HRP20050172A2 - MICROBIOLOGICAL METHOD FOR THE PRODUCTION OF 7α-SUBSTITUTED 11α-HYDROXYSTEROIDS - Google Patents

Description

Descriptive part:

This invention relates to microbiological processes for obtaining 7α-substituted 11α-hydroxy steroids, 7α,17α-substituted 11β-halogen steroids that can be produced in this way, to processes for the production of reactive substances, and also to their use, if also to pharmaceutical preparations containing said substances. Furthermore, this invention relates to other 7α-substituted 11β-halogen steroids, especially to 7α-substituted estra-1,3,5(10)-trienes which can be obtained from 7α-substituted 11α-hydroxy steroids.

In the treatment of male menopause, as well as for the development of male sexual organs, and also for birth control, androgens, especially testosterone, are mainly used. Furthermore, said hormones also include anabolic partially active components, which, for example, stimulate muscle growth.

Male menopause is characterized by a reduction in endogenous androgen production that is associated with aging, and is treated by performing hormone replacement therapy (HRT).

In addition to the reduction in spermatogenesis, the application of LH-RH in birth control will also result in the release of LH and a decrease in testosterone and libido, which are compensated by the application of testosterone pharmaceutical reagents (D. E. Cummings et al., "Prostate-Sparing Effects of the Potent Androgen 7α- Methyl-19-Nortestosterone: A Potential Alternative to Testosterone for Androgen Replacement and Male Contraception," Journal of Clinical Endocrinology and Metabolism, Vol. 83, No. 12, pages 4212-4219 (1998)).

A combination of androgen therapy and a gestagen-active component can be used to control male fertility (see, for example, WO 01/60376 A and the documents cited therein).

In the case of treatment with testosterone, side effects have been shown to develop, particularly prostate enlargement due to an increase in the number of stromal cells and glands (BPH: benign prostatic hyperplasia). In the metabolism of testosterone, which is influenced by 5α-reductase, dihydrotestosterone (DHT) is produced, which can result, for example, in the appearance of BPH (Cummings et al., ibid.; WO 99/13883 A1). Inhibition of 5α-reductase is therefore used to treat BPH in clinical practice (finasteride).

The rapid metabolism of the androgenic steroid testosterone in the human body further results not only in the formation of unwanted DHT, but also in the fact that higher oral application doses are required in order to achieve the desired testosterone levels. Therefore, alternative forms of application are necessary, such as injections or large patches.

In order to replace testosterone in the aforementioned context, it is suggested to use 7α-methyl-19-nortestosterone (MeNT), which, on the other hand, has a higher level of biological effectiveness than testosterone, because it implies a higher level of affinity for binding to androgen receptors. On the other hand, due to steric inhibition with the 7α-methyl group, metabolism by 5α-reductase is likely to remain (Cummings et al., ibid., WO 99/13883 A1, WO 99/13812 A1, US-A-5,342,834) .

During the metabolism of testosterone, a small part of this substance also reacts by aromatizing with ring A of the steroid system, which forms estradiol, especially in the brain, liver and fat tissue. In relation to the overall action of testosterone and its metabolites, estradiol is actually responsible for sexual behavior and gonadotropin regulation. Therefore, its action is positive, just as in the case of testosterone in adult males (Cummings et al., ibid.).

However, it has been proven that the pharmacokinetics of testosterone are not satisfactory. Especially in the case of oral application, testosterone is quickly removed by excretion, so the effectiveness and duration of action of said pharmaceutical reagents is not really satisfactory. Other testosterone derivatives were subsequently synthesized for the aforementioned reasons. Such derivatives are described for example in US-A-5,952,319, in particular 7α-,11β-dimethyl derivatives of 19-nortestosterone, namely 7α,11β-dimethyl-17β-hydroxyestr-4-en-3-one, 7α,11β-dimethyl-17β -heptanoyloxyestr-4-en-3-one, 7α,11β-dimethyl-17β-[[(2-cyclopentylethyl)-carbonyl]-oxy]-estr-4-en-3-one, 7α,11β-dimethyl-17β -(phenylacetyloxy]-estr-4-en-3-one and 7α,11β-dimethyl-17β-[[(trans-4-[n-butyl]cyclohexyl)-carbonyl]-oxy]-estr-4-en- 3-he.

The aforementioned 7α,11β-dimethyl derivatives imply the aforementioned advantages, such as MeNT, implying also improved pharmacokinetics, eg efficiency and duration of action are improved compared to testosterone. Said derivatives, however, can only be produced through an extremely expensive synthesis method.

Synthesis of steroids by microbiological method is described in EP 0 900 283 B1. It is stated that ester-4-ene-3,17-dione and canrenone can be transformed with the help of microorganisms selected from the group consisting of Aspergillus nigricans, Rhizopus arrhizus and members of the genus Pestelotia into the corresponding 11α-hydroxy analog. In the introduction to the descriptive part of this invention, however, reference is made to Shibahara et al., Biochim. Biophys. Acta, 202 (1970), 172-179, where it is stated that the microbial 11α-hydroxylation reaction in steroids can be unpredictable.

The problem on which the said invention is based is therefore aimed at finding testosterone derivatives that are not sensitive to reduction with 5α-reductase, which also imply improved pharmacokinetic properties and which imply a relatively simple production process. Accordingly, a significant aspect of this invention consists in finding a process that would enable better availability of the initial products, through which the process of obtaining them would be simplified.

The problem on which this invention is based was solved through microbiological processes for the production of 7α-substituted steroids in accordance with patent claims 1, 3 and 6, 7α,17α-substituted 11β-halogen steroids in accordance with patent claim 11, through a process for the production of 7α . steroids according to claim 27, as well as 7α-substituted 11β-haloestra-1,3,5(10)-trienes according to claim 21. The preferred aspects of the claims are set out in the claims of the second order.

Definitions:

The definitions below refer to all parts of the description of this invention as well as to the patent claims attached to Diagram I:

All groups, radicals or other structural units may in each individual case be varied independently of each other in the context of the indicated areas of consideration.

All alkyl, alkylene, alkenyl, alkenylene, alkynyl and alkynylene groups may be straight chain or branched. For example, a propenyl group can be described by one of the following chemical structures: -CH=C-CH3, -CH2-C=CH2, or -C(CH3)=CH2. For example, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, n-pentyl, i-pentyl, t-pentyl, neo-pentyl, n-hexyl, 1-methyl- n-pentyl, 2-methyl-n-pentyl, 3-methyl-n-pentyl, 4-methyl-n-pentyl, 1-ethyl-n-butyl, 2-ethyl-n-butyl, etc. are represented by the term C1 - to C18-alkyl.

Alicyclic alkyl represents cycloalkyl, or cycloalkyl which is substituted with one alkyl group or several alkyl groups, which are directly connected via a cycloalkyl ring or via alkyl groups.

In the same way, alicyclic alkenyl represents cycloalkenyl, or else cycloalkenyl or cycloalkyl which is substituted with one or more alkenyl groups or else with one or more alkenyl groups and alkyl groups, or else with one or more alkyl groups, and which is directly linked via cycloalkenyl ring or via one of the alkenyl groups or optionally alkyl groups, provided that at least one double bond is contained within the alicyclic alkenyl.

On the other hand, aryl can be phenyl, but also 1-naphthyl or 2-naphthyl. In principle, aryl also includes heteroaryl, especially 2-, 3- and 4-pyridinyl, 2- and 3-furyl-, 2- and 3-thienyl, 2- and 3-pyrrolyl, 2-, 4- and 5-imidazolyl, pyridazinyl, 2-, 4- and 5-pyrimidinyl, as well as 3- and 4-pyridazinyl.

Halogen is fluorine, chlorine, bromine or iodine.

Pharmaceutically compatible addition salts are salts of the corresponding substances with inorganic and organic acids, for example with hydrochloric acid, hydrobromic acid, hydroiodic acid, acetic acid, citric acid, oxalic acid, tartaric acid and methanesulfonic acid. Esters can be formed especially with succinic acid.

The numbers in the exponent on the R symbols, for example R13, represent their positions on the steroid ring skeleton, with the C atoms in the steroid ring skeleton being numbered in accordance with IUPAC nomenclature. The numbers in the exponent on the C symbols, for example C10, indicate the position of the corresponding carbon atom on the steroid ring skeleton.

Description of the invention

Novel microbiological procedures used to obtain 7α-substituted 11α-hydroxy steroids, general formula 4,B:

[image]

which is indicated by the following parameters:

R7 represents a group P-Q, where P represents C1- to C4-alkylene and Q represents hydrogen, C1- to C4-alkyl- or C1- to C4-fluoroalkyl (alkyl which is partially or fully fluorinated), and the group P-Q is attached via P to the steroid skeleton,

R 10 represents H, CH 3 or CF 3 , and

R13 represents methyl or ethyl.

In the first variant of the procedure for obtaining said substances, a suitable 7α-substituted steroid of the general formula 3,A:

[image]

in which R7, R10 and R13 have the same meanings as given for substances of the general formula 4,B,

is hydroxylated and oxidized and by a one-step process using a microorganism selected from the group consisting of Aspergillus sp., Beauveria sp., Glomerella sp., Gnomonia sp., Haplosporella sp. and Rhizopus sp. Particularly preferred are Aspergillus awamori, Aspergillus fischeri, Aspergillus malignus, Aspergillus niger, Beauveria bassiana, Glomerella cingulata, Gnomonia cingulata, Haplosporella hesperedica and Rhizopus stolonifer, with the fact that Aspergillus awamori (CBS), Aspergillus fischeri (ATCC 1020), Aspergillus malignus (IMI 16061), Aspergillus niger (ATCC 9142), Beauveria bassiana (ATCC 7159), Glomerella cingulata (CBS 15226, CBS 23849, CBS 98069, ATCC 56596, ATCC 64682, IFO 6425), Gnomonia cingulata (CBS 15226), Haplosporella hesperedica (CBS 20837) and Rhizopus stolonifer (ATCC 15441).

As an alternative, this microbiological process can also be performed in two phases, with the hydroxylation and oxidation reactions occurring in separate steps. The progress of the reaction can be controlled through its duration: if, for example, the reaction is interrupted, after a certain time, hydroxylated but not yet oxidized species can be isolated. Both steps of said procedure can therefore be carried out separately or in fermentation:

In this context, the substance can be hydroxylated at the 11-position with the general formula 3,A in the first step of the microbiological process using the first microorganism, which is selected from the group comprising Aspergillus sp., Beauveria sp., Gibberella sp., Glomerella sp. , Gnomonia sp., Metarrhizium sp., Nigrospora sp., Rhizopus sp. and Verticillium sp., with the fact that the 7α-substituted steroid is formed with a hydroxy group at the 11α-position. This substance has the general formula C:

[image]

in which R7, R10 and R13 have the same meanings as defined above for substances of general formula 4,B. They are especially used with Aspergillus malignus, Aspergillus melleus, Aspergillus niger, Aspergillus ochraceus, Beauveria bassiana, Gibberella fujikuroi, Gibberella zeae, Glomerella cingulata, Glomerella fusaroides, Gnomonia cingulata, Metarrhizium anisopliae, Nigrospora sphaerica, Rhizopus oryzae, Rhizopus stolonifer and Verticillium dahliae. In this connection, especially Aspergillus malignus (IMI 16061), Aspergillus melleus (CBS), Aspergillus niger (ATCC 11394), Aspergillus ochraceus (NRRL 405, CBS 13252, ATCC 46504), Beauveria bassiana (ATCC 7159, IFO 5838, ATCC 13144 , IFO 4848, CBS 11025, CBS 12736), Gibberella fujikuroi (ATCC 14842), Gibberella zeae (CBS 4474), Glomerella cingulata (ATCC 10534, CBS 23849, CBS 23749, ATCC 16646, ATCC 16052, IFO 6459, IFO 6425). 6470, CBS 98069, IFO 7478, IFO 5257, ATCC 64682, ATCC 15470), Glomerella fusaroides (ATCC 9552), Gnomonia cingulata (CBS 15226), Metarrhizium anisopliae (IFO 5940), Nigrospora sphaerica (ATCC 12772), Rhizopus oryzae (ATCC 4858, ATCC 34102, ATCC 34102), Rhizopus stolonifer (ATCC 6227b, ATCC 15441) and Verticillium dahliae (ATCC 11405) are used for hydroxylation.

The intermediate product C is then oxidized in the second step of the microbiological process with the use of another microorganism selected from the group consisting of Bacillus sp., Mycobacterium sp., Nocardia sp. and Pseudomonas sp., with the formation of 7α-substituted 11α-hydroxy steroids of the general formula 4,B. Bacillus lactimorbus, Bacillus sphaericus, Mycobacterium neoaurum, Mycobacterium smegmatis, Nocardia corallina, Nocardia globerula, Nocardia minima, Nocardia restrictus, Nocardia rubropertincta, Nocardia salmonicolor and Pseudomonas testosteroni are particularly used, with particular preference being given to Bacillus lactimorbus (ATCC 245), Bacillus sphaericus (ATCC 7055), Mycobacterium neoaurum (ATCC 9626, NRRL B-3683, NRRL B-3805), Mycobacterium smegmatis (ATCC 14468), Nocardia corallina (ATCC 31338), Nocardia globerula (ATCC 9356), Nocardia minima (ATCC 19150) , Nocardia restrictus (NCIB 10027), Nocardia rubropertincta (ATCC 14352), Nocardia salmonicolor (ATCC 19149) and Pseudomonas testosteroni (ATCC 11996).

In another variant of the procedure, substances of the general formula 4,B can be produced in a microbiological reaction from 7α-substituted steroids of the general formula D:

[image]

in which R7, R10 and R13 have the same meanings as defined above for substances of general formula 4,B. This reaction is carried out using a microorganism selected from the group consisting of Aspergillus sp., Beauveria sp., Curvularia sp., Gibberella sp., Glomerella sp., Gnomonia sp., Haplosporella sp., Helicostylum sp., Nigrospora sp., Rhizopus sp. and Syncephalastrum sp., with the fact that the steroid skeleton is hydroxylated at the 11α-position and thus the 7α-substituted 11α-hydroxy steroid of the general formula 4,B is produced. Aspergillus alliaceus, Aspergillus awamori, Aspergillus fischeri, Aspergillus malignus, Aspergillus melleus, Aspergillus nidualans, Aspergillus niger, Aspergillus ochraceus, Aspergillus variecolor, Beauveria bassiana, Curvularia lunata, Gibberella zeae, Glomerella cingulata, Glomerella fusaroides, Gnomonia cingulata, Haplosporella hesperedica, Helicostylum piriformae , Nigrospora sphaerica, Rhizopus oryzae and Syncephalastrum racemosum are preferred, with Aspergillus alliaceus (ATCC 10060), Aspergillus awamori (CBS), Aspergillus fischeri (ATCC 1020), Aspergillus malignus (IMI 16061), Aspergillus melleus (CBS) being particularly frequently used. ), Aspergillus nidualans (ATCC 11267), Aspergillus niger (ATCC 9142, ATCC 11394), Aspergillus ochraceus (NRRL 405, ATCC 13252, ATCC 46504), Aspergillus variecolor (ATCC 10067), Beauveria bassiana (IFO 5838, ATCC 13144, IFO 4848 , CBS 11025, CBS 12736, ATCC 7159), Curvularia lunata (IX3), Gibberella zeae (CBS 4474), Glomerella cingulata (ATCC 10534, CBS 238 49, CBS 23749, ATCC 16646, IFO 6459, IFO 6425, IFO, 6470, ATCC 15093, ATCC 10529, IFO 5257, ATCC 56596, ATCC 64682), Glomerella fusaroides (ATCC 9552), Gnomonia cingulata (CBS 15226), Haplosporella hesperedica (CBS 20837), Helicostylum piriformae (ATCC 8992), Nigrospora sphaerica (ATCC 12772), Rhizopus oryzae (ATCC 4858) and Syncephalastrum racemosum (IFO 4827).

Processes are particularly suitable in which 7α-substituted 11α-hydroxy steroids of the general formula 4,B are obtained, in which independently R7 represents CH3 and/or R10 represents H and/or R13 represents CH3.

The procedure is carried out in the usual way. First, a sterilized nutrient solution is obtained, which is then inoculated with the culture solution. The pre-culture obtained in this way is then added to the fermenter, which is optionally coated with a suitable nutrient solution. Preferably, after the growth phase of the culture, a starting substance is added to the fermenter, which in this case means a substance of general formula 3,A or a substance of general formula D, so that the reaction can be performed in accordance with the context of consideration of this invention. After the reaction is complete, the mixture of said substances is purified in a conventional manner in order to isolate the desired 7α-substituted 11α-hydroxy steroid.

From the thus obtained substances of the general formula 4,B, other substances can also be synthesized in accordance with this invention and by methods that are also in accordance with the context of consideration of this invention. Especially 7α,17α-substituted 11β-halogen steroids of the general formula 8, 10, 12:

[image]

in which

U-V-W-X-Y-Z represents one of the ring structures C1-C2-C3-C4=C5-C10, C1-C2-C3-C4-C5=C10 or C1-C2-C3-C4-C5-C10, provided that in this case, oxo the group (=O) is attached to W (=C3), or to the ring structure C1=C2-C3=C4-C5=C6, with the fact that in this case the radical OR3 is attached to W (=C3),

R3 represents H, C1- to C4-alkyl, C1- to C4-alkanoyl or a cyclic structure C3- to C7- with an O-atom on the OR3 radical,

R7 represents the group P-Q, provided that

P represents C1- to C4-alkylene and Q represents hydrogen, C1- to C4-alkyl- or C1- to C4-fluoroalkyl (alkyl which is partially or fully fluorinated) and the group P-Q is attached via P to the steroid skeleton,

R10 may be in the α- or β-position or may represent H, CH3 or CF3, and is present only if X-Y-Z does not represent C4-C5=C10,

R11 represents halogen,

R13 represents methyl or ethyl,

R17 represents H, C1- to C18-alkyl, alicyclic element C1- to C18-alkyl, C1- to C18-alkenyl, alicyclic element C1- to C18-alkenyl, C1- to C18-alkynyl, C1- to C18-alkylaryl, C1- to C8-alkylene nitrile or a group P-Q, with the group P-Q having the meaning as previously stated and defined,

R17' represents H, C1- to C18-alkyl, alicyclic element C1- to C18-alkyl, C1- to C18-alkenyl, alicyclic element C1- to C18-alkenyl, C1- to C18-alkynyl or C1- to C18-alkylaryl , with the fact that R17' can also be attached via a keto group to the 17β-oxy group, and with the fact that R17' can also be additionally substituted with one or more NR18R19 groups or one or more SOxR20 groups, with x = 0, 1 or 2, and R18, R19 and R20 in each individual case independently of each other may have the same meaning as R17,

as well as their pharmaceutically compatible addition salts, esters and amides, represent suitable active ingredients. Said substances, which can be obtained by additional steps of the procedure from 7α-substituted 11α-hydroxy steroids of the general formula 4,B, represent valuable active ingredients with strong androgenic action without the previously mentioned side effects. Said substances are suitable for obtaining pharmaceutical reagents and especially effective contraceptive reagents and active ingredients for hormone replacement therapy (HRT).

If further R17' is substituted with the group NR18R19, it can be a methylamino, dimethylamino, ethylamino, diethylamino, cyclohexylamino, dicyclohexylamino, phenylamino, diphenylamino, benzylamino or dibenzylamino group.

Particularly suitable are 7α,17α-substituted 11β-halogen steroids of the general formula 8, 10, 12, actually substances in which U-V-W-X-Y-Z represents the ring structure C1-C2-C3-C4=C5-C10, C1-C2-C3-C4-C5=C10 or C1=C2-C3=C4-C5=C10.

In the first case, (U-V-W-X-Y-Z = C1-C2-C3-C4=C5-C10), it is about steroids of general formula 10:

[image]

In the second case (U-V-W-X-Y-Z = C1-C2-C3-C4-C5=C10), these are steroids of general formula 12:

[image]

Substances of general formulas 10 and 12 are androgenic substances.

In the third case (U-V-W-X-Y-Z = C1=C2-C3=C4-C5=C6), these are steroids of the general formula 8:

[image]

These substances are estrogens (substances with an affinity for the estrogen receptor).

In all three cases, the radicals R3, R7, R10, R11, R13, R17 and R17' have the same meanings as the corresponding radicals in the general formulas 8, 10, 12.

Independently of each other, R1 preferably represents H and/or R7 represents CH3 and/or R11 represents fluorine and/or R13 represents CH3 and/or R17 represents H, CH3, C1- to C18-alkynyl, especially ethynyl, CH2CN or CF3, and/or R17' represents H.

7α,17α-substituted 11β-halogen steroids of the general formula 8, 10, 12 which are particularly suitable in the context of consideration of this invention are:

17α-ethynyl-11β-fluoro-17β-hydroxy-7α-methylestr-4-en-3-one (Formula 10)

17α-ethynyl-11β-fluoro-17β-hydroxy-7α-methylestr-5(10)-en-3-one (Formula 12)

17α-ethynyl-11β-fluoro-7α-methylester-1,3,5(10)-triene-3,17β-diol (Formula 8).

To obtain said substances, the following production methods were adopted:

To obtain 7α,17α-substituted 11β-halogen steroids of the general formula 10, in which U-V-W-X-Y-Z represents the ring structure C1-C2-C3-C4=C5-C10, 7α-substituted 11α-hydroxy steroids of the general formula 4,B, which are obtained by a microbiological process production in accordance with the context of consideration of this invention are used as starting substances.

In the first step of the synthesis, the 7α-substituted 11α-hydroxy steroids thus obtained are converted by nucleophilic substitution with a halodehydroxylating reagent into the corresponding 7α-substituted 11β-halogen steroids 5:

[image]

[Key: Halodehydroxylierung = halodehydroxylation]

as halodehydroxylating reagents, all substances commonly used for this purpose are suitable, for example, hydrofluoric acid, hydrochloric acid, hydrobromic acid or hydroiodic acid, thionyl chloride or thionyl bromide, phosphorus pentachloride, phosphorus oxychloride, N-chlorosuccinimide, triphenylphosphine/carbon tetrachloride , HF/pyridine or diethylaminosulfur trifluoride or preferably nonaphyl fluoride/1,5-diazabicyclo[5.4.0]undecene.

Compound 10 is then produced from 5 by selective alkylation at the C17 ring skeleton (see Scheme 1). For selective alkylation, conventional alkylating reagents can be used, such as, for example, Grignard substances and organometallic substances, especially alkyllithium substances. For example, ethynylmagnesium bromide can be used as an alkylating reagent to obtain the corresponding 17α-ethynyl-17β-hydroxy-estr-4-en-3-one from estr-4-en-3,17-dione.

To obtain 7α,17α-substituted 11β-halogen steroids, in which U-V-W-X-Y-Z represents the ring structure C1-C2-C3-C4-C5=C10 and whose general formula is 12, substances of general formula 10 are used and then isomerized, in such a way that Δ4 -double bond is isomerized into Δ5(10)-double bond. In order to protect the 3-keto group, a cyclic ether at the 3-position is first formed for this purpose. Then, the Δ4-double bond is isomerized into a Δ5(10)-double bond, with the 7α,17α-substituted 11β-halogen steroid of general formula 12 being formed, while the protective group is cleaved off again.

To obtain additional 7α,17α-substituted 11β-halogen steroids of general formula 8, in which U-V-W-X-Y-Z represents C1=C2-C3=C4-C5=C10, the procedure is as follows:

First, as described above, the corresponding 11β-halogen steroid of the general formula 5 is formed from the 7α-substituted 11α-hydroxy steroid, which was obtained via microbiological hydroxylation and oxidation, and its general formula is 4,B and was obtained by halodehydroxylation nucleophilic substitution reactions.

The 7α-substituted ester-1,3,5(10)-triene of the general formula 6 is then formed by oxidation, for example with a copper(II) salt:

[image]

wherein R 3 , R 7 , R 11 and R 13 have the same meanings as above. If R 3 represents H, said substances can be synthesized directly. If some other radical is to stand instead of H in relation to R3, corresponding ethers or esters should be formed in known ways, after the 1,3,5(10)-triene ring has been formed by oxidation.

7α-substituted 11β-haloestra-1,3,5(10)-trienes of the general formula 6 as well as their pharmaceutically compatible addition salts, esters and amides also represent novel substances and are therefore covered by patent claims as intermediate products in the synthesis of 7α,17α- substituted 11β-halogen steroids of the general formula 8 also in accordance with the context of consideration of the present invention.

Particularly preferred is the 7α-substituted 11β-haloestra-1,3,5(10)-triene of general formula 6 which is 11β-fluoro-3-hydroxy-7α-methylester-1,3,5(10)-triene-17- he.

A 7α,17α-substituted 11β-halo steroid of general formula 8 in accordance with the context of consideration of the present invention can be formed from a 7α-substituted 11β-haloester-1,3,5(10)-triene of general formula 6 in the same manner as that previously described for the synthesis of the substance of general formula 10 by selective alkylation at the C17 atom of the ring skeleton.

Furthermore, 7α-substituted 11β-halogen steroids of general formula 9 can also be obtained from substances of general formula 4,B which are obtained by microbiological hydroxylation and oxidation from 7α-substituted steroids of general formula 3,A or D, and called 7α-substituted 11β- Halogen steroids also show androgenic effects:

[image]

wherein R 7 , R 11 and R 13 have the same meanings as above. A particularly preferred substance is 11β-fluoro-17β-hydroxy-7α-methylestr-4-en-3-one. Substances of general formula 9 as well as their pharmaceutically compatible addition salts, esters and amides also exhibit androgenic activity.

To obtain substances of general formula 9, estr-4-ene-3,17-dione 5 is reduced to 17β-hydroxy-estr-4-en-3-one 9, for example using boron hydride.

Furthermore, substances of the general formula 9 can be converted into the corresponding 7α-substituted 11β-haloestra-5(10)-enes:

[image]

where R7, R10, R11 and R13 have the same meaning as for formulas 8, 10, 12. For this purpose, substances of general formula 9 are isomerized by altering the Δ4-double bond into a Δ5(10)-double bond. In order to protect the 3-keto group, a cyclic ether is first formed at the 3-position. Then the Δ4-double bond is isomerized into a Δ5(10)-double bond, with the aforementioned 7α-substituted 11β-halogen steroid being formed, while the protective group is cleaved again.

Finally, the corresponding 7α-substituted 11β-haloestra-5(10)-enes can also be converted from compounds of general formula 5 by isomerization of the Δ4-double bond to the Δ5(10)-double bond:

[image]

with the proviso that R7, R10, R11 and R13 have the same meanings as in general formulas 8, 10, 12. For this purpose, substances of general formula 5 are isomerized by altering the Δ4-double bond into a Δ5(10)-double bond. In order to protect the 3-keto group, a cyclic ether is first formed at the 3-position. Then the Δ4-double bond is isomerized into a Δ5(10)-double bond, with the formation of the 7α-substituted 11β-halogen steroids mentioned above and the protective group being cleaved again.

All the aforementioned substances can also be further esterified or etherified if appropriate hydroxy groups are present at the 3- or 17β-position. For example, substance 9 can be converted into the corresponding 17β-ether or 17β-ester. The preferred substance is 11β-fluoro-17β-(4-sulfamoylbenzoxy)-7α-methylestr-4-en-3-one. As substituents on the oxy-oxygen atom at C17, basically the same radicals as mentioned for R17' are suitable.

Especially, 7α,17α-substituted 11β-halogen steroids of the general formula 8, 10, 12 are suitable for obtaining pharmaceutical reagents. This invention therefore also relates to the use of the aforementioned substances of the general formula 8, 10, 12 for the production of pharmaceutical reagents as well as pharmaceutical preparations containing at least one of the aforementioned substances of the general formula 8, 10, 12 as well as at least one pharmaceutically compatible carrier.

7α,17α-substituted 11β-halogen steroids of the general formula 10, 12, in accordance with the context of consideration of this invention, represent substances characterized by strong androgenic action, and without the aforementioned side effects, for example stimulation of the prostate (benign prostate hyperplasia, in particular, does not occur). Said substances are very easy to synthesize. It has been shown that substances in accordance with the context of consideration of this invention, general formulas 10 or 12 can be used not only for male HRT, but said substances even without the additional application of some other active ingredients, are also suitable as effective male contraceptive reagents, if previously made a good assessment at the top of the adequate lowering of the level of LH in the blood, testosterone that is produced in the body as well as FSH (follicle-stimulating hormone). This will depend on the 11β-halogen steroids in the context of the consideration of the present invention which inhibit the release of LH and FSH. LH stimulates the Leydig cells, so testosterone secretion occurs as a result. If the level of LH in the blood is low, the level of endogenous testosterone release also falls. Testosterone is needed in spermatogenesis, while FSH stimulates gametes. Sufficiently high levels of FSH and LH in the blood are therefore necessary for efficient spermatogenesis, while sufficiently high levels of LH in the blood result in the release of testosterone, which is necessary for spermatogenesis.

Since a treatment consisting exclusively of 7α,17α-substituted 11β-halogen steroids can result in effective male contraception without additional active ingredients for sterilization, the application of a pharmaceutical reagent suitable for this purpose can become significantly simpler, while the costs of use are significantly reduced.

7α,17α-substituted 11β-halogen steroids in accordance with the context of the present invention may also be used in combination with a progestogen for the purpose of controlling male fertility.

Furthermore, 7α,17α-substituted 11β-halogen steroids in accordance with the context of the present invention effectively inhibit 5α-reductase and steroid-11-hydroxylase [CYP11B (P450c11), G. Zhang, W. L. Miller, Journal of Clinical Endocrinology and Metabolism, Vol. . 81, p. 3254-3256 (1996)], in such a way that, for example, prostate stimulation is selectively avoided and said substances show significantly improved pharmacokinetic properties. Inhibition of 11-hydroxylase results in reduced deactivation of androgenic substances and their reduced excretion from the human body. As a result, the effectiveness and duration of action of said substances compared to known substances is improved, especially in the case of oral application.

Due to the aforementioned reasons, these substances are particularly suitable for use in birth control, as well as in androgen replacement therapy with a reduced tendency towards 5α-reduction, with simultaneously obtained aromatization for the purpose of forming estrogenic steroids and with a positive influence on serum lipids and on the central nervous system. System.

Androgenic action and the observation that the aforesaid side effects do not occur when the seminal vesicular test for substances of general formulas 10 and 12 is previously used is in accordance with the context of consideration of the present invention. The effectiveness of the substances of the general formula 8 in accordance with the context of consideration of the present invention was checked in the context of estrogenic action by means of the uterine growth test.

7α,17α-substituted 11β-halogen steroids of the general formula 10 or 12 in accordance with the context of consideration of this invention or pharmaceutical preparations in accordance with the context of consideration of this invention containing said substances are very suitable for treating non-sterile male patients as well as for treating all male mammals. Application for the purpose of male contraception results in the fact of temporary sterility of male patients. After the application of the active ingredients in accordance with the context of consideration of the present invention or the pharmaceutical preparations, the original state is again achieved in such a way that the male patient is no longer sterile and spermatogenesis is re-established normally. In order to keep the state of temporary sterility constant during the desired period, the application of the active ingredient or preparation should be continuous, with the same being repeated daily in short intervals or periodically in longer intervals, depending on the form of application. After a single or repeated application of the active ingredient or preparation, the non-sterile state of the male patient is not achieved immediately in the optimal case, but gradually and slowly after some time. Said period of time will depend on many factors, such as dosage, physical constitution of the patient and parallel application of other pharmaceutical reagents.

If the purpose of the application is contraception, the dosage of 7α,17α-substituted 11β-halogen steroids must be set relatively high in such a way that the levels of LH and FSH in the blood in each individual case are at most 2.5 I.E./ml (I.E.: International Units), especially at most 1.0 I.E./ml, while the level of testosterone in the blood is a maximum of 10 nmol/l, especially a maximum of 3 nmol/l.

If 7α,17α-substituted 11β-halogen steroids in accordance with the context of consideration of the present invention are to be used for HRT, and without the simultaneous need to achieve contraception, the dosage is set significantly lower. In this case, an attempt is made to achieve levels that allow the levels of LH and FSH in the blood to be significantly more than 2.5 I.E./ml, and testosterone levels to be more than 10 nmol/l.

The dosage of 7α,17α-substituted 11β-halogen steroids of the general formula 10 or 12 in accordance with the context of consideration of the present invention, which is necessary for the purpose of achieving certain levels of LH, FSH and testosterone in the blood, depends on a large number of factors and must therefore be determined specific way. First of all, it should be taken into account that the dosage is naturally related to the type of therapy itself. Substances that will be used for male contraception should be in much higher doses than in the case of using HRT. Dosage will also depend on the type of 7α,17α-substituted 11β-halogen steroid, as well as its bioavailability. The method of application is also crucial when determining the amount to be applied. Finally, the dosage will also depend in particular on the physical constitution of the patient who is to be treated, as well as on other factors, for example on the fact whether any other pharmaceutical reagents are applied in parallel.

Substances can be administered orally and parenterally, for example i.p. (intraperitoneal), i.v. (intravenous), i.m. (intramuscular) or percutaneous. The substances can also be implanted into the tissue. The amount of substance for application can fluctuate within a wide range, as long as an effective amount is applied. Depending on the condition to be treated and the method of preparation and dilution, the amount of the substance can vary widely. In humans, the daily dose can range from 0.1 to 100 mg. The daily dosage that is preferred in humans is 0.1 to 10 mg. The duration of the application will depend on the goal to be achieved.

Capsules, pills, tablets, coated tablets, creams, ointments, lotions, liquids, such as for example syrups, gels, injectable liquids, for example for i.p., i.v., i.m. or percutaneous injection, etc., are suitable forms for use during application, with the fact that individual forms for releasing the substance in accordance with the context of consideration of this invention, will imply gradual release or release of the entire amount in a short period.

Capsules, pills, tablets, coated tablets and liquids or other known oral forms for the release of pharmaceutical preparations are used for oral administration. In this case, pharmaceutical reagents can be formulated in such a way that they release the active ingredients either in a short period of time and deliver them to the body or have a so-called depot action, so that a prolonged, slow release of the active ingredient into the body is achieved. Furthermore, in addition to the 7α,17α-substituted 11β-halogen steroid, the dosage unit may contain one or more pharmaceutically compatible carriers, for example substances for adjusting the rheology of the pharmaceutical reagent, surfactants, solvents, microcapsules, microparticles, granules, diluents, binders such as starch, sugar, sorbitol and gelatin, also fillers such as silicic acid and talc, lubricants, dyes, perfumes and other substances.

In particular, the 7α,17α-substituted 11β-halogen steroids in accordance with the present invention can also be formulated in the form of a solution intended for oral application and which, in addition to the active 11β-halogen steroid, also contains the following components: pharmaceutically compatible oil and/or pharmaceutically compatible a lipophilic surfactant and/or a pharmaceutically compatible hydrophilic surfactant and/or a pharmaceutically compatible water miscible solvent. In the context of this consideration, reference is made to WO-A-97/21440.

In order to achieve better bio-availability of steroids, substances can also be formulated as cyclodextrin clathrates. For this purpose, the substances are reacted with α-β- or γ-cyclodextrin or its derivatives.

If it is intended to use creams, oils, lotions and liquids that can be applied topically, the material should be constituted in such a way that the substances in the context of consideration of this invention are dissolved in the body in sufficient quantity. Said application forms contain adjuvants, for example substances for adjusting the rheology of pharmaceutical reagents, surfactants, preservatives, solvents, diluents, substances for increasing the permeability of steroids that are considered in the context of this invention through the skin, dyes, perfumes and skin protection agents, such as are regenerators and moisturizers. Along with the steroids contemplated in the context of the present invention, other active ingredients may be included in the pharmaceutical reagent.

for parenteral application, the active ingredients may be dissolved or suspended in a physiologically compatible diluent. Oils with or without added solvents, surfactants, suspending or emulsifying reagents are often used as diluents. Examples of oils used in the context of this consideration are olive oil, peanut oil, cottonseed oil, soybean oil, castor oil, and sesame oil. For the purpose of formulating preparations for injection, any liquid carrier in which the substances considered in the context of this invention are dissolved or emulsified can be used. Said liquids often also contain substances that have the role of viscosity regulation, surfactants, preservatives, solvents, diluents and other additives, with the help of which the solution is brought to an isotonic state. Other active ingredients may also be administered together with 7α,17α-substituted 11β-halogen steroids.

The 11β-halogen steroids contemplated in the context of the present invention may be administered in the form of a depot injection or implant, for example subcutaneously, which may be formulated in such a way as to allow a delayed release of the active ingredients. In this connection, known techniques can be used, for example depots that dissolve the membrane or operate with the membrane can be used. Implants can contain inert materials, for example biodegradable polymers or synthetic silicones, such as silicone rubber. The 11β-halogen steroids contemplated in the context of the present invention may also be incorporated into, for example, a patch for the purpose of percutaneous application.

The examples given below have the purpose of explaining the invention in more detail:

A. Microbiological synthesis:

11α-hydroxy-7α-methyl-estr-4-ene-3,17-dione (Substance 4,B):

Example 1:

[image]

[Key: Mikrobiologische Hidroksilierung und Oxidation = Microbiological Hydroxylation and Oxidation]

2 l Erlenmeyer flask containing 1000 ml of nutrient solution, sterilized for 30 minutes at 121°C in an autoclave and containing 3% by weight of glucose, 1% by weight of liquid cornmeal, 0.2% by weight of NaNO3, 0.1% by weight KH2PO4, 0.2% by weight of K2HPO4, 0.05% by weight of KCl, 0.05% by weight of MgSO4.7H2O and 0.002% by weight of FeSO4.7H2O (pH 6.0) is inoculated with a culture of the genus Gnomonia cingulata (CBS 15226) and exposed to cherries for 72 hours. at 28°C in a rotary shaker at 165 rpm. Together with this culture, a 20 l fermenter is also inoculated with 19 l of sterile medium of the same final composition as described for the previous culture. Furthermore, before sterilization, the following amount of 1.0 ml of silicone oil and 1.0 ml of synperonic (oxoalcohol ethoxylate) is added in order to reduce the appearance of foam. After a growth phase of 12 hours at 0.7 bar, at a temperature of 28°C, aeration of 20 l/minute and a stirring speed of 250 rpm, a solution of 4.0 g of 17β-hydroxy-7α-methylestr-4-en-3- is added. that in 40 ml of DMF. Mixing and aeration continue. After 135 hours, the culture is counted and extracted for 12 hours with 10 l of methyl isobutyl ketone and for 5 hours with 5 l of methyl isobutyl ketone. The combined organic phases are evaporated to dryness. Silicone oil is washed with hexane. After chromatography on silica gel with a gradient consisting of hexane and ethyl acetate, 1.64 g (39%) of 11α-hydroxy-7α-methylester-4-ene-3,17-dione was isolated.

Example 2:

[image]

[Key: Mikrobiologische Hidroksilierung = Microbiological hydroxylation

Mikrobiologische Oxidation = Microbiological Oxidation]

In a 2 l Erlenmeyer vessel containing 1000 ml of nutrient solution, sterilized for 30 minutes at 121°C in an autoclave and containing 3% by weight of glucose, 1% by weight of liquid cornmeal, 0.2% by weight of NaNO3, 0.1% by weight of of KH2PO4, 0.2% by weight of K2HPO4, 0.05% by weight of KCl, 0.05% by weight of MgSO4.7H2O and 0.002% by weight of FeSO4.7H2O (pH 6.0), a culture of the genus Glomerella cingulata (IFO 6425) is inoculated, followed by cherry during 72 hours at 28°C in a rotary shaker at 165 rpm. Together with this culture, a 20 l fermenter is also inoculated with 19 l of sterile medium of the same final composition as described for the previous culture. Furthermore, before sterilization, the following amount of 1.0 ml of silicone oil and 1.0 ml of synperonic (oxoalcohol ethoxylate) is added in order to reduce the appearance of foam. After a growth phase of 12 hours at 0.7 bar, at a temperature of 28°C, aeration of 10 l/minute and a stirring speed of 350 rpm, a solution of 2.0 g of 17β-hydroxy-7α-methylestr-4-en-3- is added. that in 30 ml of DMF. Mixing and aeration continue. After 19 hours, the culture is counted and extracted during 16 hours with 20 l of methyl isobutyl ketone and during 23 hours with 20 l of methyl isobutyl ketone. The combined organic phases are evaporated to dryness. The residue is dissolved in a large amount of methanol. Silicone oil is removed by filtration and concentrated by evaporation. After chromatography on silica gel with a gradient consisting of dichloromethane and acetone, 1.55 g (73%) of 11α,17β-dihydroxy-7α-methylestr-4-en-3-one was isolated. After recrystallization from acetone/diisopropyl ether, 827 mg (39%) of a white crystal with a melting point of 163°C and [α]D = -16° (CHCl3, c = 0.501) is isolated.

In a 2 l Erlenmeyer container containing 500 ml of nutrient solution, which was sterilized for 30 minutes at 121°C in an autoclave and consists of 0.5% by weight of glucose, 0.5% by weight of bacterial-yeast extract, 0.1% by weight of peptone and 0.2% by weight of liquid cornmeal (pH 7.5), is inoculated with four cryospheres from the culture of Bacillus sphaericus (ATCC 7055) and subjected to fermentation for 24 hours at 28°C in a rotary shaker at 165 rpm.

In said pre-culture, four 2 l Erlenmeyer vessels containing 500 ml of sterile medium of the same composition as described for pre-culture are inoculated with 10% of said content. After a growth phase of 4 hours at a temperature of 28°C in a rotary shaker at 165 rpm, a solution of 50 mg of 11α,17β-dihydroxy-7α-methylestr-4-en-3-one in 2.5 ml of DMF was added to each container. Everything is subjected to cherry picking during the next 48 hours. The combined cultures are then extracted twice with 2 l of methyl isobutyl ketone. The combined organic phases are dried over sodium sulfate and then evaporated to dryness. In this case, 630 mg of oily-crystalline residue is obtained. After recrystallization from acetone/diisopropyl ether, 103 mg (49.2%) of yellowish crystals with a melting point of 189°C and [α]D = +40.4° (CHCl3, c = 0.529) were isolated (direct crystallization without prior chromatographic purification).

Example 3:

[image]

[Key: Mikrobiologische Hidroksilierung = Microbiological Hydroxylation]

In a 2 l Erlenmeyer vessel containing 500 ml of nutrient solution, which was sterilized for 30 minutes at 121°C in an autoclave and which contains 3% by weight of glucose, 1% by weight of liquid cornmeal, 0.2% by weight of NaNO3, 0.1 % by weight of KH2PO4, 0.2% by weight of K2HPO4, 0.05% by weight of KCl, 0.05% by weight of MgSO4.7H2O and 0.002% by weight of FeSO4.7H2O (pH 6.0), a culture of Aspergillus ochraceus (CBS 13252) is inoculated and subjected to cherry for 72 hours at 28°C in a rotary shaker at 165 rpm. Together with said pre-culture, a 10 l fermenter is also inoculated, and said fermenter includes an envelope of 9.5 l of sterile medium of the same final composition as described for the pre-culture. Furthermore, before the actual sterilization, the following amount of 0.5 ml of silicone oil and 0.5 ml of synperonic is added in order to reduce the formation of foam. After a growth phase of 6 hours at 0.7 bar, a temperature of 28°C, aeration of 5 l/minute and a stirring speed of 350 rpm, a solution of 1.0 g of 7α-methylester-4-ene-3,17-dione in 15 ml DMF. Mixing and aeration continue for some time. After 22 hours, the culture is analyzed and extracted twice for 4 hours with 7 l of methyl isobutyl ketone. The combined organic phases are evaporated to dryness. The residue is dissolved in a large amount of methanol. Silicone oil is removed by filtration. Concentration with evaporation follows, and after chromatography on silica gel with a gradient consisting of dichloromethane and acetone, 0.78 g (74%) of 11α-hydroxy-7α-methylestr-4-ene-3,17-dione is isolated. After recrystallization from acetone/diisopropyl ether, 311 mg (29.6%) of white crystals of melting point at 200°C and [α]D = +52° (CHCl3, c = 0.5905) are isolated.

B. Process of chemical production:

Example 4: Preparation of 11β-fluoro-17β-hydroxy-7α-methylestr-4-en-3-one:

a) 11β-fluoro-7α-methyl-estr-4-ene-3,17-dione:

11.5 ml of perfluorobutane-1-sulfonic acid fluoride is added dropwise at 0°C to a solution consisting of 13.08 g of 11α-hydroxy-7α-methyl-estr-4-ene-3,17-dione (which was obtained by microbiological synthesis in accordance with this invention [Part A]) in 250 ml of toluene and 18.2 ml of 1,8-diazabicyclo[5,4,0]undec-7-ene. After 1 hour, neutralization with 2 M hydrochloric acid followed by addition to water, extraction with ethyl acetate four times, washing with saturated sodium chloride solution, drying and concentration by evaporation in a vacuum. After the crude product is chromatographed on silica gel with a hexane/ethyl acetate gradient, 8.7 g of 11β-fluoro-7α-methyl-estr-4-ene-3,17-dione is obtained. Melting point: 101.4°C, [α]D : +135.8° (CHCl3).

b) 11β-fluoro-17β-hydroxy-7α-methylestr-4-en-3-one:

A solution of 8.7 g

11β-fluoro-7α-methyl-estr-4-ene-3,17-dione in 148 ml of tetrahydrofuran is mixed dropwise at 0°C with 29.5 ml of 1 M lithium aluminum tri-tert-butoxyhydride in tetrahydrofuran and stirred for 5.5 hour at 0°C. Dilute sulfuric acid is then added at 0°C, after which the reaction solution is added to ice water, extracted three times with ethyl acetate, washed naturally, dried over sodium sulfate, concentrated by evaporation in a vacuum and chromatographed on silica gel with hexane/ethyl acetate. 5.8 g of 11β-fluoro-17β-hydroxy-7α-methylestr-4-en-3-one with a melting point of 143-144°C is obtained. [α]D = +89.9° (CHCl3).

Example 5: Preparation of 11β-fluoro-17β-(4-sulfamoylbenzoxy)-7α-methylestr-4-en-3-one:

500 mg solution

11β-fluoro-17β-hydroxy-7α-methylestr-4-en-3-one in 7.5 ml of pyridine is mixed at room temperature with 750 mg of 4-sulfamoylbenzoic acid, 800 mg of N,N-dicyclohexylcarbodiimide, and with 125 mg of p- toluenesulfonic acid and stirred for 8.5 hours. Then everything is added to sodium bicarbonate solution, extracted four times with dichloromethane, washed in a neutral medium, dried on sodium sulfate, concentrated by evaporation in a vacuum and chromatographed on silica gel with dichloromethane/acetone. 302 mg of 11β-fluoro-17β-(4-sulfamoylbenzoxy)-7α-methylester-4-en-3-one are obtained, melting point 232°C. [α]D = +100.5° (CHCl3).

Example 6: Preparation of 17α-ethynyl-11β-fluoro-17β-hydroxy-7α-methylestr-4-en-3-one:

a) 11β-fluoro-3-methoxy-7α-methylestra-3,5-dien-17-one:

A solution of 2 g of 11β-fluoro-7α-methylester-4-ene-3,17-dione in 20 ml of 2,2-dimethoxypropane was mixed with 200 mg of pyridine tosylate for 6.5 hours at 80°C. This is followed by dilution with ethyl acetate, washing with solutions of sodium bicarbonate and sodium chloride, drying on sodium sulfate and concentration by evaporation in a vacuum. 2 g of crude 11β-fluoro-3-methoxy-7α-methylestra-3,5-dien-17-one are obtained.

b) 17α-ethynyl-11β-fluoro-17β-hydroxy-7α-methylestr-4-en-3-one:

A solution of 9.17 g of cerium(III) chloride in 60 ml of tetrahydrofuran is mixed dropwise at 0°C with 74.2 ml of ethynylmagnesium bromide solution (0.5 M in tetrahydrofuran) and stirred for 1 hour at 0°C. A solution of 2 g of crude 11β-fluoro-3-methoxy-7α-methylestra-3,5-dien-17-one in 40 ml of tetrahydrofuran is then added dropwise, followed by stirring for the next 3.5 hours at 0°C. For the purpose of refinement, a saturated solution of ammonium chloride is added, everything is then added to water, and then extracted three times with ethyl acetate, washed with semi-concentrated hydrochloric acid, sodium bicarbonate and sodium chloride solutions, followed by drying over sodium sulfate, concentration by evaporation in vacuo and chromatography on silica gel with hexane/ethyl acetate. 1.15 g of pure 17α-ethynyl-11β-fluoro-17β-hydroxy-7α-methylestr-4-en-3-one with a melting point of 218-220°C is obtained. [α]D = +19.2° (CHCl3).

Example 7: Preparation of 17α-ethynyl-11β-fluoro-17β-hydroxy-7α-methylestr-5(10)-en-3-one:

a) 3,3-ethanediyldioxy-17α-ethynyl-11β-fluoro-7α-methylestr-5(10)-en-17β-ol:

A solution of 700 mg of 17α-ethynyl-11β-fluoro-17β-hydroxy-7α-methylestr-4-en-3-one in 7 ml of dichloromethane and 4.7 ml of ethylene glycol is mixed with 2.3 ml of trimethyl orthoformate and 30 mg of p-toluenesulfonic acid hydrate. acid for 6.5 hours at room temperature. Then everything is added to sodium bicarbonate solution, extracted three times with ethyl acetate, washed in a neutral medium, dried over sodium sulfate, concentrated by evaporation in a vacuum and chromatographed on silica gel with hexane/ethyl acetate. 205 mg of 3,3-ethanediyldioxy-17α-ethynyl-11β-fluoro-7α-methylestr-5(10)-en-17β-ol are obtained.

b) 17α-ethynyl-11β-fluoro-17β-hydroxy-7α-methylestr-5(10)-en-3-one:

A solution of 205 mg of 3,3-ethanediyldioxy-17α-ethynyl-11β-fluoro-7α-methylestr-5(10)-en-17β-ol in 27 ml of methanol and 3.6 ml of water is subjected to stirring with 361 mg of oxalic acid for 24 hour at room temperature. Then everything is added to sodium bicarbonate solution, extracted three times with ethyl acetate, washed in a neutral medium, dried over sodium sulfate, concentrated by evaporation in a vacuum and chromatographed on silica gel with hexane/ethyl acetate. 95 mg of 17α-ethynyl-11β-fluoro-17β-hydroxy-7α-methylestr-5(10)-en-3-one with a melting point of 112-114°C is obtained.

Example 8: Preparation of 17α-ethynyl-11β-fluoro-7α-methylester-1,3,5(10)-triene-3,17β-diol:

a) 11β-fluoro-3-hydroxy-7α-methylestra-1,3,5(10)-trien-17-one:

A solution of 500 mg of 11β-fluoro-7α-methylester-4-ene-3,17-dione in 16.5 ml of acetonitrile is mixed with 400 mg of copper(II) bromide for 6.5 hours at 25°C. This is followed by dilution with ethyl acetate, washing with solutions of sodium bicarbonate and sodium chloride, drying on sodium sulfate, concentration by evaporation in a vacuum and chromatography on silica gel with hexane/acetone. 280 mg of pure 11β-fluoro-3-hydroxy-7α-methylestra-1,3,5(10)-trien-17-one with a melting point of 185-186°C are obtained.

b) 17α-ethynyl-11β-fluoro-7α-methylester-1,3,5(10)-triene-3,17β-diol:

A suspension of 2.03 g of cerium(III) chloride in 7.5 ml of tetrahydrofuran is mixed drop by drop at 0°C with 16.5 ml of ethynylmagnesium bromide solution (0.5 M in tetrahydrofuran) and stirred for 0.5 hours at 0°C. Then a solution of 280 mg of 11β-fluoro-3-hydroxy-7α-methylestra-1,3,5(10)-trien-17-one in 2.8 ml of tetrahydrofuran is added drop by drop and stirred for the next 3.5 hours at 0°C . For the purpose of finishing, a saturated solution of ammonium chloride is added, everything is then added to water, extracted three times with ethyl acetate, washed in a neutral medium, dried over sodium sulfate, concentrated by evaporation in a vacuum and chromatographed on silica gel with hexane/ethyl acetate.

220 mg of 17α-ethynyl-11β-fluoro-7α-methylester-1,3,5(10)-triene-3,17β-diol with a melting point of 115-117°C are obtained.

Claims (27)

1. Microbiological process for the production of 7α-substituted 11α-hydroxy steroids of general formula 4,B: [image] which is indicated by the following parameters: R7 represents the group P-Q, in which P represents C1- to C4-alkylene and Q represents hydrogen, C1- to C4-alkyl- or C1- to C4-fluoroalkyl, and the group P-Q is attached via P to the steroid skeleton, R10 can be in the α- or β-position and represents H, CH3 or CF3, i R13 represents methyl or ethyl, where the 7α-substituted steroid of general formula 3,A: [image] wherein R7, R10 and R13 have the same meanings as above, is hydroxylated and oxidized using a microorganism selected from the group consisting of Aspergillus sp., Beauveria sp., Glomerella sp., Gnomonia sp., Haplosporella sp. and Rhizopus sp. 2. The method according to patent claim 1, which is characterized by the fact that a microorganism selected from the group consisting of Aspergillus awamori, Aspergillus fischeri, Aspergillus malignus, Aspergillus niger, Beauveria bassiana, Glomerella cingulata, Gnomonia is used cingulata, Haplosporella hesperedica and Rhizopus stolonifer. 3. Microbiological process characterized by the fact that it is a process for obtaining 7α-substituted 11α-hydroxy steroids of the general formula 4,B: [image] which are indicated by the following parameters: R7 represents the group P-Q, provided that P represents C1- to C4-alkylene and Q represents hydrogen, C1- to C4-alkyl- or C1- to C4-fluoroalkyl, and the group P-Q is attached via P to the steroid skeleton, R 10 can be in the α- or β-position and represents H, CH 3 or CF 3 , and R13 represents methyl or ethyl, wherein the 7α-substituted steroid of general formula 3,A: [image] wherein R7, R10 and R13 have the same meanings as above, is hydroxylated at the 11α-position in the first step of the microbiological process using the first microorganism selected from the group that includes Aspergillus sp., Beauveria sp., Gibberella sp., Glomerella sp., Gnomonia sp., Metarrhizium sp., Nigrospora sp., Rhizopus sp. and Verticillium sp., with the formation of a 7α-substituted 11α-hydroxy steroid of the general formula C: [image] wherein R7, R10 and R13 have the same meanings as above, and The 7α-substituted 11α-hydroxy steroid of the general formula C that is produced is then oxidized in the second step of the microbiological process using another microorganism selected from the group consisting of Bacillus sp., Mycobacterium sp., Nocardia sp. and Pseudomonas sp., with the formation of a 7α-substituted steroid of the general formula 4,B. 4. The method according to patent claim 3, which is characterized in that the first microorganism is selected from the group consisting of Aspergillus malignus, Aspergillus melleus, Aspergillus niger, Aspergillus ochraceus, Beauveria bassiana, Gibberella fujikuroi, Gibberella zeae, Glomerella cingulata, Glomerella fusaroides, Gnomonia cingulata, Metarrhizium anisopliae, Nigrospora sphaerica, Rhizopus oryzae, Rhizopus stolonifer and Verticillium dahliae. 5. The method according to patent claims 3 and 4, which is characterized in that the second microorganism is selected from the group consisting of Bacillus lactimorbus, Bacillus sphaericus, Mycobacterium neoaurum, Mycobacterium smegmatis, Nocardia corallina, Nocardia globerula, Nocardia minima, Nocardia restrictus , Nocardia rubropertincta, Nocardia salmonicolor and Pseudomonas testosteroni. 6. Microbiological procedure for obtaining 7α-substituted 11α-hydroxy steroids of the general formula 4,B: [image] where R7 represents the group P-Q, provided that P represents C1- to C4-alkylene and Q represents hydrogen, C1- to C4-alkyl- or C1- to C4-fluoroalkyl, and the group P-Q is attached via P to the steroid skeleton, R 10 represents H, CH 3 or CF 3 , and R13 represents methyl or ethyl, wherein the 7α-substituted steroids of the general formula D: [image] where R7, R10 and R13 have the same meaning as above, are hydroxylated using microorganisms selected from the group consisting of Beauveria sp., Curvularia sp., Gibberella sp., Glomerella sp., Gnomonia sp., Haplosporella sp., Helicostylum sp., Nigrospora sp. and Syncephalastrum sp. 7. The method according to patent claim 6, which is characterized in that the microorganism is selected from the group consisting of Beauveria bassiana, Curvularia lunata, Gibberella zeae, Glomerella cingulata, Glomerella fusaroides, Gnomonia cingulata, Haplosporella hesperedica, Helicostylum piriformae, Nigrospora sphaerica and Syncephalastrum racemosum. 8. Microbiological process according to one of claims 1 to 7, characterized in that R7 represents CH3. 9. Microbiological process according to one of the patent claims 1 to 8, which is characterized in that R10 represents H. 10. Microbiological process according to one of claims 1 to 9, characterized in that R13 represents CH3. 11. 7α,17α-substituted 11β-halogen steroids of the general formula 8, 10, 12: [image] which are indicated by the fact that U-V-W-X-Y-Z represents one of the ring structures C1-C2-C3-C4=C5-C10, C1-C2-C3-C4-C5=C10 or C1-C2-C3-C4-C5-C10, with the fact that in this case oxo group (=O) attached to W (=C3), ring structure C1=C2-C3=C4-C5=C6, and also with the fact that in this case the radical OR3 attached to W (=C3), R3 represents H, C1- to C4-alkyl, C1- to C4-alkanoyl or cyclic C3- to C7-ether with the O-atom of the OR3-radical, R7 represents the group P-Q, in which P represents C1- to C4-alkylene and Q represents hydrogen, C1- to C4-alkyl- or C1- to C4-fluoroalkyl, and the group P-Q is attached via P to the steroid skeleton, R10 can be in the α- or β-position and represent H, CH3 or CF3, and is present only if X-Y-Z does not represent C4-C5=C10, R11 represents halogen, R13 represents methyl or ethyl, R17 represents H, C1- to C18-alkyl, alicyclic element C1- to C18-alkyl, C1- to C18-alkenyl, alicyclic C1- to C18-alkenyl, C1- to C18-alkynyl, C1- to C18-alkylaryl, C1 - that C8-alkylene nitrile or for the group P-Q, provided that the group P-Q has the above meaning, R17' represents H, C1- to C18-alkyl, alicyclic C1- to C18-alkyl, C1- to C18-alkenyl, alicyclic C1- to C18-alkenyl, C1- to C18-alkynyl or C1- to C18-alkylaryl, with provided that R17' can also be attached via a keto group to a 17β-oxy group, and provided that R17' can also be additionally substituted with one or more groups NR18R19 or with one or more groups SOxR20, provided that x = 0, 1 or 2 and R18, R19 and R20 in each individual case independently of each other can have the same meaning as R17, as well as their pharmaceutically compatible addition salts, esters and amides. 12. 7α,17α-substituted 11β-halogen steroids according to claim 11, characterized in that U-V-W-X-Y-Z represents the ring structure C1-C2-C3-C4=C5-C10 or C1-C2-C3-C4-C5=C10 or C1=C2-C3=C4-C5=C10. 13. 7α,17α-substituted 11β-halogen steroids according to one of the patent claims 11 and 12, which are characterized in that R1 represents H. 14. 7α,17α-substituted 11β-halogen steroids according to one of claims 11 and 13, which are characterized in that R7 is CH3. 15. 7α,17α-substituted 11β-halogen steroids according to one of the patent claims 11 and 14, which are characterized in that R11 is fluorine. 16. 7α,17α-substituted 11β-halogen steroids according to one of claims 11 and 15, which are characterized in that R13 represents CH3. 17. 7α,17α-substituted 11β-halogen steroids according to one of claims 11 and 16, which are characterized in that R17 represents H, CH3, C1- to C18-alkynyl, CH2CN or CF3. 18. 7α,17α-substituted 11β-halogen steroids according to one of claims 11 and 17, which are characterized in that R17 represents ethynyl. 19. 7α,17α-substituted 11β-halogen steroids according to one of claims 11 and 18, which are characterized in that R17' represents H. 20. 7α,17α-substituted 11β-halogen steroids according to one of claims 11 and 19, namely 17α-ethynyl-11β-fluoro-17β-hydroxy-7α-methylestr-4-en-3-one 17α-ethynyl-11β-fluoro-17β-hydroxy-7α-methylestr-5(10)-en-3-one 17α-Eynyl-11β-fluoro-7α-methylester-1,3,5(10)-triene-3,17β-diol. 21. 7α-substituted 11β-haloestra-1,3,5(10)-trienes of general formula 6: [image] which are indicated by the following parameters R3 represents H, C1- to C4-alkyl, C1- to C4-alkanoyl or cyclic C3- to C7-ether with O-atom of OR3-radical, R7 represents the group P-Q, in which P represents C1- to C4-alkylene and Q represents hydrogen, C1- to C4-alkyl- or C1- to C4-fluoroalkyl, and the group P-Q is attached via P to the steroid skeleton, R11 represents halogen; R13 represents methyl or ethyl, as well as their pharmaceutically compatible addition salts, esters and amides. 22. 7α-substituted 11β-haloestra-1,3,5(10)-trienes according to patent claim 21, indicated by name as 11β-fluoro-3-hydroxy-7α-methylester-1,3,5(10)- trien-17-one. 23. Process for obtaining 7α,17α-substituted 11β-halogen steroids of the general formula 10 in accordance with one of the patent claims 11 to 20, which is characterized in that U-V-W-X-Y-Z represents the ring structure C1-C2-C3-C4=C5-C10, and which includes the following steps: nucleophilic substitution on the 7α-substituted 11α-hydroxy steroid of the general formula 4,B at the 11-position with a halodehydroxylating reagent; reaction of the 7α-substituted 11β-halogen steroid which is obtained in this case with an alkylating reagent in a selective manner on the C17 atom of the ring skeleton in order to obtain a 7α,17α-substituted 11β-halogen steroid of the general formula 10. 24. Process for obtaining 7α,17α-substituted 11β-halogen steroids of the general formula 12 in accordance with one of patent claims 11 to 20, which is characterized in that U-V-W-X-Y-Z represents the ring structure C1-C2-C3-C4-C5=C10, and which includes the following procedural steps: nucleophilic substitution on the 7α-substituted 11α-hydroxy steroid of the general formula 4,B at the 11-position with a halodehydroxylating reagent, reaction of the 7α-substituted 11β-halogen steroid obtained in this case with an alkylating reagent in a selective manner on the C17 atom of the ring skeleton in order to obtain a 7α,17α-substituted 11β-halogen steroid of the general formula 10, isomerization of the 7α,17α-substituted 11β-halogen steroid of the general formula 10 in order to obtain the corresponding isomer of the general formula 12, in which U-V-W-X-Y-Z represents the ring structure C1-C2-C3-C4-C5 =C10. 25. Process for obtaining 7α,17α-substituted 11β-halogen steroids of the general formula 8 in accordance with one of the patent claims 11 to 20, in which U-V-W-X-Y-Z represents the ring structure C1=C2-C3=C4-C5=C6 with the said process implying the following steps of the procedure: nucleophilic substitution on the 7α-substituted 11α-hydroxy steroid of the general formula 4,B at the 11-position with a halodehydroxylating reagent, oxidation of the 7α-substituted 11β-halogen steroid obtained in this case for the purpose of forming the 7α-substituted ester-1,3,5(10)-triene of the general formula 6 in accordance with patent claims 17 and 18; reaction of the 7α-substituted ester-1,3,5(10)-triene of the general formula 6 with an alkylating reagent in a selective manner on the C17 atom of the ring skeleton in order to obtain the 7α,17α-substituted 11β-halogen steroid of the general formula 8. 26. The use of 7α,17α-substituted 11β-halogen steroids of the general formula 8, 10, 12 in accordance with one of the patent claims 11 to 20, which is indicated by the fact that it is aimed at obtaining pharmaceutical reagents. 27. Pharmaceutical preparations containing at least one 7α,17α-substituted 11β-halogen steroid of the general formula 8, 10, 12 in accordance with one of patent claims 11 to 20, as well as at least one pharmaceutically compatible carrier.