Application of 5-Aminolevulinic Acid and Its Derivatives for Photodynamic Therapy In Vitro and In Vivo

  • Asta Juzeniene
  • Petras Juzenas
  • Johan Moan
Part of the Methods in Molecular Biology book series (MIMB, volume 635)


Photodynamic therapy (PDT) with 5-aminolevulinic acid (ALA) is the most widely used form of PDT in clinical practice. Topical application of ALA leads to overproduction of the endogenous photosensitizer protoporphyrin IX (PpIX). ALA-PDT is efficient treatment of superficial skin lesions, but not for thicker lesions. The main reason for this is suboptimal penetration of ALA molecules through cellular membranes and through stratum corneum of intact skin. Different approaches (formulations, mechanical and physical penetration enhancers, ALA derivatives) are currently used to increase the penetration. The content and distribution of ALA intracellularly and in tissues is difficult to measure, but PpIX content, on a relative scale, can be easily measured by fluorimetric assays.

Key words

Photodynamic therapy 5-aminolevulinic acid 5-aminolevulinic acid derivatives protoporphyrin IX fluorescence spectroscopy extraction method 



We appreciate financial support from the Norwegian Cancer Society (Kreftforeningen). As well P. Juzenas would like to acknowledge researcher grant received from the Research Council of Norway (Forskningsrådet). The authors also thank Vladimir Iani and Dr. Li-Wei Ma for technical assistance, reading the manuscript and giving valuable comments and suggestions.


  1. 1.
    Peng, Q., Warloe, T., Berg, K., Moan, J., Kongshaug, M., Giercksky, K. E., and Nesland, J. M. (1997) 5-Aminolevulinic acid-based photodynamic therapy. Clinical research and future challenges. Cancer, 79, 2282–2308.PubMedCrossRefGoogle Scholar
  2. 2.
    Morton, C. A., McKenna, K. E., and Rhodes, L. E. (2008) Guidelines for topical photodynamic therapy: update. Br J Dermatol, 159, 1245–1266.PubMedCrossRefGoogle Scholar
  3. 3.
    Donnelly, R. F., McCarron, P. A., and Woolfson, A. D. (2005) Drug delivery of aminolevulinic acid from topical formulations intended for photodynamic therapy. Photochem Photobiol, 81, 750–767.PubMedCrossRefGoogle Scholar
  4. 4.
    Morrow, D. I., Garland, M. J., McCarron, P. A., Woolfson, A. D., and Donnelly, R. F. (2007) Innovative drug delivery strategies for topical photodynamic therapy using porphyrin precursors. J Environ Pathol Toxicol Oncol, 26, 105–116.PubMedCrossRefGoogle Scholar
  5. 5.
    Donnelly, R. F., McCarron, P. A., Morrow, D. I., Sibani, S. A., and Woolfson, A. D. (2008) Photosensitiser delivery for photodynamic therapy. Part 1: topical carrier platforms. Expert Opin Drug Deliv, 5, 757–766.PubMedCrossRefGoogle Scholar
  6. 6.
    Hua, Z., Gibson, S. L., Foster, T. H., and Hilf, R. (1995) Effectiveness of delta-aminolevulinic acid-induced protoporphyrin as a photosensitizer for photodynamic therapy in vivo. Cancer Res, 55, 1723–1731.PubMedGoogle Scholar
  7. 7.
    Luksiene, Z., Eggen, I., Moan, J., Nesland, J. M., and Peng, Q. (2001) Evaluation of protoporphyrin IX production, phototoxicity and cell death pathway induced by hexylester of 5-aminolevulinic acid in Reh and HPB-ALL cells. Cancer Lett, 169, 33–39.PubMedCrossRefGoogle Scholar
  8. 8.
    Sato, N., Moore, B. W., Keevey, S., Drazba, J. A., Hasan, T., and Maytin, E. V. (2007) Vitamin D enhances ALA-induced protoporphyrin IX production and photodynamic cell death in 3-D organotypic cultures of keratinocytes. J Invest Dermatol, 127, 925–934.PubMedCrossRefGoogle Scholar
  9. 9.
    Gaullier, J. M., Berg, K., Peng, Q., Anholt, H., Selbo, P. K., Ma, L. W., and Moan, J. (1997) Use of 5-aminolevulinic acid esters to improve photodynamic therapy on cells in culture. Cancer Res, 57, 1481–1486.PubMedGoogle Scholar
  10. 10.
    Moan, J., Ma, L. W., Juzeniene, A., Iani, V., Juzenas, P., Apricena, F., and Peng, Q. (2003) Pharmacology of protoporphyrin IX in nude mice after application of ALA and ALA esters. Int J Cancer, 103, 132–135.PubMedCrossRefGoogle Scholar
  11. 11.
    Margalit, R., Shaklai, N., and Cohen, S. (1983) Fluorimetric studies on the dimerization equilibrium of protoporphyrin IX and its haemato derivative. Biochem J, 209, 547–552.PubMedGoogle Scholar
  12. 12.
    Moan, J. (1984) The photochemical yield of singlet oxygen from porphyrins in different states of aggregation. Photochem Photobiol, 39, 445–449.CrossRefGoogle Scholar
  13. 13.
    Juzenas, P., Juzeniene, A., Stakland, S., Iani, V., and Moan, J. (2002) Photosensitizing effect of protoporphyrin IX in pigmented melanoma of mice. Biochem Biophys Res Commun, 297, 468–472.PubMedCrossRefGoogle Scholar
  14. 14.
    Hanania, J. and Malik, Z. (1992) The effect of EDTA and serum on endogenous porphyrin accumulation and photodynamic sensitization of human K562 leukemic cells. Cancer Lett, 65, 127–131.PubMedCrossRefGoogle Scholar
  15. 15.
    Iinuma, S., Farshi, S. S., Ortel, B., and Hasan, T. (1994) A mechanistic study of cellular photodestruction with 5-aminolaevulinic acid-induced porphyrin. Br J Cancer, 70, 21–28.PubMedCrossRefGoogle Scholar
  16. 16.
    Gomer, C. J., Jester, J. V., Razum, N. J., Szirth, B. C., and Murphree, A. L. (1985) Photodynamic therapy of intraocular tumors: examination of hematoporphyrin derivative distribution and long-term damage in rabbit ocular tissue. Cancer Res, 45, 3718–3725.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Asta Juzeniene
    • 1
  • Petras Juzenas
    • 1
  • Johan Moan
    • 1
    • 2
  1. 1.Department of Radiation BiologyInstitute for Cancer Research, The Norwegian Radium Hospital, MontebelloOsloNorway
  2. 2.Institute of PhysicsUniversity of OsloOsloNorway

Personalised recommendations