Biomedical subjects

P Caliceti

Publications and source records attributed to P Caliceti.

At least 37 records · Page 2Linked to original sources

PubMed1996

Covalent modification of mushroom tyrosinase with different amphiphic polymers for pharmaceutical and biocatalysis applications.

Two different poly(ethylene glycol) derivatives (linear, mol wt 5000 and a branched form, mol wt 10000) and a new polymer (poly-[acryloylmorfoline], mol wt 5500) were covalently bound to the enzyme tyrosinase. The polymer-protein conjugates were studied with a view to their potential pharmaceutical application and to their use for the bioconversion of phenolic substrates in organic solvents. Vmax and Km for the dopa-dopaquinone conversion, thermostability, stability toward inactivation by dopa oxidation products, half-life in blood circulation, and behavior in organic solvents for the different adducts were investigated. Arrhenius plots for the dopa-dopaquinone conversion were also obtained in order to study the effects of temperature on the different enzyme forms. Covalent attachment of the polymers increased enzyme stability in aqueous solution and the solubility in organic solvents. However, organic solvent solubilization brought about loss of enzyme conformation as assessed by CD measurements, which is accompanied by a nonreversible loss of catalytic activity.

Animals↗

PubMed1996

Anti-inflammatory activity of polyphosphazene-based naproxen slow-release systems.

A biocompatible and biodegradable polyphosphazene bearing phenylalanine ethyl ester, imidazole and chlorine (10.7:1:2.5 molar ratio) as substituents of the phosphorus atoms of the polymer backbone was studied for the preparation of polymeric naproxen slow-release systems. Discs 2.5 cm in diameter and 0.5 mm (thin) or 0.65 mm (thick), loaded, respectively, with 20 and 13.5% naproxen, showed different drug release kinetics, the thin matrices releasing naproxen at a faster rate and for a shorter time. In-vivo studies in rats demonstrated the pharmacological efficacy of these two different delivery systems in the inhibition of acute or chronic inflammatory diseases. Subcutaneous implantation of the thin matrices in rats was found to reduce carrageenan oedema induced both 1 h and 7 days after implantation. Rats implanted with thick matrices showed a reduction in chronic inflammation caused by adjuvant arthritis. Approximately 78% inhibition of arthritic oedema was found 28 days after subcutaneous administration of the matrices whereas 28.7% inhibition was found after daily oral administration of naproxen. Blood levels of naproxen in arthritic rats after matrix implantation showed the presence of drug up to day 28. These positive results have encouraged us to study a controlled-release system suitable for use in man.

Administration, Oral↗

PubMed1996

Pathophysiological and histomorphological evaluation of polyacryloylmorpholine vs polyethylene glycol modified superoxide dismutase in a rat model of ischaemia/reperfusion injury.

Twenty Wistar rats were divided into two groups. Both underwent acute ischaemia followed by reperfusion of the left hind limb. The first group was a control group while the second was treated with PAcM-SOD. The survival percentage of the limb after 10 days was 30% for the first group and 70% for the second. Neither linear regression nor correlation were found between groups as far as the survival percentage of the limb after 10 days and reperfusion pmO2 data were concerned. After ten days the histomorphological analysis was significant regarding the fibre diameter and the percentage of central located nuclei in the specimens of PAcM-SOD treated limbs compared to normal limbs, but not when compared to the muscular fibres of the control group. Comparing these results with others obtained with native SOD and monomethoxypoly(ethylene glycol) modified SOD (mPEG-SOD) used in the same experimental model, we can conclude that the clinical and morphological results were better using mPEG-SOD, and that PAcM-SOD does have a protective effect on ischaemic muscle damage, although it is not as effective as mPEG-SOD in preventing ischaemia/reperfusion injury.

Acrylic Resins↗

PubMed1995

New synthetic amphiphilic polymers for steric protection of liposomes in vivo.

Carboxy group-terminated synthetic polymers--branched poly(ethylene glycol), poly(acryloylmorpholine), and poly(vinylpyrrolidone)--were made amphiphilic by derivatization with phosphatidyl ethanolamine via the terminal carboxy group and then incorporated into lecithin-cholesterol liposomes prepared by the detergent dialysis method. Following the biodistribution of liposomes in mice, all three polymers were shown to be effective steric protectors for liposomes and were able to sharply increase liposome circulation times in a concentration-dependent manner. The accumulation of liposomes in the liver decreases. The effects observed are similar to those found for liposomes modified with linear poly(ethylene glycol). At low polymer concentration, amphiphilic branched poly(ethylene glycol) seems to be the most effective liposome protector, most probably, because at the same molar content of anchoring groups, each attachment point carries two polymeric chains and doubles the quantity of liposome-grafted polymer comparing to linear poly(ethylene glycol).

Animals↗

PubMed1995

Peripheral nerve repair using a poly(organo)phosphazene tubular prosthesis.

Nerve regeneration experiments were carried out using tubular nerve guides of poly[(ethylalanato)1.4(imidazolyl)0.6phosphazene] (PEIP). By means of in vivo tests, this polymer was found to be biodegradable and transformed into harmless products. The tubular nerve guides were prepared by deposition of the dissolved polymer on a glass capillary tube, followed by evaporation of the solvent (methylene dichloride). After transectioning, rat sciatic nerve stumps were immediately sutured into the ends of 10-mm-long polymer tubes. On removal of the prosthesis, after implantation for 45 d, a tissue cable was found bridging the nerve stumps in all cases. Histological analysis revealed that the tissue cable was essentially composed of a regenerated nerve fibre bundle. A parallel series of experiments was undertaken to compare the use of silicone tubes that are not biodegradable and are most frequently used for studies of nerve regeneration with tubulization techniques. The advantages of biodegradable PEIP tubular nerve guides used for peripheral nerve repair are discussed.

Animals↗

PubMed1995

Biopolymeric modification of superoxide dismutase (mPEG-SOD) to prevent muscular ischemia-reperfusion damage.

The efficacy of preventing ischemia-reperfusion damage by employing native or modified (mPEG-SOD) superoxide dismutase in an experimental animal model of acute ischemia of the left hindlimb was tested. Four hours and thirty minutes complete warm ischemia was induced in the left hindlimb of 43 Wistar rats, by clamping the femoral artery and monitoring its efficacy with Laser Doppler flowmetry. After ten days, a significative difference (p = 0.004) of the survival leg rate was found in the group treated with mPEG-SOD (86.6%) compared with the control group (30%). Histomorphological and ultrastructural analysis were performed at different time intervals confirming what the clinical course had already pointed out. These results show that SOD in its modified form, despite the lower dosage, can provide good protection against ischemia/reperfusion injury of muscles.

Animals↗

PubMed1995

Preparation and characterisation of polyphosphazene-based controlled release systems for naproxen.

A new polyphosphazene polymer suitable for preparation of naproxen controlled release systems has been prepared by non complete substitution of the chlorine at the phosphorus atoms of polydichlorophosphazene with phenylalanine ethyl ester and imidazole. Polymeric disks obtained by solvent evaporation were found to maintain their integrity for months either "in vivo" and "in vitro" after incubation in buffer, although the intrinsic viscosity undergoes a rapid decrease. The release rate of naproxen from the films was found to depend on thickness of the matrixes and amount of entrapped drug. "In vitro" release spikes, corresponding to initial in plasma burst after subcutaneous implantation of the films in rats, have been observed with high concentrations of drug in the matrixes. On the other end decreasing the drug concentrations in the polymer matrixes, the naproxen release approximates a zero order kinetic and a controlled release is obtained for weeks. Scanning electron microscopy showed that the disks giving spikes in drug release are characterised by the presence of naproxen crystals on the matrix surface. "In vivo" studies are demonstrating that the initial naproxen concentration spike is useful in the treatment of acute models of inflammation, while a more constant and lasting release is successful in chronic inflammation models. The degradation profile of this new polyphosphazene as well as preliminary pharmacological data are also reported.

Animals↗

PubMed1995

Distribution of catalase, ribonuclease and superoxide dismutase modified by monomethoxy (polyethylene glycol) into rat central lymph and lymphatic nodes.

The plasma-lymphatic distribution of ribonuclease (RNase), superoxide dismutase (SODase), and catalase (CTase) modified by monomethoxy (polyethylene glycol) (mPEG) was studied in rats. The lymphatic bioavailability (FL) of individual enzymes administered intravenously was determined on the basis of plasmatic and lymphatic concentration curves. It was concluded that FL values depend on enzyme-adduct molecular weight (m.w.). The highest FL value was found in mPEG-RNase (the lowest m.w.), medium value in mPEG-SODase (intermediate m.w.), and the lowest one in mPEG-CTase (the highest m.w.). The binding of these enzymes in the lymphatic tissue of iliac, intestinal, brachial and neck nodes was also proportional to their molecular weight. The lymphatic binding was dependent on the node localization, higher concentrations being found in the iliac and neck nodes in contrast to the other nodes (intestinal, brachial).

Animals↗

PubMed1994

Radiation grafting of hydrophilic monomers onto poly[bis(trifluoroethoxy)phosphazene].

The hydrophilic monomers dimethylacrylamide (DMAA) and acrylamide (AAm) were radiation grafted onto poly[bis-(trifluoroethoxy)phosphazene] (PTFP) for biocompatibility enhancement through direct irradiation of the polymer films immersed in monomer-solvent mixtures, and the progress of grafting was followed up to a large monomer depletion. The water absorption of the samples was found to increase as the amount of the grafted chains increased. The grafted chains, mainly confined in the film surface at high dose rates, progressively extend to the interior of the sample as the dose rate decreases. In the grafted samples the ultimate mechanical properties epsilon r and sigma r were essentially preserved. The in vivo tests indicated an enhancement of surface biocompatibility in the PTFP-g-AAm samples and not in the PTFP-g-DMAA samples.

Absorption↗

PubMed1993

Pharmacokinetics and distribution of ribonuclease and its monomethoxypoly(ethylene glycol) derivatives in rats.

The changes of pharmacokinetics and distribution of 3H radio-labelled ribonuclease (RNase) and RNase adduct with monomethoxypoly(ethylene glycol) (MPEG) were studied after intravenous administration in the dose of 25 mg kg-1 to rats. Whereas the value of total plasma clearance of RNase is close to the value of glomerular filtration rate, the value of that for MPEG-RNase is about three hundred times lower. The half-life of elimination is 79 min, 181 min and 65 h for RNase, MPEG and MPEG-RNase, respectively. The main elimination pathway of compounds under study was elimination into urine and no evident specific distribution in the examined organs (liver, kidney, muscle, lung, brain, spleen and heart) was found. The study indicates that conjugation of RNase with MPEG can improve its pharmacokinetic properties.

Animals↗

PubMed1993

Polyphosphazenes as biomaterials: surface modification of poly(bis(trifluoroethoxy)phosphazene) with polyethylene glycols.

Investigations were carried out on the metathetical exchange reaction between the -O-CH2CF3 moieties of poly(bis(trifluoroethoxy)phosphazene) (PTFP), in the state of slightly swollen films, and the alkoxide ions derived from methoxypolyethylene glycol (MPEG) of molecular mass ranging from 350 to 5000 g/mol. The substitution of these hydrophilic chains, mostly confined to thin surface layers, was revealed by means of optical microscopy and scanning electron microscopy observations, surface elemental analysis by energy-dispersive X-ray analysis (EDXA), FTIR-ATR analysis and water contact angle measurements. The surface biocompatibility was enhanced in all cases, whilst the mechanical properties of the original PTFP films were substantially retained in the modified samples exhibiting low substitutions. Such samples were obtained especially when the metathetical reaction was carried out with MPEG5000.

Animals↗

PubMed1993

Preparation and properties of monomethoxy poly(ethylene glycol) doxorubicin conjugates linked by an amino acid or a peptide as spacer.

Polymeric doxorubicin prodrugs were prepared linking monomethoxy poly(ethylene glycol), 5000 D molecular weight, to the doxorubicin amino group, using an amino acid or a peptide as a spacer arm. As spacers glycine, L-phenylalanine, L-tryptophan and glycil-L-valil-L-phenylalanine were used. The conjugates showed enhanced stability to alkaline degradation compared to the free doxorubicin. Towards Ehrlich solid tumor in mice the glycin spaced derivative was devoid of activity, whereas the phenylalanine and tryptophan derivatives were 20% and 16% active and the tripeptide one 50% active with respect to free doxorubicin. On the other hand the derivatization was accompanied by a great decrease of toxicity in mice with respect to the free drug. Doxorubicin was not released from conjugates by chymotrypsin incubation or in plasma.

Amino Acid Sequence↗

PubMed1992

Desferrioxamine potentiated SOD antiinflammatory activity in rat adjuvant arthritis: role of iron and bacterial toxins.

In this paper we studied the modulating inflammatory activity of iron in the adjuvant arthritis, taking indomethacin as a standard antiinflammatory drug and a superoxide dismutase derivative (MPEG-SOD) as a scavenger of free radicals. Moreover, we evaluated the changes in potential intestinal pathogens requiring iron for growth, in order to study the role of bacteria in the altered gastrointestinal functions observed during arthritis. We observed a 50% arthritis inhibition on the 14th day with MPEG-SOD plus desferrioxamine, a significant decrease in serum iron in arthritic rats compared to controls, and a significant Cl. perfringens increase on the 28th day in the presence of MPEG-SOD. Our data demonstrate that hypoferremia, in arthritis, is a protective mechanism overall in the early phase and could protect the intestinal tract by inhibiting the development of potential pathogens.

Animals↗

PubMed1992

Controlled release of proteins and peptides from hydrogels synthesized by gamma ray-induced polymerization.

Hydrogels prepared by radiation-induced polymerization at a low temperature have been used as carriers for the controlled release of peptides and proteins. It was found that polymerization of 2-hydroxyethyl methacrylate in the presence of poly (ethylene glycol) methyl ether (MPEG) enabled the more porous and swellable matrics to be obtained, the higher the molecular weight of MPEG. As a consequence, protein release took place at an increasing extent and, provided that MPEG molecular weight was high enough, high molecular weight proteins could also be released. Such a state of affairs was not met in the case of hydrogels based on poly (2-hydroxyethyl acrylate). SEM analysis revealed that even high molecular weight MPEG did not give rise to any porosity, even though the degree of swelling was very high. As a result, no protein release was observed. It was therefore concluded that control of hydrogel porosity for the controlled release of large proteins is of overwhelming importance.

Alcohol Dehydrogenase↗

PubMed1991

Determination of narciclasine in serum by reversed-phase high-performance liquid chromatography: comparison of amperometric, ultraviolet photometric and fluorescence detection.

Narciclasine was determined in the blood of mice by reversed-phase high-performance liquid chromatography, using a C18 stationary phase and a mobile phase of methanol-0.025 M potassium dihydrogen phosphate (50:50, v/v) of pH 5.5. Amperometric detection at a carbon fibre array working electrode held at +1.8 V (Ag/AgCl) permitted determination down to concentrations of 10 and 15.4 ng ml-1 (at a signal-to-noise ratio of 2) in aqueous solution and in serum, respectively. Fluorescence detection (excitation and emission wavelengths of 360 and 480 nm, respectively) exhibited somewhat poorer sensitivities for aqueous and serum samples: the respective limits of detection were 25 and 32 ng ml-1 at a signal-to-noise ratio of 2. Both the amperometric and the fluorescence detection were free from interference from blood components, but the fluorescence measurement required a post-column pH adjustment. UV photometric detection at 254 nm exhibited detection limits of 15 and 65 ng ml-1 in aqueous samples and in serum, respectively, and suffered from interferences from blood components that strongly absorbed in the ultraviolet region. All three detection techniques exhibited good linearity and precision.

Alkaloids↗

PubMed1991

Enzyme modification by MPEG with an amino acid or peptide as spacer arms.

A method for the modification of enzymes by MPEG carrying an amino acid or peptide as a spacer arm is described and tested with aliphatic or aromatic side chains amino acids. The procedure involves MPEG activation by p-nitrophenylchloroformate for the amino acid or peptide coupling that is in turn activated for the protein binding. The advantage of the method resides in the possibility to introduce proper reporter groups between the polymer and the protein as norleucine for a direct evaluation of the bound polymer chains, tryptophan for structural studies of the polymer-protein adduct, and radioactive amino acid for pharmacokinetic investigations. The method was positively tested with arginase, ribonuclease, and superoxide dismutase as enzymes of therapeutic value.

Amino Acids↗

PubMed1991

Accurate evaluation method of the polymer content in monomethoxy(polyethylene glycol) modified proteins based on amino acid analysis.

To overcome the uncertainty of the colorimetric or fluorimetric method so far employed for the evaluation of monomethoxy(polyethylene glycol) (MPEG) covalently bound to protein, a direct method based on amino acid analysis is proposed. The method exploits the use of MPEG, which was bounded with the unnatural amino acid norleucine (MPEG-Nle). MPEG-Nle was activated at its carboxylic group to succinimidyl ester for the binding to the amino groups of protein. After acid hydrolysis, the amino acid content is evaluated by conventional amino acid analyzer or by reverse-phase HPLC as phenylthiocarbamyl derivative. The number of bound MPEG chains is calculated from the amino acid composition, since one norleucine residue is released from each bound polymer chain. The method was verified with several proteins in comparison with colorimetric ones, also in the case of proteins that contain chromophores in the visible range, such cytocrome C. It was observed that in most of the cases, the colorimetric methods give an overestimation of the degree of protein modification.

Amino Acids↗

PubMed1991

Biocompatible polyphosphazenes by radiation-induced graft copolymerization and heparinization.

Investigations were carried out on the radiation-induced graft copolymerization by direct irradiation of dimethylaminoethyl methacrylate on to poly(bis(trifluoroethoxy)phosphazene) and on to poly (bis(phenoxy)phosphazene). Kinetics of grafting were followed with the polyphosphazenes immersed in monomer - methanol mixtures of various composition. The grafted film samples were quaternized with methyl iodide and, to the produced ammonium group, heparin was ionically bonded with high yield. On the grafted and heparinized-grafted film samples an evaluation of hydrophilicity, mechanical properties, biocompatibility and anticoagulating properties was carried out.

Animals↗