Tuamarina, possess antibacterial properties, the microorganism strain alteromonas rava sank-73390 - producer thimerisol and a method of producing antibiotic thimerisol


(57) Abstract:

Usage: as an antibacterial agent. Tuamarina obtained by cultivation of a strain of Alteromonas rava SANK 73390 and produce the target product from the culture fluid. 3 C. p. F.-ly, 3 tables.

The invention relates to a new connection, which is called "Thiomarind", as well as the method of preparation thimerisol, including enzyme using microorganism biological genus Alteromonas and especially the model of a new species of Alteromonas rava marked SANK 73390, which is novel in itself and is part of the present invention. Tuamarina possesses many curative, in particular antibacterial, effects, therefore, the invention also provides compositions and methods for therapeutic or prophylactic use this connection.

Organisms of the genus Alteromonas can be extracted from sea water and it was shown that they form compounds suitable for therapeutic use. For example, a compound known as Viscabelin was obtained from one of the species of Alteromonas and it was shown that it elicits antitumor activity (Japanese patent Kokai application N Sho 63-27484).

What concerning the society can be divided into three groups.

The first group includes pseudomonaceae acid primarily secreted from Pseudomonas species. These compounds include Pseudomonas acid And [formed by Pseudomonas fluorescens disclosed in J. Chem. Soc.Perkin Trans, 1, 294, 1977] Pseudomonas acid [ibid 2827, 1982] and Pseudomonas acid D [ibid, 2655, 1983] Pseudomona acid And sold under the name "Bactroban" (registered trademark Beleham) in the form of 2% dermatological ointments for antibacterial use.

Other derivatives Pseudomonas acid were obtained from marine bacteria [Am. Chem. Soc. Abstr.Pap, 200 (2), (1990)] but this link does not issue any bacterial activity.

The second group of substances having a structure similar to the structures of the compounds of the invention, includes the group that contains the antibiotic colomycin [Helv. Chim. Acta, 42, 563, 1959] pyrrhotite [J. Am.Chem.Soc. 77, 2861, 1955] thiolutin [Anero. Chem. 66, 745, 1954] aureolin [J. Am.Chem.Soc. 74, 6304, 1952] and others. These antibiotics are formed typical radiant fungi and characterized by the presence serosoderjaschei chromophore. Xenorhabdus I-V are substances related to golomazin, and also can be isolated from bacteria (described in WO 84/01775).

Conducted various research progenie thimerisol or which would be characterized by similar properties.

The third group of compounds disclosed in publications such as Japanese patent application Kokai NN 52-102279, 54-12375, 54-90179, 54-103871 and 54-125672, which reveal pseudomonaceae acid derivative, having a structure similar miamarina, but in which the terminal carboxylic acid substituted on the amino group. These compounds do not reveal comparable antibacterial activity and do not show a wide spectrum of antibacterial activity. Indeed, these compounds tend to have more weak antibiotic activity compared to the original Pseudomonas acid.

None of the above compounds was isolated from species of Alteromonas and none is identical miamarina. For example, one of the characteristics of the structure thimerisol is the presence of Oh-groups located between the 6-membered ring and ,-unsaturated carbonyl group. Accordingly, tuamarina has a distinct difference from those described in the prior art.

The aim of the invention is to provide new compounds having improved efficacy and broader spectrum of antibacterial activity in comparison with the above-described groups of antibiotics.

Sledovatelnot thimerisol, which includes raising teamarena-forming microorganism of the genus Alteromonas and selection thimerisol of culture.

The invention also provides a pharmaceutical composition comprising tuamarina in a mixture with a pharmaceutically acceptable carrier or diluent.

Hereinafter the invention is the use of thimerisol in therapy, in particular for the treatment or prevention of bacterial infections.

The invention provides for the use of thimerisol for the manufacture of drugs for the treatment or prevention of bacterial infections.

In addition, the invention provides a method of treatment or prevention of bacterial infections, which includes receiving an effective amount of thimerisol mammal, which may be human, suffering from such infection or sensitive to it.

From the foregoing it is clear that tuamarina contains a number of asymmetric carbon atoms and more double bonds. Isomerization, in particular, possible to ,-unsaturated carbonyl part thimerisol. Thus, tuamarina can form various geometrical and optical isomers. And although all these compounds are represented here UOM mixtures thereof, including the racemates. When using stereospecific synthesis techniques or optically active compounds as starting substances can be received directly by the individual isomers; on the other hand, if preparing a mixture of isomers, the individual isomers may be obtained using standard methods of separation.

Natural tuamarina tends to take a standard optical configuration. Thus, although other configurations and provides, preferred is a natural configuration.

Tuamarina can be obtained by growing timeresolution a microorganism of the genus Alteromonas and then collecting thimerisol from the culture medium. Options thimerisol with the necessary antibacterial activity can be obtained similarly from other species or samples Alteromonas, which form the desired connection, or they can be obtained by appropriate modification of the compounds obtained by fermentation as described, or they can be synthesized by chemical means.

In particular, we especially prefer to use as the new microorganism samples Alteromonas rava-Anisman, which was isolated from seawater collected in the area of sea Koina, Minamiyzu Machi, Prefecture Shiruona, Japan, and this species was deposited in the Institute of Escrow, Research Institute of microbial technology, Agency of industrial research and technology, Japan, April 30, 1991 N registration FERM BP-3381 according to the Budapest Treaty.

Taxonomie characteristics of the species Alteromonas rava SANK 73390 presented below.

1. Morphological characteristics.

View Alteronomas rava were grown at 23oC for 24 h on marine agar (Difco). Subsequent microscopic observations showed that the cells had Blockoban shape and each have a size in diameter from 0.8 to 1.0 microns and in length from 2.0 to 3.6 μm. This species is gram-negative and develops through the polar odnochastichnogo flagellum.

2. Growth on marine agar.

SANK 73390 were grown for 24 h at 23oC on marine agar (Difco). The resulting colonies were observed in the form of pale-yellow gray-yellow, round, flat and solid formations. Water-soluble pigment is not formed.

3. Physiological properties.

(1) Requirements for sea water: SANK 73390 requires for growth seawater. the prepared from artificial sea water): no effect on carbohydrate.

(3) Oxidase: +

(4) Catalase: +

(5) Oxygen requirement: aerobic

(6) Reduction of nitrate: -

(7) Hydrolysis of starch: +

(8) Decomposition of agar: -

(9) Liquefaction of gelatin: +

(10) the Development ol Naze:

(11) Production of lipase: +

(12) Temperature for growth: weak growth at 4oC good growth between 17oC and 26oC, no growth at 35oC.

(13) the Requirement for growth factor: basic nutrient media described in Journal of Bacteriology, 107, 268-294 (1971), SANK 73390 requires not contain vitamin Casamino acid.

(14) Assimilation of carbon sources: the main nutrient media described in Journal of Bacteriology 107, 268-294 (1971), further including 0.1 wt. the volume does not contain vitamin Casamino acid under cultivation in the conditions of mixing (see tab. 1).

4. Chemotaxonomical feature.

(1) Mol. guanine and cytosine (G + C content of DNA: 43,4% (HPLC method).

(2) Quinone system: Ubiguinon Q-8.

Taking into account the above taxonomic characteristics of the species Alteromonas rava SANK 73390 were compared with the species described in Bergey''s Manual of Systematic Bacteriology, I. 1 (1984), and also with similar kinds described in the latest editions of Midhe Alteromonas citrea, other marine organism. SANK 73390 and Alteromonas cirea ADS 29719 (standard view) was cultivated in the same conditions and compared.

Compared with yellow, gray-yellow microorganism SANK 73390, the colony was ATSS 29719 were greenish-yellow color. SANK 73390 differs from Alteromonas citrea the growth rate at 4oC and the ability to assimilate trehalose and sodium propionate as carbon sources.

Accordingly Alteromonas rava views SANK 73390 is a new species Alteromonas rava and different in basic characteristics from the nearest known species, selected in accordance with the directory N ATS 29719.

The above characteristics are typical characteristics of the species SANK 73390. However, it is well known that the characteristics of Alteromonas species are variable both for natural and artificial kinds. The above characteristics determine the selected species Alteromonas rava, but are not typical of other species of Alteromonas or species Alteromonas rava, which are capable of forming tuamarina or existing in nature varieties. Such other types fall under the scope of the invention.

It is obvious that SANK 73390 or any other type of microorganism capable of producing the who change or modification with the formation of the body with different characteristics. The only requirement is the requirement that the formed body was capable of producing the desired connection.

Such alteration and modification can take any desired shape, or may be due to such phenomena as, for example, culturing conditions. Types can be changed by cultivation and in this selection, which aims to create the characteristics of growth or growth at a lower/higher temperatures.

The target can be mainly biotechnological modification and can be represented by selected characteristics, such as bacteriostatic resistance or susceptibility, or their combination, in order to preserve the purity or to create an opportunity for purification of crops, especially crops from time to time.

Other characteristics that can be produced by genetic effects, are any characteristics that are valid for species Alteromonas. For example, can be combined resistance plasmid encoding or can be removed any existing in nature plasmids. Favorable plasmids are those that provide auxotrophy properties. Plasmids can be is in the nature of the plasmid Altromonas and the introduction of the desired gene or genes from another source. Natural plasmids can also be modified in any other desired way.

In order to get tuamarina from the culture of the respective microorganism, microorganisms must be fermented in an appropriate environment. These environments are generally well known and are often used when creating other enzymatic products.

Typical is, when the nutrient medium is a need to have any combination of carbon source, nitrogen source and one or more inorganic salts, assimilated the corresponding microorganism. The minimum requirement for a nutrient is the requirement that it contains those ingredients that are essential for growth of the microorganism.

Relevant carbon sources are glucose, fructose, maltose, sucrose, mannitol, glycerol, dextrin, oat food, grain vodka, corn starch, potatoes, corn powder, soybean powder, cottonseed oil, syrup, citric acid and tartaric acid, any of which may be used alone or in combination with another or with a large number of these compounds. Typical use kolichestvennie be changed in accordance with the desired result.

Appropriate sources of nitrogen are any compounds containing protein. Examples of nitrogen sources are organic sources of nitrogen of animal and vegetable origin, they can also be extracts from such natural sources as soybean powder, powder gorokhovich grains, powder of cotton seeds, casein hydrolysate, feramin, fish powder, grain leached liquid, peptone, meat extract, malt extract, and from such inorganic sources of nitrogen as sodium nitrate, ammonium nitrate and ammonium sulfate. As carbon sources, they can be used alone or in combination with each other. Typical appropriate number of such substances may be the interval from 0.1 to 6 wt. from the volume of the nutrient medium.

Appropriate nutrient inorganic salts are those that provide trace elements and major alternates salts. Preferably the salt must provide such ions as sodium, potassium, ammonium, calcium, magnesium, iron, phosphate, sulfate, chloride and carbonate. May be traces of metals such as cobalt, manganese, and strontium, or salt, is able to give ions such as bromide, fluoride, O in the absence of contrary indications of the conditions for the development of its culture ideally suited to a marine environment. Therefore, traces of ions found in the sea, have a beneficial effect when included in any medium used for the cultivation of Alteromonas.

In the case where the microorganism is fermented in the form of liquid culture, preferred is the use of anti-foam agent such as silicone oil or other suitable surfactant.

Preferably, the pH of the culture medium for Alteromonas rava views SANK 73390 when used for the production of thimerisol was maintained at pH from 5.0 to 8.0, although the only requirement is that the pH value did not prevent the growth of microorganism or no adverse irreversible effects on the quantity of the finished product. It is preferable to add an excess of acid or alkali to stop fermentation.

Alteromonas rava views SANK 73390 mainly gives rise in the temperature range from 4 to 32oC, and grows well at a temperature of from 17 to 26oC. Can be used, and other temperature outside this interval, when there is a species that can grow at lower or higher temperatures. For the production of thimerisol preferred temperature of the culture and can be used in any appropriate way aerobic cultivation, such as solid cultivation, cultivation using mixing or aeration-agitating cultivation.

If the cultivation is carried out on a small scale, then enzyme cultivation using the mixing is carried out preferably for several days at a temperature of from 20 to 26oC.

At the beginning of the enzymatic cultivation preferably use the method of initial seeding made in one or in two stages, for example, in the Erlenmeyer flask. For cultural nutrient medium may be a combination of carbon source and nitrogen source. Sowing flask potrahivaya in a cooled incubator at a temperature of 23oC for 1-3 days or until until you experience sufficient growth of organisms. The resulting sowing culture can be then used for inoculation of the secondary seed culture or productive culture. If the second planting, it is similar and partially used for inoculation of the nutrient medium. Bulb, in which the sowing, stirred for 1-3 days or until until is reached Multivitamine the contents of the flask may be collected by centrifugation or filtration.

If the cultivation is carried out on a large scale, preferably used in a suitable culture aeration-mixing the fermenter. When using this method, the nutrient medium can be prepared in the fermenter. After sterilization at 125oC environment is cooled and is planting the appropriate seeds, pre-grown in a sterilized environment. The cultivation is carried out at a temperature from 20 to 26oC under stirring and aeration. This method is suitable for obtaining a large number of connections.

The number thimerisol formed by cultivation, can be monitored over time, for example, high performance liquid chromatography. Usually the number of educated thimerisol reaches its maximum after a time from 19 to 96 hours

Upon reaching the appropriate period of time tuamarina can be isolated and purified by any known method. For example, tuamarina remaining in nutrient broth may be obtained, for example, by filtering solids using diatomaceous earth as a filtration media or by centrifugation and subsequent extractee is, is mausica in the filtrate or in the pop-up layer can be extracted miscible with water, an organic solvent such as ethyl acetate, chloroform, telengard, methylene chloride or any of their mixtures in a neutral or acidic environment cleaned up.

Or as adsorbent can be used activated carbon or adsorbent resin, such as Amberlite XAD-2, XAD-4 (Rohm Haas) or Diaion HP-10, HP-20, SER-20, HP-50 (Mitsubishi Kesei Corporation). Impurities can be removed after adsorption by passing the liquid containing tuamarina, through the layer of adsorbent; or tuamarina can be cleared after adsorption by elution with a suitable eluent such as aqueous methanol, aqueous acetone or butanol/water.

Intracellular tuamarina can be purified by extraction with a suitable solvent, such as 50-90% aqueous acetone or aqueous methanol, followed by removal of organic solvent, accompanying the extraction, as described above for the filtrate or pop-up layer.

Formed tuamarina can be further purified well-known methods, for example by passing through the adsorption chromatographic column using a carrier such as silica gel or through the column using as adsorbent such substance, as Sephadex ZH-20 (trade name for a pharmaceutical product); high performance liquid chromatography using a column of the normal phase or reverse phase. As is known, such methods of isolation and purification can be conducted individually or in any combination, repeatedly if necessary, to isolate and purify the desired final product.

In the case where compounds of the invention are used for therapeutic purposes, they can be taken by themselves or in an appropriate pharmaceutical product containing in addition to the active compounds, one or more standard diluents, carriers, fillers or excipients. The nature of the pharmaceutical composition, of course, depends on the intended route of administration. However, for the oral route of administration compound is preferably prepared in the form of powders, granules, tablets, capsules or syrups. For parenteral receive preferred is the preparation in the form of injections (which can be administered intravenously, intramuscularly or subcutaneously) or in the form of drops or suppositories.

The preparations can be prepared with known methods and with the addition of such to luchshaya substances, solubilizing additives, suspensorysex agents, or substances used for coating. Although the dosage may vary depending on the symptoms and age of the patient, nature and severity of the infection and the path and method of reception in the case of oral administration to adult humans of the compounds of the invention, taken generally in the form of a daily dosage of from 20 to 2000 mg of the Compound can be taken as a single dose or divided doses, for example two or three times a day.

Tuamarina shows antibacterial effect against gram-positive and gram-negative bacteria in animals (e.g. humans, dogs, cats and rabbits), often relevant is its use usually in the form of cream, ointment or gel. Similar consideration in the compilation and preparation of such forms of acceptance similar to the data above.

The following examples illustrate the obtaining thimerisol and its antibacterial activity, but they should not be construed as limiting the invention in any way.

Example 1. Fermentation thimerisol in the fermenter.

A) Culturing.

Alteromonas rava strain SANK 73390 was cultivated for 3 days at 22oC on stubble is spencie was taken aseptically and inoculates in a 500 ml Erlenmeyer flask, containing 100 ml of sterilized nutrient medium [37,4 g marine broth (Difco) in 1 l deionized water, pH was not regulated]

The flask was thermostatically under stirring for 24 hours at 23oC at 200 rpm (rotation radius of 70 mm) using a rotary shaker. After this time in each of the four 30-liter fermentors, each containing 15 liters of sterile nutrient medium, inoculable 15 ml culture was taken aseptically from the Erlenmeyer flask. The fermenters were thermostatically under stirring (100 rpm./min) for 23 h at 23oC and at a speed of aeration 7.5 l/min

In The Selection.

After 23 h the contents of the digester was connected with the output 60 l of liquid culture. Then the pH of the liquid was regulated to a value of 3 by the addition of hydrochloric acid, followed by adding 60 l of acetone and the mixture was extracted for 30 min with stirring. The solution was filtered using a 1.2 kg SelidovUgol filtration media 545 (Trade mark of a product obtained from John Manville Co.). Then 110 l of the resulting filtrate was extracted with 60 l of ethyl acetate once and twice more with ethyl acetate each time 30 L. the Organic layer was washed 30 l IDA sodium, were dried over anhydrous sodium sulfate and supariwala until dry under reduced pressure with the formation of 14 g of oily substance.

The obtained oily substance was dissolved in methylene chloride and the solution was subjected to adsorbirovanny on a column filled with 200 g of silica gel in methylene chloride. The target compound was subjected to elution with solvents of increasing polarity in the following order: methylene chloride/ethyl acetate; ethyl acetate; ethyl acetate/methanol. Eluent was collected into fractions in 18 ml fractions, erwerbende a mixture of ethyl acetate-methanol, which contained tuamarina, was located.

Caught fraction was condensed to dry by evaporation to obtain 7 g of an oily substance, which was dissolved in 400 ml of 50% aqueous bulk amounts of water and methanol was adsorbiroval 600 ml column filled with Diaion HP-20 (trademark of the product obtained from Mitsulishi Chem. Jnd. ) in water. After washing 50% by number by volume aqueous solution of methanol, the target substance was subjected to elution with 90% of the amount by volume of methanol and water after condensation until dry under reduced pressure there was obtained 1 g of yellow powder. Yellow powder art with ethyl acetate and methanol (19: 19:2 by volume) to collect active fractions. Received 750 mg thimerisol in the form of a yellow powder.

The resulting Tamarina had the following properties.

1) the Nature and appearance: yellow powder

2) melting Point: 84-89oC

3) Molecular formula: C30H44N2O9S2< / BR>
4) Molecular weight: 640 determined by the method of "FAB-MS (mass spectrometry method bombardment of heavy atoms).

5) Mass spectrometry high resolution:

WITH30H45N2O9S2[(M+N)+according to the method of bombardment by heavy atoms]

Calculated: 641,2567

Found: 641,2585

6) Elementary analysis:

Designed WITH 56,23; N 6,92; N 4,37; S 10,01;

Found, 55,92; N 6,82; N 4,23; S 9,90.

7) Infrared absorption spectrum: the infrared spectrum shows the following peaks (using KBr disk, nmaxcm-1):

3394, 2930, 1649, 1598, 1526, 1288, 1216, 1154, 1102, 1052.

8) Ultraviolet absorption spectrum:

In methanol or methanol + HCl tuamarina has an ultraviolet absorption spectrum, shown below: [given asmaxnm ()]

387 (12,000), 300 (3,500), 214 (26,000) and in methanol + NaOH has a UV spectrum, shown below: [this as ]

386 (9,600), 306 (3,200), 206 (25,000).

Separation column: Senshu - Pak ODS H-2151. (Column size h mm, a Product of Senshu Scientific Co. Ztd.).

Solvent: 40% by volume acetonitrile.

The flow rate: 1.5 ml/min

Wavelength: 220-350 nm (defined next photodiode).

Retention time: 5,9 min

11)1H-spectrum nuclear magnetic resonance: (mil.D.). Spectrum of nuclear magnetic resonance (270 MHz) hexadeuterated dimethyl sulfoxide using as an internal standard tetramethylsilane shown below:

of 0.91 (3H, doublet, J=6,8 Hz);

of 0.95 (3H, doublet, J=5,9 Hz);

1,30 (6N, broadened, multiplet);

of 1.55 (5H, broadened, multiplet);

2,03 (3H, singlet);

of 2.09 (3H, multiplet);

of 2.34 (2H, triplet, J=7,3 Hz);

3,3 (1H, doublet, J=10,7 Hz);

to 3.52 (2H, multiplet);

of 3.64 (2H, multiplet);

to 3.73 (1H, double doublet);

as 4.02 (2H, triplet, J=6.6 Hz);

4,18 (1H, broadened. doublet, J=7,3 Hz);

4,30 (1H, doublet, J=4.4 Hz);

of 4.44 (1H, doublet, J=7,8 Hz);

4,63 (1H, doublet, J=3,4 Hz);

4,89 (1H, doublet, J=7,3 Hz);

lower than the 5.37 (2H, multiplet);

5,97 (1H, broadened. singlet);

? 7.04 baby mortality (1H, singlet);

9,80 (1H, broadened. singlet);

for 10.68 (1H, broadened. the singlet).

12) Range13C-nuclear magnetic resonance;

(mil.D. ve internal standard tetramethylsilane shown below:

174,3 (singlet), 170,4 (singlet), 168.6 (a singlet),

owed 161.1 (singlet), 137,9 (singlet), 135,7 (doublet),

135,1 (singlet), 129,8 (doublet), 116,3 (doublet),

115, 8mm (singlet), 113,7 (doublet), 77,6 (doublet),

74,4 (doublet), 72,1 (doublet), 71,8 (doublet),

66,0 (triplet), and 65.7 (doublet), 64,9 (triplet),

45,3 (doublet) and 43.9 (doublet, 36,6 (triplet),

33,4 (triplet), 30,1 (triplet), 30,0 (triplet),

29,7 (triplet), 27,0 (triplet), 26,7 (triplet),

20,3 (Quartet), and 16.6 (Quartet), and 16.3 (Quartet).

13) solubility:

Soluble in alcohols, such as methanol, ethanol, propanol and butanol; and soluble in dimethylsulfoxide, dimethylformamide, chloroform, ethyl acetate, acetone and ethyl ether; insoluble in hexane and water.

14) Color reactions:

Positive in sulfuric acid, iodine and potassium permanganate.

15) Thin layer chromatography:

The Rf-Value: 0,57.

Adsorbing agent Silica gel (Merek A Co.In.c. senior 5715).

Showing solvent: methylene chloride methanol 85:15 by volume.

Test example 1.

Antibacterial activity thimerisol.

Was determined minimum inhibitory concentration (MIC) thimerisol, expressed as mg/ml against gram-positive and Chemical Co. Ltd).

The results are given in table. 2.

Test example 2.

Antimycoplasma activity thimerisol.

Using the same method as in test example 1, was tested activity thimerisol in combating various strains of Mycoplasma. The results are given in table. 3.

The inoculate: 0,005 ml of 105CFU/ml.

Environment for testing:

M. Bovis and M. gallisepticum chanock medium [prepared as described in P. N. A. S. 48, 41-49 (1962) and supplemented with 20% horse serum]

M. Synovial Frey nutrient medium [prepared as described in Am.J. Vet.Res. 29, 2163-2171 (1968) and supplemented with 12% swine serum]

M. hyosinovial. Slimy secret LO*agar medium (supplemented with 15% horse serum).

The cultivation conditions: 37oC, 5 days, poor aeration (method BBL gas [cultivation of the air generator CO2frpm Becton Dickinson Microbiology System, Cockeyville, MD 2103 USA]

*PPLO (microorganism, such pleuropneumoniae).

PPLO broth without CV (Difco) 21 grams

Bacteriological mucous secretion (Difco) 5 g

Distilled water 800 ml

Agar noble (Difco) 12 grams

Horse serum 150 ml

25% fresh yeast extract 50 MLM

3. The method of producing antibiotic thimerisol formula

< / BR>
namely, that cultivated strain Alteromonas rava SANK 73390 and produce the target product from the culture fluid.


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FIELD: medicine.

SUBSTANCE: method involves cultivation of strain Streptoalloteichus cremeus subsp. tobramycini VKPM S-1084, producer of apramicyne. Emanation of apramicyne from culture fluid with sorption on carboxylic cationites followed by chromatographic purification with two-column method by stepwise elution with solutions (0.19-0.22)n of ammonia. The resulting eluate is concentrated in vacuum and precipitated with excess of acetone apramicyne in the form of sulfate.

EFFECT: increased output of apramicyne.

2 cl, 1 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: Amycolatopsis fructiferi subsp. ristomycini 8765 strain comes from mutant transformation of Proactinomyces fructiferi var. ristomycini 5339 strain properties as a result of consequent three-step actions on the stock strain by methylnitrosoguanidine, ultraviolet and gamma-radiation of 60Co with following selection of mutated strain possessing increased antibiotic activity. Obtained strain differs from already known one with growth form on diagnostic agar media, carbon sources assimilation rate (saccharose, starch, rhamnose, and mannose) chromogenic ability and antibiotic activity which is 5 times higher of Proactinomyces fructiferi var. ristomycini 5339 strain activity and is equal to 2000 mcg/ml. Strain is deposited to All Russian Collection of Industrial Microorganisms, collection # S1085.

EFFECT: invention can be used in medical industry for antibacterial antibiotic ristomycin manufacture; ristomycin is effective in treatment of infections induced by gram-positive microorganisms.

1 ex

Compound ws727713 // 2407785

FIELD: medicine.

SUBSTANCE: compound WS727713 is prepared by cultivation in a culture medium of Pseudonocardia FERM BP-7570 ray fungus strain producing such compound, and recovered from a culture fluid. The compound WS727713 is used as a melanocortin receptor modulator and applied for melanocortin receptor modulation while preparing a drug or a beauty preparation and preparing a pharmaceutical composition. Also, there are presented a pharmaceutical composition containing the compound WS727713, a method involving the introduction of specified compound in a human or animal body, and its application for treatment or prevention of the diseases responsive to melanocortin receptor regulation.

EFFECT: group of inventions exhibits the activity of a melanocortin receptor modulator.

12 cl, 3 dwg, 4 tbl, 1 ex