Aim: Galantamine is an acetylcholinesterase inhibitor frequently used in Alzheimer’s disease management. Its cholinergic adverse effects and rapid elimination limit its therapeutic outcomes. We inv...

L-Type amino acid transporter 1 (LAT1), expressed abundantly in the brain and placenta and overexpressed in several cancer cell types, has gained a lot of interest in drug research and development, as it can be utilized for brain-targeted drug delivery, as well as inhibiting the essential amino acid supply to cancer cells. The structure of LAT1 is today very well-known and the interactions of ligands at the binding site of LAT1 can be modeled and explained. However, less is known of LAT1′s life cycle within the cells. Moreover, the functionality of LAT1 can be measured by several different methods, which may vary between the laboratories and make the comparison of the results challenging. In the present study, the usefulness of indirect cis-inhibition methods and direct cellular uptake methods and their variations to interpret the interactions of LAT1-ligands were evaluated. Moreover, this study also highlights the importance of understanding the intracellular kinetics of LAT1-ligands, and how they can affect the regular function of LAT1 in critical tissues, such as the brain. Hence, it is discussed herein how the selected methodology influences the outcome and created knowledge of LAT1-utilizing compounds.

Pharmacological treatment of epilepsy is challenged by several barriers with the blood brain barrier (BBB) imposing the utmost restrictions to brain drug delivery. Antiepileptic drugs aim to reduce seizures frequency and severity while exerting minimal toxic effects. Herein, the merits of phytomedicine and brain targeted nanocarriers were combined for the control of seizures in a chronic epilepsy model. Fisetin (FS); a polyphenol and carveol (CAR); a limonene monoterpenoid were selected for their neuroprotective roles and co-loaded into lipid nanocapsules (FS/CAR@LNC). This was further decorated with mannose by post insertion targeting glucose transporter (GLUT-1) overexpressed on BBB (MAN-FS/CAR@LNC). The optimized MAN-FS/CAR@LNC revealed good colloidal properties with particle size (53.17 ± 4.06 nm), and low polydispersity index (0.11) and 26.9 ± 1.05 mV zeta potential. A high entrapment efficiency exceeding 99% and sustained drug release profile over 72 h were observed for both FS and CAR. In the in vivo imaging system (IVIS), MAN-FS/CAR@LNC attained 1.3-fold increase in fluorescence intensity at 5 h interval compared to FS/CAR@LNC. Following intraperitoneal administration in PTZ- induced chronic epilepsy mouse model, MAN-FS/CAR@LNC achieved maximal control of epileptic seizures accompanied with rehabilitation of locomotion, depressive and anxiety like-behaviors compared to FS/CAR and FS/CAR@LNC. Similarly, analysis of biomarkers reflecting depression and anxiety actions (brain-derived neurotrophic factor; BDNF, serotonin and glutamate) together with inflammatory markers (IL-6 and IL-1ꞵ) and histopathological assessment affirmed MAN-FS/CAR@LNC excelling in enhancing LNC brain targetability and hence seizures control. In conclusion, FS/CAR co-therapy aided with mannosylated LNC could present a compelling podium for the effective management of CNS disorders.