Steroid compounds (options), pharmaceutical composition, method for the treatment of nervous disorders (options) and the method of induction of anesthesia the animal

 

(57) Abstract:

Describes the steroid compounds of General formula I, where (a) R4- 3'-methyl-but-3'-EN-1'-inyl or cyclopropylethyl, R5is methyl, R6- acetyl, dashed line means a single bond; b) R4- trifluoromethyl, R5is methyl, R6- acetyl, dashed line means a single or double bond and when a single link, R6- hydroxyacetic, a R5is hydrogen, R4may mean vermeil; R4- ethinyl, R5is methyl, R6- hydroxymethyl or hydroxyacetic, dashed line means a single bond, or a physiologically acceptable complex 21-esters, 3,20-or diesters 3,21-diesters, or where a)6- hydroxyacetic, R5is hydrogen when R4- trifluoromethyl and the connection represented in the form of 21-hemisuccinate sodium; b) R6- hydroxyacetic, R5is methyl, and R4accepts the following values: hydrogen, in the case where the compound represented by bohemiacallada; ethinyl, in the case of a 21-hemisuccinate; trifluoromethyl in the case of a 21-hemifumarate sodium, methyl 21-succinate, 21-propionate; and R4is methyl, R5is methyl, R6- isopropylidenediphenol, and the dashed line oznaczanie for the treatment of stress, anxiety, epileptic seizures, mood disorders, premenstrual syndrome (PMS) and postpartum depression (PND) and for induction of anesthesia. 9 C. and 6 C.p. f-crystals, 8 PL.

THE TECHNICAL FIELD TO WHICH THE INVENTION RELATES

The invention is devoted to methods, compositions and compounds for modulation of excitability of the brain of animals with receptor-aminobutyric acid A (GABAA) - chloride ionogennogo complex (GRC). In particular, the present invention is directed to methods, compositions and compounds for modulation of excitability of the brain by binding site non-steroidal receptor on GRC.

ART

The excitability of the brain is defined as the level of arousal of the animal, condition ranging from coma to convulsions, and are subject to various neurotransmitters. In the General case, the neurotransmitter responsible for regulating the conduction of ions across the membranes of neurons. At rest, the membrane of a neuron has the potential (or membrane voltage) approximately -80 mV, the internal portion of the cell is negatively charged compared to the outside of the cell. The potential (voltage) is the result of runaway are in the presynaptic vesicles and are released under the influence of the action potential of a neuron. When entering the synaptic deepening chemical mediator of irritation, such as acetylcholine, will cause membrane depolarization (change of potential from -80 mV to -50 mV). This effect is transmitted to postsynaptic nicotinic receptors, which are stimulated by acetylcholine to increase the permeability of the membrane in relation to ions of Na+. Reduced membrane potential stimulates neural excitation in the form of a postsynaptic potential actions.

In the case of GRC effect of excitability of the brain is carried by the neurotransmitter GABA. GABA has a strong impact on the overall excitability of the brain, because up to 40% of brain neurons use GABA as a neurotransmitter. GABA regulates the excitability of individual neurons by regulating the conductance of chloride ions through the membrane of neurons. GABA interacts with your website recognition at GRC to increase flow of chloride ions in the direction of the electrochemical gradient GRC in a cage. The increase in the content of this anion inside the cell causing hyperpolarization of transmembrane potential, making the neuron less sensitive to the stimulating signals (for example, reduced neuronal excitability). Druggee).

It is well known that GRC is responsible for anxiety, pripadochnaya activity and sedative effect. Thus, GABA and medicines, which act as GABA or facilitate the action of GABA (e.g., therapeutically used barbiturates and benzodiazepines (BZs), such as valium) exert their therapeutic effect by interaction with specific regulatory sites on GRC.

It is also shown that a series of steroid metabolites interact with GRC to change the excitability of the brain (Majewska, M. D. et al., Science (1986), 232:1004-1007; Harrison, N. L. et al., J. Pharmacol. Exp. Ther., 1987, 241: 346-353). Until the present invention the possibility of therapeutic use of these steroid metabolites was not known to the researchers working in this area, due to incomplete understanding of the activity and scope. The present invention is partially related to the use of pharmaceutical knowledge gained from a better understanding of the activity and scope of certain steroid compounds.

It was demonstrated that the ovarian hormone progesterone and its metabolites have a strong effect on the excitability of the brain (Backstrom T. et al., Acta Obstet. Gynecol. Stand the possession of progesterone and its metabolites vary depending on the phase of the menstrual cycle. It was documented that progesterone and its metabolites are reduced before the onset of menstruation. Monthly repetition of certain physical symptoms before menstruation has also been documented. These symptoms, which were associated with premenstrual syndrome (PMS) include stress, anxiety, and migraine headaches (Dalton K., Premenstrual Syndrome and Progesterone Therapy, 2nd edition, Chicago, Chicago yearbook, 1984). Patients with PMS have a monthly recurrence of symptoms, which prisutstvuyuthie menstruation and is missing after menstruation.

Similarly the decrease in progesterone also temporarily correlated with increased frequency of seizures in women epileptics (e.g., menstrual epilepsy; J. Laidlaw, "Catamenial epilepsy", Lancet, 1956, 1235-1237). A more direct correlation was observed in the reduction of metabolites of progesterone (Rosciszewska et al., J. Neurol. Neurosurg. Psych., 1986, 49:47-51). In addition, in patients with primary generalised minor epilepsy time seizures correlated with the time of the symptoms of premenstrual syndrome (Backstorm T. et al., J. Psychosom. Obstet. Gynecol., 1983, 2:8-20). It was found that steroid hypertension is effective in the treatment of patients with epileptic seizures, correli the m with low levels of progesterone, is postnatal depression (PND). Immediately after birth, the levels of progesterone drop sharply, leading to the beginning of the PND. The PND symptoms range from mild depression to psychosis requiring hospitalization; PND is associated with a state of great anxiety and irritability. PND-associated depression cannot be treated by classical antidepressants, and women with the syndrome PND increases the probability syndrome PMS (K. Dalton, 1984).

Together, these observations suggest a significant role of progesterone and hypertension and, more specifically, their metabolites in the homeostatic regulation of excitability of the brain, which manifests as increased epileptic activity or symptoms associated with the menstrual epilepsy, PMS and PND. The correlation between reduced levels of progesterone and symptoms associated with PMS, PND and menstrual epilepsy (Backstorm et al., 1983; Dalton K., 1984), determined the use of progesterone in their treatment (Mattson et al., in Advances in epileptology: XVth Epilepsy International Symposium, Raven Press, New York, 1984, 279-282, 1984 and K. Dalton). However, progesterone is not always constantly effective in the treatment of the above-mentioned syndromes. For example, no link is 1986, 290:16-17).

Publications and references referred to above and hereinafter in this description, incorporated herein as references.

THE INVENTION

The present invention relates to a method, compositions, and compounds for modulating the excitability of the brain. In particular, the invention concerns the use of 3-hydroxylated steroid derivatives acting on the newly identified site of the GR complex, to modulate the excitability of the brain in such a way as to reduce stress, anxiety, insomnia, worsening of mood (e.g. depression), which are GR-active agents, as well as epileptic activity. Compositions and compounds effective for such treatment are covered by the present invention.

Used in the invention and forming part of the invention compounds are modulators of excitability of the Central nervous system, which follows from their ability to regulate the channels of chloride ions associated with the GABA receptor. Experimentally applicants found that the compounds according to the invention and used in the invention possess anticonvulsive activity against alarm condition, such action Investec complex GR.

Relationship of endogenous metabolites of progesterone with the processes associated with reproduction (the estrus cycle and pregnancy), is well established (Marker, R. E., O. Kamm, and R. V. McGrew, "Isilation of epi-pregnanol-3-one-20 from human pregnancy urine", J. Am. Chem. Soc., 1937, 59:616-618). However, until the present invention was not known to treatment by modulation of excitability of the brain through the use of metabolites of progesterone. Therefore, the present invention is directed to methods, compositions and compounds for the treatment of diseases by modulation of excitability of the brain with the use of the compounds according to the invention. Examples of diseases that are treated according to the present invention, are epilepsy, anxiety, premenstrual syndrome (PMS), post-Natal depression (PND), mood disorders (such as depression), which are GR-active agents, as well as insomnia. Compounds according to the invention can also be used for anesthesia.

INFORMATION CONFIRMING THE POSSIBILITY OF CARRYING OUT THE INVENTION

The compounds according to the invention and used in it are derived the various 3-hydroxylated-pregnan-20-ones, 3-21-pregnandiol-20-ones, 3-20-pregnancylow and 3-hydroxylated androstan osvitnye, which is considered as prodrugs. The expression "prodrug" means a derivative known drugs direct action that is derived which has improved characteristics receipts and therapeutic action compared to the drug and become active drug enzymatic or chemical means; see Notari, R. E., "Theory and Practice of Prodrag Kinetics", Methods in Enzymology, 1985, 112: 309-323; Bodor, N., "Novel Approaches in Prodrug Design", Drugs of the Future, 6(3):165-182 (1981) and H. Bundgaard, "Design of Prodrugs: Bioreversible Derivatives for Various Functional Groups and Chemical Entities," in Design of prodrugs (H. Bundgaard, ed.), Elsevier, New York (1985). It should be noted that some of the synthesized derivatives constituting part of the present invention may not be true prodrugs, because in addition to the above characteristics, they also have their own activity. However, under this proposal they will be referred to as prodrugs.

Our research (Gee, K. W. et al., European Journal of Pharmacology, 1987, 136: 419-423) showed that 3-gidroksilirovanii steroids used in the invention, by several orders of magnitude more active than other previously described (M. D. Majewska et al., 1986 and Harrison, N. L. et al., 1987) as modulators of complex GR. Majewska, M. D. et al. and Harrison, N. L. et al. pokazyvauschie. Our experimental data in vitro and in vivo show that the high activity of these steroids causes their therapeutic use for the modulation of excitability of the brain through a complex GR. The most active steroids used in this invention include derivatives of the major metabolites of progesterone and hypertension. These steroids can be in the specific case used to modulate the excitability of the brain during stress, anxiety, insomnia, mood disorders (e.g. depression) that can be manipulated GR-active agents, as well as in epileptic disorders therapeutically favorable way. Moreover, we showed that these steroids interact with a unique site on the GR complex, which differs from other known sites of interaction (such as barbiturates, BZ and GABA), which was earlier achieved therapeutically beneficial effects on stress, anxiety, sleep, mood disorders and epileptic disorders (Gee, K. W. and Yamamura, H. I. In Central Nervous System Disorders, 1985, 123-147, D. C. Horvell ed.; Lloyd K. G. and P. L. Morselli in Psychopharmacology: The Third Generation of Progress, 1987, 183-195, H. Y. Meltzer, ed., Raven Prss, N. Y.). These compounds are desirable because of about what edenia).

Steroid derivatives according to the invention have one of the following structural formula:

Formula 1

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Formula 2

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Formula 3

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Formula 4

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Formula 5

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Formula 6

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The present invention also includes pharmaceutically acceptable esters and salts of compounds of Formulas 1-6, including acid salt. It is believed that the 3-hydroxyl can also be hidden in the form of a pharmaceutically acceptable ether complex due to the fact that the ether group will be split when converting into a dosage form. Such compounds are considered biodegradable esters.

DEFINITION

In accordance with this invention the terms used have the following meaning, unless a different meaning.

The term "alkyl" means a saturated aliphatic groups, including non-branched chain, branched chain, cyclic groups, all of which may be substituted. Suitable alkyl groups include methyl, ethyl and similar groups and can be substituted.

The term "alkenyl" means unsaturated groups, which contain at least one carbon-carbon double bond, and includes an unbranched chain, nasyshennye hydrocarbon group, which contain at least one carbon-carbon triple bond, and includes an unbranched chain, branched chain and cyclic groups, which may have additional unsaturation. In position 3 - alkyline group may be substituted halogenated or dehalogenating C1-radical, saturated or unsaturated, halogenated or dehalogenating non-branched C2-C6radicals, cyclic (cycloalkyl) C3-C6radicals or aromatic C5-C6-radical or a 4-, 5 - or 6-membered heterocyclic radical that is attached to atom C - or N - containing 1, 2 or 3 heteroatoms selected from the group consisting of oxygen, nitrogen or sulfur, excluding heterocyclic radicals with two or more adjacent atoms O or S. Preferred alkenylamine groups are groups containing from two to four carbon atoms.

The term "alkoxy" refers to a simple ether group-OR, where R is alkyl.

The term "aryloxy" refers to a simple ether group-OR, where R is aryl.

The term "aryl" means an aromatic group that contain at least one cycle with conjugated PI-electron sistency aryl" means a group in which cyclic atoms of the aromatic cycle are carbon atoms. Carbocyclic aryl groups include phenyl and raftiline group which may be substituted. Substituted phenyl preferably contains from one to three substituents, mainly such as lower alkyl, amino, hydroxyl, lower alkoxyl, halogen, lower acyl and the nitrogroup.

The term "aralkyl" means alkyl group, substituted aryl group. Suitable kalkilya groups include benzyl and the like groups and can be substituted.

The term "dialkylamino" refers to NRR 'where R and R", independently of one another represent lower alkyl groups, or together they form the remainder morpholino group. Suitable group dialkylamino include dimethylamino, diethylamino and morpholino group.

The term "acyl" refers to alkanoyloxy the group-C(O)R, where R is alkyl, alkenyl, quinil, aryl or aralkyl.

The term "possibly substituted" or "substituted" refers to groups substituted by one to three substituents, independently from each other selected from lower alkyl, aryl, alkenyl, quinil, alkoxyl, amino, tigroup, halogen, haloalkyl, trihalogen-alkyl, acyl, the current from one to four carbon atoms. Such groups can be unbranched, branched or cyclic.

The term "pharmaceutically acceptable esters or salts" refers to esters or salts of Formulas 1 to 6, obtained from the combination of the compounds according to the invention and an organic or inorganic acid.

Examples of substituents that can be used in the compounds of Formula 1, are the following:

R is hydrogen, lower alkyl, lower alkenyl or lower quinil;

R1is methylene, hydroxymethyl or cyano; and their physiologically acceptable 3-esters, 20-esters and 3,20-diesters; provided that if R1- is cyano, R is not hydrogen.

However, in the method according to the invention R1may be a cyano, where R is hydrogen.

A preferred group of compounds of Formula 1 are compounds in which R is hydrogen.

Another group of preferred compounds are compounds of Formula 1, where R1means cyano.

Examples of preferred compounds of Formula 1 include, but are not limited to, the following: 3-hydroxy - 17-methylene-5-androstane; 3-hydroxy-17 - hydroxymethyl-5-androstane; or 3-hydroxy -17(20)(Z)-ethoxymethylene - 5-Andes is the following:

R is hydrogen, halogen or lower alkoxy;

R1alkenyl, quinil, alkoxyalkyl, haloalkoxy, trifluoromethyl, azidoethyl, cianelli or monohalogen-methyl;

R2- hydrogen, ketogroup or 11-dialkylamino;

R3- -acetyl group, ketal-hydroxyazetidine group, TRIFLUOROACETYL, -hydroxyacetic-hydroxyacetic-17-hydroxyl, -methoxyethylamine, (ethoxy)-methyl-2'-metilurazil, -(1'-hydroxyethyl), -(1'-hydroxypropyl), -(2'-hydroxyisopropyl)- succinicacid-hydroxyacetic succinate sodium-acetoxyacetyl-sulfoxylate-methylacyl-haloacetic, i.e.- chloroacetyl-ethinyl or-ethyl, together with the 17th carbon atom to which the specified ethyl group is attached, and an oxygen atom form 17(20)apachegroup; and their physiologically acceptable complex 3-esters, 20-ethers, 21-esters, 3,20-diesters and 3.21-diesters; provided that, if R2means 11-N,N-dialkylamino, R is not hydrogen.

A preferred group of compounds of Formula 2 are compounds in which R is hydrogen or lower alkyl. More preferred are compounds where R is hydrogen.

Another group of preferred compounds of imperialism and monoelemental. More preferred are compounds where R1- C2-C4alkenyl, C2-C4-quinil or trifluoromethyl. Most preferred are compounds where R1means ethinyl or trifluoromethyl.

An additional group of preferred compounds are the compounds of formula 2 where R3means acetyl, hydroxyalkyl, hydroxyacetic or their esters with physiologically acceptable acids. More preferred are compounds where R3- acetyl or beta-succinicacid. Most preferred are compounds where R3is acetyl.

Examples of preferred compounds include, but are not limited to, the following: 3-hydroxy-17-ethinyl-5-androstane; 3-ethynyl-3-hydroxy-5-pregnan-20-he; 3-hydroxy-3 -(2'-propenyl)-5-pregnan-20-he; 3-ethynyl-3-hydroxy-5-pregnan-20-he; 3-chlorethyl-3-hydroxy-5-pregnan-20-he;

3-ethinyl-3-hydroxy-5-pregnan-20-he, 3-hydroxy-3 - ethinyl-5-pregnan-20-he, 3,20-dihydroxy-3-ethinyl-5-pregnan, 3,21-dihydroxy-3-ethinyl-5-pregnan-20-he, 3-(3'-bromo-1-PROPYNYL)-3-hydroxy-5-pregnan-20-he, 3, 21-dihydroxy-3-ethinyl-5-pregnan-20-he, 21-acetate, 3-ethinyl-3-hydroxy-5-pregnan-11,20-dione, 3, 21-dihydroxy-3-ethinyl-5-pregnan-20-21 he-gemisch the hydroxy-hydroxy-3-(2' -PROPYNYL)-5-pregnan-20-he, 3,20-dihydroxy-3-ethinyl-5-pregnan, 3,20-dihydroxy-3-ethinyl-5-pregnan, 3-ethynyl-3-hydroxy-21-methoxy-5-pregnan-20-he; 3-hydroxy-3-chloromethyl-5-pregnan-20-he, 3-hydroxy-3-trifluoromethyl-5-pregnan-20-he, 3-methyl bromide is 3-hydroxy-5-pregnan-20-he, 3-hydroxy-3-iodomethyl-5-pregnan-20-he, 3-hydroxy-3-trifluoromethyl-5-pregnan-20-he, 3-hydroxy-3-trifluoromethyl-5-pregnan-20-he, 3,21-dihydroxy-3-trifluoromethyl-5-pregnan-20 he 21-acetate, 3,21-dihydroxy-3-trifluoromethyl-5-pregnan-20-he, 3-hydroxy-3-trifluoromethyl-5-pregnan-11,20-dione, 3, 21-dihydroxy-3-trifluoromethyl-5-pregnan-20-he, 21-hemisuccinate, sodium salt, 3-hydroxy-3-trifluoromethyl-5-pregnen-17(20)-ene, 3,20-dihydroxy-21-ethyl-5-pregnan, 3,20-dihydroxy-21-methyl-5-pregnan, 3,20-dihydroxy-2-isopropoxy-5-pregnan, 3,20-dihydroxy-2-ethoxy-5-pregnan, 3,20-dihydroxy-2-n-propoxy-5-pregnan,

3,20-dihydroxy-3-methyl-5-pregnan, 3,20-dihydroxy-3-ethinyl-5-pregnan, 3,20-dihydroxy-3-methyl-5-pregnan, 3,20-dihydroxy-3,20-dimethyl-5-pregnan, 3,20-dihydroxy-3 21-dimethyl-5-pregnan, 3,20-dihydroxy-3-ethinyl-5-pregnan, 3,20-dihydroxy-3-methyl-5-pregnan, 3,20-dihydroxy-3-ethinyl-5-pregnan, 3-hydroxy-3-methoxymethyl-5-pregnan-20-he, 3-(ethoxymethyl)-3-hydroxy-5-pregnan-20-he, 3-(1'-hexenyl)-3-hydroxy-5-pregnan-20-he, 3-azidomethyl-3-hydroxy-5-pregnan-20-the -3-hydroxy-5-pregnan-2-it, 3-(methylbut-2-EN-1-inyl)-3-hydroxy-5-pregnan-20-he, 3-(1'-heptenyl)-3-hydroxy-5-pregnan-20-he, 3-cyclopropylmethyl-3-hydroxy-5-pregnan-20-he, 3-(1'-octenyl)-3-hydroxy-5-pregnan-20-he, 3-cyclopropylmethyl-3-hydroxy-5-pregnan-20-he and 3-cyclopropylmethyl-3-hydroxy-5-pregnan-20-he.

Examples of substituents that can be used in the compounds of Formula 3 are as follows:

R is hydrogen, lower alkyl, lower alkenyl or lower quinil;

R1- -acetyl, ketal-gidroksietilimino group, TRIFLUOROACETYL, -hydroxyacetic-hydroxyacetic-17-hydroxyl, -methoxyethylamine, (ethoxy)-methyl-2'-metilurazil, -(1'-hydroxyethyl), -(1'-hydroxypropyl), -(2'-hydroxyisopropyl)- succinicacid-hydroxyacetic succinate sodium,

-acetoxyacetyl-sulfoxylate-methylacyl, -chloroacetyl-ethinyl or-ethyl, which together with the 17th carbon atom to which the specified ethyl group is attached, and an oxygen atom form 17(20)apachegroup; and their physiologically acceptable complex 3-esters, 20-ethers, 21-esters, 3,20-diesters and 3.21-diesters; provided that said compound is not 3-hydroxy-5-preg-9(11)-ene-20-he, 3,21-dihydroxy-5-preg-9(11)-ene-20-one or 21-acetoxy-3-hydroxy-5-preg-9(11)-ene-20 - he or E. the R means hydrogen or quinil. More preferred are compounds in which R is hydrogen.

An additional group of preferred compounds are the compounds of Formula 3, where R1means acetyl, hydroxyalkyl, hydroxyacetic or their esters with physiologically acceptable acids. More preferred are compounds where R1means acetyl or beta-succinicacid. Most preferred are compounds where R1is acetyl.

Examples of preferred compounds include, but are not limited to, the following: 3-hydroxy-3-trifluoromethyl-5-pregn-9(11)-ene-20-he; and 3-hydroxy-3-methyl-5-pregn-9-EN-20-he.

Examples of substituents that can be used in the compounds of Formula 4 are the following:

R is hydrogen, lower alkyl, lower alkenyl or lower quinil;

the value of R1- see Formula 3: and their physiologically acceptable complex 3-esters, 20-ethers, 21-esters, 3,20-diesters and 3.21-diesters; provided that the specified connection cannot be 3-hydroxy-5-preg-11-EN-20-one or its 3-ether derivative.

A preferred group of compounds of Formula 4 are compounds in which R denotes hydrogen, trifluoromethyl or lower quinil. More is respectful of compounds are the compounds of Formula 4, in which R1means acetyl, hydroxyalkyl, hydroxyacetic or their esters with physiologically acceptable acids. More preferred are compounds in which R1means acetyl or beta-succinicacid. Most preferred are compounds where R1is acetyl.

Examples of preferred compounds include, but are not limited to, the following: 3-hydroxy-3-trifluoromethyl-5-pregn-11-ene-20-he; 3,20-dihydroxy-5-pregn-11-ene; 3-hydroxy-3-methyl-5-pregn - 11-ene-20-he; and 3,20-dihydroxy-5-pregn-11-ene, 3-ethinyl-3-hydroxy-5-pregn-11-ene-20-he.

Examples of substituents which may be the compounds of Formula 5 are as follows:

R is hydrogen, lower alkyl, lower alkenyl or lower quinil;

R1- beta-formyl, methylene beta-hydroxymethyl, methoxymethyl or beta-cyano and their physiologically acceptable complex 3-esters, 20-esters and 3,20-diesters.

Examples of preferred compounds include, but are not limited to, the following: 3-hydroxy-17-formyl-5-19-norandrosterone and 3-hydroxy-17(20)(Z) -ethoxymethylene-5-19-norandrosterone.

Examples of substituents that can be used in the compounds of Formula 6, are the following:

R-crystals 3 and their physiologically acceptable complex 3-esters, 20-ethers, 21-esters, 3,20-diesters and 3.21-diesters.

A preferred group of compounds of Formula 6 are compounds in which R signifies lower alkenyl and lower quinil or trifluoromethyl. More preferred are compounds, in which R means a lower quinil or trifluoromethyl.

An additional group of preferred compounds are the compounds of Formula 6, in which R1means acetyl, hydroxyalkyl, hydroxyacetic or their esters with physiologically acceptable acids. More preferred are compounds in which R1means acetyl or beta-succinicacid. Most preferred are compounds in which R1means acetyl.

Further preferred compounds are compounds of Formulas 1 to 6, which are esters by hydroxyl group at positions 3 and/or 20. The preferred esters are the esters of these compounds and the corresponding acids: acetic, propionic, maleic, fumaric, ascorbic, pipelinewall, succinic, glutaric, bitmeyen-salicylic, methanesulfonate, ethane-di-sulfonic acids, oxalic, tartaric, salicylic, citric, gluconic, takenaway, glico is Oh, glycine and other amino acids, phosphoric, sulfuric, glucuronic and 1-methyl-1,4-dihydrocodeinone.

Another more preferred group of compounds includes those compounds of Formulas 1 to 6, in which 3-Deputy is ethinyl or trifluoromethyl, and attached to the 17-th position of the steroid is acetyl, -(1'-hydroxyethyl) or 17(20)(Z)-ene.

Examples of preferred compounds include, but are not limited to, the following: 3-hydroxy-3-methyl-5-19-norpregna-17(20)(Z)-ene, 3-hydroxy-19-nor-5-pregnan-CIS-17(20)(Z)-ene, 3,20-19-norpregna-17(20)(Z)-ene, 3-hydroxy-3-methyl-5-19-norpregna-17(20)(Z)-ene, 3-ethinyl-3-hydroxy-5-19-norpregna-17(20)(Z)-ene and 3-hydroxy-3-trifluoromethyl-5-19-norpregna-17(20)(Z)-ene.

Also preferred are the following compounds: 3-hydroxy-3-trifluoromethyl-19-nor-5-pregnan-20-he, 3-hydroxy-3-trifluoromethyl-19-nor-5-pregnan-20-he, 3-hydroxy-3-trifluoromethyl-5-19-norpregna-17(20)(Z)-ene, 3, 21-dihydroxy-3-trifluoromethyl-19-nor-5-pregnan-20-he 3,21-dihydroxy-3-trifluoromethyl-19-nor-5-pregnan-20-he

21-hemisuccinate and its sodium salt.

Also preferred are the following compounds: 3,20-dihydroxy-19-nor-5-pregnan; 3-hydroxy-3-trifluoromethyl-19-nor-5-pregnan-20-he; 3-hydroxy-3-methyl-5-19-norpregna-20-he; 3-Atini is,21-dihydroxy-3-trifluoromethyl-19-nor-5-pregnan-20-21 he-hemisuccinate and its sodium salt.

The most preferred compounds in this invention are the following:

3-hydroxy-3 -(3'-methyl-but-3'-EN-1'-inyl)-5-pregnan-20-he;

3-hydroxy-3 -(3'-methyl-but-3'-EN-1'-inyl)-5-pregnan-20-he;

3-hydroxy-3-trifluoromethyl-5-pregnan-20-he;

3-hydroxy-3-trifluoromethyl-5-pregn-11-ene-20-he;

3,21-dihydroxy-3-trifluoromethyl-19-nor-5-pregnan-20-he;

3-(cyclopropyl)ethinyl-3-hydroxy-5-pregnan-20-he;

3,20-dihydroxy-3-ethinyl-5-pregnan;

3,21-dihydroxy-3-ethynyl-5-pregnan-20-it; and

3, 21-dihydroxy-3-trifluoromethyl-5-pregnan-20-he; or a physiologically acceptable complex 3-esters, 20-ethers, 21-esters, 3,20-or diesters 3,21-diesters of these compounds.

Also most preferred are esters according to the invention:

3, 21-dihydroxy-3-trifluoromethyl-19-nor-5-pregnan-20-21 he-hemisuccinate, sodium salt;

3,20-dihydroxy-21-methyl-5-pregnan, bis-hemisuccinate;

3,21-dihydroxy-3-ethynyl-5-pregnan-20-he, 21-hemisuccinate;

3,21-dihydroxy-3-methyl-5-pregnen-20 he 21-acetate;

3,21-dihydroxy-3-trifluoromethyl-5-pregnan-20-21 he-hemifumarate, sodium salt;

3, 21-dihydroxy-3-trifluoromethyl-5-pregnan-20-he, methyl-21-succinate;

3,21-dihydroxy-3-trifluoromethyl-5-pregnan-20-AOI-3-ethinyl-5-pregnan-20-21-hemisuccinate and

N-(3,dihydroxy-3-methyl-5-pregnen-20-alidin)ethanolamine.

The following synthetic methods and examples aimed at obtaining the compounds forming part of the invention and used in the present invention.

METHODS FOR OBTAINING COMPOUNDS

Compounds according to the invention can be obtained by any known method, namely using known techniques, such as described in the "Steroid Reactions" Djerassi, published in 1963 Holden-Day, Inc., San Francisco, or "Organic Reactions in Steroid Chemistry" Fried and Edwards, published in 1972, Van Nostrand-Reinhold Co., New York.

Compounds according to the invention can be obtained by any suitable known or developed in the present method.

THE MAIN WAYS

20-hydroxypregn were obtained by reduction of 20-getpregnant conventional reducing agents.

21-Hemisuccinate were obtained from derivatives of pregnan-20-ones, which were first problemyour molecular bromine, resulting in a received corresponding 21-pompignan. Bromo derivatives were then subjected to interaction with various dibasic acids such as succinic chilota, in the presence of the amine, resulting in a received 21-hydroxy-when you.

Esters can be obtained by known reactions interaction of the hydroxyl group of the above compounds with an organic acid, acid halide, acid anhydride or a complex ether, the organic acid is, for example, acetic, propionic, maleic, fumaric, ascorbic, Emelyanova, succinic, glutaric, biotranslation, methanesulfonate, ethane di-acid, oxalic, tartaric, salicylic, citric, gluconic, Takanawa, glycolic acid, para-aminobenzoic, spartanovka, glutamine, gamma-aminobutyric, alpha-(2-hydroxyethylamino)-propionic, glycine and other alpha-amino acids, phosphoric, sulfuric, glucuronic and 1-methyl-1,4-digitaleconomy acid.

3-VICE

Halogen-methyl

3-monohalomethanes compounds according to the invention can be obtained by treating 3-Spiro-2'-oxiranes steroid source of ions of a halogen in an inert solvent, for example, a halide of Tetramethylammonium in toluene and preferably a source of protons, for example acetic acid.

Saturated or unsaturated alkyl

Other 3-substituted steroids can be obtained by adding the ORGANOMETALLIC reagent is ü protected. Thus, 3-alkyline derivatives can be obtained using acetylide lithium in an inert solvent or reagent obtained "in situ" from 1,2-dibromoethylene and butyl lithium, as the ORGANOMETALLIC reagent. Similarly, the compounds of Formula 1, where R is alkenyl, can be obtained by the reaction of 3-ketosteroid with vinyl ORGANOMETALLIC reagent, such as vinylmania. Compounds in which the unsaturation is eliminated in the reaction, such as allylanisole, can also be used to obtain compounds containing 3-alkenylphenol group. Similarly, the use of alkyl Grignard reagent, such as methylmagnesium will result in 3-alkyl derivatives.

Trifluoromethyl

Triptoreline group can be obtained by the reaction of 3-ketosteroid with trimethylchlorosilane catalyzed by fluoride ion.

21-OXIDIZED DERIVATIVES

Oxidation of 21-methyl leads to compounds, which lead

Various derivatives of this type can be obtained as a result of chain reactions, in which pregnan-20-it oxidizes leads to compounds, which lead to obtain 21-acetoxypropionyl, with subsequent hydrolysis of the acetate to obtain 21-alcohol and a suitable acylation is ü the hydrogen of the hydroxyl group, such as methanesulfonate.

Banished-17-ENES

These compounds can be obtained by the reaction of 17-ketosteroid with the Wittig reagent, such as ylides obtained by treatment of methyl n-propyltrimethylammonium bromide strong base such as tert-butyl potassium.

3,20-diols

Pregnan-3,20-diol can be obtained by the reaction of joining borohydride, such as DIBORANE, the double bond of pregnen-17-ene with subsequent oxidation of the obtained boron compounds, for example, hydrogen peroxide in an alkaline medium to obtain a 20-Ola. Alternatively, pregnan-3,20-diols can be obtained by reduction of the group of 20-he in a group of 20-ol. Suitable reactants are the hydrides, such as sodium borohydride or solutions of metals, such as sodium n-propanol and similar.

EXAMPLES

EXAMPLE 1

a. 3-HYDROXY-5-PREGNAN-20-HE, 20-KETAL

A mixture of 3-hydroxy-5-pregnan-20-she (10.8 g, 34 mmol), ethylene glycol (45 ml) and triethylorthoformate (30 ml) was stirred at room temperature for 5 minutes Then add p-toluensulfonate acid (200 mg) and continue stirring at room temperature for 1.5 hours Received a thick paste, pour in the solution nasishenorozoviy product is dissolved in CH2Cl2(350 ml) and dried over anhydrous K2CO3. The solution Catala then filtered and used in this form in the next stage.

b. 5-PREGNAN-3,20-DIONE 20-KETAL

The above solution of 3-hydroxy-5-pregnan-20-he,20-Catala in CH2Cl2stirred with N-methylmorpholin-N-oxide (8.8 g, 75 ml) and passed through a molecular sieve (angstroms) (58 g) under nitrogen for 15 minutes. Then add perruthenate of tetrapropylammonium (400 mg) and continue stirring at room temperature for 2 hours the Obtained dark green mixture is passed through a short column with Florisil and elute CH2Cl2. The fractions containing the product (TLC), is collected and evaporated. Then the raw crystallized from a mixture of EtOAc:Hx (1:1) and obtain the target product (10.3 g) in the form of long sticks.

c. OBTAINING LITHIUM REAGENT OF 1,2-DIBROMOETHYLENE

In a 100-ml three-neck flask equipped with a bubbler for nitrogen gas, a thermometer and a dropping funnel, load 1,2-dibromoethylene (mixture of CIS/TRANS isomers 98%, Aldrich, 0,164 ml, 2 mmol, mw = 186, d = 2,246). Add dry THF (15 ml) and the solution cooled to -78oC in a bath of dry ice-acetone. Added dropwise n-butyl lithium (2.5 M in THF, 1.6 ml, 4 mmol) for 10 m is ment stage.

d. 3-ETHINYL-3-HYDROXY-5-PREGNAN-20-HE

To the above reagent solution in THF, which is kept at -78oC, is added dropwise a solution of 5 - pregnan-3,20-dione,20-Catala (180 mg, 0.5 mmol) in THF (15 ml).

In the process of adding keep the temperature below -70oC. Continue stirring at this temperature for 15 min (100% conversion according to TLC). The cooling bath is removed and conduct cooling the resulting solution by the addition of 2N HCl solution (pH 6). The solvent is removed and the residue dissolved in acetone (10 ml). After adding 2N HCl solution (4 ml) solution was stirred at room temperature for 0.5 hours. The mixture is neutralized with dilute solution of NaHCO3. A solid residue (158 mg, 93%) is filtered off, washed with water and dried. Then the crude purified by crystallization from EtOAc or a mixture of acetone-hexane to obtain the target product, So pl. 196-197oC, TLC-Rfto 0.45 (hexane:acetone 7:3).

EXAMPLE 2

THE DISODIUM SALT OF 3,20-DIHYDROXY-5-PREGNEN, BIS(HEMISUCCINATE)

Suspension 3,20-dihydroxy-5-pregnene (Steraloids: 250 mg, 0.78 mmol) in 5 ml of dry pyridine is treated with succinic anhydride (200 mg, 2.0 mmol). The mixture is heated to 100oC in General within 10 hours Additional 6 mmol of ant is UP>C) to remove solvent and then heated to 90oC (0.05 mm RT.CT.) to remove excess succinic anhydride. The residue is recrystallized from a mixture of ether/hexane, obtaining a solid residue containing mainly succinic acid. The mother liquor concentrated and subjected to column chromatography (silica gel for thin layer chromatography 95/5/0,1 CH2Cl2/MeOH/HOAc) to give a white solid product, which is recrystallized from a mixture of ether/hexane. Bis(hemisuccinate), So pl. 81-90oC, transferred to the disodium salt.

Bis(hemisuccinate) (100 mg, 0,192 mmol) dissolved in minimum volume of methanol. Add dropwise a solution of NaHCO3(2 EQ., 33 mg, 0,393 mmol) in 0.6 ml of water. After 3 h the solution is concentrated under vacuum to obtain a white solid product.

EXAMPLE 3

3-HYDROXY-3-TRIFLUOROMETHYL-5-PREGNAN-20-HE AND 3-HYDROXY-3-TRIFLUOROMETHYL-5-PREGNAN-20-HE

To a solution of 5-pregnan-3,20-dione 20-atelectasia (356 mg, 0,987 mmol) in dry THF (5 ml) was added 0.5 M to F3CSi(CH3)3(in THF; 2.5 ml, 1.25 mmol). The obtained colorless solution is cooled to 0oC and add n-Bu4NFH2O (few crystals). The cooling bath removed. Observed gas evolution (Me3SiF) and p(3:1 hexane/acetone) shows complete consumption of starting material; the new spot is moving almost with the solvent front. Next, add 1N HCl solution (about 3 ml) and the resulting biphasic mixture was stirred at room temperature overnight. TLC (3:1 hexane/acetone) shows a single spot with Rfapproximately 0.5. On the other hand, when using methylene chloride, watching two close spots, the top of which is a minor product. Add diethyl ether and water. Spend reextracting the aqueous layer with ethyl ether. The combined organic phase was washed with saturated NaHCO3and brine, dried (MgSO4), filtered and evaporated under reduced pressure, obtaining a white crystalline (foamy) precipitate, which chromatographic in a thin layer, using as eluent CH2Cl2.

Evaporation of previously obtained fractions leads to the production of 3-hydroxy-3-trifluoromethyl-5-pregnan-20-she (10 mg).

Further elution with column gives 3-hydroxy-3-trifluoromethyl-5-pregnan-20-he (200 mg), which also contains 1.5% minor product (i.e., 3-hydroxy-3-trifluoromethyl-5-pregnan-20-it), as defined by the19F-NMR and GC-mass spectroscopy. To remove this impurity attempted to conduct recrystallization in hot smhan-20-he (145 mg) can be obtained by another chromatography with CH2Cl2in a thin layer of the above mixture of 97.5:1,5, So pl. 181-183oC.

EXAMPLE 4

3-HYDROXY-3-ETHYNYL-5-PREGNAN-20-HE AND 3-HYDROXY-3-ETHYNYL-5-PREGNAN-20-HE

A solution of 5-pregnan-3,20-dione, 20-Catala (1.18 g, 3.3 mmol) in dry THF (20 ml) is treated with miniministerial (1 M solution in THF, 3.7 mmol, 3,7 ml) at -70oC. After stirring the mixture at this temperature for 5 min and then at room temperature for 2.5 h conduct the cooled mixture with a saturated solution of NH4Cl (10 ml). The solvent is removed and the residue extracted with EtOAc. The organic layer is washed with water, diluted aqueous NaHCO3, water and brine. After drying over anhydrous MgSO4the solution is filtered and evaporated, receiving raw (1.2 g). The latter is then dissolved in acetone (20 ml). After adding 1N HCl solution (10 ml) the solution was stirred at room temperature for 15 hours, the Solvent removed and the residue extracted with CH2Cl2. The organic layer is washed with water, diluted aqueous NaHCO3, water and brine. After drying over anhydrous MgSO4the solution is filtered and evaporated, receiving raw (890 mg). The latter is then dissolved in a small amount of CH2Cl2oC.

EXAMPLE 5

a. 3-HYDROXY-5-ANDROSTANE-17-ONE

To a solution of 3-hydroxy-5-androstane-17-she (6 g) and diethylazodicarboxylate (5,04 g) in THF (50 ml) is added triperoxonane acid (3.3 grams), and the mixture turns yellow. Then add triphenylphosphine (7.6 g). The reaction mixture becomes colorless and warmed up. After 5 min add sodium benzoate and then water (100 ml). The mixture is extracted with methylene chloride (CH ml) and the organic phase is dried over magnesium sulfate. The solvent is removed under vacuum and the raw hydrolyzing potassium hydroxide (10%, 10 ml) in methanol (150 ml) for 1 hour. A large part of the methanol is then removed under vacuum and the residue is distributed between methylene chloride and ammonium chloride. The product (4.7 g, 78%) was purified by chromatography (CH2Cl2:acetone = 9:1).

b. 3-HYDROXY-21-METHYL-5-PREGN-17(20)(Z)-EN

3-hydroxy-5-androstane-17-one (2 g, 6,9 mmol) are added to a Wittig reagent derived from bromide propyltrichlorosilane (13.3 g) and trebuchet potassium (3,9 g) in THF (20 ml). The reaction mixture is heated under reflux for 12 h and cooled to 25oC. Then add Airout with methylene chloride (CH ml). The organic solution is dried over potassium carbonate and the solvent is removed under vacuum. Raw purified by chromatography (acetone:methylene chloride:hexane = 1:2:7) to give 0.84 g of product (39%). It is a mixture of Z and E isomers (Z:E = 13: 1).

c. 3-TERT-BUTYLDIMETHYLSILYLOXY-21-METHYL-5-PREGN-17(20)(Z)-EN

A mixture of 5.3-hydroxy-21-methylpregna-17(20)(Z)-ene (0.84 g, of 2.66 mmol), TBDMSCI (1.2 g, 8.0 mmol) and imidazole (0,91 g, 13.3 mmol) in methylene chloride (10 ml) and DMF (30 ml) is stirred for 12 h and then add the ammonium chloride. The mixture is then extracted with methylene chloride (CH ml) and washed with brine (50 ml). The organic solution is dried over potassium carbonate and then remove the solvent until then, until there remains DMF. The residue is then dissolved in ethyl ether and washed with brine (g ml) and then dried over potassium carbonate. Chromatography with hexane results of 1.14 g of the pure product (100%).

d. 3-TERT-BUTYLDIMETHYLSILOXY-20-HYDROXY-21-METHYL-5-PREGNAN

To a solution of 3-tert-butyldimethylsiloxy-21-methyl-5-pregn-17(20)(Z)-ene (1,14 g of 2.66 mmol) in THF (30 ml) is added dropwise a THF complex-DIBORANE (1 M solution in THF, 5.3 ml) at 0oC. the Reaction mixture is heated to 25oC for 1 h Then very OSTO ml). Then add aqueous ammonium chloride and THF layer was separated using a separating funnel. The aqueous layer was extracted with diethyl ether (2x40 ml). The organic solution is dried over potassium carbonate and the solvent is removed under vacuum. The pure product is obtained column chromatography (0.52 g, 44%).

e. 3,20-DIHYDROXY-21-METHYL-5-PREGNAN

The solution, obtained by mixing HF (48%, 5 ml) and CH3CN (30 ml), added to a flask with 3-tert-butyldimethylsiloxy-20-hydroxy-21-methyl-5-pregnane (0.51 g) to give a white precipitate. The reaction mixture was stirred for 1 h and then filtered. The solid white product was washed with diethyl ether three times, which gives satisfactory analytical results (0.3 g, 79%), So pl. 227-231oC.

EXAMPLE 6

3,21-DIHYDROXY-3-TRIFLUOROMETHYL-5-19-NORPREGNA-20-HE

To a solution of 3-hydroxy-3-trifluoromethyl-5-19-norpregna-20-she (300

mg, 0.87 mmol) in toluene (15 ml) was added MeOH (1 ml) and BF3OEt2(1,4 ml, 1.13 mmol). The resulting mixture was cooled to 0oC and added Pb(OAc)4(0.54 g, to 1.21 mmol). The reaction mixture is heated to 25oC under stirring for 45 min and then add a solution of NaHCO3(saturated, 30 ml) and the mixture stirred for 1 h Then its loading is the combat solution (50 ml) and dried over K2CO3. The crude obtained after removal of solvent, dissolved in MeOH (25 ml) and add a solution of K2CO3(rich, 8 ml). The reaction mixture was stirred for 5 h and then poured into a separating funnel containing 50 ml of water. Then it is extracted with CH2Cl2(G ml). The combined extracts dried over K2CO3and the resulting crude purified by chromatography, receiving the product (160 mg) with 21-methoxycarbonyl by-product (40 mg). The product is subjected to further purification by recrystallization from 10% acetone in hexane, receiving 88 mg of pure product (28%) as a white solid product, So pl. 140-142oC.

EXAMPLE 7

3-ETHINYL-3,20-DIHYDROXY-5-PREGNEN AND 3-ETHINYL-3,20-DIHYDROXY-5-PREGNAN

To a solution of 3-ethinyl-3-hydroxy-5-pregnan-20-she (0.31 g, of 0.91 mmol) in methanol (20 ml) is added sodium borohydride (200 mg, 5.3 mmol) and stirred the mixture at 25oC for 1 h Then add a solution of ammonium chloride (50 ml) and the mixture extracted with CH2Cl2(G ml). Chromatography (EtOAc:Hex = 3:7) receive 3-ethinyl-3,20 - dihydroxy-5-pregnen (200 mg, 65%) as the main product, So pl. 221-223oC. Minor product, 3-ethinyl-3,20-dihydroxy-5-pregnen, is subjected to further OCI-DIHYDROXY-3-ATENOLOL-5-PREGNAN-20-HE AND 3-HYDROXY-3-ETHINYL-21-METHOXY-5-PREGNAN-20-HE

To a solution of 3,21-dihydroxy-3-ethinyl-5-pregnan-20-it 21-acetate (725 mg, is 1.81 mmol) in 45 ml of methanol is added at 0oC water 10% solution of K2CO3(3.75 ml). After stirring for 30 min at room temperature the mixture is again cooled to 0oC and add 2N solution is an aqueous solution of HOAc (1.8 ml). Then the reaction mixture was added to a mixture of EtOAc/water. The aqueous layer was extracted with EtOAc twice and the combined organic phases are extracted with brine, dried (Na2SO4) and concentrate. Purification by chromatography in thin layer (20 cm silica gel column with a diameter of 4 cm, elution 2 l of 20% mixture of acetone/hexane) leads to the formation of 582 mg (90%) of the diol as a white solid product, So pl. 155,5-157oC.

When receiving a large number to 5.21-diols emit less polar impurity with So pl. 176-178,5oC, which, as installed, is 3-hydroxy-3-ethinyl-21-methoxy-5-pregnan-20-one and formed, perhaps as a by-product upon receipt 21-acetate.

EXAMPLE 9

-ETHINYL-3-HYDROXY-5-PREGNAN-20-HE AND 3-ETHYNYL-3-HYDROXY-5-PREGNAN-20-HE

A solution of 5-pregnan-3,20-dione,20-Catala (720 mg, 2 mmol) in dry THF (20 ml) is treated with miniministerial (1 M solution in THF, 4 mmol, 4 ml) at -78)2
Cl2. The organic layer is washed with water, dilute NaHCO solution, water and brine. After drying over MgSO4the solution is filtered and evaporated, receiving, raw (1 g). Raw then dissolved in a small amount of CH2Cl2and poured into a column of silica gel. Elution with a mixture of toluene:acetone (95:5) gives 3-ethynyl-3 - hydroxy-5-pregnan-20-he (250 mg) as a first fraction, So pl. 163-165oC. Further elution with the same mixture solvent gives 3-ethynyl-3-hydroxy-5-pregnan-20-he (150 mg).

EXAMPLE 10

3-HYDROXY-5-TRIFLUOROMETHYL-5-19-NORPREGNA-20-HE

To a solution of 3-hydroxy-3-trifluoromethyl-5-19-norpregna-17(20)(Z)-ene (2.6 g, 7,3 mmol) in THF (80 ml) is added dropwise complex DIBORANE-THF (1 M solution in THF, 22 ml) at 25oC. the Reaction is over 1 hour and then very carefully add the solution hydrocity sodium (20%, 50 ml) at 0oC, then add the hydrogen peroxide (30%, 30 ml). Then water is added and the THF layer was separated using a separating funnel. Water words which are under vacuum. On a short column (hexane:acetone = 1:1) to obtain 1.8 g of product, which is subjected to the RCC-oxidation (RCC, 2.1 g, 9.6 mmol, sodium acetate 0.8 g, 9.6 mmol). Pure product (700 mg, 26%) purified column chromatography using ethyl acetate and hexane (15:85) as eluent, So pl. 151,5-153,0oC.

EXAMPLE 11

a. 3(R)-SPIRO-2'- OXIRAN-5,17-OXOANDROSTA

A mixture of iodide trimethylsilane (6,82 g, 31 mmol) and of potassium tert-butylate (3.5 g, 31 mmol) in THF (30 ml) is heated under reflux for 1.5 h and then cooled to 25oC. Then add 17-hydroxy-5-androstane-3-one (3 g, or 10.3 mmol) and the reaction mixture is stirred at 25oC for 2 hours and Then water is added and the mixture extracted with diethyl ether (CH ml). The extract is dried over potassium carbonate, and the solvent leads to the formation of 3 g of a fairly pure product (yield of raw sugar 96%), which is used in the next stage.

b. 3-METHYL-3-HYDROXY-5-ANDROSTANE-17-ONE

To a solution of 3(R)-Spiro-2'-oxiran - 5,17-hydroxyandrost (3 g, 9.9 mmol) in THF (50 ml) under argon add sociallyengaged (LAH) (1 M solution in THF, 10 ml) and the mixture heated under reflux for 5 min and then cooled to 25oC. Add a solution of the chloride solution is dried over potassium carbonate and then add 4 methylmorpholin-N-oxide (2.9 g, 25 mmol) and crushed molecular sieves (4 Angstrom, 10 g). The mixture was then stirred for 20 min and add perruthenate of tetrapropylammonium (200 mg). The reaction ends after 1.5 h and the reaction mixture is filtered through Florisil, which is washed with a mixture of CH2Cl2and ethyl ether (1: 2). The resulting material is purified by chromatography (30% EtOAc in hexane) to obtain the product (2.5 g, 83%).

c. 3,21-DIMETHYL-3-HYDROXY-5-PREGN-17(20)(Z)-EN

The Wittig reagent obtained by mixing a mixture of bromide n-propyltrichlorosilane (2.55 g, 6.6 mmol) and of potassium tert-butylate (0.75 g, 6.6 mmol) in THF (20 ml) for 30 min, add 3-methyl-3-hydroxy-5 - androstane-17-one (0.5 g, of 1.65 mmol) and the mixture heated under reflux for 18 hours Then the reaction mixture is cooled, adding a solution of ammonium chloride and extracted with CH2Cl2. Pure product (420 mg, 77%) was obtained by chromatography (20% EtOAc in hexane).

d. 3,20-DIHYDROXY-3,21-DIMETHYL-5-PREGNAN

To a solution of 3, 21-dimethyl-3-hydroxy-5-pregn-17(20)(Z)-s (0,42 g of 1.27 mmol) in THF (30 ml) is added dropwise complex DIBORANE-THF (1 M solution in THF, 2.6 ml) at 0oC. the Reaction is carried out by heating to 25oC for 2 h and Then very slowly add rmes stirred at 25oC for 1 h Then add aqueous ammonium chloride and THF layer was separated using a separating funnel. The aqueous layer was extracted with CH2Cl2(2x40 ml). The organic solution is dried over potassium carbonate and the solvent is removed under vacuum. The product is derived column chromatography (0.17 g, 38%). Further, it is purified by recrystallization and obtain 110 mg of the product: So pl. 200-203oC.

EXAMPLE 12

3,21-DIHYDROXY-3-TRIFLUOROMETHYL-5-PREGNAN-20-HE

The solution 3,21-dihydroxy-3 - trifluoromethyl-5-pregnan-20-she,a 21-acetate (1,36 g, a 3.06 mmol) in MeOH (75 ml) cooled to 0oC. and Then added dropwise a solution of K2CO3(10% aqueous solution of 6.45 ml of 4.67 mmol). After stirring for 1.5 h at 0oC is added dropwise a solution of acetic acid (2N water, 2.5 ml, 5.0 mmol) and the mixture is heated to room temperature. Add EtOAc, CH2Cl2and water (100 ml each) and stirred. The organic phase is separated, washed with aqueous solution of NaHCO3and NaCl, dried over MgSO4and evaporated under vacuum. The residue is purified column chromatography (hexane/EtOAc 3: 1) and obtain a white solid product (973 mg, 79%), So pl. 148-150oC.

EXAMPLE 13

3,21-DIHYDROXY-3-ETHINYL-5-PREGNAN-20-21 HE-HEMISECTION is up at 0oC solid succinic anhydride (1.2 EQ.; 180 mg of 1.80 mmol). After heating to room temperature the reaction mixture is stirred for 48 hours removing the solvent under vacuum and kneading the powder residue with hexane (2x10 ml) gives a sticky product which is dissolved as much as possible in CH2Cl2and bring in a column 2 cm in diameter, filled to 13 cm silica gel. Elution with a gradient of 5% acetone/CH2Cl2- 100% acetone gives 667 mg target acid.

To remove the pyridinium salts of the acid are dissolved in EtOAc and extracted with a 0.01 N aqueous solution of HCl (30 ml), cooled by ice. The organic layer is dried Na2SO4and concentrate. The residue 633 mg, So pl. 62-68oC, dissolved in methanol and added dropwise an aqueous solution of 116 mg (0,253 mmol) and NaHCO3. After stirring for 3.5 h, the solvent is removed under reduced pressure and the residue is ground to powder with a mixture of diethyl ether/hexane. Get a light-yellow solid product, 616 mg, solubility > 20 mg/ml

EXAMPLE 14

3-VERMEIL-3-HYDROXY-5-PREGNAN-20-HE

A mixture of n-Bu4NFH2O (7,873 g) and benzene (50 ml) is heated under reflux with trap Dean-stark during the night. The mixture, which is not yannatou temperature. To the above-mentioned concentrated solution was added a solution of 3(R)-5-pregnan-3-Spiro -2'-oxiran-20-he atelectasia (2.55 g, for 6.81 mmol) in dry benzene (15 ml + 5 ml rinse), dvukhkontsevoi a pipette. The resulting solution was concentrated on the unit molecular distillation to about 10 ml and heated under reflux for 15 minutes Because it is very difficult to apply the concentrated reaction solution on TLC plate, add dry benzene (5 ml). TLC (100:1 CH2Cl2/acetone or 3:1 hexane/acetone) showed some amount of unreacted starting material, along with two new less polar spots. The reaction mixture was again concentrated and then heated under reflux for 30 min; TLC (after dilution of the mixture benzene) showed this time almost complete consumption of the initial epoxide. As before, the mixture is concentrated and heated for some time with a reflux condenser. It should be noted that this reaction takes place only when the mixture vysokokontsentrirovannye. After cooling the mixture to room temperature (formation of light yellow precipitate) is added diethyl ether or water. Because not all of the precipitate dissolves, add CH2Cl2. Aq), dried (Na2SO4), filtered and evaporated under reduced pressure to obtain a white solid product (3.33 g), PMR spectrum which showed that he is a mixture of 4:1 3-vermeil-3-hydroxy-5-pregnan-20-he atelectasia and 3-fluoro-3-hydroxymethyl-5-pregnan-20-he atelectasia.

For hydrolysis of ketala to the above-mentioned solid product add acetone (100 ml), water (5 ml) and n-TsOH H2O (143 mg, 0,752 mmol). The pH value down by adding HCl to slightly acidic reaction. The mixture is heated for a time sufficient to obtain a clear solution, which was stirred at room temperature for 2 hours the Mixture becomes cloudy, so add CH2Cl2to obtain a clear solution. TLC showed completion of reaction. The solvents are removed under reduced pressure, obtaining a white solid product, which adds CH2Cl2and the water. The aqueous layer was re-extracted with CH2Cl2. The combined organic layers washed with saturated NaHCO3, dried (MgSO4), filtered and evaporated under reduced pressure, obtaining a white crystalline residue (2.5 g), PMR spectrum which showed that he is a mixture of 4:1 3-vermeil-3-hydroxy-5 - pregnan-20-she and 3-fluoro-3-GieMme eluent gives the target 3-vermeil-3-hydroxy-5-pregnan-20-he (1,41 g, 59%), So pl. 201-203oC.

EXAMPLE 15

3-ETHINYL-3,20-DIHYDROXY-5-PREGNEN AND 3-ETHINYL-3,20-DIHYDROXY-5-PREGNAN

To a solution of 3-ethinyl-3-hydroxy-5-pregnan-20-she (0.31 g, of 0.91 mmol) in methanol (20 ml) is added sodium borohydride (200 mg, 5.3 mmol) and stirred the mixture at 25oC for 1 h Then add a solution of ammonium chloride (50 ml) and the mixture extracted with CH2Cl2(3 times 30 ml). Chromatography (EtOAc:Hex = 3:7) receive 3-ethinyl-3,20-dihydroxy-5-pregnen (192 mg, 60%) as the main product, So pl. 195,5-197,5oC; and 3-ethinyl - 3,20-dihydroxy-5-pregnen (19 mg, 6%) as a by-product, So pl. 210-215oC (with decomp.).

EXAMPLE 16

3,20-DIHYDROXY-21-METHYL-5-PREGNAN BIS-HEMISUCCINATE

In a dry flask containing 3,20-dihydroxy-21-methyl-5-pregnen (350 mg, 1.05 mmol), add pyridine (anhydrous, 5 ml) and then added succinic anhydride (1 g, 10 mmol). The mixture is heated on an oil bath at 100oC for 20 h, and then cooled to 25oC. the Mixture is stirred with a solution of HCl (1N, 70 ml) and extracted with EtOAc (3 x 40 ml) and the extracts dried over Na2SO4. The product (0.54 g) was isolated by chromatography (7% MeOH and 0.3% HOAc in CH2Cl2) and then subjected to further purification by recrystallization from EtOAc, p the>/BR>5-3-Ethylenedioxy-androstane-17-one (6 g) are added to a Wittig reagent derived from bromide ethyltriphenylphosphonium bromide (15 g) and of potassium tert-butylate (4.5 g) in THF (15 ml). The reaction mixture is heated under reflux for 2 h and cooled to 25oC. Then add methylene chloride (80 ml) and a solution of ammonium chloride (60 ml) and the organic layer was separated using a separating funnel. The aqueous layer was extracted with methylene chloride (2 times 50 ml). The organic solution is dried over potassium carbonate and the solvent is removed under vacuum. The largest part of the oxide of phosphorus removed by washing with hexane. The resulting product (6 g) dissolved in acetone (100 ml) and add hydrochloric acid (2N, 10 ml). Raw (5.5 g) obtained by treatment with a base, followed by extraction with methylene chloride, purified by chromatography, receiving 4.5 g of product (83%). Pure stereoisomer get repeated recrystallization from hexane.

b. 3-HYDROXY-3-TRIFLUOROMETHYL-5-PREGN-17(20)(Z)-EN

To a solution of 5-pregn-17(20)(Z)-EN-3-one (950 mg, 3,17 mmol) in THF (15 ml) add triftormetilfullerenov (0.5 M solution in THF, 9.5 ml) at 0oC. the Solution gradually turns brown and the reaction over 30 minutes Add water (30 ml) and the organization is over potassium carbonate. Pure product (680 mg, 58%) was isolated column chromatography, using as eluent ethyl acetate and hexane (1:9).

EXAMPLE 18

a. 3-HYDROXY-5-PREGN-EN-20-HE

To a solution of 5-pregn-11-ene-3,20-dione (Sigma, 1.5 g, 4.77 mmol) in THF (40 ml) cautiously add three-tert-butoxyaniline lithium (1 M solution in THF, 5.7 mmol) at (-78)oC for 15 minutes the Temperature rises to 25oC for 12 h the Mixture was partitioned between ammonium chloride and diethyl ether and the resulting crude purified column chromatography (10-20% acetone in hexane, 1.3 g of product, 86%).

b. 3-HYDROXY-5-20,20 - ETHYLENEOXIDE-11-EN

To a mixture of 3-hydroxy-5-pregn-11-ene-20-she (1.3 g), ethylene glycol (8 ml) and triethylorthoformate (20 ml) is added p-toluensulfonate acid (0.1 g) and stirred the reaction mixture for 1 h Conventional treatment (sodium bicarbonate and diethyl ether), followed by removal of solvents receive the raw material, which purify by chromatography (acetone:hexane 1:4, 1.2 g, 81%).

c. 5-20,20 ETHYLENEDIOXY-5-PREGN-11-EN-3-ONE

A mixture of 3-hydroxy-20,20-Ethylenedioxy-5-pregn-11-ene (1.2 g), sodium acetate (0,53 g) and PCC (1.4 g) was stirred at 25oC for 1 h and then filtered through Florisil, elwira the P CLASS="ptx2">

d. 3-HYDROXY-3-TRIFLUOROMETHYL-5-PREGN-11-ENE-20-HE

To a solution of 5-20-ethyleneoxide-11-EN-3-one (710 mg, to 1.98 mmol) in THF (15 ml) add triftormetilfullerenov (0.5 M solution in THF, 6 ml) and then tetrabutylammonium fluoride at 0oC. the Solution gradually turns brown and the reaction is over 1 hour. Add water (30 ml) and the organic layer collected. The aqueous layer was extracted with diethyl ether (CH ml) and the organic solution is dried over potassium carbonate. Raw, obtained by removing the solvent, hydrolyzing HCl (2N solution, 5 ml) in acetone (40 ml). Pure product (554 mg, 73%) was isolated column chromatographia, using as eluent ethyl acetate and hexane (1:5). So pl. 160-162,5oC.

EXAMPLE 19

3,21-DIHYDROXY-3-ETHINYL-5 - PREGNAN-20 HE 21-ACETATE

A suspension of 3-ethinyl-3-hydroxy-5-pregnan-20-she (1,00 g of 2.92 mmol) in 35 ml of toluene is treated with 2 ml of methanol. The resulting solution is cooled in a bath of ice/water and add clean BF3-Et2O (Aldrich; 5.8 ml, 5,02 g of 35.4 mmol). Solid leads to compounds, which lead (Aldrich; 1,96 g, was 4.42 mmol) is added in several portions. Light purple solution formed first, becomes brown with continued stirring at 0oC. the Mixture is stirred PR the art 52 ml of a saturated solution of NaHCO3water and crushed ice. The resulting mixture was extracted with EtOAc (g ml). The combined organic layers extracted with saturated NaCl solution, dried (Na2SO4) and concentrate. Raw cleanse column chromatography (25 cm silica gel for thin layer in the column, with a diameter of 5, elute 3 liters of a 20% mixture of acetone/hexane) to give 749 (64%) of acetate, So pl. 196-198oC.

EXAMPLE 20

3,21-DIHYDROXY-3-VERMEIL-5-PREGNAN-20-HE, 21-ACETATE

3-Hydroxy-3-trifluoromethyl-5-pregnan-20-he (1,94 g 5,02 mmol)

dissolved in toluene (86 ml) and MeOH (5.2 ml) in an atmosphere of dry argon. Apirat triftoridov boron (10.4 ml of 84.3 mmol) is added by syringe. Then add the leads to compounds, which lead (2,89 g, 6,51 mmol). The mixture is stirred for 70 min, poured into water and extracted three times with CH2Cl2. The combined organic phases are washed with aqueous solutions of NaHCO3and NaCl, dried over MgSO4and evaporated under vacuum, obtaining a pale yellow solid product (2,18 g). This solid product was then purified column chromatography (CH2Cl2/EtOAc 150:1 and hexane/EtOAc 4:1) to give white solid product (1.54 g, 69%). So pl. 167-168,5oC.

EXAMPLE 21

3-HYDROXY-3-TRIFLUOROMETHYL-5-PREGNAN-20-HE

Reference: Krishnamurti, R; Bellew, D. R.;(3 ml) was added 0.5 M solution of F3CSi(CH3)3(in THF; 0.5 ml, 0.25 mmol). The obtained colorless solution is cooled to 0oC and add n-Bu4NFH2O (few crystals). The cooling bath removed and the mixture is heated to room temperature. In contrast, the same reaction with 5-pregnan-3,20-dione-20-ethylketone, the above reaction mixture turns yellow, and is the evolution of a gas. TLC (3:1 hexane/acetone) showed no education of any product. Then add 0.5 M solution of F3CSi(CH3)3(in THF; 0.5 ml, 0.25 mmol). The resulting mixture was stirred at room temperature for several minutes TLC shows the new spot, Rfwhich is close to 1, but still there is some amount of unreacted starting material. So add another 0.5 M solution of F3CSi(CH3)3(in THF; 0.5 ml, 0.25 mmol). The mixture is again stirred at room temperature. Not observed the presence of unreacted original ketone. Add 1N HCl solution (about 3 ml) and the resulting biphasic mixture was stirred at room temperature overnight. The spot, which is formed as a product of triptoreline, now disappears completely, and see two of the m and water. The aqueous layer was separated and extracted with diethyl ether. The combined organic layers washed with saturated NaHCO3and brine, dried (MgSO4), filtered and evaporated under reduced pressure, obtaining a white crystalline (foamy) precipitate. PMR and19F-NMR spectra indicated the precipitate shows the presence of two epimeres in the ratio of 85:15. The separation of these two epimeres is achieved by chromatography in a thin layer using a 15:1 hexane/acetone.

Evaporation of previously obtained fractions gives the minor isomer, which was not characterized, and is supposedly 3-hydroxy-3-trifluoromethyl-5-pregnan-20-one.

Further elution with column gives 3-hydroxy-3-trifluoromethyl-5-pregnan-20-he (50 mg).

EXAMPLE 22

3-HYDROXY-3-TRIFLUOROMETHYL-5-19-NORPREGNA-17(20)(Z))-EN

To a solution of 5-19-norpregna-17(20)(Z) -EN-3-one (823 mg, is 2.88 mmol) wtgf (30 ml) add triftormetilfullerenov (0.5 M solution in THF, 8.6 ml) at 0oC. the Solution gradually turns brown and the reaction over 30 minutes Add water (30 ml) and the organic layer collected. The aqueous layer was extracted with diethyl ether (CH ml) and the organic solution is dried over potassium carbonate. Pure PR is tx2">

EXAMPLE 23

3,21-HYDROXY-3-METHYL-5-PREGNEN-20-HE, 21-ACETATE

A solution of 3-hydroxy-3-methyl-5-pregnen-20-she (3.00 g, of 9.02 mmol) in dry toluene (110 ml) and methanol (6 ml) cooled in a bath of dry ice/acetone to (-75)oC. Add a clean syringe BF3-Et2O (Aldrich; 18 ml, 146 mmol), then solid leads to compounds, which lead (4,39 grams for 9.90 mmol) added in portions. The reaction does not occur at (-75)oC, and the reaction mixture is heated to (-10)oC for 4 h, the reaction Temperature rises to 0oC for a further 90 minutes According to HPLC the original substance is present as the main peak. After 1 h at 0oC is cooled to -15)oC and add 1,94 g leads to compounds, which lead, the reaction mixture is heated to 0oC. After 30 min HPLC analysis (scattering detector) gave the unadjusted ratio of product or raw material is 10:1, and after an additional 45 min, the reaction mixture is cooled to (-10)oC and add ice to the mixture of 100 ml of EtOAc, 165 ml of a saturated solution of NaHCO3and crushed ice. The aqueous layer was separated and washed with EtOAc (g ml). The combined organic layers are then washed twice with water and saturated NaCl solution. After drying over Na2SO4remove the solvent and stated vacuum and ground to powder with two portions of hexane, 100 ml The residue leaching hexane unite with the recrystallized vasectomy recrystallized a second time. The obtained solid residue contaminated 2.8% of the initial substance (according to HPLC). A third recrystallization gives 1,087 g (yield 31%) of acetate, which contains less than 2% of the source material.

EXAMPLE 24

3,21-DIHYDROXY-5-PREGNEN-20 HE 21-DISODIUM PHOSPHATE

To a solution of 21-bromo-3-hydroxy-5-pregnan-20-she (1.0 g, 2.5 mmol) in 10 ml THF at room temperature add dimensionful (2.1 g, at 7.55 mmol) and triethylamine (1,085 ml, 7.8 mmol) under stirring. The reaction mixture was then heated under reflux for 2.5 h and then cooled to room temperature. Then add dichloromethane (25 ml) and the solution transferred into a separating funnel, washed with 1N HCl solution, saturated aqueous NaHCO3, dried with MgSO4and concentrate under vacuum, getting Dienzenhofer in the form of raw oil (943 mg). The oil was dissolved in EtOH (ethanol) (50 ml) and add a few drops of sulfuric acid. In the flask is charged with 40 mg of 5% Pd/C and the mixed solution is then kept under pressure of hydrogen gas of 1 ATM at room temperature until until the reaction is completed. The catalyst was removed by filtration and the using 2N NaOH solution. Add another portion of 50 ml of MeOH and after deposition at 0oC for 1 h, filtered off solid inorganic phosphate. The solution is concentrated under vacuum and the residue is washed with hot toluene (50 ml) and then dissolved in minimum amount of MeOH. To this solution is added slowly acetone until then, until you fall a solid residue. The mixture is centrifuged, the solvent decanted and the wet solid residue is transferred into a vial and dried under vacuum, obtaining the target compound as a hygroscopic solid.

EXAMPLE 25

3,21-DIHYDROXY-3-METHYL-5-PREGNEN-20 HE 21-DISODIUM PHOSPHATE

To a solution of 21-bromo-3-hydroxy-3-methyl-5-pregnen-20-she (1.0 g, 2,43 mmol) in 10 ml THF at room temperature add dibenzodioxin (2.1 g, 7,3 mmol) and triethylamine (1,085 ml, 7,53 mmol) under stirring. The reaction mixture was then heated under reflux for 4.5 h and then cooled to room temperature. Then add dichloromethane (25 ml) and the solution transferred into a separating funnel, washed with 1N HCl solution, saturated aqueous NaHCO3, dried with MgSO4and concentrate under vacuum, getting Dienzenhofer in the form of raw oil (1,205 g). Dimensionful (790 mg, 1.3 mmol) dissolved in smedium 50 psi H2at room temperature until until the reaction is complete (by TLC). The catalyst was removed by filtration and the filtrate concentrated. The residue is dissolved in a mixture of 4:1 MeOH:water (10 ml) and titrated to pH 11 with 1 M NaOH solution. The solution is treated with acetone until you fall easily filterable precipitate, after which the mixture is cooled to 0oC and filtered to isolate the solid product raw 260 mg of the Solid product was dissolved in 20 ml of water, getting a cloudy solution, which is filtered and then concentrated, getting 220 mg of the target compound in the form of a white solid product.

EXAMPLE 26

a. 20,20-ETHYLENEDIOXY-5-PREGNAN-3-ONE

The specified connection receive with an average yield of about 90% of 3-hydroxy-5-pregnan-20-it, using the method described for 5-connections.

b. 3-ETHINYL-3-HYDROXY-5-PREGNAN-20-HE, 20-KETAL AND 3-ETHINYL-3-HYDROXY-5-PREGNAN-20-HE, 20-KETAL

In a 250-ml three-neck flask equipped with exhaust gas, a thermometer and a condenser, download complex acetylide lithium-EDA (2,75 g, 90%, 27.5 mmol). Add dry benzene (60 ml) and bubbled through a mixture of acetylene with reasonable speed. The mixture is then heated to 50-55oC in the oil bath and treated with parts of 5-pregnan-3,20-dione,20-Catala (9 to 17 o'clock The resulting suspension is cooled to 10oC and treated with a saturated solution of NaCl (5 ml). The solvent is removed and the residue is placed in water. Water-insoluble product is collected by filtration, washed with water and dried under vacuum. This raw then recrystallized from EtOAc, getting the 3-ethinyl-3-hydroxy-5-pregnan-20-he,20-ketal (at 3.35 g). The mother liquor evaporated to dryness and the residue purified column chromatography on silica gel. Elution with a mixture of toluene: acetone (92:8) to give unreacted source ketone (1.3 g) and 3-ethinyl-3-hydroxy-5-pregnan-20-he,20-ketal (1.3 g) in the second fraction. Further elution with the same mixture of solvents gives a more polar 3-ethinyl-3-hydroxy-5-pregnan-20-he,20-ketal (270 mg).

c. 3-ETHINYL-3-HYDROXY-5-PREGNAN-20-HE

3-Ethinyl-3-hydroxy-5-pregnan-20-he,20-ketal (550 mg) dissolved in a mixture of acetone (20 ml) and 2N HCl solution (10 ml) and the mixture is stirred at room temperature for 15 hours, the Solvent removed and the residue extracted with CH2Cl2. The organic layer is washed with water, diluted aqueous NaHCO3, water and brine. After drying over anhydrous MgSO4the solution is filtered and evaporated, receiving raw (414 mg). This raw then dissolved in debolsillo compound (280 mg), So pl. 175-177oC.

d. 3-ETHINYL-3-NITROXY-5-PREGNAN-20-HE

A solution of 3-ethinyl-3-hydroxy-5-pregnan-20-he,20-Catala (2.15 g) in CHCl3(45 ml) cooled to (-20)oC and treated with acetic anhydride (20 ml). Then add fuming nitric acid and the mixture is stirred at this temperature for 45 minutes After heating up (-5)oC yellow solution was poured into a mixture of 2N NaOH solution (70 ml) and water (150 ml) to give a final solution with a pH of 3-4. Then it is extracted with CHCl3, washed with water, saturated solution Panso3, brine, dried (MgSO4) and evaporated, obtaining the target product in the form of a viscous substance that is used in the next stage.

e. 3-ETHINYL-3-HYDROXY-5-PREGNAN-20-HE AND 3-ETHINYL-3-HYDROXY-5-PREGNAN-20-HE

Take raw (3 g) from the previous stage, in a mixture of THF and water (30 ml, 1:1), add AgNO3(516 mg). After stirring at room temperature for 15 h, the solvent removed and the residue extracted with CH2Cl2. The organic layer is washed with water, diluted aqueous NaHCO3, water and brine. After drying over anhydrous MgSO4the solution is filtered and evaporated, receiving, raw (2 g). This raw then dissolved in a small Conil-3-hydroxy-5-pregnan-20-he (550 mg) in the first fraction. Further

elution with the same mixture of solvents gives a more polar 3-ethinyl-3-hydroxy-5-pregnan-20-he (460 mg).

EXAMPLE 27

3,21-DIHYDROXY-3-TRIFLUOROMETHYL-5-PREGNAN-20-HE, 21-HEMISUCCINATE

In an atmosphere of dry argon 3,21-dihydroxy-3-trifluoromethyl-5-pregnan-20-he (920 mg, to 2.29 mmol), succinic anhydride (457 mg, of 4.57 mmol) and dimethylaminopyridine (14 mg) was dissolved in anhydrous pyridine. After stirring for 16 h add another 7 mg dimethylaminopyridine. After stirring for a further 3 h the solvent is removed under vacuum. The residual pyridine is removed by evaporation from toluene solution. The residue is purified column chromatography (CH2Cl2/MeOH; gradient from 100:1 to 50:1) to give a white semi-solid residue (1.1 g). This weakly contaminated solid product was dissolved in CH2Cl2, washed with water (I ml) and dried over MgSO4. The solvents are removed under vacuum, obtaining a white solid product (883 mg, 77%).

SODIUM SALT 3,21-DIHYDROXY-3-TRIFLUOROMETHYL-5-PREGNAN-20-HE,21-HEMISUCCINATE

3,21-Dihydroxy-3-trifluoromethyl-5-pregnan-20-he,21-hemisuccinate (859 mg, 1,71 mmol) dissolved in MeOH (35 ml). Then added dropwise NaHCO3(143 mg, 1.70 mmol) in water (10 ml). Add another 10 STATCOM triturated with a mixture of diethyl ether/hexane and then with hexane, getting a white solid product. Add MeOH and remove. Add heptane and removed under vacuum. The residue is triturated with hexane and dried in vacuum, obtaining a white solid product (878 mg, 98%).

EXAMPLE 28

3-ETHINYL-3-HYDROXY-5-PREGNAN-11-ENE-20-HE

To a solution of CIS/TRANS dibromoethylene (236 mg, of 1.27 mmol) in 5 ml dry THF at -78)oC is added dropwise a 1.6 M solution of BuLi in hexane (1.6 ml, 2.54 mmol) and the mixture is stirred for 30 minutes Then reduce the temperature to (-90)oC and added via cannula a solution of 5-pregn-11-ene-3,20-it in 10 ml of THF for 10 min and the reaction mixture was stirred at -90oC for another 30 min, then add 3 ml of a saturated aqueous solution of NH4Cl and the mixture is stirred at room temperature. The amount of solvent is reduced under vacuum to about 5 ml and then distribute the reaction mixture between ethyl acetate and water (25 ml each). The organic layer was washed with saturated aqueous NaCl, dried with MgSO4and concentrated in vacuo, getting raw. Chromatography in thin layer on a 6-inch column with 2 cm of silica gel, collecting fractions of 10 ml, elwira a mixture of 95:5 toluene:acetone, obtain 142 mg (65,6%) of target compound. So pl. 147-50oC.

EXAMPLE 29

3-Trifloromethyl 3-hydroxy-3 - trifluoromethyl-5-19-norpregna-20-it (see EXAMPLE 10). It was selected from the appropriate chromatographic fractions, So pl. 178-179oC.

EXAMPLE 30

3-(1-HEPTENYL)-3-HYDROXY-5-PREGNAN-20-HE

A solution of 1-heptyne (0,327 ml, 2.5 mmol) in dry THF (15 ml) is treated with n-BuLi (2.5 M in THF, 2.5 mmol, 1 ml) at -78)oC. After stirring the mixture at this temperature for 1 h, add a solution of cyclic 20-(1,2-candirectly) -pregnan-3,20-dione (360 mg, 1 mmol) in dry THF (15 ml) and the mixture is stirred at -78)oC for 1 h Then cooled by the addition of 2N solution of HCl solution (1 ml). The solvent is removed and the residue is then dissolved in acetone (10 ml). After adding 2N HCl solution (10 ml) the solution was stirred at room temperature for 1 h Add a saturated solution of NaHCO3to neutralize the acid. The solvents were removed and the residue extracted with EtOAc. The organic layer is washed with water, diluted aqueous NaHCO3, water and brine. After drying over anhydrous MgSO4the solution is filtered and evaporated, receiving raw (400 mg). Raw then dissolved in a small amount of CH2Cl2and applied to a column of silica gel. Elution with a mixture of toluene:acetone (93:7) to give 3-(1-heptenyl)-3-hydroxy-3-pregnen-20-he (260 mg) of: 3-(1-hexenyl)-3-hydroxy-5-pregnan-20-she 3-(1-octenyl)-3-hydroxy-5-pregnan-20-it, 3-(1-cyclopropylethyl)-3-hydroxy-5-pregnan-20-it, 3-(3-methylbutyl-2-EN-1-inyl)-3-hydroxy-5-pregnan-20-it, 3-(3,3-dimethylbutyl)-3-hydroxy-5-pregnan-20-it.

EXAMPLE 31

3-CYCLOPROPYLMETHYL-3-HYDROXY-5-PREGNAN-20-HE

a. 3-(5-CHLORO-1-PENTENYL)-3-HYDROXY-5-PREGNAN-20-HE-CYCLIC 20-(1,2-CANDIRECTLY)

A solution of 5-chloropentane (0.5 ml, 4.8 mmol) in dry THF (15 ml) is treated with n-BuLi (2.4 M solution in THF, 4.8 mmol, 2 ml) at (-60)oC. After stirring the mixture at a temperature (-78)oC for 0.5 h add a solution of cyclic 20-(1,2-candirectly) 5-pregnan-3,20-dione (560 mg, 1.56 mmol) in THF (15 ml), and the mixture is stirred at -78)oC for 1 h the Cooling bath removed and the mixture is cooled by a solution of NH4Cl (3 ml). The solvent is removed and the residue extracted with EtOAc. The organic layer is washed with water, diluted aqueous NaHCO3, water and brine. After drying over anhydrous MgSO4the solution is filtered and evaporated, receiving raw (660 mg). This raw use in the next stage.

b. 3-CYCLOPROPYLMETHYL-3-HYDROXY-5-PREGNAN-20-HE

The solution Diisopropylamine (0.4 ml, 3 mmol) in dry THF (15 ml) is treated with n-BuLi (2.5 M rustoleum a solution of 3-(5-chloro-1-pentyl)-3-hydroxy-5-pregnan-20-he-cyclic 20-(1,2-candirectly) (100 mg, 0.22 mmol) in THF (15 ml). The cooling bath removed and the mixture is stirred at room temperature for 0.5 h Then cooled solution of NH4Cl (3 ml). The solvent is removed, and then the residue is dissolved in acetone (40 ml). After adding 1N HCl solution (4 ml) solution was stirred at room temperature for 15 minutes Add a saturated solution of NaHCO3to neutralize the acid. The solvents were removed and the residue extracted with EtOAc. The organic layer is washed with water, diluted aqueous NaHCO3, water and brine. After drying over anhydrous MgSO4the solution is filtered and evaporated, receiving raw (120 mg). This raw then dissolved in a small amount of CH2Cl2and applied to a column of silica gel. Elution with a mixture of toluene:acetone (95:5) gives 3-(cyclopropylamino)-3-hydroxy-5-pregnan-20-he (55 mg) as a colourless solid product; So pl. 121-138oC; TLC-Rf(hexane:acetone 7:3) to 0.29.

For professionals it is obvious that the above compounds can be represented as a mixture of diastereoisomers which can be separated into individual diastereomers. Separation of diastereoisomers may be achieved by gas or liquid chromatography or the means according to the invention, as noted above, means including all isomers alone or in a mixture.

The individual isomers of the desired pharmacological activity often will prevail in one of the diastereomers. As noted, these compounds exhibit a high degree of stereospecificity. Particularly those compounds that have the greatest tool to the complex of the GABA receptor, are compounds with 3-substituted-3-hydroxypregnenolone steroid skeleton.

Compounds according to the invention and used in it, which are non-toxic, pharmaceutically acceptable natural or synthetic, directly applicable forms or proletarienne forms metabolites of progesterone, hypertension and androstane, are still unknown activity in the brain in relation to complex receptor GABAA. The present invention has the advantage that it reveals hitherto unknown mechanism and activity.

The pharmaceutical preparations according to the invention are prepared in conventional dosage forms introduction active compounds according to the invention or a mixture of such compounds in combination a non-toxic pharmaceutical nosities of activity of the subject, animal or human. Preferably the composition contains the active ingredient in an active but nontoxic amount selected in the range from about 1 mg to 500 mg of active ingredient per unit dose. This number depends on the desired specific biological activity and condition of the patient.

The desired objects of the use of compositions and methods according to the invention are in need of treatment for stress, anxiety, PMS, PND and seizures, such as caused by epilepsy, in order to facilitate or prevent attacks of anxiety, muscle tension and depression, common for patients suffering from these anomalies of the Central nervous system. Additional desirable object of the use of compositions and methods is the treatment of insomnia and getting sleeping pills. Another desirable object of using the compounds and methods is the induction of anesthesia, especially when administered intravenously.

Applied pharmaceutical carrier may be, for example, either solid, liquid media, and the media with a slow release (see, for example Remington''s Pharmaceutical Sciences, 14th Edition, 1970). Representatives of the solid carriers are allicesia cellulose, polymer hydrogels, etc., a Typical liquid media are propylene glycol, glycoluril, aqueous solutions of cyclodextrins, syrup, peanut butter and olive oil, and such emulsions. Similarly, the carrier or diluent may include any substance with delayed allocation, which are well known, such as glycerol monostearate or distearate glycerin, both by themselves and with wax, microcapsules, microspheres, liposomes and/or hydrogels.

Can be used in a wide range of pharmaceutical forms. Thus, when using a solid carrier, the drug may be raw, powdered, micronized contained in the oil, tableted, placed in a hard gelatin capsule or the capsule shell for oral administration in micronized powder or in pill form or in the form of tablets or cakes. Compounds according to the invention or used therein can also be applied in the form of suppositories for rectal administration. Connections can be added to the substance, such as cocoa butter and polyethylene glycols or other suitable non-irritating substances that are solid at room temperature, but liquid AK ampoule or as an aqueous or nonaqueous liquid suspension. Liquid dosage forms, in addition, require a pharmaceutically acceptable preservatives, etc. in Addition, because of the low doses that will be required, based on the data described here, parenteral, nasal sprays, sublingual and cheek introduction and skin patches with delayed allocation are also suitable pharmaceutical forms for topical use.

The method of obtaining activity against anxiety, anticonvulsant, mood-altering (such as antidepressant or hypnotic activity in accordance with this invention provides an introduction to the needy in such an activity of the subject compounds according to the invention is usually prepared in the composition, as described above, with a pharmaceutical carrier, in a nontoxic amount sufficient to obtain the specified activity.

During menstruation the levels of secreted metabolites vary approximately four times (Rosciszewska et al., 1986). Therefore, therapy governing symptoms includes maintaining the patient has elevated levels of metabolites of progesterone than normal in the pre-condition of patients with PMS. The levels in the patient. The number of input compounds according to the invention, as taken separately or in the form of their mixtures, calculated in such a way as to reach the level that gives the activity of a GABA receptor, equal to or exceeding the levels of metabolites of progesterone in the normal subject in premenstrual status.

The compounds may be introduced in any way that effectively delivers an active connection to the receptors GABAAsubject stimulation. The introduction can be carried out parenterally, orally, rectally, vnutrivaginalno, intracutaneous, intramuscular, sublingual, or nasal; preferred are oral, intramuscular and transdermal methods. For example, one dose in skin patch can deliver the active ingredient to a patient over a period of up to one week. However, parenteral preferred method for epilepsy.

The activity and effectiveness of plot GR

In vitro and in vivo experimental data show that the metabolites of progesterone/deoxycorticosterone and their derivatives of natural origin interact with high affinity with a new and specific site recognition on the GR complex to facilitate carry out there know, that modulation of the binding of [35S]t-butylbenzylphthalate ([35S] TBPS)is a measure of the activity and efficacy of drugs acting on the GR complex, these drugs may have potential therapeutic value in the treatment of stress, anxiety and epileptic disorders (Squires, R. F., et al., "[35S]t-Butylbicyclophophorothionate binds with high affinity to brain-specific sites coupled to gamma aminibutyric acid-A and ion recognition site", Mol. Pharmacol. , 23: 326, 1983; Lawrence J. L., et al., Benzodiazepine anticonvulsant action: gamma-aminobutiric acid-dependent modulation of the chloride ionophore", Biochem. Biophys. Res. Comm. 123:1130-1137, 1984; Wood et al., "in vitro characterization of benzodiazepine receptor agonists, antagonists, inverse agonists and agonist/antagonists", Pharmacol. Exp. Ther., 231:572-576 (1984)). We conducted a series of experiments to establish the nature of the modulation of [35S] TBPS when exposed to the compounds according to the invention. Found that these compounds interact with the new site on the GR complex, which does not overlap barbiturates, benzodiazepine, or any other previously known sites. Moreover, these compounds have high activity and effectiveness in terms of the complex GR with the strict structural requirements for the activity.

Procedures for conducting this analysis is discussed in detail in (1) Gee et al., 1987; and (2) Gee, K. W., L. J. Lawrence and H. I. Yamamura removed immediately after killing and the cerebral cortex excised on ice. The homogenate P2prepared as described previously (Gee et al., 1986). In short, the cerebral cortex gently homogenized 0,32 M solution sucrose followed by centrifugation at 1000g for 10 minutes the Supernatant is collected and centrifuged at 9000g for 20 min the precipitate P2suspended up to 10% (original wet weight/volume) suspension in 50 Mm Na/K phosphate buffer (pH 7,4) with 200 Mm NaCl to obtain a homogenate.

Aliquots hundred microliters (ml) of the homogenate P2(0.5 milligrams (mg) protein) incubated with 2 nanomolar (Nm) [35S]TBPS (TBPS-t-butylbenzylphthalate) (70-110 CI/mm; New England Nuclear, Boston, MA) in the presence or absence of the tested natural steroids or their synthetic derivatives. Test compounds dissolved in dimethyl sulfoxide (Baker Chem. Co., Phillpsbury, NJ) and added to the incubation mixture in a 5 µl aliquot. The incubation mixture was adjusted with buffer to a final volume of 1 ml Nonspecific binding observed as binding in the presence of 2 Mm TBPS. The effect and specificity of GABA (Sigma Chem. Co., St. Louis, MO) estimate, conducting all the tests in the presence of GABA + bicucullin (Sigma Chem. Co.). Incubation was conducted at 25oC for 90 min (in constant terms), satanicmajesty quantify liquid scintillation spectrophotometry. Data on the kinetics and curves dose-response for the connection/[35S]TBPS analyze the nonlinear regression method using a computerized duplicate procedure to obtain values of rate constants and IC50(concentration of the compound, which is half the maximum inhibition of the primary binding of [35S]TBPS).

Experimental data obtained through this analysis, also published by Gee et al., 1987. The data discussed in this link, shown and described in published PCT application N WO 93/03732 from 04 March 1993, In contrast, the binding site of steroids identified in this work, different from the site of the GABA/bicuculline. Shift curves dose - response induced by bicucullin, when the inhibition of binding35S]TBPS caused by alfaxalone, is not parallel. This indicates that the sites of GABA and steroids do not overlap.

The second series of experiments are conducted to demonstrate that steroids, barbiturates and benzodiazepines do not share a common binding site on the GABA receptor. The analysis is carried out in accordance with the above procedures. The obtained data on the kinetics show potentiated by 100 microns pentobarbital Na. This effect is an indication that 3-OH-5-pregnan-20-he (steroid) and pentobarbital (barbiturate) bind to independent sites.

In the third series of experiments exploring the interaction between 3-hydroxy-5-pregnan-20-one and phenobarbital Na in the potentiation of binding (3H) flunitrazepam (FLU). These experiments further confirm the claim by the claims that steroids do not share a common site of action of benzodiazepines and barbiturates. In this series of experiments reveal the influence of different concentrations of 3-hydroxy-5-pregnan-20-it is binding (3H) FLU in the presence or absence of a maximally stimulating concentration of phenobarbital Na. Because phenobarbital Na has a higher maximum efficiency than the 3-hydroxy-5-pregnan-20-he in the potentiation of binding (3H) FLU, 3-hydroxy-5-pregnan-20-it would be a full antagonist effect of phenobarbital Na simultaneously with their competitive interaction with this website. This, however, is not observed. Thus, the test data even more confirm our conclusion that some steroids, including compounds according to the invention and used to interact with the new site, other than barbiturates and what about these steroid compounds will have therapeutic profiles, other than barbiturates and benzodiazepines.

Explore various connections to establish their activity as modulators of binding (3H) FLU in vitro. These studies are carried out in accordance with the above procedures. Based on them, we set requirements structure-activity for specific interaction with a complex of GR and the order of their activity and efficiency. Table 1 shows the values of the IC50and the maximum inhibition for some compounds, including examples of compounds according to the formula of the invention. IC50is defined as the concentration of compounds inhibiting 50% of the binding of [35S]TBPS in the control experiment. This value determines the potency of the compound in vitro. The maximum inhibition is the value of the effectiveness of compounds in vitro.

As can be seen from Table 1, the compounds according to the invention and used in it are of low value IC50characterizing the concentration required to achieve 50% of maximum inhibition of binding of [35S]TBPS, while such compounds as sex steroids (estradiol and progesterone), glucocorticoids (corticosterone) and cholesterol, having high value IC50are mainly Ekwu value described herein. In order to separate this unique class of steroids from hormonal steroids, they are now called neuroactive steroids. However, sex steroids, such as progesterone, can be metabolised in the body steroids are similar to 3-hydroxy-5-pregnan-20-ONU. Thus, progesterone can be considered as a prodrug. Data TBPS correlate with the absorption ion36Cl, potentiated different 3-gidroksilirovanii steroids described Purdy R. H. et al., J. Med. Chem, 1990, 33: 1572-1581, incorporated herein by reference, and these data also correlate well with electrophysiological data obtained by measuring the activity of steroids in the potentiation of GABA-induced response in oocytes, injected GABA receptors person. This indicates that the study TBPS is approximated by measuring the ability of steroids to allosteric modulation of the activity stream Cl-.

Connection with partial activity

To the extent that the desired therapeutic activity has been patient with minimum unwanted side effects, an important aspect of this invention is the discovery of agonists with partial aktivnosti forms of these compounds. In addition, a subgroup of neuroactive steroids other than these two groups, also shows a partial activity in the study TBPS (table 1). For patients who wish to facilitate a state of anxiety or convulsions, hypnotic effect. For patients who wish to insomnia, unwanted anesthesia. It is expected that the compounds and the activity of described as agonists with partial activity, have the desired effect with minimal adverse effect.

In addition, the ability of these compounds to potentiate due to GABA increasing the flow of Cl-limited compared to the same capacity 3-hydroxy-5-pregnan-20-she's in Xenopus oocytes, injected genes receptor GABAAman.

When the rapid system of Xenopus oocytes used for testing the limited activity of some neuroactive steroids, perform the following procedure. Oocytes of Xenopus laevis (stage VI), which were "defaultusername" cleavage by collagenase (3 h at 18-23oC, 2 mg/ml collagenase A in saline Bart, not containing salts of Ca2+), inyeccion the transcripts crnc subunits 1, 1 and 1 complex of the GABA receptoroC. Induced by agonists currents recorded voltage Xenopus oocytes at a fixed potential of -60 mV, using the amplifier volt-clamp Axoclamp 2A (Axon Instuments) in the two-electrode method volt-clamp. Sensitive to voltage and current passage microelectrodes filled 3 M KCl solution, and the resistance is 1-3 Mω when measured in standard extracellular saline. Egg constantly washed with ringer's solution for frogs (120 mm NaCl; 2 mm KCl solution: 1.0 mm solution of CaCl2; 5 mm HEPES solution, pH of 7.4) at a rate of 5-7 ml/min at room temperature (17-21oC).

All medication is injected through the system transfusion. Steroids (10-2M) is prepared in the form of concentrated mother solutions in DMSO or ethanol, and then diluted with ringer's solution to a suitable concentration. The final concentrations of DMSO and ethanol with 0.2% V/V, these concentrations do not affect the resulting reaction GABA. Royal solutions all other medicines prepared in ringer's solution. The response of the membrane current slowly filtered at 100 Hz and recorded on a magnetic tape using a tape recorder FM (Racal Store 4DS) for further analysis.

The favorable effect of progesterone may be associated with different transformations of progesterone in the active metabolites of progesterone. The use of specific metabolites of progesterone in the treatment of the above-mentioned syndromes has a clear superiority over the use of progesterone due to the high activity and efficiency of metabolites and their derivatives (see Gee et al., 1987 and Table 1).

NO HORMONAL SIDE EFFECTS

It is also shown that neuroactive steroids are no hormonal side effects due to the lack of funds to progesterone and other receptors erased the us data to determine the effect of metabolites of progesterone and its derivatives and progestin R5020 binding [3H]R5020 with progesterone receptors in the uterus of rats (Gee et al., 1988).

3H-progesterone (0,15 nm) incubated with cytotam uterus of rats in the presence of test compounds. Specific binding set after incubation and compared with the control incubation without connections. The results are expressed as the percent inhibition of binding. When the binding of the compounds with progesterone receptor with high affinity, we would expect a 100% inhibition of binding at the tested concentrations. Neuroactive steroids show less than 10% inhibition of binding, indicating that they are inactive as progesterone agents.

Then studied various hormonal activity of representatives of neuroactive steroids, further study testing their potential estrogenic, mineralocorticoid and glucocorticoid activities. These activities analyzed by monitoring the ability of compounds to inhibit the binding of steroid hormones with their respective hormone receptors. The results of these experiments are presented in published PCT application WO 93/03732 published 04 March 1993, and they are fully incorporated herein by reference.

Achiev the WMD of the above steroid receptors. Thus, they do not have the expected hormonal side effects that would be caused by the binding of these steroid receptors.

ANTICONVULSANT ACTIVITY

Conduct experiments to determine the physiological correlates of the interaction neuroactive steroid and receptor GABA by assessing the ability of compounds according to the invention and used to prevent induced metrazol convulsions in mice. Mice injected with varying doses of the test compounds according to the invention for 10 min before the injection of metrazol. The start time of myoclonia (presence KLINICHESKOI activity of the forelimbs), induced metrazole determine by observing each mouse within 30 minutes From control mice metrazol (85 mg/kg) to induce convulsions in 95% of the animals. The ability of some compounds according to the invention and used to protect mice against seizures are shown in Table 2.

The ability of neuroactive steroids to protect animals from other chemical agents that cause convulsions, are shown below for 3-hydroxy-5-pregnan-20 - she 3,21-dihydroxy-5-pregnen-20-she and 3-hydroxy-3-methyl-5-pregnen-20-she. Tests on anticonvulsant activity such videopokeronline (3.15 mg/kg); strychnine (1.25 mg/kg) or vehicle (0.9% saline solution). Immediately after injection of the agent that causes convulsions, or media the mice were observed for 30 to 45 minutes Record the number of animals with tonic and/or clonic convulsions. In the test using maximal electroshock 50 mA 60 Hz down through corneal electrodes for 200 MS with the aim of inducing tonic seizure. The ability of compounds to eliminate the tonic component is defined as the end point. The total potential depression of the Central nervous system test set rotation on the rotating rod 10 min after injection of the compounds, which determine the number of mice remaining on the rotating rod (6 rpm) after 1 min in one of three experiments. ED50(the dose at which there is half of the maximum effect) set for each test and presented in Table 3 infra. The results show that neuroactive steroids in comparison with other used in the clinic anticonvulsant agents are highly effective and have profiles similar to those of clonazepam from the group of benzodiazepines. These observations suggest therapeutic suitability of these compounds as modulation CLASS="ptx2">

THE ACTION TO REMOVE THE ALARM CONDITION

The following experiments show that the metabolites of progesterone 3-OH-5-pregnan-20-he and 3-OH-5-pregnan-20-he are effective agents for the removal of the alarm condition on the four models of anxiety human use of animals, evaluating the effectiveness of the compounds for the removal of the alarm condition. Note that these two compounds illustrate the invention as an example. These synthetic derivatives in these dimensions are also given in Tables 4-6. To measure the behavioral effects of compounds against alarm condition, use four of the following models with animals: 1) test moves from light to dark; 2) elevated plus maze; 3) conflict test Geller-Seifter and 4) test Vogel.

a) Test the movement of light in the darkness

The test moves from light to dark (Crawley and Goodwin, Pharmacol. Biochem. Behav. 13: 67-70, 1980) is based on the observation that rodents in vivo strive to develop new space, but open brightly lit areas unpleasant for rodents and constrain the behavior of the conquests (Christmas and Maxwell, Neuropharmacol. 9:17-29, 1970; File J. Neurosci. Meth. 2:219-238, 1980). Many used in the clinic, aguilo moves from the bright camera in a dark chamber, whereas agents that do not have activity on removal of the alarm condition, do not show this behavioral effect (Crawley et al., Neuropharmacol., 1984, 23:531-537).

Male mice N. I. H. Swiss-Webster (Harlan, Harlan, Indianapolis, IN) weighing 15-20 g placed four in plastic cages with sawdust bedding. In the room where animals, controlled environmental conditions (22oC) when the 12-hour cycle of light/dark (600-1800 h). Feed and water given ad libitum except during testing. Experiments conducted during 700-1500 h, and the group's balance on time of day effects. Mice only imposed once the drug or the media.

The method used is a modification of previously described methods (Wieland et al., Br. Res., 1991, 565:263-268). The device contains two automated test chamber made of two parts (Model RXYZCM16, Omnitech Electronics, Columbus, OH). Open office is associated with a closed compartment through the passage 7,5x7,5 cm open office bright light using an incandescent lamp of 200 watts. The experimental room is not light. The interruption of the beam of infrared rays in each cell are automatically recorded via communication with the computer via Digiscan Analyzer (Omnitech Electronics), and the data analyzed using Integrated Lab Animal Monitoring Sy is via 10 min they are placed in the center of the lit Department. The number of movements between the illuminated and dark branches, the total activity in the illuminated chamber and the time spent in the lit chamber, measured during the 10-minute test period.

The introduction of 3-OH-5 - pregnan-20-she and 3-OH-5-pregnan-20-it is in the test move from light to dark gives considerable curve based dose - response in relation to the number of movements between light and dark chambers. Further comparisons show that the number of crossings for doses as 3-OH-5-pregnan-20-she and 3-OH-5-pregnan-20-it is significantly increased as compared with the doses tested as a control (t-test, Dunnett).

In addition, 5-OH-5-pregnan-20-he and 3-OH-5-pregnan-20-give it a significant (p < 0.01) increase in activity at 10 and 20 mg/kg compared with control groups (t-test, Dunnett). Significant differences between the two compounds at all tested doses is not marked.

b) Elevated plus maze

theoretical basis for the test with an elevated cross maze close to that of the test moves from light to dark. As described by Pellow et al. (J. Neurosci. Meth., 1985, 14:149-167), the device is elevated cross maze designed to use natural avoidance mice Otaniemi elevated cross maze allows you to make two dimensions of anxiety - determining the number of outputs in open corridors and determining the time spent in the open corridors, which are expressed in percentage of the total number of outputs and the time spent in open and closed corridors.

Male mice N. I. H. Swiss-Webster (Harlan, Harlan, Indianapolis, IN) weighing 15-20 g placed four in plastic cages with sawdust. In the room where animals, controlled environmental conditions (22oC) when the 12-hour cycle of light/dark (600-1800 h). Feed and water given ad libitum except during testing. Experiments conducted during 700-1500 h, and the group's balance on time of day effects. Mice only imposed once the drug or the media.

Used the method described previously (Lister, Psychoparmacol., 1987, 92: 180-185). The device includes two open corridor perpendicular to the two closed corridors, which are raised from the floor on 50 see Each corridor has a length of 50 cm and a wall at the closed corridors height of 40 see the Maze is completely made of black plexiglass. Over each of the open corridors placed incandescent 200 watt, to create a strong contrast between open and closed corridors.

After 10 min after injection mice N. I. H. Swiss-Webster placed in the centre of the moves in the open corridors and covered corridors, and the time spent in open corridors and enclosed corridors. All four legs must be in the corridor of measurement dependent measurements. As a consequence, the time spent in the center of the maze do not take into account, so the total time spent in open corridors, interior corridors, may not be equal to 5 minutes

Both 3-OH-5-pregnan-20-he and 3-OH-5-pregnan-20-he, in the test using the elevated cross maze show an increased number of outputs in open corridors depending on the dose. 3-OH-5-pregnan-20-it gives a significant increase in output at the dose of 20 mg/kg (p 0.05), whereas 3-OH-5-pregnan-20-it gives a significant increase in output at the dose of 5 mg/kg (p 0.05), 7.5 mg/kg (p 0.01) and 10 mg/kg (p 0,01).

In addition, 3-OH-5-pregnan-20-he and 3-OH-5-pregnan-20-give it a dose-dependent increase in time spent in open corridors. 3-OH-5-pregnan-20-it gives a significant increase in time spent in open corridors, at the dose of 10 mg/kg (p 0,01), whereas 3-OH-5-pregnan-20-it gives a significant increase in time spent in open corridors, at the dose of 7.5 mg/kg (p 0.01) and 10 mg/kg (p 0,01).

The results of these experiments are presented in published PCT application WO 93/03732, published on March 4, ia activity against the state of anxiety of the compounds according to the invention and used in the experiments with elevated cross maze in the above-described conditions.

c) Conflict test Geller-Seifter

In these tests with the simulated animal of a state of anxiety in humans using conventional conflict in rats to establish properties of drugs against anxiety. Rats are trained to press the plate for positive reinforcement when two behaviors (Geller and Seifter, Psychopharmacologia, 1960, 1:482-492). The first involves pressing on the plate with the changing speed without penalty. The second is a regime with a fixed ratio, in which each press of the record leads to positive reinforcement and punishment. Component of the penalty constitutes a conflict of the animal. Component without punishment allows you to see any answers to the depressant effects, which may have medicinal product. The answer is to remove the alarm condition would answer with a sentence without affecting the response without penalty.

In conflict experiments used male albino rats Sprague-Dawley (Charles River Labs, Wilmington, MA) weighing 250-300 g, they are on a limited diet pelleted feed Purina Lab Chow with water access at any time to maintain body weight above 85% of free feeding of young and Mature animals. Rats placed the Department who wage (punishment - reduction of penalty) and against depression 3-OH-5-pregnan-20-she and 3-OH-5 - pregnan-20-it is measured in rats conflict test Geller and Seifter (1960). In this 63-minute test hungry rats in response to pressing a lever to receive a reward sweetened milk. Mode reinforcement consists of components punishment and no punishment, changing every 15 minutes in Rats trained in the test chambers (Coulbourn instruments) with lever, hung on one wall, a small dropper, which gives the reward of 0.1 ml milk (1 part of concentrated milk Eagle: 2 parts water) and a floor of metal mesh through which produce a penalty stroke current on paws. For programming and recording use managed SKED (State Systems) minicomputer DEC PDP 11/73.

First, rats are trained to respond to continuous reinforcement and quickly learn the modes with changing 30-second, 1-minute and 2-minute intervals (VI). When the continuous reinforcement of the rat is the reward of milk after each push of the lever; when the mode VI award in the form of milk goes through infrequent and varying intervals, randomly, on average, once every 2 minutes Then in the main VI mode without punishment imposed a 3 minute conflict periods; the first ntih periods accompanied by a signal in the form of light and sound, continuous reinforcement appears again, and each time the lever is given as a reward in the form of milk and punishment in the form of short (0.25 MS) shock current on paws. The intensity of the shock in the beginning is 0.2 mA and rises daily at 0.02 mA for the gradual decrease in the number of narimani the lever up to 5 or fewer responses for conflict period. This training takes 4-6 weeks, and then note the consistently low number of responses during conflict periods and the high number of periods of absence of punishment. Induced by the drugs increased the number of punished responses taken for the activity indicator on removal of the alarm condition, while reducing the number of unpunished responses take for a response rate of depression or sedation.

Here is the summarized effects of 3-OH-5-pregnan-20-she and 3-OH-5-pregnan-20-it is in the conflict test. Both compounds lead to a significant increase in the number of punished responses, suggesting that they would be active as agents for the removal of the alarm condition. Peak effect 3-OH-5-pregnan-20-it note at a dose of 2 mg/kg, and that of 3-OH-5-PSL tests at each dose all tests with each connection are combined for comparison with the control tests with the media, using t-test for close measurements: for 3-OH-5-pregnan-20-it p < 0,02; 3-OH-5-pregnan-20-it p < 0,008).

Table 5 shows the results of determination of activities on removal of the alarm condition, the compounds according to the invention and used according to the test Geller-Seifter in the above described experimental conditions.

d) Test Vogel

The Vogel test is based on the creation of a conflict between a highly motivated behavior and disgust. For this test, a strong motivation is lust. The animal is deprived of water for 12 to 16 h to obtain motivation drinking. During the study period the animals are placed in conditions of the test, so they are accustomed to drinking bowl and to reduce fear of the new environment. After training, the animals give excess water within 2 hours during this period, the animals give normal for them, the amount of water and feed, compensating time constraints. However, this mode also gives strong motivation to drink during the testing period.

An animal that has been deprived of water for twelve or sixteen hours, placed in a cell for the test, where he is allowed to drink freely for five minutes This period is used for habituation of the animal to the environment and Polym available all the time. Twenty-four hour animal intracerebroventricular injected the drug.

After the specified delay, starting from the time of injection, the animal is again placed in a cell for testing on ten minutes, the Computer takes into account every time when the animal licks, and after every twentieth licking applies mild electrical stimulus through the language and/or paws. The electrical stimulus consists of a current of 0.6 mA with a duration of 100 MS. This procedure creates a conflict for the animal, which is reduced by the introduction used in the clinic agents for the removal of the alarm condition (e.g., valium). To get the curve of the dose - response of individual groups of animals injected with increasing doses of the test drug and conduct testing within a specified time.

Data for some representative compounds with the use of these measurements are summarized in Table 6.

PRODRUGS

Anticonvulsant activity and activity on removal of the alarm condition proletarienne form basic compounds-hydroxy-5-pregnan-20-she 3,21-dihydroxy-5-pregnen-20-she and their derivatives evaluated using the same procedure, the data of metrazol seizures count versus time after administration of the compounds (table 7).

Some additional prodrugs also test and present the results in Table 8. Modification of the basic compounds 3-hydroxy-5-pregnan-20-she 3, 21-dihydroxy-5-pregnen-20-she hydroxyl in position 3 and 21 different esters supports their biological activity and in some cases, this modification increases the protection created by the connection. Thus, the compounds according to the invention can be modified to obtain anticonvulsant activity and activity on removal of the alarm condition over a period of time with varying degrees of protection.

In contrast to the benzodiazepines, neuroactive steroids can also induce anesthesia. Their ability to induce anesthesia, it is believed, due to their ability to open a channel of chlorine ions in the absence of GABA, which is a property that does not have the benzodiazepines. Therefore, neurosteroid can act directly on the receptor in the absence of GABA, and "indirectly" in the presence of GABA. This "indirect" effect is called "modulation" of the receptor. (Lambert et al., Trends Pharmacology, Science, 1987, 8:224-227).

Compounds according to the invention and used in it can also be used when pokazani intravenous (i.v.) introduction. Animals anesthetic properties of drugs is measured by the ability of drugs to cause loss of the installation of reflex. Lost installation reflex is defined as the inability of the animal to return to normal (stand up) within 30 s after it is put on the back. Mice injected the drug intravenously in the lateral tail vein. After the introduction of the mice put on his back and watching the losing installation of reflex. Illustrating the results are shown in Table 8.

Although preferred embodiments of the invention described and illustrated, may be made of various substitutions and modifications within the scope of the invention. Accordingly, it is necessary to note that the present invention is described for illustration and does not limit it.

1. Steroid compounds of General formula

< / BR>
where

a) R4- 3'-methyl-but-3'-EN-1'-inyl or cyclopropylethyl, R5is methyl, R6- acetyl, dashed line means a single bond;

b) R4- trifluoromethyl, R5is methyl, R6- acetyl, dashed line means a single or double bond and when the link odinani, R5is methyl, R6- hydroxymethyl or hydroxyacetic, dashed line means a single bond, or a physiologically acceptable complex 21-esters, 3,20-or diesters 3,21-diesters.

2. Connection on p. 1, characterized in that it presents 3-hydroxy-3-(3'-methyl-but-3'-EN-1'-inyl)-5-pregnan-20-one.

3. Connection on p. 1, characterized in that it presents 3-hydroxy-3-trifluoromethyl-5-pregnan-20-one.

4. Connection on p. 1, characterized in that it presents 3-hydroxy-3-trifluoromethyl-5-pregn-11-EN-20-one.

5. Connection on p. 1, characterized in that it presents 3-(cyclopropyl)ethinyl-3-hydroxy-5-pregnan-20-one.

6. Steroid compound of General formula

< / BR>
where

a) R6- hydroxyacetic, R5hydrogen, when R4- trifluoromethyl and the connection represented in the form of 21-hemisuccinate sodium;

b) R6- hydroxyacetic, R5is methyl, and R4accepts the following values: hydrogen, in the case where the compound represented by bohemiacallada; ethinyl, in the case of a 21-hemisuccinate; trifluoromethyl in the case of a 21-hemifumarate sodium, methyl-21-succinate, 21-propionate;

C) R4is methyl, R5is methyl, R6- isopropyl fact, it presents 3,21-dihydroxy-3-trifluoromethyl-19-nor-5-pregnan-20-one 21-hemisuccinate sodium.

8. Pharmaceutical composition for modulation of excitability complex GRC containing an effective amount of a compound according to any one of paragraphs.1-7 and a pharmaceutically acceptable carrier or diluent.

9. Method for the treatment or prevention of stress or alarm condition of the animal, characterized in that the specified animal is administered an effective amount of a compound according to any one of paragraphs.1-7.

10. Way easier epileptic seizures in animals, characterized in that the specified animal is administered an effective amount of a compound according to any one of paragraphs.1-7.

11. A method of reducing or insomnia in animals, characterized in that the specified animal is administered an effective amount of a compound according to any one of paragraphs.1-7.

12. A method of treating mood disorders in animals, characterized in that the specified animal is administered an effective amount of a compound according to any one of paragraphs.1-7.

13. The method according to p. 12, characterized in that the mood disturbance is represented by the depression.

14. A method of treating premenstrual syndrome (PMS) or postpartum the Oia according to any one of paragraphs.1-7.

15. Method of induction of anesthesia in animals, characterized in that the specified animal is administered an effective amount of a compound according to any one of paragraphs. 1-7.

 

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