Composite phospholipid-gold nanoparticles with targeted fragment for tumor imaging.

Gold nanoparticles and their conjugates have significant potential in the field of diagnosis of various diseases due to their SPR, which enhances light scattering and absorption. Conjugates of gold nanoparticles with various ligands can be used for imaging biomolecules or detecting malignant neoplasms at an early stage. This study focuses on the construction of composite (or hybrid) phospholipid-gold nanoparticles using soy phosphatidylcholine and a targeted ligand (folic acid derivative) to attach specific targeting properties. According to the method of dynamic light scattering, the diameter of the obtained nanoparticles was less than 100 nm, the results of the MTT test indicated their moderate cytotoxicity. In vitro and in vivo experiments showed a significant increase in the accumulation of phospholipid-gold nanoparticles with a targeted fragment compared to those without a targeted fragment both in HeLa cells and in a tumor (in BDF mice with an injected LLC tumor). The resulting nanoparticles are suitable for specific delivery into tumor cells and visualization of various malignant neoplasms, including at early stages, due to the increased expression of the folate receptor characteristic of cells of a wide range of tumors.

[Improving of HDL capacity for macrophages cholesterol efflux after plasma incubation with phospholipid nanoparticles]. / Povyshenie sposobnosti lipoproteinov vysokoi plotnosti k vyvedeniiu kholesterina iz makrofagov pri inkubatsii plazmy s ul'tramalymi fosfolipidnymi chastitsami.

In connection with recent data about antiatherogenic importance of not only plasma HDL concentration, but of their cell cholesterol efflux capacity as well, the possibility of its correction by phospholipid (PL) nanoparticles was studied. Blood plasma was incubated with earlier elaborated PL nanoparticles emulsion with the particle diameter up to 30 nm, and HDL cholesterol efflux capacity of apo B-depleted plasma was studied. Using macrophages THP-1 preloaded 3H-cholesterol were used. The addition of incubated plasma supernatants with the elevated PL/apo A-1 ratio to cell media resulted in almost increase in two fold 3H-cholesterol efflux as compared with native HDL. The maximal efflux was observed at the PL/apo A-1 ratio of 1.06 as compared with native apo B-depleted plasma (the PL/apo A-1 ratio of 0.85). Results suggest possible usage of ultrasmall PL nanoparticles for regeneration of impaired antiatherogenic HDL functionality. This approach seems to be predominant compared with the usage of PL emulsions with detergent or apoprotein A1.

[Dysfunctional High-Density Lipoproteins: Role in Atherogenesis and Potential Targets for Phospholipid Therapy].

In recent years the number of articles on damages of high-density lipoproteins (HDL) properties in patients with atherosclerosis has sharply increased. First, it concerns their ability to accept cholesterol (CH) from macrophages - the basis of antiatherogenic action of HDL. This ability was assessed ex vivo - by activity of cell cholesterol (CH) efflux to HDL or into patient's serum. In many works inverse relationship was shown between CH acceptor capacity of HDL and severity of atherosclerotic disease or frequency of its exacerbations during long-term observation, independent from HDL CH concentration. This led to the emergence of the concept of importance of "not only HDL quantity but also of their quality", i. e. functionality. In this review we consider pathways of cellular CH efflux (mainly mediated by cell proteins), methods used for detection of dysfunctional HDL, and results of relevant studies in various categories of patients. These studies directed to identification of mechanisms of damages of HDL properties by means of analysis of their composition, used various approaches including those of proteomics and lipidomics. However, now there are no proven targets for correction of HDL dysfunctionality. The only factor, that is underlined by many authors, is the significance of HDL phospholipids, which level correlates with activity of cellular CH efflux. This allows to take a fresh look at previously used phospholipid therapy of atherosclerosis. Its mechanism is apparently not lowering of plasma CH, as was previously expected, but the improvement of HDL antiatherogenic properties. For its practical usage it is necessary to elaborate principally novel formulations with high bioavailability of phospholipids - for HDL enrichment by phospholipid and thereby normalization of their ability to remove CH from tissues.

[Pharmacological targets for dislipidemies correction. Opportunities and prospects of therapeutic usage]. / Farmakologicheskie misheni korrektsii dislipidemii. Vozmozhnosti i perspektivy terapevticheskogo ispol'zovaniia.

Literature data on influence of existing and new groups of drug preparations for dyslipidemias correction are systemized, and molecular mechanisms of their effects are reviewed. The results of experimental and clinical investigations aimed at revealing of new pharmacological targets of dyslipidemias correction were analyzed. The approaches for activation of high density lipoproteins functionality are described. The implementation of alternative preparations with new alternative mechanisms of action may be suggested to improve the effectiveness of traditional treatment in the future.

[Influence of doxorubicin inclusion into phospholipid nanoparticles on tumor accumulation and specific activity]. / Vliianie vkliucheniia doksorubitsina v fosfolipidnye nanochastitsy na nakoplenie v opukholi i spetsificheskuiu aktivnost'.

The specific activity of drug formulation of doxorubicin embedded into phospholipid nanoparticles with diameter less than 30 nm ("Doxolip") was studied in mice LLC carcinoma. Doxolip was prepared according to technology that was elaborated in Institute earlier. Doxorubicin tumor accumulation after intraperitoneal administration (at 4 h) was 4.5 times higher for Doxolip, than for free doxorubicin. The study of doxorubicin antitumor activity in developing tumor after single intravenous administration, 48 h after inoculation, showed, that: 1) tumor growth inhibition of Doxolip was observed at 6th day, while it was only at 11th day for free doxorubicin and revealed in less extent; 2) there was no antitumor effect of free doxorubicin at 8 days after administration of doses 2 and 4 mg/kg, but it was observed for Doxolip in dose-dependent manner, 10% and 30% correspondently. In experiment with developed tumor weekly Doxolip intraperitoneal administration (5 mg/kg, 3 weeks beginning from 7 days after inoculation) resulted in 56% decrease of tumor volume as compared with control. This parameter for free doxorubicin was 2.8 times lower. The obtained data indicate, that incorporation of doxorubicin into phospholipid nanoparticles with size up to 30 nm as delivery system increases its tumor accumulation and results to increase of specific activity both in intraperitoneal and in intravenous administration.

[Investigation of efficacy of phospholipovit for correction of the hepatic encephalopathy]. / Issledovanie éffektivnosti preparata fosfolipovit dlia korrektsii pechenochnoi éntsefalopatii.

This paper presents the results of clinical studies on the efficacy and safety of the drug Phospholipovit in different groups of patients, in particular with hepatic encephalopathy and with a high risk of its development (chronic alcohol intoxication). Efficacy of treatmernt was evaluated by the LNT test (link numbers test), and standard liver plasma markers (ALT, AST, GGT, AP). The LNT test in patients with encephalopathy showed better improvement after 5 days course of Phos-pholipovit than after standard therapy. In both clinical trials liver enzyme activities, assayed in pa-tients declined more rapidly in the group of patients treated with Phospholipovit, as compared in patients received standard therapy alone. The highest clinical effect of the drug on the liver function tests was observed at a daily dose of 6.4 g of phospholipids (infusional 2 times a day) for 5-10 days. At the end of treatment a two-fold decrease in the activity of AST was observed in patients receiv-ing Phospholipovit compared to the control. This results of clinical results can be regarded as a manifestation of the expressed membrane repairing action of essential phospholipids, reinforced by their introduction into the body in the form of nanoparticles.

[Pharmacological agents and transport nanosystems based on plant phospholipids].

A new generation of plant phosphatidylcholine (PC)-based pharmacological agents has been developed under academician A.I. Archakov leadership at the Institute of Biomedical Chemistry (IBMC). For their production a unique technology allowing to obtain dry lyophilized phospholipid nanoparticles of 30 nm was elaborated. The successful practical application of PC nanoparticles as a drug agent may be illustrated by Phosphogliv (oral and injection formulations). Being developed at IBMC for the treatment of liver diseases, including viral hepatitis, Phosphogliv (currently marketed by the "Pharmstandard" company) is approved for clinical application in 2000, and is widely used in medical practice. Based on the developed and scaled in IBMC technology of prerparation of ultra small size phospholipid nanoparticles without the use of detergents/surfactants and stabilizers another drug preparation, Phospholipovit, exhibiting pronounced hypolipidemic properties has been obtained. Recently completed preclinical studies have shown that PC nanoparticles of 20-30 nm activate reverse cholesterol transport (RCT) and in this context it is more active than well known foreign preparation Essentiale. Phospholipovit is now at the stage of clinical trials (phase 1 completed). PC was also used as a basis for the development of a transport nanosystem with a particles size of 20-25 nm in diameter and incorporation of various drug substances from various therapeutic groups. Using several drugs substances as an example, increased bioavailability and specific activity were demonstrated for the formulations equipped with such transport nanosystem. Formulations equipped with the transport nanosystems have been developed for such pharmacological agents as doxorubicin, rifampin, budesonide, chlorin E6, prednisone, and others.

[The pathogenesis of surgical trauma and the characteristics of its course in planned surgical interventions in the maxillofacial area]. / Patogenez operatsionnoi travmy i osobennosti ee techeniia pri planovykh khirurgicheskikh vmeshatel'stvakh v cheliustno-litsevoi oblasti.

Examinations carried out on 76 patients have shown that systematic surgical interventions into the maxillofacial area produce deep changes in the tissue and serum systems of proteolysis and their inhibitors. Activation of proteolytic enzymes already on the stage of surgical trauma leads to the formation in the operative wound of the focus of endotoxicosis which depresses antitryptic activity of injured tissues and thus promotes the development of purulent complications. The study of the above indexes at the early stages of acute period may serve as a diagnostic test while estimating a severity of operative wound and allows to make prognosis of the development of purulent complications.