Ketoprofen

Comparison of DNA Damage Photoinduced by Ketoprofen, Fenofibric Acid and Benzophenone via Electron and Energy Transfer(para)

ABSTRACT

Ketoprofen (KP) and fenofibrate, respectively, anti-inflammatory and hypolipidemiant agents, promote anormal photosensitivity in patients and may induce photoallergic cross-reactions correlated to their benzophenone-like structure. Here, their ability to photosensitize the degradation of biological targets was particularly investigated in DNA. The photosensitization of DNA damage by KP and fenofibric acid (FB), the main metabolite of fenofibrate, and their parent compound, benzophenone (BZ), was examined on a 32P-end-labeled synthetic oligonucleotide in phosphatebuffered solution using gel sequencing experiments. Upon irradiation at lambda > 320 nm, piperidine-sensitive lesions were induced in single-stranded oligonucleotides by KP, FB and BZ at all G sites to the same extent. This pattern of damage, enhanced in D^sub 2^O is characteristic of a Type-II mechanism. Spin trapping experiments using 2,2,6,6-tetramethyl-4-piperidone have confirmed the production of singlet oxygen during drug photolysis. On double-stranded oligonucleotides, highly specific DNA break occurred selectively at 5'-G of a 5'-GG-3' sequence, after alkali treatment. Prolonged irradiation led to the degradation of all G residues, with efficiency decreasing in the order 5'-GG > 51-GA > 51-GC > 5'-GT, in good agreement with the calculated lowest ionization potentials of stacked nucleobase models supporting the assumption of a Type-- I mechanism involving electron transfer, also observed to a lesser extent with adenine. Cytosine sites were also affected but the action of mannitol which selectively inhibited cytosine lesions suggests, in this case, the involvement of hydroxyl radical, also detected by electronic paramagnetic resonance using 5,5-dimethyl-1-- pyrrolidine-1-oxide as spin trap. On a double-stranded ^sup 32^P-end-labeled 25-mer oligonucleotide containing TT and TTT sequences, the three compounds were found to photosensitize by triplet-triplet energy transfer the

formation of cyclobutane thymine dimers detected using T4 endonuclease V.

Abbreviations: ATP, adenosine triphosphate; BZ, benzophenone; DMPO, 5,5-dimethyl-1-pyrrolidine-1-oxide; DMSO, dimethylsulfoxide; EDTA, ethylenediamine tetraacetic acid; EPR, electronic paramagnetic resonance; FB, fenofibric acid; 8-OxodAdo, 8-oxo-7,8-dihydro-2'-deoxyadenosine; KP, ketoprofen; NSAID, nonsteroidal antiinflammatory drug; SCE, saturated calomel electrode; SOD, superoxide dismutase; TMP, 2,2,6,6-tetramethyl-4-- piperidone.

CONCLUSIONS

Comparison of the photosensitizing activities of KP and FB to that of BZ on two double-stranded oligonucleotides, chosen to easily characterize the participation of electron and energy transfer, clearly shows that the three compounds have the similar ability to promote DNA damage by both processes. As shown by the sequence specificity of the alkali-labile sites, KP, FB and BZ promote transfer of an electron through a Type-I mechanism involving guanines and to a lesser extent adenines. They also cause damage to cytosines by a poorly efficient radical mechanism. When the oligonucleotide displays -TT- and -TTT- sites, the three compounds photosensitize the formation of cyclobutane thymine dimers via triplet-triplet energy transfer. The behavior of these compounds toward single-stranded oligonucleotides is different, as the involvement of singlet oxygen by a Type-II mechanism appears predominant. In all cases, the efficiency of the three compounds is nearly the same suggesting that the radicals formed during the photolysis of KP and FB are not significantly involved in the formation of alkali-labile sites. Owing to the fact that the DNA damage photosensitization occurs mainly through electron transfer, as is deduced from this work, the photosensitization efficiency depends on the excited states and not on the radical formation. The quenching of the excited states of BZ and its derivatives observed in the presence of nucleic acids and attributed either to electron or energy transfer could very well slow down their photodegradation. These conclusions point out that the photosensitizing properties of these benzophenone derivatives in DNA depend mainly on the benzophenone chromophore. The problem is more complex in a biological medium, and these conclusions have not yet been extended directly to other biological targets or cells.

The ability of these compounds to photoinduce the formation of cyclobutane thymine dimers, which are at the origin of many biological lesions, is now clearly evidenced by this work. It should be noted that it is the first time that this reaction is shown with drugs, and furthermore, with commercial phototoxic drugs.

Acknowledgement-The authors thank Dr. De Montauzon for his technical assistance in electrochemistry.

(para)Posted on the website on 17 August 2001.

REFERENCES

1. Cusano, F., R. Rafenelli, R. Bacchilega and G. Errico (1987) Photo-contact dermatitis from ketoprofen. Contact Dermatis 17, 108-109.

2. Mozzanica, N. and P. D. Piggatto (1990) Contact and photocontact allergy to ketoprofen, clinical and experimental study. Contact Dermatitis 23, 336-340.

3. Serrano, G., J. M. Fortea, J. M. Latasa, F. Millan, C. James, F. Bosca and M. A. Miranda (1992) Photosensitivity induced by fibric acid and derivatives and its relation to photocontact dermatitis to ketoprofen. J. Am. Acad. DermatoL 27, 204-208.

4. Leroy, D., A. Dompmartin, E. Loner, Y. Leport and C. Audebert (1990) Photosensitivity induced by fenofibrate. Photodermatol. Photoimmunol. Photomed. 7, 136-138.

5. Jeanmougin, M., A. Petit, J. R. Manciet, M. Sigal and L. Dubertret (1996) Eczema photoallergique de contact au ketoprofene. Ann. DermatoL Venerol. 123, 251-255.

6. Ljunggren, B. (1985) Propionic acid-derivated nonsteroidal antiinflammatory drugs are phototoxic in vitro. Photodermatology 2, 3-9.

7. Kochevar, I. E. (1989) Phototoxicity of non steroidal antiinflammatory drugs: coincidence or specific mechanism? Arch. Dermatol. 125, 824-826.

8. Lhiaubet, V., F. Gutierrez, F. Penaud-Berruyer, E. Amouyal, J. P. Daudey, R. Poteau, N. Chouini-Lalanne and N. Paillous (2000) Spectroscopic and theoretical studies of the excited states of fenofibric acid and ketoprofen in relation with their photosensitizing properties. New J. Chem. 24, 403-410.

9. Monti, S., S. Sortino, G. De Guidi and G. Marconi (1997) Photochemistry of 2-(3-benzoylphenyl)propionic acid (ketoprofen). Part 1. A picosecond and nanosecond time resolved study in aqueous solution. J. Chem. Soc., Faraday Trans. 93, 22692275.

10. Marguery, M. C., N. Chouini-Lalanne, J. C. Ader and N. Paillous (1998) Comparison of the DNA damage photoinduced by fenofibrate and ketoprofen, two phototoxic drugs of parent structure. Photochem. Photobiol. 68, 679-684.

11. Bosch, F. and M. A. Miranda (1998) Photosensitizing drugs containing the benzophenone chromophore. J. Photochem. Photobiol. B: Biol. 43, 1-26.

12. Bosch, F., M. A. Miranda, G. Carganico and D. Mauleon (1994) Photochemical and photobiological properties of ketoprofen associated with the benzophenone chromophore. Photochem. Photobiol. 60, 96-101.

13. Martinez, L. J. and J. C. Scaiano (1997) Transient intermediates in the laser flash photolysis of ketoprofen in aqueous solutions: unusual photochemistry for the benzophenone chromophore. J. Am. Chem. Soc. 119, 11066-11079.

14. Miranda, M. A., F. Bosch, F. Vargas and N. Canudas (1994) Photosensitization by fenofibrate. II. In vitro phototoxicity of the major metabolites. Photochem. Photobiol. 59, 171-174.

15. Costanzo, L. L., G. De Guidi, C. Condorelli, A. Cambria and M. Fama (1989) Molecular mechanism of drug photosensitization-II. Photohemolysis sensitized by ketoprofen. Photochem. Photobiol. 50, 359-365.

16. Bosch, F., G. Carganico, J. V. Castell, M. J. Gomez-Lechon, D. Hernandez, D. Mauleon, L. A. Martinez and M. A. Miranda (1995) Evaluation of ketoprofen (RS and RIS) phototoxicity by a battery of in vitro assays. J. Photochem. Photobiol. B: Biol. 31, 133-138.

17. Artuso, T., J. Bernadou, B. Meunier, J. Piette and N. Paillous (1991) Mechanism of DNA cleavage mediated by photoexcited

non-steroidal antiinflammatory drugs. Photochem. PhotobioL 54, 205-213.

18. Chouini-Lalanne, N., M. Defais and N. Paillous (1998) Nonsteroidal antiinflammatory drug-photosensitized formation of pyrimidine dimer in DNA. Biochem. Pharmacol. 55, 441-446.

19. Charlier, M., C. Helene and W. L. Carrier (1972) Photochemical reactions of aromatic ketones with nucleic acids and their components-Ill. Chain breakage and thymine dimerization in benzophenone photosensitized DNA. Photochem. Photobiol. 15, 527-536.

20. Greenstock, C. L. and H. E. Johns (1968) Photosensitized dimerization of pyrimidines. Bioch. Biophys. Res. Commun. 30, 21-27.

21. Wood, P. D. and R. W. Redmond (1996) Triplet state interactions between nucleic acid bases in solution at room temperature: intermolecular energy and electron transfer. J. Am. Chem. Soc. 118, 4256-4263.

22. Gut, 1. G., P. D. Wood and R. W. Redmond (1996) Interaction of triplet photosensitizers with nucleotides and DNA in aqueous solution at room temperature. J. Am. Chem. Soc. 118, 23662373.

23. Delatour, T., T. Douki, C. D'Ham and J. Cadet (1998) Photosensitization of thymine nucleobase by benzophenone through energy transfer, hydrogen abstraction and one-electron oxidation. J. Photochem. Photobiol. B: Biol. 44, 191-198.

24. Maxam, A. M. and W. Gilbert (1980) Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 65, 499-559.

25. Rubin, C. M. and C. W. Schmid (1980) Pyrimidine-specific chemical reactions useful for DNA sequencing. Nucleic Acids Res. 8, 4613-4619.

26. Saito, L, M. Takayama, H. Sugiyama and T. Nakamura (1996) Design of photoactivable DNA-cleaving amino-acids with high sequence selectivity, In DNA and RNA Cleavers and Chemotherapy of Cancer and Viral Diseases (Edited by B. Meunier), pp. 163-176. Kluwer Academic, Netherlands.

27. Sugiyama, H. and I. Saito (1996) Theorical studies of -GG-- . photocleavage of DNA via electron transfer: significant lowering of ionization potential and 5'-localization of HOMO of stacked GG bases in B-form DNA. J. Am. Chem. Soc. 118, 7063-7068.

28. Lommel, L. and P. C. Hanawalt (1991) Preparation and determination of T4 endonuclease V activity. Mutat. Res. 255, 183191.

29. Amankwa, L. and L. G. Chatten (1984) Electrochemical reduction of ketoprofen and its determination in pharmaceutical dosage forms by differential-pulse polarography. Analyst 109, 5760.

30. Nadjo, L. and J. M. Sav6ant (1971) Electrochemical reduction of substituted benzophenones and fluorenones in media of low proton availability.Mechanisms of the reductive cleavage of bromo and chlorobenzophenone. J. Electroanal. Chem. 30, 41-57.

31. Rehm, D. and A. Weller (1970) Kinetics of fluorescence quenching by electron and H-atom transfer. Isr. J. Chem. 8, 259-271.

32. Steenken, S. and S. V. Jovanovic (1997) How easily oxidizable is DNA? One electron reduction potentials of adenosine and guanosine radicals in aqueous solution. J. Am. Chem. Soc. 119, 617-618.

33. De la Pena, D., C. Marti, S. Nonell, L. A. Martinez and M. A. Martinez (1997) Time resolved near infrared studies on singlet oxygen production by the photosensitizing 2-aryl-propionic acids. Photochem. Photobiol. 65, 828-832.

34. Darmanyan, A. P. and C. S. Foote (1993) Solvent effects on singlet oxygen yield from n(pi)* and pi(pi)* triplet carbonyl compounds. J. Phys. Chem. 97, 5032-5035.

35. Castelli, F., G. De Guidi, S. Guiffrida, P. Miano and S. Sortino (1999) Molecular mechanism of photosensitization XIII: a combined differential scanning calorimetry and DNA photosensitization study in non steroidal antiinflammatory drugs-DNA interaction. Int. J. Pharmaceutics 184, 21-33.

36. Saito, I., M. Takayama, H. Sugiyama and K. Nakatani (1995) Photoinduced DNA cleavage via electron transfer: demonstra

tion that guanine residues located 5' to guanine are the most electron donating sites. J. Am. Chem. Soc. 117, 6406-6407.

37. Starrs, S. M. and R. J. H. Davies (2000) Sequence specificity of alkali-labile DNA damage photosensitized by suprofen. Photochem. Photobiol. 72, 291-297.

38. Nakatani, K., K. Fujisawa, C. Dohno, T. Nakamura and I. Saito (1998) p-Cyano substituted 5-benzoyldeoxyuridine as a novel electron accepting nucleobase for one electron oxidation of DNA. Tetrahedron Lett. 39, 5995-5998.

39. Adam, W., C. R. Saha-Moller and A. Schonberger (1997) Type I and type II photosensitized oxidative modification of 2'-deoxyguanosine (dGuo) by triplet excited ketones generated thermally from the 1,2-dioxetane HTMD. J. Am. Chem. Soc. 119, 719-723.

40. Douki, T. and J. Cadet (1999) Modification of DNA bases by photosensitized one-electron oxidation. Int. J. Radial Biol. 75, 571-581.

41. Burrows, C. J. and J. G. Muller (1998) Oxidative nucleobase modification leading to strand scission. Chem. Rev. 98, 11091151.

42. Radschuweit, A., H. H. Rittinger, P. Nuhn, W. Wohlrab and C. Huschka (2001) UV-induces formation of hydrogen peroxide based on the photochemistry of ketoprofen. Photochem. Photobiol. 73, 119-127.

43. Charlier, M. and C. Helene (1972) Photochemical reactions of

aromatic ketones with nucleic acids and their components-I. Purine and pyrimidine bases and nucleosides. Photochem. Photobiol. 15, 71-87.

44. Roth, H. D. and A. A. Lamola (1974) Chemically induced dynamic nuclear polarization and exchange broadening in an electron-transfer reaction. J. Am. Chem. Soc. 96, 6270-6275.

45. Peters, K. S. and J. Lee (1993) Picosecond dynamics of the photoreduction of benzophenone by DABCO. J. Phys. Chem. 97, 3761-3764.

46. Devadoss, C. and R. W. Fessenden (1990) Picosecond and nanosecond studies of the photoreduction of benzophenone by 1,4-diazabicyclo[2.2.21 octane: characterization of the transient. J. Phys. Chem. 94, 4540-4549.

47. Gentil, A., F. Le Page and A. Sarasin (1997) The consequence of translesional replication of unique UV-induced photoproducts. Biol. Chem. 378, 1287-1292.

48. Bourre, F., G. Renault, P. C. Seawell and A. Sarasin (1985) Distribution of ultraviolet-induced lesions in Simian Virus 40 DNA. Biochimie 67, 293-299.

49. Rahn, R. 0. (1979) Triplet sensitization of DNA. Acta Biol. Med. Germ. 38, 1225-1231.

50. Kochevar, I. E. and D. A. Dunn (1990) Photosensitized reactions of DNA: cleavage and addition, In Bioorganic Photochemistry, Photochemistry and the Nucleic Acids, Vol. 1 (Edited by H. Morrison), pp. 273-316. John Wiley and Sons, New York.

Virginie Lhiaubet, Nicole Paillous and Nadia Chouini-Lalanne*

Laboratoire des Interactions Moleculaires et Reactivite Chimique et Photochimique, Universite Paul Sabatier, Toulouse, France

Received 23 April 2001; accepted 9 August 2001

*To whom correspondence should be addressed at: Laboratoire des IMRCP, Universite Paul Sabatier, 118, route de Narbonne, Toulouse, France. Fax: 5-61-55-81-55; e-mail: lalanne@chimie. ups-tlse.fr

Copyright American Society of Photobiology Nov 2001
Provided by ProQuest Information and Learning Company. All rights Reserved