Chemical Reactivity Window Determines Prodrug Efficiency toward Glutathione Transferase Overexpressing Cancer Cells

• Published in: Molecular pharmaceutics Vol. 13; no. 6; pp. 2010 - 2025
• Main Authors: van Gisbergen, Marike W, Cebula, Marcus, Zhang, Jie, Ottosson-Wadlund, Astrid, Dubois, Ludwig, Lambin, Philippe, Tew, Kenneth D, Townsend, Danyelle M, Haenen, Guido R. M. M, Drittij-Reijnders, Marie-José, Saneyoshi, Hisao, Araki, Mika, Shishido, Yuko, Ito, Yoshihiro, Arnér, Elias S. J, Abe, Hiroshi, Morgenstern, Ralf, Johansson, Katarina
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 06.06.2016
• Subjects: Antineoplastic Agents - pharmacology; Cell Line, Tumor; Cytostatic Agents - pharmacology; Doxorubicin - pharmacology; Drug Resistance, Neoplasm - drug effects; Etoposide - pharmacology; Glutathione - metabolism; Glutathione Transferase - metabolism; Humans; MCF-7 Cells; Medicin och hälsovetenskap; Prodrugs - pharmacology; Sulfonamides - pharmacology; Up-Regulation - drug effects; glutathione transferases; GSTA1; MGST1; etoposide; doxorubicin; prodrugs; TrxR1; redox
• ISSN: 1543-8384; 1543-8392; 1543-8392
• DOI: 10.1021/acs.molpharmaceut.6b00140

Glutathione transferases (GSTs) are often overexpressed in tumors and frequently correlated to bad prognosis and resistance against a number of different anticancer drugs. To selectively target these cells and to overcome this resistance we previously have developed prodrugs that are derivatives of existing anticancer drugs (e.g., doxorubicin) incorporating a sulfonamide moiety. When cleaved by GSTs, the prodrug releases the cytostatic moiety predominantly in GST overexpressing cells, thus sparing normal cells with moderate enzyme levels. By modifying the sulfonamide it is possible to control the rate of drug release and specifically target different GSTs. Here we show that the newly synthesized compounds, 4-acetyl-2-nitro-benzenesulfonyl etoposide (ANS–etoposide) and 4-acetyl-2-nitro-benzenesulfonyl doxorubicin (ANS–DOX), function as prodrugs for GSTA1 and MGST1 overexpressing cell lines. ANS–DOX, in particular, showed a desirable cytotoxic profile by inducing toxicity and DNA damage in a GST-dependent manner compared to control cells. Its moderate conversion of 500 nmol/min/mg, as catalyzed by GSTA1, seems hereby essential since the more reactive 2,4-dinitrobenzenesulfonyl doxorubicin (DNS–DOX) (14000 nmol/min/mg) did not display a preference for GSTA1 overexpressing cells. DNS–DOX, however, effectively killed GSTP1 (20 nmol/min/mg) and MGST1 (450 nmol/min/mg) overexpressing cells as did the less reactive 4-mononitrobenzenesulfonyl doxorubicin (MNS–DOX) in a MGST1-dependent manner (1.5 nmol/min/mg) as shown previously. Furthermore, we show that the mechanism of these prodrugs involves a reduction in GSH levels as well as inhibition of the redox regulatory enzyme thioredoxin reductase 1 (TrxR1) by virtue of their electrophilic sulfonamide moiety. TrxR1 is upregulated in many tumors and associated with resistance to chemotherapy and poor patient prognosis. Additionally, the prodrugs potentially acted as a general shuttle system for DOX, by overcoming resistance mechanisms in cells. Here we propose that GST-dependent prodrugs require a conversion rate “window” in order to selectively target GST overexpressing cells, while limiting their effects on normal cells. Prodrugs are furthermore a suitable system to specifically target GSTs and to overcome various drug resistance mechanisms that apply to the parental drug.