title: L-Type amino acid transporter 1 (lat1)-mediated targeted delivery of perforin inhibitors.
abstract: Perforin is a cytolytic pore-forming glycoprotein secreted by cytotoxic effector cells. It is a key component of the immune response against virus-infected and transformed cells and has been implicated in a number of human diseases. Perforin activity can be inhibited by small-molecular-weight compounds, although less is known about their delivery to the site of action. Therefore, in the present study, it was explored if perforin inhibitors could be efficiently and site-selectively delivered firstly into the cytotoxic effector cells and secondly into lytic granules, in which perforin is stored. This was accomplished by designing and synthesizing four prodrugs of perforin inhibitors that could utilize l-type amino acid transporter (LAT1), since activated immune cells are known to over-express LAT1. The results demonstrate that cellular uptake of perforin inhibitors can be increased by LAT1-utilizing prodrugs (into human breast adenocarcinoma cells (MCF-7)). Furthermore, these prodrugs were also able to deliver perforin inhibitors into the cell organelles having lower pH (rat liver lysosomes). Therefore, by using these prodrugs, intracellular mechanisms of perforin inhibitory activity can be studied more thoroughly in future. Moreover, this prodrug approach can be applied for other drugs that would benefit from targeted delivery into cells expressing LAT1, such as cancer.
authors.affiliation: School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, P.O. Box 1627, FI-70211 Kuopio, Finland. Electronic address: Kristiina.Huttunen@uef.fi.
authors.forename: Kristiina M
authors.initials: KM
authors.lastname: Huttunen
authors.affiliation: School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, P.O. Box 1627, FI-70211 Kuopio, Finland.
authors.forename: Johanna
authors.initials: J
authors.lastname: Huttunen
authors.affiliation: School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, P.O. Box 1627, FI-70211 Kuopio, Finland.
authors.forename: Imke
authors.initials: I
authors.lastname: Aufderhaar
authors.affiliation: School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, P.O. Box 1627, FI-70211 Kuopio, Finland.
authors.forename: Mikko
authors.initials: M
authors.lastname: Gynther
authors.affiliation: Auckland Cancer Society Research Centre, The University of Auckland, Private Bag 92019, Auckland 1142, New Zealand.
authors.forename: William A
authors.initials: WA
authors.lastname: Denny
authors.affiliation: Auckland Cancer Society Research Centre, The University of Auckland, Private Bag 92019, Auckland 1142, New Zealand.
authors.forename: Julie A
authors.initials: JA
authors.lastname: Spicer
mesh_terms: D000818:Animals; D004305:Dose-Response Relationship, Drug; D016503:Drug Delivery Systems / Q000379:methods*; D006801:Humans; D027283:Large Neutral Amino Acid-Transporter 1 / Q000008:administration & dosage* / Q000737:chemistry; D061986:MCF-7 Cells; D054353:Perforin / Q000037:antagonists & inhibitors*; D011355:Prodrugs / Q000008:administration & dosage* / Q000737:chemistry; D051381:Rats
chemical_list: D027283:Large Neutral Amino Acid-Transporter 1; D011355:Prodrugs; D054353:Perforin
keywords: Perforin inhibitor; Prodrug; Targeted drug delivery; Transporter-mediated drug delivery; l-type amino acid Transporter 1 (LAT1)