A water-soluble C60-porphyrin compound for highly efficient DNA photocleavage

A water-soluble covalently linked C(60)-porphyrin compound-C(60)Por is reported to show enhanced DNA-cleaving activity. The highly enhanced DNA-cleaving activity is expected to be ascribed to the high water-solubility and affinity of C(60)Por to DNA, as well as to the accessibility to the charge-separated states C(60)˙(-)-Por˙(+) formed under radiation.     

A potential water-soluble ytterbium-based porphyrin-cyclen dual bio-probe for Golgi apparatus imaging and photodynamic therapy

An amphiphilic water-soluble ytterbium complex which is photo-selective, functional and Golgi apparatus specific has been designed as a responsive dual probe capable of sensitizing emission within the biological window (660 and 750 nm), generating the singlet oxygen (Φ(Δ) = 0.45) and triggering local cell damage only upon exposure to visible and near-infrared laser excitation.     

Induction of cell death by photodynamic therapy with water-soluble lipid-membrane-incorporated [60]fullerene

Intracellular uptake of a lipid-membrane-incorporated C(60) with a cationic surface into HeLa cells was found to induce cell death under visible light irradiation in high efficiency.     

Chlorofullerene C60Cl6: a precursor for straightforward preparation of highly water-soluble polycarboxylic fullerene derivatives active against HIV

We report for the first time the application of chlorofullerene C60Cl6 as a substrate for straightforward preparation of highly water-soluble fullerene derivatives, promising compounds for investigation of the biological action of fullerenes in vitro and in vivo. Methyl esters of phenylacetic and benzylmalonic acids were used as reagents in the Friedel-Crafts arylation of C60Cl6 that resulted in the corresponding C60(Ar)5Cl compounds with 50-60% yields. The following cleavage of ester groups in phenylacetic and benzylmalonic residues was accomplished almost quantitatively to yield the corresponding fullerene-based acids bearing 5 and 10 carboxylic groups, respectively. The relatively-low solubility of these acids in water can be strongly enhanced (up to 150-200 mg ml(-1)) by their conversion to salts with alkali metal cations. These fullerene salt derivatives showed pronounced anti-HIV action and low toxicity; these two findings point to the necessity for further investigation of the biological properties of the here-reported compounds.     

Photo-properties of a silicon naphthalocyanine: a potential photosensitizer for photodynamic therapy

Abstract— The photoproperties of derivatized silicon naphthalocyanine have been investigated in order to assess its potential as a photosensitizer for photo-dynamic therapy. Absorption, fluorescence, phosphorescence and triplet absorption spectra have been measured. Oxygen quenching of the triplet state formed singlet oxygen in significant yields.     

Preparation of a water-soluble fluorinated zinc phthalocyanine and its effect for photodynamic therapy

An amphiphilic fluorinated phthalocyanine, zinc tetracarboxyoctafluorophthalocyanine (ZnC4F8Pc) was synthesized and characterized. Its photodynamic efficiency for HeLa cells was compared with hydrophilic zinc octacarboxyphthalocyanine (ZnC8Pc) and hydrophobic zinc hexadecafluorophthalocyanine (ZnF16Pc). ZnC4F8Pc had a remarkable photodynamic effect among the phthalocyanines used. The effect is apparently caused by the fact that ZnC4F8Pc is mainly accumulated in the hydrophobic lipid membrane and is in the photoactive monomer form in HeLa cells.     

Water-soluble self-assembled butadiyne-bridged bisporphyrin: a potential two-photon-absorbing photosensitizer for photodynamic therapy

We have synthesized a novel, two-photon-absorbing photosensitizer for two-photon-absorption photodynamic therapy (2PA-PDT). The molecule is a butadiyne-bridged porphyrin dimer terminated with two water-soluble porphyrin monomers connected through Zn-imidazolyl self-assembly and covalently linked through olefin metathesis. It has an effective two-photon-absorption (2PA) cross-section value, sigma((2)), of 33 000+/-4600 GM with 5-ns pulses at 890 nm measured by using the open-aperture Z-scan technique. The compound was found to generate singlet oxygen, cytotoxic for tumor cells in photodynamic therapy (PDT), under 2PA conditions by conducting photobleaching experiments with anthracene-9,10-dipropionic acid sodium salt (ADPA).     

Electrophoretic behavior of a highly water-soluble dendro[60]fullerene

The aim of the present study was to develop an analytical method for measuring amounts of a dendro[60]fullerene (DF) which is a highly water-soluble [60]fullerene derivative. We tried to define a straightforward methodology using capillary zone electrophoresis, a method which, to our knowledge, has not yet been used to that purpose. Preliminary assays showed that DF has almost the same mobility than the electroosmotic flow (EOF) but in the opposite direction. Attempts were carried out to reduce the EOF and positive results were obtained by adding hydroxypropylcellulose to the background electrolyte. In order to define optimal operating conditions, a Taguchi experimental plan was used to study simultaneously the effects of the main parameters that are pH, ionic strength, methanol amount and hydroxypropylcellulose concentration. Two parameters are of the utmost importance as to their effect on the migration time and separation efficiecy: pH and ionic strength whose actions are opposite.     

Diblock copolymer micelles deliver hydrophobic protoporphyrin IX for photodynamic therapy

Polymeric micelles are emerging as an effective drug delivery system for hydrophobic photosensitizers in photodynamic therapy (PDT). The objective of this study was to investigate the formulation of hydrophobic protoporphyrin IX (PpIX) with MePEG(5000)-b-PCL(4100) [methoxy poly (ethylene glycol)-b-poly (caprolactone)] diblock copolymers and to compare their PDT response to that of free PpIX. The photophysical and photochemical properties of the polymeric PpIX micelles were studied by measuring absorbance and fluorescence spectra, PpIX-loading efficiency and stability, the micelle particle size and morphology, as well as singlet oxygen luminescence and lifetime. The spherical micelles have a high PpIX-loading efficiency of 82.4% and a narrow size distribution with a mean diameter of 52.2 +/- 6.4 nm. The cellular uptake of PpIX in RIF-1 cells using PpIX micelles was approximately two-fold higher than that for free PpIX. Free PpIX and PpIX formulated in micelles exhibited similar subcellular localization in or around the cellular plasma membrane, as demonstrated using fluorescence microscopy. In vitro PDT results showed that the PpIX micelles have markedly increased photocytotoxicity over that with free PpIX, by nearly an order of magnitude at the highest light dose used. The micelles alone had no evident phototoxicity or dark toxicity. These findings suggest that MePEG(5000)-b-PCL(4100) diblock copolymer micelles have great potential as a drug delivery system for hydrophobic photodynamic sensitizers.     

Methylene blue-containing silica-coated magnetic particles: a potential magnetic carrier for photodynamic therapy

We present the preparation and characterization of methylene blue-containing silica-coated magnetic particles. The entrapment of methylene blue (MB), a photodynamic therapy drug under study in our group, in the silica matrix took place during the growth of a silica layer over a magnetic core composed of magnetite nanoparticles. The resulting material was characterized by transmission electron microscopy (TEM), light scattering, and X-ray diffraction. It is composed of approximately 30 nm silica spheres containing magnetic particles of 11 +/- 2 nm and methylene blue entrapped in the silica matrix. The immobilized drug can generate singlet oxygen, which was detected by its characteristic phosphorescence decay curve in the near-infrared and by a chemical method using 1,3-diphenylisobenzofuran to trap singlet oxygen. The lifetime of singlet oxygen was determined to be 52 micros (in acetonitrile) and 3 micros (in water), with both values being in good agreement with those in the literature. The release of singlet oxygen (etaDelta) was affected by the encapsulation of MB in the silica matrix, which caused a reduction to 6% of the quantum yield of MB free in solution. The magnetization curve confirmed the superparamagnetic behavior with a reduced saturation magnetization in respect to uncoated magnetic nanoparticles, which is consistent with the presence of a diamagnetic component over the magnetite surface. The result is a single particle platform that combines therapy (photosensitizer) and diagnostic (MRI contrast agent) possibilities at the same time, as well as drug targeting.     

Photostability of bacteriochlorophyll a and derivatives: potential sensitizers for photodynamic tumor therapy

The photostabilities of bacteriochlorophyll a and several of its derivatives, which are of interest as potential sensitizers in photodynamic tumor therapy, were investigated. The pigments were irradiated with light >630 nm in organic solvents (acetone, tetrahydrofuran, pyridine, methanol, ethanol, n-propanol, 2-propanol and toluene) and in aqueous detergent solutions (cetyl-trimethyl-ammonium bromide [CTAB], lauryldimethyl-aminoxide [LDAO] or sodium dodecyl-sulfate [SDS] and Triton X-100 [TX100]). Their stabilities in these different solvents were determined in the presence and absence of an external sensitizer (pyromethyl-pheophorbide a), oxygen, sodium ascorbate and inert gas (Ar) or vacuum. The photodegradation products of bacteriochlorophyll a in acetone solution were isolated, purified by HPLC and analyzed by their absorption spectra and mass spectroscopy. Besides the well-known dehydrogenation products, such as [3-acetyl]-chlorophyll a, which were obtained as by-products, the major products had low absorption in the visible-near infrared spectral range. The spectral signature of the major component of these products was characteristic of linear open-chain tetrapyrroles, but they lacked the characteristic protonation-deprotonation behavior and reactivity of bilins with Zn(++).     

Fluorinated ethylenepropylene copolymer as a potential capillary material in CE

In this work, a new generation UV-transparent polymer, fluorinated ethylenepropylene copolymer (FEP) exhibiting a low degree of crystallinity, extruded in dimensions similar to the most commonly used CE capillaries of approximately 80 mum id and about 360 mum od was investigated for its use as a CE capillary. FEP is transparent down to the low-UV region, and as fluorinated polymers in general are good electrical insulators and exhibit reasonable heat conductivity, it has considerable potential as a material for electrodriven analysis in capillary or microchip formats. The FEP capillary has been characterised with regard to some important aspects for its use as a CE capillary, including its profile of EOF versus pH, as well as procedures for manipulating EOF by coating the inner capillary wall with various semipermanent and dynamic layers. The FEP capillaries were tested and compared with fused-silica capillary for the separation of inorganic and small organic ions using conditions involving direct and indirect detection in the low-UV region. Finally, advantages of the use of the FEP capillary for simultaneous detection of a mixture containing nine inorganic cations and anions using indirect photometric detection with a movable light-emitting diode (LED) detector and a novel electrolyte are demonstrated.     

Toward hypericin-derived potential photodynamic therapy agents

To optimize a hypericin derivative as a potential photodynamic therapy agent its light-induced singlet oxygen/superoxide radical formation capability should be enhanced and its long-wavelength absorption band should be bathochromically shifted to better match medicinal lasers. A heavy-atom-substituted derivative was realized by electrophilic iodination of hypericin to yield 2,5-diiodo-hypericin. Using photodestruction of bilirubin IX alpha this derivative was demonstrated to exhibit an enhanced light-induced singlet oxygen/superoxide radical formation capability as compared to hypericin. With respect to a bathochromically shifted derivative styryl residues were attached to the methyl groups of hypericin by de novo ring synthesis. Although the long-wavelength absorption band of this derivative displayed a bathochromic shift of nearly 40 nm it unfortunately immediately underwent an intramolecular [2 + 2] cycloaddition to yield the corresponding cyclobutane derivative in which the added conjugation system became interrupted.     

Synthesis and functionalization of a highly fluorescent and completely water-soluble poly(para-phenyleneethynylene) copolymer for bioconjugation

Bioconjugation of a highly fluorescent water-soluble poly(para-phenyleneethynylene) (PPE) copolymer with ionic and non-ionic side chains is achieved by means of chain-end modification, providing a design principle for biosensor development.     

Self-assembly of stimuli-responsive water-soluble [60]fullerene end-capped ampholytic block copolymer

A well-defined, water-soluble, pH and temperature stimuli-responsive [60]fullerene (C(60)) containing ampholytic block copolymer of poly((methacrylic acid)-block-(2-(dimethylamino)ethyl methacrylate))-block-C(60) (P(MAA-b-DMAEMA)-b-C(60)) was synthesized by the atom transfer radical polymerization (ATRP) technique. The self-assembly behavior of the C(60) containing polyampholyte in aqueous solution was characterized by potentiometric and conductometric titration, dynamic light scattering (DLS), and transmission electron microscopy. This amphiphilic mono-C(60) end-capped block copolymer shows enhanced solubility in aqueous medium at room and elevated temperatures and at low and high pH but phase separates at intermediate pH between 5.4 and 8.8. The self-assembly of the copolymer is different from that of P(MAA-b-DMAEMA). Examination of the association behavior using DLS revealed the coexistence of unimers and aggregates at low pH at all temperatures studied, with the association being driven by the balance of hydrophobic and electrostatic interactions. Unimers and aggregates of different microstructures are also observed at high pH and at temperatures below the lower critical solution temperature (LCST) of PDMAEMA. At high pH and at temperatures above the LCST of PDMAEMA, the formation of micelles and aggregates coexisting in solution is driven by the combination of hydrophobic, electrostatic, and charge-transfer interactions.     

Synthesis of water-soluble C60-porphyrin hybrid compounds

Water-soluble C60-porphyrin hybrid molecules were first synthesized toward their pharmaceutical applications.     

Pharmacokinetics, tissue distribution and photodynamic therapy efficacy of liposomal-delivered hypocrellin A, a potential photosensitizer for tumor therapy

Hypocrellin A, from Hypocrella bambusae, is a novel photosensitizer of high singlet oxygen quantum yield for photodynamic therapy (PDT). Tissue distributions were studied in tumor-bearing mice as a function of time following administration. The tumor model was S-180 sarcoma transplanted into one hind leg of male Kunming mice; hypocrellin A (HA) was delivered to the mice by intravenous injection of 5 mg/kg of body weight as a suspension either as a unilamellar liposome or in dimethyl sulfoxide (DMSO)-solubilized saline. The HA was isolated from several tissues and organs, as well as tumors and peritumoral muscles and skin. Quantitation was performed by a high-performance liquid chromatographic technique with detection that utilizes the native fluorescence of HA. Independent of the delivery system, the dye was retained in tumors at higher concentrations than in normal tissues, except for kidney, liver, lung and spleen. The dye retention in tumors was high and was vehicle dependent. For the liposomal system, the maximal accumulation in tumor and maximal ratios of dye in tumor versus peritumoral muscle and skin occurred 12 h postinjection; for the DMSO saline system, the maximal ratio occurred earlier, 6 h postadministration. Liposomal delivery improved the selective accumulation of the dye in tumor with higher maximal levels in tumor and higher ratios of tumor-to-muscle and tumor-to-skin. Levels of dye were very low or not detectable in the brain. The PDT efficacy of HA in the liposome and DMSO saline systems was determined by evaluating the tumor volume regression percent. The PDT efficacy of HA in liposomes was highest when light treatment was performed at 12 h postinjection, consistent with the highest retention of HA in tumors. Similarly, the maximal PDT efficacy in DMSO saline was attained at 6 h postinjection, the highest HA retention point in tumor. Moreover, the peak PDT efficacy of HA in liposomes was much higher than that of HA in DMSO saline and even hematoporphyrin monomethylether.     

A highly water-soluble 2:1 beta-cyclodextrin-fullerene conjugate

A novel 2:1 (permethylated-beta-cyclodextrin)-fullerene conjugate has been synthesised; highly soluble in cold water with formation of aggregates, it displays a negative solubility coefficient and has a partition coefficient between octanol and water (Log P = 1.58) in the suitable range for biological studies.     

Enhancing the photodynamic therapy efficacy of black phosphorus nanosheets by covalently grafting fullerene C60

Few-layer black phosphorus (BP) nanosheets show potential application in biomedicine such as photodynamic therapy (PDT), and are therefore commonly used in anticancer therapy and nanomedicine due to being relatively less invasive. However, they suffer from low ambient stability and poor therapeutic efficacy. Herein, C₆₀ was covalently grafted onto the edges of BP nanosheets, and the resultant BP-C₆₀ hybrid was applied as a novel endocytosing photosensitizer, resulting in not only significantly enhanced PDT efficacy relative to that of the pristine BP nanosheets, but also drastically improved stability in a physiological environment, as confirmed by both in vitro and in vivo studies. Such improved stability was due to shielding effect of the stable hydrophobic C₆₀ molecules. The enhanced PDT efficacy is interpreted from the photoinduced electron transfer from BP to C₆₀, leading to the promoted generation of ˙OH radicals, acting as a reactive oxygen species (ROS) that is effective in killing tumor cells. Furthermore, the BP-C₆₀ hybrid exhibited low systemic toxicity in the major organs of mice. The BP-C₆₀ hybrid represents the first BP-fullerene hybrid nanomaterial fulfilling promoted ROS generation and consequently enhanced PDT efficacy.

Covalently grafting C₆₀ molecules onto black phosphorus (BP) nanosheets improves their tumor inhibition rate from 36.6% to 88.2%.