| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
| Targets |
RIPK1
Necrostatin-2 racemate targets receptor-interacting protein kinase 1 (RIPK1), a key regulator of necroptosis and inflammatory signaling. RIPK1 is involved in the activation of NF-κB and the induction of apoptosis and necroptosis. By inhibiting RIPK1 autophosphorylation, Necrostatin-2 racemate blocks necroptosis, a programmed form of necrotic cell death. The compound shows high selectivity for RIPK1 over other kinases. It does not inhibit IDO (indoleamine 2,3-dioxygenase). |
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| ln Vitro |
Nec-1s is a potent RIPK1 inhibitor and cellular necroptosis while lacking IDO inhibitory activity[1].
In vitro, Necrostatin-2 racemate potently inhibits necroptosis with an EC₅₀ of 50 nM. It prevents RIPK1 autophosphorylation. The compound shows >1000-fold selectivity for RIPK1 over 485 other human kinases. It does not inhibit IDO. In cellular assays, Necrostatin-2 racemate blocks TNF-α-induced necroptosis in various cell lines. Detailed in vitro data are available in the primary literature. |
| ln Vivo |
Nec-1s is effective at minimizing brain injuries. It is a superior inhibitor that can be used in vivo and does not have a paradoxical sensitizing effect in TNF-induced lethality[1].
Nec-1s differs from Nec-1 in that it has a number of favorable pharmacokinetic and pharmacodynamic properties. Because of their lower in vivo and in vitro toxicity, Nec-1s have a safer safety profile than Nec-1[2]. In vivo data for Necrostatin-2 racemate are limited in publicly available sources. Based on its potent and specific RIPK1 inhibition, the compound is expected to be useful for in vivo studies of necroptosis in various disease models including ischemic injury, neurodegeneration, and inflammatory diseases. Further studies are needed to fully characterize its in vivo efficacy and pharmacokinetic profile. The compound is a research tool for cell death studies. |
| Enzyme Assay |
Necrostatin 2 (also known as Necrostatin-2, Necrostatin-2 racemate, (±)-Necrostatin-2, 7-Cl-O-Nec-1 and Nec-1s, is a potent necroptosis inhibitor with EC50 of 50 nM.
In vitro kinase assays for Necrostatin-2 racemate typically use recombinant RIPK1 enzyme and a peptide or protein substrate. The compound is dissolved in DMSO and diluted in assay buffer (50 mM HEPES, pH 7.5, 10 mM MgCl₂, 2 mM DTT). Reactions are initiated by adding ATP and incubated at 30°C for 30-60 minutes. Phosphorylated substrate is detected by fluorescence polarization, AlphaScreen, or radiometric methods. IC₅₀ and EC₅₀ values are calculated from dose-response curves. Selectivity profiling is performed against a panel of kinases. Autophosphorylation is assessed by Western blotting. |
| Cell Assay |
Cells were treated with indicated concentrations of drug.
For cell-based assays, cell lines (e.g., Jurkat, U937, or L929) are cultured in RPMI-1640 or DMEM with 10% FBS. Cells are seeded in 96-well plates and pre-treated with Necrostatin-2 racemate at various concentrations (0.01-10 μM) for 30-60 minutes. Necroptosis is induced by TNF-α plus z-VAD-fmk (caspase inhibitor) with or without SMAC mimetic. Cell death is measured by propidium iodide uptake, LDH release, or MTT assay. RIPK1 phosphorylation is assessed by Western blotting. All treatments include vehicle controls and are performed in triplicate. |
| Animal Protocol |
In vivo, Necrostatin-2 racemate may be administered to rodents via intraperitoneal (IP) or intravenous (IV) injection. The compound is formulated in a suitable vehicle such as DMSO, PEG, or saline-based solutions. For disease models (e.g., ischemic injury, neurodegeneration, inflammatory diseases), animals are dosed at various regimens. Endpoints include tissue damage assessment, inflammatory cytokine levels, and survival analysis. Blood and tissue samples are collected for pharmacokinetic and pharmacodynamic analysis. All procedures follow institutional animal care guidelines.
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| ADME/Pharmacokinetics |
Necrostatin-2 racemate (MW 277.70, C₁₃H₁₂ClN₃O₂) is soluble in DMSO (≥50 mg/mL). The compound is a more stable form of Necrostatin-1. It shows >1000-fold selectivity for RIPK1 over 485 other kinases. Detailed pharmacokinetic parameters are not extensively reported in publicly available sources. The compound is typically stored at -20°C. Further ADME studies are needed to fully characterize its pharmacokinetic profile.
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| Toxicity/Toxicokinetics |
Toxicology data for Necrostatin-2 racemate are limited in publicly available sources. As a kinase inhibitor, potential off-target effects should be considered. The compound is for research use only and not intended for human therapeutic applications. Standard safety pharmacology and toxicology studies would be required for therapeutic development. The compound should be handled with standard laboratory safety precautions.
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| References | |
| Additional Infomation |
Necrostatin-2 racemate (Nec-1s) is a research-grade RIPK1 inhibitor for studying necroptosis and cell death pathways. Its primary applications include cell death research, immunology, and drug discovery. The compound is not approved for clinical use and has not entered clinical trials. It is commercially available from various chemical suppliers for research purposes only. Its mechanism involves inhibition of RIPK1 autophosphorylation, blocking necroptosis without affecting apoptosis or IDO activity.
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| Molecular Formula |
C13H12CLN3O2
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|---|---|
| Molecular Weight |
277.70628
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| Exact Mass |
277.061
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| Elemental Analysis |
C, 56.23; H, 4.36; Cl, 12.77; N, 15.13; O, 11.52
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| CAS # |
852391-15-2
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| Related CAS # |
Necrostatin 2;852391-19-6;Necrostatin 2 S enantiomer;852391-20-9
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| PubChem CID |
643953
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| Appearance |
White to light yellow solid powder
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| Density |
1.5±0.1 g/cm3
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| Index of Refraction |
1.676
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| LogP |
1.76
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
2
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
19
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| Complexity |
403
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O=C1N(C)C(=O)C(CC2C3C(=C(C=CC=3)Cl)NC=2)N1
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| InChi Key |
WIKGAEMMNQTUGL-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C13H12ClN3O2/c1-17-12(18)10(16-13(17)19)5-7-6-15-11-8(7)3-2-4-9(11)14/h2-4,6,10,15H,5H2,1H3,(H,16,19)
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| Chemical Name |
5-[(7-chloro-1H-indol-3-yl)methyl]-3-methylimidazolidine-2,4-dione
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| Synonyms |
7-Cl-O-Nec-1; Nec-1s; (±)-Necrostatin-2; Necrostatin-2
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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) |
DMSO: ≥ 50 mg/mL (~180.0 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 3 mg/mL (10.80 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 30.0 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: ≥ 3 mg/mL (10.80 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 30.0 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. View More
Solubility in Formulation 3: ≥ 3 mg/mL (10.80 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 | 3.6009 mL | 18.0044 mL | 36.0088 mL | |
| 5 mM | 0.7202 mL | 3.6009 mL | 7.2018 mL | |
| 10 mM | 0.3601 mL | 1.8004 mL | 3.6009 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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