School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, P.O. Box 1627, FI-70211, Kuopio, Finland.
Institute of Biomedicine, Faculty of Health Sciences, University of Eastern Finland, P.O. Box 1627, FI-70211, Kuopio, Finland.
Sci Rep. 2019 Sep 6;9(1):12860. doi: 10.1038/s41598-019-49009-z.
L-Type Amino Acid Transporter 1 (LAT1/Lat1) is responsible for carrying large, neutral L-amino acids as well as several drugs and prodrugs across the blood-brain barrier (BBB). However, the BBB is not the only barrier that hinders drugs acting effectively within the brain; the brain parenchymal cell membranes represent a secondary barrier for the drugs with intracellular target sites. In this study, expression and function of Lat1 was quantified in mouse primary neuron, astrocyte and immortalized microglia (BV2) cultures. Moreover, ability of Lat1 to carry prodrugs inside these brain cells was evaluated. The results showed that Lat1 was localized at the similar level in all studied cells (3.07 ± 0.92-3.77 ± 0.91 fmol/µg protein). The transporter was also functional in all three cell types, astrocytes having the highest transport capacity and affinity for the LAT1/Lat1-substrate, [C]-L-leucine, followed by neurons and microglia. The designed prodrugs (1-6) were able to utilize Lat1 for their cellular uptake and it was mainly much higher than the one of their parent drugs. Interestingly, improved cellular uptake was also achieved in cells representing Alzheimer's Disease phenotype. Therefore, improved delivery and intra-brain targeting of drugs can be attained by utilizing LAT1/Lat1 and prodrug approach.
L 型氨基酸转运蛋白 1（LAT1/Lat1）负责将大的、中性的 L 型氨基酸以及几种药物和前体药物穿过血脑屏障（BBB）。然而，BBB 并不是唯一阻碍药物在大脑内有效作用的屏障；脑实质细胞膜是具有细胞内靶位的药物的第二个屏障。在这项研究中，在小鼠原代神经元、星形胶质细胞和永生化小胶质细胞（BV2）培养物中定量表达和功能 Lat1。此外，还评估了 Lat1 将前体药物带入这些脑细胞的能力。结果表明，Lat1 在所有研究的细胞中定位于相似的水平（3.07±0.92-3.77±0.91 fmol/µg 蛋白）。该转运体在所有三种细胞类型中均具有功能，星形胶质细胞对 LAT1/Lat1-底物[C]-L-亮氨酸的转运能力和亲和力最高，其次是神经元和小胶质细胞。设计的前体药物（1-6）能够利用 Lat1 进行细胞摄取，其摄取量明显高于其母体药物。有趣的是，在代表阿尔茨海默病表型的细胞中也实现了改善的细胞摄取。因此，通过利用 LAT1/Lat1 和前体药物方法，可以实现药物的递增强和脑内靶向。
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