GSK484 is a selective PAD4 inhibitor, has been used as an anti-ischemic compound and exerts inhibitory activity against PAD4. In the absence of calcium the IC50 value is 50 nM[1-3].
GSK484(25 nM; 1 h before irradiation) facilitated Triple-negative breast cancer (TNBC) cell apoptosis[4]. GSK484(100 nM; 15min) treatment promoted the radiosensitivity of CRC cells and induced cell death by promoting DNA double-strand breaks[5].
GSK484(4 mg/kg; i.p;40days) significantly inhibited tumor size, tumor weight and tumor volume in mice following irradiation in Nasopharyngeal carcinoma (NPC) mice model[6]. GSK484(4 mg/kg; i.p;3days) inhibits PAD4 to reduce distal lung injury by reducing neutrophil infiltration, neutrophil extracellular trap formation, apoptosis, and inflammatory cytokine secretion in mice ischemia reperfusion (IR) model[7].
参考文献:
[1]. Perdomo J, Leung HHL, et,al. Neutrophil activation and NETosis are the major drivers of thrombosis in heparin-induced thrombocytopenia. Nat Commun. 2019 Mar 21;10(1):1322. doi: 10.1038/s41467-019-09160-7. PMID: 30899022; PMCID: PMC6428879.
[2]. Perdomo J, Leung HHL, et,al. Neutrophil activation and NETosis are the major drivers of thrombosis in heparin-induced thrombocytopenia. Nat Commun. 2019 Mar 21;10(1):1322. doi: 10.1038/s41467-019-09160-7. PMID: 30899022; PMCID: PMC6428879.
[3]. Du M, Yang W, et,al. Inhibition of peptidyl arginine deiminase-4 protects against myocardial infarction induced cardiac dysfunction. Int Immunopharmacol. 2020 Jan;78:106055. doi: 10.1016/j.intimp.2019.106055. Epub 2019 Dec 6. PMID: 31816575.
[4]. Wei L, Wang X, et,al. The PAD4 inhibitor GSK484 enhances the radiosensitivity of triple-negative breast cancer. Hum Exp Toxicol. 2021 Jul;40(7):1074-1083. doi: 10.1177/0960327120979028. Epub 2020 Dec 23. Erratum in: Hum Exp Toxicol. 2021 Nov;40(11):2022. PMID: 33355008.
[5]. Wang B, Su X, et,al. GSK484, an inhibitor of peptidyl arginine deiminase 4, increases the radiosensitivity of colorectal cancer and inhibits neutrophil extracellular traps. J Gene Med. 2023 Sep;25(9):e3530. doi: 10.1002/jgm.3530. Epub 2023 May 19. PMID: 37203323.
[6]. Chen H, Luo M, et,al. Inhibition of PAD4 enhances radiosensitivity and inhibits aggressive phenotypes of nasopharyngeal carcinoma cells. Cell Mol Biol Lett. 2021 Mar 16;26(1):9. doi: 10.1186/s11658-021-00251-2. Erratum in: Cell Mol Biol Lett. 2021 Dec 30;26(1):55. Erratum in: Cell Mol Biol Lett. 2023 Apr 4;28(1):29. PMID: 33726680; PMCID: PMC7962337.
[7]. Du M, Yang L, et,al. Inhibition of Peptidyl Arginine Deiminase-4 Prevents Renal Ischemia-Reperfusion-Induced Remote Lung Injury. Mediators Inflamm. 2020 Dec 29;2020:1724206. doi: 10.1155/2020/1724206. PMID: 33456369; PMCID: PMC7787741.
GSK484是一种选择性PAD4抑制剂，已被用作抗缺血化合物，对PAD4具有抑制活性。不含钙时IC50为50 nM[1-3]。
GSK484(25 nM; 1 h before irradiation)促进三阴性乳腺癌(TNBC)细胞凋亡[4]。GSK484(100 nM; 15min)通过促进DNA双链断裂，提高CRC细胞的放射敏感性，诱导细胞死亡[5]。
GSK484(4 mg/kg; i.p;40days)显著抑制鼻咽癌(NPC)小鼠辐照后的肿瘤大小、肿瘤重量和肿瘤体积[6]。在小鼠缺血再灌注(IR)模型中，GSK484(4 mg/kg; i.p;3days)通过减少中性粒细胞浸润、中性粒细胞胞外陷阱形成、细胞凋亡和炎性细胞因子分泌，抑制PAD4减轻远端肺损伤[7]。
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Fluorescence Polarization (FP) binding affinity studies [1]: |
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Preparation method |
PAD4 was serially diluted in the presence of 10 nM GSK484 in Assay Buffer (100 mM HEPES, pH 8, 50 mM NaCl, 5% glycerol, 1 mM CHAPS, 1 mM DTT) at varying concentrations of calcium (0 mM, 0.2 mM, 2 mM and 10 mM). Following incubation for 50 min, apparent Kd values for each calcium concentration were determined using a single site saturation curve. For IC50 determination, test compounds were serially diluted in DMSO (1% final assay concentration) and tested at the same range of calcium concentrations in the presence of PAD4 (at the calculated Kd for each calcium condition) and 10 nM GSK484 in the same assay buffer and volume. Reactions were incubated for 50 min, after which IC50 values were calculated. |
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Applications |
GSK484 demonstrated high affinity binding to the low-calcium form of PAD4 with IC50 50 nM(no Ca+). |
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Cell experiment [2]: |
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Cell lines |
MDA-MB-231 cells (Triple-negative breast cancer (TNBC)) |
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Preparation method |
GSK484 was dissolved in DMSO. The DMSO concentration of the solution used was consistently lower than 0.1%. Cells were grown to 70% -80% confluence on plates and cultivated with a low dose of GSK484 for 1 h before irradiation. |
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Reaction Conditions |
25 nM; 1 h before irradiation |
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Applications |
GSK484 facilitates cell apoptosis and promotes the radiosensitivity of MDA-MB-231 cells. |
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Animal experiment [2]: |
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Animal models |
Female BALB/c mice (6-week-old) |
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Preparation method |
MDA-MB-231 cells were injected subcutaneously into the 4th mammary fat pads of lactiferous ducts the in mice. Mice were randomly divided into four groups (Vehicle, 15 Gy alone, GSK484 4 mg/kg, and 15 Gy + GSK484 4 mg/kg). GSK484 was administered at a dose of 4 mg/kg ip in 2-hydroxypropyl-β-cyclodextrin at 1 hour before irradiation. The tumor areas were irradiated with 6 MV X-rays (15 Gy in one fraction) in 15 Gy alone and 15 Gy + GSK484 4 mg/kg groups. |
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Dosage form |
4 mg/kg; i.p.; 1 hour before irradiation |
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Applications |
Treatment with GSK484 and ischemia reperfusion (IR), the tumor volume and body weight of mice decreased significantly. |
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参考文献: [1]. Lewis HD, Liddle J, et,al. Inhibition of PAD4 activity is sufficient to disrupt mouse and human NET formation. Nat Chem Biol. 2015 Mar;11(3):189-91. doi: 10.1038/nchembio.1735. Epub 2015 Jan 26. PMID: 25622091; PMCID: PMC4397581. |
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