William G. Love

In vitro interaction of zinc(II)-phthalocyanine-containing liposomes and plasma lipoproteins

We have studied the interaction of small unilamellar liposomes containing zinc(II)-phthalocyanine (Zn-Pc) with human plasma lipoproteins. High-, low- and very low-density lipoproteins (HDL, LDL and VLDL), were purified from plasma and combined in amounts reflecting their natural abundance in plasma. After short periods of incubation at 37 degrees C, the bulk of Zn-Pc was incorporated into HDL and LDL; very little 14C-labelled palmitoyl oleoyl phosphocholine, the most abundant phospholipid in the formulation, was associated with lipoproteins. When liposomes were incubated in pooled plasma, 73%-85% of Zn-Pc and 27%-34% of radiolabelled phospholipid were recovered with HDL and LDL, indicating a possible role for plasma lipid transfer proteins in the incorporation of phospholipid into lipoproteins. Some Zn-Pc was also found in association with VLDL. The buoyant density of Zn-Pc liposomes increased in a dose-dependent fashion when the particles were incubated with plasma, and it is suggested that this was due, at least in part, to opsonization of liposomes by plasma proteins.

Interaction between zinc(II)-phthalocyanine-containing liposomes and human low density lipoprotein

The interaction of human low density lipoprotein (LDL) and small unilamellar liposomes containing the photosensitiser zinc(II)-phthalocyanine (Zn-Pc) was studied in vitro to determine if Zn-Pc could be directly incorporated into the lipoprotein in the absence of other serum components. Incubation of LDL with increasing concentrations of liposomes resulted in a progressive increase in the net negative charge of LDL as determined by agarose gel electrophoresis and both Zn-Pc and liposomal phospholipid were incorporated into the modified LDL particles. Gel chromatography experiments indicated an increase in the molecular mass of modified LDL and immunoaffinity chromatography provided evidence that apoprotein B epitopes on modified LDL were unable to bind to antibody. The study indicated that the liposomal components could be selectively incorporated into LDL by a process that did not appear to involve either aggregation or fusion of particles.

Uptake of zinc(II)-phthalocyanine by HepG2 cells expressing the low-density lipoprotein receptor: studies with the liposomal formulation CGP55847

Hydrophobic photosensitizers readily intercalate into plasma lipoproteins. Some tumors acquire cholesterol from the circulation as a result of increased low density lipoprotein (LDL) receptor activity. Thus, circulating LDL may function as a vehicle for the delivery of bound Zn-Pc to cells within a tumor. Zn-Pc:LDL complexes, resulting from the interaction of LDL with the liposomal Zn-Pc formulation CGP55847, bind to the LDL receptor expressed on HepG2 cells but with reduced affinity in comparison to LDL. Confocal fluorescence microscopy facilitated the subcellular localization of Zn-Pc in microcolonies of HepG2 cells; the photosensitizer was distributed throughout the cellular membrane systems but was absent from the cell nucleus. Uptake of Zn-Pc in the presence of LDL was twofold greater than in the absence of the lipoprotein. These data suggest that the LDL uptake pathway may contribute to the localization of Zn-Pc in hyperproliferative tissue.

Porphyrin self-assembled monolayers and photodynamic oxidation of tryptophan

The zinc and magnesium metalated derivatives of 5,5′-[12,12′-di(thiododecyloxy)-4,4′-phenyl)]-10,10′,15,15′,20,20′-hexakis(3,3′,4,4′,5,5′-hexakisdecyloxyphenyl)diporphyrin, 1b and 1c, have been synthesised and deposited to form self-assembled monolayer (SAM) films on the surface of gold-coated glass substrates. The SAM films have been characterized by RAIR spectroscopy and fluorescence spectroscopy. The potential for the porphyrin films to catalyze the oxidation of tryptophan within human serum albumin upon irradiation with white light has been demonstrated and attributed to the porphyrins acting as photosensitizers of oxygen to form oxidizing species.

Synthesis and biological evaluation of a series of new germanium phthalocyanines incorporated into liposomes--part II: biological evaluation

The pharmacokinetic and phototherapeutic properties of new phthalocyanines with Ge(IV) or Si(VI) as the central metal ion and cholesterol, cholestan or long-chain fatty acids residues as axial ligands to the central ion have been studied in tumor-bearing mice. The new photosensitizers were selectively taken up by and relatively quickly released from the tumors. Except for Si(IV)-Pc, which showed a comparably high selectivity for tumor versus peritumoral tissue, all Ge(IV)-Pc were less selective than liposomal Zn-Pc (CGP 55847). However, all the new compounds showed excellent phototherapeutic efficiency at very low drug and light doses in studies in Meth-A-sarcoma-bearing mice.

Localization of zinc(II)-phthalocyanine within implanted tumors after intravenous administration of a liposomal formulation

The liposomal zinc(II) phthalocyanine (Zn-Pc) formulation CGP55847 was administered intravenously (0.5 mg Zn-Pc/kg) to C57/BL6 mice bearing subcutaneously implanted B16 melanomas or to Swiss mice bearing intramuscularly implanted Ehrlich carcinomas. Tumors were removed 3 or 24 h after dosing, and the Zn-Pc content and intratumoral distribution determined by extraction and quantitative fluorescence microscopy. Localization of the photosensitizer within the tumor mass occurred more rapidly in the highly vascularized Ehrlich carcinoma compared to the less highly vascularized B16 melanoma. Zn-Pc was evident in and around blood vessels 3 but not 24 h after dosing. More Zn-Pc was found in necrotic areas compared to viable tumor tissue; little or no Zn-Pc was detected in the muscle tissue invaded by the Ehrlich carcinoma. At the cellular level, Zn-Pc was associated with membranes and the cytosol but not the nucleus.

Tissue distribution of liposomal zinc(II)-phthalocyanine in normolipaemic and hyperlipaemic rats following intravenous administration

Liposomal formulations of the photosensitizer zinc(II)-phthalocyanine (Zn-Pc) readily interact with plasma lipoproteins in vitro and in vivo, leading to a redistribution of the photosensitizer amongst the major lipoprotein classes. As lipoprotein binding may facilitate the uptake of lipophilic photosensitizers by proliferating tissues, we have examined the fate of intravenously administered liposomal Zn-Pc in normolipaemic rats and rats rendered hyperlipaemic through dietary intervention. Differences were found in the plasma decay profiles of Zn-Pc and liposomal phospholipid between the two groups of animals.