In vitro metabolism of 5-ALA esters derivatives in hairless mice skin homogenate and in vivo PpIX accumulation studies.
OBJECTIVE
In topical photodynamic therapy, 5-ALA and its esters are enzymatically converted in the endogenous photosensitizing compounds such as, for example, protoporphyrin IX (PpIX). In order to elucidate in more detail their enzymatic fate, we have determined in vitro the enzymatic degradation of methyl, butyl, hexyl, and octyl-5-ALA ester derivatives in skin homogenate. Furthermore, in vivo porphyrin accumulation was measured in healthy hairless mice skins.


METHODS
Hairless mouse skins were homogenized in isotonic phosphate buffer pH 7.4. 5-ALA esters were added, and aliquots were colleted for HPLC-fluorimetric determinations of remaining content of 5-ALA esters. Furthermore, oil-in-water emulsions containing esters were topically applied to mice skin for 6 h, and the amount of accumulated PpIX in the treated areas was determined by quantitative extraction and confocal fluorescence microscopy.


RESULTS
The enzymatic degradation of esters follows pseudo first-order kinetics. The octyl ester had the largest rate constant for enzymatic degradation, followed by hexyl-, butyl-, and methyl-ALA. The long-chained 5-ALA esters, butyl-, hexyl-, and octyl ester, induced significantly more porphyrins than 5-ALA and 5-ALA methyl ester as shown by confocal microscopy and quantitative extraction studies.


CONCLUSIONS
5-ALA derivatives differ widely with respect to their enzymatic degradation. The presence of alkyl chains in 5-ALA esters significantly influences the in vitro enzymatic metabolism and the in vivo PpIX formation in healthy hairless mice skins.
De Rosa
Fernanda Scarmato
FS
Department of Pharmaceutical Sciences, Faculty of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, São Paulo, Ribeirão Preto, Brazil.
Lopez
Renata Fonseca Vianna
RF
Thomazine
José Antônio
JA
Tedesco
Antônio Cláudio
AC
Lange
Norbert
N
Bentley
Maria Vitória Lopes Badra
MV
D000622:Aminolevulinic Acid / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism*; D000818:Animals; D004952:Esters; D008297:Male; D051379:Mice; D008812:Mice, Hairless; D017319:Photosensitizing Agents / Q000378:metabolism; D011524:Protoporphyrins / Q000032:analysis / Q000378:metabolism*; D012867:Skin / Q000737:chemistry / Q000378:metabolism*
D004952:Esters; D017319:Photosensitizing Agents; D011524:Protoporphyrins; D000622:Aminolevulinic Acid; C028025:protoporphyrin IX