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> Necrosulfonamide [1360614-48-7]

Necrosulfonamide [1360614-48-7]

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Referentie HY-100573-1mL

Formaat : 10mM/1mL

Merk : MedChemExpress


Description

Necrosulfonamide is a MLKL and Gasdermin D (GSDMD) inhibitor, capable of separately inhibiting necroptosis and pyroptosis of cells. Necrosulfonamide does not affect the activation of upstream signals, but specifically inhibits the downstream executor oligomerization step. Necrosulfonamide reduces the expression of the key kinases NLRP3 and caspase-1 involved in necroptosis and pyroptosis, activate the Nrf2 pathway and the downstream antioxidant enzymes, and also downregulates a variety of inflammatory factors. Necrosulfonamide plays significant roles in various diseases such as neurodegenerative diseases (such as Parkinson’s disease), tissue damage and ischemia-reperfusion injury, inflammatory bowel disease, osteoarthritis and fracture repair, and hair loss by regulating two important programmed necrosis pathways[1][2][3][4][5][6][7].

IC50 & Target[3][6]

NLRP3

 

Caspase-1

 

IL-6

 

IL-1β

 

iNOS

 

HO-1

 

Cellular Effect
Cell Line Type Value Description References
HT-29 IC50
<200 nM
Compound: 66; NSA
Anti-neprotic activity in human HT-29 cells assessed as reduction in TSZ-induced necroptosis incubated for 24 hrs by cell titer glo-based luminescence assay
Anti-neprotic activity in human HT-29 cells assessed as reduction in TSZ-induced necroptosis incubated for 24 hrs by cell titer glo-based luminescence assay
[PMID: 31622096]
HT-29 IC50
<1 μM
Compound: 34; NSA
Anti-necroptotic activity in human HT-29 cells assessed as inhibition of T/S/Z induced necroptosis by CellTiter-Glo Luminescent Cell Viability assay
Anti-necroptotic activity in human HT-29 cells assessed as inhibition of T/S/Z induced necroptosis by CellTiter-Glo Luminescent Cell Viability assay
[PMID: 36781172]
HT-29 IC50
124 nM
Compound: 34; NSA
Anti-necroptotic activity in human HT-29 cells assessed as inhibition of T/S/Z induced necroptosis by CellTiter-Glo Luminescent Cell Viability assay
Anti-necroptotic activity in human HT-29 cells assessed as inhibition of T/S/Z induced necroptosis by CellTiter-Glo Luminescent Cell Viability assay
[PMID: 36781172]
HT-29 EC50
447 nM
Compound: NSA; A1
Anti-necroptosis activity against TNFalpha/Smac mimetic/Z-VAD-fmk-induced human HT-29 cells measured after 24 hrs by CellTiter-Glo assay
Anti-necroptosis activity against TNFalpha/Smac mimetic/Z-VAD-fmk-induced human HT-29 cells measured after 24 hrs by CellTiter-Glo assay
[PMID: 38442525]
Jurkat IC50
<1 μM
Compound: 34; NSA
Anti-necroptotic activity in FADD null human Jurkat cells assessed as inhibition of T/S/Z induced necroptosis by CellTiter-Glo Luminescent Cell Viability assay
Anti-necroptotic activity in FADD null human Jurkat cells assessed as inhibition of T/S/Z induced necroptosis by CellTiter-Glo Luminescent Cell Viability assay
[PMID: 36781172]
In Vitro

Necrosulfonamide (0-10 μM) blocks necrosis in both human colon cancer HT-29 cells and FADD null human T cell leukemia Jurkat cells with IC50s both less than 1 μM, but it has no effect on mouse cells, for that the cysteine at the 86th position in human MLK1 is replaced by tryptophan in the mouse MLKL after covalent modification[1].
Necrosulfonamide (6 h) acts downstream of RIP3 activation, it does not inhibit the interaction or phosphorylation between RIP1 and RIP3; but enhances these processes in RIP3-HT-29 cells[1].
Necrosulfonamide (10 μM, 45 min-25 h) inhibits the release of inflammatory factors and pyroptosis in BMDMs and NCM460 cells necroptosis and has no effect on the vitality of the two types of cells[4].
Necrosulfonamide (0.1-100 μM) protects primary cultured astrocytes and human astrocytes against oxygen-glucose deprivation and reoxygenation (OGD/Re)-induced cell injury, reduces the number of PI-positive cells and inhibits the levels of necroptosis-related proteins[5].
Necrosulfonamide (0.1-1 μM, 6 h) reverses pyroptosis-induced inhibition of proliferation and differentiation of osteoblasts through the NLRP3/caspase-1/GSDMD pathway in hFOB 1.19 cells[6].

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

Necrosulfonamide Related Antibodies

ELISA Assay[4]

Cell Line: BMDMs
Concentration: 10 μM
Incubation Time: 1 h, followed by LPS for 6 h
Result: Inhibited the massive release of IL-1β and TNF-α.

Apoptosis Analysis[6]

Cell Line: hFOB 1.19 cells
Concentration: 0.1, 0.5, 1 μM
Incubation Time: 6 h
Result: Significantly decreased the pyroptosis rate at 0.5 μM and 1.0 μM.
Did not significantly change the viability or pyroptosis rate of osteoblasts alone.

RT-PCR[6]

Cell Line: hFOB 1.19 cells
Concentration: 0.1, 0.5, 1 μM
Incubation Time: 6 h
Result: Significantly decreased the pyroptosis rate at 0.5 μM and 1.0 μM.
Did not significantly change the viability or pyroptosis rate of osteoblasts alone.

Western Blot Analysis[6]

Cell Line: hFOB 1.19 cells
Concentration: 0.1, 0.5, 1 μM
Incubation Time: 6 h
Result: Inhibited the activation of the pyroptosis execution proteins (caspase-1, GSDMD).
Increased the protein expressions of key osteogenic markers (ALP, Runx2, COL-1, OPN, BMP-2)

ELISA Assay[6]

Cell Line: hFOB 1.19 cells
Concentration: 0.1, 0.5, 1 μM
Incubation Time: 6 h
Result: Reduced the levels of IL-6, TNF-α and IL-1β in the cell supernatant
In Vivo

Necrosulfonamide (20 mg/kg, i.p., single dose) alleviates Lipopolysaccharide (HY-D1056A1)/D-galactosamine (HY-42682)-induced acute liver failure in mice[2].
Necrosulfonamide (1-5 mg/kg, i.p., once daily for 3 days) reduces oxidative Stress, inflammation, and dopaminergic neuronal cell death in MPTP (HY-15608)-induced Parkinson’s disease mouse model[3].
Necrosulfonamide (20-40 mg/kg, i.p., once other daily for 5-7 days) ameliorates Inflammatory bowel disease (IBD) in mice via inhibiting GSDMD-medicated pyroptosis and MLKL-mediated necroptosis[4].
Necrosulfonamide (40-80 nmol, intracerebroventricular administration, single dose) produces neuroprotective effects against ischemia/reperfusion (I/R)-induced acute brain injury in rats[5].
Necrosulfonamide promotes hair growth and prevents hair follicle degeneration in androgenetic alopecia (AGA) mice through Wnt signaling[7].

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

Animal Model: LPS/D-galactosamine-induced acute liver failure established in male 8-week-old specific pathogen-free (SPF) grade C57BL/6J mice[2]
Dosage: 20 mg/kg
Administration: Intraperitoneal injection (i.p.), single dose before the model creation process
Result: Increased the survival rate of mice, reduced the serum ALT level, and alleviated liver tissue damage.
Reduced the levels of serum IL-1β and IL-18.
Downregulated the protein levels of NLRP3, cleaved caspase-1, cleaved caspase-11, and mature IL-1β.
Animal Model: MPTP-induced Parkinson’s disease mouse model established in male C57BL/6 mice (24-25 g, 9-10 weeks old)[3]
Dosage: 1 and 5 mg/kg
Administration: Intraperitoneal injection (i.p.), single dose before the model creation process
Result: Reduced the death of dopaminergic neurons and restores neurotrophic factors.
Reduced the levels of pro-inflammatory mediators such as iNOS, TNF-α, IL-1β, and IL-6.
Reduced the activation of microglial cells and astrocytes in the substantia nigra and striatum.
Restored the expression of the core antioxidant transcription factor Nrf2 and a series of downstream antioxidant enzymes (HO-1, catalase, MnSOD, GCLC, GCLM).
Effectively inhibited the expression of p-MLKL and MLKL, and reduced the number of p-MLKL+/TH+ and p-MLKL+/OX-42+ cells.
Animal Model: DSS (HY-116282C) induced IBD model established in male 6- to 9-week-old C57BL/6 mice weighting 22-26 g [4]
Dosage: 20 and 40 mg/kg (prevent) and 20 mg/kg (treatment)
Administration: Intraperitoneal injection (i.p.), once every other day for 7 days (prevent) and for 5 days (treatment)
Result: Significantly reduced the DAI score and increased the colon length in a dose-dependent manner.
Reduced the levels of serum TNF-α and MPO.
Reduced the inflammatory score, retained more goblet cells and mucus layers, and decreased the infiltration of CD4+ T cells, CD8+ T cells and macrophages (F4/80+) in the colon tissue.
Reduced the levels of 16sRNA, IL-1β, TNF-α, and IL-6 mRNA, inhibited p-p65, and promoted Nrf2 expression.
Reduced the death of colonic epithelial cells and p-MLKL-positive cells, and inhibited the expression of necroptosis markers (p-MLKL, p-RIPK1, p-RIPK3) and pyroptosis markers (N-GSDMD).
Animal Model: Transient middle cerebral artery occlusion model established in adult male SD rats (290-310 g)[5]
Dosage: 40 and 80 nmol
Administration: Intracerebroventricular administration, single dose
Result: Significantly reduced the volume of cerebral infarction.
Significantly improved the neurological function score and the asymmetry of forelimb usage.
Significantly reduced the number of PI-positive cells.
Inhibited the total expression levels of MLKL/p-MLKL, RIP3K/p-RIP3K, and RIP1K/p-RIP1K in the ischemic penumbra tissue, and translocation of MLKL/p-MLKL and RIP3K/p-RIP3K to the nucleus and nuclear membrane.
Masse moléculaire

461.47

Formule

C18H15N5O6S2
CAS No.
Appearance

Solid

Color

Light yellow to yellow

SMILES

O=C(NC1=CC=C(S(=O)(NC2=NC=CN=C2OC)=O)C=C1)/C=C/C3=CC=C([N+]([O-])=O)S3
Livraison

Room temperature in continental US; may vary elsewhere.
Stockage
Powder -20°C 3 years
4°C 2 years
In solvent -80°C 2 years
-20°C 1 year
Solvant et solubilité
In Vitro: 

DMSO : ≥ 28 mg/mL (60.68 mM; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

*"≥" means soluble, but saturation unknown.

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 2.1670 mL 10.8349 mL 21.6699 mL
5 mM 0.4334 mL 2.1670 mL 4.3340 mL
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

    Solubility: 2.5 mg/mL (5.42 mM); Suspended solution; Need ultrasonic

    This protocol yields a suspended solution of 2.5 mg/mL. Suspended solution can be used for oral and intraperitoneal injection.

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

    Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
  • Protocol 2

    Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in Saline)

    Solubility: 2.5 mg/mL (5.42 mM); Suspended solution; Need ultrasonic

    This protocol yields a suspended solution of 2.5 mg/mL. Suspended solution can be used for oral and intraperitoneal injection.

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

    Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.

For the following dissolution methods, please prepare the working solution directly. It is recommended to prepare fresh solutions and use them promptly within a short period of time.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  50% PEG300    50% Saline

    Solubility: 10 mg/mL (21.67 mM); Suspended solution; Need ultrasonic

  • Protocol 2

    Add each solvent one by one:  20% SBE-β-CD in Saline

    Solubility: 6.67 mg/mL (14.45 mM); Suspended solution; Need ultrasonic
Pureté et documentation
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