Method of delta4-3-ketosteroids 11 beta-hydroxylation

FIELD: medicine.

SUBSTANCE: invention refers to a method of Δ4-3-ketosteroids 11 β-hydroxylation by a biomass Curvularia lunata strain mycelium, RNCIM No. F-988. For the transformation, Curvularia lunata strain mycelium, RNCIM No. F-988 not older than 30 h and washed of nutrient medium is used. Mycelium is taken in such amount that the relation of the biomass to the transformed steroid makes 1.5-2.5:1. The transformation is performed in a buffer solution, and a steroid substratum is added as a microcrystal suspension, or as a water-soluble methyl - β-cyclodextrine complex with steroid related thereto as 1:1-0.6:1 (mol/mol). The yield of 11β-hydroxyderivatives is 50-80 %.

EFFECT: offered invention allows for higher selectivity of 11β-hydroxylation process, concentration of the transformed steroid substratum up to 20 g/l and reduced reaction period to 24-50 h.

1 tbl, 10 ex, 2 cl

 

The invention relates to the field of biotechnology, medicinal chemistry, Microbiology, and can be used in the pharmaceutical industry for the production of biologically active steroid compounds containing 11β-hydroxy-group, which is responsible for the presence in the molecule anti-inflammatory activity.

11β-hydroxylation of steroids is considered as one of the most important reactions in a series of steroid biotransformation, because it is used to retrieve a glucocorticoid anti-inflammatory and anti-allergic effects, such as hydrocortisone, prednisolone, and fluorinated corticosteroids (triamcinolone, dexamethasone, etc) on an industrial scale [1].

The maximum number of ways 11β-hydroxylation Δ4-3-steroids are ways to get hydrocortisone (C21About5H30, cortisol, a substance "F" Kendall, pregn-4-ene-11β,17α,21-triol-3,20-dione) is representative of an important class of drugs steroid [2-25]. As a self-medication/hydrocortisone is used to treat arthritis, bursitis, inflammatory and allergic skin diseases, etc. He also serves as the primary substrate in the synthesis of other corticoids - cortisone, prednisolone, prednisone, methylprednisolone [2]. Even 11β-hydroxylation need to receive dexamethasone scheme through 16α-methylcatechol.

Are also of interest as a compound of high physiological activity and other 11β-hydroxy Δ4-3-ketosteroids [22-28]. 11β-Hydroxysteroid get the introduction of the 11-hydroxyl group in the steroid molecule with the help of microorganisms - phycomycetes. However, unlike the 11α-hydroxylation, which carry out more than 300 species of microorganisms, the ability to accumulate 11β-hydroxysteroid with a satisfactory output, but in a mixture of 11α-hydroxyprogesterone inherent in a limited number of species of fungi Absidia orchidis, Cunninghamella blakesleeana, C.elegans, Cochliobolus lunatus[2, 6, 13, 16, 20, 28]. And only strains of Curvularia lunata (collections - ATS, IFO, NRRL, BKM and others) of the 35 known species widespread in nature, mold fungus of the genus Curvularia spend hydroxylation Δ4-3-ketosteroids mainly in the 11β-position and serve as a biocatalyst to get hydrocortisone [2-27]. Dignity species .lunata is the ability to 11β-hydroxylation as a 21-hydroxysteroids and 21-metalloceramic steroids[2, 3, 27].

Despite the very large number of studies performed with different strains of fungi, mainly with .lunata, the process 11β-hydroxylation remains the least manageable.

Found that 11β-hydroxylation using .lunata flows only in aerated conditions is, in the process of mycelium growth on nutrient medium and in its absence, i.e. using washed from the medium and suspended in the aqueous saline biomass. The transformation in the absence of the nutrient medium was performed using free or immobilized mycelium .lunata [5-10,13]. Water-insoluble steroid substrates were introduced into the reaction medium in the form of a solution in mixing with water, an organic solvent (ethanol, methanol, dimethylformamide, etc. or in the form of fine powder (MP) with a particle size of 10-15 µm [2]. However, due to the toxicity of the used solvents for fungi-transformers, the load of steroids in the reaction medium did not exceed 2 g/L.

There are ways of using steroid substrate cartaxana and its acetate order to increase their solubility in aqueous medium in the form of a complex with β-cyclodextrin, however, the solubility of the β-cyclodextrin does not allow you to raise the load steroid above 4 g/l [10, 19, 20]. The problem of increasing the solubility and availability of steroids for biocatalyst is not resolved in the works with protoplasts .lunata and with immobilized mycelium[5, 9, 13, 14].

There are several ways of introducing 11β-hydroxy-group with a growing culture .lunata. For example, various 17α-esters cartaxana and 17α-acetoxypiperidine used in the IDA MP in the amount of 0.25-2 g/l and transformed within 12-48 h [22]. 21-Acetate cortexone transformed in the hydrocortisone for 24 h growing crops .lunata KA-91 and .lunata CL-114 when the load of the substrate 1 g/l [19,20]. A famous example of 11β-hydroxylation 21-acetate cortexone cultures Cunninghamella blakesleeana ADS a and Cunninghamella echinulata F-1307943 that formed acetate hydrocortisone for 24 h under a load of acetate cortexone 0.15 g/l, dissolved in ethanol [21]. The method of introduction of 11β-hydroxy-group in the acetate norethisterone with a load of 1.25 g/l, dissolved in dimethylformamide. Transformation using .lunata ADS 12017 proceeded for 24 h [23].

In contrast to the above methods of transformation of steroid substrates with loads not exceeding 2 g/l, the process of introduction of 11β-hydroxy-group culture .lunata NRRL 2380 17-acetate 6α-methylpentadiene carried out when the load of the substrate 10 g/l, which was introduced into the nutrient medium in the form of MP and transformed 8-11 days [24].

However, for industrial applications the preferred ways 11β-hydroxylation, based on the use of biomass .lunata, suspended in buffer or saline solution. In this way received 16,17-acetonide hydrocortisone in the fermenter with a working volume of 500 liters of Substrate - 16,17-acetonide cortexone in the form of a solution in dimethylformamide were introduced into a suspension of mycelium prepared on distrurbance the water. When the load steroid 1 g/l transformation ended for 6-10 h [25]. Using mycelium .lunata F/70, suspended in phosphate buffer were obtained 11β-hydroxy-derived 16α-methyl-17α,21-dihydroxy-pregn-4-EN-3-one (a key compound in the synthesis of dexamethasone) when the load of the substrate is 0.2 g/l [26].

The disadvantages of the described methods 11β-hydroxylation are either low content transformable substrate, or a long period full of bioconversion. The disadvantages include the education side hydroxysteroids, which can reach up to five and that complicates the selection of the target product [2].

The known method according to which the transformable substrate used in the form of MP, which increased the content cortexone in the environment up to 10 g/l [7, 8]. However, the period of transformation when a load of the substrate carried out in phosphate buffer using free and immobilized mycelium .lunata VKM F-644, amounted to 168 hours Conversion cortexone in the hydrocortisone was performed using mycelium aged 48-60 h, i.e. washed from the nutrient medium in the stationary growth phase. Hydroxylation was performed with the same amount of mycelium to the ratio of the biomass to the steroid was 1:1 (based on dry biomass), and the process ended with the formation of hydrocort the area and two side hydroxysteroids.

We can assume that the number of mycelium was insufficient for the rapid conversion of 10 g/l cortexone. In this long process, conducted by the culture in hunger, inevitably it was age-related degradation, the result could be a decrease in selectivity and steroidgidroksiliruyushchei activity of the biocatalyst and the degradation products of transformation. Importance was also the age of the mycelium, which served as a biocatalyst. From literature it is known that the most high hydroxylase activity observed in mycelium in the logarithmic growth phase [2, 8, 12]. During aging culture in the reaction medium can receive the by-products of the transformation of steroid molecules and products of lysis of the mycelium, which adversely affect the yield of the target product and complicate his selection. In addition, since the water-insoluble steroids adsorbed on mycelium, the high content in the crystals cortexone can be a barrier to adequate access of oxygen to the biocatalyst. This hypothesis is confirmed by the data working with .lunata CL-114, whereby the output of hydrocortisone increased when the fractional flow of the substrate [20], and the results of the 11β-hydroxylation cortexone in the form of a water-soluble complex with β-cyclodextrin, the output of hydrocortisone which was the more than cortexone as MP [10, 20].

The aim of the invention is to devise effective way 11β-hydroxylation Δ4-3-ketosteroids with a new strain of Curvularia lunata VKPM F-988 with a token resistance to the antibiotic geneticin "G-418".

For the method is characterized by high speed and selectivity of the hydroxylation process in combination with the full conversion of the steroid substrate when the load is high. When the load transformable substrate to 20 g/l transformation occurs without degradation of steroids for a period of not more than 50 hours

The essence of the method lies in the fact that for 11β-hydroxylation use mycelium .lunata VKPM F-988, washed from the nutrient medium in the age of no more than 30 h, and the mycelium for the reaction to take in such quantity that the ratio of the density of the biomass of the fungus (dry weight) to the weight of the convertible steroid was 1.5 to 2.5:1.

Convertible steroids contribute to the reaction medium or in the form of microcrystals, MP, or soluble in the aqueous environment of the complex with chemically modified β-cyclodextrin, for example with methyl-β-cyclodextrin (MCD). It is known that chemically modified cyclodextrins increase the efficiency of bacterial processes 1,2-dehydrogenation of steroids and removal of the side chain of sterols [29]. However, transformations, Catala is projected mushrooms, the mechanism which differs from bacterial, chemically modified cyclodextrins were not applied.

Transformation of steroids in the form of a complex with WDC with .lunata VKPM F-988 proceeded in 2 times faster compared to the microcrystalline substrate. In addition, in this case facilitated the isolation of pure hydroxy, since the substrate and the reaction products will not accumulate on the mycelium (table), whereas the transformation of the substrate in the form of MP on mycelium contained almost half of the total number of steroids, which was extracted with intracellular substance.

This invention is illustrated by the following examples, the output of 11β-hydroxycodone (Y) which is calculated by the formula: Y=100×P/(S×(Qp/Qs), where P is the weight allocated crystalline product, S is the number taken in a reaction substrate, Qpthe molecular weight of the product, Qsthe molecular weight of the substrate.

Table
Transformation cortexone using mycelium .lunata VKPM F-988
Load cartaxana, g/lThe ratio of the steroid/methylcyclo-dextrin, mol/molTime transfer-mA is AI, hThe total content of the products of transformation, %The output of the crystalline hydrocortisone, %
In the aqueous phaseIn mycelium
10.0-4648.842.643.2
10.01/52291.5traces58.0
15.01/52496traces56.4
20.01/54898traces55.0
20.01/35076.610.552.0

Example 1. Transformation cortexone when a load of 10 g/L.

A spore suspension of the strain of Curvularia lunata VKPM F-988 grown at 28°C for 120-140 h h is beveled agar of the following composition (g/l): glucose - 4,0, yeast extract - 4,0, malt extract - 10, agar-agar - 25,0, distilled water, pH of 6.8, transferred into liquid medium in a conical flask with a volume of 750 ml with the medium of the following composition (g/l): sucrose - 60,0, yeast autolysate - 5,5, NaNO3- 3,5, NH4H2PO4- 6,5, K2HPO4- 2,0, KCl And 0.5, MgSO4to 1.0, pH 6.0-6.1, and grow the mycelium in aerobic conditions at a speed of mixing 220-250 rpm and a temperature of 28°C until the end stage of logarithmic growth. Received the seed material is transferred into a fresh medium of the same composition and spend growing in similar conditions. After 24 hours of growth obtained mycelium is filtered off, washed with distilled water, separated from the water, weighed and transferred in an amount corresponding weight 10-11 g dry biomass, 0.75 l of 0.07 M phosphate buffer, pH 6.0-6.1, containing 7.5 g of micronized steroid substrate with 95% of the basic substance - cortexone (7,12 g in 100%terms). The resulting suspension is distributed in 12 katalozhnyh Kolb and hold the transformation for 46 h (until the disappearance of the original substrate) in aerobic conditions, at a speed of mixing 220-250 rpm and a temperature of 28°C.

The mycelium is separated from the aqueous phase and extracted twice with ethyl acetate. The aqueous phase is shaken out three times with ethyl acetate, the extract is evaporated, dry OST the TCI weigh and determine the contents of steroid products using HPLC, then the weighted residues combine. Get to 6.75 g of a technical product, which is treated under stirring with 30 ml of hexane. Hexane is separated and the residue is dissolved in 60 ml of a mixture of CHCl3:Meon (7,5:1). To this solution dropwise with stirring, add 400 ml of water. The precipitate is filtered off, washed with a small amount l3, dried at 60°C. Obtain 3.2 g of crystalline hydrocortisone with TPL 211-214°C, identical to the standard sample (IR, PMR-spectra). The output of 43.2%. From chloroform Queen cells secrete 2.2 g (31%) of a mixture of 14α-hydroxy - 11β, 14α-dihydroxyquinoxaline (TLC and HPLC analysis).

Example 2. Transformation cortexone when a load of 10 g/l in the form of a complex with WDC.

Wet mycelium in the amount corresponding to the weight of 12-13 g of dry biomass obtained in example 1, make 1.0 l phosphate buffer containing 10 g cortesana (9.5 g 100%basis) and methyl-β-cyclodextrin (MCD) in respect of the steroid/WDC 1:1 (mol/mol). Incubated on a rocking chair 22 h under the conditions of example 1 including the Mycelium is separated from the aqueous phase extracted with ethyl acetate and determined by TLC absence of steroid products in the resulting extract. The aqueous phase is shaken out three times with ethyl acetate. After evaporation of the extract prepared 8,93 g technical product, which is treated analogously to example 1 and get 5,76 g (58%) Chris is alicebraga hydrocortisone with TPL 210-214°C. Of chloroform fraction obtain 2.1 g (21%) 14α-hydroxy-cortexone identical by TLC and HPLC standard sample.

Example 3.

Analogously to example 2 was performed transformation 3.75 g cortesana (3,56 g 100%basis) at a load of 15 g/l and was isolated and purified product transformation. The ratio of the biomass (in terms of dry weight of mycelium)/steroid was 2:1, the ratio of the steroid/WDC - 1/1 (mol/mol). The duration of the transformation Obtained 24 hours and 3.72 g of a technical product. Selected 2,01 g (56.4% of theory.) crystallic. hydrocortisone and 0.85 grams (23% of theory.) crystallic. 14α-hydroxy-cortexone.

Example 4.

Spent transformation cortexone with WDC at a load of 20 g/L.

a) Analogously to example 2 was performed transformation were isolated and purified product transformation. The amount loaded on the transformation of the substrate was 7 g (6,65 in 100%terms). The ratio of biomass/steroid was 2.5:1. Duration full conversion cortexone - 48 hours Obtained 6.0 g of a technical product. Selected 3,82 g (55% of theory.) crystallic. hydrocortisone and 1.5 g (21.5% of theoretical.) crystallic. 14α-hydroxycorticosterone.

b) same as (a) carried out the transformation of 5 g cartaxana and 4.75 g 100%basis), separation and purification of the reaction product. Used the ratio WDC/steroid 0,6/1 (mol/mol). The length of the transformer is then 50 PM Obtained 4.0 g of a technical product. Allocated 2.6 g (52% of theory.) crystallic. hydrocortisone and 0.95 g (19% of theory.) 14α-hydroxycorticosterone.

Example 5. Transformation 21-acetate cortexone when a load of 10 g/L.

Analogously to example 2 transformed 5 g 21-acetate cortesana (4.5 g in 100%terms) within 48 hours of the Selection were performed according to example 2. Received 3,36 g technical product. Selected 2.15 g (51%) of theory.) crystallic. hydrocortisone and 0.84 g of 14α-hydroxy-cortexone (21% of theory.).

Example 6. Obtain 16α-methylhydroquinone of the 21-acetate 16α-methyl cortexone.

The transformation of 3 g 21-acetate 16α-methyl-pregn-4-EN-17α,21-diol-3,20-dione in the form of a complex with WDC (with a ratio of the steroid/WDC 1/4) was performed at a load of steroid 10 g/l for 40 h using the ratio of the biomass to the steroid of 2.5:1. The mycelium does not contain transformation product, was separated from the aqueous phase. The aqueous phase was extracted with ethyl acetate. The extract was evaporated. To the residue was added an equal volume amount of chloroform, precipitated precipitate was filtered and dried at 65°C. Received 1.32 g technical 11β-hydroxypropyl (yield 50%). After crystallization, the product was identical to the sample 16α-methyl-hydrocortisone [26].

Example 7. Getting 11β-hydroxytestosterone of testosterone.

The transformation of 4 g of chemically pure testosterone was performed in the conditions of example 2 in t is the increase in the 24 h under a load of 5 g/L. The mycelium and the aqueous phase was extracted separately. Extracts containing data on TLC about the same amount of reaction product were combined, evaporated and obtained 3.8 g of a technical product, which was dissolved in 30 ml of methanol. To a methanol solution was added 5 ml of water. The resulting mixture was cooled to 5-7°C for 3 hours was Separated crystalline precipitate. Washed it cold methanol, dried and received 2,53 g (yield 60%) 11β-hydroxytestosterone with TPL 232-235°C. Lit-236.5°C [30].1N PMR (CDCl3, δ) of 1.01 (3H, s, H-18), OF 1.42 (3H, s, H-19), to 3.58 (1H, apt,3J=8,8 Hz, H-17), 4,36 (1H, m, J=9.4 Hz, H-11), THE 5.65 (1H, s, H-4).

Example 8. Getting 11β,17α-dihydroxyprogesterone.

The transformation of 2.4 g of chemically pure 17α-hydroxyprogesterone was performed in the conditions of example 2 for 24 h under a load of 5 g/L. Mycelium and the aqueous phase was extracted separately. After evaporation of the extract of the aqueous phase (TLC showed the absence of steroids on mycelium) was obtained 2.2 g of a technical product, which was washed 1-1,2 ml of acetone. Obtained 2.0 g (80%) of crystalline 11β,17,α-dihydroxyprogesterone, TPL 223-225°C. Lit-228°C. [31].1N PMR (CDC13, δ): 1,0 (3H, s, H-18), 1,4 (3H, s, H-19), AND 2.27 (3H, s, H-21), of 2.72 (1H OSS, 17-one), OF 4.45(1H, m, H-11α), to 5.66 (1H, s, H-4).

Example 9. Getting 11β-hydroxy 20-hydroxy-methylpregna-1,4-Dien-3-one (GND).

Analogously to example 2 transformed 1.0 g AME is in a period of 24 hours at a load of 4 g/L. The aqueous phase was extracted with ethyl acetate (mycelium steroids were absent). After evaporation of the extract obtained 0.95 g of a mixture of hydroxy GMPD that I shared on preparative SiO2. Steroids were suirable a mixture of CHCl3/acetone 7:2. Selected 0.45 g (43%) 11β,20-dihydroxyethylene-1,4-Dien-3-one (1N PMR (CDCl3, δ): 0,86 (3H, s, H-18), OF 1.35 (3H, s, H-19), IS 4.15 (1H, q, H-11), 5.8 (1H, s, H-4), 6,03 (1H, d, H-2), from 7.24 (1H, d, H-1) and 0.26 g (25%) 7β,20α-dihydroxyethylene-1,2-Dien-3-one.1N PMR (CDCl3, δ): 1,02 (3H, H-18), 1,24 (3H, s, H-19)and 4.2 (1H, t, J=15,5 Hz, H-7), 6.1 (1H, d, H-4), 6.2 (1H, d, H-2), 7.05 (1H, m, H-1).

Example 10. Getting 11β-hydroxy progesterone.

Analogously to example 2 transformed 1.0 g of progesterone for 25 h under a load of 2 g/L. Aqueous phase was extracted with ethyl acetate (mycelium steroids were absent). After evaporation of the extract obtained 0.8 g of a crystalline precipitate consisting of a mixture of 2 hydroxy progesterone in the ratio of 1:1, identified as 11β-hydroxyprogesterone (1N PMR (CDCl3, δ): 0,99 (3H, s, H-18), WAS 1.43 (3H, s, H-19), AND 2.1 (3H, s, H-21), 3.16 (1H, m, H-17), THE 4.29 (1H, m, H-11α), 5.66 (1H, s, H-4) and 6α-hydroxyprogesterone (1N PMR (CDCl3, δ): 0,73 (3H, s, H-18), TO 1.19 (3H, s, H-19), AND 2.1 (3H, s, H-21), AND 3.16 (1H, m, H-17), 4.39 (1H, m, H-6β), 5,79 (1H, s, H-4).

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1. The way 11β-hydroxylation Δ4-3-ketosteroids with the fungus Curvularia lunata, characterized in that for carrying out the process using biomass of mycelium of the strain of Curvularia lunata VKPM F-988, and transformed steroid contribute in the form of a suspension of microcrystals or preferably in the form of a complex with methyl-β-cyclodextrin at a ratio of methylcyclohexene-steroid 1:1-0,6:1 (mol/mol).

2. The method according to claim 1, characterized in that the age of the mycelium Curvularia lunata does not exceed 30 h, and the ratio of the biomass of mycelium transformed to the steroid is 1.5-2.5:1.



 

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2 dwg, 3 ex

FIELD: medicine.

SUBSTANCE: method implies DNA recovery of an investigated strain, polymerase chain reaction (PCR) with oligonucleotide primers presented in the patent claim, and determination of a nucleotade sequences of amplified rjg, gmhD and orf2 gene fragments and their comparison to similar genes of comma bacilli stains of the others serogroups. The genetic relationship of comma bacilli stains of various origins is determined by a number of individual nucleotide substitutions in rjg, gmhD and orf2 genes flanking a cluster of O-antigen biosynthesis, in comparison similar genes of comma bacilli stains of the others serogroups presented in GenBank database.

EFFECT: use of the invention allows for rapid, effective and reliable determination of relationship of comma bacilli stains of various origins with the other comma bacilli, including with virulent and epidemically significant vibrios of O1 and O139 serogroups.

2 tbl, 8 ex

FIELD: medicine.

SUBSTANCE: invention is intended for a quantitative estimation of adhesive properties of staphylococci recovered from a human and environment objects, with its bacteriological identification and for characterisation of its virulent properties in vitro by the adhesion to biopolymeric molecules. The staphylococcus culture being investigated is incubated in wells of a polystyrene plate with immobilised haemoproteins (muscle haemoglobin, haemoglobin) in a nutrient medium 199 for 1 hour; thereafter, nonadherent bacteria are removed by triple washing in the medium 199 with Twin-20, and a number of adhesive staphylococci is estimated by harvest cells and shown by optical density of the medium and by counting the cells sown on the dense nutrient medium from a series of tenfold cultivations, or by analysing the adhesive strains for the activity of bacterial enzymes.

EFFECT: good results reproducibility, high sensitivity of the method, ease of the reaction directly in wells of the polystyrene plate and possible express estimation of adhesive capacity of strains.

3 tbl, 3 ex

FIELD: food industry.

SUBSTANCE: method provides for depth cultivation of basidium fungi Pleurotus ostreatus on nutrient medium of the following composition (g/l): soya flour - 21, KH2PO4 - 3.5, MgSO4 - 0.4, milk whey - 200 ml, water - up to 1 litre. Mycelium is exposed to deep freezing with further defrosting. Then its disintegration is carried out. Food fibres are cleaned by alternating treatment with cold and hot water, mix of 1 M sodium carbonate or sodium hydroxide with ethanol in the ratio of 1:2 and 0.5 M solution of citric acid. Yield of food fibres makes 16-18% of dry biomass, content of chitin - 16-19%, glucan - 40-50% of dry mass of food fibres.

EFFECT: food fibres with high content of chitin.

4 ex

FIELD: chemistry; biochemistry.

SUBSTANCE: invention relates to biotechnology and can be used for synthesis of n-butyl alcohol, acetone and ethanol, as well as production of solvents. The Clostridium acetobutylicum VKM B-2512D strain is deposited in the Russian collection of microorganisms of the Institute of Biochemistry and Microbial physiology of the Russian Academy of Sciences and is a producer of n-butyl alcohol, acetone and ethanol.

EFFECT: invention increases output of n-butyl alcohol, acetone and ethanol and widens the raw material base for their production.

1 tbl, 1 ex

The invention relates to the field of biotechnology and can be used in pharmaceutical, microbiological and chemical industry for preparation of intermediates for the synthesis of steroid drugs
The invention relates to biotechnology and can be used in pharmaceutical, microbiological and chemical industry for preparation of intermediates for the synthesis of steroid drugs

FIELD: medicine, pharmaceutics.

SUBSTANCE: there are described compounds of the following structure or their salts: where A, B, R2, R4, R6, R7, R10, R16, R17α, R17β, Z, Y, X have the values specified in the description. Some of these compounds exhibiting tissue-specific antiandrogen activity and tissue-specific androgen activity can be applied for treating or reducing risk of the diseases associated with androgen stimulation loss.

EFFECT: preparation of the compounds which reducing probability of the androgen-dependent diseases, such as prostate cancer, benign prostate hyperplasia, polycystic ovary syndrome, acne, hirsutism, seborrhoea, etc.

41 cl, 176 ex, 4 tbl

Organic compounds // 2387664

FIELD: medicine.

SUBSTANCE: there are described compounds of formula in a free form or in the form of salt where R1 and R2 have values specified in the description of the application which are used for treating inflammatory conditions, first of all inflammatory or obstructive respiratory tract diseases. Besides the application describes the pharmaceutical compositions containing said compounds, and methods for preparing said compounds.

EFFECT: compounds exhibits improved efficiency.

5 cl, 8 ex

FIELD: chemistry.

SUBSTANCE: described is a 15β-substituted oestradio derivative having selective oestrogenic activity. The preferred compound is 7α-ethyl-15β-methyl-19-nor-17α-pregna-1,3,5(10)-trien-20-yne-3,17β-diol.

EFFECT: obtaining compounds which can be used in treating or preventing diseases or physiological conditions related to oestrogen receptors.

11 cl, 4 ex, 2 tbl

FIELD: medicine.

SUBSTANCE: invention relates to new 3.15-substituted estrone derivatives that are inhibitors of 17β-hydroxysteroid-dehydrogenase type 1 (17β-HSD1), to their salts, pharmaceutical compositions, containing specified compounds and to methods of such compound producing. Besides the invention refers to application in medicine of specified new 3,15-substituted estrone derivatives firstly to their application for treatment or prevention of steroid-dependent diseases or disorder such as steroid-dependent diseases or disorders treatment of which requires inhibition of 17β-hydroxysteroid dehydrogenase type 1 and/or reducing of endogenous 17β-estradiol concentration. The invention also relates to general application of selective inhibitors of 17β-hydroxysteroid-dehydrogenase type 1 that do not bind with estrogen receptor or display antipathic affinity to estrogen receptor.

EFFECT: possibility of application for treatment and prevention of benign gynaecologic diseases.

49 cl, 836 ex, 42 tbl

FIELD: chemistry.

SUBSTANCE: invention refers to synthesis of [18F]fluororganic compounds ensured by reaction of [18F]fluoride and relevant halogenide or sulphonate with alcoholic vehicle of formula 1 where R1, R2 and R3 represent hydrogen atom or C1-C18 alkyl.

EFFECT: possibility for mild process with low reaction time and high yield.

21 cl, 2 tbl, 27 ex

FIELD: technological processes.

SUBSTANCE: invention is related to automation of technological processes and may be used in automation of process of production of loose form of powdery choline chloride from its aqueous solution. In method that provides for use of crushed and fractionated dry sugar beet pulp as active adsorbent, its mixing with previously heated aqueous solution of choline chloride, and then drying in vibration dryer by superheated steam of atmospheric pressure, separation of spent superheated steam flow into the main one, sent to vibration dryer with creation of recirculation circuit, and additional one sent for reheating of choline chloride prior to its supply for mixing, the novelty is the fact that superheating of atmospheric pressure steam is done with heating steam, at that heating steam is produced by means of steam generator with electric heating elements, feed pump and safety valve, heating steam condensate produced in this process after superheating and condensate produced during heating of aqueous solution of choline chloride is taken to condensate collector, and then in mode of closed circuit is supplied in steam generator, at that flow rate of crushed and fractionated dry pulp is measured, as well as aqueous solution of choline chloride coming for mixing, flow rate and temperature of superheated steam upstream vibration dryer, choline chloride temperature before and after its heating, pressure of choline chloride after heating, temperature and humidity of mixture of crushed and fractionated dry pulp and aqueous solution of choline chloride prior to supply for drying, amplitude and frequency of oscillations in gas-distributing grid of vibration dryer, flow rate and humidity of powdery choline chloride after drying, level of condensate in steam generator and pressure of heating steam, at that flow arte of dry sugar beet pulp after fractionation is used to set flow rate of heated choline chloride coming for mixing, and flow rate and humidity of prepared mixture of crushed and fractionated dry pulp and aqueous solution of choline chloride prior to supply for drying, flow rate and humidity of powdery choline chloride after drying are used to determine amount of evaporated moisture in vibration dryer, which is used to establish flow rate of superheated steam in the main circuit of recirculation, and its temperature is established by current value of temperature of mixture of crushed and fractionated dry pulp and aqueous solution of choline chloride by setting of specified capacity of steam generator affecting power of electric heating elements, at that in case condensate level in steam generator falls below specified value, condensate is supplied from condensate collector, and when pressure of steam in steam generator reaches upper limit value, steam pressure is released through safety valve, if flow rate of mixture of crushed and fractionated dry pulp and aqueous solution of choline chloride deviates prior to supply for drying to the side of increase from specified value, at first frequency is increased, and then amplitude of oscillations in gas-distributing grid of vibration dryer, if flow rate of mixture of crushed and fractionated dry pulp and aqueous solution of choline chloride deviates to the side of reduction from specified value, at first frequency is reduced, and then amplitude of oscillations in gas-distributing grid of vibration dryer, current values of temperature and flow rate of choline chloride prior to heating are used to set flow rate of spent superheated steam in additional recirculation circuit, at that temperature of choline chloride after heating is used to set specified pressure of choline chloride at the inlet to mixer.

EFFECT: provides for increased quality of finished product, accuracy and reliability of control, increased yield of finished product, reduced specific heat and power inputs and prime cost of finished product.

1 dwg

FIELD: chemistry.

SUBSTANCE: invention concerns novel compounds of formula I: , where M is macrolide subunit of substructure II: , L is chain of substructure III: -X1-(CH2)m-Q-(CH2)n-X2-, D is steroid or non-steroid subunit derived from steroid or non-steroid NSAID medicines (nonsteroid anti-inflammatory drug) with anti-inflammatory effect; pharmaceutically acceptable salts and solvates of claimed compounds; methods and intermediary compounds for obtainment of claimed compounds.

EFFECT: improved therapeutic effect, application in inflammatory disease and state treatment for humans and animals.

37 cl, 18 ex

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