Genentech, Inc., South San Francisco, California (X.Liu., X.Lia., J.L., S.U., J.Chen, J.Cheng, T.L., J.L., J.N., S.S.-L., C.Q., E.S., M.W., C.E.C.A.H., T.P.H.) and QPS, Delaware Technology Park, Newark, Delaware (E.S.).
Genentech, Inc., South San Francisco, California (X.Liu., X.Lia., J.L., S.U., J.Chen, J.Cheng, T.L., J.L., J.N., S.S.-L., C.Q., E.S., M.W., C.E.C.A.H., T.P.H.) and QPS, Delaware Technology Park, Newark, Delaware (E.S.)
Drug Metab Dispos. 2020 May;48(5):408-419. doi: 10.1124/dmd.119.089763. Epub 2020 Mar 4.
The objectives of the present study were to characterize GNE-947 for its phosphoinositide 3-kinase (PI3K) and mammalian target of rapamycin (mTOR) inhibitory activities, in vitro anti-cell migration activity in human umbilical vein endothelial cells (HUVECs), in vivo antineovascularization activity in laser-induced rat choroidal neovascular (CNV) eyes, pharmacokinetics in rabbit plasma and eyes, and ocular distribution using matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) and autoradioluminography. Its PI3K and mTOR were 0.0005 and 0.045 µM, respectively, and its HUVEC IC was 0.093 µM. GNE-947 prevented neovascularization in the rat CNV model at 50 or 100 µg per eye with repeat dosing. After a single intravenous injection at 2.5 and 500 μg/kg in rabbits, its plasma terminal half-lives ( ) were 9.11 and 9.59 hours, respectively. After a single intravitreal injection of a solution at 2.5 μg per eye in rabbits, its apparent values were 14.4, 16.3, and 23.2 hours in the plasma, vitreous humor, and aqueous humor, respectively. After a single intravitreal injection of a suspension at 33.5, 100, 200 μg per eye in rabbits, the were 29, 74, and 219 days in the plasma and 46, 143, and 191 days in the eyes, respectively. MALDI-IMS and autoradioluminography images show that GNE-947 did not homogenously distribute in the vitreous humor and aggregated at the injection sites after injection of the suspension, which was responsible for the long of the suspension because of the slow dissolution process. This hypothesis was supported by pharmacokinetic modeling analyses. In conclusion, the PI3K/mTOR inhibitor GNE-947 prevented neovascularization in a rat CNV model, with up to approximately 6 months after a single intravitreal injection of the suspension in rabbit eyes. SIGNIFICANCE STATEMENT: GNE-947 is a potent phosphoinositide 3-kinase/mammalian target of rapamycin inhibitor and exhibits anti-choroidal neovascular activity in rat eyes. The duration of GNE-947 in the rabbit eyes after intravitreal injection in a solution is short, with a half-life ( ) of less than a day. However, the duration after intravitreal dose of a suspension is long, with up to 6 months due to low solubility and slow dissolution. These results indicate that intravitreal injection of a suspension for low-solubility drugs can be used to achieve long-term drug exposure.
本研究的目的是研究 GNE-947 的磷酸肌醇 3-激酶（PI3K）和哺乳动物雷帕霉素靶蛋白（mTOR）抑制活性，在人脐静脉内皮细胞（HUVEC）中的体外抗细胞迁移活性，在激光诱导的大鼠脉络膜新生血管（CNV）眼中的体内抗血管生成活性，在兔血浆和眼中的药代动力学，以及使用基质辅助激光解吸/电离成像质谱（MALDI-IMS）和放射自显影进行眼内分布。其 PI3K 和 mTOR 的 IC50 分别为 0.0005 和 0.045 µM，HUVEC 的 IC50 为 0.093 µM。GNE-947 在重复给药时，以每只眼 50 或 100 µg 的剂量预防大鼠 CNV 模型中的新生血管形成。在兔中以 2.5 和 500 µg/kg 静脉内单次注射后，其血浆终末半衰期（）分别为 9.11 和 9.59 小时。在兔中以每只眼 2.5 µg 的溶液进行单次玻璃体内注射后，其在血浆、玻璃体液和房水中的表观分布容积（）分别为 14.4、16.3 和 23.2 小时。在兔中以每只眼 33.5、100 和 200 µg 的悬浮液进行单次玻璃体内注射后，其在血浆和眼中的分别为 29、74 和 219 天和 46、143 和 191 天。MALDI-IMS 和放射自显影图像显示，GNE-947 在用悬浮液注射后不会均匀分布在玻璃体液中，并且在注射部位聚集，这导致了悬浮液的长，因为其溶解过程缓慢。该假设得到了药代动力学建模分析的支持。总之，PI3K/mTOR 抑制剂 GNE-947 可预防大鼠 CNV 模型中的新生血管形成，在兔眼玻璃体内注射悬浮液后，可达约 6 个月。意义陈述：GNE-947 是一种有效的磷酸肌醇 3-激酶/哺乳动物雷帕霉素靶蛋白抑制剂，在大鼠眼中具有抗脉络膜新生血管活性。GNE-947 在兔眼玻璃体内注射溶液后的半衰期（）很短，不到一天。然而，由于溶解度低和溶解缓慢，混悬剂给药后的时间很长，可达 6 个月。这些结果表明，低溶解度药物的玻璃体内注射混悬剂可用于实现长期药物暴露。
