2. Apoptosis NF-κB Metabolic Enzyme/Protease Immunology/Inflammation MAPK/ERK Pathway
  3. Necroptosis Reactive Oxygen Species (ROS) Mitochondrial Metabolism Mixed Lineage Kinase RIP kinase
  4. Necroptosis inducer 2

Necroptosis inducer 2 is a necroptosis inducer and copper chelator. Necroptosis inducer 2 chelates intracellular free copper ions, disrupts redox homeostasis, elevates ROS levels, disrupts mitochondrial membrane potential, and induces cancer cell necroptosis. Necroptosis inducer 2 upregulates the necroptosis markers p-MLKL and p-RIP3 expression. Necroptosis inducer 2 exhibits anti-tumor activity in mice. Necroptosis inducer 2 can be used for the research of cancer, such as triple-negative breast cancer.

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Necroptosis inducer 2

Necroptosis inducer 2 Chemical Structure

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Description

Necroptosis inducer 2 is a necroptosis inducer and copper chelator. Necroptosis inducer 2 chelates intracellular free copper ions, disrupts redox homeostasis, elevates ROS levels, disrupts mitochondrial membrane potential, and induces cancer cell necroptosis. Necroptosis inducer 2 upregulates the necroptosis markers p-MLKL and p-RIP3 expression. Necroptosis inducer 2 exhibits anti-tumor activity in mice. Necroptosis inducer 2 can be used for the research of cancer, such as triple-negative breast cancer[1].

IC50 & Target[1]

RIPK3

 

In Vitro

Necroptosis inducer 2 (Compound 2d) (20 μM) binds selectively and strongly to Cu2+ and Cu+ in a cell-free system, with Ka values of 1.42 × 104/M and 7.33 × 103/M, respectively[1].
Necroptosis inducer 2 (48 h) potently inhibits the proliferation of MDA-MB-231, 4T1, HeLa, HT29, A2780, and U-2 OS cancer cells with IC50 values ranging from 4.1-10.6 μM, shows moderate activity against A549 cells (IC50 = 29.4 μM), and exhibits reduced toxicity toward non-cancerous MCF-10A cells (IC50 = 10.6 μM)[1].
Necroptosis inducer 2 (10 μM; 44 h treatment) has its cytotoxicity rescued in MDA-MB-231 cells by exogenous Cu2+, confirming copper chelation is essential for its antiproliferative activity[1].
Necroptosis inducer 2 (10-20 μM; 24 h) reduces intracellular chelatable copper levels in MDA-MB-231 cells in a concentration-dependent manner, as detected by CS1 confocal microscopy[1].
Necroptosis inducer 2 (20 μM; 24 h) rescues MDA-MB-231 cells from CuCl2-induced cytotoxicity, demonstrating potent copper-chelating activity in a cellular context[1].
Necroptosis inducer 2 (5-20 μM; 24 h) elevates intracellular ROS levels and disrupts mitochondrial membrane potential in MDA-MB-231 cells in vitro in a concentration-dependent manner, and this effect is reversed by exogenous Cu2+[1].
Necroptosis inducer 2 (10 μM; 44 h) induces cell death in MDA-MB-231 cells primarily via the necroptosis pathway, as only Necrostatin-1 (HY-15760) rescues cell viability[1].
Necroptosis inducer 2 (5-20 μM; 24 h) upregulates the necroptosis markers p-MLKL and p-RIP3 in MDA-MB-231 cells in vitro in a concentration-dependent manner[1].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Necroptosis inducer 2 Related Antibodies

Western Blot Analysis[1]

Cell Line: MDA-MB-231 cells
Concentration: 5 μM, 10 μM, 20 μM
Incubation Time: 24 h
Result: Caused a concentration-dependent upregulation of phosphorylated mixed lineage kinase domain-like (p-MLKL) and phosphorylated receptor-interacting protein kinase 3 (p-RIP3), key necroptosis markers.
Showed that markers for apoptosis, autophagy, and ferroptosis remained unchanged.

Immunofluorescence[1]

Cell Line: MDA-MB-231 cells
Concentration: 10 μM, 20 μM
Incubation Time: 24 h
Result: Caused a marked concentration-dependent increase in p-MLKL accumulation, providing direct visual evidence of necroptosis activation.
In Vivo

Necroptosis inducer 2 (Compound 2d) (5 mg/kg; intratumoral; 2 doses (day 1 and day 6 postmodeling)) inhibits murine breast tumor growth in vivo via induction of necroptosis, with no observable systemic toxicity[1].
Necroptosis inducer 2 (5-20 mg/kg; intratumoral; 2 doses (day 1 and day 8 postmodeling)) exerts dose-dependent in vivo murine breast tumor growth inhibition, reaching 89.8% at 20 mg/kg with complete regression in some animals and no detectable hematological or systemic toxicity[1].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Model: Breast cancer BALB/c mice (female, 4−6 weeks old, subcutaneous injection of 1 × 106 4T1 murine breast cancer cells)[1]
Dosage: 5 mg/kg
Administration: intratumoral; 2 doses (day 1 and day 6 postmodeling)
Result: Achieved a tumor inhibition rate of 36.3%.
Showed no significant body weight loss compared to the control group.
Revealed no obvious histopathological damage in major organs via H&E staining.
Induced extensive necrosis and structural disruption in tumor tissue via H&E staining.
Downregulated PCNA expression in tumor tissue via immunohistochemical analysis.
Caused a significant increase in p-MLKL staining in tumor tissue via immunohistochemical analysis.
Animal Model: Breast cancer BALB/c mice (female, 4−6 weeks old, subcutaneous injection of 1 × 106 4T1 murine breast cancer cells)[1]
Dosage: 5 mg/kg; 10 mg/kg; 20 mg/kg
Administration: intratumoral; 2 doses (day 1 and day 8 postmodeling)
Result: Inhibited tumor growth in a dose-dependent manner, with tumor inhibition rates of 35.3%, 58.8%, and 89.8% at 5, 10, and 20 mg/kg, respectively.
Induced complete tumor regression in 2 out of 5 mice in the 20 mg/kg group.
Showed no significant body weight changes across all treated groups.
Revealed no hematological toxicity compared to the control group via red blood cell and platelet parameter analysis.
Molecular Weight

1114.08

Formula

C72H56Cl2N2O2P2
SMILES

OC1=C(C=C(C=C1)C[P+](C2=CC=CC=C2)(C3=CC=CC=C3)C4=CC=CC=C4)/C=N/C5=C(C6=C(C=C5)C=CC=C6)C7=C8C(C=CC=C8)=CC=C7/N=C/C9=C(C=CC(C[P+](C%10=CC=CC=C%10)(C%11=CC=CC=C%11)C%12=CC=CC=C%12)=C9)O.[Cl-].[Cl-]
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Necroptosis inducer 2 Related Classifications

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Keywords:

Necroptosis inducer 2Necroptosis inducer2Necroptosis inducer-2NecroptosisReactive Oxygen Species (ROS)Mitochondrial MetabolismMixed Lineage KinaseRIP kinaseMLKsReceptor-interacting protein kinasesRIPKnecroptosismitochondrial damageovarian cancerosteosarcomacopper ionsreactive oxygen specieslung cancertriple-negative breast cancercolon cancerMDA-MB-231Inhibitorinhibitorinhibit

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