| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Other Sizes |
| Targets |
GPX4[1]; Ferroptosis[1]
Glutathione peroxidase 4 (GPX4) |
|---|---|
| ln Vitro |
Cell viability is inhibited by JKE-1716 (0~100 nM; 72 hours; LOX-IMVI cells)[1]. JKE-1716 (LOX-IMVI cells) suppresses α-GPX4 expression [1].
In vitro, JKE-1716 inhibits cell viability in LOX-IMVI melanoma cells in a concentration-dependent manner, an effect that can be completely blocked by the ferroptosis inhibitor ferrostatin-1, confirming the mechanism of action via ferroptosis induction. The compound is active against a panel of additional cancer cell lines. JKE-1716 suppresses alpha-GPX4 expression and induces ferroptosis through covalent GPX4 inhibition. Cell viability is inhibited by JKE-1716 at concentrations of 0-100 nM over 72 hours in LOX-IMVI cells. The compound is a potent and specific nitrolic acid-containing GPX4 inhibitor. |
| ln Vivo |
No specific in vivo data for JKE-1716; however, as a GPX4 inhibitor and ferroptosis inducer, it has potential for in vivo efficacy in xenograft models of melanoma and other cancers. GPX4 inhibition leads to accumulation of lipid peroxides, ultimately causing iron-dependent cell death (ferroptosis). In vivo studies would involve administration to tumor-bearing mice to assess tumor growth inhibition, survival, and modulation of GPX4 activity and lipid peroxidation (MDA, 4-HNE levels) in tumor tissues. The compound is likely to have moderate oral bioavailability based on its molecular properties.
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| Enzyme Assay |
Recombinant human GPX4 is expressed in E. coli or insect cells and purified. The GPX4 activity assay measures the reduction of lipid hydroperoxides or small molecule peroxides using glutathione (GSH) as a reducing agent. A typical coupled assay uses cumene hydroperoxide or phosphatidylcholine hydroperoxide as substrate and monitors NADPH oxidation (decrease in absorbance at 340 nm) in the presence of glutathione reductase. Alternatively, a peroxidase activity assay using tert-butyl hydroperoxide and a fluorogenic probe (Amplex Red) can be used. JKE-1716 is pre-incubated with GPX4 (10-100 nM) for 10-30 minutes to allow for covalent binding. IC50 values are determined from dose-response curves. Covalent modification can be confirmed by LC-MS/MS analysis of the tryptic digest or by activity-based protein profiling (ABPP) using a biotinylated probe. The nitrolic acid warhead forms a covalent adduct with the active site selenocysteine (Sec) residue of GPX4.
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| Cell Assay |
Cell Viability Assay[1]
Cell Types: LOX-IMVI cells Tested Concentrations: 0~100 nΜ Incubation Duration: 72 hrs (hours) Experimental Results: Inhibited cell viability. Human cancer cell lines (e.g., LOX-IMVI melanoma, HT1080 fibrosarcoma, PANC-1 pancreatic cancer) are cultured in RPMI-1640 or DMEM with 10% FBS and antibiotics. Cells are seeded in 96-well plates and treated with JKE-1716 (0.1-1000 nM) for 48-72 hours. Cell viability is assessed by MTT, CCK-8, or CellTiter-Glo assays. For ferroptosis specificity studies, cells are co-treated with ferroptosis inhibitors (ferrostatin-1 1-10 uM, liproxstatin-1 1-10 uM) or with necrosis (necrostatin-1), apoptosis (Z-VAD-FMK), and autophagy (3-methyladenine) inhibitors. Ferroptosis is confirmed by rescue of cell death only with ferroptosis inhibitors. Cellular GPX4 activity is measured in cell lysates using a GPX4 activity assay kit. Lipid peroxidation is assessed by C11-BODIPY581/591 fluorescence staining (oxidation shifts emission from red to green) followed by flow cytometry or fluorescence microscopy. Cellular malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE) levels are measured by colorimetric or LC-MS/MS methods. Total glutathione (GSH) and oxidized glutathione (GSSG) levels are measured using enzymatic recycling assays. Cell death is quantified by Annexin V/PI staining and flow cytometry. Caspase-3/7 activity is measured by luminescence assays to rule out apoptosis. For alpha-GPX4 expression, Western blot analysis is performed using anti-GPX4 antibody. |
| Animal Protocol |
No published in vivo animal study for JKE-1716. Based on its GPX4 inhibition and ferroptosis-inducing activity, a typical protocol would involve establishing subcutaneous tumor xenografts in immunodeficient mice (e.g., nude mice or NSG mice) using LOX-IMVI, HT1080, or other sensitive cancer cell lines. When tumors reach 100-200 mm3, mice are randomized and treated with JKE-1716 formulated in a suitable vehicle (e.g., 10% DMSO + 40% PEG300 + 5% Tween 80 + 45% saline, or 10% DMSO + 90% corn oil). Dosing is once daily by intraperitoneal injection (IP) or oral gavage at doses ranging from 1-50 mg/kg for 2-4 weeks. Endpoints: tumor volume measured by calipers, tumor weight at necropsy, body weight, and survival. Plasma and tumor tissue are collected for pharmacokinetic analysis and pharmacodynamic evaluation: GPX4 activity in tumor lysates (using activity assay), lipid peroxidation (MDA, 4-HNE, C11-BODIPY), GSH/GSSG ratio, and ferritin/transferrin receptor levels (by Western blot). Histological analysis of tumors (H&E, Prussian blue for iron, TUNEL for cell death, Ki-67 for proliferation) and normal tissues (liver, kidney) for toxicity assessment. Ferroptosis can be confirmed by co-administration of ferrostatin-1 (10 mg/kg IP) which should block the antitumor effect.
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| ADME/Pharmacokinetics |
No specific pharmacokinetic data for JKE-1716. Based on its molecular properties (MW 451.30, LogP ~3-4 from structure), it is likely to have moderate to high lipophilicity, leading to high plasma protein binding and extensive tissue distribution. The compound contains a nitrolic acid warhead that is reactive and may undergo rapid metabolism via reduction or conjugation. Oral bioavailability may be moderate due to first-pass metabolism. For in vivo studies, intraperitoneal (IP) administration is often preferred for reactive covalent inhibitors to achieve consistent exposure. Plasma half-life, Cmax, AUC, clearance, volume of distribution, and oral bioavailability have not been publicly reported. Solubility: soluble in DMSO. Stock solutions can be stored at -20degC. In vivo formulation: 10% DMSO + 40% PEG300 + 5% Tween80 + 45% saline is a common vehicle for similar compounds.
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| Toxicity/Toxicokinetics |
No specific toxicity data for JKE-1716 in published sources. As a GPX4 inhibitor and ferroptosis inducer, the primary expected on-target toxicity is induction of ferroptosis in normal tissues, particularly in organs with high metabolic activity and iron content, such as liver, kidney, and pancreas. GPX4 is essential for survival of certain cell types, and systemic GPX4 knockout in mice is lethal. Therefore, toxicity is expected at effective doses, and the therapeutic window may be narrow. In cell-based studies, JKE-1716 reduces viability of multiple cancer cell lines, and toxicity is rescued by ferrostatin-1. In vivo toxicity studies would assess maximum tolerated dose (MTD), body weight loss, organ toxicity (liver enzymes ALT/AST, kidney function BUN/creatinine, pancreatic enzymes), and histopathological changes in normal tissues. No data are publicly available. For research use only, handle with extreme caution.
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| References | |
| Additional Infomation |
JKE-1716 (CAS: 2421118-05-8) is a potent and selective GPX4 inhibitor that induces ferroptosis, an iron-dependent form of regulated cell death characterized by lipid peroxidation. It is a derivative of ML-210 and contains a nitrolic acid scaffold that covalently targets the active site selenocysteine of GPX4. JKE-1716 reduces viability of LOX-IMVI cancer cells and a panel of additional cancer cell lines, an effect blocked by ferrostatin-1. Molecular formula: C20H20Cl2N4O4, molecular weight: 451.30. Appearance: solid. Solubility: soluble in DMSO. Storage: powder at -20degC for 3 years; in solvent at -80degC for 6 months. It is a research tool for studying ferroptosis, iron metabolism, and cancer biology. Not approved for clinical use. References: Eaton JK, et al. Selective covalent targeting of GPX4 using masked nitrile-oxide electrophiles. Nat Chem Biol. 2020;16(5):497-506. For research use only.
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| Molecular Formula |
C20H20CL2N4O4
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|---|---|
| Molecular Weight |
451.30
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| Exact Mass |
450.086
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| CAS # |
2421118-05-8
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| PubChem CID |
145865953
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| Appearance |
Off-white to light yellow solid powder
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| Density |
1.4±0.1 g/cm3
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| Boiling Point |
641.1±65.0 °C at 760 mmHg
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| Flash Point |
341.5±34.3 °C
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| Vapour Pressure |
0.0±2.0 mmHg at 25°C
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| Index of Refraction |
1.651
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| LogP |
3.84
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
30
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| Complexity |
599
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1CN(CCN1C(C2=CC=C(C=C2)Cl)C3=CC=C(C=C3)Cl)C(=O)C/C(=N/O)/[N+](=O)[O-]
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| InChi Key |
HZFRTZXJNACSIE-NKFKGCMQSA-N
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| InChi Code |
InChI=1S/C20H20Cl2N4O4/c21-16-5-1-14(2-6-16)20(15-3-7-17(22)8-4-15)25-11-9-24(10-12-25)19(27)13-18(23-28)26(29)30/h1-8,20,28H,9-13H2/b23-18-
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| Chemical Name |
(3Z)-1-[4-[bis(4-chlorophenyl)methyl]piperazin-1-yl]-3-hydroxyimino-3-nitropropan-1-one
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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 | 2.2158 mL | 11.0791 mL | 22.1582 mL | |
| 5 mM | 0.4432 mL | 2.2158 mL | 4.4316 mL | |
| 10 mM | 0.2216 mL | 1.1079 mL | 2.2158 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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