Secretory immunoglobulin a preparation possessing antiviral or antibacterial effect

FIELD: biotechnology, immunology.

SUBSTANCE: invention proposes preparation that comprises the immunoelectrophoretically pure secretory immunoglobulin A isolated from whey milk and/or colostrum of immunized ungulate animals and pharmaceutically acceptable vehicles. The base preparation (substance) comprises 6-12% of secretory immunoglobulin A at pH 4-8, an anti-complementary activity at least 10 mg of protein, not activating 2 CH50, protects in >70% against corresponding infections (in infection macroorganism in doses ≥10 ID50), shows areactogenic property in intravenous administration, can comprise stabilizing additives in the total concentration 4%, not above. The preparation possesses high purity, low anti-complementary activity, stable in storage, useful for oral, parenteral and topical using and possesses therapeutic activity with respect to microorganisms and viruses against which humans and animals immunization have been carried out. Invention can be used in treatment and prophylaxis of immunodeficiency states, bacterial and viral infections in humans and animals.

EFFECT: valuable medicinal and veterinary properties of preparation.

9 cl, 1 tbl, 10 ex

 

The invention relates to biotechnology and immunology, and can be used to obtain specific secretory immunoglobulin a medicine designed for treatment and prevention of immune disorders, bacterial and viral infections of humans and animals.

Preparations of immunoglobulins is a highly effective means of treatment and prevention of various infectious, viral and immunodeficiency diseases, but to date have been developed and used preparations of immunoglobulins from human serum.

Modern commercial preparations of serum immunoglobulin contains mainly IgG-immunoglobulin (85-93%) and only 1-3% of the immunoglobulins of other classes.

Currently, several countries are available and have been successfully used in the clinic preparations enriched IgM and IgA as for intramuscular and intravenous administration, however, despite extensive medical and veterinary use of drugs immunoglobulins as homologous and heterologous, there are currently no drugs secretory immunoglobulins intended for therapeutic and prophylactic use, allowing for a wide variety of routes of administration of such a drug.

Secretory immunoglobulin a and M are classes of immunoglobulins, ensuring the mi the body's defense against infectious agents directly at the gate of their penetration - in the skin and mucous membranes [1]. In terms of infectious process a major role in protecting the body plays locally synthesized antigen-specific secretory immunoglobulin. Antigen-specific secretory immunoglobulin a can agglutinate microbial body, to bind toxins [2], and to regulate the functional activity of immune cells in inflammation [3], preventing generalization of the infectious process and the development of systemic immune response. Specific secretory immunoglobulin a is produced as the bacteria [4]and viral [5, 6, 7] antigens. Moreover, in models of viral infections shown that secretory immunoglobulin a is not only able to interact with the virions on the surface of the mucous membranes, but also penetrate into the infected cells, there blocking virus replication.

The difference of secretory immunoglobulins from serum immunoglobulins is the ability to penetrate intact mucous membranes and skin and is long to remain in secreted fluids.

Despite the fact that the drug-specific polyclonal human secretory immunoglobulin a would be of great clinical interest, obtaining it from human colostrum or milk would provide immunization be the b women or use as donors pregnant women undergoing within 1 month before birth or during lactation certain infectious disease. The complexity of the selection of donors such severely limit the development and practical use of such drugs. Therefore, of particular interest for industrial production are drugs heterologous immunoglobulin A.

Well-known medication for the treatment of multiple sclerosis, in which as an active ingredient use colostrum from pregnant cattle immunized with the measles virus and/or virus Onderstepoort [8]. The disadvantage of this drug is that it is derived from inaccessible sources, and that it is intended solely for internal (oral) applications. On the other hand, it is known that secretory immunoglobulins practically does not penetrate into the circulation through the intact mucosa of the gastrointestinal tract of man, therefore, to achieve therapeutic effect it is necessary to use very high doses of the drug.

The most closest to the claimed drug prototype is immunological drug for the treatment of bacterial infections, which includes a purified complex immune proteins, including immunoglobulin a, obtained from milk or colostrum from immunosera the different cows for oral use [9]. Well-known medication contains 95% of immunoglobulins and up to 5% impurities nimmanahaeminda nature, while immunoglobulins represented mainly IgG (82.7 per cent) and also contain IgA (10.4%) and IgM (6,9%).

The disadvantages of this drug is its lack of cleanliness, a considerable admixture of classes of immunoglobulins M and G, which limits the scope only to the treatment of intestinal infections and the fact that it is intended only for oral administration.

An object of the invention is the creation of a high-purity preparation of secretory immunoglobulin, with a wide range of therapeutic efficacy, suitable for oral, parenteral and topical use.

The problem is solved in that the proposed product containing secretory immunoglobulin A (S-IgA) with a purity of not less than 99,0%, isolated from milk and/or colostrum of immunized ungulates and pharmaceutically suitable excipients.

Basic drug (substance) contains 6-12% of secretory immunoglobulin a has a pH of 4-8, anticomplementary activity of not less than 10 mg of protein, not activating 2 CH50protects >70% of the respective infections when the infecting microorganism in doses >10 EID50is reactogenic when administered intravenously, may contain a stabilizing additive in common is concentratie not more than 4%.

On the basis of the substance can produce different dosage forms:

1. Sterile lyophilized preparation of secretory immunoglobulin a for parenteral, oral and topical use

2. Tablets and/or Calculadora form of the drug for oral administration, containing from 0.2 to 0.5 g of active start and auxiliary substances in quantities permitted for use in the composition of medicines

3. Ointment form or gel form, containing from 1% to 25% of the current start and auxiliary substances in quantities permitted for use in the composition of medicines.

4. Rectal and vaginal suppositories containing from 0.2 to 1.5 g of active start and auxiliary substances in quantities permitted for use in the composition of medicines

The method of obtaining the proposed drug is as follows.

Pregnant cows are immunized 2 times live or synthetic vaccine. 1st immunization is held 40 days before calving, 2nd immunization - 10 days before calving. 1st immunization (premirovanii) is used to create a booster of the immune system, is carried out in a dose of 0.1 EID50when this half is administered intramuscularly, half in the tissue of the udder. 2nd immunization (resolution) is carried out at a dose of 10 EID50intramuscularly.

Molo is Ivo and the milk is harvested 2-3 days after birth. Fat is removed by centrifugation, the aqueous phase is acidified with 1 n acetic acid to a pH of 4.6 to precipitate the casein. Then hold clearing the drug from ballast substances and aggregated immunoglobulin. For this immunoglobulin is precipitated by ammonium sulfate at 40% saturation and dissolved in distilled water and cialiswhat first against distilled water for disaggregation IgG, and then against the starting buffer of 0.02 M Tris-HCl, pH 7.4 and at the end of dialysis chromatographic on the column with DEAE-cellulose. Elution is conducted stepwise gradient of Tris-HCl-buffer pH 7.4: 0.02 M, 0.05 M, 0.75 M, 0.1 M, 0.125 M, 0.15 M, each stage contains 200 ml of the corresponding buffer. IgA eluted 0,075 M and 0.1 M buffers. This fraction contains IgA and S-IgA, which vary widely in molecular weight. This allows you to divide them by the method of gel filtration. Use mainly the Sephadex G-200. Before chromatography, the solution is concentrated by ultrafiltration to a volume of 40 ml and applied to a column of 300 ml, 2×60 see First eluted S-IgA. The yield of the target product is 6-10 g of 1 l of raw materials.

Drug test for specific activity by establishing a titer of specific antibodies in reactions of indirect haemagglutination reaction of inhibition of hemolysis, enzyme immunoassay, radioimmunoassay, in neutralization reactions n the cell cultures and animal models and other methods.

The drug immunoelectrophoretic homogeneous.

The final form of the drug may be either liquid or lyophilized. To obtain a liquid form, the protein concentration was adjusted to 6 to 12%, set the pH value of 4.0 to 8.0, enter stabilizers selected from the group glycine, nicotinamide, glucose, then the immunoglobulin is subjected to sterilizing filtration and poured into bottles for the dry form of the drug after filling the bottles lyophilizer.

Determining significant difference between the claimed product from the preparation of the prototype is that as immunoglobulin use immunoelectrophoretic net secretory immunoglobulin a (S-IgA) and pharmaceutically suitable excipients, which provides an opportunity not only for domestic but also for intravenous administration of the drug at the expense of a higher quality. The main quality indicators of intravenous immunoglobulin (IV IG) is low anticomplementary activity (AKA), the absence of aggregates, low dimers and fragments, the higher content of monomers and stability of these indicators during storage.

The drug has the following quality indicators:

Specific activity (on the neutralization in vitro and in vivo): protects >70% of the respective infections if the infection microorganism is and in doses ≥ 10 EID50.

Anticomplementary activity: not less than 10 mg of protein, not activating 2 CH50.

Purity (immunoelectrophoretic purity or content of immunoglobulins of other classes) is 0-1%.

Number of immunologically inactive impurities: 0-1%.

The content of fragments: not more than 10% at the end of the retention period.

The storage stability: 2 years when stored in a dark place at a temperature of +4+6°C.

The invention is illustrated by the following examples of specific obtain different forms of the proposed drug.

Example 1. Getting dry bullish smallpox secretory immunoglobulin A.

Pregnant cows were immunized 2 times live spooktinu. 1st immunization was carried out for 40 days before calving, 2nd immunization - 10 days before calving. 1st immunization (premirovanii) was used to create a booster of the immune system, was carried out in a dose of 0.1 EID50when this half was injected intramuscularly, half - cloth - udder. 2nd immunization (resolution) was performed at a dose of 10 EID50intramuscularly. Colostrum and milk began to gather in 2-3 days after birth. The fat was removed by centrifugation at 20000 rpm./minutes within 2 hours the Aqueous phase was acidified using 1 n acetic acid to a pH of 4.6 to precipitate the casein.

Immunoglobulin precipitated with ammonium sulfate at 40% saturation was dissolved in di is fillerbunny water and were dialyzed first against distilled water at a temperature of 2-4° C for 12-15 h to disaggregate IgG, and then against the starting buffer of 0.02 M Tris-HCl, pH 7.4 and at the end of dialysis was chromatographically on the column with DEAE-cellulose, with a volume of 200 ml Elution was performed stepwise gradient of Tris-HCl-buffer pH 7.4: 0.02 M, 0.05 M, 0.75 M, 0.1 M, 0.125 M, 0.15 M, each stage contained 200 ml of the corresponding buffer. IgA was elyuirovaniya of 0.075 M and 0.1 M buffers. This fraction contained IgA and S-IgA, which vary widely in molecular weight. It is possible to divide them by the method of gel filtration. Used Sephadex G-200. Before chromatography, the solution was concentrated by ultrafiltration to a volume of 40 ml and was applied on a column of 300 ml, 2×60 see First elyuirovaniya S-IgA. The yield of the target product was 7 g per 1 l of raw materials. The drug immunoelectrophoretic homogeneous. The protein concentration was brought to 12% with diafiltration against 0.1 M phosphate buffer, pH set to a value of 4.5±0,4, introduced stabilizer glycine in the amount of 25 g/l, the resulting solution of immunoglobulin was subjected to sterilizing filtration. The result has been a preparation containing, in wt.%:

S-IgAto 12.0
Glycine- 2,5
phosphate buffer- the rest.

The resulting preparation was poured into the Lacona and liofilizirovanny.

Example 2. Getting liquid horse smallpox secretory immunoglobulin for intravenous injection.

Immunization, remove from milk and colostrum fat and casein was carried out similarly to example 1.

Immunoglobulin precipitated with ammonium sulfate at 40% saturation was dissolved in distilled water and were dialyzed first against distilled water at a temperature of 2-4°C for 12-15 h to disaggregate IgG, and then against the starting buffer 0.01 M phosphate buffer, pH 7.0, and at the end of dialysis was chromatographically on the column with DEAE-cellulose, with a volume of 200 ml Elution was performed stepwise gradient of phosphate buffer: 0.01 M, pH 7.4; 0.02 M, pH 7,2; 0,06 M, pH 7.0; 0.1 M, pH 6.0; 0.2 M, pH 6.0 each stage contained 200 ml of the corresponding buffer. IgA was suirable 0,06 M buffer with pH 7.0. Gel filtration was performed on Sephadex G200. Before chromatography, the solution was concentrated by ultrafiltration to a volume of 40 ml and was applied on a column of 300 ml, 2×60 see First eluted S-IgA. The yield of the target product was 9 g per 1 l of colostrum. The drug immunoelectrophoretic homogeneous. The protein concentration was brought to 6% with a 0.9%solution of sodium chloride, the pH was set to 7.0±0,4, introduced stabilizers glycine to 22.5 g/l nicotinamide 7.5 g/l, the resulting solution was subjected to sterilizing filtration, and the difference is Ali vials. Received liquid medication, primarily for intravenous administration, with anticomplementary activity 15 mg protein does not activate 2 CH50containing the following components in wt.%:

S-IgA- 6,0
Glycine- 2,25
Nicotinamide- 0,75
Saline- rest

Example 3.

The preparation of secretory immunoglobulin a was obtained analogously to example 1, except that the concentration of protein was brought to 8.0%, and the stabilizer used nicotinamide in an amount of 10 g/L. as a result received a preparation containing, in wt%:

S-IgA,- 8,0
Nicotinamide- 1,0
phosphate buffer- the rest.

The resulting preparation was poured into the bottles and liofilizirovanny.

Example 4.

The preparation of secretory immunoglobulin a was obtained analogously to example 1, except that the concentration of protein was brought to 10%, and as stabilizers used glucose (20 g/l glycine in the amount of 20 g/L. as a result received a preparation containing, in wt%:

S-IgA- 10,0
Glycine- 2,0
Glucose- 2,0
phosphate buffer- the rest.

The resulting preparation was poured into the bottles and liofilizirovanny.

Example 5. Antiherpes virus effect of the gel.

Immunization, isolation and purification of substances secretory IgA was carried out similarly to example 1. After adding the necessary components received Antiherpes virus effect of the gel containing in wt.%:

The methylcellulose- 3%
Dried antiherpetic S-IgA- 5%
glycerin- 20%
methylparaben- 0,1%
distilled waterup to 100%

The gel is prepared as follows. Methylcellulose pour half from the calculated volume of water at a temperature of 70°With leave to stand for swelling to cool to room temperature. Prepare a 15% aqueous solution antiherpetic secretory immunoglobulin a and add the calculated amount to the swollen methylcellulose. The volume was adjusted with water containing a definite number of methylparaben up to 80% on the final and stirred by a mechanical stirrer at 3000 rpm, to policereports homogeneous mass, then under stirring injected glycerin. Prescription weight Packed in tubes.

Example 6.

For the manufacture of the drug in the form of tablets taken (per tablet weighing 0.5 g): lyophilized S-IgA - 0.2 g, components for tabletting of 0.3, In the rotary type mixer load the sample is pre-crushed and occasioanly components, carefully mix the mixture for 15 min and produce tableting at a rotary tablet machine. The finished tablets are packaged in bottles and/or cans.

Example 7.

For the manufacture of the drug in the form of capsules taken (one capsule), g: lyophilized S-IgA, - 0.3, and components for encapsulation of 0.3. In the rotary type mixer load the sample is pre-crushed and occasioanly components, carefully mix the mixture for 15 min and produce encapsulation on a special installation. The finished capsules are packaged in bottles and/or cans.

Example 8.

For the manufacture of the drug in the form of ointment take, in wt.%:

Lyophilized S-IgA - 10,0; anhydrous lanolin - 25,0; vaseline medical - 50,0; preservative - 0,5; water - the rest.

The ointment is prepared as follows.

In a reactor with a stirrer at room temperature load anhydrous lanolin and calculated the amount of water treated. A lot emuleret 20-30 minutes. In the reactor the resulting aqueous Lano is another load vaseline medical and stirred for 20-30 minutes. Then add preservative (methyl ether of peroxybenzoyl acid) and S-IgA. Mixing of all components is carried out for 20-30 minutes, after which the product Packed in the container.

Example 9.

For the manufacture of the drug in the form of suppositories weighing 2.4 g take, in g (one suppository): lyophilized S-IgA - 0.2 g protector - 0.36 g, target additive (emulsifier, stabilizer, solvent, confectionery fat, paraffin) - 1,84 was charged To the reactor prescription number of the target additives and heated to their melting, the mixture is then emuleret at 70°C for 40 min, cooled, the cooled mixture is injected components protector and S-IgA, mix until smooth and Packed

Example 10.

Immunization of pregnant cows spent monovalent adsorbed diphtheria toxoid twice: the first time in 2 months. prior to calving dose of 0.25 ml subcutaneously in the second - 10-14 days before calving at a dose of 1.0 ml With 2-s introduction was carried out as follows: 0.75 ml subcutaneously and 0.25 ml in the tissue of the udder. Milk and colostrum started to collect 2-3 days after calving.

The preparation of secretory immunoglobulin a was obtained analogously to example 1. The liquid intravenous drugs had antitoxic activity of 40 IU/ml

Antitoxic activity of the obtained preparation was compared with protiva verinym human immunoglobulin in patients with a toxic form of diphtheria.

Experimental group patients and the comparison group were matched for age, severity and localization of diphtheria, duration of hospitalization and the beginning of the specific therapy, the use of pathogenetic and symptomatic means. Both drugs were administered in the course dose of 250 IU/kg of body weight, intravenously, twice with an interval of 12 hours.

The results of a comparative study of the activity of diphtheria bullish secretory immunoglobulin and human diphtheria serum immunoglobulin presented in the table.

SymptomBullish antilittering S-IgA (n=20)Diphtheria immunoglobulin human (Tomsk research Institute of vaccines and sera) (n=20)
IIIIII
Fever202,38±1,22202,28±1,11
Lethargy203,66±1,74203,54±2,01
Loss of appetite202,88±1,91202,74±1,72
Pale skin202,61±2,00 202,58±2,11
Sore throat202,01±0,53203,13±1,6b
Enlarged lymph nodes20of 2.51±0,42202,74±0,88
The density of lymph nodes202,92±1,18203,79±1,B8
Tenderness of lymph nodes201,39±0,66*202,55±0,75
Edema of the oropharynx202,22±0,94*203,62±1,43
Common RAID202,14±0,44*203,98±1,33
Peloponnesiaca plaque201,02±0,34*201,8b±0,32
Bleeding of the mucous membrane after removing plaque70,38±0,11*201,33±0,21
Edema of the subcutaneous tissue202,69±1,57202,77±1,28
Nasal breathing201,58±0,42*172,66±0,65
* - significant differences, p<0,05

The table shows that having comparable total antioxidant activity, secretory immunoglobulin superior to whey local manifestations of diphtheria infection, in particular accelerated the elimination of the inflammatory phenomena in the oropharynx and the nasopharynx, facilitated the discharge of fibrinous film, reduces the severity of the reaction regional lymph nodes. Prophylactic activity of both drugs in relation to complications in the internal organs was close, but the preparation of diphtheria bullish secretory immunoglobulin has a greater therapeutic activity in localized forms of diphtheria and comparable with serum immunoglobulin in toxic forms of this disease.

Thus, the proposed immunoglobulin preparation with therapeutic activity (action) in respect of those bacteria or viruses, against which were immunized animals. This is evidenced by the term "specific", i.e. directed against the infection, against which were immunized and not against all infections. The inventive preparation of S-IgA, unlike all other drugs colostrum can be applied parenterally, and not only inside, so the drug can be used, including, when terinfeksi, while traditionally impose specific serum immunoglobulin. The inventive preparation is not universal in respect of any infection bacterial or viral nature, and is, like all immunoglobulin preparations, specific S-IgA, which will be used only for those infections, against which were immunized animals.

Developed specific drug secretory immunoglobulin a has a high activity of antibodies against bacterial or viral infections, low anticomplementary activity, stable during storage, suitable for wide use in medical practice for the treatment and prevention of immune disorders, bacterial or viral infections of humans and animals.

SOURCES of INFORMATION

1. Kerr MA. The structure and function of human IgA. Biochem.J.,1990, 271, 1553-1559.

2. Jarvis GA, GriffisJM. Human IgAl blockade of IgG-initiated lysis of Neisseria meningitides is a function of the antigen-binding fragment binding to polessaccharide capsule J.ImmunoL, 1991, 147, 1962-1967.

3. Kaetzel CS, Robinson JK, Chintalacharuvu KR et al. The polymeric immunoglobulin receptor (secretory component) mediates transport of immune complexe across epithelial cells: a local defense function for IgA. Proc. Natl. Acad. Sci. USA, 1991, 88, 8796-8800.

4. Glass M, Svennerholm A-M, Stoll BJ, et al. Protection against cholera in breast-fed children by antibodies in breast milk. N. Engl. J. Med., 1983, 108, 1389-1392.

5. Mazanec MB, Nedrud JG, Lamm ME. Immunoglobulin A monoclonal antibodies protect against Sendai Virus. J.Virol., 1987, 61, 2624-2626.

6. Turner RB, Kelsey DK. Passive immunization for pevention of rotavirus illness in healthy infants. J. Infect Dis., 1987, 156, 158-166.

7. Renegar KB, Small PA. Immunoglobulin A mediation of murine nasal anti-influenza virus immunity. J.Virol., 1991, 65, 2146-2146.

8. Accepted Japan's bid No. 4-43888, class. AND C 39/42, publ. 10.03.82.

9. U.S. patent No. 5017372, class a 61 K 39/395, publ. 21.05.91.

1. Immunoglobulin preparation with specific antiviral or antibacterial action, containing immune proteins derived from milk and/or colostrum of immunized hoofed animals, characterized in that as the immune protein it contains secretory immunoglobulin a with immunoelectrophoretic purity not less than 99,0%, a stabilizer and a pharmaceutically suitable excipient in the following content, wt.%:

Secretory immunoglobulin a6,0-12,0
Stabilizer1,0-4,0
FillerRest

2. Immunoglobulin preparation according to claim 1, characterized in that it contains a stabilizer selected from the group: glycine, glucose, nicotinamide.

3. Immunoglobulin preparation according to claim 1, characterized in that it contains glycine in the amount of 2.0 to 3.0 wt.%, glucose in the amount of 2.0 to 3.0 wt.%, nicotinamide in the number of 0-1,0 wt.%.

4. Immunoglobulin preparation according to claim 1, characterized in that the filler it contains phosphate is a buffer or saline solution.

5. Immunoglobulin preparation according to claim 1, characterized in that it has a pH of 4.0 to 8.0.

6. Immunoglobulin preparation according to claim 1, characterized in that it is provided in lyophilized form for parenteral or topical application.

7. Immunoglobulin preparation according to claim 6, characterized in that it is presented in tablet or capsule form for oral administration with the content of secretory immunoglobulin a in the amount of 0,2-0,5 g

8. Immunoglobulin preparation according to claim 6, characterized in that it presents in ointment or gel form, containing 5-10 wt.% secretory immunoglobulin a for local use.

9. Immunoglobulin preparation according to claim 6, characterized in that it presents in the form of rectal and vaginal suppositories with the content of secretory immunoglobulin a in the amount of 0,2-0,5 g



 

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10 cl, 4 sch, 4 tbl, 9 ex

FIELD: organic chemistry, biochemistry, pharmacy.

SUBSTANCE: invention relates to new heterocyclylsulfonyl alkylcarboxylic acids and their derivatives of the general formula (1): or their pharmaceutically acceptable salts, N-oxides or hydrates possessing the inhibitory effect on kinase activity and to the focused library for search of active leader-compounds comprising at least abovementioned compound. In the general formula 91) W represents optionally substituted heterocyclic radical, among them: pyrrole-3-yl, thiophene-2-yl, isooxazole-4-yl, pyrazole-4-yl, imidazole-4-yl, pyridine-3-yl, 1H-2,4-dioxopyrimidine-5-yl, 2,3-dihydro-1H-indole-5-yl, 2,3-dihydro-1H-indole-7-yl, 1,3-dihydro-2-oxoindole-5-yl, 2,3-dioxo-1H-indole-5-yl, 2-oxo-3H-benzoxazole-6-yl, benzothiazole-6-yl, 1H-benzimidazole-5-yl, benzo[1,2,5]oxadiazole-4-yl, benzo[1,2,5]thiadiazole-4-yl, 1,2,3,4-tetrahydroquinoline-6-yl, 3,4-dihydro-2-oxo-1H-quinoline-6-yl, quinoline-8-yl, 1,4-dihydro-2,3-dioxoquinoxaline-6-yl, 3-oxo-4H-benzo[1,4]oxazine-7-yl, 3-oxo-4H-benzo[1,4]thiazine-7-yl, 2,4-dioxo-1H-quinazoline-6-yl, 2,4-dioxo-1,5-dihydrobenzo[b][1,4]diazepine-7-yl or 2,5-dioxo-3,4-dihydrobenzo[b][1,4]diazepine-7-yl; Y represents optionally substituted methylene group; R1 represents chlorine atom, optionally substituted hydroxyl group, optionally substituted amino-group, optionally substituted azaheterocyclyl; n = 1, 2 or 3; or Yn represents carbon atom of optionally substituted (C3-C7)-cycloalkyl or optionally substituted (C4-C7)-heterocyclyl. Also, invention relates to a pharmaceutical composition in form of tablets, capsules or injections placed into pharmaceutically acceptable package.

EFFECT: valuable properties of compounds.

5 cl, 3 sch, 5 tbl, 6 ex

FIELD: organic chemistry, chemical technology, antibiotics.

SUBSTANCE: derivative of 9-deoxo-9a-aza-9a-homoerythromycin A of the formula (3) wherein R4 represents hydroxyl protecting group is prepared by protection of 2'-hydroxy-group of compound of the formula (5) to form compound of the formula (4)

and by oxidation of C-4''-hydroxy-group of compound of the formula (4) that is carried out by addition of dimethylsulfoxide to solution containing compound of the formula (4) and a solvent followed by cooling the mixture up to about -70°C, activation of dimethylsulfoxide in situ and defoaming the reaction mixture. Compound of the formula (4) is converted to the oxidation stage directly without its isolation. Also, invention proposes additive salt of trifluoroacetic acid of compound of the formula (3) and a method for its preparing by treatment of compound of the formula (3) with trifluoroacetic acid. Invention provides increasing yield and improving purity of the end product.

EFFECT: improved preparing method.

11 cl, 6 ex

FIELD: medicine.

SUBSTANCE: the present innovation deals with infecting macrophages and cells of macrophagous cell lines with pathogenic microorganisms, their treatment with medicinal remedies followed by gamma-irradiation, immunization of disease-resistant and disease-sensitive animal lines and infecting vaccinated animals with alive pathogenic microorganisms that leads to improved immunity to pathogenic microorganisms.

EFFECT: higher efficiency.

4 cl, 1 dwg, 3 ex

FIELD: medicine, gynecology.

SUBSTANCE: one should perform laparoscopy, drain purulent foci, remove destructive tissues and wash abdominal cavity with great amount of "Baliz-2" introduced into abdominal cavity by fractional portions per 1-1.5 l at exposure up to 3 min either once or up to three times. The method provides complex impact as antibacterial, antioxidant and immunocorrecting actions of the preparation in area of lesion and, thus, quick interruption of acute stage by preventing the development of adhesions and saving reproductive function in this category of patients.

EFFECT: higher efficiency of therapy.

2 ex

FIELD: biotechnology, microbiology, medicine, veterinary science.

SUBSTANCE: for preparing vaccine toxigenic strains of S. dysenteriae R-forms are grown, cells are subjected for lysis by treatment with chloroform, mixture is centrifuged and prepared supernatant is treated with saturated monobasic carboxylic acid or their derivatives and pH value is brought about to 3.0-5.0. Mixture if centrifuged and precipitate containing corpuscular antigens and shigellosis exotoxin are obtained. Precipitate is dissolved in buffer and pH value is brought about to 7.5-9.0. Then formalin is added in the amount 0.4-0.8% of the solution volume, or benzoic acid or benzoic acid salts are added in the amount 0.07-4.0% of the solution volume, or a mixture consisting of formalin and benzoic acid or benzoic acid salt solutions in the amount 0.10.3% and 0.03-2.5% of the solution volume, respectively. The solution is kept at temperature 30-60°C for 2 h - 60 days to provide the conversion of exotoxin to anatoxin and vaccine is prepared. Another variant of the claimed invention involves additional treatment with formalin or benzoic acid or benzoic acid salts to provide conversion of exotoxin to anatoxin, vaccine is prepared followed by its bagging and corking. For preparing immunoglobulin preparation animals are immunized with vaccine prepared by abovementioned methods followed by taking off blood, milk and/or colostrums, and/or blood, immunoglobulin fraction is prepared, sterilized, bagged and corked. This preparation is a component of the immunobiological preparation. The immunobiological preparation comprises the immunoglobulin preparation and at least one component taken among the following row: human and/or animal immunoglobulin preparations, lactoferrin, enzymes, inhibitors of proteolytic enzymes, human and/or animal normoflora preparations, yeast, vitamins, vitamin-like substances, human and/or animal proteins of acute phase, human and/or animal cytokines, higher plants components, lower plants components, components of natural origin products, apiculture products, enterosorbents, antibiotics, antibacterial chemopreparations, sulfanilamide drugs, antibacterial, anti-tuberculosis, antiviral preparations, antifungal antibiotics, synthetic antifungal preparations, stimulators of metabolic processes, antioxidants, mineral supplements, carbohydrates, lipids, replaceable and/or essential amino acids, organic acids, alkaloids, glycosides, taste supplements, aromatic supplements, base for suppositories, base for ointment formulations, technological additives for tableting, or their mixture. Invention provides preparing preparations eliciting antigenic activity with respect to broad species of pathogenic and opportunistic gram-negative microorganisms of intestine group and their exotoxins and therefore eliciting with prophylactic and curative effect with respect to diseases causing with these microorganisms.

EFFECT: improved preparing method, valuable properties of vaccine.

13 cl, 112 ex

Antibacterial agent // 2262346

FIELD: biotechnology, microbiology.

SUBSTANCE: invention proposes applying Flavobacterium odoratum culture as an antibacterial agent isolated from drinking mineral water "Ust-Kachkinskaya" from the hole 1/99. Microorganism Flavobacterium odoratum inhibits growth of Staphylococcus aureus, colon bacillus and yeast-like fungi Candida albicans that allows using microorganism Flavobacterium odoratum as an antibacterial agent. Invention can be used as an antibacterial agent.

EFFECT: valuable medicinal properties of agent.

2 ex

FIELD: biotechnology, medicine, antibiotics.

SUBSTANCE: invention proposes to the new compound amycomycin of the molecular formula C65H115NO18 (structural formula is given on the invention claim) that shows an antibacterial activity. Amycomycin, its pharmaceutically acceptable salts and derivatives in all their stereoisomeris and tautomeric forms can be obtained by culturing microorganism Amycolatopsis sp. ST 101170 (DSM 12216) under aerobic conditions on the nutrient medium containing the necessary nutrient components. The end product is isolated and purified and converted if necessary to its pharmacologically acceptable salt, ester, ether and other chemical derivatives and eliciting the same spectrum of antibacterial activity. Amycomycin is a component of the pharmaceutical composition eliciting an antibacterial activity. Amycomycin acts as an antibiotic. Invention provides inhibition of microorganisms with resistance to vancomycin and teicoplanin used in treatment of infections caused by Staphylococcus aureus.

EFFECT: improved preparing method, valuable medicinal properties of amycomycin.

7 cl, 2 tbl, 4 ex

FIELD: biochemistry, medicine.

SUBSTANCE: invention relates to two forms of peptide isolated from annelid worm (Arenicola marina) eliciting the broad spectrum of antibacterial effect. Indicated forms differ by a single amino acid residue at position 10 wherein at position 10 arenicin-1 has Val and arenicin-2 has Ile. Invention provides expanding assortment of antibacterial agents.

EFFECT: valuable medicinal properties of peptides.

1 tbl, 3 dwg, 4 ex

FIELD: medicine, gynecology, pharmacy.

SUBSTANCE: invention proposes using pimafucin (natamycin) as agent for treatment of bacterial vaginitis. Method for treatment of bacterial vaginitis involves intravaginal administration of pimafucin as 2% cream, 2 times per a day in morning and evening for 7-10 days and for first 3 days pimafucin-containing vaginal suppository in the dose 100 mg is administrated additionally in evening after administration of cream into vagina. Invention provides high effectiveness of treatment and clinical-etiological recovery in 92.3% of cases being without prescription of medicinal preparations. Method has no contraindications and can be used in all period of pregnancy and without adverse effects.

EFFECT: improved treatment method, enhanced effectiveness of agent and treatment.

2 cl, 1 ex

FIELD: medicine, narcology.

SUBSTANCE: invention involves using immunogenic monoclonal antibodies raised to opiates as an agent used in prophylaxis and treatment and treatment of opiomania. Invention provides expanding assortment of pathogenetic agents used in treatment and prophylaxis of opiomania and enhancing effectiveness in treatment. Invention can be used in prophylaxis and treatment of opiomania.

EFFECT: enhanced and valuable properties of agent.

2 cl, 1 tbl, 6 ex

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