Background
SU5416 is a potent small molecule vascular endothelial growth factor receptor (VEGFR) inhibitor. SU5416 is a 3-substituted indolin-2-one compound with relatively high specificity for VEGFR-2 and VEGFR-1, used extensively in animal models of PH, primarily due to effects on pulmonary vascular endothelial cell apoptosis and proliferation.[1] SU416 has been developed for the treatment of solid human tumors as well. [3]
In vitro study was performed to examine the inhibitory effect of SU5416 on KDR phosphorylation. Which indicated that pretreatment of BCECs with SU5416 resulted in a dose-dependent inhibition of KDR phosphorylation with an IC50 of 0.29 ± 0.071 μM (n=6) SU5416 almost completely inhibited KDR phosphorylation at the concentration of 3 μM. Few BCECs were stained with trypan blue after the treatment of SU5416, at least up to the concentration of 3 μM for 24 h. This suggested that the inhibitory effect of SU5416 on KDR phosphorylation was not due to the cell toxicity.[2]
In vivo study demonstrated that SU5416 could significantly reverse LPS-induced ALI in mice, and exert better protective effect in TLR4 knockout mice. SU5416 could also act as a protective agent against LPS-induced ALI in mice. Moreover, SU5416 dramatically restored the reduction of CD31 expression mediated by LPS, suggesting SU5416 could rescue LPS-induced dysfunction of pulmonary endothelial barrier. In addition, both p-VEGFR2 and VEGFR2 expressions were inhibited by SU5416 in WT and TLR4?/- mice. SU5416 could attenuate LPS-induced ALI through modulating the VEGF/VEGFR and NF-κB pathways, which suggested SU5416 might be used for the treatment of patients with inflammation-mediated ALI.[3]
参考文献:
[1]. Peloquin GL, et al. SU5416 does not attenuate early RV angiogenesis in the murine chronic hypoxia PH model. Respir Res. 2019 Jun 17;20(1):123.
[2] Takeda A, et al. Suppression of experimental choroidal neovascularization utilizing KDR selective receptor tyrosine kinase inhibitor. Graefes Arch Clin Exp Ophthalmol. 2003 Sep;241(9):765-72.
[3]. Huang X, et al. SU5416 attenuated lipopolysaccharide-induced acute lung injury in mice by modulating properties of vascular endothelial cells. Drug Des Devel Ther. 2019 May 23;13:1763-1772.
SU5416 是一种有效的小分子血管内皮生长因子受体 (VEGFR) 抑制剂。 SU5416 是一种 3-取代的二氢吲哚-2-酮化合物，对 VEGFR-2 和 VEGFR-1 具有相对较高的特异性，广泛用于 PH 动物模型，主要是由于对肺血管内皮细胞凋亡和增殖的影响。 [1] SU416 也已开发用于治疗实体人类肿瘤。 [3]
进行了体外研究以检查 SU5416 对 KDR 磷酸化的抑制作用。这表明用 SU5416 预处理 BCECs 导致 KDR 磷酸化的剂量依赖性抑制，IC50 为 0.29 ± 0.071 μM (n=6) SU5416 在 3 μM 的浓度下几乎完全抑制 KDR 磷酸化。在 SU5416 处理后，很少有 BCEC 被台盼蓝染色，至少达到 3 μM 的浓度 24 小时。这表明SU5416对KDR磷酸化的抑制作用不是由于细胞毒性。[2]
体内研究表明，SU5416 可显着逆转 LPS 诱导的小鼠 ALI，并对 TLR4 基因敲除小鼠发挥更好的保护作用。 SU5416 还可以作为 LPS 诱导的小鼠 ALI 的保护剂。此外，SU5416 显着恢复了 LPS 介导的 CD31 表达降低，表明 SU5416 可以挽救 LPS 诱导的肺内皮屏障功能障碍。此外，在 WT 和 TLR4-/- 小鼠中，p-VEGFR2 和 VEGFR2 表达均被 SU5416 抑制。 SU5416可通过调节VEGF/VEGFR和NF-κB通路减轻LPS诱导的ALI，提示SU5416可用于治疗炎症介导的ALI。[3]
Protocol
| Cell experiment [1]: |
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Cell lines
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Human microvascular endothelial cells of the lung-blood (HMVEC-LBl)
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Preparation Method
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Cells were plated into 24-well plates and grown to confluence in EGM-2MV medium containing 2.5% FBS plus supplements. Only cells in passage 5 were used, with n = 12 wells per experimental condition.
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Reaction Conditions
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HMVEC-LBl was exposed to human VEGF-121 (40 ng/mL) in-vitro in serum-free medium for 7 h, in the absence or presence of the VEGF receptor antagonist, SU5416 (3 and 10 μM).
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Applications
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SU5416 can block the effects of VEGF and completely prevent VEGFR2 phosphorylation as well. There was no evidence of background VEGFR2 phosphorylation in the HMVEC-LBl and in serum-free medium. Adding SU5416 did not affect the background phosphorylation. In the absence of VEGF, SU5416 increased ET-1 production, by 16% at 10 μM, and SU5416 can completely abolish the VEGF effect on ET-1 production.
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| Animal experiment [2]: |
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Animal models
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Wild-type C57BL/6 mice (male, 8–10 weeks, 20–24 g) and genetically engineered TLR4-de?cient mice (male, 8–10 weeks, 20–22 g)
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Preparation Method
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Mice were stimulated by intratracheal administration of LPS after anesthetization with an intraperitoneal (i.p.) injection of tribromoethanol. Isopyknic saline-treated mice served as blank control group. After LPS stimulation, the experimental mice were treated with SU5416, DXM or DXM + SU5416 by oral administration for 12 hours. DXM-treated mice were served as positive control group.
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Dosage form
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20 mg/kg, BW solution in DMSO
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Applications
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SU5416 could be implemented to suppress immune response in mice with ALI. Normally, excessive activation and penetration of neutrophils is the common pathological process in LPS-induced ALI. Which subsequently enhanced the release of proinflammatory cytokines, which can further aggravate lung injury. SU5416 exhibited inhibitory effect on the population of neutrophil cell (P
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参考文献:
[1]. Star GP, et al. Effects of vascular endothelial growth factor on endothelin-1 production by human lung microvascular endothelial cells in vitro. Life Sci. 2014 Nov 24;118(2):191-4.
[2]. Huang X, et al. SU5416 attenuated lipopolysaccharide-induced acute lung injury in mice by modulating properties of vascular endothelial cells. Drug Des Devel Ther. 2019 May 23;13:1763-1772.
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