Formation of protoporphyrin IX from carboxylic- and amino-derivatives of 5-aminolevulinic acid

Summary 

Background:

Stability of ALA is an important factor for photodynamic therapy (PDT). The dimerization of ALA to pyrazines takes place via the amine group. It is, therefore, to be expected that blocking this group by addition of a formyl group should result in a more stable compound.

Methods:

The ability of a new N-formyl derivative of ALA (N-f-ALA) to form protoporphyrin IX (PPIX) was compared with that of ALA and three of its ester (methyl, butyl and hexyl) derivatives. Dark toxicity of the compounds was measured using MTT assay. Formation of PPIX was measured by fluorescence spectroscopy.

Results and conclusions:

N-f-ALA showed an outstanding stability in water solutions even at pH 7. However, it induced no PPIX neither in WiDr cells in vitro, nor in mouse skin in vivo. A probable reason is lack of an enzyme that can cleave the bond between the formyl group and ALA. Thus, steric hindrance may prevent processing of the compound into porphobilinogen. N-f-ALA did not inhibit PpIX formation from ALA and is unable to enter the heme cycle. Generation of ALA from its derivatives, therefore, seems to be an essential step in PPIX synthesis.

Keywords: Photodynamic therapy, Aminolevulinic acid derivatives, Stability, Carcinoma cell line