[Prednisolone in phospholipid nanoparticles: prolonged circulation and increased antiinflammatory effect]. / Prednizolon v fosfolipidnykh nanochastitsakh: prolongirovannaia tsirkuliatsiia i povyshennoe protivovospalitel'noe deistvie.

Along with modern new drugs, many therapeutic schemes also include known effective drugs, particularly, glucocorticoids. One of the most distributed of them is prednisolone that has pronounced anti-inflammatory properties. Its disadvantage is short-term circulation, resulting in a number of side effects. For this reason the development of its more effective and safe formulations is carried out. We have obtained the formulation of prednisolone included in nanoparticles from soy phosphatidylcholine with an average diameter of 20 nm. With oral administration to rats and analysis by HPLC an increase in prednisolone maximal concentration in of plasma and the duration of circulation as compared with free drug administration were shown. The experiment with mice with conconavalin A induced inflammation was also carried out: conconavalin A was injected subplantary in an hour after oral administration of both prednisolone formulations in several doses. The index of the inflammatory reaction (determined by the edema degree) was suppressed more effectively in the case of prednisolone in nanoparticles. Maximal suppression (62.2% as compared with 49.6% for free prednisolone) was observed even at a minimal dose (2.5 mg/kg), at which the free drug did not act at all. The results indicate an increase in the efficiency of prednisolone included in phospholipid nanoparticles, that makes it possible to diminish its administered doses and thereby reduce the risk of side effects.

Effect of Cell-Penetrating Arginine Peptide on Interaction of Photosensitizer Chlorin e6 Incorporated into Phospholipid Nanoparticles with Tumor Cells.

We studied the possibility of increasing the efficiency of photodynamic therapy by improving delivery of photosensitizers chlorin e6 into tumor cells. Previous studies showed that incorporation of chlorin e6 onto phospholipid nanoparticles with a diameter <20 nm reduces its cytotoxicity due to accelerated elimination from organs [8]. A heptapeptide R7 synthesized and added to this combination promoted internalization of chlorin e6 into HepG2 cells in comparison with initial nanoparticles without peptide R7. The observed effect of peptide R7 can be explained by activation of endocytosis and/or macropinocytosis (bearing in mind the interaction of arginine with carboxyl groups of e6. The development of this transporting system is a promising trend in photodynamic therapy of cancer diseases.

[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.

[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.

[Increase of antituberculosis efficiency of rifampicin embedded into phospholipid nanoparticles with sodium oleate]. / Povyshenie protivotuberkuleznogo deistviia rifampitsina pri vstraivanii v fosfolipidnye nanochastitsy c oleatom natriia.

The formulation of the antituberculosis drug rifampicin embedded into 20-30 nm nanoparticles from soy phosphatidylcholine and sodium oleate, is characterized by greater bioavailability as compared with free drug substance. In this study higher antituberculosis activity of this formulation was shown. Rifampicin in nanoparticles demonstrated more effective inhibition of M. tuberculosis H37Rv growth: minimal inhibiting concentration (MIC) was twice smaller than for free rifampicin. Administration of this preparation to mice with tuberculosis induced by M. tuberculosis Erdman revealed that after 6 weeks of oral administration the CUF value in lung was 22 times smaller for rifampicin in nanoparticles than for free drug (1.7 un. vs. 37.4 un.). The LD50 value in mice was two fold higher for rifampicin in nanoformulation.

[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.

[The increase in receptor-mediated endocytosis of drugs in the composition of nanoparticles with the address fragment]. / Uvelichenie éndotsitoza protivoopukholevykh lekarstv v sostave nanochastits s adresnym fragmentom.

It is known that disorders in the cell functioning of the organs/tissues is accompanied by increased expression of certain receptors. A modern approach to improve the specificity of the drug accumulation in the affected area is to construct the delivery nanosystems with the address fragments. Active tagged transport may help to reduce the dose of the drug, minimizing the impact on healthy cells and organs (reduced adverse events). This approach is particularly important in oncology because of the high toxicity of the drugs used. In this work we have obtained and characterized the pharmaceutical composition of doxorubicin and chlorine e6 into colloidal nanoparticles with synthesized previously targeted conjugates based on folic acid and biotin. On the cell culture Hep G2 it was shown an increase in the internalization of drugs when they were introduced in the incubation medium in the form of drug compositions with transport nanosystems and targeted fragments.

[The in vivo study of the medicinal composition property of doxorubicin as a part of colloidal nanoparticles with the address fragment].

The use of targeted transport systems for drug delivery is a promising approach to improve pharmacokinetics of drug substances, accumulation in the lesion. In this study we have obtained and characterized the pharmaceutical composition of doxorubicin in colloidal nanoparticles equipped with targeted conjugates based on folic acid and biotin with dodecylamine. The inclusion of the address fragments into colloidal nanopartical was carried out without surface and drug substance modification The accumulation of anthracycline antibiotic doxorubicin in tumor tissue was compared in Lewis lung carcinoma mouse models after intravenous administration of the composition of colloidal nanoparticles with targeted conjugates biotin-dodecylamine and folic acid-dodecylamine or free doxorubicin. It was shown that the doxorubicin accumulation in tumor tissue when administered in drug compositions with targeted fragments are 2 times higher for the folic acid-dodecylamine conjugate and 1.4 times higher for the biotin-dodecylamine conjugate.

[Influence of resveratrol and dihydroquercetin inclusion into phospholipid nanopatricles on their bioavailability and specific activity].

The effects of natural polyphenols, resveratrol (RES) and dihydroquercetin (DHQ), included in phospholipid nanoparticles, have been compared with free substances of RES and DHQ in in vitro and in vivo experiments. Preincubation of healthy donor plasma low density lipoproteins (LDL) with RES or DHQ included in phospholipid nanoparticles caused a more pronounced decrease in Cu2+ induced lipid oxidation compared with the free substances, and reduced the formation of lipid peroxides products. Bioavailabilities of RES and DHQ in phospholipid formulations after oral administration in rats were increased by 1.5-2 times. In an acute hypoxia model in mice prophylactic two-week administration of RES or DHQ phospholipid formulations resulted in 25% increase in survival and 1.5-fold increase in catalase activity in brain homogenates compared to free substances. Using the model of endothelial dysfunction in rats induced by L-NAME it was shown, that RES markedly attenuated the inhibition effect of L-NAME on NO synthesis. RES in phospholipid nanoparticles had the same action at a dose 10 times lower compared to free RES. Load test with resistance (clamping of the ascending aorta for 30 sec) showed that phospholipid formulation of RES possessed more pronounced protective effect due to the stimulation of endothelial NO-synthase.

[The influence of doxorubicin incorporated in phospholipid drug delivery nanosystem on HEPG2 cells proteome].

A phospholipid drug delivery nanosystem with particle size up to 30 nm elaborated at the Institute of Biomedical Chemistry has been used earlier for incorporation of doxorubicin (Doxolip). This system demonstrated higher antitumor effect in vivo as compared with free doxorubicin. In this study the effect of this nanosystem containing doxorubicin on HepG2 cell proteome has been investigated. Cells were incubated in a medium containing phospholipid nanoparticles (0.5 mg/ml doxorubicin, 10 mg/mL phosphatidylcholine). After incubation for 48 h their survival represented 10% as compared with untreated cells. Cell proteins were analyzed by quantitative two-dimensional gel electrophoresis followed by identification of differentially expressed proteins with MALDI-TOF mass spectrometry. The phospholipid transport nanosystem itself insignificantly influenced the cell proteome thus confirming previous data on its safety. Doxorubicin, as both free substance and Doxolip (i.e. included into phospholipid nanoparticles) induced changes in expression of 28 proteins. Among these proteins only four of them demonstrated different in response to the effect of the free drug substance and Doxolip. Doxolip exhibited a more pronounced effect on expression of certain proteins; the latter indirectly implies increased penetration of the drug substance (included into nanoparticles) into the tumor cells. Increased antitumor activity of doxorubicin included into phospholipid nanoparticles may be associated with more active increase of specific protein expression.

[Analysis of proteomic profile changes of zebrafish embryos during exposure to doxorubicin, built-in the phospholipid transport nanosystem].

The proteome profile of Danio rerio embryos grown in the medium containing doxorubicin, included in the phospholipid transport nanosystem (doxolip) has been investigated using combination of 1D-electrophoresis with subsequent MALDI-TOF-PMF mass spectrometry. Cultivation of growing of D. rerio embryos in the medium with doxolip caused a substantial increase in expression of the cytoskeletal proteins, a decrease in the number of nuclear proteins involved in DNA and RNA synthesis and disappearance of vitellogenin 2 in comparison with control (the cultivation medium containing the phospholipid transport nanosystem). Analysis of the proteomic profiles of doxolip-treated embryos suggests lower toxicity of doxorubicin incorporated in the phospholipid nanosystem.

Application of small-angle X-ray scattering to the characterization and quantification of the drug transport nanosystem based on the soybean phosphatidylcholine.

Phospholipid transport nanosystem (PTNS) for drug delivery is based on soybean phosphatidylcholine. The morphology of PTNS is investigated by means of small-angle X-ray scattering. The obtained results allow one to answer the key question from the viewpoint of organization of drug incorporation whether the PTNS nanoparticles have a structure of micelles or vesicles. It is demonstrated that PTNS is a vesicular system with an average vesicle radius of 160 ± 2Å.

[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.

[Mass spectrometry analysis of blood plasma lipidome as method of disease diagnostics, evuation of effectiveness and optimization of drug therapy].

A new method for the analysis of blood lipid based on direct mass spectrometry of lipophilic low molecular weight fraction of blood plasma has been considered. Such technique allows quantification of hundreds of various types of lipids and this changes existing concepts on diagnostics of lipid disorders and related diseases. The versatility and quickness of the method significantly simplify its wide use. This method is applicable for diagnostics of atherosclerosis, diabetes, cancer and other diseases. Detalization of plasma lipid composition at the molecular level by means of mass spectrometry allows to assess the effectiveness of therapy and to optimize the drug treatment of cardiovascular diseases by phospholipid preparations.

[Interaction of rifampicin embedded in phospholipid nanoparticles with blood plasma lipoproteins].

The drug formulations of antituberculous remedy rifampicin in nanoparticles less than 30 nm based on soy phosphatidylcholine and sodium oleate was elaborated in Institute of Biomedical Chemistry. The distribution of rifampicin in blood plasma fractions after incubation with this formulation and with free rifampicin was studied. This goal was stimulated by the literature data about activation of macrophages LDL receptors in cases of M. tuberculosis infection. Plasma was incubated 30 min with free rifampicin or rifampicin encapsulated into the nanoformulation followed by ultracentrifugation and subsequent rifampicin determination by HPLC in lipoprotein fractions. In the case of free rifampicin it appeared mainly in the plasma protein fraction and in HDL (41% and 38%, correspondentely). But after incubation of rifampicin in nanoparticles the drug redistribution was observed. Its proportion in these factions decreased 2-3-fold, and it was found mainly in LDL (60% as compared with 21% for free rifampicin). The increased association of rifampicin encapsulated into phospholipid nanoparticles with LDL is considered as facilitating factor for macrophages delivery and thus for antituberculosis efficiency as well.

Effects of fullerene C60 on proteomic profile of Danio rerio fish embryos.

The effects of phosphatidylcholine-based phospholipid nanoparticles containing fullerene C60 on Danio rerio fish embryos were studied. Exposure of the embryos with the nanoparticles for 48 h did not lead to appreciable changes in the number of protein bands in SDS-PAGE in comparison with the control (exposure in medium with phosphatidylcholine). Mass spectrometric identification of proteins showed differences in the proteomic profiles of the samples. The content of vitellogenins changed after exposure with phosphatidylcholine-based nanoparticles with C60 fullerenes. This could indicate low toxicity of the nanoparticles towards D. rerio embryos under experimental conditions.

[Development of targeted drug delivery system: synthesis of conjugates of address fragment (RA-COOH) with ligand (R-NH2)].

One of the main ways to increase the effectiveness of well-known medical formulations well-established in clinical medicine - development of delivery systems using new technological approaches and nanomaterials. Currently, much attention is given to targeted delivery systems. At the same time drug carrier has in addition to medication the so-called vector/address with a high affinity for binding to specific receptors on cells/tissue target. In this paper it is described the method for producing of address conjugates to over-expressed receptors on the tumor cells. As address fragment it was folic acid and as a linker was dodecylamine, causing inclusion the conjugate into lipid nanoparticles.

[Nanoparticles as drug delivery system for antituberculous drugs].

The increase of tuberculosis incidence in last decade stimulated elaboration of both new antituberculous drugs and also searches ofoptimiting delivery systems for existing drugs. It is determined by their side effects and low bioavailability of effective first line drug rifampicin. Various nanosystems for transport of antituberculous drugs are considered on the basis of various polymers, liposomes, lipid nanoparticles, nanoemulsios, nanosuspensions, dendrimers, cyclodextrines. Influence of drug incorporation into nanoparticles, most often for rifampicin, on pharmacokinetics and efficiency in tuberculosis models is discussed. The most of works are devoted to polymer nanoparticles for oral administration where increased circulation time and efficiency were shown. The best results were observed after drug inclusion into solid lipid nanoparticles. The liposomes formulations were investigated mostly for inhalation and injection administrations. Positive results were also observed. Authors underline the viability of incorporation of antituberculous drugs into phospholipid nanoparticles that may increase intestinal absorption and bioavailability. It is confirmed by authors' own data that showed increase of rifampicin efficiency after their incorporation into such nanoparticles.

[Drug delivery system on the base of phospholipid nanoparticles for rifampicin].

Low bioavailability of rifampicin, one of the main antituberculous drug, stimulates searches of its new optimized formulations. The present study has showen possibility of rifampicin embedding into nanoparticles from plant phosphatidylcholine (diameter of 20-30 nm). Addition of sodium oleate to the phospholipid system caused a 2-fold increase of the percent of rifampicin incorporation. After oral administration to rats, the maximal drug observed in plasma one hour after was 0.5 and 4.2 mkg/ml for free rifampicin for rifampicin in phospholipids-oleate nanoparticles, respectively. These levels were maintained for more than two hours of the experiment. High rifampicin bioavailability in the oleate containing phospholipid nanosystem suggests prospectivity of its pharmaceutical elaboration.