| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| Other Sizes |
| Targets |
NEDD8 activating enzyme (NAE)[1]
NEDD8-activating enzyme (NAE). ZM223 hydrochloride is a potent, non-covalent, and orally active inhibitor of NAE, the E1 enzyme that initiates the neddylation cascade. |
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| ln Vitro |
With IC50s of 100 and 122 nM, respectively, ZM223 hydrochloride (0.1–1 μM; 4 hours) inhibits HCT-116 and U-2OS cancer cells[1]. The administration of ZM223 hydrochloride (0.1–1 μM) for four hours results in a dose-dependent reduction in NEDD8 levels and an accumulation of UBC12 protein, which suggests a decrease in the NEDD8-UBC12 complex that follows[1].
In vitro, ZM223 hydrochloride (0.1-1 microM; 4 hours) potently inhibits the proliferation of HCT-116 and U-2OS cancer cells with IC50 values of 100 nM and 122 nM, respectively. It causes a dose-dependent decrease in the level of NEDD8 and an accumulation of the UBC12 protein, indicating a decrease in the NEDD8-UBC12 complex and inhibition of the neddylation pathway. |
| ln Vivo |
In vivo, ZM223 hydrochloride is orally active and has excellent anticancer activity. It has been evaluated in mouse xenograft models of cancer, where it likely inhibits tumor growth via NAE inhibition. Specific in vivo data (e.g., tumor growth inhibition percentages) are not detailed in the available search results, but the compound is described as having excellent in vivo efficacy.
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| Enzyme Assay |
For non-cellular assays, the activity of NAE can be measured by incubating the purified enzyme with NEDD8 and ATP. The formation of the NEDD8-NAE thioester intermediate can be detected by gel electrophoresis under non-reducing conditions followed by Western blot. ZM223 hydrochloride is added at varying concentrations (1-1000 nM). The inhibition of the thioester intermediate formation can be quantified, and an IC50 value can be determined.
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| Cell Assay |
Cell Viability Assay[1]
Cell Types: HCT116 colon cancer cells and U-2OS osteosarcoma cells Tested Concentrations: 0.1 μM, 1 μM Incubation Duration: 4 hrs (hours) Experimental Results: Inhibited both HCT-116 and U-2OS cancer cells. Western Blot Analysis[1] Cell Types: HCT116 colon cancer cells Tested Concentrations: 0.1 μM, 1 μM Incubation Duration: 4 hrs (hours) Experimental Results: Caused a decrease in the level of NEDD8 and an increase in the downstream UBC12 protein. For cell-based assays, cancer cells (e.g., HCT-116 or U-2OS) are treated with ZM223 hydrochloride (0.01-10 uM) for 4-24 hours. The levels of neddylated proteins (e.g., NEDD8-cullins) and the accumulation of UBC12 are assessed by Western blot. Cell proliferation and viability are measured using MTT or CellTiter-Glo assays. Apoptosis is assessed by measuring caspase-3/7 activity or by flow cytometry using Annexin V staining. |
| Animal Protocol |
For animal studies, ZM223 hydrochloride is administered orally to mice bearing subcutaneous human tumor xenografts (e.g., HCT-116 colon cancer xenografts). Doses and dosing regimens would need to be optimized, but typical doses for NAE inhibitors are in the range of 10-100 mg/kg given daily or on a schedule (e.g., 3 days on, 1 day off). Tumor volume is measured by calipers, and tumor growth inhibition (TGI) is calculated. At the end of the study, tumors are harvested, and the levels of neddylated proteins and UBC12 accumulation are analyzed by Western blot.
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| ADME/Pharmacokinetics |
Pharmacokinetic data for ZM223 hydrochloride are not detailed in the available literature. It is described as an orally active compound, meaning it is absorbed from the gastrointestinal tract and has sufficient bioavailability to exert its effects. The compound has a molecular formula of C23H18ClF3N4O2S2 and a molecular weight of 538.99 g/mol. It is typically formulated in a vehicle such as 10% DMSO, 40% PEG300, 5% Tween-80, and 45% saline for in vivo administration.
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| Toxicity/Toxicokinetics |
Specific toxicological data for ZM223 hydrochloride are not detailed in the available literature. The compound is intended for research use only and is not approved for human use. Material safety data sheets may be available for the compound, and standard safety precautions should be observed, including the use of gloves and eye protection and working in a fume hood. The compound may be irritating to the eyes.
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| References | |
| Additional Infomation |
ZM223 hydrochloride is a second-generation, non-covalent NAE inhibitor. It differs from first-generation NAE inhibitors such as Pevonedistat (MLN4924), which are covalent inhibitors. Non-covalent inhibitors may have a different safety and resistance profile. ZM223 hydrochloride is a valuable research tool for studying the neddylation pathway and its role in cancer biology. It is not an FDA-approved drug and is intended for research use only.
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| Molecular Formula |
C23H18CLF3N4O2S2
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|---|---|
| Molecular Weight |
538.992831707001
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| Exact Mass |
538.051
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| CAS # |
2438679-27-5
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| Related CAS # |
ZM223;2031177-48-5
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| PubChem CID |
139035004
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
35
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| Complexity |
713
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1=CC(=CC=C1C(=O)NC2=NC3=C(S2)C=C(C=C3)NC(=O)CSC4=CC=C(C=C4)N)C(F)(F)F.Cl
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| InChi Key |
DXCZCDSLNIYJQF-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C23H17F3N4O2S2.ClH/c24-23(25,26)14-3-1-13(2-4-14)21(32)30-22-29-18-10-7-16(11-19(18)34-22)28-20(31)12-33-17-8-5-15(27)6-9-17;/h1-11H,12,27H2,(H,28,31)(H,29,30,32);1H
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| Chemical Name |
N-[6-[[2-(4-aminophenyl)sulfanylacetyl]amino]-1,3-benzothiazol-2-yl]-4-(trifluoromethyl)benzamide;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: 150 mg/mL (278.30 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 7.5 mg/mL (13.91 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 75.0 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: 7.5 mg/mL (13.91 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 75.0 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.8553 mL | 9.2766 mL | 18.5532 mL | |
| 5 mM | 0.3711 mL | 1.8553 mL | 3.7106 mL | |
| 10 mM | 0.1855 mL | 0.9277 mL | 1.8553 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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