Necrostatin-1

Names

[ Name ]:
Necrostatin-1

[Synonym ]:
5-(1H-Indol-3-ylmethyl)-3-methyl-2-thioxo-4-imidazolidinone
Necrostatin-1
5-(1H-Indol-3-ylmethyl)-3-methyl-2-thioxoimidazolidin-4-one
Necrostatin 1
MTH-trp
4-Imidazolidinone, 5-(1H-indol-3-ylmethyl)-3-methyl-2-thioxo-
5-indol-3-ylmethyl-3-methyl-2-thioxo-imidazolidin-4-one
Nec-1
Necroptotic Inhibitor,Nec-1
MTH-DL-Tryptophan
methyl-thiohydantoin-tryptophan

Biological Activity

[Description]:

Necrostatin-1 is a potent, selective and cell-permeable necroptosis inhibitor with an EC50 of 490 nM in Jurkat cells. It acts by inhibiting the death domain kinase RIP (RIP1) in the necroptosis pathway.

[Related Catalog]:

Signaling Pathways >> Autophagy >> Autophagy

Signaling Pathways >> Apoptosis >> RIP kinase

Research Areas >> Cancer

[Target]

RIP1 kinase[1]

[In Vitro]

Necrostatin-1 (Nec-1) is a specific and potent small-molecule inhibitor of cell death caused by death-domain receptor (DR) stimulation in the presence of caspase inhibition in multiple cell types. Necrostatin-1 efficiently inhibits the TNFα-induced necrotic death of L929 cells, which does not require exogenous caspase inhibitors[1]. Necrostatin-1 (Nec-1) prevents radiocontrast media (RCM)-induced dilation of peritubular capillaries, suggesting a novel role unrelated to cell death for the RIP1 kinase domain in the regulation of microvascular hemodynamics and pathophysiology of contrast-induced AKI (CIAKI)[2]. The decreased viability of C6 glioma cells caused by 3.0 µM and 6.0 µM shikonin is improved by pretreatment with Necrostatin-1 (Nec-1) to 92.3% and 82.9% at 1.5 h and 84.4% and 78.6% at 3.0 h, respectively. Similarly, the viability of U87 glioma cells is elevated by Necrostatin-1 to 91.6% and 81.5% at 1.5 h, and 81.8% and 71.2% at 3.0 h, respectively[3]. Necrostatin-1 (Nec-1) (30 µM) increases the survival of cardiomyocyte progenitor cell (CMPCs) by inhibiting necrotic cell death[4].

[In Vivo]

Necrostatin-1 (Nec-1) induces tubular bilation and affects the kinetics of the dilation of peritubular capillaries after RCM application. Upon a single intraperitoneal application of a single dose of Necrostatin-1 (1.65 mg/kg body weight, i.p.) 15 minutes before RCM, the return to baseline levels is prevented within the observation period[2].

[Cell Assay]

C6 (3×105 cells/well) and U87 (1.5×105 cells/well) glioma cells are seeded onto 96-well microplate and cultured 24 h. PBS is added into the control group and Shikonin is added into experimental group to reach the final concentration. Cellular viability is assessed using an MTT assay after Shikonin treatment at indicated time point. The absorbance value (A) at 570 nm is read using an automatic multi-well spectrophotometer. Two groups of glioma cells from the same cell line are treated with Shikonin at lower or higher concentration, respectively; other two groups of glioma cells are treated 1 h with 100 µM Necrostatin-1 or 40 µM z-VAD-fmk prior to co-incubation with Shikonin at indicated concentration. Additionally, another two groups of glioma cells are treated only with 100 µM Necrostatin-1 or 40 µM Z-VAD-fmk at corresponding time point[3].

[Animal admin]

Mice[2] 8-10 week old male C57BL/6 mice (average weight approx.23 g) are used. Mice receive intravenous application of 200 μL PBS or radiocontrast media (RCM) via the tail vein. A single dose of Z-VAD-fmk (10 mg/kg body weight) or Necrostatin-1 (1.65 mg/kg body weight) is applied intraperitoneally 15 min. before RCM-injection. Mice are harvested another 24 hours after RCM-application (48 hours after reperfusion). Blood samples are obtained from retroorbital bleeding and serum levels of urea and creatinine are determined.

[References]

[1]. Degterev A, et al. Chemical inhibitor of nonapoptotic cell death with therapeutic potential for ischemic brain injury. Nat Chem Biol. 2005 Jul;1(2):112-9.

[2]. Linkermann A, et al. The RIP1-kinase inhibitor necrostatin-1 prevents osmotic nephrosis and contrast-induced AKI in mice. J Am Soc Nephrol. 2013 Oct;24(10):1545-57.

[3]. Huang C, et al. Shikonin kills glioma cells through necroptosis mediated by RIP-1. PLoS One. 2013 Jun 28;8(6):e66326.

[4]. Feyen D, et al. Increasing short-term cardiomyocyte progenitor cell (CMPC) survival by necrostatin-1 did not further preserve cardiac function. Cardiovasc Res. 2013 Jul 1;99(1):83-91.

[5]. Zhou K, et al. RIP1-RIP3-DRP1 pathway regulates NLRP3 inflammasome activation following subarachnoid hemorrhage. Exp Neurol. 2017 Sep;295:116-124.

[Related Small Molecules]

GSK872 | GSK2982772 | WEHI-345 | RIPA-56 | GSK 583 | RIP2 kinase inhibitor 1 | GSK'481 | RIP2 kinase inhibitor 2 | Nec-4

Chemical & Physical Properties

[ Density]:
1.4±0.1 g/cm3

[ Boiling Point ]:
441.9±37.0 °C at 760 mmHg

[ Melting Point ]:
151ºC

[ Molecular Formula ]:
C₁₃H₁₃N₃OS

[ Molecular Weight ]:
259.327

[ Flash Point ]:
221.1±26.5 °C

[ Exact Mass ]:
259.077942

[ PSA ]:
80.22000

[ LogP ]:
1.26

[ Appearance of Characters ]:
solid

[ Vapour Pressure ]:
0.0±1.1 mmHg at 25°C

[ Index of Refraction ]:
1.738

[ Storage condition ]:
−20°C

[ Water Solubility ]:
DMSO: >10 mg/mL

MSDS

Click to get detail MSDS

Safety Information

[ Personal Protective Equipment ]:
Eyeshields;Gloves;type N95 (US);type P1 (EN143) respirator filter

[ Risk Phrases ]:
36/37/39

[ Safety Phrases ]:
36/37/39

[ RIDADR ]:
NONH for all modes of transport

[ WGK Germany ]:
3

[ HS Code ]:
2933990090

Synthetic Route

Click to get detail synthetic route

Precursor & DownStream

Precursor

DownStream

Customs

[ HS Code ]: 2933990090

[ Summary ]:
2933990090. heterocyclic compounds with nitrogen hetero-atom(s) only. VAT:17.0%. Tax rebate rate:13.0%. . MFN tariff:6.5%. General tariff:20.0%

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