Safe and targeted anticancer efficacy of a novel class of antioxidant-conjugated difluorodiarylidenyl piperidones: Differential cytotoxicity in healthy and cancer cells

doi:10.1016/j.freeradbiomed.2010.02.009

Free Radical Biology and Medicine

Volume 48, Issue 9, 1 May 2010, Pages 1228-1235

https://doi.org/10.1016/j.freeradbiomed.2010.02.009 Get rights and content

Abstract

The development of smart anticancer drugs that can selectively kill cancer cells while sparing the surrounding healthy tissues/cells is of paramount importance for safe and effective cancer therapy. We report a novel class of bifunctional compounds based on diarylidenyl piperidone (DAP) conjugated to an N-hydroxypyrroline (NOH; a nitroxide precursor) group. We hypothesized that the DAP would have cytotoxic (anticancer) activity, whereas the NOH moiety would function as a tissue-specific modulator (antioxidant) of cytotoxicity. The study used four DAPs, namely H-4073 and H-4318 without NOH and HO-3867 and HO-4200 with NOH substitution. The goal of the study was to evaluate the proof-of-concept anticancer-versus-antioxidant efficacy of the DAPs using a number of cancerous (breast, colon, head and neck, liver, lung, ovarian, and prostate cancer) and noncancerous (smooth muscle, aortic endothelial, and ovarian surface epithelial) human cell lines. Cytotoxicity was determined using an MTT-based cell viability assay. All four compounds induced significant loss of cell viability in cancer cells, whereas HO-3867 and HO-4200 showed significantly less cytotoxicity in noncancerous cells. EPR measurements showed a metabolic conversion of the N-hydroxylamine function to nitroxide with significantly higher levels of the metabolite and superoxide radical-scavenging (antioxidant) activity in noncancerous cells compared to cancer cells. Western blot analysis showed that the DAP-induced growth arrest and apoptosis in cancer cells were mediated by inhibition of STAT3 phosphorylation at the Tyr705 and Ser727 residues and induction of apoptotic markers of cleaved caspase-3 and PARP. The results suggest that the antioxidant-conjugated DAPs will be useful as safe and effective anticancer agents for cancer therapy.

Section snippets

Chemicals

Superoxide dismutase (SOD), 6-carboxy-2′,7′-dichlorodihydrofluorescein diacetate, diacetoxymethyl ester (H₂DCF-DA), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and antibodies against actin were obtained from Sigma (St. Louis, MO, USA). 5-Diethoxyphosphoryl-5-methyl-1-pyrroline-N-oxide (DEPMPO) was from Radical Vision (Jerome-Marseille, France). Cell culture medium (RPMI 1640), fetal bovine serum (FBS), antibiotics, sodium pyruvate, trypsin, and phosphate-buffered saline

Cytotoxicity of DAPs to cancer cells

The cytotoxicity of DAPs (H-4073, HO-3867, H-4318, HO-4200) to established human cancer cell lines, namely A27820, A2780R, MCF-7, HCT-116, PC-3, HepG2, A549, and SCC4, was evaluated by exposing the cells to 10 μM concentration of the compounds for 24 h. All four compounds induced a substantial loss of cell viability in all the human cancer cell lines tested (Fig. 1). In particular, H-4073 and H-4318 exhibited higher toxicity compared to HO-3867 or HO-4200. The results further indicated that the

Discussion

The development of smart anticancer agents that selectively destroy cancer cells while sparing the surrounding healthy tissues/cells is the main goal of cancer therapy. The results of this study show that all four DAPs induce potential cytotoxicity in cancer cells. The N-hydroxypyrroline-conjugated DAPs, HO-3867 and HO-4200, although equally toxic to cancer cells, are significantly less toxic to noncancerous (healthy) cells. The differential cytotoxicity is mediated through inhibition of STAT3

Acknowledgments

This work was supported by National Institutes of Health Grant CA102264 (P.K.), a Kaleidoscope of Hope Foundation grant (K.S.), and Hungarian Research Fund Grant OTKA K81123 (K.H.).

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Original ContributionSafe and targeted anticancer efficacy of a novel class of antioxidant-conjugated difluorodiarylidenyl piperidones: Differential cytotoxicity in healthy and cancer cells

Abstract

Section snippets

Chemicals

Cytotoxicity of DAPs to cancer cells

Discussion

Acknowledgments

Blood

Bioorg. Med. Chem.

J. Biol. Chem.

Exp. Toxicol. Pathol.

J. Biol. Chem.

J. Biol. Chem.

Cell

Free Radic. Biol. Med.

Free Radic. Biol. Med.

Blood

Blood

Neoplasia

Curcumin suppresses Janus kinase–STAT inflammatory signaling through activation of Src homology 2 domain-containing tyrosine phosphatase 2 in brain microglia

J. Immunol.

Curcumin induces proapoptotic effects against human melanoma cells and modulates the cellular response to immunotherapeutic cytokines

Mol. Cancer Ther.

Curcumin induces G2/M arrest and apoptosis in cisplatin-resistant human ovarian cancer cells by modulating Akt and p38 MAPK

Cancer Biol. Ther.

Curcumin inhibits tumor growth and angiogenesis in ovarian carcinoma by targeting the nuclear factor-kappaB pathway

Clin. Cancer Res.

Curcumin induces apoptosis-independent death in oesophageal cancer cells

Br. J. Cancer

Curcumin inhibits Akt/mammalian target of rapamycin signaling through protein phosphatase-dependent mechanism

Mol. Cancer Ther.

Inhibition of vascular smooth-muscle cell proliferation and arterial restenosis by HO-3867, a novel synthetic curcuminoid, through up-regulation of PTEN expression

J. Pharmacol. Exp. Ther

EF24, a novel synthetic curcumin analog, induces apoptosis in cancer cells via a redox-dependent mechanism

Anticancer Drugs

Diphenyl difluoroketone: a curcumin derivative with potent in vivo anticancer activity

Cancer Res.

Evaluation of a novel class of fluorinated curcumin analogs for safe and targeted anticancer therapy (STAT)

Free Radic. Biol. Med.

Recent advances in protection against doxorubicin-induced toxicity

Technol. Cancer Res. Treat.

Hydroxyl radical scavenger ameliorates cisplatin-induced nephrotoxicity by preventing oxidative stress, redox state unbalance, impairment of energetic metabolism and apoptosis in rat kidney mitochondria

Cancer Chemother. Pharmacol.

Dietary antioxidants during cancer chemotherapy: impact on chemotherapeutic effectiveness and development of side effects

Nutr. Cancer

Factors influencing nitroxide reduction and cytotoxicity in vitro

Antioxid. Redox Signal.

Protection against oxidative stress by nitroxides

Exp. Biol. Med. (Maywood)

Noninvasive imaging of tumor redox status and its modification by tissue glutathione levels

Cancer Res.

Original Contribution
Safe and targeted anticancer efficacy of a novel class of antioxidant-conjugated difluorodiarylidenyl piperidones: Differential cytotoxicity in healthy and cancer cells