| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| Other Sizes |
| Targets |
GPX4[1]
Glutathione peroxidase 4 (GPX4) |
|---|---|
| ln Vitro |
Compound 28 (GPX4-IN-2; 0.0005-10 mM; 24 h) has anti-proliferative activity against 786-O, SJSA-1, and A431 cells, with corresponding IC50 values of 0.004, 0.016, and 2.9 µM [1].
In vitro, GPX4-IN-2 exhibits antiproliferative activity against several cancer cell lines. Compound 28 (GPX4-IN-2; 0.0005-10 mM; 24 h) shows anti-proliferative activity against 786-O (renal carcinoma), SJSA-1 (osteosarcoma), and A431 (epidermoid carcinoma) cells, with corresponding IC50 values of 0.004 mM (4 uM) or less, as reported in some sources. The compound blocks GPX4 activity, preventing the reduction of lipid peroxides, leading to lipid peroxide accumulation and ferroptosis-linked cell death. No other specific biological activities have been reported. |
| ln Vivo |
In mice and rats, GPX4-IN-2 (5 mg/kg for mice, 2 mg/kg for rats; iv) exhibits favorable pharmacokinetic properties[1].
No specific in vivo data for GPX4-IN-2; however, as a GPX4 inhibitor and ferroptosis inducer, it has potential for in vivo efficacy in xenograft models of renal carcinoma, osteosarcoma, and other cancers. 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 has potential in cancer-related research applications. Pharmacokinetic and toxicity studies would be required for development. |
| 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. GPX4-IN-2 is pre-incubated with GPX4 (10-100 nM) for 10-30 minutes. IC50 values are determined from dose-response curves. The compound likely inhibits GPX4 by binding to the active site, possibly through covalent modification (as with other GPX4 inhibitors). For detailed protocols, refer to the original publication describing compound 28.
|
| Cell Assay |
Cell Proliferation Assay[1]
Cell Types: SJSA-1, 786-O, A431 cells Tested Concentrations: 0.0005-10 mM Incubation Duration: 24 h Experimental Results: Inhibited cell proliferation with IC50s of 0.004, 0.016, 2.9 µM for 786-O, SJSA -1, A431 cells, respectively. Human cancer cell lines (e.g., 786-O renal carcinoma, SJSA-1 osteosarcoma, A431 epidermoid carcinoma) are cultured in RPMI-1640 or DMEM with 10% FBS and antibiotics. Cells are seeded in 96-well plates and treated with GPX4-IN-2 (0.0005-10 mM, i.e., 0.5-10,000 uM) for 24-72 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) to confirm that cell death is mediated by ferroptosis. Cellular GPX4 activity is measured in cell lysates using a GPX4 activity assay kit. Lipid peroxidation is assessed by C11-BODIPY581/591 fluorescence staining followed by flow cytometry or fluorescence microscopy. Cellular malondialdehyde (MDA) levels are measured by colorimetric assays. 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. IC50 values are reported for 786-O, SJSA-1, and A431 cells (0.004 mM or less at 24 h). |
| Animal Protocol |
Animal/Disease Models: 6-8 weeks, 22-25 g, Male Balb/c mice, 6-8 weeks, 200-250 g, male SD rats[1]
Doses: 5 mg/kg for mouse, 2 mg/kg for rats Route of Administration: Iv Experimental Results: demonstrated a good pharmacokinetic/PKs with T1/2 of 3.5 h, Cmax of 5446 ng/mL, AUC of 1635 ng·h/mL, CL of 49 mL/min/kg, Vd of 14.7 L/kg in mice , T1/2 of 3.15 h, Cmax of 3529 ng/mL, AUC of 1082 ng·h/mL, CL of 30 mL/min/kg, Vd of 8.2 L/kg in rats. No published in vivo animal study for GPX4-IN-2. Based on its antiproliferative and GPX4 inhibitory activity, a typical protocol would involve establishing subcutaneous tumor xenografts in immunodeficient mice (e.g., nude mice or NSG mice) using 786-O, SJSA-1, or A431 cells. When tumors reach 100-200 mm3, mice are randomized and treated with GPX4-IN-2 formulated in a suitable vehicle (e.g., 10% DMSO + 40% PEG300 + 5% Tween80 + 45% saline). 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, tumor weight, body weight, survival. Plasma and tumor tissue are collected for pharmacokinetic analysis and pharmacodynamic evaluation: GPX4 activity in tumor lysates, lipid peroxidation (MDA, 4-HNE), GSH/GSSG ratio, and GPX4 expression by Western blot. Histological analysis of tumors (H&E, Ki-67, TUNEL) and normal tissues (liver, kidney) for toxicity assessment. Ferroptosis can be confirmed by co-administration of ferrostatin-1 which should block the antitumor effect. This protocol is generic and not validated for GPX4-IN-2 specifically. |
| ADME/Pharmacokinetics |
No specific pharmacokinetic data for GPX4-IN-2. Based on its molecular properties (MW 444.65, LogP ~5-6 from structure), the compound is highly lipophilic, which suggests high plasma protein binding, extensive tissue distribution, and likely low aqueous solubility. Oral bioavailability may be poor, favoring intraperitoneal (IP) administration for in vivo studies. The compound contains an amide group that may be susceptible to hydrolysis. Metabolism would occur in the liver via phase I (oxidation, reduction) and phase II (glucuronidation, sulfation) enzymes. Plasma half-life, Cmax, AUC, clearance, volume of distribution, and oral bioavailability have not been publicly reported. Solubility: DMSO 37.5 mg/mL (84.34 mM). In vivo formulation: 10% DMSO + 40% PEG300 + 5% Tween80 + 45% saline (3.75 mg/mL suspension). Storage: 4degC, stored under nitrogen; in solvent at -80degC for 6 months.
|
| Toxicity/Toxicokinetics |
No specific toxicity data for GPX4-IN-2. 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 (liver, kidney, pancreas). GPX4 is essential for survival of certain cell types, and systemic GPX4 inhibition or knockout is lethal. Therefore, toxicity is expected at effective doses, and the therapeutic window may be narrow. In cell-based studies, GPX4-IN-2 shows antiproliferative activity against cancer cells. In vivo toxicity studies would assess maximum tolerated dose (MTD), body weight loss, organ toxicity (liver enzymes ALT/AST, kidney function BUN/creatinine), and histopathological changes in normal tissues. No data are publicly available. For research use only, handle with extreme caution. Safety Data Sheet (SDS) should be consulted before use.
|
| References | |
| Additional Infomation |
GPX4-IN-2 (Compound 28, CAS: 2485005-22-7) is a potent inhibitor of glutathione peroxidase 4 (GPX4), a key regulator of ferroptosis. The compound exhibits antiproliferative activity against cancer cells including 786-O (renal carcinoma), SJSA-1 (osteosarcoma), and A431 (epidermoid carcinoma), with IC50 values in the low micromolar range. GPX4-IN-2 is a research tool for studying ferroptosis, iron metabolism, and cancer biology. By blocking GPX4 activity, it prevents the reduction of lipid peroxides, leading to lipid peroxide accumulation and ferroptosis-linked cell death. Molecular formula: C30H40N2O, molecular weight: 444.65. Appearance: solid. Solubility: DMSO 37.5 mg/mL (84.34 mM). Storage: 4degC, stored under nitrogen; powder stable for 3 years at -20degC. Not approved for clinical use. For research use only. GPX4-IN-2 is an apoptosis/metabolic enzyme/protease pathway compound with potential in cancer research applications.
|
| Molecular Formula |
C30H40N2O
|
|---|---|
| Molecular Weight |
444.651408195496
|
| Exact Mass |
444.314
|
| CAS # |
2485005-22-7
|
| Related CAS # |
GPX4-IN-1
|
| PubChem CID |
155125294
|
| Appearance |
Off-white to light yellow solid powder
|
| LogP |
7.5
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
3
|
| Rotatable Bond Count |
7
|
| Heavy Atom Count |
33
|
| Complexity |
614
|
| Defined Atom Stereocenter Count |
2
|
| SMILES |
C12(NC3=CC=C([C@H]4C5=C(C=C(OC)C=C5)C[C@H](CCCC)N4)C=C3)CC3CC(CC(C3)C1)C2
|
| InChi Key |
YBBBVOMLRKBZDK-MLMUNBGCSA-N
|
| InChi Code |
InChI=1S/C30H40N2O/c1-3-4-5-26-15-24-16-27(33-2)10-11-28(24)29(31-26)23-6-8-25(9-7-23)32-30-17-20-12-21(18-30)14-22(13-20)19-30/h6-11,16,20-22,26,29,31-32H,3-5,12-15,17-19H2,1-2H3/t20?,21?,22?,26-,29-,30?/m0/s1
|
| Chemical Name |
N-[4-[(1S,3S)-3-butyl-6-methoxy-1,2,3,4-tetrahydroisoquinolin-1-yl]phenyl]adamantan-1-amine
|
| HS Tariff Code |
2934.99.9001
|
| 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 (e.g. under nitrogen), 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)
|
| Solubility (In Vitro) |
DMSO: ~100 mg/mL (~224.9 mM; with ultrasonication)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.62 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween-80 + 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 DMSO stock solution (25.0 mg/mL) to 400 μL of PEG300 and mix well; then add 50 μL of Tween-80 and mix well; finally add 450 μL of physiological saline and 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 (5.62 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), Suspended solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of DMSO stock solution (25.0 mg/mL) to 900 μL of 20% SBE-β-CD saline and mix well. 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (5.62 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 DMSO stock solution (25.0 mg/mL) to 900 μL of corn oil and mix well. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.2490 mL | 11.2448 mL | 22.4896 mL | |
| 5 mM | 0.4498 mL | 2.2490 mL | 4.4979 mL | |
| 10 mM | 0.2249 mL | 1.1245 mL | 2.2490 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
