Application of polysaccharide derivatives as anti-infection substance

FIELD: medicine; pharmacology.

SUBSTANCE: invention claims application of starches substituted by quarterly ammonium groups with substitution degree of 0.4 to 3.0 with the help of a linker, in infection diseases treatment. Suppression of 1 type herpes virus replication by the claimed starches is demonstrated, in comparison to zero antivirus effect of non-modified starches. Minimal inhibiting concentration for staphylococci and mycobacteria is 5-60 g/l.

EFFECT: efficient infection disease treatment by the claimed starches.

5 cl, 7 tbl

 

The invention relates to the use of substances on the basis of oligo - and polysaccharides as anti-infective tool, such as an antibacterial and antiviral agent. This anti-infective agent may be applied, for example, in cosmetic and pharmaceutical formulations as preservatives in pharmaceutical formulations as biologically active compounds, for the biocidal treatment of surfaces, fabrics and packaging materials, for example, for food products or products used in medicine, biology, pharmacy, and a protective coating RAS for use in cosmetic and pharmaceutical industries, in agriculture, in the food industry and in the production of animal feed.

It is known that globally increased infection with bacterial pathogens and antimicrobial resistance is a major public health problem. The worldwide increase in TB infections mycobacterial strains, acquiring over time resistance to conventional therapeutic agents (see ..Bloom, J.L.Murray, Science 257, 1992, 1055), and therapy of infections with multiresistant staphylococci (.Kresken, Bundesgesundheitsblatt 38, 1996, 170), requires the development of new biologically active substances. Vital alternative biologically and is active substances with new mechanisms of action, especially in relation to resistance to antibiotics and treatment of bacterial infections in intolerance against existing biologically active substances.

With the development of highly selective nucleoside and nucleotide virustatic, such as, for example, acyclovir, penciclovir, ganciclovir, sorivudine and cidofovir, herpes virus, although progress has been made in the fight against life-threatening infection, however, called therapeutic agent are one and the same principle. They inhibit viral DNA polymerase. Another disadvantage of these compounds is that they are also involved in the metabolism of DNA of infected cells and therefore pose a danger to cause mutagenic, teratogenic and carcinogenic effects (Wutzler, P.Thust, R.Antiv. Res. 49, 2001, 55). In addition, if long-term use of nucleoside and nucleotide virustatic as in infected cell cultures and in patients with reduced immunity proved the development of resistance to these drugs (Andrei, G. et al., Antimicrob. Agents Chemother., 1995, 39 1632; Pavic, J. et al., Antimicrob. Agents Chemother., 1997, 39 2686). Therefore, should be further developed new highly-active anti-virus prophylactic and therapeutic agent with a different mechanism of action.

A large group of biologically active compounds are the Quaternary and monavie connection. They can destroy microorganisms, such as bacteria and fungal organisms. Low molecular weight Quaternary ammonium compound used as a disinfectant or biocide coatings (J.Controlled Release 50, 1998, 145). A typical problem of low molecular weight compounds is insufficient manifestation of biological activity, for example, as a result of various processes of migration and splitting. Polymeric Quaternary amidofunctional substances can be synthesized from commercially available Quaternary ion exchange resins graptoloidea polyurethanes with hydroxyalkylated polybutadiene or polysiloxanes with primary alcohol functional groups in the side chain. These biocidal polymers are often high cost of production and often toxic because they contain residues of toxic monomers (Trends in Polymer Science 4, 1996, 364). In addition, may be undesirable and dangerous accumulation of polymer in the body, as they do not decompose biologically. In addition, synthetic polymers containing cationic functional group, is used in the form of dispersions for wood protection (U.S. patent US 5049383). The disadvantage of synthetic polymers containing cationic functional group, is a high-cost production, toxicity (contamination of the residual content of monomers) and resistant to biological decomposition.

Derivatives of polysaccharides with Quaternary ammonium functional groups are known and are still used primarily as additives to improve the properties of the surface in the paper and textile industries, as well as in cosmetics as a consistency regulator, and they have only a low degree of substitution (DS)of less than 0.2. Their biological activity is still not known. On the other hand, describes the anti-infective action, especially antibacterial action, ethers of starch, which contain long alkyl chain with 8-22 carbon atoms and is linked to the starch via a silyl ether group (patent application Japan JP 05295002). Low chemical stability alkylsilanes simple ether polysaccharides leads to uncontrolled release of functional groups even when exposed to moisture in the air and result in the reduction or loss of biological activity (D.Klemm et al., Comprehensive Cellulose Chemistry, Wiley-VCH, 1998). In addition, low molecular weight silyl compound toxic. In addition, refer to publications about antibacterial cellulose fibers and derivatives of chitosan (W.H.Daly, M.M.Guerrini, Polym. Mat. Sci. Eng. 79, 1998, 220). Chitosan is most often described as a natural cationic polysaccharide and is used as a fungicidal agent in cosmetics .Tashiro, Macromol. Mater. Eng. 286, 2001, 63, ..Gupta, M.N.V.R. Kumar, J.M.S.-Rev. Macromol. Chem. Phys. C40, 2000, 273). The disadvantages of this polysaccharide are frequent contamination of other nutrients, high costs due to expensive methods of isolation and purification and natural structure, confining ammonium group solely on the main polymer chain. In addition, their distribution is irregular, and the content is limited by the degree of substitution of 1. Also described superabsorber of cation modified and cross-linked polysaccharides, such as cellulose (European patent application EP 0582624 B1).

In the literature there is evidence that biological activity is determined by the presence of Quaternary ammonium groups, and, on the other hand, describes what a typical connection with a cationic Quaternary alkyl substituted amino groups, such as Polyquaternium 10 proven not possess bioactivity (W.A.Daly, M.M.Guerrini, D.Culberson, J.Macossay, Science and Technology of Polymers and Advanced Materials, Plenum Press, 1998, 493). From the available literature information in any way it is impossible to conclude whether such patterns are actually biologically active.

The basis of the invention is to find polymers with new anti-infective action, and the matter should have an effective anti-infective broad spectrum activity, to allow effective the active combating resistance to antibiotics in bacterial infections, to provide new opportunities for the treatment of viral infections, as well as to be well-tolerated, biodegradable, non-toxic and produced by a simple method.

According to the invention the task is solved by obtaining the derivatives of polysaccharides, which by the esterification reaction with the formation of ethers entered cationic functional group with a degree of substitution (DS) in the range from 0.4 to 3.0, in particular alkylammonium group with a degree of substitution in the range from 0.6 to 1.8 through spacer elements of the group. Polysaccharides are particularly suitable due to their biological decomposing and nontoxicity.

In accordance with this invention relates to the use of polysaccharides, substituted connected by linker Quaternary ammonium groups with a degree of substitution of from 0.4 to 3.0, as anti-infective funds or for the treatment of infectious diseases.

Used according to the invention compounds have high biological activity and in an unexpected way inhibit the growth of pathogenic bacteria, such as staphylococci and bacilli, with a minimum inhibitory concentration in the range 5-60 mg/l, and also inhibit the replication of herpes viruses and influenza at a concentration in the range of 3-50 mg/L. Due to its connection properties can use the I for the manufacture of medicines, designed to prevent and treat bacterial and viral infections. They can be used both independently and in combination with known therapies or physiologically compatible auxiliary substances and substances-carriers.

Anti-infective compounds can be prepared for use as solution or suspension in a pharmaceutically acceptable environments for local or parenteral application by intravenous, subcutaneous or intramuscular injection, for vnutripuzarnogo application; as tablets, capsules or suppositories. Connections may be used in doses of 0.1-1000 mg/kg of body weight.

To obtain biologically active compounds according to the invention using polysaccharides, preferably polyglycine, such as cellulose, Lichonin, pullulan, dextran, and especially preferably starches, such as natural starches of different origin, such as potato, wheat, maize and rice starch, and chemically or enzymatically partially hydrolyzed starches, such as solami (Solamy), amylose, amylopectin and starch, waxy corn and starches derived from genetically modified plants, such as starch brand Hylon. The content of amylose and amylopectin in the starch composition is yet respectively from 0 to 100%, preferably 30-70%. The molecular weight of suitable polysaccharide is in the range of 103-107g/mol (see Table 1). Anhydroglucose link - repeatability units (AGU) can be schematically depicted through communication: α(1-4), α(1-6), α(1-3), β(1-4) and β(1-3) or through a combination of both, as, for example, α(1-4) and α(1-6) (see the structure of formula (A))or α(1-6), α(1-3) and α(1-4) can be connected to each other and contain different lengths and associated lateral links. In addition, may contain other functional groups, such as difficult ether phosphate group, for example, natural potato starch.

Patterns used polysaccharides can be represented, for example, by formula (A).

Table 1
StarchesThe content of amylose (%)Molecular weight (GPC1), (g·mol-1)
Starch brand Hylon VII (H)709·106
Potato starch (P, Emsland)2840·106
Maize starch (M)2876·106
Wheat starch (W)26 65·106
starch is waxy maize (WM)151·106
Solamyl (S)289700
GPC1gel-pronikayushie chromatography; defined in dimethyl sulfoxide..

The degree of interaction between hydroxyl groups is characterized by an average degree of substitution (DS). This average determines without any differentiation, the number of functional hydroxyl groups and, therefore, he is called polysaccharides according to the definition in the range from 0 to 3. The degree of substitution (DS) of cationic groups derived polysaccharides with anti-infective activity according to the invention is in the range between 0.4 and 3.0, preferably between 0.6 and a 1.8. In the case of the introduction in the process of obtaining derivatives of functional groups, which themselves contain reactive groups, for example hydroxyl groups due to the formation of ethers of polysaccharides with epoxides, such groups can also react with the formation of longer side chains.

Used according to the invention derivatives of polysaccharides are known or can be obtained in a known manner, in particular through the formation of ethers of polysaccharides with reactive compounds and, resulting in either formed directly Quaternary ammonium compounds of General formula (I) (PS: radical polysaccharide, found only one Deputy)or the quaternization is carried out directly after the formation of simple ether. In the formula (I) R1, R2and R3independently from each other may be preferably an alkyl with 1-4 carbon atoms or a benzyl or substituted benzyl (deputies, for example, may be present in an amount of from 1 to 3 and represent alkyl, halogen, alkoxy, carbarnoyl, alkoxycarbonyl, cyano, dialkylamino). R1can also be hydrogen, and X is an anion (for example, a halide, a hydroxyl group, sulfate, hydrosulfate, and other anion mineral and carboxylic acids), n can mean 2-4.

Preferably used reagents, leading to the formation of Quaternary cationic groups, are 2,3-epoxypropyltrimethylammonium (QUAB®151, Degussa AG, formula (II) or 3-chloro-2-hydroxypropyltrimethylammonium (QUAB®188, Degussa AG, formula (III). For the formation of ethers of polysaccharides can also be used reagents of General formula IV, where Y represents chlorine, bromine, and n is 1-3.

In line with this we are talking about the linker, what exploits which Quaternary ammonium groups associated with the polysaccharide, about optionally substituted by hydroxyl alkylene with 2-4 carbon atoms.

The formation of ethers to obtain biologically active derivatives of polysaccharides can be performed in various ways and implemented in a known way, and achieved a high content of cationic groups. Suitable suspension of the polymer in alcohol and caustic sodium and water (heterogeneous way), and as a reaction medium suitable alcohols, such as methanol, isopropanol, preferably ethanol, or aqueous alkaline solutions, preferably caustic sodium/water with the transition from a homogeneous system for the heterogeneous and homogeneous solutions of polymers in bipolar aprotic solvents such as dimethyl sulfoxide or dimethylacetamide, in the presence of lithium chloride or other solvents. Interact with cationizing reagent is in the range between 1 and 48 hours, preferably from 3 to 24 hours, and the temperature is between 30 and 130°C, preferably from 40 to 80°C. Through the use of a molar equivalent of the agent used for the formation of simple ether, you can further adjust the degree of product substitution and to vary it within wide limits. To obtain the derivatives of polysaccharides suitable, in addition, a multi-stage reaction, when already canonized amino is functionalliteracy the product is again subjected to interaction in the above-mentioned conditions. Processing of the reaction product is carried out in the usual way chemistry of polymers, and low molecular weight by-products and residues of reagents separated by dialysis or washing or presidenial of water in an organic solvent.

The degree of substitution calculated found by elemental analysis average nitrogen rate according to the following formula:

In addition, to determine the degree of substitution (DS) is suitable contained in the compounds counterions such as chloride, and NMR spectra.

The following embodiments serve more detailed explanation of the invention, but they should not limit the invention.

Examples of carrying out the invention

1. The formation of compounds by heterogeneous reactions in the environment of the ethanol/sodium hydroxide/water

20 g of the polysaccharide (polysaccharide see Table 2) are suspended in 80 ml of ethanol. To this suspension is added dropwise a solution 10,85 g of sodium hydroxide in 28 ml of water and 80 ml of ethanol, and the solution 0,246 mol QUAB®188 (69%aqueous solution). The reaction mixture was stirred at 60°C for 6 hours. The product is neutralized HCl 0.1 N., subjected to dialysis and dried by freezing.

The product yield was 95% (achieved a degree of substitution).

Elemental analysis: N % 3,51.

The average degree of samadani the (DS N)=0,66

Table 2

Heterogeneous cationisation
ReagentsThe degree of substitution
PolysaccharideTypeThe molar ratio of AGU:reagentProduct
HylonQUAB®1881:3H1(466)0,50
HylonQUAB®1881:2H20,66
(KS005)
AmicaQUAB®1881:3WM10,14
-(505)
Potato starchQUAB®1881:3P11(491)0,34
Potato starchQUAB®1881:3P21(469)0,58
Wheat starchQUAB®1881:3,25W1(527)0,99
Wheat starchQUAB®1881:2W3(571)Wheat starch21:5W40,72
Wheat starch21:10W50,81
1different concentration of sodium hydroxide

2reagent: Cl-CH2-CH2-N(C2H5)H2Cl

2. The interaction of polysaccharides in aqueous solution of sodium hydroxide

20 g of the polysaccharide (polysaccharide see Table 3) are suspended in a solution of sodium hydroxide (0.5 g of sodium hydroxide in 100 ml of water). The mixture was stirred at 60°C. At this temperature, added dropwise 0,123 mol QUAB®151, or in the case of the split starch Solamyl added dropwise QUAB®188. The reaction mixture was stirred at 60°C for 6 hours. After cooling to room temperature, the mixture is neutralized HCl 0.1 N.; then subjected to dialysis and dried by freezing.

The product yield was 98% (achieved a degree of substitution).

Elemental analysis: N % 3,34.

The average degree of substitution (DSN)=0,60.

Table 3

Hetero-homogeneous cationisation
ReagentsThe degree of substitution
Polysaccharide TypeThe molar ratio of AGU: reagentProduct
HylonQUAB®1511:1H3(477)0,40
HylonQUAB®1511:2H4(KS006)0,92
AmicaQUAB®1511:0,5WM2(503)0,38
AmicaQUAB®1511:1WM3(501)0,60
AmicaQUAB®1511:2WM4(502)0,92
Maize starchQUAB®1511:0,5M1(517)0,35
Maize starchQUAB®1511:1M21(516)0,55
Maize starchQUAB®1511:1M31(519)0,72
Maize starchQUAB®1511:2M4(515)1,03
Potato starchQUAB®1511:3P3(KS016)0,69
Potato starchQUAB®1511:2P4(K013) 1,05
Wheat starchQUAB®1511:0,5W4 (520)0,39
SolamylQUAB®1881:2S11(554)0,68
SolamylQUAB®1881:3S2(555)0,76
SolamylQUAB®1881:2S31(568)0,80
1different concentration of sodium hydroxide

3. Homogeneous reaction in dimethyl sulfoxide (DMSO)

15 g of the polysaccharide (polysaccharide see Table 4) are suspended in dimethyl sulfoxide (DMSO) at room temperature and heated to 80°C, while the polysaccharide is dissolved. The solution is cooled to room temperature and add 0.5 g of sodium hydroxide dissolved in 20 ml of water. Then under stirring was added dropwise 0,0925 mol QUAB®151. The reaction mixture was stirred at 60°C for 24 hours. After cooling to room temperature, the mixture is neutralized HCl 0.1 N., and then subjected to dialysis and dried by freezing.

The product yield was 99% (achieved a degree of substitution).

Elemental analysis: N % 3,22.

The average degree of substitution (DSN)=0,57

Table 4

Homogeneous cationisation
ReagentsThe degree of substitution
PolysaccharideTypeMolar with respect AGU:reagentProduct
HylonQUAB®1511:3N(436)0,55
Potato starchQUAB®1511:1P5(KS9)0,42
AmicaQUAB®1511:1WM5(504)0,57
Wheat starchQUAB®1511:1W5(531)0,41
Maize starchQUAB®1511:1M5(532)0,40
SolamylQUAB®1511:1S4(588)0,60

4. Cationisation in several stages

5 g cationizing polysaccharide (polysaccharide see Table 5) are suspended in a solution of sodium hydroxide (0.5 g of sodium hydroxide in 100 ml of water). The mixture was stirred at 60°C for one hour. At this temperature, was added dropwise 0.2 mol QUAB®151. The reaction mixture was stirred at 60°is within 6 hours. After cooling to room temperature, the mixture is neutralized HCl 0.1 N., and then subjected to dialysis and dried by freezing.

The product yield was 95% (achieved a degree of substitution).

Elemental analysis: N % 5,11.

The average degree of substitution (DSN)=1,32

Table 5

Cationisation in several stages
Cationically starchReagentThe degree of substitution DSN
PolysaccharideThe original DSTypeThe molar ratio of AGU:reagentProduct
Hylon0,40QUAB®1511:2N6(479)0,90
Hylon0,80QUAB®1511:1,5H7(498)1,10
Potato starch0,77QUAB®1511:2P6(506)1,18
Potato starch0,42QUAB®1511:10P7(597)1,32
Amica0,60QUAB®1511:10WM6(598)1,25
Wheat starch0,41QUAB®1511:10W6(603)1,16
Wheat starch0,88QUAB®1511:10W7(602)1,41
Maize starch0,55QUAB®1511:10M6(600)1,13
Potato starch1,32QUAB®1511:15R81,80
Maize starch1,13QUAB®1511:10M71,65
Solamyl0,80QUAB®1511:2S5(610)0,91

5. Determination of the antibacterial/antimicrobial activity of compounds against gram-positive and gram-negative bacteria and yeast

Antibacterial activity of compounds against Staphylococcus aureus SG 511, S.aureus 134/93 (politicise) and Mycobacterium vaccae IMET 10670 was tested by determining the minimum inhibitory concentration (MIC) test breeding in microbalance Mueller-Hinton (DIFCO) in accordance with the recommendations of NCCLS [National Committee for Clinical Laboratory Standards: Methods for dilution antim-icrobial susceptibility tests for bacteria that grow aerobically;4 thEd.; Villanova, Ed.; Approved standard Document M7-A4. NCCLS, (1997)]. The results are presented in Table 6.

Table 6

Antibacterial activity
SampleThe degree of substitution DSNMIC [mg/l]
Staphylococcus aureusMycobacterium vaccae IMET 10670
SG511134/93
H4 KS0060,92the 15.6the 15.67,8
H7 4981,10the 15.6the 15.67,8
WM 4 5020,9231,2562,5the 15.6
M3 5190,72the 15.662,57,8
M4 5151,0331,2562,5the 15.6
P3 0160,6931,2562,57,8
P4 0131,05the 15.6125a 3.9
W2 5670,5762,562,5the 15.6
W3 5710,6131,2531,257,8
W1 5270,9931,25 31,25the 15.6
S1 5540,6831,2562,6the 15.6
S2 5550,76the 15.631,257,8
S3 5680,80the 15.6the 15.67,8

6. Determination of the antiviral action against herpes simplex virus type 1 (HSV-1)

Before testing the antiviral activity was determined in 50%cytotoxic concentration (CC50in the cells of green monkey kidney (KAP) in order to exclude non-specific action of the substance. For this cell KAP seeded in microtiter plate with a series of diluted substances (factor 2) (Schmidtke et al.; J.Virol Meth. 95, 2001, 133). After 72 hours incubation was carried out by staining the cells with a solution of the dye crystal violet in methanol. After separation of the dye solution, measure the optical density of the individual recesses in a plate photometer company Dynatech (550/630 nm) and compared with an average of six untreated control samples of cells, which is taken as 100%. CC50represents the concentration of a substance at the point of intersection of the curve of extinction series of dilutions from 50%line average control. Antiviral activity of compounds against HSV-1 and is having experienced in the test for cytopathic effect ingibirovaniya (zpE-Hemmtest) in the cells of the kidney green monkey and was identified in 50%dose inhibition (IC 50) (Schmidtke et al.; J.Virol Meth. 95, 2001, 133). Expected index of selectivity as the ratio of the SS50and IC50(Table 7). The parent compound (QUAB-reagents or unmodified starches) did not show any antiviral activity (results not presented).

Table 7

Antiviral activity
SampleThe degree of substitution DSNSS50(ág/ml) in the KAP-cellsIC50(µg/ml) against HSV-1Index selectivity (CC50/IC50)
H3 4770,40more than 2008,54more 23,42
H1 4660,50more than 2005,11more 39,14
H2 0050,66more than 2007,07more 28,29
WM 2 5030,38more than 2007,05more 28,37
M1 5170,35more than 200to 7.84more 25,51
P1 4910,34more than 20010,15more 19,70
W4 5200,39more than 20010,55 more 18,96
S1 5540,68141,543,5939,43

7. Determination of mechanism of action in a modified test to determine the reduction spots (plaque reduction test)

Investigation of the mechanism of action of the substance were sensitive to acyclovir and phosphonopropionic acid HSV-1 strain Kupka in a modified test to determine the reduction of spots, for example, with connection M 1 (Schmidtke et al.; J.Virol Meth. 95, 2001, 133). When this was carried out by adding substances in various concentrations:

1. only it does not contain cells with the virus (106pfu/ml), which is then incubated with the compound at a temperature of 37°C for 6 hours and used after dilution of the substance in the test to determine the reduction spots: no decrease of platelets in doses of 6.25-25 μg/ml (results not shown);

2. only in agar: no decrease of platelets in doses of 6.25-25 μg/ml (results not shown);

3. only during the one-hour adsorption at 4°C for 2 hours (3,12-12.5 µg/ml) and

4. 1,2 and 4 hours before the addition of virus (3,12-12.5 µg/ml).

The results of studies of the mechanism of action show that the compounds according to the invention have no effect on the virus, as inaktivirovanie not containing cells of the virus could not be installed. So the m for inhibition of the replication (reproduction) of the herpes virus is rather, the presence of substances before or during the adsorption of the virus on the test cells.

1. The use of starches, substituted attached through a linker Quaternary ammonium groups with a degree of substitution of from 0.4 to 3.0, for the treatment of infectious diseases.

2. The use of starches according to claim 1, whereby the degree of substitution is from 0.6 to 1.8.

3. The use of starches according to claim 1, and represent substances such as potato, wheat, maize and rice starches, chemically or enzymatically partially hydrolyzed starch or starch derived from genetically modified plants.

4. The use of starches according to claim 1, wherein the linkers through which the Quaternary ammonium groups linked polysaccharide, are optionally substituted hydroxyl group alkylene group with 2-4 carbon atoms.

5. The use of starches according to claim 1 for the treatment of viral and bacterial infectious diseases.



 

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Poplar-aspen oil // 2326685

FIELD: medicine; pharmacology.

SUBSTANCE: agent contains oil extract of poplar buds and sprouts and oil extract of aspen buds and sprouts with component ratio as follows, mass.%: oil extract of poplar buds and sprouts 20-90, oil extract of aspen buds and sprouts 10-80, mixed oil extract of poplar buds and sprouts and oil extract of aspen buds and sprouts, with component ratio as follows, mass.%: oil extract of poplar buds and sprouts 20-90, oil extract of aspen buds and sprouts 10-80, as well as fat extract of plant raw with plant component ratio as follows, mass.: marsh tea 3, comfrey 3, blueash 2, silver fir 1, common burdock (root) 2, horseheal (root) 1, deer's-tongue 1, horseradish 1, musquash-poison 2, hog bean (herb) 2, hop (cones) 2, cowberry (root) 2, mountain arnica (blossom) 2, ratio of oil extracts and fat extract of plant raw is 1:3 respectively.

EFFECT: agent allows widening range of preventive and therapeutic herbal medicinal agents of antiviral and anti-inflammatory action.

2 cl, 9 ex

FIELD: veterinary; veterinarian virology.

SUBSTANCE: production of dry cultural rinderpest virus-vaccine for minor ruminants includes growing of virus containing raw materials from "45G37/35-K" rinderpest virus strain in cell culture, introduction of protective medium and production of the end product. Passaged cell culture of saiga kidney is used as a cell culture. Infected culture is incubated under roller conditions during 5-7 days with the change of supporting medium each 2-3 days. Virus containing culture is mixed before freeze drying in proportion 1:1, and then freeze dried until moisture mass fraction is no more than 4%. Finally, it is packed into ampoules. The end product contains virus raw materials, peptone, sucrose, gelatine and demineralised water.

EFFECT: high-performance production of standard and innocuous rinderpest virus-vaccine for minor ruminants stable for storage conditions.

2 cl, 3 ex

FIELD: medicine, pharmacology.

SUBSTANCE: mixture of chitosonium glutamate obtained from high molecular chitosan with molecular mass 100-500 kD, and restrictedly depolymerised low polymeric chitosan with molecular mass 0.5-20 kD, is added to the inactivated flue vaccine as an adjuvant; the molecules of chitosan and chitosonium glutamate have free aldehydic groups at one of their free end, and have the deacetylation degree within 60-90%; the concentration of high molecular chitosan is brought to 0.5%, and low polymeric chitosan - to 0.05%, and the parenteral vaccination is performed.

EFFECT: method increases the immunogenicity of inactivated flue accine.

4 dwg, 4 ex

FIELD: medicine, veterinary.

SUBSTANCE: vaccine contains active substance and food additive. Active substance consists of efficient amount of avirulent antigenic material from "Novosibirsky" bird flue strain Influenzae virus avicum, Orthomyxoviridae family, serotype A, subtype H5N1, collection of FGU "VGNKI" VNIISZH N. 125 - deposition "Novosibirsky".

EFFECT: vaccine provides the efficient bird protection against the epizootic virus and prevents the environmental propagation of infectious agent from immunized bird.

6 cl, 12 tbl, 7 ex

FIELD: medicine, pharmacology.

SUBSTANCE: mentioned medicinal preparations (versions) include immunogenic protein or nucleic acid, coding the corresponding immunogenic protein (the sequences are presented in sequences list). Medicinal preparations, based on immunogenic protein or protein coding nucleic acid, are used in immunogenic formulation for humoral immunity induction, applied as pharmaceutical means. The invention also covers method of infectivity neutralisation of mentioned bacteria and method of patient immunisation against Chlamydia trachomatis infection by applying immunogenic compositions.

EFFECT: applying antigens and immunogens, identified by the invention, provides producing of more effective vaccines, diagnostic and treatment preparations.

7 cl, 162 dwg, 2 tbl, 44 ex

FIELD: medicine.

SUBSTANCE: invention refers to anti-infective materials (agents) and methods of production thereof, characterised by biocompatibility with animal or human body liquids and tissues. Invention concerns method of production of anti-infective agent, production of anti-infective agent implying modification of inorganic mineral with silico- and alumooxycompounds, namely, Na-shaped bentonite, inorganic metal salts in polar solvent, followed by bentonite keeping in salt solution, isolation of promodified bentonite from solution and drying at temperature not higher 100°C. Before being modified bentonite is enriched with Na+ ions through processing with 3-10% aqueous solution of chloride sodium with following washing and filtering of produced semiproduct thereafter modified by 10-20% inorganic metal salts solution represented with silver nitrate or copper sulphate. Modified bentonite is kept in specified salt solutions within 12-24 hour. Then promodified bentonite is cleared from sodium salts through washing and filtration. After being dried made agent is milled to particle dispersion 20-150 nm. Thus inorganic mineral are processed with specified solutions in ratio, weight fraction bentonite:solution, as 1: (10-40).

EFFECT: production of agent with higher anti-infective efficiency using simple and cheap methods and materials.

7 cl, 6 ex

FIELD: medicine; pharmacology.

SUBSTANCE: invention refers to new nucleoside analogues characterised as inhibitors of purinephosphoribosiletransferase, purinenucleosidephosphorylase, 5 '-methylthioadenosinephosphorylase, 5 '-methylthioadenosinenucleosidase and/or nucleosidehydrolase and can be applied for treatment of malignant neoplasm, bacterial infections, protozoal infections, T- cell mediated diseases. In formula V is selected of CH2 and NH, and W is selected of NR1 and NR2; X is selected of CH2 and CHOH in R or S - configurations; Y is selected of hydrogen and hydroxy; Z is selected of hydrogen, halogen and hydroxy, SQ and Q where Q is optionally substituted with halogen C1-C6alkyl or benzyl group; R1 is radical of formula ; R2 is radical of formula ; A is selected of N, CH; B is selected of OH, NH2 and halogen; D is selected of NH2 and hydrogen; E means N; G is selected of CH2 or G is absent. Invention also refers to based pharmaceutical composition, methods of disease or condition treatment requiring inhibition of purinephosphoribosiletransferase, purinenucleosidephosphorylase, 5 '-methylthioadenosinephosphorylase, 5 '-methylthioadenosinenucleosidase and/or nucleosidehydrolase, and to application of invention compounds for medical product preparation.

EFFECT: products are characterised with increased efficiency.

30 cl, 6 tbl, 19 dwg, 51 ex

FIELD: medicine.

SUBSTANCE: group of inventions concerns veterinary microbiology and immunology. Method of vaccine production includes cultivation of strain B.necrophomm "0-1" "ВИЭВ" on nutrient medium, inactivation with formalin, biomass isolation from culture fluid by centrifugation, centrifugate isolation and exotoxin ultrafiltration concentration to protein content 5.5-6.0 mg/ml (by Lowry). Exotoxin B.necrophomm of molecular weight 18000-20000 D is produced. It is added with formalin solution to end concentration 0.4% and inactivated within 15 days. Formalin inactivated exotoxin is mixed with oily adjuvant at the following component proportions (mass%): formalin inactivated exotoxin B.necrophorum - 60.0-67.0, oily adjuvant - 33.0-40.0. Then produced suspension is homogenised and end vaccine is packed up. A vaccine is injected intradermally in single or twice dosage 0.2-0.4 ml. Invention enables to simplify method of production of highly effective vaccine, to produce vaccine of reactogenity, as well as to lower vaccination dose and increase convenience of introduction.

EFFECT: development of effective method of necrobacteriosis vaccination in veterinary science.

10 cl, 4 ex

FIELD: medicine.

SUBSTANCE: invention refers to methods and compositions for treatment and/or prevention maintenance of infectious condition in liquid-containing organ or organ with natural external aperture. Composition includes antibacterial agent, and also, probably, other active agents, amphipathic oil which is soluble in water and insoluble in ethanol, microcrystalline wax and pharmaceutically acceptable nonaqueous carrier. The method of treatment is realised by introduction of a pharmaceutical composition in body through natural external aperture.

EFFECT: regarding composition, invention provides stability of the active agent against oxidising decomposition, low interfacial tension of composition, easy solubility in liquids, and short time of excretion; invention provides effective treatment and reduction of by-effects and toxicity.

61 cl, 12 ex

FIELD: medicine.

SUBSTANCE: can be used as method of treatment of osteomyelitis and septic arthritis, caused by bacterial agents, including with plural medicinal resistance. For this purpose mammal is introduced with pharmacologically effective amount of tigecycline separately or in combination with ryphampycine antibiotic. This agent is also offered also for manufacturing of medical products for treatment of given diseases.

EFFECT: invention provides increased efficiency of bone, marrow and joints infection treatment to preferential tigecycline distribution through bone tissue, marrow and synovial fluid.

48 cl, 5 dwg, 11 tbl, 3 ex

FIELD: chemistry.

SUBSTANCE: invention relates to the N" -substituted 9a-N-(N'- carbamoyl-γ-aminopropyl) and 9a-M-(M'-thiocarbamoyl-γ-aminopropyl)) derivatives of 9-desoxo-9-dihydro-9a-ase-9a-homoerythromycine A or 5-O-desosaminyl-9-desoxo-9-dihydro-9a-ase-9a-homoerythronolide A, novel semisynthetic macrolide antibiotics from the group of azalides with the common formula 1, where R means H or cladinosyl fragment, R means H, R means the isopropyl group, 1-naphtyl, 2-naphtyl, benzene, 2-(trifluoromethyl)phenyl, 3-phenylpropyl, (3-phenylethyl, ehtoxicarbonylmethyl, 1-(1-naphtyl)ethyl, 3,4,5-trimethoxiphenyl and 2,4-dichlorphenyl, and X means O or S, or to their pharmaceutically compliant additive salts with the inorganic or organic acids and to the method of their production. The invention also relates to the pharmaceutical composition with the antibacterial activity and to the application of the compounds with the common formula 1 or their pharmaceutically compliant salts, in producing the said composition.

EFFECT: antibacterial activity.

16 cl, 26 ex, 1 tbl

FIELD: medicine; pharmacology.

SUBSTANCE: ointment includes vegetable raw materials: sprouts and buds of poplar, aspen or willow, swallowwort (herb and blossom), common wormwood herb, yellow melilot (herb and blossom), costmary (herb and blossom), silver fir (sprouts), Siberian cedar (sprouts), pine resin and fat base, vegetable raw materials ratio is 1:10. As fat base composition contains unsalted pork fat.

EFFECT: extended range of preventive and therapeutic vegetable-based medicinal agents providing effective treatment of inflammatory diseases.

2 cl, 3 ex

FIELD: medicine; veterinary science.

SUBSTANCE: medicinal agent is spirit tincture made on basis of mixed Scotch pine buds, purple Echinacea herb and blossom clusters, horseheal rhizomes and roots, taken in proportion 2:1:1 with 70% concentrated ethyl alcohol 70% at raw and extractant ratio 1:5. Spirit tincture is exposed within 7 days in dark place at temperature 18-20°C. Calves ate treated within three courses every 48 hours with medicinal agent as 3.5-5.0% aqueous solution taken 2.0-3.0 ml/m3 during 45-60 minutes.

EFFECT: provided immunocorrection of key immunity disorders and increased natural resistivity of calves.

2 cl, 4 tbl, 6 ex

FIELD: medicine; pharmacology.

SUBSTANCE: invention is medicinal agent containing rybafutin, copolymer of lactic and glycolic acids (PLGA 50/50), D-mannitol, polysorbate 80 and dimethyl sulfoxide, as well as related method of production implying that mixed rybafutin, PLGA 50/50, D- mannitol, polysorbate 80 are heated at 50÷60°C and stirred until solid phase is completely dissolved, then cooled to room temperature, added with water at ratio 1 : 20 to 1 : 10 followed with nanoparticles˜200÷400 nm suspension produced as a result.

EFFECT: reduced toxicity of active substance accompanying maintained antimicrobic action and simplified production of end formulation.

3 cl, 4 ex, 4 tbl, 1 dwg

FIELD: dietology.

SUBSTANCE: the present innovation deals with the intake of a product containing chemically modified starch that provides the digestion of under 25% starch per 20 min, 30-70% per 120 min and above 60% per 240 min being measured by English's technique. The intake of such products provides a consumer with glucose for prolonged period of time and at more constant level.

EFFECT: higher efficiency of control.

30 cl, 2 dwg, 5 ex, 4 tbl

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