GI 254023X is a selective inhibitor with an IC50 value of 5.3nM for MMP9 (Matrix Metalloproteinase 9). The selectivity of GI 254023X for ADAM10 (A Disintegrin and Metalloproteinase 10) is 100 times higher than that for ADAM17, with an IC50 value of 2.5nM. GI 254023X is primarily used in the research of immunology, inflammation, and neurodegenerative diseases[1]. GI 254023X selectively binds to the catalytic domains of ADAM10 and MMP9 to inhibit their enzymatic activities[2]. GI 254023X also prevents the degradation of E-cadherin in epithelial cells, which is crucial for maintaining cell-to-cell adhesion and tissue integrity[3]. GI 254023X reduces the degradation of extracellular matrix components, thereby affecting processes such as tissue remodeling and angiogenesis[4].
In vitro, pretreatment of PAC1/RAGE cells with GI 254023X (100nM - 25mM) for 1 hour followed by stimulation with 300nM PACAP-27 for 2 hours significantly inhibited PACAP-induced RAGE shedding. The inhibition rate reached 86% at 25mM and 75% at 1mM, with a slight inhibitory effect even at the low concentration of 100nM. At a concentration of 1mM, GI 254023X inhibited basal RAGE shedding by 67%[5]. In ovarian cancer cell lines A2780 and primary ovarian cancer cells, treatment with GI 254023X (100µM) significantly inhibited the release of CXCL16 (CXC chemokine ligand 16), increased the levels of CXCL16 on the cell membrane, and reduced the release of soluble CXCL16 (sCXCL16) in the culture supernatant, with an inhibition rate of up to 89.7%. GI 254023X also significantly inhibited the migratory capacity of ovarian cancer cells[6].
In vivo, GI 254023X (100mg/kg) was administered via intraperitoneal injection to mice 30 minutes and 24 hours after traumatic brain injury (TBI). GI 254023X significantly reduced brain tissue lesion volume in TBI mice, decreased the generation of the axonal injury marker SMI32, attenuated the upregulation of TNFα, IL-6, and LCN2, and increased the expression of the T cell marker Cd247[7]. GI 254023X (200mg/kg) was administered via intraperitoneal injection to 35-week-old PSAPP transgenic mice (an Alzheimer’s disease model) for 5 consecutive days. This treatment significantly reduced the levels of LRP1 in the brain and increased Aβ40 levels in the plasma[8].
References:
[1] Hoettecke N, Ludwig A, Foro S, et al. Improved synthesis of ADAM10 inhibitor GI254023X. Neurodegener Dis. 2010;7(4):232-8.
[2] Ma S, Xu J, Wang X, et al. Effect of ADAM10 Inhibitor GI254023X on Proliferation and Apoptosis of Acute T-Lymphoblastic Leukemia Jurkat Cells In Vitro and Its Possible Mechanisms. Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2015 Aug;23(4):950-5.
[3] Li J, Ouyang Q, Chen CW, et al. Neuron-Derived ADAM10 Production Stimulates Peripheral Nerve Injury-Induced Neuropathic Pain by Cleavage of E-Cadherin in Satellite Glial Cells. Pain Med. 2017 Sep 1;18(9):1752-1766.
[4] Kim YH, Jung JC. Suppression of tunicamycin-induced CD44v6 ectodomain shedding and apoptosis is correlated with temporal expression patterns of active ADAM10, MMP-9 and MMP-13 proteins in Caki-2 renal carcinoma cells. Oncol Rep. 2012 Nov;28(5):1869-74.
[5] Metz VV, Kojro E, Rat D, et al. Induction of RAGE shedding by activation of G protein-coupled receptors. PLoS One. 2012;7(7):e41823.
[6] Gooden MJ, Wiersma VR, Boerma A, et al. Elevated serum CXCL16 is an independent predictor of poor survival in ovarian cancer and may reflect pro-metastatic ADAM protease activity. Br J Cancer. 2014 Mar 18;110(6):1535-44.
[7] Appel D, Hummel R, Weidemeier M, et al. Pharmacologic Inhibition of ADAM10 Attenuates Brain Tissue Loss, Axonal Injury and Pro-inflammatory Gene Expression Following Traumatic Brain Injury in Mice. Front Cell Dev Biol. 2021 Mar 15;9:661462.
[8] Shackleton B, Crawford F, Bachmeier C. Inhibition of ADAM10 promotes the clearance of Aβ across the BBB by reducing LRP1 ectodomain shedding. Fluids Barriers CNS. 2016 Aug 8;13(1):14.
GI 254023X是一种选择性抑制剂，针对MMP9（Matrix Metalloproteinase 9）的IC50 值为5.3nM，GI 254023X对ADAM10（A Disintegrin and Metalloproteinase 10）的选择性是ADAM17的100倍，IC50 值为2.5nM。GI 254023X主要用于免疫学、炎症和神经退行性疾病的研究[1]。GI 254023X 通过选择性结合 ADAM10和MMP9的催化结构域，抑制它们的酶活性[2]。GI 254023X 还能够防止上皮细胞中E-cadherin的降解，这对于维持细胞间黏附和组织完整性非常重要[3]。在MMP9方面，GI 254023X的抑制作用减少了细胞外基质成分的降解，从而影响组织重塑和血管生成等过程[4]。
在体外，GI 254023X（100nM-25mM）预处理PAC1/RAGE细胞1小时，随后加入300nM PACAP-27刺激2小时，显著抑制了PACAP诱导的RAGE脱落，25mM浓度下抑制率达86%，1mM浓度下抑制约75%，即使在100nM低浓度下也有轻微抑制效果。GI 254023X在1mM浓度下对基础RAGE脱落的抑制率达67%[5]。在卵巢癌细胞系A2780和原代卵巢癌细胞中，GI 254023X（100µM）处理显著抑制了CXC趋化因子配体16（CXCL16）的释放，增加了细胞膜上CXCL16的水平，并减少了培养上清中可溶性CXCL16（sCXCL16）的释放，抑制率高达89.7%，GI 254023X还显著抑制了卵巢癌细胞的迁移能力[6]。
在体内，GI 254023X（100mg/kg）通过腹腔注射在小鼠创伤性脑损伤（TBI）后30分钟和24小时给药。GI 254023X显著减少了TBI小鼠脑组织损伤体积，降低了轴突损伤标志物SMI32的生成，并减轻了TNFα、IL-6和LCN2的上调，同时增加了T细胞标志物Cd247的表达[7]。在体内，GI 254023X（200mg/kg）通过腹腔注射在35周龄的PSAPP转基因小鼠（阿尔兹海默病模型），连续5天，显著降低了脑中LRP1的产生水平，同时增加了血浆中Aβ40水平[8]。