HCl, pH 7.4, 150 mM NaCl). ZK824859 is added at varying concentrations (1-1000 nM). The release of AMC is measured (excitation 380 nm, emission 460 nm) to calculate the inhibition of uPA activity. IC50 values for tPA and plasmin are determined similarly.
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| Cell Assay |
For cell-based assays, human cancer cell lines that express uPA and invade through Matrigel (e.g., MDA-MB-231 breast cancer cells) are treated with ZK824859 (10-1000 nM). The cells are seeded onto Matrigel-coated transwell inserts and allowed to invade towards a chemoattractant. The number of invaded cells is quantified after 24-48 hours. The level of uPA activity in conditioned media can also be measured using the fluorogenic substrate.
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| Animal Protocol |
Animal/Disease Models: Female SJL mice with chronic mouse EAE model[1]
Doses: 50, 25 and 10 mg/kg Route of Administration: Bid; 25 days Experimental Results: Prevented the development of disease at 50 mg/kg completely. However, 25 and 10 mg/kg had no effect on clinical scores. For animal studies, ZK824859 can be administered orally to the mouse EAE model (chronic, relapsing-remitting). Doses are typically given twice daily (b.i.d.) at 10-50 mg/kg. Clinical scores are monitored daily to assess disease severity. The compound can be formulated as a suspension in a vehicle such as 0.5% methylcellulose. At the end of the study, spinal cord and brain tissues are collected for histopathological analysis of demyelination and immune cell infiltration. uPA activity in the tissue is measured. |
| ADME/Pharmacokinetics |
ZK824859 is orally bioavailable, with an oral bioavailability of 55% in rats and a half-life of 2.8 hours. It is a direct-acting inhibitor, meaning it does not require metabolic activation to exert its pharmacological effect, unlike prodrug-based uPA inhibitors. The compound has a molecular weight of 464.89 g/mol and a molecular formula of C23H22F2N4O4·HCl.
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| Toxicity/Toxicokinetics |
Toxicological data for ZK824859 are limited. According to safety data sheets, the compound is toxic and may cause serious damage to eyes. In the chronic EAE model, the 50 mg/kg dose was well-tolerated, with no reports of overt toxicity. Standard safety precautions, including the use of gloves and eye protection, should be followed.
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| References | |
| Additional Infomation |
ZK824859 is an orally active uPA inhibitor that has shown promising efficacy in the EAE model of MS. The hydrochloride salt form has enhanced water solubility and stability compared to the free base. The compound is a research tool and is not an approved drug. It is a valuable tool for studying the role of the uPA system in multiple sclerosis, cancer metastasis, and other inflammatory diseases.
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| Molecular Formula |
C23H23CLF2N2O4
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|---|---|
| Molecular Weight |
464.889532327652
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| Exact Mass |
464.131
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| CAS # |
2436760-76-6
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| Related CAS # |
ZK824859;2271122-53-1
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| PubChem CID |
137333452
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| Appearance |
Light yellow to yellow solid powder
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
32
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| Complexity |
584
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| Defined Atom Stereocenter Count |
1
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| SMILES |
CC[C@H](C(=O)O)OC1=C(C=C(C(=N1)OC2=CC(=CC(=C2)C3=CC=CC(=C3)CN)C)F)F.Cl
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| InChi Key |
XACCKEPNGBUXQW-VEIFNGETSA-N
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| InChi Code |
InChI=1S/C23H22F2N2O4.ClH/c1-3-20(23(28)29)31-22-19(25)11-18(24)21(27-22)30-17-8-13(2)7-16(10-17)15-6-4-5-14(9-15)12-26;/h4-11,20H,3,12,26H2,1-2H3,(H,28,29);1H/t20-;/m1./s1
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| Chemical Name |
(2R)-2-[6-[3-[3-(aminomethyl)phenyl]-5-methylphenoxy]-3,5-difluoropyridin-2-yl]oxybutanoic acid;hydrochloride
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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: ≥ 125 mg/mL (268.88 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.47 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 20.8 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: ≥ 2.08 mg/mL (4.47 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 20.8 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: ≥ 2.08 mg/mL (4.47 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 | 2.1510 mL | 10.7552 mL | 21.5105 mL | |
| 5 mM | 0.4302 mL | 2.1510 mL | 4.3021 mL | |
| 10 mM | 0.2151 mL | 1.0755 mL | 2.1510 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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