Necrostatin-1

Necrostatin-1 is a novel small molecule inhibitor of necroptosis. Necrostatin-1 inhibits RIP1 kinase activity (EC₅₀=182nM) through allosteric binding. Necrostatin-1 inhibits necroptosis by blocking necrosome assembly and MLKL-mediated cell membrane rupture. Necrostatin-1 reduces inflammation release by inhibiting IDO. Necrostatin-1 can be used in research on cerebral ischemia-reperfusion injury, neurodegenerative diseases, inflammation, and tumors^[1-4].

In vitro, Necrostatin-1 (20μM) pretreated NRK-52E rat renal tubular epithelial cells for 24h, followed by treatment with TNF-α (10ng/mL) combined with antimycin A (10μM) for 24h. Necrostatin-1 significantly reduced cell death, increased cell viability, and decreased Drp1 protein expression^[5]. Necrostatin-1 (1-40μM) pretreated human neuroblastoma SH-SY5Y cells and mouse hippocampal HT-22 cells for 30min, followed by treatment with hydrogen peroxide (0.25-0.5mM)/6-hydroxydopamine (0.1-0.2mM) on SH-SY5Y cells for 24h. Necrostatin-1 significantly attenuated hydrogen peroxide/6-hydroxydopamine-induced cell death and increased cell viability^[6].

In vivo, Necrostatin-1 (0.0468mg/kg) was injected intra-articularly into C57BL/6 mice with a traumatic osteoarthritis model, twice a week for 8 weeks. Necrostatin-1 significantly alleviated knee OA cartilage degeneration in the model mice, reduced OARSI scores, and downregulated the expression of MMP3, MMP13, ADAMTs5, HMGB1, and TLR4 in cartilage tissue, and decreased serum SDF-1α levels^[7]. Necrostatin-1 (1μg; 50μL; single injection) was administered intracerebroventricularly to C57BL/6 mice with an intracerebral hemorrhage model, 1h before modeling. Necrostatin-1 significantly reduced the expression of RIP1, RIP3, MLKL, p-RIP3, and p-MLKL in the periventricular tissue of the model mice. Necrostatin-1 inhibited cell necrosis and microglial activation. Necrostatin-1 downregulated the expression of interleukin-1β, interleukin-6, and tumor necrosis factor-α^[8].

References:
[1] Cao L, Mu W. Necrostatin-1 and necroptosis inhibition: Pathophysiology and therapeutic implications. Pharmacol Res. 2021 Jan;163:105297.
[2] Chen KQ, Wang SZ, Lei HB, et al. Necrostatin-1: a promising compound for neurological disorders. Front Cell Neurosci. 2024 Jun 27;18:1408364.
[3] Cao Y, Wang HB, Ni CJ, et al. Necrostatin-1 prevents skeletal muscle ischemia reperfusion injury by regulating Bok-mediated apoptosis. J Chin Med Assoc. 2023 Jan 1;86(1):26-33.
[4] Yu C, Wang X, Qin J. Effect of necrostatin-1 on sciatic nerve crush injury in rat models. J Orthop Surg Res. 2023 Jan 30;18(1):74.
[5] Zhang L, Jiang F, Chen Y, et al. Necrostatin-1 attenuates ischemia injury induced cell death in rat tubular cell line NRK-52E through decreased Drp1 expression. Int J Mol Sci. 2013 Dec 18;14(12):24742-54.
[6] Jantas D, Chwastek J, Grygier B, et al. Neuroprotective Effects of Necrostatin-1 Against Oxidative Stress-Induced Cell Damage: an Involvement of Cathepsin D Inhibition. Neurotox Res. 2020 Mar;37(3):525-542.
[7] Liang S, Lv ZT, Zhang JM, et al. Necrostatin-1 Attenuates Trauma-Induced Mouse Osteoarthritis and IL-1β Induced Apoptosis via HMGB1/TLR4/SDF-1 in Primary Mouse Chondrocytes. Front Pharmacol. 2018 Nov 27;9:1378.
[8] Liu C, Cao Y, Wang HX, et al. Necrostatin-1 decreases necroptosis and inflammatory markers after intraventricular hemorrhage in mice. Neural Regen Res. 2022 Dec;17(12):2710-2716.