Novel polycyclic xanthones and their using

FIELD: organic chemistry, microbiology, medicine, oncology.

SUBSTANCE: invention relates to novel class of antitumor compounds found in isolation from new marine microorganisms of genus Actinomadura sp., strain PO13-046, namely, to compound denoted as IB-00208: and relates to class of compounds of the formula (I) or (II) , wherein each substitute R1 is similar or different and can represent hydrogen atom, acyl, alkyl, alkenyl, aryl, benzyl, alkaline metal and/or monosaccharide, disaccharide, trisaccharide or their derivative; R2 and R3 can represent hydrogen atom or alkyl. These compounds show antitumor activity.

EFFECT: valuable medicinal properties of compounds.

8 cl, 3 tbl, 9 dwg, 11 ex

 

The present invention relates to polycyclic the xanthones and their anticancer use.

BACKGROUND of the INVENTION

Carminomycin (EP 00246091; EP 0054801; J. Antibiot., 1982, 35, 645-652; J. Am. Chem. Soc., 1986, 108, 6088-6089, J. Antibiot., 1987, 40, 301-308; J. Antibiot., 1994, 47, 342-348; synthetic derivatives J. Antibiot., 1986, 39, 1636-1639 and EP 0259496) and tetramycin (J. Antibiot., 1989, 42, 846-851; J. Antibiot., 1990, 43, 504-512; EP 0405151) belong to the family of naturally occurring antibiotics, each of which contains xantanovu structural unit included in a larger polycyclic structure.

These compounds had strong activity against aerobic and anaerobic bacteria and mycoplasmas, however, the antitumor activity was described only for carminomycin (EP 0246091).

Recently in the international application PCT/GB01/02148 authors of the invention has also been described antitumor activity of zithromycin.

For the treatment of many human cancers, there is still a need for new anticancer drugs. Accordingly, the present invention was the creation of new anticancer agents with polycyclic xantanovu structure.

Another objective of the present invention was the creation of pharmaceutical compositions for the in troduction the possible active compounds in need of treatment to patients.

Another objective of the invention has been directed to the production of polycyclic xanthone by the controlled aerobic fermentation using a biologically pure culture of the microorganism in a suitable nutrient medium, and to develop ways of selection and concentration of the fermentation broth, and final purification of the active compounds.

BRIEF description of the INVENTION

The present invention relates to a new class of active Kantorovich compounds, based on the opening of a new isolated from bacteria of the compounds IB-00208, useful as an antitumor drug of the formula:

Thus, the present invention relates to compounds of General formula:

or

where each substituent R1is the same or different and can represent hydrogen, acyl, alkyl, alkenyl, aryl, benzyl, alkali metal and/or sugar, and R2and R3can represent hydrogen, alkyl or together form an unsaturated bond.

The present invention also relates to methods of producing these compounds, including a method of obtaining compound IB-00208.

DETAILED description of the INVENTION

IB-00208 and related compounds obl is give antitumor activity against tumors in mammals, such as the human lung carcinoma, carcinoma human colon, melanoma of the person, and the like. Thus, the present invention includes a method of treating any mammal, suffering from a malignant tumor sensitive to it, which includes an introduction to the patient a therapeutically effective amount of the compounds or pharmaceutical compositions.

The present invention also refers to pharmaceutical preparations which comprise as active ingredient a compound IB-00208 or any derivation thereof, or their pharmaceutically acceptable salt, as well as to methods for their preparation. Used pharmaceutically acceptable carrier.

Pharmaceutical compositions typically derived from active compounds and include any solid forms (tablets, pills, capsules, granules, and the like) or liquid form (solutions, suspensions go emulsion) in combination with any carrier or other pharmacologically active compounds.

The correct dose of the pharmaceutical compositions of the compounds IB-00208 or its derivatives may be changed in accordance with a specific connection, preparative form, method of administration, and the localization of the owner and the type being treated tumors.

Other factors, such as age, body weight, sex, diet, time of administration, the speedy is here excretion, the patient's condition, combination of drugs, individual sensitivity and the severity of the disease, should be taken into account. The introduction can be effected continuously or periodically within the maximum tolerated dose.

Acyl group may be an aliphatic acyl, aromatic acyl or mixed aliphatic/aromatic acyl. For example, they may correspond to the formula RCO-, where R represents an alkyl group, alkenylphenol group, aryl group, arylalkyl group or alcylaryl group. Examples include benzoyl.

Alkyl groups typically contain from 1 to 18 carbon atoms. The alkyl groups preferably contain from 1 to about 12 carbon atoms, more preferably from 1 to about 8 carbon atoms, even more preferably from 1 to about 6 carbon atoms, and most preferably 1, 2, 3 or 4 carbon atoms. Especially preferred alkyl groups in the compounds of the present invention are methyl, ethyl and various, including ISO-propyl. Used herein, the term "alkyl", unless otherwise specified, refers to both cyclic and acyclic groups, although cyclic group must contain in the ring at least three carbon atoms. Alkyl groups can b the th with unbranched or branched chain.

Alkeneamine groups typically contain from 1 to 18 carbon atoms. Preferred alkeneamine group in the compounds of the present invention contain one or more unsaturated linkages and from 2 to about 12 carbon atoms, more preferably from 2 to about 8 carbon atoms, even more preferably from 2 to about 6 carbon atoms, and even more preferably 2, 3 or 4 carbon atoms. Used herein, the term "Alchemilla" refers to cyclic and acyclic groups, although unbranched or branched acyclic groups are in General preferred.

Aryl groups can be carbocyclic or heterocyclic and may contain one or more condensed rings. Carbocyclic aryl groups typically contain 6 or 10 carbon atoms in phenyl or naftilos groups. Heterocyclic aryl groups typically contain from 5 to 12 atoms, more often 4, 5, 6, 10, 11 or 12 atoms of which 1, 2, 3 or more are heteroatoms typically selected from oxygen, sulfur or nitrogen. Suitable heterocyclic aryl groups in the compounds of the present invention include coumarinyl, including 8-coumarinyl, chinoline, including 8-chinoline, pyridyl, pyrazinyl, pyrimidyl, furyl, pyrrolyl, thienyl, thiazolyl, oxazo the sludge, imidazolyl, indolyl, benzofuranyl and benzothiazole.

Typical alkali metals include sodium and potassium.

Used as sugar substitutes are typically mono-, di - or trisaccharide, or sacharine derivatives, preferably mono - or disaccharides. Preferred are compounds pentoses or hexose. Derivatives include sugar glycosides, N-glycosylamine, O-acyl derivatives, O-methyl derivatives, sugar alcohols, sugar acids, detoxifer or related compounds. Examples include trimethylbenzoquinone hexose compounds IB-00208.

The present invention describes a new polycyclic xanthone-IB-00208 isolated from the fermentation broth of a microorganism, preferably a strain of Actinomadura sp. PO13-046, a culture which was placed in Colección Espanola de Cultivos Tipo University of Valencia, Spain under the access number SISTER 5318. The Deposit of this culture was made in accordance with the provisions of the Budapest Treaty, and any restrictions on the access of the public will be made prior to the issuance of a patent on this application.

The strain of microorganism was isolated from an unidentified marine polychaete worms collected in the Bay of Biscay.

Although depositional microorganism is certainly preferred, the present invention is not tositsa to or not limited to any particular strain or organism. In the scope of the present invention includes other producing compound IB-00208 microorganisms, strains or mutants.

The strain Actinomadura sp. PO13-046, cultivated under controlled conditions in a suitable medium, produces an antibiotic IB-00208. The specified strain preferably grows in an aqueous nutrient medium under aerobic and mesophilic conditions.

Antibiotic IB-00208 can be isolated from sediment of mycelium extraction with a suitable mixture of solvents, such as CHCl3:CH3HE:N2A. the Activity is concentrated in the lower layer. The extracts of the two re-extraction can be combined and one stripped off to dryness in a vacuum.

Isolation and purification of compounds IB-00208 of crude active extract can be performed using a suitable combination of traditional chromatographic methods.

BRIEF DESCRIPTION of DRAWINGS

Figure 1 represents a1H-NMR spectrum of the pure compound IB-00208;

Figure 2 represents a13C-NMR spectrum of the pure compound IB-00208;

Figure 3 is a DEPT experiment purified compound IB-00208;

Figures 4, 5, and 6 represent the COSY 45, HMQC and NMFS spectra of the purified compound IB-00208, respectively;

Figure 7 is an IR spectrum of the pure compound IB-00208;

Figure 8 is an HPLC/MS-chromatogram and ESI-MS spectrum of the joint is IB-00208; and

Figure 9 is a HPLC/UV chromatogram and UV spectrum of the pure compound IB-00208.

The separation into fractions may be conducted on the principle of antitumor activity of fractions, or by TLC with the manifestation of vanillin in concentrated H2SO4or by analytical HPLC with photodiode and MS detector. HPLC analysis was performed at room temperature using an analytical column Symmetry C18 (5 μm)using as mobile phase a mixture of methanol: N2A: acetic acid (99:1:1) with a flow rate of 0.3 ml/min and recording the values at 325 nm, and in these conditions the retention time for compound IB-00208 is 4.5 minutes, as shown in Fig.9.

Based on the detailed analysis of its various spectral characteristics of a pure compound was identified as compound IB-00208 (see the data presented in figures 1-9). Therefore, the established structure corresponds to the one presented above.

These compounds IB-00208 are summarized in Table 1.

Table 1

The absorption maxima of the UV-spectrum: 225, 255 nm and 325 nm, as shown in Fig.9.

1H,13With and DEPT NMR spectra presented in figure 1, figure 2 and figure 3, respectively.

The results of 2D NMR experiments COSY, HMQC and NMFS presented on figure 4, figure 5 and 6, respectively.

The infrared spectrum of popustitelstve 7.

ES-MC-range of detects (M+N)-peak at 691 and (M+Na) peak at 713, as shown in Fig.

Various derivative compounds IB-00208 can also be obtained by applying known in the art techniques. As an example, these changes can be made with the use of chemical and biological methods.

These changes include:

- getting aglionby;

add to the aglycone various known and new sugar;

- modified aglycone.

Can be obtained in connection with various acyl, alkyl, alkenylamine, aryl, benzyl groups and/or groups of sugars in R1. Thus, for example, acylhomoserine and alkyl, alkeline, aryl or benzyl anhydrides can be used as electrophiles for acylation reactions IB-00208 in the relevant basic or acidic conditions in the respective solvents. Alternatively, for alkylation reactions IB-00208 in suitable conditions may apply alkyl-, alkenyl-, aryl - or benzylchloride, mesylates or tozilaty. Additionally, various derivatives of sugars, known and new, can be used as reagents for the reactions glikozidirovaniya under standard conditions for this type of reaction.

On the other hand, compounds containing hydrochinone ring,can be obtained from compound IB-00208 recovery quinone suitable regenerating agent [for example: NaBH 4(.Ross Kelly et al., J. Am. Chem. Soc., 1989, 111, 4522-4524), Na2S204(A.V.Rama Rao et al., Tetrahedron Lett., 1991, 32, 5199-5202)] in a suitable solvent, followed (when R1≠H) conducting interaction hydroquinone with allermuir or or suitable alkylating agent in a suitable solvent. As described above, various acyl, alkyl, alkeline, aryl, benzyl and/or groups of sugars can be implemented in R1. Also different alkyl groups can be introduced in R2and R3. In addition, R2and R3can be connected by bonds of, for example, by reaction with intramolecular metathesis circuit loop between the two terminal double bonds.

The present invention with reference to the preferred embodiments below further illustrates by examples, which are intended to assist in understanding the present invention and to illustrate it, and should not be construed as limiting it.

EXAMPLES of INVENTIONS

All these are percentages, unless otherwise stated, is presented on weight. All these temperature expressed in degrees Celsius. All incubation was carried out at 28°unless otherwise stated, and the flasks were shaken in a circular shaker with a speed of 250 rpm All environments and the recipients were sterile, and all operations of the aseptic cultures.

Examples 1-3 relate to compounds IB-00208 above formula. Examples 4-9 relate to the formation of compounds 1-6 according to the invention. Example 10 relates to the determination of biological activity of compounds IB-00208.

EXAMPLE 1

Producing microorganism.

Used here taxonomic methods are methods that are usually used in classical taxonomy Actinomycete, and described in the literature.

All cultures were examined after incubation, and all records of the results was done on a weekly basis up to 21 days. If necessary NaCl was added.

Description of the microorganism is as follows.

After 21 days there has been good growth in the colonies ISP 2 ISP 6, Bennet and 172 of ATSS with ASW. The colony had a light brown color. In the colonies ISP 3 ISP 5, Czapek and ISP 7 ASW was observed worst growth and soluble pigment has not been detected. Aerial mycelium is not formed. Substrate mycelium was extensive. On substrate mycelium could be formed isolated disputes. Disputes were elongated and rare. No other formations were not observed.

In the colony ISP-1 and other hard materials, is formed capable of diffusion brown pigments. Resistance to NaCl than 5%. The optimal growth temperature range was from 25° to 35°C. the Microorganism was able to grow on glucose, galactose, is amnote and xylose as the sole carbon source, however, growth on fructose, raffinose, m-Inositol, sucrose and α-melibiose was not observed, and mannitol and melezitose was hesitant.

Studies of the chemical composition of the microorganism showed that the hydrolyzed whole cells of strain RU-046 contains meso-2,6-diaminopimelic acid.

Comparison of methyl esters of fatty acids of strain RU-046 with other similar strains described in Table 2.

Based on the foregoing characteristics, culture was identified as a species of the genus Actinomadura, with 94% identity to strain A. vinacea.

EXAMPLE 2

Fermentation IB-00208:

The steps required for producing the compounds IB-00208 preferred microorganism, are:

Source culture: Full broth of strain R-046 pure culture of Actinomadura sp. frozen in 20% glycerol.

Sowing culture: the Frozen culture or growing culture on the sloped agar was used for inoculation of 100 ml of the previously described seed medium in 250 ml shake flask. The flask is incubated for 48 hours and used as a seed culture of the first stage. 500 ml of the same medium in a two-liter Erlenmeyer flask were seeded at 10% of the specified seed culture of the first stage. The flask is incubated for 48 hours.

Fermentation: 75-lit the new tanks for fermentation 50 liters already described production medium was seeded with 2.5 liters of the seed culture of the second stage. Fermentation was carried out for 96 hours under stirring at a speed of 400 rpm in a stream of air 0,5/oben

Table 3
The sowing medium
Glucose5 g
Starch20 g
Meat extract3 g
Yeast extract5 g
Triptan5 g
Caso34 g
NaCl5 g
KCl0.5 g
MgCl22 g
Bringing waterto 1 liter
Production environment
Glucose5 g
Starch20 g
Soy flour15 g
Yeast extract5 g
Triptan2 g
Caso34 g
NaCl5 g
KCl0.5 g
MgCl22 g
Bringing waterto 1 liter

Output connection IB-00208 was assessed by determining activity against leukemia mice P-388 full broth or by HPLC.

EXAMPLE 3

The selection of compounds IB-00208.

the 4.5 litre full of cultured broth was filtered to separate the biomass and other solids. The precipitate of mycelium were extracted twice with 1.5 l of a solvent mixture of CHCl3:CH3HE:N2O (2:1:1), the activity was concentrated in the lower layer. The organic solvent was concentrated and evaporated to dryness in vacuum to obtain 1.2 g of the crude extract.

The extract was separated chromatographically on silica gel, elwira mixtures of n-hexane/ethyl acetate and ethyl acetate/methanol. 110 mg of the fraction containing possessing antitumor activity of compound IB-00208, suirable with ethyl acetate/methanol (1:1).

Further purification containing compound IB-00208 fractions was performed by the method of column chromatography on silica gel and was suirable chloroform/methanol (95:5) 18 mg of pure compound IB-00208.

Example 4

Getting Connection 1:

The solution IB-00208 (150 mg, 0.22 mmol) in toluene (15 ml) is treated with Ag2O (183 mg, of 0.79 mmol) and allylbromide (0,76 ml) at room temperature in argon atmosphere. The reaction mixture is stirred for 5 hours at 70°C. After cooling to room temp the atmospheric temperature the mixture is filtered through celite® and washed with dichloromethane. The solvent is evaporated under reduced pressure and the residue purified flash chromatography (CHCl3:Meon 10:0,4) to obtain the corresponding pure product (yield 34%).

Example 5.

Getting Connection 2:

A solution of Compound 1 (10 mg, 0.014 mmol) in a mixture of methanol/dichloromethane 9:1 (3 ml) is treated with an aqueous solution of 10 M H2SO4(0.5 ml). After cooling to room temperature the mixture was diluted with CH2Cl2and add saturated aqueous solution of NaCl. The aqueous phase is extracted with dichloromethane and dried over anhydrous sodium sulfate. After evaporation of the solvent under reduced pressure and flash chromatography (CHCl3:Meon 10:0,4) get a pure Compound 2 (yield 30%).

Example 6.

Getting Connection 3:

The solution IB-00208 (50 mg, 0,072 mmol) in dichloromethane (0.7 ml) is treated with palladium on coal (10 mg Pd/C) and stirred at room temperature in a hydrogen atmosphere for 4 hours. The mixture is filtered and the solvent is evaporated under reduced pressure. The corresponding recovered product with hydrogenosomes fragment is received with a quantitative yield (100%).

Example 7.

Getting Connection 4:

To a solution of Compound 3 (169 mg, 0.24 mmol) in dichlor the Tanya in an argon atmosphere at 0° To add 0,02 ml of pyridine (0.24 mmol). The reaction mixture was stirred at 0°C for 1 hour and add 0.2 equiv of pyridine and 0.2 EQ of acetic anhydride to the total number of 1 EQ. The reaction is accompanied by TLC, and then the reaction mixture was poured into ice. pH adjust up to 1-2 using 1M HCl and the reaction mixture is extracted with dichloromethane. The organic phase is washed with saturated aqueous NaCl and dried over anhydrous sodium sulfate. After evaporation of the solvent under reduced pressure and flash chromatography receive net monoacetylmorphine compound Compound 4 (yield 87%).

Example 8

The Connection 5:

To a solution of Compound 3 (92 mg, 0.133 mmol) in pyridine in an argon atmosphere at room temperature, add 0.4 ml of Et3N (2,9 ml) and 0.64 ml of acetic anhydride (to 6.78 mmol).

The reaction mixture was stirred at room temperature for 1 minute and then add crushed ice. The mixture is then poured into a solution of 1 M HCl at 0°and extracted With dichloromethane. The organic phase is washed with saturated aqueous NaCl and dried over anhydrous sodium sulfate. After evaporation of the solvent under reduced pressure and flash chromatography receive net triazacyclononane Compound 5 (yield 74%).

Example 9.

The Connection 6:

<>

Use the same technique as described above for Compound 5. In this case, the reaction mixture is stirred for 5 minutes before processing to obtain deacetylating product of Compound 6 as the primary connection.

EXAMPLE 10

The connection 15 and compound 16

The solution IB-00208 (18 mg, or 0.027 mmol) in THF (1 ml) of NaH 60% (1,16 mg 0,029 mmol) at 0°C in argon atmosphere. The reaction mixture is stirred for 5 minutes at this temperature. The solution becomes green, and the formed solid substance (sodium salt, compound 15).

Then at 0°With add benzylbromide (3,5 μl, 0,029 mmol) and tetrabutylammonium iodide (1.1 mg, of 0.003 mmol). The cooling bath removed and the reaction mixture is stirred for 4 hours. Quenching of the reaction carried out by addition of 0.1 n HCl. The mixture is then extracted with dichloromethane (twice). The combined organic extracts are dried over bezw. Na2SO4, filtered and the solvent is evaporated under reduced pressure. The connection 16 see in trace quantities using the1H-NMR of the crude product.

The connection 16:

The solution IB-00208 (18.5 mg, 0.012 mmol) in toluene (15 ml) is treated with Ag2O (12 mg, 0.05 mmol) and benzyl what romida (6 μl, 0.05 mmol) at room temperature in argon atmosphere. The reaction mixture is stirred for 6 hours at 70°C. After cooling to komnatnoi temperature, the mixture is filtered through celite® and washed with dichloromethane. The solvent is evaporated under reduced pressure and the residue purified flash chromatography (CH2Cl2:Meon 100:1) to give the pure compound 15 (5 mg, yield 52%) as an orange solid.

EXAMPLE 11

Biological activity.

The antitumor activity of compound IB-00208 was determined in vitro in cell cultures of leukemia mice P-388, the human lung carcinoma A-549, carcinoma human colon HT-29 and human melanoma MEL-28. The procedure was performed according to the method described in Bergeron et al. It was shown that the IC50 value for all 4 cell lines cells were 0,001, of 0.001, 0.001 and 0.001 microg/ml, respectively. Similar results were obtained by the same method for compounds 1-6.

LINKS

In the description of the present invention includes the following links:

ATSS 1996 Catalog;

Bergeron et al., Biochem. Biophys. Res. Comm., 121: 848, 1984;

Guerrant and Moss, Anal. Chem., 56: 633, 1984;

Hasegawa et al., J. Gen. Appl. Environ., 29: 319, 1983;

Shirling and Gotlieb, Int. J. Syst. Bacterial., 16: 313, 1966;

Van der Auwera et al., J. Environ. Methods, 4: 265, 1986;

Waksman, The Actinomycetes, vol.II: 331, 1961.

The present invention has been described in detail, including its preferred embodiments. Od is ako clear the person skilled in the art based on the description of the present invention may make changes and/or improvements that are not beyond the scope of the present invention.

1. The compound of General formula:

or

where each substituent R1is the same or different and can represent hydrogen, acyl, alkyl, alkenyl, aryl, benzyl, alkali metal and/or a monosaccharide, a disaccharide, trisaccharide or their derivative, a R2and R3can represent hydrogen or alkyl.

2. Compound IB-00208 according to claim 1 of the formula:

3. The method of obtaining the compounds IB-00208, which includes the cultivation of a strain of microorganism capable of producing in an aqueous nutrient medium.

4. The method according to claim 3 which further includes isolation and purification of compounds IB-00208 from the culture broth.

5. The method according to claim 4, where producing the compound IB-00208 microorganism is an essentially pure strain culture RE-046, available under the access number SISTER 5318 in Coleccion Espanola de Cultivos Tipo University of Valencia, Spain.

6. Essentially pure strain culture RE-046, which produces the connection IB-00208 isolated from marine invertebrates, C is asih to the family Thermomonosporaceae and taxonomically classified as Actinomadura sp.

7. Antitumor pharmaceutical composition comprising a compound according to claim 1 and a pharmaceutically acceptable carrier.

8. Antitumor pharmaceutical composition comprising a compound IB-00208 according to claim 2 and a pharmaceutically acceptable carrier.



 

Same patents:

The invention relates to the field of biotechnology and Microbiology

The invention relates to biotechnology and relates to a new and improved method of allocation of clavulanic acid from the aqueous culture broth producer clavulanic acid

The invention relates to a method of stereoselective recovery phenylalkylamines to their respective (S)-hydroxycodone and the new strain

The invention relates to biotechnology, namely microbiological synthesis and release from biomass actinomycete Streptomyces avermitilis avermectins used for antiparasitic drugs

The invention relates to Microbiology and relates to a new strain actinomycete for the production of antiparasitic drugs on the basis of avermectins
The invention relates to the field of biotechnology, namely, to receive the berberine in immobilized cell culture vasilistnika small (Thalictrum minus L.)

FIELD: medicine, in particular, laboratory-based methods for diagnosing tuberculosis.

SUBSTANCE: into diagnostic material, after processing with detergent - chlorhexidine gluconate prior to seeding in dense nourishing substances additionally injected is freshly prepared water solution of hydro-gel of methyl-silicon acid in ratio 2:1, shaken, centrifuged, to precipitation 1 ml of special substance is added, product is taken to dense nourishing substance.

EFFECT: method accelerates growth of tuberculosis mycobacterium and accumulation of bacterial mass, which is important for differential diagnosis of tuberculosis.

2 tbl

FIELD: biotechnology, in particular methods for biomass flocculation from suspending media.

SUBSTANCE: claimed method includes addition the first cationic polymer material into suspending medium, wherein said material has intrinsic viscosity not more than 2 dl/g, and simultaneous or subsequent addition of the second cationic or substantially non-ionic polymer material into suspending medium, wherein said material has intrinsic viscosity of at least 4 dl/g. Said dosage provides optimum characteristics for biomass flocculation. Method of present invention makes it possible to obtain flakes with increased endurance, and as a result to apply various isolation methods including those based on mechanical flake agitation.

EFFECT: improved method for biomass flocculation from suspending media.

15 cl, 4 tbl, 6 ex

FIELD: biotechnology, veterinary science.

SUBSTANCE: invention relates to the strain that is deposited in the Collection of strain-pathogens of diseases in fur animals in the Museum of bacterial and viral structures of the Science-Research institute of fur farming and rabbit farming (NIIPZK) named for V. A. Afanasiev and has the registration number 71. The strains relates to streptococci of the group B. The strain is cultures in Todd-Hewite beef-extract broth, Pike medium at 37°C for 18-20 h. After control of the grown culture for virulence the separated biomass is inactivated by addition of 0.5% formaldehyde followed by addition of aluminum hydroxide as an adjuvant and packaging. Invention provides preparing the effective vaccine against streptococcosis in fur animals based on this strain.

EFFECT: valuable properties of strain.

1 ex

FIELD: biotechnology, veterinary science, microbiology.

SUBSTANCE: strain is prepared by fusion of E. coli genes isolated from mink intestine that induce biosynthesis of microcine C51 and B-subunit of escherichia thermolabile enterotoxin into the genome. Microcine inhibits growth and development of enterobacteria producing thermolabile, thermostable and shiga-like (verotoxin) enterotoxins, and B-subunit screens intestine receptors for enterotoxin and initiates specific immunity in intestine as far as intestine comprises determinants of protective antigen of toxin molecule. The strain is used for preparing curative-prophylactic preparations with prolonged action. Microcines synthesized by the strain inhibit enterotoxic enterobacteria and synthesized B-subunits of escherichia thermolabile enterotoxin screen receptors for enterotoxin and stimulate specific immunity in intestine. Resident properties of a carrier don't cause rejection of the strain by the mink intestine microecosystem that allows applying these preparations based on thereof more rarely and in smaller doses.

EFFECT: valuable properties of preparation.

2 cl, 4 ex

FIELD: microbiology, pharmacy.

SUBSTANCE: invention relates to a method for isolating biomass of microorganisms from cultural fluid and can be used in manufacturing preparative formulations of biopreparations. Method involves addition of high-molecular reagent "Givpan" as a flocculant representing a product of hydrolysis of polyacrylonitrile copolymer of acrylonitrile with methacrylate. "Givpan" is added in the amount 0.1-0.8 wt.-% of the cultural fluid volume. Applying the proposed method allows retaining activity of microorganisms to be isolated that are antagonists of fungal phytopathogens and amount of viable cells.

EFFECT: improved isolating method.

3 tbl, 3 ex

The invention relates to Mycology

The invention relates to biology and medicine

The invention relates to Microbiology and can be used to highlight axenically cultures of microalgae at the physiological, biochemical, cytological and genetic analysis of microscopic algae

The invention relates to the production technology of medical immunobiological preparations, in particular, to a method of concentrating spore cultures in the production of the anthrax vaccine, ensuring the stability of their biological properties with preservation of potency, and can be used in the practice of production of the anthrax vaccine

The invention relates to the field of medicine - pharmacology, and more specifically to methods for selection of antibiotics aminoglycosides

FIELD: pharmaceutics.

SUBSTANCE: the suggested composition includes platinum-based compound with spatially complicated coordination and not platinum-based anti-cancer agent, and pharmaceutically acceptable carrier or solvent; the innovation deals with their application for obtaining medicinal preparation for inhibiting the growth of tumor cells in such a patient; with the method to inhibit the growth of tumor cells due to introducing for a patient efficient quantity of the mentioned therapeutic combined product or pharmaceutical composition that inhibits the growth of tumor cells according to the present innovation; in particular, when platinum-based compound with spatially complicated coordination is being (SP-4-3)-(cis-amine dichlorine-[2-methylpyridine]platinum (II) or its promedicine one should choose not platinum-based anti-cancer agent out of taxol, haemcytabin, navelbin, doxyl, 5-FU and taxoter. The present innovation enables to overcome resistance to cisplatin and prevent platinum-associated toxicity.

EFFECT: higher efficiency.

18 cl

Up!