Hydrated n-fullerene-amino acid derivatives, method for preparing them and based pharmaceutical compositions

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to new hydrated N-fullerene-amino acids of general formula C60(H)3{NH(CH2)nCOOH}3·xH2O, wherein C60 represents fullerene, n = 5-7, x = 8-10, which possess herpes virus, influenza virus, HIV activity, as well as anticancer and antipsoriatic activity. The invention refers to a method for preparing said fullerene amino acids and based pharmaceutical compositions.

EFFECT: preparing new hydrated N-fullerene-amino acids.

5 cl, 28 tbl, 10 ex

 

The invention relates to pharmaceutical industry and medicine and relates to a new hydrated amino acid derivatives of fullerene C60formula (I), and the method of their derivation and creation of pharmaceutical compositions based on them.

The use of fullerenes as biologically active compounds has led to an intensive development of the chemistry of functional derivatives of fullerene, especially after it was shown that the number of water-soluble fullerene derivatives show high antiviral activity (Partha R, Conyers JL, "Biomedical applications of functionalized fullerene-based nanomaterials" Int. J. Nanomedicine, 2009, 4, 261-75 Pat US 6204391, 2005, "Water soluble fullerenes with antiviral activity", R.Bakry et al., "Medicinal application of fullerenes" International Journal of Nanomedecine, 2007 (4) 639-649, Z.Zhu, D.I.Schuster, M.Tuckermann, "Molecular Dynamics Study of the Connection between Flap Closing and Binding of Fulleren-Based Inhibitors of the HIV-1 Protease", Biochemistry, 2003, v.42, 1326-1333).

The use of fullerene derivatives in medicine is based on the lipophilic properties of fullerene core, allowing fullerene derivative to penetrate through cell membranes, and the ability of the fullerene with high quantum yield to generate singlet oxygen that breaks down DNA. These properties provide a functional derivative of fullerene manifestation of cytotoxic, antiviral and other properties (D. Bedrov, G.D. Smith, Davande N., "Passive transport of fullerenes through a script membrane." J. Phys. Chem., B, 2008, v.112., p.2078-84, Qiao, R., Roberts, A.E., "Translocation of fullerene and its derivatives across a lipid bilayer", Nano Lett., 2007, v.7, p.614-9. Nelsen G.D., and others, "In vivo biology and toxicology of fullerenes and their derivatives", Basic and Clinical Pharmacology and Toxicology, 2008, v.103, p.197-208).

Hydrated forms of fullerene exhibit high biological activity as the bioantioxidants, due to the formation of active structural forms of water clusters, coordinated the field of fullerene (Andrievsky G.V., Brushkov V.I., Tykhonov A.A., Seth William page S.V. "Peculiarities of the antioxidant and radioprotective effects of hydrated C60fullerene nanostructures in vitro and in vivo". Free Radical Biology and Medicine, 2009, v.47, p.786-793).

The main problem impeding biological studies of fullerenes and their derivatives and the creation of therapeutic drugs based on them, is the difficulty of introducing fullerene systems in aqueous solutions.

A promising method of obtaining water-soluble fullerene compositions is chemical modification of fullerene sphere by introducing hydrophilic solubilizing ligands. At the present time has received a large number of functionalized fullerenes containing hydrophilic fragments in the side chain attached to the fullerene ligands (detergent type complexes), and spherical type derivatives when there are polar groups, distributed fullerene sphere (this type includes fullerenol, eminiaddict).

The most promising DL is using are amino acid derivatives of fullerene.

Unnatural amino acids are aliphatic series containing 6 or more methylene groups show a number of features that appear in the processes of hydration and their biochemical activity. Spectral studies of the structure of water in aqueous solutions of amino acids indicate that the increase in the number of methylene groups between the amino and carboxyl groups leads to increased degradation of water clusters. Studies of the pharmacological properties of derivatives of amino acids of a wide range of R-(CH)nCOOH showed higher activity systems with n is greater than and equal to 6.

Derivatives of fullerene C60with amino acids spherical type obtained by the reaction of nucleophilic attachment of amino acids on the amino group to the field of fullerene described in the patents of the Russian Federation No. 2196602, 2124022, 2236852 that you can offer as analogues of the present invention.

In the patent of Russian Federation №2196602 a method of inhibiting the reproduction of HIV and CMV infections using the compounds based on amino acid and dipeptide derivatives of fullerene. As the amino acid derivative of fullerene used sodium salt fullerene-aminocaproic fullerene-aminobutyric acid.

In the patent of Russian Federation №2124022 to obtain fullerene-aminocaproic acid to a solution of fullerene in o-dichlorobenzene add an aqueous solution of potassium salt aminaka ronojoy acid and 18-crown-6. The reaction mass is stirred for 6-8 hours at 60°C. Then the solvent is distilled off, the residue is treated with saturated solution of potassium chloride and the remainder of the fullerene derivative is washed with water. The yield of the desired product quantitatively. Received (monohydro)N-fullerene-aminocaproic acid is soluble in dimethylsulfoxide, dimethylformamide, pyridine. In the claimed method of synthesis is not defined conditions for the allocation of the final product.

The main disadvantage of the obtained compounds, which are products of monopolisation is their insolubility in water. Another disadvantage of the invention is the use in their synthesis of crown ether as a phase transfer catalyst, which is difficult to separate from the reaction products.

In the patent of Russian Federation №2236852 protected means for inhibiting reproduction enveloped viruses representing fullerenelike anions with the General formula C60Hn[NH(CH2)mC(O)O-]nresulting from the interaction of fullerene with the salt of the amino acids in the medium of organic solvent in the presence of polyalkylated.

To obtain these compounds to the solution of fullerene in o-dichlorobenzene (toluene or other organic solvent) contribute the amino acid in the form of salts (potassium or sodium), then to ablaut the solubilizer. The order of entering into reaction medium amino acids and a solubilizer is not important, you can make them as complex, pre-mixing. As a solubilizer use different polyalkyloxy: glycols mol. mass of from 150 to 400 above 400 (e.g., PEG-1500), and polyethylene glycols having a free end groups, but with substituted (for example, dimethyl ether of polyethylene glycol (mol. weight 500). To increase the rate of reaction add any strong reducing agent (alkali metals). The ratio of the fullerene and amino acids increased more than 50 times. The transformation into the desired pharmaceutically acceptable salt, especially sodium or potassium, was carried out by treating the acid with a suitable base or by adding a salt of the weak volatile acid. In particular, it is not soluble in water fullerenelike acid into the preferred pharmaceutically acceptable salts such as sodium salt, which is soluble in water. The salt of the weak volatile acid by treatment of a solution of a salt of an alkali metal and a weak volatile acid. When the concentration of the solution by evaporation or lyophilization of a weak acid is removed and the mixture fullerenelike acid is secreted in the form of mixtures of their alkali metal salts. Target product in this image is in the shadow is characterized by the constancy of the composition, the content of the target product of the basic substance is only of 87.8%.

The main disadvantages of fullerene-amino acid derivatives obtained are presented in patent method, is that this way we obtain a mixture tolerancerelated anions as salt and acid forms. To get a individual connection with a method described in the patent, is not possible. Also fullerenealuminum obtained by the patented process, in acid form, practically insoluble in water. To get a stable pharmaceutical composition with fulleropyrrolidine anions was not possible, because in the process of storage compounds precipitate. Fullerenealuminum influence leucopoiesis: cause a shift leukocyte and induce the emergence of new forms of neutrophil - neutrophil metamyelocytes in laboratory animals (rats and rabbits). From the point of view of security (safety) this indicates the presence of these substances toxicity that causes these changes. Application in the synthesis of a large excess of potassium or sodium salts of amino acids and a large excess of solvent leads to environmental problems associated with disposal of waste production and increase the cost of the production process. The use of school is full of metals to increase the rate of reaction is technologically impossible when using chlorinated aromatic solvents.

The objective of the proposed technical solution is to obtain an individual hydrated fullerene compounds With60with aminocarbonyl acids, possessing activity against herpes virus, hepatitis C virus, influenza viruses of different nature, HIV and antitumor and protivopsoriaticescoe activity that do not have a toxic effect on the organism; the method of obtaining these compounds and pharmaceutical compositions comprising these compounds.

To achieve the objectives of the proposed group of inventions combined to form a single inventive concept: the connection method thereof and pharmaceutical compositions containing the specified connection.

The problem is solved individually gidratirovannym connection fullerene C60with aminocarbonyl acids of General formula (II), characterized by the fact that one molecule of fullerene have three covalently linked amino acid fragment having the structure of the active centers of hydration, leading to the formation of water-soluble hydrates, and a long hydrocarbon chain that allows it to retain water molecules in the inner coordination sphere of fullerene complexes.

This task is solved in that the hydrated fullerene production is haunted amino acids of the formula (II) are formed by the interaction of fullerene with a 15-fold molar excess of anhydrous potassium salts of amino acids in the environment of aromatic organic solvent by slow addition to obtained slurry phase catalyst with stirring and heated to a temperature not higher than 60-80°C until complete discoloration of the solution and formation of a solid precipitate, which is then allocated, followed by the processing of 0.8 M aqueous solutions of potassium salts of fullerene-amino acid of 0.1 N solution of an organic or mineral acid, followed by centrifugation, washing and drying the precipitate.

Also according to the invention the anhydrous potassium salt of amino acids used in fine condition, which improves the reactivity of the process, its effectiveness and efficiency, and allocation of solid precipitate potassium salts of fullerene-amino acids is performed by filtering, washing with ethanol and drying. As phase transfer catalyst is used, the methyl ethers of polyethylene oxides of molecular weight 200, 400, 500 as the most accessible and secure catalysts.

This task is solved by the creation of the pharmaceutical compositions containing as active substance water-soluble hydrated fullerene-amino acids of formula (II)having activity against herpes virus, hepatitis C virus, influenza viruses of different nature, HIV and antitumor and protivopsoriaticescoe activity.

Pharmaceutical compositions for predlojeno is the technical solution containing the compound of General formula (II), effective to achieve the desired result, and can be entered as standard medicinal forms (for example, in solid, semisolid, or liquid form), containing compounds of the proposed technical solution as an active ingredient in a mixture with a carrier or excipient suitable for intramuscular, intravenous, oral, sublingual, inhalation, local, intranasal and intrarectal administration. The active ingredient can be included in a composition together with commonly used non-toxic pharmaceutically acceptable carriers suitable for the manufacture of solutions, tablets, pills, capsules, pills, suppositories, emulsions, suspensions, ointments, gels or any other medicines.

A particular level of dosage and frequency of medication for each individual patient will depend on a number of factors, including the activity of a specific derivative of fullerene, the metabolic stability and length of action, allocation rate, the patient's age, body weight, General health, sex, drug combination and the severity of the disease in the individual being treated.

For oral administration in the form of a suspension compositions prepared according to methods widely known in the field of preparation of pharmaceutical R is Ceptor, and they may contain microcrystalline cellulose or its derivatives to ensure mass, alginic acid or sodium alginate as a suspending agent, methylcellulose as an amplifier viscosity and sweetening agents and/or fragrances, known in this area. In the form of tablets such compositions may contain microcrystalline cellulose, calcium phosphate, starch, magnesium stearate and lactose and/or other excipients, binders, extenders, disintegrators, diluents and lubricants known in this field.

When applied as nasal aerosols or by inhalation, such compositions are prepared by methods well known in the field of pharmaceutical formulations, and they can be produced in the form of solutions in saline solution using benzoic acid or other suitable preservatives, promoters adsorption to enhance bioremediate and/or other solubilizing or dispersing agents known in this field.

Solutions or suspensions for injection can be formulated according to known methods using non-toxic, parenterally applicable diluents or solvents, such as mannitol, 1,3-butanediol, water, ringer's solution or isotonic sodium chloride solution or suitable dispersing or see the powers and suspendida agents, such as sterile soft stable oils, including synthetic mono - or diglycerides or fatty acids, including oleic acid.

At rectal administration in the form of candles such compositions can be prepared by mixing the drug with such a non-irritating excipients as cocoa butter, synthetic glyceride esters or polyethylene glycols, which are solid at ordinary temperatures, but sigalda and/or dissolve in the rectal cavity with the release of the medication.

The local application in the form of ointments, gels, creams, liniments, etc. such compositions can be prepared by mixing the active ingredients with acceptable ointment base.

As ointment bases can be used fat, hydrocarbon or hydrophilic bases such as vaseline, vaseline oil, paraffin wax, lanolin, polyethylene glycol, etc.

As the basis for gels can be used methyl cellulose, sodium carboxymethyl cellulose, oxypropylation, polyethylene glycol or polyethylene oxide, carbopol, polyvinylpyrrolidone, polyvinyl alcohol, etc.

The proposed invention relates to compounds, method of producing these compounds and their pharmaceutically acceptable associates with the polar reagents. The compounds do not affect l is icpoes: do not cause shear leukocyte formula and does not induce the emergence of new forms of neutrophils - neutrophilic metamyelocytes in laboratory animals (rats and rabbits). From the point of view of security (safety) this indicates the absence of these compounds toxicity that causes these changes. The inventive method allows to obtain different compositions based on fullerene-amino acids, depending on the ratio of reactants and process conditions, namely water-soluble hydrated fullerene-amino acids of General formula (II).

The method is based on the use stage of synthesis of the optimal ratio of the initial reagents, minimal quantities of organic solvent and phase transfer catalyst, followed by separation of the claimed compounds with concentrated solutions of organic and mineral acids, which leads to a quantitative obtaining fullerene-amino acid compositions of a particular composition and possible applications of the proposed method for the industrial synthesis of different efficiency and environmental friendliness.

The technical result of the proposed technical solution is to obtain a steady individual water-soluble hydrated fullerene compounds With60with aminocarbonyl acids that do not have toxic effects on the body. Developed effective way of obtaining the individual stable hydrated fullerene derivatives, possessing antiviral, antitumor and protivopsoriaticescoe activity.

The invention is illustrated by the following examples.

Example 1. Receiving, hydrate, N-fullerene-(Tris-ε-aminocaproic acid) (item IUPAC - hydrate N-fullerene-(Tris-6-aminohexanoic acid) of the formula: N-C60{NH(CH2)5COOH}3·10H2O.

To a solution of 60 g (of 0.08 mole) of fullerene C604.5 l of o-dichlorobenzene added 204 g (1.2 moles) of finely ground anhydrous potassium salt of ε-aminocaproic acid. To the resulting suspension while stirring and heating above 60°C is added during 2 hours to a mixture of o-dichlorbenzene and methyl ester of polyethylene glycol 500 in the ratio of 5:1. The reaction mixture is stirred at a temperature not exceeding 60°C for 5 hours until complete discoloration of the solution and formation of a solid precipitate. The mixture is then filtered, the filter cake was washed with several portions of ethanol and dried in a vacuum at a temperature not exceeding 60°C. the Selected mixture of potassium salts of fullerene-aminohexanoic acid and aminohexanoic acid are dissolved in 100 l of distilled water. To the solution slowly with stirring, add 0.1 N hydrochloric acid to a pH of 5.1. The mixture defend to complete planting of the product, Then the aqueous layer was decanted. Sediment predstavlyayushie is a fine suspension of solid product in water, centrifuged and washed with water to pH 6. The precipitate is dried at a temperature not above 60°C in a vacuum drying Cabinet.

The yield is quantitative and is 115,

The connection is a dark-brown solid, soluble in water, soluble in mixtures of CH3CN:H2O - 1:10 and DMF:H2O - 1:100.

According to thermogravimetric analysis of the compound obtained contains 10 moles of H2O. At a temperature of 350°C results in intense destruction complex. The residue after decomposition contains fullerene and its oxidation product.

The IR spectrum of the product (I) contains absorption bands, characteristic of N-substituted amino acids: a group-COOH - 1704 cm-1, 1658 cm-1N-H-stretching vibrations 3400 cm-1N-N-deformation - 1552 cm-1the absorption bands of C60-NH-R - 1104 cm-1, 930 cm-1, 830 cm-1.

Electronic absorption spectrum does not contain absorption bands of free fullerene.

Elemental analysis of the compound shows the following ratio of components:%=72,75; % H=4,70; % N=2,32; calculated for gross formula C78H39O6N310H2O: % C=72,38, % N=4,3, % N=3,24.

The number of carboxyl groups in the product was determined by reaction with metal salts and amines. By the reaction with silver nitrate is quantitatively highlighted the complex composition of C60(H)3 {NH(CH2)5COOAg}310H2O. (Found: % Ag=to 20.88, % C=57,80, % N=Of 2.51, % N=3,32; calculated: C78H36O6N3Ag3(10H2O) - % Ag=20,00, % C=57,88, % N=2,60, % H=3.46 In).

By reaction with trisamino obtained water-soluble compound C60(H)3{NH(CH2)nCOO-NH3+C(CH2OH)3}3(found % C=64,88, % N=4,56, % N=5,08 calculated for C90H72O15N610H2O: % C=65,2, % N=4,34, % N=5,10).

Example 2. Receiving, hydrate, N-fullerene-(Tris-ω-aminoalkanoic acid) (item IUPAC - hydrate N-fullerene-(Tris-7-aminoheptanoic acid) of the formula: N-C60(H)3{NH(CH2)6COOH}3·8H2O.

To a solution of 72 g (0.1 mole) of fullerene C604 l o-dichlorobenzene added 182 g (1.2 moles) of finely ground anhydrous potassium salt of ω-aminoalkanoic acid. To the resulting suspension with stirring and heated above 80°C are added in the course of 3 hours a mixture of o-dichlorbenzene and methyl ester of polyethylene glycol 500 in the ratio of 5:1. The reaction mixture is stirred at a temperature not exceeding 80°C for 8 hours until complete discoloration of the solution and formation of a solid precipitate. The mixture is then filtered, the filter cake was washed with several portions of ethanol and dried in a vacuum at a temperature not exceeding 60°C. the Selected mixture of potassium salts of ullern-aminoanisole and aminoalkanoic acid are dissolved in 120 l of distilled water. To the solution slowly with stirring, add 0.1 N hydrochloric acid to a pH of 5.1. The mixture defend to complete planting of the product, then the aqueous layer was decanted. The precipitate, which is a fine suspension of solid product in water, centrifuged and washed with water to pH 6. The precipitate is dried at a temperature not above 60°C in a vacuum drying Cabinet.

The yield is quantitative and is 130,

The connection is a dark-brown solid, soluble in water, soluble in mixtures of CH3CN:H2O - 1:10 and DMF:H2O - 1:100.

According to thermogravimetric analysis of the obtained compound contains 8 moles of H2O. At a temperature of 450°C results in intense destruction complex. The residue after decomposition contains fullerene and its oxidation product.

The IR spectrum of the product contains absorption bands, characteristic of N-substituted amino acids: a group-COOH - 1707 cm-11650 cm-1N-H-shaft. fluctuations - 3400 cm-1N-H-deformation - 1552 cm-1the absorption bands With60-NH-R - 1104 cm-1, 930 cm-1, 830 cm-1.

Electronic absorption spectrum contains absorption band at 260 nm.

Elemental analysis of the product shows the following ratios of elements: % C=to 73.55; % H=4,60; % N=3,18; calculated values for gross formula C81H45O6N3 2O) - % C=74,82, % N=4,69, % N=3,23.

By the reaction with silver nitrate selected silver salt fullerene-amino acids, quantitatively demonstrating the presence of three amino acid fragments in the composition of the obtained product.

Example 3. Receiving, hydrate, N-fullerene-(Tris-8-aminooctanoic acid) of the formula: N-C60(H)3{NH(CH2)7COOH}3·10H2O.

Carried out analogously to example 1 but instead of finely ground anhydrous potassium salt of ε-aminocaproic acid (ω-aminoalkanoic acid) use potassium salt aminooctanoic acid. Analysis of the obtained compound proves represent the complex.

Was studied the antiviral activity of compounds against HIV, HSV, influenza virus, and anti-tumor activity. The connection has a high antitumor and antiviral activity against all these viruses. Below are the best examples of embodiment of the invention. In the examples below, the compound obtained by the method described in example 1, is referred to in the text preparation No. 1 (fullerene-Tris-aminocaproic acid hydrate).

Example 4. Studying the activity of fullerene-Tris-aminocaproic acid against human immunodeficiency virus.

The studies were conducted in the Institute of Virology. Dijanoveckog RAMS, Moscow. In the task, and the research was to study the activity of the drug against human immunodeficiency virus.

To the cells was added to the study drug and were infected with virus at a dose of 0.01 TCID50/cell. Incubated cell culture at 37°C in an atmosphere with 5% CO2and 98%

humidity 4-5 days. Analysis was performed by staining cells with dye and light microscopy: study of the cytopathic effect of the virus (JRC) and virusinduced entityarray (syncytium - a conglomerate of several cells from the total cell membrane, formed by the merger of their membranes).

The degree of cytodestructive was evaluated under a microscope by the common chetyrehrazovoe the system by the signs + or - respectively the number of dead cells in each of the four holes corresponding to one of the investigated parameter.

++++ - 100%thcell death in four holes used in the experience of one breeding

+++ - 75%thcell death in each of the four holes,

++ - 50%thcell death in each of the four holes,

+ - 25%thcell death in each of the four holes,

+- - beginning degeneration

- no cytodestructive.

The results of the study are presented in tables 1-2.

The data obtained (table 1, 2) showed that the drug No. 1 has antiviral activity against human immunodeficiency virus type 1 in a concentration of 1-10 μg/ml EC50(50%efficiency is Naya concentration) of the proposed drug 5,0 µg/ml.

Example 5. Studying the activity of fullerene-Tris-aminocaproic acid against influenza virus.

The studies were conducted in the Institute of Virology. Dijanoveckog RAMS, Moscow. The objective of the research was to study the antiviral activity of the drug in the culture of MDCK cells against influenza virus A/IIV-Moscow/01/2009 (H1N1)sw1.

The drug was dissolved in dimethyl sulfoxide (DMSO) (5 mg substance + 0.5 ml DMSO) followed by the addition of 4.5 ml of medium for cell cultures MEM, receiving thus the flow at a concentration of 1.0 mg/ml In subsequent conducted breeding wastewater environment MEM to working concentrations of 6.5 μg/ml and 12.5 - 25,0 - 50,0 - 100 µg/ml

Determination of the antiviral activity of the substances was performed to reduce the reproduction of influenza virus in cell culture MDCK detected by ELISA.

With this purpose, cells MDCK were grown in 96-well tablets up to a full monolayer was washed from the growth medium and added substances in double concentration in 100 μl of MEM medium. Infection with the virus in the working dose of 100-1000 TCID50conducted in two modes: 2 hours after introduction of substances and simultaneously. The plates were incubated in an incubator with CO2for 24 hours at 37°C. After incubation the medium was removed and cells were fixed with 80% acetone in PBS for 15 minutes, well dried and staged ELISA, by conducting successively the adsorption of the special is practical reagents monoclonal antibody, conjugate and substrate (orthophenylphenol). The reaction was accounted by optical density at 492 nm on a spectrophotometer firm "Biocom". Each dilution of virus was investigated in 3 repetitions, for which the calculated average value of the optical density (OD). The percentage of inhibition was determined as the ratio between the difference OP OP experience and the cell control, divided by the difference OP virus control and es cell control, multiplied by 100%. On the basis of the data obtained, the value of the minimum concentration of a substance that causes 50.0% inhibition of viral reproduction (MIC50).

Assessment of the suppression of the reproduction of the virus of influenza A(H1N1) was performed in 3 experiments at different multiplicity of infection. The results are presented in table 3 (protocols 3 experiments) and table 4 (average values of the obtained results of 3 experiments).

As can be seen from table 4, the greatest activity to reduce the reproduction of influenza virus in cell culture MDCK showed a series of preparation 1. Clearly the degree of reproduction and the concentration of the drug: with increasing concentration of the reduced reproduction of the virus. In addition, significant differences in performance in different modes of infection (2 hours after making the drug or simultaneously) is not marked. Indicators of minimum ingibirujut is th a reproduction of the virus in 2 times concentration (MIC 50)amounted to: in the mode of making the drug for 2 hours to 9.5 μg/ml, and in simultaneously making - 12.5 µg/ml. the Calculation is performed from the graphical construction of received data.

Thus, the results obtained studying the activity of different batches of product No. 1 against influenza virus A/IIV-Moscow/01/2009 (H1N1)sw1 revealed high activity of suppressing its reproduction in cell culture MDCK series 1 with an average activity of series 2. While the modes of making preparations for 2 hours prior to infection or simultaneously with infection had no effect on their activity in cell culture MDCK.

Example 6. Study of antiviral activity of fullerene-Tris-aminocaproic acid on the model of influenza pneumonia in mice.

The study was performed in the center of the chemistry of drugs (CHLS - UNIFI), Moscow.

In this work, we used the drug No. 1 in the form of a dark brown powder. For oral administration prepared the necessary dosage, dissolving the sample in a 1% solution of starch, boiled water. For intraperitoneal and intramuscular administration sample preparation step 1 was dissolved in a 1.5% solution of dimethyl sulfoxide.

The work was used influenza virus A/Aichi/2/69 (H3N2), adapted to mice. This virus is widely used to determine the effectiveness of antiviral drugs on the model of influenza pneumonia we who she is and was obtained from the Museum of viral strains and cell cultures Institute of Virology RAMS. For the preparation of infective material, the mice were infected intranasally allantoin virus, after developing signs of disease were scored and in sterile conditions received the homogenate of lung tissue. Next, the homogenate was used to infect 10-day-old chick embryos, from which received allantoine virus and after titration his mice was used to infect the animals.

Outbred mice (females) weighing 12-14 g were obtained from the kennel "Andreevka" (Moscow region) and were kept on a standard diet in regulated vivarium conditions.

Weighted mouse (female non-linear, the average weight of 12-14 g) were infected intranasally under light ether anesthesia influenza virus A/Aichi/2/69 (H3N2) (10 LD50100 µl). In the preliminary experience carried out a determination of LD50by titration allantoinase virus such as mice, which are then used in the main experience. Was used following treatment with investigational drug: 24 hours prior to infection, 1 hour before infection, after 24 hours, and then 1 time per day after 24 hours for 5 days. For oral administration used disposable insulin syringe with a special needle (lavage), each dose was administered in a volume of 100 μl. For intraperitoneal and intramuscular treatment also each dose was administered to voyame 100 μl. The virus control group consisted of 10 mice infected with the virus, but not treated with drugs. Also in the experiment had two groups of 10 uninfected mice, which were injected intraperitoneal and intramuscular injection of 100 μl of 1.5% DMSO, which was used as a solvent preparations. In other groups also initially contained 10 animals. For treated and control animals was performed daily observation, in the first 5 days after infection mice were weighed every day, after day. Chemotherapeutic drug activity No. 1 on the model of influenza pneumonia mice was assessed by three criteria: a measure of protection against lethal viral infection, increasing life expectancy and reducing weight loss in the groups of animals treated with the drug compared with the control group.

Drug treatment No. 1 was effective, reducing the mortality of mice against influenza pneumonia and loss their weight and increasing life expectancy compared to the virus control. The effectiveness of this treatment depended on the dose and method of treatment. The effectiveness of oral treatment fullerene-Tris-aminocaproic acid hydrate was increased with increasing dose. Oral drug treatment No. 1 was effective, increasing life expectancy B1,6-1,7 times. The most effective in all three parameters, a measure of protection from mortality, life expectancy and weight loss) was the treatment of fullerene-Tris-aminocaproic acid hydrate intramuscularly in doses of 100 and 200 mg/kg/day prevented the loss of 70-80% of infected animals and the loss of their weight, and increased their life expectancy by almost 2 times.

Intraperitoneal treatment of fullerene-Tris-aminocaproic acid was effective only at doses of 50 and 100 mg/kg/day. The death of animals, a significant reduction in life expectancy and weight of mice after intraperitoneal treatment of their drug No. 1 in a dose of 200 mg/kg/day suggest that this dose in this method of introduction is toxic to infected mice. The results are presented in tables 5-6.

Example No. 7. Study of the protective activity of fullerene-Tris-aminocaproic acid in experimental lethal influenza infection in white mice, caused by viruses of different origin.

The study was performed in the research Institute of influenza, St. Petersburg.

In this work, we used the drug No. 1 in the form of black powder. Sample preparation were dissolved in the medium for cell cultures Needle MEM (Biolot, St. Petersburg, cat. No. 1.3.3). From the resulting solution were prepared with the ' series of dilutions in MEM medium to determine the antiviral activity of the samples in experiments on animals.

As reference drugs used Rimantadine (1-(1-substituted)-amino-ethyl hydrochloride, Aldrich Chem. Co., Milw., WI, cat. No. 39.059-3) and Tamiflu (Ethyl(3R,4R,5S)-4-acetamido-5-amino-3-(1-ethylpropoxy)-1-cyclohexen-1-carboxylate phosphate, Hoffmann LaRoche, Switzerland).

The viruses. In the work were used-adapted mice influenza viruses following strains:

- A/Swine/1976/31 (H1N1) - swine origin;

- A/Puerto Rico/8/34 (H1N1) human origin (rimantadine-resistant);

- A/Vladivostok/2/09 (H1N1) human origin (Tamiflu-resistant). Viruses were passively in allantoine cavity 10-12 day old chick embryos for 48 hours at 36°C. the Strain And/Vladivostok/2/09 (H1N1) pre-adapted to mice by three pairs of alternating passages in animals and in chicken embryos.

For infection of animals was used vaccinated allatoona fluid of chicken embryos. It was prepared by serial 10-fold dilutions in saline solution, after which the contagious virus infects the material was determined in a separate experiment by titration on mortality in animals. The titer of the virus was calculated by the method of reed and turns into a hissing drone (Am. J. Hyg., 1938, 27: 493-497).

Outbred mice (females) weighing 14-16 g were obtained from the kennel "Rapolano" (Leningrad region) and were kept on a standard diet in DHL is montirovannyh vivarium conditions of the research Institute of influenza, Russian Academy of medical Sciences. Selection of animals in the group experiment was carried out by random sampling. Before testing, the animals were under observation for 2 weeks.

The study drugs were administered to the animals intraperitoneally in a volume of 0.2 ml in the following doses: Drug No. 1 - 300, 100 and 30 mg/kg, Rimantadine - 50 mg/kg, Tamiflu 20 mg/kg of animal weight. The drugs were injected by medical scheme: for 24 hours and 1 hour before infection and after 24, 48 and 72 hours after infection. The placebo control group animals were injected with physiological phosphate buffer. As a negative control was used intact animals, which were kept in the same conditions as the experimental group.

Viruses were injected animals intranasally under light ether anesthesia in a dose of 1 and 10 LD50. In each group the observations were taken on 25 mice. On day 3 after infection, 10 animals from each group were killed, dissected and isolated lungs. Of these 10 light 5 used for isolation of the virus (frozen and kept at -20°C prior to the formulation of the relevant experiments), while the remaining 5 were fixed with 10% formalin and used for histological analysis (see below).

Monitoring of the remaining animals was carried out within 14 days, i.e. the period during which in experimental influenza observed mortality of animals. Daily recorded mortality belly who's in control and experimental groups. On the basis of the mortality in each group was calculated percentage mortality (M, the ratio of the number of the fallen for 14 days the animals to the total number of infected animals in the group), the index of protection (IP, the ratio of the difference in the percent mortality in the control and experimental groups for the percentage of mortality in the control group) and the average life expectancy of animals (DL) is based on 14 days of observation.

Animals survived to 15 days after infection, were opened and visually evaluated the vastness of foci-borne pneumonia in the lungs. The size of the lesions was expressed in percent of the total surface of the lungs.

Clinical signs of disease were typical of influenza infection and include difficulty breathing, ataxia, tremor, and reduced food and water consumption and, consequently, of animal weight.

Data on the dynamics of mortality of animals in the control and experimental groups are summarized in tables 7-9.

As can be seen from the presented results, the influenza virus caused a lethal infection in white mice, accompanied by the death of animals ranging from 3-4 days after infection, depending on the dose of virus. In this respect the lives of animals, has been associated with the used dose of virus reverse dependence. Rimantadine used in the experiment as the reference drug, was assisted in this in the eccii very modest protective effect, that was manifested by a slight decline in mortality in the experimental groups compared with control (index of protection 13-29%) and a slight increase in life expectancy (1.1-1.6 days depending on the dose of virus). The resulting data, therefore, are consistent with previously obtained results of experiments in vitro and in vivo, indicating the insensitivity of the used strain of the virus to rimantadine. Some protective effect in this case it is possible to explain the toxic effect of the drug.

At the same time, the comparator drug Tamiflu exhibited a pronounced protective effect as reducing mortality in the groups of mice treated (approximately 70% compared with control)and increasing the average life of animals (2-6 days). Thus, used the virus was resistant to rimantadine, however, sensitive to Tamiflu.

The analysis of the obtained data it was found that the investigated sample preparation for its protective properties were close to the reference preparation Tamiflu (table 7).

The obtained results were confirmed by using the model of influenza pneumonia caused two other influenza virus strains. The data from these experiments are summarized in tables 8-9.

As can be seen from the above data, the activity of chemotherapy in relation used VI the whiskers were significantly different. So, in relation to the influenza strain A/Vladivostok/2/09 causal drug Tamiflu has been inactive. Thus, the previously obtained data about the sustainability of this isolate to Tamiflu have been confirmed in animal experiments. At the same time the activity of the investigational drug against this strain was very high, which certainly should be considered as an advantage of the drug.

The rate of activity of the investigational product (index of protection - life) amounted to - 21-72% and 0.8 and 4.4 days depending on the applied strain infecting virus dose and dosage.

To study the influence of the drug No. 1 on the replicative activity of influenza virus in the lung tissue of infected animals at 3 days after infection of the lungs of animals were prepared homogenates, which then determined the infectious titer of the virus in cell culture. Level data replication model of influenza viruses in animals is given in table 10.

As can be concluded from the presented results, all three used the virus was able to replicate in the lung tissue of mice, reaching to the day 3 titles 3,4-6,4 log10EID50/20 mg depending on the applied strain and the infective dose of the virus. The use of chemotherapy - study drug and Comparators is limited to the Ivalo multiplication of the virus in different degrees. So, rimantadine insignificant (by 2-3 orders of magnitude) reduced infectious activity of sensitive virus A/Swine/1976/31 and/Vladivostok/2/09, however, did not show a reliable inhibitory activity against rimantadine-resistant strain A/Puerto Rico/8/34. Tamiflu was active against virus A/Swine/1976/31 and A/Puerto Rico/8/34. At the same time when testing it on a sustainable model of strain And/Vladivostok/2/09 was found some reduction in infectious titer of the virus, however, differences from the control were false.

The preparation of the study showed significant inhibitory activity against all the studied viruses. The level is not exceeded, however, was comparable to the activity of the Comparators of Rimantadine and Tamiflu. When using viruses that are resistant to chemotherapeutic drugs, the activity of the study drug was significantly higher than the activity of Tamiflu against oseltamivir-resistant strain And/Vladivostok/2/09 and the activity of rimantadine versus rimantadine-resistant strain A/PR/8/34.

When studying the characteristics of morphogenesis experimental influenza infection in treatment-and-prophylactic medication was No. 1 at the dose of 300 mg/kg was noted that the morphogenesis of infectious process in the lungs of animals treated with the drug, largely differed from morphological changes in the lungs control the stomach who's. The main difference on the 3rd day after infection related to the nature of the inflammatory exudate, namely, that with the same intensity it practically not observed cells in the stage of decay characteristic of the acute stage of influenza pneumonia. The cellular component of the exudate was represented exclusively by intact neutrophils, lymphocytes and macrophages. In addition, serous and hemorrhagic components of the exudate were also expressed weaker. Cells of the bronchial epithelium looked more intact than in control animals. Themselves inflammation took less compared with the control area.

The same trends were observed at the stage-borne pneumonia. Lesions of the lungs were severely limited in size, the morphological examination revealed mild metaplasia of the epithelium and infiltration interstice intact neutrophils and round-cell elements. It should be noted that the effect of the drug was observed in infected animals in any of the three studied viruses regardless of their sensitivity or resistance to drugs comparison.

An additional criterion protective of drug action No. 1 served as the evaluation of lesions of chronic pulmonary lesions in animals. The results of this test are shown in table.

As can be seen from the bring is the R results all three viruses induced the formation in the lungs of persistent foci of chronic lesions detected visually in surviving animals on day 15 after infection. Comparators - rimantadine and Tamiflu - significantly reduced the length of foci-borne pneumonia caused by susceptible to viruses, and were inactive in the case of resistant strains. At the same time, the product No. 1 was significantly reduced this figure is used regardless of the virus.

Thus, it is shown that at the studied concentrations (300-30 mg/kg) product No. 1 shows dose-dependent protective activity on used models. This activity was shown in the following figures:

- 6-200-fold reduction in infectious titers of virus in the lung tissue of infected animals;

- prolong the life of infected animals (0.1-4.4 days depending on the applied strain, virus dose, party synthesis and dosage);

- reduction of specific mortality in the groups experience on 7-72% depending on the applied strain, virus dose, party synthesis and dosage;

- reduction of 2-4 times the average length of foci of chronic borne pneumonia.

The main result of the protective activity of the drug is No. 1 at some doses were comparable with the activity of the drug cf is Vania - of rimantadine.

The data obtained indicate the presence of the drug No. 1 high influenza activity, including against strains of swine origin, and also against viruses that are resistant to applied in the clinic influenza drugs Rimantadine and Tamiflu.

Example 8. The study of antitumor activity of the drug fullerene-Tris-aminocaproic acid in models of solid and ascitic form of Ehrlich carcinoma in white mice.

The objectives of this study were:

- study of the effect of drug # 1 on the growth of ascitic tumors intraperitoneal injection of cancer cells;

- the study of drug action No. 1 on the growth of solid tumors, the study of the effects of drugs on the morphology and morphometric indices of the solid form of Ehrlich carcinoma;

- the study of drug action No. 1 on the apoptotic activity of cells of ascitic form of Ehrlich carcinoma.

In this work, we used an aqueous solution of the drug No. 1 in two doses at concentrations of 30 and 10 mg/kg, the Animals were injected subcutaneously with 0.2 ml of each concentration for 24 hours before inoculation and then daily for the entire period of the experiment. The final drug concentration was 300 and 100 mg/kg of body weight.

As the comparison drug used Cisplatin - protivoopujolevy is a new drug, practices used in cancer therapy person. Cisplatin was administered once on day 2 after inoculation of the tumor, because of its high toxicity. The final concentration of Cisplatin was 5 mg/kg of body weight.

The experiments were carried out on outbred mice average weight of 20±3 g (animal farm, Rappolo, Len. region).

Cells of Ehrlich carcinoma were obtained from the Museum of cell lines Oncology research Institute and cultivated in the abdominal cavity of mice. For this, 0.2 ml of cell suspension was administered to the animals intraperitoneally. 7-10 days after inoculation, animals were killed, ascitic fluid was collected via puncture of the peritoneum, diluted saline solution 10 times and placed on ice.

In order to simulate the solid form of Ehrlich carcinoma, the animals were injected subcutaneously in the region of the right femur 0.2 ml of cell suspension for 40 minutes after collecting ascitic fluid placed on ice. In the course of experience within 28 days, twice a week starting 8 days after inoculation, a measurement was performed tumor nodules using a micrometer. The tumor size was calculated by multiplying half the length of the bundle on a square of width expressed in mm3. Also recorded a mortality of animals in the control and experimental groups. On the 29th day after inoculation, animals were slaughtered.

To study the effects of drugs nascita form tumors, the animals were injected intraperitoneally with 0.2 ml of cell suspension for a period of not more than 40 minutes after collection of the initial ascitic fluid. During the experiment was carried out weight control mice as an indicator of the accumulation of ascitic fluid in the abdominal cavity. Observation of the animals was carried out for 16 days. On the 17th day after inoculation, animals were slaughtered.

Data on the dynamics of tumor growth and mortality of animals from the ascitic form of carcinoma in the control and experimental groups are presented in table 12.

As can be seen from the above results, the inoculation of tumor cells into the peritoneal cavity of animals caused a rapid accumulation in the cavity ascitic fluid. The use of therapeutic drugs had a pronounced therapeutic effect and resulted in inhibition of the accumulation of ascites.

Treatment of animals with the proposed drug and the comparator drug is Cisplatin, which led to significant inhibition of the dynamics of weight gain of the animals. In the later stages of the development process of these differences reached statistical significant values.

The effect of the drug on the process of apoptosis in cells of medium size and the granularity of the ascitic form of Ehrlich carcinoma in white mice are shown in tables 13-14.

It follows from the presented results, only a small fraction of the cells in both tumor subpopulations were being reversible or irreversible apoptosis in control animals. The use of cisplatin led to a sharp increase in the proportion of cells in R is na stage of apoptosis among immune cells (table 13) and late apoptosis and necrosis among tumor cells (table 14).

Preparation No. 1 acted on the level of Cisplatin: like a drug comparison, he stimulated early apoptosis in immune cells and late apoptosis in a subpopulation of tumor cells. Living (AnV-7AAD-) tumor cells under the influence of the drug remained. The level of induction of necrosis in tumor cells it is even superior to Cisplatin (30,2% versus 26.6% for Cisplatin). The mechanism of anticancer drug action No. 1 and the activity of Cisplatin, was the induction of apoptotic processes in tumor and immune cells, components of ascites. The process of apoptosis was selectively and significantly faster in tumor cells than in neutrophils and lymphocytes, found in ascitic fluid.

The results cytofluorometric analysis allow to speak about the selectivity of drug action comparison - Cisplatin on tumor cells compared with normal cells, also present in the composition of the ascitic fluid. At the same time after administration of the drug to normal cells, which constitute a fraction of the average size and granularity - neutrophils, macrophages, lymphocytes, and others - were in early apoptosis, and tumor cells - late (irreversible) apoptosis or necrosis.

Data on the dynamics of the development of solid tumors under Asteem study drug and drug Cisplatin in comparison with control group of animals is presented in table 15.

As can be seen from the presented results, all used drugs in varying degrees retarded tumor growth throughout the experience. In General, the most pronounced antitumor effect had Cisplatin. Significant inhibition of tumor growth was observed when applied up to 17 days after inoculation.

At the same time, both series of the drug also showed a dose-dependent antitumorigenic effect. A significant decrease of tumor size and inhibition of growth was observed up to 8 days of the experiment. In the future, the drugs also inhibited tumor growth in all periods of the study, although the differences with the control and did not reach statistical significance. In General it should be noted drug No. 1 in a dose of 100 mg/kg of body weight as the most similar in effectiveness to this drug called cisplatin in almost all periods of the experiment.

The obtained results do not allow us to consider the drug No. 1 as the leading drug for targeted therapy of cancer. However, on the basis of these data we can talk about it as promising for the further development of the tool integrated treatment for co-use with other drugs, particularly when ascitic forms of tumors.

Pharmaceutical form of the proposed drug can be used orally, parenter the flax (including subcutaneous injections, intravenous, intramuscular, vnutrigodovye injection or infusion), by inhalation spray or rectally for the treatment or prophylaxis of viral infections such as HIV, herpes, influenza of different origin, as well as anticancer drugs for comprehensive treatment.

Compounds mixed with conventional pharmaceutical carriers and excipients and used in the form of tablets, capsules, suppositories, ointments, emulsions, solutions, sprays. It should be noted that for the preparation of solutions, sprays, as well as soft dosage forms (ointments, suppositories) connection pre-diluted in a mixture of DMSO with water.

The treatment of infectious diseases by influencing pharmaceutically acceptable doses of the compounds according to formula II is carried out simultaneously for several viruses (in the case of mixed infections) and affects different stages of viral replication. It is shown that the treatment is accompanied by a reduction of stress effect on the introduction of the drug, increased antioxidant protection of the organism against infection, excretion of toxins. Intoxication characteristic for the flow of a number of viral infections and determines the severity of the disease.

The possible combinations of the compounds of the formula II with other antiviral agents, immunomodulators, anti-infective agents or VA is the care in various combinations with any of the pharmaceutical compositions intended for treatment.

Example 9. The study of chronic toxicity of fullerene-(Tris-aminocaproic acid) hydrate in rats with intramuscular injection within 30 days.

The experiment was conducted in fgun "research center for toxicology and hygienic regulation of biopreparations" (fsri SRC TBC coatings of the FMBA of Russia), Serpukhov.

The aim of the study was experimental evaluation of the level and nature of the possible damaging effect of fullerene-(Tris-aminocaproic acid) hydrate on the body of rats with intramuscular injection within 30 days.

Experiments were performed on Wistar rats, purchased in the nursery GU NCBT RAMS (branch "Column"). The animals corresponded to the sanitary rules approved by the USSR Ministry of health 06.07.73,, device, equipment maintenance experimental biological clinics (vivarium). Fed belly what's natural and piketirovany feed in accordance with the standards approved by order of the USSR Ministry of health No. 755 from 12.08.77, the Animals were quarantined and acclimatized in vivarium conditions for 5 days.

Experimental groups of animals were formed by random sampling with the body mass index as a leading indicator.

The investigated substance was administered to rats intramuscularly daily for 30 days at doses of 3, 9 or 20 mg/kg in the form of solutions of different concentrations in a 20% solution of dimethyl sulfoxide (DMSO). Animals of the control groups were administered a 20% solution of DMSO. Working solutions of the substance and DMSO were prepared daily immediately before use. Enter the volume of the doses were adjusted to suit the individual animal body weight after each weighing. Each dose was tested on 20 animals (10 females and 10 males).

For the evaluation of toxic effects of fullerene-(Tris-aminocaproic acid) hydrate 24 hours after completion of the course introduction substance half of the animals from each group were taken out of the experiment to conduct hematological, biochemical and pathological studies. The second half of the experimental animals were taken out of the experiment after the cancellation period the introduction of the substance and conducted similar studies.

In the period of introduction of the substance and within 14 days after its abolition was daily assessed the General condition and clinical symptoms of intoxica the AI animals. The General condition of the animals was assessed by their physical activity, consumption of food and water, wool and visible mucous membranes, body weight.

Hematological analysis was performed using semi-automatic dual-channel conductivity meter cells Hema-screen 13 (Hospitex Diagnostics, Italy), as well as using light microscopy.

Biochemical indices of blood serum was determined using a semi-automated analyzer, "Stat Fax 3300".

Biochemical parameters of urine was determined using a semi-automated analyzer Urisys 1100".

The morphological state of the internal organs of the animals was determined visually at postmortem autopsy and microscopic study of histological specimens (paraffin sections of 4-5 μm, stained with hematoxylin and eosin).

Statistical analysis of the received results of research conducted by methods of variation statistics by student's criterion.

1. The results of the research.

1.1. The results of clinical observation.

Rats daily for months intramuscularly injected with a fullerene-(Tris-aminocaproic acid) hydrate in doses of 3, 9 or 20 mg/kg At all doses, the clinical symptoms were absent, the General condition of the animal experimental and control groups did not differ. The weight gain of rats in the course in which his experience in experimental groups was not significantly different from the control (table 16).

1.2. The results of biochemical analysis of blood serum

After the introduction of fullerene-(Tris-aminocaproic acid) hydrate in male rats shown significant reduction in the level of urea at the maximal tested dose (table 17). Shows changes in the concentration of cholesterol in the minimum and average dose is not associated with the action of the investigated substances and do not go beyond the limits of physiological norm. In female rats shows a slight but significant increase in the activity of alanine aminotransferase at the maximal tested dose. The revealed changes of the glucose level at the minimum dose, the concentration of total protein and cholesterol on the average dose of a substance is not have dose-response relationships and do not go beyond the limits of physiological norm.

A significant decrease in creatinine concentration at the maximum tested dose shown at the end of the cancellation period fullerene-(Tris-aminocaproic acid) hydrate in male and female rats. At this same dose shows the change in aspartate aminotransferase activity and in male and female rats, however, in the case of males is reduced activity, while females increased activity (table).

Evaluating the results of the analysis of biochemical parameters of blood serum of rats during the d introduction substance and after the cancellation period introduction it should be noted that the values of the above changes of the serum values of female and male rats do not extend beyond the physiological range for the species.

1.3. The results of hematological analysis of blood

After the introduction of fullerene-(Tris-aminocaproic acid) hydrate revealed no changes in hematological parameters associated with the test substance. So in female rats installed slight decrease in the average volume of an erythrocyte mid-dose and a slight increase in the proportion of reticulocytes on the minimum dose (table 19). These changes do not go beyond the limits of physiological norm and not have dose-response relationships.

The study of hematological parameters of rats at the end of the cancellation period fullerene-(Tris-aminocaproic acid) hydrate revealed no significant differences between the control and experimental animals (table).

1.4. The results of urine analysis

At the end of the period of the introduction of fullerene-(Tris-aminocaproic acid) hydrate was found to be increasing urine pH in the group of males at the dose of 9 mg/kg (table 21). The level change indicator does not go beyond the physiological norm, dose dependency does not exist.

After the cancellation period of the introduction of fullerene-(Tris-aminocaproic acid) hydrate in the group of females at a dose of 20.0 mg/kg marked decrease in the relative density of urine compared with the control group animals. In groups of males in the same period, there are statistically significant differences in comparison with the control group: at the dose of 3.0 mg/kg - increase in pH, at a dose of 9.0 mg/kg - increase in relative density of urine. Both indicators do not go beyond the limits of physiological norm and not have dose-response relationships (table 22).

1.5. The results of pathological studies

At postmortem autopsy carried out after the end of the period intramuscular fullerene-(Tris-aminocaproic acid) hydrate, when the external examination has not been established differences between rats in the experimental and control groups: the coat was smooth, shiny, leather, elastic, lively, subcutaneous tissue moderately pronounced, visible mucous membranes pale, pure, without ulceration and foreign overlays, pathological discharges from natural orifices of the body was missing. At necropsy of the thoracic and abdominal cavities were observed anatomically correct positioning of the internal organs. Macroscopically visible signs of pathology within the unit bodies are not installed. When the dissection of skeletal muscle force group (injection) in animals treated with fullerene-(Tris-aminocaproic acid) hydrate at all tested doses, noted brownish color muscle tissue, fascia, and fat layers. Animals of the control group of such color these tissues was not found.

In the analysis of the mass coefficients of the internal organs of animals after a period of intramuscular injection of fullerene-(Tris-aminocaproic acid) hydrate was not established differences between rats in the experimental and control groups (table).

Microscopic examination was performed a comparative assessment of the histopathological picture of the organs and tissues of animals treated with fullerene-(Tris-aminocaproic acid) hydrate at the maximum dose, and rats of the control group. In the analysis in both groups were identified individual pathological changes were found mainly in the lungs, liver and kidneys (table). The degree of detected changes slightly varied within groups, but mainly corresponded to weak or moderate. Given the absence of any alternative or proliferative response in the areas of detected changes in the organs, as well as a large number of cases of acute hyperemia of blood vessels in them, the more likely the reason for their occurrence is the individual response of the animals to General anesthesia and inhalation of carbon dioxide euthanasia. Based on approximately the same frequency of occurrence installed pathological changes in experimental and control groups, we can make a conclusion about the absence of induction of the investigated substance. In this regard, these changes were considered as background.

In the introduction fullerene-(Tris-aminocaproic acid) hydrate - skeletal muscle were detected changes as inflammatory nature (small foci of hemorrhage, lesions compressed and swollen muscle fibers and infiltration of lymphoid cells in the space between the individual muscle fibers and fiber bundles)and regenerative (lots of loose or dense connective tissue with newly formed blood vessels). These changes were inherent and control group rats. This pattern is common to multiple repetitive injury as a result of intramuscular injections in this study - regardless of introduced substances.

At postmortem autopsy after the cancellation period of the introduction of fullerene-(Tris-aminocaproic acid) hydrate at external examination did not reveal differences between the rats in the experimental and control groups. Brown staining of the tissues at the site of a previous injection fullerene-(Tris-aminocaproic acid) Hydra is not detected. In the statistical analysis of the mass coefficients of the bodies was not established significant differences between experimental and control animals (table).

Microscopic examination of histological preparations bodies detected changes in the nature and severity mainly corresponded described after the introduction fullerene-(Tris-aminocaproic acid) hydrate, and also, about the same degree, were inherent in an animal experimental and control groups (table). Therefore, in this case, the conclusion was made about the lack of induction of the revealed changes of the investigated substance.

In the injection study and control substances - skeletal muscle - installed similar for experimental and control animals changes: few thin stretches of fully formed dense fibrous connective tissue, located either along the muscle fibers, or at an acute angle thereto. This morphological pattern indicates the absence of differences in the speed and nature of the healing process in injection solvent and fullerene-(Tris-aminocaproic acid) hydrate.

Thus, the results of postmortem studies of morphological traits associated with who is esteem or cancellation of fullerene-(Tris-aminocaproic acid) hydrate is not installed.

Conclusion. Throughout the period of the introduction of fullerene-(Tris-aminocaproic acid) hydrate in rats and the cancellation period for introducing the substance was not observed signs of changes in the clinical condition of the animals. The introduction of fullerene-(Tris-aminocaproic acid) hydrate did not affect the behavior, the condition of the coat, visible mucous membranes and weight gain in experimental animals.

Long-term within 1 month intramuscular administration of fullerene-(Tris-aminocaproic acid) hydrate in rats had no effect on the peripheral blood. Cancel the introduction of a test substance did not cause changes in blood parameters.

After the introduction of fullerene-(Tris-aminocaproic acid) hydrate in male rats shown significant within physiological norms decrease in the level of urea at the maximal tested dose (20 mg/kg), and in female rats shows a slight but significant increase in the activity of alanine aminotransferase. The decrease in the concentration of creatinine in the maximal tested dose shown at the end of the cancellation period of the investigated substance in male and female rats.

When analyzing the results of the examination of the urine of rats after the cancellation period of the introduction of fullerene-(Tris-aminocaproic acid) Hydra is in the group of females at the dose of 20 mg/kg marked decrease in the relative density of urine compared to control animals.

The results of postmortem studies have not established signs damaging effect of fullerene-(Tris-aminocaproic acid) hydrate on the body of rats after one month intramuscular injection in doses of 20 mg/kg of body weight.

Thus, the study is not installed essential features of the damaging effects of fullerene-(Tris-aminocaproic acid) hydrate on the body of rats after 30 days intramuscular injection in doses up to 20 mg/kg of the body, and at the end of the cancellation period.

Example 10. The study of the acute toxicity of fullerene-(Tris-aminocaproic acid) hydrate in laboratory animals after a single intramuscular injection.

The experiment was conducted in fgun "research center for toxicology and hygienic regulation of biopreparations" (fsri SRC TBC coatings of the FMBA of Russia), Serpukhov.

The aims of the research was to identify migrated and toxic doses, and also study the nature of the possible damaging effect on the organism of laboratory animals of the substance in a single intramuscular injection.

The experiments were carried out on outbred mice and Wistar rats from nursery GU NBMT RAMS. The animals corresponded to the sanitary rules approved by the USSR Ministry of health 06.07.73,, device, equipment is the Finance and maintenance of experimental biological clinics (vivarium). Feeding the animals ad libitum extruded feed PC-120-1 prepared according to GOST R 50258-92. Animals were quarantined and acclimatized in vivarium conditions for a period of not less than 10 days.

Experimental groups of animals were formed by random sampling with the body mass index as a leading indicator.

Solutions of the test substance in 20% aqueous DMSO solution for injection the animals were prepared under aseptic conditions ex tempora. The solution was packaged in appropriate labeled vials and before the introduction kept at a temperature of 2-6°C not more than 2 hours.

The substance was administered to mice and rats intramuscularly in doses of 5, 50 and 500 mg/kg Maximum tested dose limit maximum allowable amounts of intramuscular injection to rats and mice. Animals of the control groups were injected 20% aqueous solution in the same amount as the maximum dose of the substance.

The injected volume was adjusted to suit the individual body mass of the animal.

In the follow-up period (14 days after injection) assessed the overall condition of the animals according to their physical activity, consumption of food and water, wool and mucous membranes, body weight.

After the end of the observation period conducted postmortem of mice and rats treated with the substance at a dose of 500 mg/kg and the control substance (dissolve the spruce). The killing of animals produced by inhalation of carbon dioxide. Post-mortem examination was carried out for 1 hour after euthanasia of the animals. The morphological state of the internal organs of the animals was determined visually at autopsy.

Statistical analysis of the received results of research conducted by methods of variation statistics by student's criterion.

The results of the research. At all tested doses, the clinical symptoms of poisoning animals were absent. During the observation period, death was not, the General condition of the animal experimental and control groups did not differ. Animals willingly eating food evenly gained weight; statistically significant difference between average values of body weight of animals in experimental groups compared to the control groups were not found (table, 28).

Found that LD50substance with intramuscular injection to rats and mice of both sexes exceeds the maximum tested dose of 500 mg/kg

Postmortem mice and rats were conducted 14 days after a single intramuscular injection of the substance. Since none of the groups of animals, regardless of the administered dose of a substance, during the observation period the death was not, necropsy were subjected to only m the necks and rats, received the substance at the maximum dose of 500 mg/kg, and the animals of the control groups. The killing of animals produced by inhalation of carbon dioxide.

Physical examination of mice and rats in the experimental and control groups were noted overall picture: the coat was smooth, shiny, the leather is supple, mobile, subcutaneous tissue moderately pronounced, visible mucous membranes pale, without ulceration and foreign overlays, pathological discharges from natural orifices of the body was missing.

At postmortem autopsy also there are no differences between mice and rats of all experimental and control groups. The thoracic and abdominal cavities were anatomically correct location and normal macrostructure; any pathological changes were found. In the injection substance - thigh muscle is damaged, not detected.

Thus, the results of pathological studies have not installed signs damaging action of a substance in a single intramuscular injection to mice and rats at doses up to 500 mg/kg

Conclusion. Found that all tested doses of the substance did not cause intoxication and death in experimental animals. Values LD50fullerene-(Tris-aminocaproic acid) hydrate for mice and rats p is ivysaur maximum tested dose 500 mg/kg and, thus, exceed the maximum single therapeutic human dose of 2.9 mg/kg) in more than 170 times. Differences between species and gender sensitivity substance in doses up to 170-fold equicharacteristic not installed. The substance does not have a local irritant effect after a single intramuscular injection.

Thus, the fullerene-(Tris-aminocaproic acid) hydrate has a high therapeutic index and may not cause acute poisoning accidental overdose.

1. Hydrated N-fullerene-amino acids of General formula C60(H)3{NH(CH2)nCOOH}3·xH2O, where C60- fullerene, n=5, 6, 7, x=8 to 10.

2. A method of obtaining a compound according to claim 1, characterized in that the fullerene is subjected to interaction with a 15-fold molar excess of anhydrous potassium salts of amino acids of General formula NH2(CH2)nCOOH, where n=5, 6, 7, in the environment of aromatic solvent in slow adding to the resulting suspension phase transfer catalyst, with stirring and heating to a temperature not higher than 60-80°C until complete discoloration of the solution and formation of a solid precipitate, which represents the potassium salt obtained fullerene derivatives of amino acids, with subsequent selection, dissolved in water to obtain a 0.8 M aqueous solution, which is about srabatyvayut of 0.1 N solution of an organic or mineral acid, followed by centrifugation, washing and drying the precipitate.

3. The method according to claim 2, characterized in that the anhydrous potassium salt of amino acids used in fine condition, and the selection of sediment potassium salts of fullerene derivatives of amino acids is performed by filtering, washing with ethanol and drying.

4. The method according to any of claim 2 and 3, characterized in that as interphase catalyst is used, the methyl ether of polyethylene glycol of molecular weight of 400 or 500.

5. Pharmaceutical composition having activity against herpes virus, influenza viruses of different nature, HIV and antitumor and protivopsoriaticescoe activity, containing as active substance a compound according to claim 1 in an effective amount.



 

Same patents:

FIELD: chemistry.

SUBSTANCE: invention relates to a method of producing 1-amino-3,5-dimethyladamantane, involving reaction of 1,3-dimethyladamantane with formamide in concentrated acids to obtain 1-formamido-3,5-dimethyladamantane, provided that neither SO3-containing sulphuric acid nor 100% nitric acid is used, wherein the concentrated acids are 30-70% nitric acid and 90-100% sulphuric acid and further conversion of 1-formamido-3,5-dimethyladamantane to 1-amino-3,5-dimethyladamantane through hydrolysis with aqueous hydrochloric acid.

EFFECT: high efficiency of the method.

3 cl, 2 ex

FIELD: chemistry.

SUBSTANCE: invention relates to an ester derivative of 2-amino-bicyclo[3.1.0]hexane-2,6-dicarboxylic acid of formula (II) or the corresponding pharmaceutical salt, having group II metabotropic glutamate receptor antagonist properties. In formula X denotes fluorine, Y denotes 3,4-di-chlorobenzyloxy, R2 denotes hydrogen and R1 denotes an alkyl group selected from n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-decyl, 3-methylbutyl, 4-methylpenyl, 5-methylhexyl and 6-methylheptyl. The invention also relates to a medicinal agent and to use of formula (II) compounds as a medicinal agent for treating depression symptoms.

EFFECT: high yield.

14 cl, 4 tbl, 10 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to methods for synthesis of bicyclo[3.1.0]hexane derivatives, used as mGIuR agonists having formulae ,

, where R1 and R2 represent hydrogen, X is a halogen, R3 is -O-Ra , Ra is C1-10alkyl, and R4 is (1) hydrogen or (2) Si-(R9)(R10)(R11), where each of R9, R10 and R11 is C1-10alkyl, as well as intermediate compounds obtained when realising the said methods.

EFFECT: design of an efficient method for synthesis of bicyclo[3,1,0]hexane derivatives.

26 cl, 17 ex, 1 tbl

FIELD: chemistry.

SUBSTANCE: present invention pertains to new ester derivatives of 2-amino-bicyclo[3,1,0]hexane-2,6-dicarboxylic acid, with formula [I] or [II] and their pharmaceutical salts, with antagonistic properties towards II group metabotropic glutamate receptors. In general formulae [I] or [II] R1 and R2 are identical or different, and each stands for a C1-10alkyl group, or one of R1 or R2 is a hydrogen atom, and the other is a C1-10alkyl group, C2-10alkenyl group, C2-10alkynyl group, C1-10alkyl group, substituted with a phenyl group, possibly substituted with halogen atoms, C1-4alkyl group, C1-4 alkoxyl group; hydroxyC2-10alkyl group, halogenC1-10alkyl group, azidoC1-10alkyl, aminoC2-10alkyl, C1-10alkoxycarbonylC1-10alkyl group, farnesyl group, 4-morpholinyl-C1-10alkyl group, C1-10alkyl group, substituted with a group with formula -C(O)NRaRb (where Ra and Rb are identical or different and each represents a hydrogen atom or a C1-10alkyl group), or with a group with formula -CHRcOC(O)ZRd (where Z is an oxygen atom or a single bond; Rc represents a hydrogen atom, C1-10alkyl group or aryl group, chosen from phenyl and naphthyl, which can be substituted with a C1-4alkyl group or C1-4alkoxyl group), or substituted with a group with formula [i] [ii], (where Rd denotes the same as described above), or by a group with formula [ii]; X is a fluorine atom; and Y represents -OCHR3R4, -SR3, -S(O)nR5, -SCHR3R4, -NHCHR3R4, -N(CHR3R4)(CHR3'R4'), -NHCOR3 or -OCOR5 (where R3, R3', R4 and R4' are identical or different, and each represents a hydrogen atom, C1-10alkyl group, phenyl group, naphthyl group, phenyl group substituted with one-five substitutes, chosen from a group, consisting of a halogen atom, R5 represents a phenyl group, or a phenyl group substituted with one-five substitutes, chosen from a group, consisting of a halogen atom, and n is 1 or 2).

EFFECT: invented compounds can be used in treating and preventing mental illnesses, such as schizophrenia, anxiety and anxiety related diseases, depression, bipolar disorders and epilepsy, invention also pertains to a medicinal preparation.

30 cl, 5 tbl, 19 ex

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to derivatives of 2-amino-3-alkoxy-6-fluorobicyclo[3.10]-hexan-2,6-dicarboxylic acid and a pharmaceutical composition based on thereof possessing antagonistic effect on the group of II metabotropic glutamate receptors. Also, invention relates to a medicinal agent possessing antagonistic effect on the group II metabotropic glutamate receptors and using derivatives of 2-amino-3-alkoxy-6-fluorobicyclo[3.1.0]-hexan-2,6-dicarboxylic acid used for treatment of depressive syndromes. Invention provides enhancing effectiveness of treatment.

EFFECT: improved and valuable properties of compounds, drug and pharmaceutical composition.

19 cl, 2 dwg, 1 tbl, 15 ex

FIELD: organic chemistry, chemical technology, medicine.

SUBSTANCE: invention relates to a new method for synthesis of substituted aryl-condensed azapolycyclic compounds of formulas (II) and (VIII) , novel intermediates compounds and methods for their synthesis. Substituted aryl-condensed azapolycyclic compound of the formula (II) show the binding capacity with neurone nicotine/acetylcholine specific receptor sites and can be used for modulation of cholinergic function in treatment of different diseases. Method for synthesis of compound of the formula (II) wherein R represents hydrogen atom; R2 and R3 are chosen independently from hydrogen, halogen atom, (C1-C6)-alkyl optionally substituted with 1-3 fluorine atoms and (C1-C6)-alkoxy-group optionally substituted with 1-3 fluorine atom involves the hydrogenation reaction of compound of the formula: (1a'): synthesized from compound of the formula (III): to yield intermediate compounds of formulas (IX): and (VII): . Compound of the formula (VII) is cyclized in treatment with a base to yield compound of the formula (VIII) that is reduced. Compound of the formula (III) can be prepared by interaction of compound of the formula (IV): with compound of the formula (V): .

EFFECT: improved methods of synthesis.

14 cl, 12 sch, 64 ex

FIELD: chemistry.

SUBSTANCE: invention relates to novel diarylamines I-III of general formula , which have analgesic activity. In formula R=Ph, R1=COOMe, R2=H, R3=OH (I); R=n-FC6H4, R1=COOMe, R2=H, R3=OH (II); R=Ph, R1=COOMe, R2=Me, R3=OH (III).

EFFECT: invention also relates to a method of obtaining said diarylamines.

2 cl, 1 tbl, 4 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to versions of the method of producing a derivative of tetrafluorobenzyl-5- aminosalicylic acid of formula I where R1, R2 and R3 can independently denote hydrogen or halogen, which can be used to prevent or treat acute and chronic neurodegenerative diseases, particularly focal brain ischemia, and to a method of producing pharmaceutically acceptable salts of this derivative. One version of the method of producing the tetrafluorobenzyl-5-aminosalicylic acid derivative of formula I involves the following steps: a) oxidation of tetrafluorobenzyl alcohol of formula I to tetrafluorobenzaldehyde of formula 2 b) conversion of tetrafluorobenzaldehyde to a tetrafluorobenzylidine-5-aminosalicylic acid derivative of formula II through a dehydration-condensation reaction between tetrafluorobenzaldehyde and 5-aminosalicylic acid of formula 3 and c) hydrogenation of the tetrafluorobenzylidine-5-aminosalicylic acid derivative to a tetrafluorobenzyl-5-aminosalicylic acid derivative of formula I.

EFFECT: efficient method of producing tetrafluorobenzyl-5-aminosalicylic acid derivative.

4 cl, 9 tbl, 1 dwg, 11 ex

FIELD: chemistry.

SUBSTANCE: invention refers to acid addition salts of 5-amonilevulinic acid ester (5-ALA ester) with acid in the form of solphonic acid derivative selected out of C1-C4-alkanesulphonic acid, benzolsulpnonic acid substituted with C1-4-alkyl, 2-hydroxy-ethanesulphonic acid and (+)-camphor-10-sulphonic acid, or nitric acid, where 5-ALA ester is a compound of the formula R22N-CH2COCH2-CH2CO-OR1 (R1 is unramified or ramified C1-6-alkyl group which can be possibly interrupted with one or two -O- groups and possibly substituted with phenyl, which in turn can be substituted with unramified or ramified C1-6-alkyl group; R2 is hydrogen atom). Also invention refers to methods of obtainment of the claimed salts, their application, pharmaceutical composition and methods of photochemotherapeutical treatment and in vitro diagnostics.

EFFECT: application of claimed salts as photosensitising agents in photodynamic therapy or diagnostics.

23 cl, 2 tbl, 10 dwg, 42 ex

FIELD: chemistry.

SUBSTANCE: method of obtaining compound of the formula VIII where R12 and R13 each are removable protective group where protective group R12 is more resistant to extraction by hydrolysis or hydration than R13. Method involves reaction of compound of the formula IX with relevant R12 donor compound selected out of anhydrides, halogenides, carbamates and N-hydroxysuccinimides.

EFFECT: improved method.

4 cl, 6 ex

FIELD: organic chemistry, chemical technology.

SUBSTANCE: invention relates to novel 1,2,3-tris-[(ammonio)methylcarbonyloxypoly(alkyleneoxy)]propane trichlorides of the formula: wherein: if R1 = R2 = R3 mean -CH2CH2OH then a + c + e (the total degree of oxypropylation) = 49, b + d + f (the total degree of oxyethylation) = 9; if R1 = R2 = R3 mean -CH2CH2OH then a + c + e = 55, b + d + f = 10; if R1 = R2 = R3 mean -CH2CH2OH then a + c + e = 66, b + d + f = 15; if R1 = R = R3 -CH2CH2OH then a + c + e = 76, b + d + f = 18; if R1 = R2 mean hydrogen atom (H); R3 means aliphatic hydrocarbon radical comprising 10-16 carbon atoms then a + c + e = 76, b + d + f =18; if R1 = R2 mean H; R3 means aliphatic hydrocarbon radical comprising 17-20 carbon atoms then a + c + e = 49, b + d + f = 0; if R1 = R2 mean H; R3 means aliphatic hydrocarbon radical comprising 17-20 carbon atoms then a + c + e = 55, b + d + f = 0; if R = R means H; R3 means then a + c + e = 49, b + d + f = 9; if R1 = R2 mean H; R3 means then a + c + e = 55, b + d + f =10, and to a method for their synthesis. Method involves interaction 1,2,3-tris-[hydroxypoly(alkyleneoxy)propanes of the formula: wherein a + c + e = 49-76, b + d + f = 0-18 with monochloroacetic acid in the presence of acidic catalyst, in boiling organic solvent medium and azeotropic removal of formed water and the following treatment of the synthesized reaction product at heating with amino-compounds of the formula: R1R2N wherein R1 = R2 mean -H, -CH2CH2OH; R3 means -CH2CH2OH, aliphatic radical comprising 10-16 or 17-20 carbon atoms, and in the molar ratio of reagents - propane hydroxyl derivatives : monochloroacetic acid : amino-compounds = 1:(3.0-3.2):(3.0-3.2), respectively. Novel compounds possess properties of emulsifiers of aqueous-mazut emulsions.

EFFECT: improved method of synthesis, valuable technical properties of compounds.

6 cl, 1 tbl, 10 ex

FIELD: organic chemistry, chemical technology.

SUBSTANCE: invention relates to novel 1,2,3-tris-{[aminopoly(ethyleneamino)ethylammonio]-methylcarbonyloxypoly(alkyleneoxy)]}propane trichlorides of the formula:

wherein at a + c + e (the general degree of oxypropylation) = 49; b + d + f (the general degree of ethylation) = 0, n = 1-6; at a + c + e = 55, b + d + f = 0, n = 1-6; at a + c + e = 49, b + d + f = 9, n = 1-6; at a + c + e = 10, b + d + f = 10, n = 1-6; at a + c + e = 66, b + d + f = 15, n = 1-6; at a + c + e = 76, b + d + f = 18, n = 1-6, and to a method for their synthesis. Method involves interaction of 1,2,3-tris-[hydroxypoly(alkyleneoxy)]propanes of the formula:

wherein a + c + e = 49-76, b + d + f = 0-18 with monochloroacetic acid in the presence of acidic catalysts, in boiling organic solvent medium, azeotropic removing water formed and the following treatment at heating the synthesized reaction product with polyethylenepolyamines of the formula: H2N(CH2CH2NH)nCH2CH2NH2 wherein n = 1-6, and in the following mole ratios of reagents - hydroxyl derivatives of propane : monochloroacetic acid : polyethylenepolyamines = 1:(3.0-3.2):(3.0-3.2), respectively. New compounds possess the fungicide activity, properties of emulsifiers of cationic bitumen emulsions, capacity to enhance adhesion of bitumen to mineral materials.

EFFECT: improved preparing method, valuable properties of compounds.

6 cl, 3 tbl, 6 ex

FIELD: organic chemistry, chemical technology.

SUBSTANCE: invention relates to novel 1,2,3-tris-[(ammonio)methylcarbonyloxypoly(alkyleneoxy)]-propane trichlorides of the general formula:

wherein at -X+ as -N+R1RR, R1 = R2 mean hydrogen atom (H), R3 means aliphatic hydrocarbon radical comprising 10-16 carbon atoms, a + c + e (the general degree of oxypropylation) = 49,b + d + f (the general degree of oxyethylation) = 0; at -X+ as -N+R1R2R3, R1 = R2 mean H, R3 means aliphatic hydrocarbon radical comprising 10-16 carbon atoms, a + c + e = 55, b + d + f = 0; at -X+ as -N+R1R2R3, R1 = R2 mean H, R3 means aliphatic hydrocarbon radical comprising 10-16 carbon atoms, a + c + e = 80, b + d + f = 24; at -X+ as -N+R1R2R3, R1 = R2 mean H, R3 means aliphatic hydrocarbon radical comprising 10-16 carbon atoms, a + c + e = 90, b + d + f = 27; at -X+ as -N+R1R2R3, R1 = R2 means H, R3 means phenyl, a + c + e = 80, b + d + f = 24; at -X+ as -N+R1R2R3, R1 = R2 mean H, R3 means phenyl, a + c + e = 90, b + d + f = 27; at -X+ as , a + c + e = 80, b + d + f = 24; at -X+ as , a + c + e = 90, b + d + f =27. Also, invention relates to a method for synthesis of these compounds. Method involves interaction of 1,2,3-tris-[hydroxypoly(alkyleneoxy)]-propane of the formula:

wherein a + c + e = 49-90, b + d + f = 0-27 with monochloroacetic acid in the presence of acidic catalyst, in boiling organic solvent medium with azeotropic removal of water formed and the following treatment of synthesized reaction product in polar solvent medium at heating with amino-compounds of the formula: NR1R2R3 wherein R1 = R2 mean H, R3 means aliphatic hydrocarbon radical comprising 10-16 carbon atoms, phenyl, or morpholine of the formula:

in the following mole ratios of reagents - propane hydroxyl derivative : monochloroacetic acid : amino-compound or morpholine = 1:(3.0-3.2):(3.0-3.2), respectively. New compounds show the bactericidal and fungicide activity and properties of demulsifying agents for petroleum emulsions.

EFFECT: improved method of synthesis, valuable properties of compounds.

7 cl, 3 tbl, 8 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely oncology and radiology, and may be used for treating stage IIA Hodgkin's lymphoma. That is ensured by 3-4 therapeutic courses of chemotherapy by the scheme ABVD - adriablastine, bleomycine, vinblastine, dacarbazine. It is followed by chemotherapy by the scheme AVB - adriablastine, vinblastine, dacarbazine. From the 2nd day of the AVD course, the radiation therapy covering initially involved zones is applied at single basic dose 1.2 Gy, daily 2 times a day every 4 hours. If the initial tumour lesion is less than 5 cm, the radiation therapy is applied to total basic dose 30 Gy, while the lesions more than 5 cm requiring dose up to 36 Gy.

EFFECT: method provides reducing total treatment length at least by 2 months, reducing a risk of recurrences, reducing a probability of secondary tumours and late complications of the radiation therapy ensured by the intensified combined chemoradiation treatment.

2 ex

FIELD: medicine.

SUBSTANCE: group of inventions refers to versions of applying an antisecretory protein which corresponds the amino acid sequence SEQ ID NO:6, its homologue, and/or a fragment containing the amino acid sequence SEQ ID NO:4 showing equivalent activity, and/or its pharmaceutically active salt for preparing a pharmaceutical composition and/or nutritional care for treatment and/or prevention a dysfunction, e.g. a pathological function, lipid raft, receptor and/or small pit hypo- or hyperfunction. The lipid raft, receptor and/or small pit dysfunction can be induced by or cause a number of the other conditions, such as vascular and pulmonary dysfunctions and/or endocrine disorders, e.g. diabetes and related disorders.

EFFECT: group of inventions enables controlling intracellular transport and cell product release, as well as normalising tissue component distribution in various diseases.

13 cl, 12 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to oncology and deals with method of treatment in case of urinary bladder cancer. Combined treatment with application of neoadjuvant system polychemical therapy by the following scheme: 1 day -5-fluoruracyl, 2 day - cyclophosphane+vinblastine, 14 day - methotrexate, 15 day - doxorubicin, 16 day -cisplatin, and adjuvant intrabladder chemical therapy with preparation doxorubicin during 2 years, on the first day after operation doxorubicin 40 mg is instilled, then 4 instillations of doxorubicin in dose 40 mg with 7 day interval, then doxorubicin 40 mg is instilled with 30 day interval during 11 months, after that during the second year instillations of doxorubicin in dose 20 mg are performed after 1 month.

EFFECT: invention ensures improvement of results of organ-preserving treatment by reduction of number of urinary bladder cancer recurrences.

1 ex

FIELD: medicine.

SUBSTANCE: there are offered: recovered polynucleotides and polypeptides for binding human EGFR as a part of an antibody, a vector and a host cell for antibody expression, a method for producing an anti-EGFR antibody and an antibody fragment, the anti-EGFR antibody and the antibody fragment. There are discussed a composition containing the anti-EGFR antibody or its fragment, as well as applying the antibody and its fragment for treating EGFR-associated disorders. Besides, there are described versions of glycosylation of the anti-EGFR antibody or its fragment.

EFFECT: invention can find further application in therapy of various EGFR-mediated diseases.

30 cl, 6 ex, 32 dwg, 39 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to cytotoxic compounds of directed action and methods for therapeutic use thereof in treating neoplasm and other diseases.

EFFECT: high treatment efficiency.

9 cl, 12 tbl, 31 ex, 3 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to {1-methyl-5-[2-(5-trifluoromethyl-1H-imidazol-2-yl)pyridin-4-yloxy]-1H-benzimidazol-2-yl}(4-trifluoromethylphenyl)amine salt. Also, the invention refers to a pharmaceutical composition based on the declared salt, to a method for preparing said pharmaceutical composition and to a method of treating cancer and/or angiogenesis based on the use of the declared salt.

EFFECT: there are prepared new {1-methyl-5-[2-(5-trifluoromethyl-1H-imidazol-2-yl)pyridine-4-yloxy]-1H-benzimidazol-2-yl}(4-trifluoromethylphenyl)amine salts showing effective biological properties.

42 cl, 12 dwg, 10 tbl, 7 ex

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