| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| 100mg | |||
| 250mg | |||
| 500mg | |||
| Other Sizes |
| Targets |
NecroX-5 targets mitochondrial reactive oxygen species (ROS) and peroxynitrite (ONOO⁻), acting as a potent scavenger of these reactive species. By reducing mitochondrial ROS levels, the compound prevents oxidative stress-induced cell death and necrosis. NecroX-5 also reduces intracellular calcium concentration by inhibiting Ca²⁺ influx, which leads to decreased mitochondrial ROS levels and inhibition of cell migration. The compound modulates AKT signaling, as NecroX-5-mediated reduction in mitoROS is associated with AKT downregulation. These pleiotropic mechanisms—ROS scavenging, calcium modulation, and AKT signaling—contribute to its anti-inflammatory, anti-cancer, and cytoprotective effects. The compound does not target a specific receptor or enzyme but rather acts through the modulation of cellular redox state and calcium homeostasis.
|
|---|---|
| ln Vitro |
Breast cancer cell migration is inhibited by necroX-5 (10 or 40 μM), which is mediated by AKT inhibition and linked to decreased mitoROS. By inhibiting Ca2+ inflow, NecroX-5 (10 μM) lowers the intracellular calcium concentration of 4T1 cells, which in turn results in decreased levels of mitochondrial ROS, cell migration inhibition, and AKT downregulation [1]. In LPS-stimulated H9C2 cells, a 24-hour treatment with NecroX-5 (10 µM) can effectively suppress the elevated production of TNFα, TGFβ1, pSmad2, and Dcn [2].
In vitro, NecroX-5 (10 or 40 μM) inhibits breast cancer cell migration, which is mediated by AKT inhibition and associated with reduced mitochondrial ROS levels. By inhibiting Ca²⁺ influx, NecroX-5 (10 μM) decreases intracellular calcium concentration in 4T1 cells, leading to reduced mitochondrial ROS levels, inhibition of cell migration, and AKT downregulation. In LPS-stimulated H9C2 cells, NecroX-5 protects against inflammatory damage. The compound has been shown to protect cells from oxidative stress, hypoxia, and cold shock in vitro. These cytoprotective effects are attributed to its ROS scavenging activity and its ability to modulate calcium signaling. NecroX-5's anti-cancer effects are further supported by its ability to reduce breast cancer cell migration. The compound is a valuable tool for studying the roles of oxidative stress and mitochondrial dysfunction in cell death and disease. |
| ln Vivo |
In tumor-bearing TUBO-P2J mice, NecroX-5 (2.5 mg/kg, every other day) suppresses the spread of breast cancer cells [1].
In vivo, NecroX-5 protects against carbon tetrachloride (CCl₄)-induced liver injury and chronic liver fibrosis in rodent models. In these models, administration of NecroX-5 reduces liver damage, as measured by serum transaminase levels and histopathological examination. The compound's hepatoprotective effects are attributed to its ability to scavenge ROS and ONOO⁻ generated during CCl₄ metabolism. NecroX-5 also exhibits anti-inflammatory effects in various disease models. The compound's ability to reduce intracellular calcium and mitochondrial ROS levels contributes to its protective effects in vivo. These findings support the potential therapeutic application of NecroX-5 in diseases associated with oxidative stress, inflammation, and fibrosis. Further studies are needed to fully characterize its in vivo efficacy and safety profile. |
| Enzyme Assay |
In vitro enzyme/receptor binding assays are not applicable to NecroX-5 mesylate as it acts primarily as a ROS scavenger and calcium modulator rather than a conventional enzyme inhibitor or receptor antagonist. However, the compound's antioxidant activity can be quantified using cell-free assays. A typical protocol: the ROS scavenging activity of NecroX-5 is measured using the DCFH-DA assay or the ABTS/DPPH radical scavenging assay. For DCFH-DA, the compound is incubated with 2',7'-dichlorofluorescin diacetate (10 μM) in assay buffer (PBS, pH 7.4) for 30 minutes at 37°C. Fluorescence is measured at excitation 485 nm and emission 530 nm. For ONOO⁻ scavenging, peroxynitrite is generated from 3-morpholinosydnonimine (SIN-1), and the oxidation of dihydrorhodamine 123 is monitored fluorometrically. The compound's ability to inhibit necrosis is assessed in cell-free systems by measuring the release of lactate dehydrogenase (LDH) or the uptake of propidium iodide in the presence of necrotic stimuli. Each assay is performed in triplicate, and IC₅₀ values are calculated from dose-response curves.
|
| Cell Assay |
In vitro cell-based assays for NecroX-5 are performed using various cell lines to assess its cytoprotective and anti-cancer effects. A typical protocol: cells (e.g., 4T1 breast cancer cells, H9C2 cardiomyocytes, or primary hepatocytes) are seeded in 96-well plates at 10,000-20,000 cells/well and allowed to adhere overnight. Cells are pre-treated with NecroX-5 at concentrations ranging from 1 to 40 μM for 1-4 hours, then exposed to necrotic stimuli such as hydrogen peroxide (100-500 μM), tert-butyl hydroperoxide (t-BHP, 100-500 μM), or LPS (1 μg/mL). Cell viability is assessed using MTT, CellTiter-Glo, or LDH release assays. ROS levels are measured using DCFH-DA or MitoSOX staining followed by flow cytometry or fluorescence microscopy. Intracellular calcium levels are measured using Fluo-4 or Fura-2 AM. Cell migration is assessed using wound healing or Transwell assays. Each condition is tested in triplicate, and experiments are repeated at least three times.
|
| Animal Protocol |
In vivo animal studies for NecroX-5 are conducted in rodent models of liver injury, fibrosis, and other diseases. A typical protocol for CCl₄-induced liver injury: male C57BL/6 mice are administered CCl₄ (0.5-1.0 mL/kg, intraperitoneal) in olive oil (1:4) twice weekly for 4-8 weeks to induce liver fibrosis. NecroX-5 is administered via intraperitoneal injection at doses of 5-20 mg/kg, daily or every other day, starting before or concurrently with CCl₄ treatment. At study termination, serum is collected for ALT and AST measurements, and liver tissues are harvested for histopathological examination (H&E and Masson's trichrome staining) and biomarker analysis (e.g., α-SMA, collagen I by IHC or Western blot). For acute liver injury models, a single dose of CCl₄ is administered, and NecroX-5 is given 1-2 hours before or after CCl₄. Liver injury is assessed 24-48 hours later.
|
| ADME/Pharmacokinetics |
Pharmacokinetic properties of NecroX-5 mesylate have not been fully characterized in published literature. The compound is soluble in DMF (10 mg/mL), DMSO (1 mg/mL), and ethanol (0.25 mg/mL), suggesting moderate lipophilicity. It is supplied as a crystalline solid and stored at -20°C for long-term stability. The compound's plasma half-life, volume of distribution, oral bioavailability, and protein binding remain unknown. Based on its physicochemical properties, NecroX-5 is expected to have moderate to good membrane permeability, consistent with its cell-permeable nature. The mesylate salt form enhances aqueous solubility compared to the free base. Further pharmacokinetic studies would be required to determine its absorption, distribution, metabolism, and excretion profile. The compound is for research use only and has not been developed for clinical applications.
|
| Toxicity/Toxicokinetics |
Toxicological data for NecroX-5 mesylate are limited to observations from in vivo efficacy studies. At the doses used in preclinical studies (5-20 mg/kg, IP), NecroX-5 is generally well-tolerated, with no significant adverse effects reported. The compound has not undergone formal toxicology testing for regulatory purposes. Standard laboratory safety precautions should be followed when handling NecroX-5 mesylate: use of personal protective equipment (gloves, safety goggles, lab coat) and handling in a well-ventilated fume hood. The compound should be stored at -20°C for long-term stability. No genotoxicity, carcinogenicity, or reproductive toxicity data are available. Researchers should consult the safety data sheet (SDS) before handling. The compound is for research use only and is not approved for human use.
|
| References |
|
| Additional Infomation |
Additional information for NecroX-5 mesylate: The compound has a CAS number of 1383718-29-3. Its molecular formula is C₂₅H₃₁N₃O₃S • 2CH₄O₃S and molecular weight is 645.8 g/mol. Purity is typically ≥85-98%. Synonyms include NecroX-5, Necrox-5 methanesulfonate, and necrosis inhibitor 5. The compound is a cell-permeable inhibitor of necrosis with strong mitochondrial ROS and ONOO⁻ scavenging activity. It protects cells from oxidative stress, hypoxia, and cold shock in vitro. It protects against CCl₄-induced liver injury and chronic liver fibrosis in rodent models. This product is for research use only and is not approved for clinical applications. No FDA approvals or investigational new drug applications exist.
|
| Molecular Formula |
C27H39N3O9S3
|
|---|---|
| Molecular Weight |
645.808264017105
|
| Exact Mass |
645.184
|
| CAS # |
1383718-29-3
|
| Related CAS # |
1120332-90-2;1526790-45-3 (HCl);1383718-29-3 (mesylate);1526790-48-6 (HBr); 1526790-50-0 (sulfate);
|
| PubChem CID |
71514788
|
| Appearance |
Light brown to brown solid powder
|
| Hydrogen Bond Donor Count |
4
|
| Hydrogen Bond Acceptor Count |
11
|
| Rotatable Bond Count |
6
|
| Heavy Atom Count |
42
|
| Complexity |
778
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CS(=O)(=O)O.CS(=O)(=O)O.C1COCCC1CNC2=CC(=CC3=C2NC(=C3)C4=CC=CC=C4)CN5CCS(=O)(=O)CC5
|
| InChi Key |
ZWWWDIWEZYRVMB-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C25H31N3O3S.2CH4O3S/c29-32(30)12-8-28(9-13-32)18-20-14-22-16-23(21-4-2-1-3-5-21)27-25(22)24(15-20)26-17-19-6-10-31-11-7-19;2*1-5(2,3)4/h1-5,14-16,19,26-27H,6-13,17-18H2;2*1H3,(H,2,3,4)
|
| Chemical Name |
5-[(1,1-dioxo-1,4-thiazinan-4-yl)methyl]-N-(oxan-4-ylmethyl)-2-phenyl-1H-indol-7-amine;methanesulfonic acid
|
| 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 : ~170 mg/mL (~263.24 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.83 mg/mL (4.38 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 28.3 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: ≥ 2.83 mg/mL (4.38 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 28.3 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 | 1.5484 mL | 7.7422 mL | 15.4844 mL | |
| 5 mM | 0.3097 mL | 1.5484 mL | 3.0969 mL | |
| 10 mM | 0.1548 mL | 0.7742 mL | 1.5484 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
