Bioconjugate Chem. 2007, 18, 1185−1193
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Porphyrin-Retinamides: Synthesis and Cellular Studies Martha Sibrian-Vazquez, Timothy J. Jensen, and M. Grac¸ a H. Vicente* Department of Chemistry, Louisiana State University, Baton Rouge, Louisiana 70803. Received February 2, 2007; Revised Manuscript Received March 14, 2007
A series of four porphyrin-retinamides containing either all-trans- or 13-cis-retinoid acid residues, directly linked to the para-phenyl position of meso-tetraphenylporphyrin or via a low-molecular-weight PEG spacer, have been synthesized. The biological properties of these conjugates were evaluated in a model cell line, human HEp2, and in neuroblastoma SK-N-DZ cells, which exhibit moderate expression of retinoic acid receptors and retinoic acidinduced differentiation. The directly linked porphyrin-retinamides were taken up by a greater extent (20-50% more) in SK-N-DZ than in HEp2 cells. However, the PEG-containing conjugates accumulated maximally within both cell lines and approximately by the same amount, probably due to their increased amphiphilicity. Among all conjugates, the porphyrin-PEG-13-cis-retinamide accumulated the most in both cell lines (about 5 times more than the non-pegylated conjugates). None of the porphyrin-retinamide conjugates were toxic toward HEp2 cells at concentrations up to 100 µM, and only the hydrophobic non-pegylated conjugates were moderately toxic to SK-N-DZ cells [IC50 (dark) ) 56-92 µM, and IC50 (at 1 J/cm2) ) 6-8 µM]. All conjugates preferentially localized within cellular vesicles that correlated well to the lysosomes and, in addition, the PEG-containing porphyrinretinamides were also found in the ER.
INTRODUCTION The receptor-mediated transport of drugs into targeted cells and sensitive intracellular sites is a valuable approach aimed at increasing the specificity of drugs for diseased tissue (1-4). Photodynamic therapy (PDT) is a binary modality for the treatment of cancer based on the activation of a tumoraccumulated photosensitizer with light of a wavelength, suitable for the excitation of the photosensitizer and for maximum tissue penetration (5, 6). The excited photosensitizer reacts with molecular oxygen and other surrounding molecules, generating radicals and reactive oxygen species (such as 1O2) that cause irreversible photodamage to tumor tissues. Since 1O2, the main cytotoxic agent generated in PDT, has a very short lifetime in biological systems and a very limited range in tissues ( 50 µM) toward HEp2 cells, and only the hydrophobic
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