Steroid compounds, method for selective modification, pharmaceutical composition

FIELD: organic chemistry, steroids, medicine, pharmacy.

SUBSTANCE: invention relates to steroid compounds of the formula (1)

wherein --- means optional double bonds; R6 means hydrogen atom (H), =CH, -CH3 or -CH2-CH3; R7 means hydrogen atom (H), (C1-C4)-alkyl, (C2-C5)-alkenyl, or (C2-C5)-alkynyl; R11 means hydrogen atom (H), (C1-C4)-alkyl, (C2-C4)-alkenyl, (C2-C4)-alkynyl, (C1-C4)-alkylidene; E means 5-7-memberd ring formed with 16 and 17 carbon atoms at α,cis-position relatively to steroid structure and comprising possibly up to two double bonds. Compounds can be used in therapy and in methods for selective modification of activity of estrogen receptors.

EFFECT: improved method for modifying, valuable medicinal properties of compounds.

10 cl, 1 sch, 1 tbl, 1 ex

 

The present invention relates to steroid compounds with additional ring E, associated with the D ring of the steroid structure, possessing estrogenic activity.

There is a constant interest in new compounds with affinity for the estrogen receptors. This occurs as a result of opening two different receptor subtypes, denoted by ERα ERβ (see Mosselman et al., FEBS Letters 392 (1996) 49-53, as well as EP-A-0798378). Compounds that are selective with respect to these receptor subtypes may provide a more selective therapeutic effects associated estrogen receptors. Advantages, for example, can be obtained from the various distribution of receptor subtypes in human tissues. This allows for a treatment with fewer side effects associated with estrogen. Examples of therapeutic use of estrogen, which can be improved with the use of selective compounds are contraception, treatment of symptoms of menopause, osteoporosis and treatment estrogenzawisimy tumors.

In EP 0869132 described estrogenic steroid compounds having the formula (1):

where:

dashed links represent optional double bonds;

R6represents H, =CH2or-CH3or-CH 2-CH3;

R7represents H, C1-4-alkyl, C2-5alkenyl or2-5-quinil, where alkyl, Alchemilla or Alchemilla group may be substituted by 1-3 halogen atoms, independently selected from the group of fluorine atoms and chlorine;

R11represents H, C1-4-alkyl, C2-4alkenyl,2-4-quinil or C1-4-alkyliden, where alkyl, Alchemilla, Alchemilla or alkylidene group may be substituted by 1-3 halogen atoms, independently selected from the group of fluorine atoms and chlorine;

E represents together with the carbon atoms 16 and 17 of the steroidal structure four - semiline ring, and the specified ring is α in the CIS-position relative to the steroid structure optionally contains one to two vnutrirossijskikh relations and substituted REthat has a number of values.

Later in the EP 0869132 revealed that any alkyl, Alchemilla, Alchemilla and alkylidene group of steroid compounds represented by formula (1)may be razvetvlennoy or unbranched. If R6or R11attached to the steroid structure via a single bond, a substituted carbon atom of the steroid structure or linked to a hydrogen atom, or forms a double carbon-carbon bond. The connection may include various centers hir is lesti and can exist as enantiomers and diastereomers. Hydroxyl groups can be protected by such substituents as acyl or alkyl, forming prodrugs of the compounds represented by formula 1.

It is now established that steroid compound represented by formula 1 with symbols and signs, with the above values, characterized by the fact that REis a βa hydroxy-group or its prodrug, unexpectedly, has a higher selectivity for the estrogen receptor α in combination with greater efficiency estrogen α. This connection, hereinafter, referring to the connection according to the present invention, are usually not only very weakly active against the estrogen receptor βbut essentially represents a General antagonist of the estrogen receptor βthat exhibits very high selectivity with respect to estrogen receptor α and, moreover, allows for selective therapy based on the blockade of the estrogen receptor β. Compounds according to this invention include the above enantiomers and diastereomers included in the scope of the invention, and each of the individual (R)- and (S)-enantiomers, essentially free, i.e. contains less than 5%, preferably less than 2% and especially less than 1% other enantiomer is a, and mixtures of such enantiomers in any ratio, including racemic mixtures consisting mainly of equal amounts of two enantiomers.

The preferred embodiment of the invention is a steroid compound that is listed above and, in addition, characterized by the fact that R6represents H, R7is a branched or non-branched C1-3-alkyl, ring E represents a five - or six-membered ring contains no double bonds and containing a hydroxy-group in position 22 S-stereoconfiguration, in other words, this configuration is a β in relation to steroid structure according to the notation adopted in the stereochemistry of steroid compounds.

Most preferred is a compound represented by formula 2, which is (7α,16β,17α,22S) -7-propyl-16, 24-cyclo-19, 21-Generala-1,3,5(10)-triene-3,17,22-triol, having a code cipher Org 41621:

Compounds according to the present invention therefore allows for a more directed, in other words a selective modification of the activity of estrogen receptors α or β in the body.

The prodrug is defined as the connection preobrazovyvayutsya in the body of the recipient in the compound represented by formula 1, the de R Eis a β-hydroxyl group. It should be noted that the hydroxy-group in position 3, 17 and the ring E, for example, can be converted to ethers (alkyl*hydroxy) or esters, such as acyl*hydroxy, phosphate, sulfate, sulfonate or aromatic carboxylate, with the carbon chain length groups marked with an asterisk (*)are not considered to be severely limited. Acyl group derived from a linear or branched alkane*, and aromatic carboxylate will usually contain a phenyl, pyridinyl or pyrimidyl. The length of the alkyl and acyl groups selected depending on the desired properties of the prodrugs, the more long chain prodrugs containing, for example, laureline or karolneu chain, are more appropriate for drug slow release and Deposit products. It is known that such alternates spontaneously hydrolyzed or hydrolyzed enzymatically to free hydroxyl groups, the replacement serodino the connection structure. Such prodrugs will be comparable biological activity of those compounds in which they are converted in the body of the recipient. Active connection, which converts the prodrug, called the parent, the source connection. The onset of action and duration of action as well as the distribution is giving prodrugs in the body may differ from the similar properties of the original connection.

As for medication, and physiological, medical and pharmacological experiments such selective compounds presented here are extremely necessary. One aspect of the invention is that the connection according to the invention can be used for treatment by administering compounds to a recipient, being a person or animal, preferably mammals. The different distribution of receptors α and β in various tissues of the body provides a target for more selective interference with the functioning of different tissues. It is known that receptors dependent, estrogen receptors α preferably expressed in vaginal tissue and liver tissue, whereas estrogen receptors β preferably expressed in prostate tissue, the epithelial cell layer of the urinary bladder of rats, endothelial, smooth muscle cells of blood vessels and certain areas of the brain such as the basal forebrain, cerebral cortex (neocortex and hippocampus. Fabrics, which are both types of receptors are, for example, the tissue of the pituitary gland, hypothalamus, thymus, uterus, ovary and bone.

Profile of affinity to the estrogen receptors of the compounds according to the present invention makes their PR is good as improved estrogen compounds or antiestrogenic compounds with reduced side effects, associated with estrogen. Thus, the invention represents a method for selective modification of the activity of estrogen receptors by implementing a contact interaction of the compounds according to the invention with estrogen receptors. This method may be the treatment of a human or animal, but it can also be a non-medical way. The latter may represent an experimental way, such as estimating the selectivity of compounds or laboratory method in vitro information on estrogen receptors or compounds that interact with them. Preferably, these compounds can be used for selective in relation to estrogen receptors α and β contraception, experimental or medical therapy, for example, for the treatment or prevention of disorders associated with estrogen receptors, symptoms of menopause, osteoporosis, cardiovascular disorders, modulate pituitary hormonal regulation, benign prostate hypertrophy, control estrogenzawisimy tumors, malignant tumors of the colon, endometriosis or disorders of the Central nervous system. An important common characteristic of these selective methods and treatment methods is that they include the implementation in which aimogasta compounds according to the invention with estrogen receptors.

The invention also relates to the use of compounds according to the invention for obtaining a medicinal product for selective therapy involving estrogen receptors, and drug for the treatment of disorders associated with estrogen receptor αcomprising the administration to a patient compounds according to the invention (in an acceptable pharmaceutical dosage form). Whereas the antagonistic effect of the compounds according to the invention in relation to the estrogen receptor β, the invention also provides for obtaining a medicinal product for the treatment of disorders associated with estrogen receptor βcomprising the administration to a patient compounds according to the invention (in an acceptable pharmaceutical dosage form).

In addition, the invention relates to the use of compounds according to the invention for obtaining a medicinal product with contraceptive activity. Thus, the invention also relates to the medical indication of contraception, i.e. the method comprising administration to the patient, which is the woman, or female animal type of progestogen and estrogen in accordance with the conventional scheme in this area, while estrogen is a compound according to this invention (in an acceptable formats ticheskoj dosage form).

Finally, the invention relates to the use of compounds according to the invention for obtaining a medicinal product with selective estrogenic activity, such drug is mostly applicable in the field of HRT (hormone replacement therapy), characterized by softening of the complications of menopause, in particular protivoseborainey activity.

Metered quantities of these compounds are within the usual procedure for compounds related to estrogen, for example, from 0.01 to 100 mg per dose.

This purpose can be obtained from single dose containing compound according to the invention in quantities of the same order as above therapeutic doses. Therefore, the present invention also relates to pharmaceutical compositions containing the compound according to the invention in a mixture with one or more pharmaceutically acceptable auxiliary substances.

Compounds according to the invention can be obtained using various methods known in the field of organic chemistry in General and especially in the field of steroid chemistry. See, for example: J. Fried and J. A. Edwards, Organic Reactions in Steroid Chemistry", Volumes I and II, Van Nostrand Reinhold Company, New York, 1972; and C. Djerassi, "Steroid Reactions ", Holden-Day, Inc., San Francisco, 1963.

For the synthesis of compounds according to the invention was synthesized steroid joint is additional 16,17-analyoung ring. These methods are described in EP 0869132. For compounds according to the present invention an additional hydroxyl functional group must be entered in annulirovano ring. To this end it is convenient for the reaction anaerobia so that the procedure of synthesis was located appropriately double bond, for example, using the well-known exchange reactions of olefins, what is it used for metal catalysts, obtained, for example, ruthenium, molybdenum or tungsten, for circuit 16α, 17α-binnensysteme fragment in unsaturated annulirovano 5 - or 6-membered ring. Synthesized in a similar manner olefinic ring first stereospecific epoxidized. This reaction can generally be carried out using such agents as percolate (preferably in buferiruemoi environment), or catalytic systems using metal complexes in the presence of oxidizing agents (such as hydrogen peroxide or tert-butylhydroperoxide). In this case, usually, good results are obtained by applying perbenzoic acid buffer solution of sodium bicarbonate. The epoxides can be almost selectively restorative disclosed for a particular position using hydride reagents to the desired beta-alcohols. Hydroxy-group can also b shall be readily synthesized using techniques hydroporinae/oxidation. In the case of obtaining α-hydroxycodone application of the Mitsunobu reaction easily leads to intended β-alcohols. Hydroxycodone can be converted into prodrugs, such as alkyl esters, acyl esters, carbonates, sulfonates or phosphates, through reaction with a suitable alkylhalogenide or acid chloride of the acid, if desired.

Pharmaceutical composition containing one or more compounds according to the invention, can be obtained by combining with pharmaceutically acceptable excipients or without the indicated combination, as described in the standard reference, Gennaro et al., Remmington''s Pharmaceutical Sciences, (18th ed.. Mack publishing Company, 1990, see especially Part 8: Pharmaceutical Preparations and Their Manufacture). A mixture of one or more compounds according to the invention and one or more pharmaceutically acceptable excipients may be compressed into solid standard dosage forms, such as pills or tablets, or can be put into capsules or suppositories. By means of pharmaceutically acceptable liquids compounds can also be used as an injectable in the form of a solution, suspension, emulsion or spray, for example a nasal spray. Compounds according to the invention can also be incorporated into the implant, IUD, patch, gel, and Liu is Oh another drug for long-term (delayed) release. To obtain a standard dosage forms, such as tablets, it is assumed the use of conventional additives such as fillers or carriers, colorants, polymeric binders and the like. Basically can use any pharmaceutically acceptable additive, which does not affect the function of the active compounds. Suitable carrier materials, together with which can be entered compositions include lactose, starch, cellulose derivatives and the like or mixtures thereof, used in acceptable quantities.

EXAMPLES

The way of synthesis used in the examples illustrated in schemes I and II. Rooms used to refer to compounds characterized by the structural formula on the data schemas.

Connection 2

To a solution of the 5.65 ml vinyltrimethylsilane in 50 ml of anhydrous THF ml) was added at -50°With a drop of 12 ml of 1.6 M BuLi in hexane. After stirring for an additional 0.5 h solution was added 6.2 g of 16α-allyl, 7α-telectron-3-O-methyl ether (1) in 20 ml of anhydrous THF at -50°C. After stirring for an additional 1/2 h the mixture was poured into a feast upon. NH4Cl and extracted with ethyl acetate. Concentration of the organic phase followed by chromatography on silica gel gave 4,2 g 2 in the form of oil,

Rf,47 (toluene/ethyl acetate 95/5), for 1 Rf0,65. NMR (CDCl3) δ 5,80 (m, 1, CH-allyl), 6,07 (m, 1, CH vinyl), and 0.98 (s, CH3), of 0.93 (t, 3, ethyl), of 3.78 (s, 3, CH3).

Connection 3

To a solution of 4.2 g of 2 in 80 ml of methylene chloride was added 0.32 g of dichloride benzylideneacetophenone (catalyst exchange Grubbs). After stirring for 1 h was added an additional portion of 0.3 g of catalyst. After completion of the reaction (2 h) and the mixture was concentrated, and the residue was purified by column chromatography to obtain 3.5 g 3, Rf0,29 (heptane/ethyl acetate 8/2, 2 Rf0,55).

NMR (CDCl3) δ 5,72 (m, CH=), of 6.02 (m, 1,=), 0,99 (s, 3, CH3), to 0.89 (t, 3, CH3), of 3.77 (och3), to 0.92 (m, 3, CH3).

Connection 4

A mixture of 0.5 g of steroid compounds 3 and 0.5 g Panso3in 12 ml of methylene chloride was treated with 0.34 g of meta-Cl-perbenzoic acid. After stirring for several hours at room temperature the reaction mixture was diluted with water and treated with sodium thiosulfate solution for the destruction of the remaining peroxide. Organic matter was extracted with ethyl acetate and finally purified using chromatography to obtain 110 mg of the desired β-epoxide 4; Rf0,50 (toluene-acetone 9/1).

NMR (CDCl3) δ and 0.98 (t, 3, CH3), to 0.92 (t, 3, CH3), 3,65 + 3,70 (2xm, epoxide CH).

Connection 5

A solution of 80 mg steroide what about connect 4 in 3 ml of THF was heated under reflux with 10 mg LiAlH 4. After 2 h starting material disappeared. The mixture was suppressed by addition of 30 μl of the feast upon. solution of Na2SO4and 0.20 g of Na2SO4was stirred for 15 min and filtered through celite. The filtrate was concentrated and the residue was passed through a short column with silica gel to obtain 55 mg 5; Rfof 0.24 (toluene-acetone 9/1); NMR (CDCl3) δ Android 4.04 (m, 1, SNON), of 3.78 (s, 3, och3), 2,85 (m, 2, CH2when C6), to 0.92 (s, 3, CH3).

Connection 6

To a solution of Acantilado sodium (derived from 0.7 ml of ethanthiol and 0.27 g of 60% dispersion of NaH) in 9 ml of DMF was added 120 mg of steroid compounds 5. The mixture was heated under reflux for 3 h Then the reaction mixture was poured into water and was extracted with ethyl acetate. Conducted chromatography of organic matter to obtain 80 mg of 6, the melting point 224-226; Rf0,30 (toluene-acetone 8/2); NMR (CDCl3) δ 4,00 (m, 1, SNON), 7,12 (ar H1), 6,20 (ar H2), 6,54 (ar H4).

Connection 7

To a solution of 2.7 g of steroid compounds 3 in 20 ml of methylene chloride and 3 ml of pyridine was added at 0°With 1.9 ml of tributyltinchloride. After stirring for 1/2 h, the reaction mixture was poured into water and was extracted with ethyl acetate to obtain 3.3 grams essentially purified similarverizon 7, Rf0,8 (heptane-acetone 8/2); NMR (CDCl3) δ of 0.03 (s, 9, TMS), 5,68, 5,91 (Hz, CH olefin).

Compound 8

A solution of 3.2 g of 7 in 25 ml of espagnole THF was treated at 0° With a solution of 2.2 ml of borane-dimethylsulfide complex in 20 ml of THF. The reaction mixture was further stirred for 2 h at 45°C. the Excess reagent was destroyed by careful addition of 4.5 ml of abs. ethanol, followed by addition of 11 ml of 2 N. NaOH and 7.7 ml of 30% hydrogen peroxide. The reaction mixture was stirred over night, diluted with water and extracted with ethyl acetate. The thus obtained crude product was purified by column chromatography and obtained 1.7 g of the desired α-alcohol 8. Rf0,36 (heptane-acetone 8/2); NMR (CDCl3) δ was 4.42 (m, 1, SNON), of 0.12 (s, 9, TMS), of 0.79 (s, 3, CH3).

Connection 9

A solution of 1.6 g of 8 and 1.3 g of triphenylphosphine in 60 ml of toluene was treated with 0.84 g of para-nitrobenzoic acid and 0.8 ml of diethylazodicarboxylate at 0°C. After stirring for 1 h the reaction was completed. The mixture was poured into a feast upon. an aqueous solution Panso3and were extracted with ethyl acetate. After chromatographic purification was obtained 2.9 g β-nitrobenzoate, Rf0,64 (heptane-acetone 8/2); NMR (CDCl3) δ of 5.34 (m, 1, CHOC(O)Ar)of 0.91 (s, 3, CH3), of 0.95 (t, 3, CH3), of 3.80 (s, 3, och3). This substance was dissolved in a mixture of 40 ml of THF-methanol (1/1 by volume) and was treated with 4 ml solution of 2 N. NaOH. After stirring for 15 min the reaction mixture was introduced into water and the product was extracted with ethyl acetate. After making the Chania through a short column with silica gel was obtained 1.3 g β -alcohol 9; Rf0,64 (heptane-acetone 8/2); NMR (CDCl3) δ or 4.31 (m, 1, SNON).

Connection 10

A solution of 1.3 g of 9 in 30 ml of acetone was treated with 2 ml of 2 N. Hcl. After 1 h the mixture was neutralized by adding saturated Panso3and concentrated. The residue was diluted with water and extracted with ethyl acetate to obtain 1.0 g 10, Rf0,10 (heptane-acetone 8/2); NMR (CDCl3) δ of 0.90 (t, 3, CH3), of 0.95 (s, 3, CH3), 4,48 (m, 1, SNON).

Connection 11

To a solution of Acantilado sodium (derived from 0.7 ml of ethanthiol and 0.3 g of 60% dispersion of NaH) in 9 ml of DMF was added 120 mg of steroid compounds 10. The mixture was heated under reflux for 3 hours Then the mixture was introduced into water and was extracted with ethyl acetate. After chromatography of organic matter was obtained 80 mg 11, melting point 143-145°With (ethanol-water); Rf0,41 (toluene-acetone 7/3); NMR (CDCl3) δ of 4.45 (m, 1, SNON), of 0.93 (s, 3, CH3), of 0.90 (t, 3, CH3), 6,62 + 7,10 (AB, 2, H1, 2), 6,53 (d, 1, H4).

Connection 13

To a solution of 29 ml of 1 M allylbromide magnesium in ether was added 80 ml of anhydrous THF. At -50°added 10 g 7α-propyl,16α-allylation-3-O-benzyl ether (12) in 40 ml THF. After stirring for 1/2 h and the mixture was left to warm to room temperature and added into 300 ml feast upon. water NH4Cl. The product was extracted with ethyl acetate and was purified by chromatography on silica compound is hardly to remove stereoisomers with obtaining 7.2 g of the desired 16α on 17α-Gallileo derivative 13.

Rf0,26 (heptane-ethyl acetate 9/1); 17β-allyl-isomer Rf0,45. NMR (CDCl3) δ to 0.88 (t, 3, CH3), is 0.96 (s, 3, CH3), between 6.08 (m, 1, CH-allyl), 5,80 (m, 1, CH-allyl), 4,95-5,20 (m, 4, 2 CH2-allyl), 5,02 (CH2OBz).

The connection 14

Portion 40 mg dichloride

benzylideneacetophenone (catalyst exchange Grubbs) was added to a solution of 450 mg 13 in 10 ml of methylene chloride. After stirring for 1 h was added 400 mg of catalyst. After 12 h the solvent was removed and was replaced with 20 ml of toluene and the mixture was stirred with 5 g of basic aluminum oxide at 60°for absorption of the catalyst. After filtration through celite and washing with toluene and ethyl acetate was received 400 mg of essentially pure 14; Rf0,34(heptane-ethyl acetate 8/2); Rf13 0,45. NMR (CDCl3) δ 5,02 (s, 2, CH3O), 5,97 (s, 2, CH=CH), of 0.97 (s, 3, CH3).

The connection 15

To a solution of 340 mg 14 in 0.5 ml of methylene chloride was added 8 μl of pyridine, of 0.14 ml of 30% aq. H2O2with the subsequent addition of 2 mg of methyltrioxorhenium. After stirring for 1 h epoxidation was completed, resulting in obtaining mainly desired β-epoxide, as well as to a number of α-isomer. The mixture was poured into water and feast upon. Na2S2About3and were extracted with ethyl acetate. After chromatography the raffia received 230 15 mg and 80 mg of unwanted α -epoxide. Rf0,36 (toluene-ethyl acetate 95/5; for comparison: Rfconnection 14: 0,39). NMR (CDCl3) δ 3,31, to 3.38 (m, 2, CH(Oh)CH).

The connection 16

A mixture of 4.2 g of 15 and 350 mg LiAlH4in 20 ml of anhydrous THF was heated under reflux for 2 hours Then the mixture was cooled and successively treated with 1 ml feast upon. water Na2SO4, 28 ml of ethyl acetate and 8 g of Na2SO4. After stirring for 1/2 h at room temperature the reaction mixture was filtered through celite and the filtrate was concentrated and chromatographically obtaining 3.4 g 16, melting point 147-148°, Rf0,20 (toluene-ethyl acetate 8/2; Rf23 0,55). NMR (CDCl3) δ of 4.25 (broad s, 1, SNON), of 0.90 (s, 3, CH3), 0,86 (t, 3, CH3), 5,02 (s, 2, OCH2Ar.

Compound 17: (=7α,16β,17α,22S)-7-propyl-16,24-cyclo-19,21-Generala-1,3,5(10)-triene-3,17,22-triol

A solution of 3.3 g 16 in 200 ml of ethanol was first made in the presence of 300 mg of 5% Pd/C. After completion of the reaction the catalyst was filtered through celite and the solution concentrated. The residue is triturated with ether and then with water to obtain 1.7 g 17; melting point 146-147°, Rfof 0.53 (toluene-ethyl acetate 1/1; Rf16 0,60). NMR (CDCl3) δ 4.26 deaths (broad s, 1, SNON), 7,14, 6,62 (AB, 2, H1, H2), is 6.54 (d, H4).

Compound 18: (=7α,16β,17α,22S)-3,22-diacetoxy-7-propyl-16,24-cyclo-19,21-Generala-1,3,5(10)-triene-ol

100 mg with the unity 17 was dissolved in 1 ml of pyridine and treated with 0.1 ml of acetic anhydride. After stirring for 2 h was added water and the mixture was concentrated. The obtained residue was dissolved in ethyl acetate and was extracted with water and saturated aqueous Panso3. The organic solvent was removed under reduced pressure to obtain the target compound (110 mg). NMR (CDCl3) δ to 0.88 (t, 3H, CH3, propyl); a 1.08 (s, 3H, 18-CH3) ; is 2.09 (s, 3H, acetyl); of 2.28 (s, 3H, acetyl); (d, 1H, H4); 6,83 (dd, 1H, H2).

Evaluation of estrogenic activity in vitro

Compounds were evaluated for their activity against estrogen receptors in the test link and test development, applying α or β estrogen receptor human.

Competitive binding to cytoplasmic estrogen receptor man α or β from recombinant cells of the Chinese hamster ovary (Cho) was used to assess the relative affinity of binding (=RBA) (effectiveness) of the tested compounds compared to (17β)-estradiol (E2for estrogen receptors α or βpresent in the cytosol of recombinant cells SNO, stably transfected estrogen receptor-man α (hERα) or receptor β (hERβ).

Estrogenic and antiestrogenic activity of compounds was determined in bioanalysis in vitro using recombinant cells of the Chinese hamster ovary (Cho), stabilin cotransfection estrogen receptor man α (hERαor β (hERβ), the promoter of the rat oxytocin (RO) and the reporter gene luciferase (LUC). Estrogen agonistic the transactivation (effectiveness) of the tested compounds in the form of stimulation of the TRANS-activation of the enzyme luciferase-mediated estrogen receptor hERα or hERβ, compared with estrogen standard estradiol. Antiestrogenic activity (effectiveness) of the tested compounds, i.e. the ability of the TRANS-activation inhibition of the enzyme luciferase-mediated action (17β)-estradiol on estrogen receptor hERα or hERβ, compared with the standard ICI 164,384 (=(7α, 17β)-N-butyl-3,17-dihydroxy-N-methylestra-1,3,5(10)-triene-7-undecanone).

The results (see table 1).

Table 1
The compound of formula 3R7R116-ring (n=1)or

5-ring (n=0)
The analysis of binding Egα/HβTransactivation Itα/HβThe selective agonist Erα/Erβ
AndNN6-ring8,6/0,35,5/0,227
InN6-ring20/5,216/<0,1>160
C(=11)With2H5N5-ring18/1412/<0,1>120
DWith2H5N6-ring17/8,711,5/0,1>115
E(=171))With3H7N6-ring42/13,926/<0,1>260
FNWith3H75-ring16,3/0,411/0,255
GCH3N7-ring 2,3/<0,02>115
NCH3CH36-ring 30/0,3585
IWith4H9N6-ring 2,9/0,02>145
1)17 is a Hen 41621

The connection 17 in antagonistic tests showed the degree of selectivity Erα/Erβ <0,1/67.

The connection corresponding to the prior art, according to the formula 3, but not containing 22-hydroxy-group containing R7α-propyl, R11hydrogen and 6-membered E-ring with the name (7α,16β,17α)-7-propyl-16, 24-cyclo-19, 21-Generala-1,3,5(10)-triene-3,17-diol, in the analysis of binding possessed ratio ERα/ERβ 15/7 and analysis of agonistic development 0,3/<0,1.

Evaluation of preventive actions on bone loss induced by oophorectomy, rats (antiosteoporosis test)

Introduction

Oophorectomy in rats induces loss of bone mass, which is a consequence of deficiency of estrogen. Introduction estrogen compounds prevents this effect. The test used for the evaluation of compounds against antiosteoporosis activity in rats with oophorectomy (OVX). The effect on bone mass can be assessed using peripheral quantitative computed tomography (pQCT) or by quantitative analysis of radiographs. The level of osteocalcin in plasma and the level of deoxypyridinoline, calcium and phosphate in the urine evaluate bone metabolism. The increase in uterine weight and reduce body weight and thymus reflects the effect of the estrogen.

Test animals

Adults unfertilized females of Wistar rats weighing 225-250 g Genre: Hsd/Cpd: Wu, SPF-bred by Harlan, CPB, Zeist, The Netherlands.

The Expo is iment

On the 1st day of the experiment rats were weighed and distributed to the cells depending on body mass, while the rat with the lowest body weight was placed in the first cell and the rat with the highest weight in the last cell. A processing circuit for rats was chosen arbitrarily (randomly).

Sham surgery and ovariectomy was performed under anesthesia with ether. After recovery from anesthesia within 24 h was introduced to the media, control the connection and test the connection once or twice a day for 4 weeks. During this period, the rats weighed weekly. After 4 weeks conducted the autopsy. At necropsy, rats were anestesiologi ether and took the blood from the abdominal aorta. Allocated both hips, spine L1L2L3L4 (optional), uterus, thymus, liver, kidney, adrenal, thyroid and pituitary. Measurement of mineral density of bone tissue and the geometry of the right femur was performed using pQCT in the day of autopsy on fresh tissues. Mineral density of trabecular bone metaphyses part of the femur (FBMDDIS mg/cm3) were determined using pQCT (peripheral quantitative computed tomography; XART A, Stratec, Birkenfeld, Germany).

Interpretation of results

Oophorectomy leads to a statistically significant decrease in bone density of the distal portion and the volume trabeculoplasty tissue of the femur and to a statistically significant higher levels of osteocalcin in plasma and deoxypyridinoline in the urine (P< 0,05, 2 way ANOVA).

The test compound was considered active when the mean values of bone density in the distal part of the femur were significantly elevated compared with the control group with oophorectomy. The influence of compounds on the level of deoxypyridinoline in urine reflect the effect on bone resorption, the impact on the level of osteocalcin in plasma reflect the impact on the renewal of bone tissue, and could help to understand the mechanism of action.

The minimum active dose (MAD) is the dose, which is achieved average proportional difference in mineral density of trabecular bone between 40 and 60%.

Links

Wronski TJ and C.F. Yen: The ovariectomised rat as an animal model for postmenopausal bone loss. Cells and Materials, Supp. 1 (1991): 69-76.

Yamazaki and I. Yamaguchi H.: Characteristics of an ovariectomised osteopenic rat model. J. Bone Min. Res. 4 (1989): 12-22.

Ederveen A.G.H. and H.J. Kloosterboer: Tibolone is a steroid with a tissue specific hormonal profile, completely prevents ovariectomy-induced bone loss in sexually mature rats. J. Bone &Mineral Research, Vol 14, pp. 1963-1970, 1999.

The result is:

Compound 17 (Org 41621): in the test for osteoporosis oral 30 mg/kg

The connection corresponding to the prior art (7α,16β,17α)-7-propyl-16,24-cyclo-19,21-Generala-1,3,5(10)-triene-3,17-diol: 190 mcg/kg orally.

Evaluation of estrogenic activity in vivo

Estrogenic activity in vivo was determined using the test-Allen-Doisy, described in F. Allen, L.A. Doisy, J. Amer. Med.Assoc., 81, 819-821 (1923).

The result is:

Compound 17 (Org 41621): Allen-Doisy subcutaneous 5 mg/kg, oral 30 mg/kg

The connection corresponding to the prior art (7α,16β,17α) -7-propyl-16,24-cyclo-19,21-Generala-1,3,5(10)-triene-3,17-diol: Allen-Doisy subcutaneously 24 mcg/kg, oral 125 mg/kg

1. Steroid compound of the formula (I)

where the dashed links represent optional double bonds;

R6represents H, =CH2or-CH3or-CH2-CH3;

R7represents H, C1-4-alkyl, C2-5alkenyl or2-5-quinil;

R11represents H, C1-4-alkyl, C2-4alkenyl,2-4-quinil or1-4-alkyliden;

E represents a formed together with the carbon atoms 16 and 17 of the steroidal structure five - semiline ring, and the specified ring is a α zespolony in relation to steroid structure, optionally containing one to two vnutrirossijskikh relations;

or its prodrug.

2. Steroid compound according to claim 1, wherein R6represents H, R7represents a C1-3-alkyl, ring E represents a five - or six-membered ring contains no double bonds and containing hydroxycu the PU in position 22 S stereoconfiguration.

3. Steroid compound according to claim 1 or 2, characterized in that the compound has the formula 2

and the name (7α, 16β,17α, 22S)-7-propyl-16,24-cyclo-19,21-Generala-1,3,5(10)-triene-3,17,22-triol.

4. Steroid compound according to any one of claims 1 to 3 for use in therapy.

5. How selective modification of the activity of estrogen receptors by interacting compounds according to any one of claims 1 to 3 with estrogen receptors.

6. The method according to claim 5, characterized in that the method is used with non-therapeutic purposes.

7. The method according to claim 5 or 6, characterized in that the method is selective modification of the activity of estrogen receptors α by reacting compound according to any one of claims 1 to 3 with estrogen receptors α.

8. The method according to claim 5 or 6, characterized in that the method is selective modification of the activity of estrogen receptors β by reacting compound according to any one of claims 1 to 3 with estrogen receptors β.

9. Steroid compound according to any one of claims 1 to 3 to obtain a medicinal product for selective therapy involving estrogen receptors.

10. Steroid compound according to claim 9, characterized in that the selective therapy involving estrogen receptors, is the prevention of sludge the treatment premenopause or postmenopausal state, cause of complaint.

11. Steroid compound according to claim 9, characterized in that the selective therapy associated with estrogen receptors, represents contraception.

12. Pharmaceutical composition having anti-estrogenic activity, containing the steroid compound according to any one of claims 1 to 3 and a pharmaceutically acceptable excipient.



 

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FIELD: chemical technology, natural materials, medicine, pharmacy.

SUBSTANCE: invention relates to the improved method for preparing betulin from betulinic acid that can be used in preparing anti-tumor and anti-HIV medicinal preparations. Method for preparing betulinic acid involves oxidation of betulin with chrome (VI) oxide in acetic acid to betulonic acid and reduction with sodium boron hydride to betulinic acid. Betulonic acid sodium salt is reduced to betulinic acid and reduction reaction is carried out at room temperature at the concentration of sodium boron hydride 1.0-6.0 wt.-%. Invention provides simplifying method for preparing betulinic acid, reducing its cost and enhancing ecological safety of the process of it producing.

EFFECT: improved preparing method.

1 cl, 4 ex

FIELD: organic chemistry, steroids, pharmacy.

SUBSTANCE: invention describes unsaturated 14,15-cyclopropanoandrostanes of the general formula (I):

wherein R1 means hydrogen atom (H), hydroxy-group (OH); R2 means hydroxy-group (OH), hydrogen atom (H); R3 means hydrogen atom (H), (C1-C10)-alkyl at α- or β-position; R4 means halogen atom (F, Cl, Br) or pseudohalogen group (azide, rhodanide), hydroxy-group (OH), perfluoroalkyl; R5 means (C1-C4)-alkyl; if double bond is at 1,2-position then R4 can mean hydrogen atom (H). Also, invention relates to a method for preparing these compounds and pharmaceutical compositions containing these compounds. Compounds of the formula (I) are compounds eliciting gestagenic and/or androgenic effect.

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

11 cl, 1 tbl, 9 ex

The invention relates to steroids of General formula I

where R1- Oh, (H,H), (H,OR), NOR, where R is H1-6alkyl, C1-6acyl; R2- H or C1-6alkyl, R3- H, or R3- C1-6alkyl, C2-6alkenyl,2-6quinil, possibly substituted with halogen, R4- H, C1-6alkyl or C2-6alkenyl; R5- C1-6alkyl, R5- H, R7- H, C1-6alkyl, R8Is H, OH, halogen;

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FIELD: organic synthesis.

SUBSTANCE: invention provides compounds of general formula I:

in which R1 represents H, halogen, OCH3, or OH; R2 represents (a) -X-(CH2)n-CH2-N(R4)R5, where (i) X represents NH or S; n is integer from 1 to 4; R4 and R5, the same or different, represent C1-C4-alkyl, H, -CH2C≡CH, or -CH2CH2OH; or R4 and R5, together, form nitrogen-containing five- or six-membered cycle or heteroaromatic cycle; or where (ii) X represents O; n is integer from 1 to 4; one of R4 and R5 is CH2C≡CH, or -CH2CH2OH and the other H or C1-C4-alkyl; or R4 and R5, together, form imidazole cycle or nitrogen-containing six-membered cycle or heteroaromatic cycle; or R2 represents (b) -Y-(CH2)nCH2-O-R5, where (i) Y represents O; n is integer from 1 to 4; and R6 represents -CH2CH2OH or -CH2CH2Cl; or where (ii) Y represents NH or S; n is integer from 1 to 4; and R6 represents H, -CH2CH2OH, or -CH2CH2Cl; or R2 represents (c) 2,3-dihydroxypropoxy, 2-methylsulfamylethoxy, 2-chloroethoxy, 1-ethyl-2-hydroxyethoxy, or 2,2-diethyl-2-hydroxy-ethoxy; R3 represents H. halogen, OH, or -OCH3. Claimed compounds are novel selective estrogen receptor modulators. Invention also discloses pharmaceutical composition and a method for production of tissue-specific estrogenic and/or antiestrogenic effect in patient, for whom indicated effect is required.

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19 cl, 7 tbl, 11 ex

The invention relates to medicine

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FIELD: medicine, cosmetology.

SUBSTANCE: one should detect the scope of eruption, the availability of psychoemotional disorders, then, by taking into consideration patient's sex it is necessary to calculate Monakhov's therapeutic acne index (MTAI) by a certain formula to prescribe therapy by taking into account the index obtained. The present innovation provides individual approach to prescribing anti-acne therapy that, in its turn, enables to shorten the terms of disease, decrease the number of jatrogenic complications and prolong remission period.

EFFECT: higher efficiency of anti-acne therapy.

13 ex, 4 tbl

FIELD: organic chemistry, medicine, pharmacy.

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(5) ,

(6) ,

(17) , (18) , (19) , (20) , (23) , (25) and (26) wherein R3 in formula (6) represents a direct or branched group (C1-C5)-alkyl group; R8 in formulae (18) and (20) represents a direct or branched (C1-C5)-alkyl group, a direct or branched (C2-C5)-alkenyl group or a direct or branched (C2-C5)-alkynyl group; in formula (23) each R21, R22, R23 and R24 represents independently hydrogen atom, a direct or branched (C1-C5)-alkyl group, a direct or branched (C1-C7)-halogenalkyl group, halogen atom or acyl group; in formulae (25) and (26) X represents halogen atom; or enantiomers of compound, or hydrates, or pharmaceutically acceptable salts of compound, or its enantiomers. Also, invention relates to a pharmaceutical composition containing indicated compound as an active component and to a therapeutic agent used against breast cancer based on thereof.

EFFECT: valuable medicinal properties of compounds.

10 cl, 2 tbl, 39 ex

FIELD: medicine.

SUBSTANCE: before applying substitute hormonal therapy (SHT) on should evaluate antithrombogenic activity of vascular wall in women. For this purpose one should determine quantitative values of ADP-induced aggregation of thrombocytes, activity of antithrombin III in blood and fibrinolytic blood activity both before and after "cuff"-test. Then one should detect the indices calculated as the ratio of mentioned values both before and after carrying out the mentioned test. If mentioned indices are decreased against the norm by 20-40% women should be prescribed to undergo SHT at additional introduction of aspirin and supradin. The method provides prophylaxis of cardio-vascular diseases in this category of female patients due to correcting affected functional activity of vascular endothelium.

EFFECT: higher efficiency of prophylaxis.

1 cl, 1 ex, 4 tbl

FIELD: organic chemistry, steroids, pharmacy.

SUBSTANCE: invention relates to a new type of selective estrogens comprising steroid structure of the general formula (I) with nonaromatic ring A and free of bound hydroxyl group at carbon atom 3 wherein R1 means hydrogen atom (H), (C1-C3)-alkyl or (C2-C3)-acyl; R2 means hydrogen atom (H), α-(C1-C4)-alkyl, α-(C2-C4)-alkenyl or α-(C2-C4)-alkynyl; R3 means hydrogen atom (H) or (C1-C4)-alkyl at position 16 of steroid structure; R4 means ethynyl; R5 means hydrogen atom (H), (C1-C3)-alkyl or (C2-C3)-acyl; R6 means (C1-C5)-alkyl, (C2-C5)-alkenyl, (C2-C5)-alkynyl being each of that is substituted optionally with chlorine or fluorine atom; dotted line means the optional double bond. Compounds of the formula (I) elicit the selective affinity to ERα-receptors.

EFFECT: valuable properties of compounds and composition.

4 cl, 3 sch, 1 tbl, 8 ex

FIELD: medicine, gynecology.

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EFFECT: higher efficiency of therapy.

31 cl, 1 dwg, 1 ex, 2 tbl

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