Thiacalixarene derivatives as agent for dna delivery into cells

FIELD: chemistry.

SUBSTANCE: invention refers to thiacalixarene derivatives of general formula I , which can be used as agents for DNA delivery into eukaryotic cells.

EFFECT: there are produced new heterocyclic compounds possessing the effective biological properties.

2 dwg, 8 ex

 

The invention relates to a novel series of chemical compounds, namely to derive thiacalixarene General formula I, which can be in configurations cone and 1,3-alternate:

ensure effective delivery of plasmid DNA into eukaryotic cells, which can find application in medicine as drugs for gene therapy, as well as transfection reagents for laboratory research.

In the scientific literature and patents, the date of submission of the present application, there are no examples of the use of derivative thiacalixarenes containing guanidinium groups on the lower rim of the macrocycle to transfer DNA into cells.

Studied the prior art, the applicant identified the most similar in structure and purpose the analogues of the claimed compounds representing derivatives of calix[4]arena and calix[6]arena containing fragments of arginine and guanidine connected to the macrocyclic platform through the corresponding hydrocarbon bridge (known in foreign literature, the term "spacer") [WO 2013021355].

The disadvantage of these compounds is the use as the basis of the molecular platform of calixarene, significantly limiting the number of possible configurational isomers obtained with�of dinani [N. Morohashi, F. Narumi, N. Iki, T. Hattori, S. Miyano, Chem. Rev., 2006, 106, 5291-5316.], the lack of data on the possibility of holding transfection of cell cultures, human NECT and higher cytotoxicity in comparison with the claimed compounds (according to data from the specified source concentration of compound which kills 50% of cells was 20 µmol/l, which is 23% lower than for the inventive compound (2)).

The applicant identified structural analogs of compounds (I) to the alkylated and their nano thiacalixarene of the General formula II, where D is S, CH2, SO, SO2group

E-H, alkyl, aryl, NO2NH2, NR2, guanidinium group, R-C=O, and In - group OR, where R is a hydrocarbon radical from C1to p10containing the triazole fragment, coupled with a carbohydrate residue. These compounds prevent the development of bacterial infections, however, they studied their transfection properties [US 2013252910 (A1)].

A disadvantage of the known compounds is that guanidinium group in compounds of the General formula II are on the "upper rim of the macrocycle in conjugation with aromatic fragments, which increases the acidity guanidinium fragment [Albert, A. The strength of heterocyclic bases / A. Albert, R. Goldacre, J. Phillips // J. Chem. Soc. - 1948. - P. 2240-2249], thereby reducing the efficiency of interaction with polyanions�mi DNA substrates. From the investigated prior art did not reveal the presence of their properties transfectional activity against eukaryotic cells.

Studied the prior art also known analogue of the claimed compounds for the purpose of constituting the polymer polyethyleneimine (PEI), providing efficient delivery of DNA into a variety of cell types [US 2012052574 (A1)].

A disadvantage of the known from the prior art polymer polyethyleneimine is its high cytotoxicity [Neu, M. Recent advances in rational gene transfer vector design based on poly(ethylene imine) and its derivatives / M. Neu, D. Fischer, Th. Kissel // J Gene Med. - 2005. - V. 7. - P. 992-1009] (0.009 mg/ml against 0.013 mg/ml and 0.050 mg/ml for the inventive compounds (1) and (2)).

Thus, studied by the applicant of the level of technology at the date of filing of application materials, the presence of close analogues both in composition and achieved the claimed technical solution to the technical(it) result(s).

The objective of the claimed technical solution is the creation of new effective, low-toxic drugs for effective delivery of DNA to eukaryotic cells.

The technical result of the new derivatives are thiacalixarene of formula (I), having transfectional activity against eukaryotic cells.

The claimed technical result is achieved by procedureswhich thiacalixarene of formula (I), having transfectional activity against eukaryotic cells.

The problem is solved by the synthesis of new compounds derived thiacalixarenes containing guanidinium group on the lower rim of the macrocycle in the configurations of the cone and 1,3-alternate formulas 1 and 2, which can be used as a means of delivery of DNA into eukaryotic cells.

The claimed compounds obtained by the interaction of the respective amine-based thiacalixarene with different guanidinium reagents.

The method is illustrated by the following examples, but is not limited to them.

Example 1. A two-stage method of obtaining 5,11,17,23-Tetra-tert-butyl-25,26,27,28-tetrakis-[3'-guanidinopropionic]tetracyclics[4]arena tetrachloride (1) by interaction of initial amine with N,N'-di(tert-butoxycarbonyl)-N"-triplescreen with subsequent removal of the protective tert-butoxycarbonyl of strong acid groups.

Stage 1. To a solution of 1.00 g of 5,11,17,23-Tetra-tert-butyl-25,26,27,28-tetrakis-(3-aminopropoxy)tetracyclics[4]arena in the configuration of the cone obtained according to literature methods [Galukhin, A. V. Mono-, 1,3-Di - and Tetrasubstituted p-tert-Butylthiacalix[4]arenes Containing Phthalimide Groups: Synthesis and Functionalization with Ester, Amide, Hydrazide and Amino Groups [Text] / A. V. Galukhin, N. E. Zaikov, I. S. Antipin, A. I. Konovalov, I. I. Stoikov // Macroheterocycles. - 2012. V. 5(3). - P. 266-274] in 40 ml of methylene chloride was dropwise added to dissolve� equimolar amount of N,N'-di(tert-butoxycarbonyl)-N"-trilingually in 10 ml of methylene chloride. Reaction mass was maintained for 24 hours. Then reaction mass was mixed twice with 30 ml of 2M aqueous solution of dehydrocholate sodium and 50 ml of saturated aqueous sodium bicarbonate solution, the organic phase was separated and dried over molecular sieves 3Å. Then, the solution was evaporated on a rotary evaporator, the residue was dried in vacuo over phosphorus oxide. The output of 5,11,17,23-Tetra-tert-butyl-25,26,27,28-tetrakis[3-(bis-tert-butoxycarbonyl-guanidine)propoxy]tetracyclics[4]arene (cone) amounted to 1.98 g (98%).

So a MP 248-250°C. the NMR Spectrum1H (CDCl3d, M. D., J/Hz):1.10 (D, s, (CH3)3C), 1.48 (D, s, Boc), 1.48 (D, s, Boc), 2.25 (6H, m,CH2CH_2CH23.67 (8H, m,NCH_2CH2CH2), 4.26 (8H, t,3JHH=6.9 Hz,OCH_2CHC2H2 ), 7.29 (8H, s, ArH), 8.46 (4H, t,3JHH=5.3 Hz,HN_CH2CH2CH2), 11.50 (4H, s,NH_Boc). The NMR spectrum13C (CDCl3d, M. D.), δ: 28.1, 28.4, 29.7, 31.2, 34.1, 38.1, 73.4, 78.9, 82.7, 130.0, 134.2, 146.1, 153.1, 156.2, 158.8, 163.7. The mass spectrum (MALDI-TOF): calculated [M+] m/z=1918.0, found [M+Na]+m/z=1941.6. Elemental analysis. Found (Percent):C, 58.81; H, 7.73; N, 9.12; S, 6.14. C96H148N12O20S4. Calculated (%): C, 60.10; H, 7.78; N, 8.78; S, 6.69. IR spectrum (liquid paraffin oil, ν/cm-1): 1274 (COC); 1636 (N-CO); 1723 (C=N), 3334 (NH, NH2).

Stage 2. To a solution of 0.50 g of 5,11,17,23-Tetra-tert-butyl-25,26,27,28-tetrakis[3-(bis-tert-butoxycarbonyl-guanidine)propoxy]tetracyclics[4]arene (cone) in THF was added 2 ml of concentrated hydrochloric acid. Reaction mass was maintained for 24 hours. Further, the reaction mass is maximum was evaporated on a rotary evaporator, was added 50 ml of diethyl ether. Precipitated white precipitate was filtered, washed with two portions of diethyl ether and 30 ml then dried in a desiccator over phosphorus oxide. The yield of the target Conn�tion (1) was 83%.

Div.>238°C. the NMR Spectrum1H (D2O, δ, M. D., J/Hz): 0.98 (D, nl.C, (CH3)3(C), 2.15 (6H, br.m,CH2CH_2CH2), 3.46 (8H, br.t,NCH_2CH2CH2), 4.22 (8H, br.t,OCH_2CHC2H2), 7.30 (8H, br.s, ArH). The NMR spectrum13C (125 MHz, CDCl3), δ:29.2, 30.8, 33.8, 38.1, 73.1, 129.4, 134.1, 146.0, 157.1, 158.4. The mass spectrum (MALDI-TOF): calculated [M+] m/z=1262.5, found [M-4HCl]+m/z=1117.4. Elemental analysis. Found (Percent):C, 47.92; H, 5.63; N, 10.42; S, 7.89. C56H88N12O4S4Cl4. Calculated (%): C, 48.81; H, 5.70; N, 10.67; S, 8.15. IR spectrum (liquid paraffin oil, ν/cm-1):1265 (COC); 1662 (NH2); 3176 (=NH).

Example 2. Three-stage method of obtaining 5,11,17,23-Tetra-tert-butyl-25,26,27,28-tetrakis(3'-N,N-bis[3-aminopropyl]guanidinopropionic) tetracyclics[4]arena Oct�lorida (2).

Stage 1. In a round-bottom flask equipped with a magnetic stirrer and reflux condenser with chlorellas tube, a mixture of 0.50 g (0.53 mmol) of 5,11,17,23-Tetra-tert-butyl-25,26,27,28-tetrakis(3-aminopropoxy)-tetracyclics[4] arena 1,3-alternate, obtained according to literature methods [Galukhin, A. V. Mono-, 1,3-Di - and Tetrasubstituted p-tert-Butylthiacalix[4]arenes Containing Phthalimide Groups: Synthesis and Functionalization with Ester, Amide, Hydrazide and Amino Groups [Text] / A. V. Galukhin, E. N. Zaikov, I. S. Antipin, A. I. Konovalov, I. I. Stoikov // Macroheterocycles. - 2012. V. 5(3). - P. 266-274], 1.41 g (5.27 mmol) of N-(3-bromopropyl)phthalimide, 0.72 g (5.27 mmol) svejeporublennogo potassium carbonate and 40 ml of freshly distilled acetonitrile was held at reflux for 40 hours. After cooling the reaction mixture, the potassium carbonate was filtered, washed with chloroform, 2×20 ml the resulting solution was evaporated under reduced pressure, the residue is recrystallized from methyl alcohol. The output of 5,11,17,23-Tetra-tert-butyl-25,26,27,28-tetrakis-(3-N,N-di[3-phthalimidopropyl]aminopropoxy)tetracyclics[4]arena amounted to 0.95 g (74%).

So a MP 254°C. the NMR Spectrum1H (CDCl3d, M. D., J/Hz): 1.22 (D, s, (CH3)3C), 1.82 (24H, m,CH2CH_2CH2), 2.52 (24H, m, N-CH2), 3.73 (S, t,3JHH=7.4 Hz, CH2-Pht), 3.95 (8H, �, 3JHH=7.6 Hz, O-CH2), 7.40 (8H, s, Ar1-H), 7.63-7.70 (8H, m, Ar2-H), 7.75-7.82 (8H, m, Ar2-N). The NMR spectrum13With (CDCl3d, M. D.), δ: 27.70, 28.69, 32.78, 35.47, 37.75, 51.54, 52.20, 71.25, 123.83, 129.23, 132.75, 132.90, 134.34, 145.22, 159.16, 168.64. Spectrum of1N1H NOESY (NOE) (the most important cross-peaks): H4b/N7, H4b/N9, H3/N7. IR spectrum (liquid paraffin oil, ν/cm-1): 1262 (COC); 1706, 1772 (C=O). The mass spectrum (MALDI-TOF): calculated [M+] m/z=2446.0, found [M+H]+m/z=2447.0. Elemental analysis. Found (Percent): C, 69.01; H, 6.66; N, 7.36; S, 4.16. C140H148N12O20S4. Calculated (%): C, At 68.72; H, 6.10; N, 6.87; S, 5.24.

Stage 2. In a round-bottom flask equipped with a magnetic stirrer and reflux condenser with chlorellas tube, 1.00 g of 5,11,17,23-Tetra-tert-butyl-25,26,27,28-tetrakis-(3-N,N-di[3-phthalimidopropyl]aminopropoxy) tetracyclics[4]arena and 2 ml of hydrazine hydrate (40 mmol) in a mixture of 30 ml of ethanol and 20 ml of tetrahydrofuran was boiled for 5 hours. After cooling the reaction mixture the solvent was evaporated in vacuo, poured 50 ml of CHCl3and washed the organic layer with 20 ml of 20% NH4OH. The organic phase was separated and dried over molecular sieves 3Å. The solvent was distilled under reduced pressure. The output of 5,11,17,23-Tetra-tert-butyl-25,26,27,28-tetrakis(3'-N,N-bis[3-aminopropyl]aminopropoxy) tetracyclics[4]the arena was 89%.

Etc�. 137-139°C. the NMR Spectrum1H (CDCl3d, M. D., J/Hz): 1.23 (D, s, (CH3)3C), 1.38 (D, nl.with NH2), 1.59 (8H, m,CH2CH_2CH2), 1.80 (M, m,CH2CH_2CH2), 2.46 (24H, t,3JHH=7.1 Hz, N-CH2), 2.73 (N, t,3JHH=6.9 Hz, CH2-N), 4.00 (8H, t,3JHH=7.9 Hz, O-CH2), 7.40 (8H, s, Ar1-N). The NMR spectrum13C (CDCl3d, M. D.), δ: 31.4, 31.5, 33.8, 34.6, 36.2, 40.3, 41.8, 67.6, 128.2, 128.6, 145.5, 158.0. Spectrum of1H-1H NOESY (NOE) (the most important cross-peaks): H4b/N7, H3/N7. IR spectrum (liquid paraffin oil, ν/cm-1): 1266 (COC); 3362 (NH2). The mass spectrum (MALDI-TOF): calculated [M+] m/z=1392.9, found [M+H]+m/z=1394.1. Elemental analysis. Found (Percent): C, 64.68; H, 9.37; N, 12.04; S, 8.75. C75H132N12O4S4. Calculated (%): C, 64.61; H, 9.54; N, 12.06; S, 9.20.

Stage 3. In a round-bottom flask equipped with a magnetic stirrer, 0.30 g (0.21 mmol) of 5,11,17,23-Tetra-tert-butyl-25,26,27,28-tetrakis(3'-N,N-di-[3-aminopropyl]am�nephropexy)tetracyclics[4]arene and 0.30 g (2 mmol) of S-allylisothiocyanate chloride in 20 ml of tetrahydrofuran was boiled for 20 hours. After cooling, the reaction mixture was filtered and the precipitate was washed with 3×20 ml of tetrahydrofuran. The precipitate was dried in a desiccator over phosphorus oxide. The target compound (2) was obtained with a yield of 93%.

5,11,17,23-Tetra-tert-butyl-25,26,27,28-tetrakis(3'-N,N-bis[3"-guanidinopropionic]aminopropoxy)tetracyclics[4]arene octaploid (2). Div.>208°C. the NMR Spectrum1H (D2O, δ, M. D., J/Hz): 1.19 (D, s, (CH3)3C), 1.79 (24H, m,CH2CH_2CH2), 2.55 (24H, nl.t, N-CH2), 2.93 (M, nl.t,CH_2NH), 3.13 (8H, br.t,OCH2CH2CH_2N) 3.90 (8H, br.t, O-CH2), 7.52 (8H, s, Ar1-H). The NMR spectrum13C (125 MHz, D2O), δ: 23.7, 25.8, 30.6, 33.8, 38.0, 39.3, 39.4, 49.2, 49.8, 128.6, 132.2, 146.3, 156.8. Spectrum of1H-1H NOESY (NOE) (the most important cross-peaks): H4b/N7 , H4b/N9, H4b/N12. The mass spectrum (MALDI-TOF): calculated [M+] m/z=2032.9, found [M-8HCl]+m/z=1741.1. Elemental analysis. Found (Percent): C, 48.92; H, 7.33; N, 19.22; S, 6.19. C84H156N28O4S4Cl8. Calculated (%): 49.60; H, 7.73; N, 19.28; S, 6.31. IR spectrum (liquid paraffin oil, ν/cm-1): 1267 (COC); 1663 (NH2); 3175 (=NH).

To describe the properties thiacalixarenes, which is the subject of the invention, representative data obtained for compounds (1) and (2) differing in the length of the GS spacer fragment connecting guanidinium group with macrocyclic platform, the number of cationic groups and the configuration of the macrocyclic platform (cone (1) and 1,3-alternate (2)).

Abbreviations

PBS - phosphate buffered saline

eGFP green fluorescent protein

DMEM - modified Dulbecco medium eagle. Nutrient medium for cell cultures

Medium with serum - DMEM and 10% of the volume of serum of bovine embryos (HyClone)

MTT 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide

CD50is the concentration at which killed 50% of cells in culture.

Example 3. The methodology of experimental transfection using new compounds (1) and (2)

Monoclonal culture of human cell lines NECT seated on 40 thousand cells per well the day before transfection based to get on the trail�schy day confluently 50-70%. For transfection NEXT use concentration of 50-100 thousand cells per well of 96-hole tablet. One hour before transfection, the cells give a fresh DMEM medium without serum and antibiotics. Transfection is carried out in 96-well plate, experiments were carried out in the triple repetition. First make a series of dilutions of potential transfectants in water Milli-Q with a final concentration of 10-3, 10-2, 0.1 and 1 mg/ml, was added to the DNA solution and gently pipetimout once. After 10-15 minutes of incubation liposonix with 0.6 μg of plasmid DNA peGFP-N1 (Clontech) in a total volume of 20 µl was added to cells without significant mixing. The following day, the fluorescence of the cell population in the tablet see the fluorescent microscope with inverted optics. 48 hours after transfection analyze by flow cytometry as described below. After receiving the results of the preliminary functionality of the new compounds as transfectants, conduct further optimization of the ratio of the number of transfectants to the amount of plasmid DNA near that level at which occurs the maximum transfection efficiency. Used step in the selected range is a serial two-fold increase in concentration. Also optimize the duration of incubation of the cells with battery�klonoa mixture and the possibility of combining transfection/electroporation. Conduct after electroporation transfection is inappropriate, because it causes cell death, significantly without affecting the percentage of transfected cells. The optimal time of incubation with transfectants is 4 hours after which the medium in the cells change to full DMEM with serum and antibiotic.

Example 4. The method of analysis transfection of cells by flow cytometry

In the study the degree of transfection of cells using the method of flow cytometry. Each well pad with cells after incubation with experimental drugs within 48 hours, washed with 200 µl of PBS (x1), treated with trypsin and fixed in 0.5% formalin. In the thus obtained cell suspension determined by flow cytometry the percentage of cells expressing eGFP. This value is a measure of the efficiency of transfection. Cells incubated without experimental drugs under the same conditions, using as negative control. For comparison, a measurement of the percentage of eGFP positive cells, transfected using a commercial preparation Metafectene easy on the Protocol proposed by the manufacturer (Biontex, EU). Further, from the data obtained calculate the percentage of fluorescent cells using FACS Diva (Becton Dickinson, USA), averages and standard�tnye deviations for the populations.

Example 5. The method of determining the cytotoxicity of the compounds to the cell line NEKT

Cytotoxicity was determined using MTT reagent. As a criterion of cytotoxicity option was used CD50(concentration at which killed 50% of cells in culture). Culture of human cell lines NECT seated on 40 thousand cells per well of 96-well plates the day before the determination of cytotoxicity of the calculation to obtain the next day, 50-70% of a monolayer. An hour before the addition of experimental compounds or their complexes with DNA cells give fresh DMEM without serum and antibiotics. Make a series of dilutions of potential transfectants in water Milli-Q with a final concentration of 10-3, 10-2, 0.1 and 1 mg/ml, is added to a solution of plasmid DNA and gently pipetimout once. After 15 minutes incubation liposonix with 0.6 µg of plasmid DNA in a total volume of 20 µl was added to cells without significant mixing. In parallel to the cells add individual substances without DNA in an appropriate amount. After 48 hours of incubation with the compounds to the samples was added to 20 µl of R-RA MTT (5 mg/ml). After 2-4 hours of incubation with MTT remove the medium from the wells, taking care not to touch the crystals, dissolve the formazan crystals in 200 μl DMSO. Next, determine the optical density of slip casting�ü the instrument Tecan Sunrise Basic at a wavelength of 492 nm with subtraction of background. The data obtained was determined parameter CD50for the compounds of formulas 1 and 2, which are represented in the diagram of Fig.1, which shows cytotoxicity (CD50, µmol/l) compounds (1) and (2) to the cell line NECT.

From this diagram it follows that the value of cytotoxicity (CD50) the compounds (1) and (2) decreases when their complexation with plasmid DNA, and also shows that the concentration range of values of CD50for compounds (1) and (2) and their complexes with plasmid DNA higher than the effective concentrations of these compounds in which there is the highest percentage of transfected cells (see Example 7 and 8), which allows to use the data connection for the claimed purpose.

Example 6. The complexes of the compounds with DNA

When increasing concentrations of potential transfectants until N/P (the ratio of the number of positively charged nitrogen atoms of transfectant to the number of phosphate residues in plasmid DNA) of about 1 (for compounds (1) and 4 (compound (2)) shows almost complete disappearance of free plasmid DNA, which is an indication of complex formation. In photographs a and b (Fig. 2) shows the results of electrophoresis, plasmid DNA (4700 PN) in the presence of increasing concentrations thiacalixarene (1) and (2) N/P=0 �about N/P=64, demonstrating the decrease in the proportion of free plasmid DNA in the increase of the ratio N/P is from 0 to 64.

Thus, the data for 1 and 2 it follows that the fraction of free plasmid DNA in the increase of the ratio N/P is from 0 to 64 decreases.

Example 7. Conducting transfection using the compound (1)

Under the above Protocol testing transfectional the effectiveness of the compounds (1). The connection (1) the highest percentage of transfected cells is 42% (average of 30±10%) at a concentration of 11.1 mmol/L.

Example 8. Conducting transfection using compound (2)

Under the above Protocol testing transfectional the effectiveness of compound 2. The highest percentage of transfected cells is 28% (average 20±8%) at a concentration of 6.9 mmol/L.

Thus, the claimed derivatives thiacalixarene 1 and 2 possess the ability to effectively deliver DNA into eukaryotic cells and have low toxicity. The obtained results represent the first example of a successful transfection of eukaryotic cells derived thiacalixarene.

The claimed technical solution meets the criterion of "novelty", presented to the invention, because of the investigated prior art is not identified technical solutions, characterized provide during account creation�mi signs leading to achieve the stated technical results of the claimed technical solutions.

The claimed technical solution meets the criterion of "inventive step", presented to the invention, since it is not obvious to those skilled in the art, due to the fact that the claimed technical solution provides the implementation of objectively existing contradictions in practice, not solvable by conventional design, namely of the investigated prior art it is known that the existing system of delivery of DNA into cells on the basis of demokraticheskih derivatives have a high cytotoxicity, which prevents their use for the stated purpose, wherein said cytotoxicity was significantly reduced by the use of macrocyclic platform thiacalixarene, also providing the spatial organization of binding guanidinium fragments, thereby creating an effective transfection agents. Stated macrocyclic compounds exhibit low cytotoxicity, and the ability to carry out the transfection of the cell line NEXT, thus it can be concluded that the obtained results is not obvious to a person skilled in the art.

The claimed technical solved�e meets the criterion of "industrial applicability", because they can be implemented on any enterprise using standard equipment, known materials and technologies.

Derivatives thiacalixarenes formula (1) and (2),

as a means of delivering DNA into eukaryotic cells.



 

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3 tbl

FIELD: medicine.

SUBSTANCE: invention refers to medicine and can be used for making a drug for removing subcutaneous fat. That is ensured by using a composition containing: at least one phospholipid, at least one glycyrrhizic acid or a glycyrrhizic acid salt, and wherein total content of phospholipids and glycyrrhizic acid or its salts makes 2-80 wt % and a weight ratio of phospholipids and glycyrrhizic acid or its salts makes from 30:1 to 0.5:1. The composition can additionally contain additives. As phospholipid, the composition contains animal or herbal phosphatidylcholine. The composition can also contain glycyrrhizic acid or potassium, sodium, ammonium or magnesium salt of glycyrrhizic acid. As an additive, the composition contains sugar, particularly glucose, or maltose, and/or their derivatives, mannitol, sorbitol or lactose. The composition contains phosphatidylcholine in a total amount of 15 to 98 wt %, preferentially 30 to 98 wt %, more preferentially 50 to 98 wt %, especially preferentially 75 to 98 wt %, and most preferentially 75 to 90 wt % of total content of phospholipids. The composition is used in the dry form, preferentially in the form of lyophilisate prepared by freezing and drying, or in the form of a solution. The composition can contain physiologically acceptable solvents, including water, normal saline, glucose solution, such monohydric alcohols, as ethanol, 2-propanol, n-propanol, such polyatomic alcohols, as glycerol and/or propane diol, such polyglycols, as polyethylene glycol and/or Miglyol, glycerol, formal, dimethylisosorbitol, natural and synthetic oils and/or esters. Diseases of subcutaneous fatty tissue can be particularly related to local fat maldistribution. Fatty tissue tumour decomposition and reduction can be also solved. The drug can be presented in the form of cream, ointment, gel, hydrogel, lotion, paste, powder or solution. Undesired unaesthetic and pathological local fat maldistribution involve lipoedema, adipose growth, abdominal adiposis, cellulites, pseudogynecomasty, "buffalo hump" in HIV-infected patients, panniculitis or nonspecific subcutaneous fat deposition. The preparation is administered by subcutaneous, intra-abdominal, intramuscular or intravenous injection. The administration is implemented by a method specified in a group consisting of ioophoresis, electroporetion and phonophoresis.

EFFECT: described invention is successfully used for the above purposes.

17 cl, 11 ex, 7 tbl, 11 dwg

FIELD: medicine.

SUBSTANCE: what is presented is using meso-tetra(3-pyridyl)bacteriochlorin of structural formula (I) as a near-infrared photosensitiser for a photodynamic therapy. Doses 1.0-2.5 mg/kg of the declared photosensitiser have provided 70-100% tumour growth inhibition, 80-131% increase in life expectancy and 25-100% animals' recovery by selective tumour collection and fast clearance.

EFFECT: high photoinduced activity on human tumour cells of various epithelial origin and high dose-dependent anti-tumour effectiveness in the animal with tumours of various origins.

5 dwg, 8 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: presented invention refers to immunology. There are disclosed versions of a dimer compound for forming a multimer capable to reproduce the effector function of aggregated IgG with identical monomers. Each monomer of the dimer comprises: a monomer of IgG2 link region or a monomer of isoleucine zipper, dimerising each of which forms a multimerising region, and at least Fc-domain monomer containing a link region, CH2 domain and CH3 domain of IgG1. What is described is a multimer compound capable to reproduce the effector function of aggregated IgG and containing two or more dimers. There are disclosed a method for changing the immune response using the dimer or multimer, as well as a multimer-based method of treating an inflammatory disease.

EFFECT: using the invention provides the new compounds capable to bind at least one FcR specified in a group consisting of: FcγRI, FcγRII, FcγRIII, FcγRIV, or their non-human version that can find application in medicine for IVIG substitution for treating a wide range of diseases, including the inflammatory and autoimmune diseases.

7 cl, 25 dwg, 5 tbl, 25 ex

FIELD: veterinary medicine.

SUBSTANCE: product comprises Lycopodium clavatum, Acidum arsenicosum, Phosphorus, Podophyllum peltatum, Thuja occidentalis, Echinacea purpurea, Silybum marianum, Selenocysteine, and the components are taken in the dilutions described below in the following ratio, in parts: Lycopodium clavatum ⌀=D1 0.004, Podophyllum peltatum ⌀ 0.003, Acidum arsenicosum ⌀=D2 0.0001, Phosphorus ⌀=D3 0.001, Thuja occidentalis ⌀ 30, Echinacea purpurea ⌀ 30, Silybum marianum ⌀ 60, Selenocysteine 0.2.

EFFECT: product has an effective stress-protective and growth-stimulating effect, it regulates the metabolism in young farm animals.

3 cl, 10 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: method for producing an agent for stimulating body cells involving preparing a mixture of aqueous solution of selenious acid and PEG 400; that is followed by preparing a mixture of hydrazine hydrochloride and PEG 400; the prepared mixtures are combined; the solution is put to dialyse against distilled water; surplus of water is driven off; the produced solution is added with hexamethylene tetramine; pH is reduced to 7.2-7.4; the method is implemented in certain circumstances.

EFFECT: producing high-effective, ecologically safe agent by the synergism of colloidal selenium and hexamethylene tetramine on body cell stimulation.

1 dwg, 2 tbl, 4 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine and can be used for the treatment of radiation-thermal injury of an organism. For this purpose a single subcutaneous introduction of bifidumbacterin, irradiated by gamma-rays in a dose of 14.0 Gy, is carried out. Bifidumbacterin is introduced in a dose of 1.43·106 CFU/kg. After that, 10% hypericum oil is applied on the burnt region. Then a 10% hypericum cream is applied after 3-4 days.

EFFECT: method makes it possible to carry out the treatment of combined radiation-thermal injuries in an effective way with the application of available and cheap pharmacotherapeutic means.

1 tbl, 4 ex

FIELD: medicine.

SUBSTANCE: erythrocyte cell medium is added with an aqueous solution of sodium and potassium salts of humic acids prepared on brown coal of leonardite in a dose of 10.0 mg/kg. That is incubated at a temperature of 37°C for 40 minutes before treatment with acidic haemolytic.

EFFECT: invention enables normalising the cell membrane permeability and reducing the damaged cell count under acidic haemolytic.

1 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to new compounds of formula I, wherein R1 and R2 are identical or different and specified in an alkyl or alkenyl hydrocarbon chain; the R3 group values split by lipase are specified in the patient claim. R4 and R5 are independently hydrogen or C1-C7alkyl; R6 represents hydrogen or C1-C7alkyl; and R7 and R8 are independently hydrogen or C1-C7alkyl. The invention also refers to using compounds of formulas ,

which are introduced into the mammalian biological system and increase the cell concentrations of specific sn-2 substituted ethanolamine-plasmalogens.

EFFECT: compounds are applicable in treating or preventing the age-related disorders associated with high membrane cholesterol, high amyloids and low plasmalogens, such as neurodegeneration, cognitive disorder, dementia, cancer, osteoporosis, bipolar disorder and vascular diseases.

11 cl, 18 dwg, 7 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to compositions of an antioxidant composition aimed at the suppression of oxidative stress in type 2 diabetes. The said compositions contain, counted per 1 dose: 50-120 mg of coenzyme Q10, 30-160 mg of dihydroquercetin, and 30-60 mg of A-lipoic acid or 50-100 mg of coenzyme Q10, 50-100 mg of dihydroquercetin, 30-60 mg of A-lipoic acid and 50-100 mg of nicotineamide.

EFFECT: compositions possess an antioxidant activity and prevent the development of the fatty tissue dysfunction.

2 cl, 1 dwg, 2 tbl, 2 ex

FIELD: biotechnology.

SUBSTANCE: invention relates to glycosidic derivatives of 1,2-dithiole-3-thione or 1,2-dithiol-3-one of formula 1 , where R1=S or O; R2 is a residue of per-O-acetyl D-glucose, per-O-acetyl D-galactose, per-O-acetyl D-mannose, per-O-acetyl D-xylose, per-O-acetyl L-arabinose, per-O-acetyl-D-maltose or D-glucose, which can be used against cancerous diseases.

EFFECT: new bioactive compounds with cancerous preventive action and pharmaceutical products based on them are proposed, which display the cancerous preventive effect in non-cytotoxic concentrations.

2 cl, 4 ex, 5 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions refers to medicine and concerns a local topical composition characterised by the fact that it contains synthetic lipoic acid; 95% ethyl alcohol; 1,2-propylene glycol, polyacrylic acid (any other hydrophilic gelling agent), triethanolamine, purified water; a method for preparing the local topical composition characterised by the fact that synthetic lipoic acid is dissolved while stirring in mixed ethyl alcohol and propylene glycol; at the same time polyacrylic acid is dispersed in a specified amount of water sufficient to reduce the ratio of the ingredients up to 100 wt %, after 3-hour swelling, the prepared solution is added to synthetic lipoic acid dissolved in mixed ethyl alcohol and propylene glycol and mixed up; the prepared composition is neutralised by a triethanolamine solution to pH 5.5-6.5 and mixed to form a light-yellow transparent gel-like mass.

EFFECT: group of inventions provides high antirheumatic, anti-inflammatory and analgesic activity, as well as effective antibacterial and anti-burn action.

4 cl, 4 ex, 1 tbl, 3 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: composition for treating oxidative stress comprises ball-shaped lipoic acid or one of salts thereof, and at least one lipophilic medium. The lipoic acid balls represent particles consisting of an inert core (a nucleus) coated with lipoic acid which is coated with a first layer of an isolating polymer, and with a second polymer layer resistant (stable) at gastric pH. What is also described is a preparation for treating oxidative stress with an unified dose containing the above composition. The preparation is presented in the form of a soft gelatin capsule.

EFFECT: compositions according to the invention are stable in the lipophilic medium.

22 cl, 15 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to surgery, and may be used for prevention of acute postoperative pancreatitis. For this purpose, the underlying background therapy is combined with intravenous bolus administration of dalargin 0.002 g and the antioxidant thioctic acid depending on a risk level of the complication measured in terms of the area of intervention in abdominal operations. A high risk level requires thioctic acid to be administered in a dose of 600 mg intravenously drop-by-drop 1 hour before the operation and on the following day, and thioctic acid in a dose of 300 mg on the third day intravenously drop-by-drop. In a moderate risk level thioctic acid is administered intravenously drop-by-drop in a dose of 600 mg 1 hour before the operation and in a dose of 300 mg on the following day. And in a low risk level, thioctic acid is administered intravenously drop-by-drop in a dose of 300 mg 1 hour before the operation.

EFFECT: method enables the more effective prevention of acute postoperative pancreatitis ensured by the combined use of drugs with antisecretory and antioxidant activity.

2 tbl, 3 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutics and medicine, and concerns a pharmaceutical composition for preventing and treating diseases mediated by high expression or high activity of liver X receptor (LXRa), high expression or high activity of a sterol regulatory element-binding protein-1c (SREBP-1).

EFFECT: preventing and treating renin hypertension, hyperaldosteronism, adrenoleukodystrophy, glomerular sclerosis, proteinuria, nephropathy, hepatic steatosis, hypertriglyceridemia or hyperreninemia, possessing high activity.

6 cl, 11 dwg, 9 ex

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