Necrosulfonamide

Necrosulfonamide (NSA) is a specific inhibitor of MLKL (mixed lineage kinase domain-like protein)^[1]. Necrosulfonamide, an inhibitor of GSDMD, could inhibit PPVI-induced activation of the NLRP3 inflammasome^[2].

In vitro efficacy test it shown that IC₅₀ values for Necrosulfonamide determined in cell-based necroptosis (NEC), apoptosis (APOP), or toxicity (TOX) assays in Jurkat and U937 cells were 1399 nM, 6197 nM, 454 nM, >100,000 nM and 14694 nM, respectively^[3]. In vitro, treatment with 10 µM or 20 µM Necrosulfonamide inhibited GSDMD-mediated IL-1β release after inflammasome stimulation, demonstrating complete suppression of IL-1β even 60 min after stimulation in cells stimulated with LPS and nigericin^[4].

In vivo, male adult C57BL/6 mice were administrated 5 mg/kg Necrosulfonamide intraperitoneally twice a day significantly reduced the hematoma size, suppressed inflammatory cells and cytokines and protected the blood-brain barrier compared to vehicle controls^[1]. In vivo, rats were injected with Necrosulfonamide (10 mg/kg) reduced the levels of NOD-like receptor 3 (NLRP3), GSDMD-N, phosphorylated-MLKL, and phosphorylated-RIP3 levels in cardiac tissue with corresponding reductions in inflammatory cytokine levels^[5]. In vivo, mice were injected 1, 5, or 10 mg/kg Necrosulfonamide intraperitoneally improved the motor function and spinal edema of SCI-Mice with a therapeutic window^[6]. In vivo, rats were administrated with 0.5 mg/body Necrosulfonamide intraperitoneally protected lung IRI through the inhibition of necroptosis^[7]. Mice were treated with NSA (0.5 mg/kg/day for 3 days and 1 mg/kg/day for 7 days, every second day of i.p. injection) restorted motor performance defects, striatal TH + fiber deficiency, and TH + cell loss in a mouse model of PD^[8]. In vivo, rats were administrated with 1.65 mg/kg/day Necrosulfonamide intraperitoneally for 6 weeks obviously amended AlCl3-induced spatial learning and memory deficits, as demonstrated by enhanced rat performance in Morris water and Y-mazes^[9].

References:

[1] Zhang X, et al. Necrosulfonamide Alleviates Acute Brain Injury of Intracerebral Hemorrhage via Inhibiting Inflammation and Necroptosis. Front Mol Neurosci. 2022 Jun 2;15:916249.

[2] Teng JF, et al. Polyphyllin VI Induces Caspase-1-Mediated Pyroptosis via the Induction of ROS/NF-κB/NLRP3/GSDMD Signal Axis in Non-Small Cell Lung Cancer. Cancers (Basel). 2020 Jan 13;12(1):193.

[3] RÜbbelke M, et al. Locking mixed-lineage kinase domain-like protein in its auto-inhibited state prevents necroptosis. Proc Natl Acad Sci U S A. 2020 Dec 29;117(52):33272-33281.

[4] Rathkey JK, et al. Chemical disruption of the pyroptotic pore-forming protein gasdermin D inhibits inflammatory cell death and sepsis. Sci Immunol. 2018 Aug 24;3(26):eaat2738.

[5] He F, et al. Necrosulfonamide improves post-resuscitation myocardial dysfunction via inhibiting pyroptosis and necroptosis in a rat model of cardiac arrest. Eur J Pharmacol. 2022 Jul 5;926:175037.

[6] Jiao J, et al. Necrosulfonamide Ameliorates Neurological Impairment in Spinal Cord Injury by Improving Antioxidative Capacity. Front Pharmacol. 2020 Jan 9;10:1538.

[7] Ueda S, et al. Protective effect of necrosulfonamide on rat pulmonary ischemia-reperfusion injury via inhibition of necroptosis. J Thorac Cardiovasc Surg. 2022 Feb;163(2):e113-e122.

[8] Leem YH, et al. Necrosulfonamide exerts neuroprotective effect by inhibiting necroptosis, neuroinflammation, and α-synuclein oligomerization in a subacute MPTP mouse model of Parkinson's disease. Sci Rep. 2023 May 31;13(1):8783.

[9] Motawi TMK, et al. Ameliorative Effect of Necrosulfonamide in a Rat Model of Alzheimer's Disease: Targeting Mixed Lineage Kinase Domain-like Protein-Mediated Necroptosis. ACS Chem Neurosci. 2020 Oct 21;11(20):3386-3397.

Necrosulfonamide(NSA)是MLKL(混合系激酶域样蛋白)的一种特异性抑制剂^[1]。Necrosulfonamide是GSDMD的抑制剂,可以抑制PPVI诱导的NLRP3炎症体的激活^[2]。

体外药效试验表明,在Jurkat和U937细胞中,在基于细胞的坏死(NEC)、凋亡(APOP)或毒性(TOX)试验中确定的Necrosulfonamide的IC₅₀值分别为1399 nM、6197 nM、454 nM、>100000 nM和14694 nM^[3]。在体外,用10µM或20µM Necrosulfonamide处理可抑制炎症体刺激后GSDMD介导的IL-1β释放,在用LPS和nigericin刺激的细胞中,甚至在刺激后60分钟就显示出完全抑制IL-1β^[4]。

体内试验表明,雄性成年C57BL/6小鼠腹腔注射5mg/kg Necrosulfonamide,每天两次,与对照组相比,明显减少血肿大小,抑制炎症细胞和细胞因子,保护血脑屏障^[1]。在体内,给大鼠注射Necrosulfonamide(10mg/kg),可降低心脏组织中NOD样受体3(NLRP3)、GSDMD-N、磷酸化MLKL和磷酸化RIP3的水平,并相应降低炎症细胞因子水平^[5]。在体内,小鼠腹腔注射1、5或10 mg/kg Necrosulfonamide,可改善SCI小鼠的运动功能和脊髓水肿,具有治疗作用^[6]。体内试验表明,大鼠腹腔注射0.5 mg/kg Necrosulfonamide,通过抑制坏死,保护肺部IRI^[7]。用NSA治疗小鼠(0.5 mg/kg/d,连续3天,1 mg/kg/d,连续7天,每隔一天静脉注射一次)可恢复PD小鼠模型的运动表现缺陷、纹状体TH+纤维缺乏和TH+细胞损失^[8]。在体内,大鼠腹腔注射1.65 mg/kg/d的Necrosulfonamide,持续6周,明显修正了AlCl3诱导的空间学习和记忆缺陷,这表现在大鼠在Morris水和Y-mazes中的表现增强^[9]。