The alkylating prodrug J1 can be activated by aminopeptidase N, leading to a possible target directed release of melphalan

Malin Wickström; Kristina Viktorsson; Lovisa Lundholm; Reidun Aesoy; Helen Nygren; Linda Sooman; Mårten Fryknäs; Lotte Vogel; Rolf Lewensohn; Rolf Larsson; Joachim Gullbo

doi:10.1016/j.bcp.2009.12.022

The alkylating prodrug J1 can be activated by aminopeptidase N, leading to a possible target directed release of melphalan

Malin Wickström, Kristina Viktorsson, Lovisa Lundholm, Reidun Aesoy, Helen Nygren, Linda Sooman, Mårten Fryknäs, Lotte Vogel, Rolf Lewensohn, Rolf Larsson, Joachim Gullbo

Institut for Cellulær og Molekylær Medicin

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

53 Citationer (Scopus)

Abstract

Udgivelsesdato: 2010-May-1

Originalsprog	Engelsk
Tidsskrift	Biochemical Pharmacology
Vol/bind	79
Udgave nummer	9
Sider (fra-til)	1281-1290
Antal sider	9
ISSN	0006-2952
DOI	https://doi.org/10.1016/j.bcp.2009.12.022
Status	Udgivet - 2010

Bibliografisk note

Adgang til dokumentet

10.1016/j.bcp.2009.12.022

Citationsformater

@article{b7fd89d02d0f11df8ed1000ea68e967b,

title = "The alkylating prodrug J1 can be activated by aminopeptidase N, leading to a possible target directed release of melphalan",

abstract = "The alkylating prodrug of melphalan, J1 (melphalanyl-l-p-fluorophenylalanyl ethyl ester) is currently in early clinical trials. Preclinical studies have shown that J1-mediated cytotoxicity is dependent on hydrolytic activity of tumor cells. In this report we have analyzed potential peptidases and esterases of importance for release of free melphalan from J1. Exposure of tumor cell lines to J1 resulted in a significant increased level of free intracellular melphalan, at least tenfold at C(max), compared to exposure to melphalan at the same molar concentration. This efficient intracellular delivery could be inhibited in both magnitude and in time by bestatin, a broad spectrum inhibitor of the aminopeptidases, including the metalloproteinase aminopeptidase N (APN, EC 3.4.11.2.), and ebelactone A, an esterase inhibitor. These effects resulted, as expected, in decreased cytotoxic effects of J1. A specific role of APN in hydrolyzing J1 releasing free melphalan was demonstrated in vitro with pure APN enzyme. By using plasmid-based overexpression of APN or down regulation of endogenous APN with siRNA in different tumor cell lines we here confirm the involvement of APN in J1-mediated cytotoxic and apoptotic signaling. In conclusion, this study demonstrates a role of APN in the activation of the melphalan prodrug J1 and subsequently, its cytotoxicity. Given that APN is shown to be overexpressed in several solid tumors our data suggest that J1 may be activated in a tumor selective manner.",

author = "Malin Wickstr{\"o}m and Kristina Viktorsson and Lovisa Lundholm and Reidun Aesoy and Helen Nygren and Linda Sooman and M{\aa}rten Frykn{\"a}s and Lotte Vogel and Rolf Lewensohn and Rolf Larsson and Joachim Gullbo",

year = "2010",

doi = "10.1016/j.bcp.2009.12.022",

language = "English",

volume = "79",

pages = "1281--1290",

journal = "Biochemical Pharmacology",

issn = "0006-2952",

publisher = "Elsevier",

number = "9",

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TY - JOUR

T1 - The alkylating prodrug J1 can be activated by aminopeptidase N, leading to a possible target directed release of melphalan

AU - Wickström, Malin

AU - Viktorsson, Kristina

AU - Lundholm, Lovisa

AU - Aesoy, Reidun

AU - Nygren, Helen

AU - Sooman, Linda

AU - Fryknäs, Mårten

AU - Vogel, Lotte

AU - Lewensohn, Rolf

AU - Larsson, Rolf

AU - Gullbo, Joachim

PY - 2010

Y1 - 2010

N2 - The alkylating prodrug of melphalan, J1 (melphalanyl-l-p-fluorophenylalanyl ethyl ester) is currently in early clinical trials. Preclinical studies have shown that J1-mediated cytotoxicity is dependent on hydrolytic activity of tumor cells. In this report we have analyzed potential peptidases and esterases of importance for release of free melphalan from J1. Exposure of tumor cell lines to J1 resulted in a significant increased level of free intracellular melphalan, at least tenfold at C(max), compared to exposure to melphalan at the same molar concentration. This efficient intracellular delivery could be inhibited in both magnitude and in time by bestatin, a broad spectrum inhibitor of the aminopeptidases, including the metalloproteinase aminopeptidase N (APN, EC 3.4.11.2.), and ebelactone A, an esterase inhibitor. These effects resulted, as expected, in decreased cytotoxic effects of J1. A specific role of APN in hydrolyzing J1 releasing free melphalan was demonstrated in vitro with pure APN enzyme. By using plasmid-based overexpression of APN or down regulation of endogenous APN with siRNA in different tumor cell lines we here confirm the involvement of APN in J1-mediated cytotoxic and apoptotic signaling. In conclusion, this study demonstrates a role of APN in the activation of the melphalan prodrug J1 and subsequently, its cytotoxicity. Given that APN is shown to be overexpressed in several solid tumors our data suggest that J1 may be activated in a tumor selective manner.

AB - The alkylating prodrug of melphalan, J1 (melphalanyl-l-p-fluorophenylalanyl ethyl ester) is currently in early clinical trials. Preclinical studies have shown that J1-mediated cytotoxicity is dependent on hydrolytic activity of tumor cells. In this report we have analyzed potential peptidases and esterases of importance for release of free melphalan from J1. Exposure of tumor cell lines to J1 resulted in a significant increased level of free intracellular melphalan, at least tenfold at C(max), compared to exposure to melphalan at the same molar concentration. This efficient intracellular delivery could be inhibited in both magnitude and in time by bestatin, a broad spectrum inhibitor of the aminopeptidases, including the metalloproteinase aminopeptidase N (APN, EC 3.4.11.2.), and ebelactone A, an esterase inhibitor. These effects resulted, as expected, in decreased cytotoxic effects of J1. A specific role of APN in hydrolyzing J1 releasing free melphalan was demonstrated in vitro with pure APN enzyme. By using plasmid-based overexpression of APN or down regulation of endogenous APN with siRNA in different tumor cell lines we here confirm the involvement of APN in J1-mediated cytotoxic and apoptotic signaling. In conclusion, this study demonstrates a role of APN in the activation of the melphalan prodrug J1 and subsequently, its cytotoxicity. Given that APN is shown to be overexpressed in several solid tumors our data suggest that J1 may be activated in a tumor selective manner.

U2 - 10.1016/j.bcp.2009.12.022

DO - 10.1016/j.bcp.2009.12.022

M3 - Journal article

C2 - 20067771

SN - 0006-2952

VL - 79

SP - 1281

EP - 1290

JO - Biochemical Pharmacology

JF - Biochemical Pharmacology

IS - 9

ER -