糖尿病视网膜病变是糖尿病的继发性微血管并发症。该疾病从非增殖性和增殖性糖尿病视网膜病变两个阶段进展，后者的特征是视网膜异常血管生成。视网膜血管生成的药理学管理采用昂贵且侵入性的玻璃体内注射生物药物（抗血管内皮生长因子药物）。为了寻找能够充当抗血管生成剂的小分子，我们将研究重点放在阿西替尼上，它是一种酪氨酸激酶抑制剂，代表肾细胞癌的二线治疗方法。阿西替尼是血管内皮生长因子受体的抑制剂，其中酪氨酸激酶抑制剂（舒尼替尼和索拉非尼）对血管内皮生长因子受体1和2最具选择性。除了众所周知的抗血管生成和免疫调节功能外，我们特此通过生物信息学/分子建模方法和糖尿病视网膜病变的体外模型探索了阿西替尼的多药理学特征。我们通过用高葡萄糖浓度（波动和非波动）挑战视网膜内皮细胞，在两种不同的糖尿病视网膜病变体外模型中展示了阿西替尼的抗血管生成活性。我们发现，阿西替尼可抑制血管内皮生长因子受体 1（1.82 ± 0.10；0.54 ± 0.13，波动高血糖与阿西替尼 1 µM 中的磷酸化蛋白水平分别）和血管内皮生长因子受体 2（2.38 ± 0.21） ；0.98 ± 0.20，波动高血糖中的磷酸化蛋白水平与阿西替尼 1 µM 相比，能够显着降低 (p < 0.05) Nrf2 的表达（1.43 ± 0.04；0.85 ± 0.01，波动高血糖中的蛋白水平）通过预测的 Keap1 相互作用和黑皮质素受体 1 的激活，在暴露于高葡萄糖的视网膜内皮细胞中分别与阿西替尼 1 µM 相比。此外，阿西替尼治疗显着 (p < 0.05) 降低了活性氧的产生 (0.90 ± 0.10; 0.44 ± . 0.06，高葡萄糖与阿西替尼 1 µM 中的荧光单位，分别）和抑制 ERK 通路（1.64 ± 0.09；0.73 ± 0.06，波动高葡萄糖与 . axitinib 1 µM，分别）在暴露于高葡萄糖的 HREC 中。所获得的关于新兴多药理学特征的结果支持这样的假设：阿西替尼可能是治疗糖尿病视网膜病变的有效候选者，具有辅助作用机制。版权所有 © 2024 Lazzara、Conti、Sasmal、Alikunju、Rossi、Drago、Platania 和 Bucolo。

Diabetic retinopathy is a secondary microvascular complication of diabetes mellitus. This disease progresses from two stages, non-proliferative and proliferative diabetic retinopathy, the latter characterized by retinal abnormal angiogenesis. Pharmacological management of retinal angiogenesis employs expensive and invasive intravitreal injections of biologic drugs (anti-vascular endothelial growth factor agents). To search small molecules able to act as anti-angiogenic agents, we focused our study on axitinib, which is a tyrosine kinase inhibitor and represents the second line treatment for renal cell carcinoma. Axitinib is an inhibitor of vascular endothelial growth factor receptors, and among the others tyrosine kinase inhibitors (sunitinib and sorafenib) is the most selective towards vascular endothelial growth factor receptors 1 and 2. Besides the well-known anti-angiogenic and immune-modulatory functions, we hereby explored the polypharmacological profile of axitinib, through a bioinformatic/molecular modeling approach and in vitro models of diabetic retinopathy. We showed the anti-angiogenic activity of axitinib in two different in vitro models of diabetic retinopathy, by challenging retinal endothelial cells with high glucose concentration (fluctuating and non-fluctuating). We found that axitinib, along with inhibition of vascular endothelial growth factor receptors 1 (1.82 ± 0.10; 0.54 ± 0.13, phosphorylated protein levels in fluctuating high glucose vs . axitinib 1 µM, respectively) and vascular endothelial growth factor receptors 2 (2.38 ± 0.21; 0.98 ± 0.20, phosphorylated protein levels in fluctuating high glucose vs . axitinib 1 µM, respectively), was able to significantly reduce (p < 0.05) the expression of Nrf2 (1.43 ± 0.04; 0.85 ± 0.01, protein levels in fluctuating high glucose vs . axitinib 1 µM, respectively) in retinal endothelial cells exposed to high glucose, through predicted Keap1 interaction and activation of melanocortin receptor 1. Furthermore, axitinib treatment significantly (p < 0.05) decreased reactive oxygen species production (0.90 ± 0.10; 0.44 ± 0.06, fluorescence units in high glucose vs . axitinib 1 µM, respectively) and inhibited ERK pathway (1.64 ± 0.09; 0.73 ± 0.06, phosphorylated protein levels in fluctuating high glucose vs . axitinib 1 µM, respectively) in HRECs exposed to high glucose. The obtained results about the emerging polypharmacological profile support the hypothesis that axitinib could be a valid candidate to handle diabetic retinopathy, with ancillary mechanisms of action.Copyright © 2024 Lazzara, Conti, Sasmal, Alikunju, Rossi, Drago, Platania and Bucolo.