Talanta 89 (2012) 91–98 Contents lists available at SciVerse ScienceDirect Talanta j ourna l ho me page: www.elsevier.com/locate/talanta Metabonomic study on the cumulative cardiotoxicity of a pirarubicin liposome powder Wenjuan Cong a , Qionglin Liang b , Li Li c , Jun Shi d , Qingfei Liu e , Yi Feng a , Yiming Wang b , Guoan Luo b,∗ a Engineering Research Center of Modern Preparation Technology of TCM, Ministry of Education, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, PR China b Chemistry Department, Tsinghua University, Beijing 100084, PR China c The Center Laboratory, Post-doctoral Research Center, The Second College of Clinical Medicine, Guangzhou University of Chinese Medicine, Guangzhou 510120, PR China d School of Pharmacy, East China University of Science and Technology, Shanghai 200237, PR China e School of Medicine, Tsinghua University, Beijing 100084, PR China a r t i c l e i n f o Article history: Received 20 August 2011 Received in revised form 18 November 2011 Accepted 22 November 2011 Available online 29 November 2011 Keywords: Pirarubicin liposome powders Cumulative cardiotoxicity Molecular mechanism Metabonomics In vitro cytotoxicity a b s t r a c t Pirarubicin (THP) is an anthracycline frequently used in the chemotherapy against acute leukemia, malig- nant lymphoma and several solid tumors. However, its clinical use is severely limited by the development of a progressive dose-dependent cardiomyopathy that results in irreversible congestive heart failure. To provide a strategy for constraining or minimizing the cumulative cardiotoxicity of THP, a pirarubicin liposome powder (L-THP) was appropriately prepared, and the cumulative cardiotoxicity of L-THP and free THP (F-THP) were investigated on Sprague-Dawley rats after 3 successive doses. Urinary samples for metabonomic study, serum samples for biochemical assay, and heart samples for histopathology test were collected. As a result, the metabonomics-based findings such as PLS-DA plotting showed minimal metabolic alterations in L-THP as compared to F-THP, and correlated with the changes of serum biochem- ical assay and cardiac histopathology as measurements of damage to heart tissue. Our results confirm that when encapsulated into liposomes, the cumulative cardiotoxicity of THP can be greatly ameliorated. Lipophilic aglycone metabolites of THP associated with redox cycling are cardiotoxic for the possibility of reactive oxygen species (ROS) formation. Also, metabonomic analysis shows that the successive doses of THP will lead to severe metabolic pathways disturbances in the cell energy production. Further, the preliminary efficacy study of L-THP on lung cancer was evaluated in the approach of in vitro cytotoxicity on A549 cells by high content screening (HCS) analysis, and L-THP was found to exhibit better therapeutic index against lung cancer than THP. Crown Copyright © 2011 Published by Elsevier B.V. All rights reserved. 1. Introduction Anthracyclines, notably doxorubicin (DOX), epirubicin and daunorubicin, are potent antineoplastic agents used extensively to treat a range of cancers, including leukemias, lymphomas, sarcomas and carcinomas [1–4]. However, the clinical use of anthracy- clines can be reviewed as a sort of double-edged sword. On the one hand, they can play an undisputed key role in the cancer therapy; on the other hand, their successive doses will induce irre- versible cardiomyopathy that can be lethal even after the cessation of treatment [5]. Pirarubicin (THP) (Fig. 1), a-tetrahydropyranyl (DOX), is often used in the chemotherapy against acute leukemia, malignant lymphoma and several solid tumors. Although THP has been frequently reported to exhibit satisfactory cardiotoxicity at a single dose in the anthracycline family [6,7], there is still a risk ∗ Corresponding author at: Department of Chemistry, Tsinghua University, Haid- ian District, Beijing 100084, PR China. Tel.: +86 10 62781688; fax: +86 10 62781688. E-mail address: luoga@tsinghua.edu.cn (G. Luo). of cardiomyopathy at cumulative doses. For instance, THP causes a significant decrease in the left ventricular ejection fraction at a cumulative dose of 460 mg/m 2 or can lead to full-blown conges- tive heart failure (CHF) at greater than 500 mg/m 2 for women with metastatic breast cancer [8]. Also, THP can cause severe cardiac dysfunction at a cumulative dose of 360 mg/m 2 for elderly patients with non-Hodgkin’s lymphoma [9]. In addition, dilative cardiomy- opathy and CHF can develop after the completion of cumulative THP regiments [5]. Therefore, there is a need to identify ways to prevent or alleviate the cumulative cardiotoxicity of THP in order to improve its therapeutic index. A liposomal drug delivery system, a proven strategy for constraining or preventing anthracycline-induced cardiotoxicity [10,11], can alter the in vivo behavior, alleviate the severe side effects and thus improving the therapeutic index of the encap- sulated drug [12,13]. Pharmacokinetic studies have shown that the heart accumulation of THP can be greatly reduced after a sin- gle dose when encapsulated into liposomes [14,15]. However, no studies to date are concerned to the assessment of cumulative cardiotoxicity of pirarubicin liposomes (L-THP). In addition, the 0039-9140/$ – see front matter. Crown Copyright © 2011 Published by Elsevier B.V. All rights reserved. doi:10.1016/j.talanta.2011.11.071