Example of a Bullet Point Slide Medical faculty Department of pharmacy University Novi Sad Bullet Point 1 Bullet Point 2 Sub Bullet PROTECTIVE EFFECT OF FULLERENOL C60(OH)24 ON LIPID PEROXIDATION OF RAT TISSUES TREATED WITH DOXORUBICIN Vukosava Djordjevic Milic1, Branislava Srdjenovic1, Nevena Grujic1, Aleksandar Djordjevic2, Viktorija Dragojevic Simic3, Velibor Vasovic4 1Department of Pharmacy, Faculty of Medicine, University of Novi Sad, Serbia 2 Department of Chemistry, Faculty of Sciences, University of Novi Sad, Serbia 3Centre for Clinical Pharmacology, Military Medical Academy, Belgrade, Serbia 4Department of Pharmacology , Faculty of Medicine, University of Novi Sad, Serbia

The anthracycline antibiotic doxorubicin (DOX) is among the most important antitumor agents 14-hydroksy-daunomycin Doxorubicin is used for more than 30 years in treatment of variety of malignancies. breast cancer, tumors of bile duct, endometrial tissue, the esophagus and liver, osteosarcomas, soft tissue sarcomas and non-Hodgkin’s lymphoma adverse effects such as myelosuppression and cardiotoxicity, limits the use of doxorubicin.

Number of different mechanisms has been proposed for the cytostatic and cytotoxic activities: intercalation into DNA with consequent inhibition of macromolecular biosynthesis, free radical formation with consequent induction of DNA damage or lipid peroxidation, direct membrane effects and the initiation of DNA damage through inhibition of topoisomerase II in the interaction of these drugs with tumor cell DNA binding and alkylation, DNA cross-linking, Among the mechanisms of doxorubicin toxicity, the most plausible seems to be the increase of free radical production, which induces lipid peroxidation and oxidative damage of cells.

Doxorubicin free radical formation 7-deoxyaglycon

Lipid peroxidation Alteration of enyzme funiton, Formation of O2•− modulation of sinthesys of DNA, transcription of RNA Induction of lipid peroxidation. Formation of O2•− Polyunsturated faty acids within phospholipids bilayer of membrane are especially vulnerable to oxidative injury caused by ROS. end product of lipid peroxidation is malonyldialdehid malondialdehyde (MDA).

N-acetil cystein + vitamin E Probucol Deksrazoxan (ICRF -187) Toxic effects of doxorubicin could be decreased by: Changing the dosage schedule Deveoping of new anthracycline antibiotics which are not cardiotoxic Usage of cytoprotectors Glutathion N-acetil cystein + vitamin E Probucol Deksrazoxan (ICRF -187) Amifostin use of antioxidants as protectors against cell damage by oxidative injury could be potential solution to reduction of toxicity induced by doxorubicin.

(Harold W. Kroto, Richard E. Smalley i Robert E. Curl) fullerenes Fullerenes are the third allotropic modification of carbon, disocovered in 1985. (Harold W. Kroto, Richard E. Smalley i Robert E. Curl)

fundamental properties of fullerenes such as: extremely high hydrophobicity photo activity high cohesive force between fullerene molecules ability to accept and release electrons and relatively high reactivity allow their structural modification combinations of nucleophilic and electrophilic additions, cycloadditions and radical additions it is possible to covalently bond any class of organic compounds to a fullerene core

Fullerenol C60(OH)24 Previous investigation discover that in nanomole concentration and without photoinduction fullerenol shows : Radioprotectivity Antioxidant activity in in vitro systems Cytotoxicity against some tumor cell lines It may function as a free radical scavenger and it strongly suppresses cytotoxicity of doxorubicin in the model system.

The aim of this study was to investigate the toxic effect of doxorubicin on the rat experimental model and to test usage of a potential protector fullerenol in combination with doxorubicin by measuring product of lipid peroxidation in heart, liver, kidneys, testis and lungs.

Chemicals Doksorubicin hydrochloride Comercial, for i.v. applicaiton - Adrablastina®, Pharmacia & Upjohn, Italy. Fullerenol C60(OH)24 was synthesized by original method on the Depratment of Chemitrstry, Faculty of sciences, University Novi Sad

Experimental animal model Wistar male rats, wight 180 to 220g, old 8 to 10 weeks Standard laboratory conditions (room temperature, air humidity, light/dark cycle 12/12, water and food ad libidum I control group (0.9% saline i.p.) II doksorubicin (10 mg/ml i.p.) III fullerenol 50 mg/kg i.p. 30 minuts before doksorubicin (10 mg/ml i.p.) IV fullerenol 100 mg/kg i.p. 30 minuts before doksorubicin (10 mg/ml i.p.) V fullerenol 100 mg/kg i.p Second and fourteenth day after the treatment, animals were anesthetisized and organs for further analysis were taken. The samples were homogenized and used for spectrophotometric measurement of the products of lipid peroxidation

___________________________________________________Results______ heart *      liver       *p<0.05 related to I; p<0.05 related to II; p<0.05 realted to 2. day

___________________________________________________Results______       *p<0.05 realted to I;  p<0.05 related to II

___________________________________________________Results______ *p<0.05 realted to I;  p<0.05 related to II

Conclusion Results showed that 2. and 14. days after the treatment, lipid peroxidation was significantly increased only in group treated with doxorubicin Fullerenol, given as a pretreatment, in both applied dosage, sustained value of the lipid peroxidation of all examined tissues in the level of control Significantly higher levels of lipid peroxidation, of those obtained in control group, were noticed only in lungs . Dosage of fullerenol of 100 mg/kg showed better protective effects. Lipid peroxidation in organs of animals treated only with fullerenol in dosage of 100 mg/kg, was in the control level These finding imply that fullerenol protect examined organs from oxidative injury caused by doxorubicin, and exhibits antioxidative properties.

This work is part of the scientific project funded by Ministry of Science of Republic of Serbia grant No 142076