RU2663484C1 - METHOD OF PREPARATION 6α-METHYLHYDROCORTISONE OR ESTERS THEREOF FROM HYDROCORTISONE 21-ACETATE - Google Patents

Abstract

FIELD: chemistry.SUBSTANCE: invention relates to a process for the preparation of 6α-methylhydrocortisone (11β, 17α, 21-trihydroxy-6α-methylpregn-4-ene-3,20-dione) or esters thereof of general formula II, in which R=COCFor H; R=COCHor H, from hydrocortisone 21-acetate of formula II, through a series of conversions, comprising protection of the 11β-hydroxyl group by esterification with a trifluoroacetic acid derivative in an aprotic solvent under basic catalysis conditions to form 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxypregn-4-ene-3,20-dione of formula III, enol-esterification of a Δ-3-keto system of the resulting compound by the action of a trialkyl orthoformate in the presence of an acidic catalyst to form a 3-alkoxyester of a Δ-3-hydroxy derivative of general formula IV, condensing it with a Mannich reagent in the presence of an acid catalyst to form a mixture of 6α- and 6β-isomers of 6-(N-methyl-N-phenyl)-aminomethyl derivative of general formula V, cleavage of the resulting mixture of steroid Mannich bases on the C-N bond with the action of a mineral acid in the presence of hydrohalic acid salts to form a 6-methylene derivative of formula VI, isomerisation of the 6-exomethylene bond to the 6,7-endomethylene bond to form compound VII; catalytic hydrogenation of the 6,7-endomethylene bond to form a mixture of 6α-methyl- and 6β-methyl isomers and subsequent isomerisation of 6β-methyl isomer to a 6α-methyl derivative of formula VIII, removal of protective groups 11β- and 21-hydroxyl groups entirely or in part.II,III,IV,V,VI,VII,VIII.EFFECT: simple method for preparing a compound of formula I from hydrocortisone 21-acetate with output of desired products not less than 80 %.23 cl, 12 ex

Description

Technical field

The present invention relates to the field of organic synthesis, specifically relates to the production of steroid compounds (corticosteroids), such as 6α-methylhydrocortisone or its esters, and can be used in chemical and pharmaceutical industries, as well as in industrial biotechnology for the production of steroid drugs.

State of the art

6α-Methylhydrocortisone (CAS No. 1625-39-4) is an intermediate in the synthesis of 6α-methylprednisolone (CAS No. 83-43-2) from hydrocortisone (CAS No. 50-23-7) or 21-hydrocortisone acetate (CAS No. 50- 03-3).

Obtained in accordance with the proposed method 6α-methylhydrocortisone not only serves as an initial substrate for further conversion to 6α-methylprednisolone by chemical or biotechnological methods, but can be used to obtain its 21-esters, for example, 21-hemi-succinate 6α-methylhydrocortisone (CAS No. 119657- 85-1) and 6-methylhydrocortisone 21-acetate (CAS No. 1625-11-2).

Obtained in accordance with the proposed method, 11-trifluoroacetate 6α-methylhydrocortisone and 11-trifluoroacetate 21-acetate 6α-methylhydrocortisone can be used to obtain 17α-mono- or 17α, 21-diethylated derivatives of 6α-methylhydrocortisone, which are of interest as possible active ingredients in anti-inflammatory pharmaceutical preparations [US 4912098, 1990].

Known methods for introducing a C ⁶ -methyl substituent into a hydrocortisone or hydrocortisone 21-acetate molecule as a key step include introducing a methylene group into the C ⁶ position and then converting it to a 6α-methyl group.

The introduction of a C ⁶ -methylene group is carried out by various methods using formaldehyde or its derivatives as methyleneating agents. However, all methods include the obligatory preliminary enolization of the α, β-unsaturated ketone of ring A, which is necessary for polarization of the double bond system with the formation of a nucleophilic C ⁶ carbon atom, followed by replacement of the hydrogen atom at C ⁶ with a formyl or methylene group and differ in that in some In some cases, the formed intermediate 3,5-dienol is isolated in the form of ether [D. Burn et al., Tetrahedron, 1965, 21 (6), 1619-1624] or 3,5-dienamine [F. Schneider et al., Helv. Chim. Acta, 1973, 56 (7), 2396-2404], in other cases enolization is carried out in situ without isolation of the intermediate product.

The most attractive are the methods of so-called direct condensation with formaldehyde or its derivatives. This is the direct γ-methyleneation method proposed by Annen et al. [US 4322349, 1982; Synthesis, 1982, No. 1, p. 34-40; EP 0100874, 1984; EP 0149222, 1985], and a condensation method with a Mannich reagent [R. Bohlmann et al, DE 4121484, 1993].

The direct γ-methyleneation method involves the interaction of a steroid with formaldehyde derivatives (such as dimethyl acetal, formaldehyde diethyl acetal or methoxymethyl acetate) in chlorine-containing solvents at the boiling point in the presence of an excess of phosphorus oxychloride and sodium acetate. However, this method is not suitable for the methyleneation of compounds containing an unprotected 11β-hydroxyl group. So, with direct γ-methyleneation of hydrocortisone 21-acetate, the yield of the 6-methylene derivative is only 18% [Synthesis, 1982, No. 1, p. 34-40, Table 1, compound 6n].

It is also known that the enolesterification of the Δ ⁴ -3-ketosystem of the steroid ring A catalyzed by p-toluenesulfonic acid (p-TSC) in the presence of an unprotected 11β-hydroxyl group may be accompanied by an undesired reaction of its elimination with the formation of the Δ ^{9 (11)} bond [Zhao Q . et al. Synth. Commun., 1993, 23 (10), 1473-1478]. Therefore, to preserve the 11β-hydroxyl group, its protection is necessary.

It is known that in order to protect the 11β-hydroxy group, it is usually esterified to a trihaloacetate (in particular trifluoroacetate) or trimethylsilyl ether. These protections are considered the most acceptable, since they are not only easily formed, but also easily removed. The use of the method of esterification with lower acid anhydrides to protect the 11β-hydroxy group is possible, but limited due to problems arising from their subsequent removal by chemical methods of solvolysis.

Methods of using the trifluoroacetate protection of the 11β-hydroxyl group of the hydrocortisone (II) 21-acetate molecule are known. This is the method described in the patent [US 4330541, 1982, example 1], which consists in the fact that the esterification reaction of 5 g of hydrocortisone 21-acetate is carried out in pyridine by trifluoroacetic anhydride at a temperature of minus 15 ° C. Then the mixture was poured into an aqueous solution of sodium chloride, the filtered precipitate was dissolved in dichloromethane, the organic solution was washed with water, dried with sodium sulfate and evaporated to dryness. Obtain 5.1 g (82.4%) of the residue containing 11-trifluoroacetate 21-hydrocortisone (III) acetate. However, this compound was not characterized, since the residue containing it without purification and isolation of the crystalline product was used by the authors of the patent in the next stage of chemical synthesis.

Other patents [DE 1112511, 1961 (Example 16) and DE 1125422, 1962 (Example 15)] describe a process for the preparation of 11-trifluoroacetate 21-hydrocortisone (III) acetate from 21-hydrocortisone (II) acetate by trifluoroacetic anhydride in absolute dioxane. After 18 hours at room temperature, the reaction mixture was poured onto ice water. Then the product is extracted, the extract is washed with water, evaporated to dryness in vacuo. After crystallization of the residue in ether and chromatography on alumina, 11-trifluoroacetate of 21-hydrocortisone (III) acetate is obtained, the yield of which is not indicated. Product characterized: mp 206-207 ° C, [α] _D + 165.7 ° (hlf); λ _max 238-240 mμ. There is no spectral characteristic.

The preparation of methyl 3,5-dienol ether of 11-trifluoroacetate 21-hydrocortisone acetate (IVa) has not been described in literature.

The preparation of ethyl 3,5-dienol ether of 11-trifluoroacetate 21-hydrocortisone acetate (IVb) is described in patents [DE 1112511, 1961 (example 16) and DE 1125422, 1962 (example 15)] and consists in the reaction of compound III with ethylorthoformate in dioxane in the presence of p-TSC at room temperature. After completion of the reaction, the mixture was poured into water and extracted with dichloromethane. The extract is washed with water, dried over sodium sulfate and evaporated to dryness. An oily residue containing compound IVb, the yield of which is not indicated, is characterized by: [α] _D ± 0 ° (hlf); λ _max 240-242 mμ, log ε 4.15. Compound IVb was not isolated in crystalline form by the authors. Spectral characteristic, absent, melting point not determined.

6-Aminomethyl derivatives of the general formula V are novel compounds not described in the literature.

The 6-Methylene derivative of 11-trifluoroacetate 21-hydrocortisone VI acetate is a new compound, not described in the literature.

6,7-Dehydro-6-methyl-derivative of 11-trifluoroacetate 21-hydrocortisone VII acetate VII is a new compound, not described in the literature.

Compound VIII - 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6α-methylpregn-4-en-3,20-dione (11-trifluoroacetate 21-acetate 6α-methylhydrocortisone) is known. However, literature describes its preparation from other compounds. Thus, a method is known for producing 11-trifluoroacetate 21-acetate 6α-methylhydrocortisone from 21-acetate 6α-methylhydrocortisone by trifluoroacetylation of its 11β-hydroxyl group [US 4912098, 1990], according to which 5 g of 21-acetate 6α-methylhydrocortisone is treated with trifluoroacetic anhydride in solution at a temperature of minus 15 ° C. Then the reaction mass is poured onto an aqueous solution of sodium chloride, the filtered precipitate is dissolved in dichloromethane, the organic solution is washed with water, dried with sodium sulfate and evaporated to dryness. Obtain 5.1 g (83%) of the residue containing 11-trifluoroacetate 21-acetate 6α-methylhydrocortisone (VIII), which without crystallization is used in the next stage of chemical synthesis.

A method is also known for the preparation of compound VIII starting from 21-acetoxy-11β, 17α-bis- (trifluoroacetyloxy) -6-methylenehydrocortisone [RU 2297423, 2007], according to which compound VIII is obtained by reduction of the 6-methylene group of 21-acetoxy-11β, 17α -bis- (trifluoroacetyloxy) -6-methylenehydrocortisone into 6-methyl boiling for 24 hours in ethanol in the presence of cyclohexene and a PdO / C catalyst. In this case, the trifluoroacetyl protection of the 17α-hydroxyl group is simultaneously removed.

A known method of producing 11-trifluoroacetate 6α-methylhydrocortisone (IX) described in the patent [RU 2337918, 2008, example 17]. However, compound IX in option 1 of example 17 of the aforementioned patent was obtained by another method, namely from 11β-trifluoroacetyloxy-17α, 20; 20,21-bismethylenedioxy-6α-methylpregn-4-en-3,20-dione by removing the bismethylene protection of the dioxiaacetone side chain in an environment of acetic acid at a temperature of 0-2 ° C by the action of concentrated hydrochloric acid (20 ml of acetic acid and 75 ml of hydrochloric acid are used per 5 g of steroid). In Embodiment 2 of Example 17 of the same patent, Compound IX was prepared from 6-methylenehydrocortisone 11-trifluoroacetate by catalytic hydrogenation (Pd / C 5.4%) in the presence of cyclohexene in a mixture of ethanol and acetic acid.

A known method for producing 21-acetate of 6α-methylhydrocortisone [DE 4121484, 1993, example 2], which consists in introducing a methylene group at position C ^{6 of the} molecule of 21-acetate of hydrocortisone and subsequent catalytic reduction of the C ⁶ -methylene group to 6α-methyl. According to the description of this method, 5 g of hydrocortisone (II) 21-acetate are suspended in a mixture of 50 ml of tetrahydrofuran, 15 ml of triethylorthoformate and 15 ml of ethanol, 500 mg of pyridinium tosylate are added and stirred for 1 h at a temperature of 40 ° C in a bath. Then add 5.5 ml of N-methylaniline and 6.5 ml of a 40% formaldehyde solution and mix for at least an hour at a temperature of 40 ° C. After this, the reaction mass is treated with ice water containing tartaric acid and sodium chloride, extracted with dichloromethane, washed with a solution of sodium bicarbonate and sodium chloride, dried with sodium sulfate and evaporated to dryness in vacuo. The resulting residue (5.8 g) was chromatographed over 600 g of silica gel with hexane-ethyl acetate to give 2.1 g (40.8%) of 6-methylene derivative of 21-hydrocortisone acetate, which was then placed in a mixture of 9 ml of cyclohexene and 124 ml ethanol, add 170 mg of catalyst 10% Pd / C and boil for 8 hours. After cooling, the catalyst is filtered off and washed with ethanol, 2 drops of hydrochloric acid are added to the filtrate and concentrated in vacuo to 1/3 of the original volume, the residue is diluted with ice brine, precipitate filtered, washed with water until neutral and, dried in vacuum at 70 ° C. After recrystallization of the crude product from a mixture of acetone and hexane, 1.78 g of pure 21-acetoxy-11β, 17α-dihydroxy-6a-methylpregn-4-en-3,20-dione are obtained with so pl. 208-209 ° C and [α] _D = + 137 ° (in pyridine). Thus, the yield of 6α-methylhydrocortisone 21-acetate by the method of [DE 4121484, 1993] from hydrocortisone (II) 21-acetate is 34.4%.

The disadvantages of the process described in [DE 4121484, 1993] are:

- the use for the purification of 21-acetate 6-methylene-hydrocortisone chromatography, which significantly complicates the process.

- low yield of 6-methylene-hydrocortisone 21-acetate product (40.8%) at the methyleneation stage, which can be explained, firstly, by the presence of adverse reactions involving an unprotected hydroxyl group at C ¹¹ , and, secondly, by a low regional orientation Mannich steroid base cleavage reactions. It is likely that under the described conditions the deamination reaction (CN bond cleavage) is accompanied by a side deaminomethylation process (CC bond cleavage) with the regeneration of hydrocortisone 21-acetate. Therefore, chromatographic purification of the product becomes necessary.

The closest to the essence of the proposed is the method [RU 2297423, 2007, examples 3 (option 2) and 4], which consists in introducing a methylene group at position C ^{6 of the} 11β, 17α, 21-trialkanoyloxy-pregn-4-en-3 molecule , 20-dione and subsequent catalytic reduction of the C ⁶ -methylene group to the 6α-methyl. According to the description of this method (example 3, option 2), 10 ml of ethanol, 10 ml of triethyl orthoformate, 130 mg of p-TSC are added to a solution of 10 g of 11,17,21-hydrocortisone triacetate in 75 ml of tetrahydrofuran and heated to 40 ° C. The reaction mass was incubated for 1 h and an additional 130 mg of p-TSC was added. After stirring for another 1 h at a temperature of 40 ° C, 4.3 ml of N-methylaniline and 3-6 ml of an aqueous 37% formaldehyde solution are added. The reaction mass is kept for 2 hours, then cooled to room temperature, 25 ml of concentrated hydrochloric acid are added and stirred for 30 minutes. The mixture was extracted with methylene chloride, the organic layer was washed with water until neutral, the solvent was evaporated. After crystallization, 9.4 g of 11β, 17α, 21-triacetoxy-6-methylene-pregn-4-ene-3,20-dione are obtained in 92% yield. According to example 4 of this method, under similar conditions, from 1 g of 21-acetoxy-11β, 17α-bis (trifluoroacetyloxy) -pregn-4-ene-3,20-dione, 0.93 g of the 6-methylene derivative are obtained in 91% yield.

The main disadvantage of this method is the need for mandatory protection of all hydroxyl groups present in the structure of the hydrocortisone molecule with subsequent removal of the protection. So, when applying the method of exhaustive acetylation to protect, despite the high stepwise yields upon receipt of 11β, 17α, 21-triacetoxy-6α-methylpregn-4-ene-3,20-dione, effective removal of the acetyl group with the regeneration of the 11β-hydroxyl group is possible only using the biotechnological method, namely the microbiological transformation of bacterial cells by Arthrobacter mediolanus VKM-1388, which is carried out in 2 stages (enzymatic deprotection of the 11β-hydroxyl group and hydrolysis of the 17α-acetoxy group by the action of a 10% hydroxide solution sodium) with a total duration of the process of deacetylation of 72 hours. This significantly complicates the method.

Disclosure of invention

The technical problem in the claimed invention is the development of a simpler method for producing 6α-methylhydrocortisone or its esters from 21-hydrocortisone acetate while ensuring the yield of the target products of not less than 80%.

The simplification of the method is ensured by 1) through sequential functionalization reactions of the C ⁶ position of the molecule of 11-trifluoroacetate 21-hydrocortisone (III) acetate with the formation of 21-acetoxy-11-trifluoroacetyloxy-6-methylene-hydrocortisone (VI) at ambient temperature (i.e. E. at room temperature) without the use of heating the reaction masses; 2) by carrying out a continuous technological process of functionalizing the C ⁶ position of the molecule of 11-trifluoroacetate 21-hydrocortisone (III) acetate without isolating the intermediates IV and V of the target product (VI) at key stages of the synthesis and thereby eliminating not only the loss of these intermediates IV and V in mother liquors, but also mechanical losses, which usually take place in the technological operations of extraction, filtration, drying, etc .; 3) by achieving almost complete conversion of the starting products and high selectivity of the reactions of the formation of intermediates IV and V and the target product VI during the functionalization of the C ⁶ position of the molecule of 11-trifluoroacetate 21-hydrocortisone (III) acetate and thereby reducing the likelihood of adverse reactions; 4) due to the almost complete exclusion of the formation of by-products and the grinding of the reaction mass at the stages of removal of the protective groups. The result is a high yield of final products I, VIII and IX (from 80 to 94), counting from 21-hydrocortisone (II) acetate.

The technical problem is solved by the method of obtaining 6α-methylhydrocortisone or its esters of General formula (I)

where R = COCF ₃ or H, R ₁ = COCH ₃ or H

from 21-hydrocortisone acetate of the formula (II),

including preliminary protection of the 11β-hydroxyl group of the hydrocortisone 21-acetate molecule by esterification with trifluoroacetic acid anhydride under basic catalysis using catalytic amounts of catalyst, followed by C ⁶ -methyleneation of the resulting 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-pregn-4-ene -3,20-dione, catalytic isomerization of the 6-exomethylene bond of the obtained 6-methylene derivative into endomethylene with the formation of 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6-methylpregna-4,6-diene-3,20- dione, hydrogenation of the 6,7-double bond of the latter with the formation of 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6α-methylpregn-4-ene-3,20-dione, simultaneous, phased or sequential deprotection of 11β- and 21- hydroxyl groups by chemical solvolysis methods.

Below is the proposed scheme for the synthesis of 6α-methylhydrocortisone (I) or its esters (VIII and IX) from 21-hydrocortisone acetate (II).

The scheme includes 7 (for VIII) or 8 (for I and IX) chemical reactions that can be carried out in 3 or 4 technological stages, respectively. The chemical synthesis scheme includes: 1) protection of the 11β-hydroxyl group of the hydrocortisone (II) 21-acetate molecule by the formation of the 11β-trifluoroacetyloxy derivative (III); 2) enolization of the α, β-unsaturated ketone of ring A of compound III with the formation of 3,5-dienol ether (IV); 3) a three-component condensation reaction according to the Mannich type reaction with the formation of a mixture of C ⁶ -aminomethyl derivatives (V); 4) cleavage of the aminomethyl derivatives (V) via a CN bond to form a C ⁶ -methylene derivative (VI); 5) the reaction of catalytic isomerization of a 6-exomethylene bond into a 6,7-endomethylene bond with the formation of compound VII; 6) the reaction of catalytic hydrogenation of a 6,7-double bond with the formation of a mixture of 6α- and 6β-methyl diastereomers (VIII and VIIIa, respectively); 7) catalytic isomerization of the 6β-methyl group of compound VIIIa to 6α-methyl to form compound VIII; 8) the removal of the protective groups of the molecule of compound VIII completely with the formation of 6α-methylhydrocortisone (I) or partially with the hydroxyl group C ²¹ with the formation of compound IX.

For the esterification of the 11β-hydroxyl group of compound II, trifluoroacetic acid anhydride can be used as the trifluoroacetic acid derivative.

The esterification of the 11β-hydroxyl group of compound II is carried out in an aprotic solvent, using dialkyl ketones or cyclic ethers or dimethyl sulfoxide under basic catalysis using a catalyst in the amount necessary for complete conversion of the starting compound. In this case, the dialkyl ketone is selected from the group acetone, methyl ethyl ketone; the glycol ethers are selected from the group tetrahydrofuran, dioxane.

Dimethylaminopyridine is used as an esterification catalyst.

The enolization of the Δ ⁴ -3-ketosystem of compound III for esterification with the formation of the 3-alkoxy ester Δ ^3,5 -3-hydroxy derivative of the general formula IV is carried out by the action of protic acids.

Sulfonic acids are used as organic acids for the enolization of the Δ ⁴ -3-ketosystem of compound III.

Sulfonic acids are selected from the group comprising sulfosalicylic acid, p-toluenesulfonic acid.

The trialkyl orthoformate for the formation of the 3-alkoxy ester of Δ ^3,5 -3-hydroxy derivative of the general formula IV is taken in an amount sufficient to carry out the reaction to give formulas IV and V.

The formation of the 3-alkoxy ester Δ ^3,5 -3-hydroxy derivative of the general formula IV is carried out in a proton or aprotic solvent, which is used as aliphatic alcohols, cyclic ethers, aromatic hydrocarbons of a series of benzene or mixtures thereof.

As aliphatic alcohols, methanol or ethanol is used.

As cyclic ethers, dioxane or tetrahydrofuran is used.

As aromatic hydrocarbons, benzene or toluene is used.

As the trialkyl orthoformate, trimethyl orthoformate or triethyl orthoformate is used.

The 6-aminomethylation of the compounds of general formula IV is carried out in a protic or aprotic solvent, which is used aliphatic alcohols (e.g. methanol, ethanol), cyclic ethers (dioxane, tetrahydrofuran), aromatic hydrocarbons of the benzene series (benzene, toluene) or mixtures thereof.

The 6-aminomethylation of a compound of general formula IV is carried out in a trialkyl orthoformate medium or in a mixture of trialkyl orthoformate and an organic solvent without isolating a compound of general formula IV from the reaction mixture.

The deamination of a mixture of compounds of general formula V is carried out in an aprotic solvent or a mixture of aprotic solvents by the action of dilute sulfuric acid using an effective amount thereof.

Alkali or alkaline earth metal or ammonium salts are used as salts of hydrohalic acid, and hydrochloric or hydrobromic acids are used as hydrohalic acid.

The isomerization of the 6-exomethylene bond of the compound of formula VI to form the 6,7-endomethylene bond of compound VII is carried out using palladium or palladium oxide adsorbed on carbon as a catalyst.

Hydrogenation of the 6.7-endomethylene bond of compound VII is carried out using palladium or palladium oxide adsorbed on carbon as a catalyst.

The removal of the protective group of the 21-hydroxyl group is carried out by the method of acid solvolysis, both with or without isolation of the hydrogenation product of the formula VIII from the reaction mixture to form 11β-trifluoroacetate 6α-methylhydrocortisone IX (R = COCF ₃ , R ₁ = H).

The deamination of a mixture of compounds of General formula V is carried out without isolation from the reaction mass.

The essence of the claimed invention relating to the production of 6α-methylhydrocortisone or its esters is that in order to increase the yield and simplify the process, the 21-hydrocortisone acetate is first subjected to preliminary protection of the 11β-hydroxyl group, then to methyleneation of the C ⁶ position, isomerization of the exomethylene group at C6 in 6,7-endomethylene, restoration of the 6,7-methylene bond of the 6-methyl-Δ ⁶ group with the formation of the 6α-methyl derivative, the removal of the protective groups.

The advantages of the proposed method are as follows:

- trifluoroacetate protection of the 11β-hydroxyl group of hydrocortisone 21-acetate and selective deesterification of the by-product of the esterification reaction of 11,17-bis (trifluoroacetate) 21-hydrocortisone acetate with 17α-hydroxyl group regeneration is carried out in one chemical stage without isolation of the target product;

- conducting condensation with the Mannich reagent in the presence of free hydroxyl at C ¹⁷ under mild conditions (at room temperature) eliminates the appearance of undesirable products (dehydration of the 17α-hydroxy group with the formation of the Δ ¹⁶ bond and D-homo-channeling with the formation of the 6-membered ring D), which confirmed by NMR spectroscopy;

- the regioselectivity of the cleavage of the Mannich steroid base with mineral acid exclusively via the C-N bond is ensured by the presence in the reaction medium of a salt of hydrohalic acid and an alkali, or alkaline earth metal, or ammonium;

- the absence of the need to use multiple crystallization of 6α-methylhydrocortisone or chromatography to purify it, since the removal of the protective groups leads to a slight appearance of side compounds; the content of the main substance in the product is 90-95% (HPLC), which meets the requirements of, for example, the subsequent process of introducing a 1,2-double bond (chemical or biotechnological method);

- reactions of trifluoroacetylation, enolesterification, condensation with Mannich’s reagent, deamination of Mannich’s steroid base are carried out at room temperature;

- a significant reduction in the losses of the main product at the stages, which is ensured by the high selectivity of chemical reactions and the possibility of combining the chemical stages into one technological process without isolation of intermediates, can significantly increase the achieved total yield of 6α-methylhydrocortisone and its esters from hydrocortisone 21-acetate to 80.1% and 82.9-93.6%, respectively.

The implementation of the invention

The method of obtaining 6-methylhydrocortisone or its esters of the general formula (I) is carried out according to the following scheme (Figure 1).

21-hydrocortisone acetate (21-acetoxy-11β, 17α-dihydroxy-pregn-4-en-3,20-dione) of the formula (II),

protect the 11β-hydroxyl group by esterification with trifluoroacetic anhydride in an aprotic solvent under basic catalysis in the presence of a catalytic amount of dimethylaminopyridine. For this, 1-1.5 mol of trifluoroacetic anhydride and not more than 0.1 mol of dimethylaminopyridine are used per 1 mol of hydrocortisone 21-acetate (21-acetoxy-11β, 17α-dihydroxy-pregn-4-en-3,20-dione). Obtained with a yield of up to 99.95%, 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-pregn-4-ene-3,20-dione (III) is subjected to enolesterification of the Δ ⁴ -3-keto ring system by the action of alkyl orthoformate in the presence of sulfoaromatic acid (sulfosalicylic or p-TSC), in the amount necessary for the enolization of Δ ⁴ -3-keto group. Then, the obtained 3,5-dienol ether (IV) is subjected to catalytic condensation with a Mannich reagent formed from formaldehyde and N-methylaniline in situ, with the formation of the 6- (N, N-disubstituted) -aminomethyl derivative (V), followed by its deamination and conversion of the obtained 6-methylene derivative (VI) in the 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6α-methylpregn-4-ene-3,20-dione (VIII) via the formation of the 6-methyl-6,7-dehydro-isomer (VII), moreover, condensation with the Mannich reagent can be carried out both with preliminary isolation and without isolation of 3,5-dienole Fira (IV), as well as with or without isolation of the condensation product (V) from the reaction mixture, and the reduction of the 6-methylene group of compound VI to the 6α-methyl group of compound VIII - with or without isolation 6 methyl 6,7-dehydro-intermediate VII. After this, by chemical solvolysis of 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6α-methylpregn-4-ene-3,20-dione (VIII), 6α-methylhydrocortisone (I) or its ester (IX) is obtained. Moreover, the removal of the protective groups of compound VIII with the formation of compound I can be realized simultaneously under the conditions of basic catalysis or stepwise through the formation of compound IX by solvolysis under acid catalysis and subsequent solvolysis of the 11-trifluoroacetoxy group of compound IX under the conditions of basic catalysis.

The total achieved yield of 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6α-methylpregn-4-ene-3,20-dione (VIII) from hydrocortisone (II) 21-acetate is 93.6%.

The total achievable yield of 6-methylhydrocortisone (IX) 11-trifluoroacetate from hydrocortisone (II) 21-acetate is 82.9%.

The total achieved yield of 6α-methylhydrocortisone (I) from 21-hydrocortisone (II) acetate is 80.1%.

The protection of the 11β-hydroxyl group of the hydrocortisone (II) 21-acetate molecule is carried out by the formation of trifluoroacetic acid ester in an organic solvent under basic catalysis. This protection option is most optimal, since the esterification reaction proceeds under mild conditions with an almost quantitative yield. In addition, the trifluoroacetate protecting group, stable in an acidic environment, is easily removed if necessary by hydrolysis under the conditions of basic catalysis. Acylation is carried out using the minimum required amount of trifluoroacetic anhydride under anhydrous conditions in an organic aprotic solvent. One skilled in the art of organic chemistry, as well as from the prior art, knows that instead of trifluoroacetic acid anhydride, trifluoroacetic acid chloride can be used in this process, and as the aprotic solvent in carrying out the invention, a method for producing 6α-methyl hydrocortisone or its esters, can be used any aprotic (non-polar, low-polar and polar) solvents suitable for the reaction and inert with respect to the acylating reagent and catalyst, such as dialkylket us (e.g., acetone, methyl ethyl ketone), cyclic ethers (e.g., dioxane, tetrahydrofuran), and others known in the art, to ensure completeness of reaction. As the main catalyst, heteroaryl compounds (e.g., pyridine or its derivatives), or alkylamines (e.g., triethylamine), or other organic bases, or mixtures thereof, commonly used to catalyze acylation reactions of hydroxyl groups known in the art, can be used.

The trifluoroacetylation reaction of compound II proceeds regionally, preferably esterifying the secondary hydroxyl group at the C ¹¹ atom of compound II. To ensure the progress of the reaction, the necessary minimum amount of trifluoroacetic anhydride is 1 mol per 1 mol of steroid. For a more complete and quick reaction, a small excess of the acylating agent is used (~ 1.2-1.5 mol of TFA per 1 mol of steroid). However, this condition is not required. The use of an excess amount of the reagent leads to partial trifluoroacetylation of the tertiary hydroxyl group at C ¹⁷ of reaction product III with the formation of 10-20% impurities of 11β, 17α-dithrifluoroacetyloxy derivative (IIIa).

However, under the conditions of our method, the C ¹⁷ trifluoroacetic acid ester is removed easily and selectively with the regeneration of the 17α-hydroxyl group and preservation of the trifluoroacetate protection of the 11β-hydroxyl group, which is achieved by treating the reaction mass with triethylamine in the minimum required amount. The yield of 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy pregn-4-en-3,20-dione (III) in the step is from 96 to 99.95%.

Synthesis of 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6-methylene-pregn-4-ene-3,20-dione (VI) from 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-pregn-4-en-3, 20-dione (III) includes three sequential chemical processes that can be carried out with the isolation of any of the intermediates of the general formulas IV and V or without isolation according to the following variants of the chemical synthesis scheme:

Variant of scheme 1: III-IV-V-VI; or

Variant of scheme 2: III-IV- [V] -VI; or

Variant of scheme 3: III- [IV] -V-VI; or

Variant of scheme 4: III- [IV] - [V] -VI.

The most optimal and preferred option is scheme 4.

To increase the nucleophilicity atom C ^6, and thereby reduce the likelihood of side condensation processes on other reaction centers (e.g., at C ²⁾ in the first synthesis step is conducted enolization reaction of Δ ⁴ -3-ketosistemy ring A of the compound III to form 3,5- dienol ether of General formula IV. The enolesterification reaction of the Δ ⁴ -3-ketosystem of ring A, both with the release of enol ether IV (options 1 and 2), and without isolation of enol ether IV (options 3 and 4) from the reaction mixture, can be carried out with or without the use of a solvent. The enolesterification reaction of the Δ ⁴ -3-ketosystem with a solvent is carried out by the action of formic acid trialkyl orthoester (e.g., trimethyl orthoformate or triethyl orthoformate) using the minimum amount of tri-alkyl orthoformate necessary to ensure the reaction (at least 1 mol per 1 mol of steroid). For a more complete and quick reaction (15-30 minutes, but not more than 2 hours), an excess of alkyl orthoformate is used (from 2.5 to 9 mol per 1 mol of steroid). However, this condition is not required. The reaction is carried out under anhydrous conditions in an appropriate protic solvent — an aliphatic alcohol (in methanol or ethanol) or in an aprotic solvent selected from the group of cyclic ethers (for example, dioxane or tetrahydrofuran) or aromatic hydrocarbons of the benzene group (for example, benzene or toluene). The enolesterification reaction of the Δ ⁴ -3-ketosystem without the use of a solvent is carried out in a trialkyl orthoformate reagent medium.

As an enolization catalyst, any protic acid can be used to provide the enolization of the Δ ⁴ -3-keto ring system of ring A of compound III to form 3,5-dienol, as known to those skilled in the art. It is preferable to use organic acids, for example sulfoaromatic (sulfosalicylic acid, p-TSC or the like), both anhydrous and in the form of crystalline hydrate. When carrying out the reaction without isolating enol ether IV (option 3 and 4) from the reaction mixture, it is preferable to use p-TSC, which is also a catalyst for the subsequent aminomethylation reaction.

In the synthesis of compounds of general formula V, the introduction of the substituent at position C ^{6 is} carried out by the method of three-component condensation according to the Mannich reaction using a Mannich reagent formed in situ by the interaction of formaldehyde and N-methylaniline or other secondary amines, suitable for the formation of 6- (N, N-disubstituted) -aminomethyl compounds and known in the art, for example, N-ethylaniline, diphenylamine, N-methyl-p-toluidine and the like. [GB 1280570, 1972].

According to the present invention, the Mannich reaction with the formation of compounds of general formula V can be carried out both with the isolation of product IV from the reaction mass (options 1 and 3), and without isolation (options 2 and 4). However, it is preferable to carry out the Mannich reaction without isolating enol ether IV (options 3 and 4) in a trialkyl orthoformate medium without using solvents both at the enol ether preparation stage and at the aminomethylation stage. Both processes are carried out at room temperature. At the end of the reaction to obtain enolether IV, N-methylaniline and a 30-40% aqueous formaldehyde solution are added to the reaction mass.

The aminomethylation reaction is non-stereoselective: a mixture of 6α- and 6β-diastereomeric Mannich steroid bases (Va and Vb, respectively) is formed. The aminomethylation reaction can be carried out in a medium containing a cyclic ether (for example, dioxane or tetrahydrofuran) or an aromatic hydrocarbon of the benzene group (for example, benzene or toluene), and these solvents or mixtures thereof can be added to the reaction mass containing compounds of the general formula V after completion of the aminomethylation reaction. A solution of a compound of general formula V in an aromatic hydrocarbon is washed with 1-5% aqueous ammonia to remove p-TSC and then with water. This solution in an aromatic hydrocarbon can be used in the next deamination step without extracting a mixture of isomers of the general formula V (options 2 and 4) or the specified mixture of Mannich’s isomeric steroid bases can be extracted from the solution (options 1 and 3). However, the latter is impractical, since both isomers can be subjected to a subsequent deamination reaction with the formation of 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6-methylene-pregn-4-ene-3,20-dione (VI) without isolation and separation.

The region-directed cleavage of the Mannich steroid base of general formula V via CN is carried out by the action of concentrated or diluted mineral acids, for example, 30-35% hydrochloric acid or 50-70% sulfuric acid (preferably 55-60% sulfuric acid) in an aromatic hydrocarbon series of benzene (toluene or benzene) with the addition of an aprotic solvent miscible with water, for example, dialkyl ketone (acetone or methyl ethyl ketone) or cyclic ether (dioxane or tetrahydrofuran). The addition of a proton solvent, in particular lower aliphatic alcohol (methyl or ethyl alcohol, or others) can lead, firstly, to a partial cleavage of the Mannich steroid base of the general formula V via CC bond with the regeneration of the starting compound III, and secondly , to the removal (removal) of the ester group at the C21 atom in both the resulting by-product III and the target product VI. The regionally directed cleavage reaction of the Mannich steroid base of general formula V via the C-N bond can be carried out in the presence of mineral salts, for example, alkali metal or alkaline earth metal halides, or ammonium halides.

To carry out the deamination reaction without isolating the mixture of Va and Vb isomers (options 2 and 4), the solution of the Mannich (V) steroid base mixture in aromatic hydrocarbon is diluted with the minimum amount of dialkyl ketone (at least 40% vol.), 55-60% sulfuric acid solution is added and mineral salt. The reaction is carried out at room temperature. At the end of the reaction, the reaction solution is washed with acid. The solvent was evaporated in vacuo or at atmospheric pressure. The remainder of the aromatic hydrocarbon can be removed by azeotropic distillation with any convenient solvent, forming an azeotropic mixture with an aromatic hydrocarbon and miscible with water, for example, ethyl or methyl alcohol. The reaction product is isolated by dissolving the residue after distillation in alcohol (ethyl or methyl) and diluting the resulting solution with water.

The achieved yield of 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6-methylenepregn-4-en-3,20-dione (VI) from hydrocortisone (II) 21-acetate is:

according to option 1 (II-III-IV-V-VI) - from 87.7 to 91.8%;

according to option 2 (II-III-IV- [V] -VI) - from 88.3 to 92.5%;

according to option 3 (II-III- [IV] -V-VI) - up to 93.8%;

according to option 4 (II-III- [IV] - [V] -VI) - up to 97.8%.

It should be noted that the deamination reaction in the medium of aprotic organic solvents or their mixtures, as well as protonic solvents, is non-selective using the action of a mineral acid (sulfuric or hydrochloric) without the addition of an inorganic salt: deamination is accompanied by deamination with the regeneration of compound III. The use of inorganic salts in combination with mineral acid is not obvious, it is not known from the prior art, to ensure the regional directionality of the splitting of steroid Mannich bases on the C-N bond has not been previously used. The reaction without the addition of an inorganic salt leads to a decrease in the yield of the product. Carrying out the reaction by the action of concentrated hydrochloric acid in tetrahydrofuran and ethanol (a literary version of the splitting of Mannich base [RU 2297423, 2007]), although it leads to a reduction in the duration of the process, this yields product VI, which is not suitable for the subsequent hydrogenation reaction to form compound VIII without further purification, for example, crystallization. As a result of purification by crystallization, the yield at the stage of synthesis of compound VI is only 60-65%.

The process of obtaining 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6α-methyl-pregn-4-ene-3,20-dione (VIII) from 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6-methylenepregn-4 en-3,20-dione (VI) can be carried out in three stages without isolation of intermediates VII and VIIIa or with isolation of compound VII. Polar solvents such as lower carboxylic acids (e.g., acetic acid) and aliphatic alcohols (e.g., methanol, ethanol) can be used as the medium for the three-stage process.

The first step is the catalytic isomerization of the exomethylene double bond of compound VI into the endomethylene 6,7-double bond of compound VII.

The second stage is the catalytic hydrogenation of the endomethylene bond of the 6-methyl-Δ ⁶ system of compound VII by hydrogen transfer in the presence of cyclohexene. Isomerization (first step) and hydrogenation (second step) in the claimed invention is carried out in the presence of traditionally used catalysts, for example palladium adsorbed on carbon (palladium catalyst) and others. The reaction is carried out at the boiling point of the reaction mixture. The process is controlled by determining the composition of the reaction mass by absorption in the UV spectrum at wavelengths corresponding to the absorption maxima of compound VIII (at 239 nm) and compound VII (at 286 nm). At the end of the recovery process, the catalyst is filtered off from the hot reaction mass. The hydrogenation reaction proceeds non-stereoselectively: a mixture of 6α- and 6β-diastereomeric methyl derivatives VIII and VIIIa is formed, respectively.

The third stage, the isomerization of the side 6β-methyl isomer VIIIa to the target product VIII under conditions of acid catalysis, is carried out without isolating the mixture of reduction products of 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6α-methylpregn-4-en-3,20- dione (VIII) and 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6β-methylpregn-4-en-3,20-dione (VIIIa) from the filtrate and their separation. Concentrated hydrochloric acid is used as an isomerization catalyst. The reaction is carried out at room temperature for 30 minutes. The reaction product is isolated by diluting the residue after distilling off the solvent with water.

The yield of 21-acetoxy-11-trifluoroacetyloxy-6α-methylhydrocortisone (VIII) in the step is 95-96%.

The removal of ester groups at C ¹¹ and C ²¹ can be carried out simultaneously or in stages by hydrolysis reactions.

The complete removal of ester groups at C ¹¹ and C ²¹ with the formation of compound I is carried out by hydrolysis catalyzed by a strong base. The reaction is carried out in methanol. As a strong base, a catalytic amount of alkali metal hydroxide (sodium or potassium) is used in the form of a 10% aqueous solution.

Get 6α-methylhydrocortisone (I) with a basic substance content of 90-95%. The product can be used in the chemical or biotechnological process of 1,2-dehydrogenation, as well as in the synthesis of 21-esters, without further purification.

The removal of the ester group at C ²¹ of compound VIII while maintaining the 11β-trifluoroacetoxy group and the formation of 6α-methylhydrocortisone (IX) 11-trifluoroacetate is carried out under conditions of acid catalysis.

It should be noted that the removal of the trifluoroacetyl group at C ¹¹ of compound VIII while maintaining the 21-acetoxy group and the formation of 6α-methylhydrocortisone 21-acetate can be carried out by the action of triethylamine in methanol under the conditions of the patent [US 4330541, 1982, example 1].

The claimed invention is illustrated by the following examples. The duration of the reactions, the yield of products, the use of inorganic salts are given for illustration only, are shown to illustrate the achievement of the specified technical result. Instead of these salts, any water-soluble salts formed by hydrohalic acids, for example, chlorides and bromides of lithium, sodium, potassium, magnesium, barium, ammonium and others, can be used without loss of effectiveness.

The catalyst is taken in an amount that ensures the reaction taking into account various technological factors of the reaction, especially the dependence of the temperature / duration of the reaction (the more the catalyst, the faster the reaction, the higher the temperature - also faster, the catalyst can be smaller, but various by-products can form )

21-hydrocortisone (II) acetate, CAS No. 50-03-3, C ₂₃ H ₃₂ O ₆ (USP quality, or EP, or BP), is commercially available, can be purchased, for example, from Steraloids (USA), or from other manufacturers. 4-Dimethylaminopyridine (≥99.0%), trifluoroacetic anhydride (≥99.0%), trimethylorthoformate (≥99.0%), triethylorthoformate (≥98.0%), 5-sulfosalicylic acid (≥99.0%), p-toluenesulfonic acid (≥98.0%) , N-methylaniline (≥98.0%), cyclohexene (≥99.0%) are commercially available, were purchased from Sigma-Aldrich Co. Other reagents, solvents and inert gases are commercially available, were purchased from Russian manufacturers.

Distilled water was used to prepare aqueous solutions of acids, sodium chloride, ammonia, and sodium hydroxide. To isolate products at the end of reactions by pouring the reaction masses into water, as well as to wash precipitates and extracts in organic solvents, we used drinking tap water, unless otherwise specified.

All procedures, unless otherwise specified, were carried out at room temperature or ambient temperature, that is, in the range from 20 to 25 ° C. For processes requiring lower temperatures than room temperature, cooling was provided with cold tap water (in the range of 10 to 20 ° C), or a mixture of crushed ice and cold water (in the range of 5 to 10 ° C) or a mixture of crushed ice and chloride calcium (at a temperature below 5 ° C). An electric heating mantle was used for heating during reactions at temperatures above room temperature and for evaporating the solvents at atmospheric pressure.

Labortechnik AG), при остаточном давлении 0,35±0,05 кгс/см² (35±5 кПа) и температуре воды в бане в диапазоне от 35-50°С в зависимости от природы упариваемого растворителя.Evaporation of solvents in vacuo was carried out using a Rotavapor (

Labortechnik AG), with a residual pressure of 0.35 ± 0.05 kgf / cm ² (35 ± 5 kPa) and a water temperature in the bath in the range from 35-50 ° C, depending on the nature of the evaporated solvent.

Drying of the product crystals to constant weight was carried out at a temperature of 35-45 ° C at atmospheric pressure or using a vacuum oven at a residual pressure of 0.35 ± 0.05 kgf / cm ² (35 ± 5 kPa).

A universal indicator paper with a range of values from 0 to 12 (Lahema, Czech Republic) was used to determine the pH of the washings.

Dry (anhydrous) solvents (acetone, triethylamine, methanol, ethanol) were obtained by well-known methods of organic chemistry.

Column chromatography was performed on a column (16 × 650 mm) using silica gel of the brand Silica gel 60 (0.040-0.063 mm) (Merck, Germany).

The reaction progress was monitored by thin layer chromatography (TLC) using Silica gel 60 F254 plates (Merck, Germany). UV spectroscopic determination of the end of the isomerization reactions of the 6-methylene bond of compound VI and the hydrogenation of the 6.7-double bond of compound VII was carried out on a Perkin Elmer Lambda 40 UV spectrophotometer, measuring the ratio of the optical densities of the reaction solution sample at wavelengths D ₂₈₆ , D ₂₅₈ and D ₂₃₉

The structure and purity of all isolated compounds was confirmed by at least one of the following methods: TLC (plates for TLC Silica gel 60 F254 (Merck, Germany)), mass spectrometry, elemental analysis or nuclear magnetic resonance (NMR), high performance liquid chromatography (HPLC).

Labortechnik AG).The melting point of the isolated compounds was determined on a device to determine the melting point of M-565

Labortechnik AG).

¹ H and ¹³ C NMR spectra were determined on a Bruker Avance-400 spectrometer (Bruker BioSpin GmbH) with an operating frequency of 400 MHz and 100.6 MHz, respectively, using deuterated chloroform (99.8% D, Sigma-Aldrich), or deuterated dimethyl sulfoxide (99.9% D, Sigma-Aldrich) as a solvent relative to tetramethylsilane (TMS NMR grade ≥99.9%, Sigma-Aldrich) as an internal standard, in parts per million (ppm).

Elemental analysis was performed using a Vario MICRO Cube analyzer (Elementar Analysensysteme, GmbH).

HPLC analysis was performed on a Knauer Smartline (Germany) chromatograph equipped with a gradient HPLC pump, a cell for a thermostated column, an injector, a Smartline 2600 diode array detector, an integrator-computer, at a temperature of 24 ° C, a flow rate of 1 ml / min and UV detection at a wavelength of 254 nm. Kromasil® 100-5С18 was used as a stationary phase, and a solution of Н ₂ О in acetonitrile was used as a mobile phase - 35% (by volume).

Example 1. Obtaining 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy pregn-4-en-3,20-dione (III)

Option 1

To a suspension of 10 g (24.722 mmol) of 21 hydrocortisone (II) acetate in 80 ml of dry acetone was added 0.3 g (2.455 mmol) of dimethylaminopyridine. Then, at a temperature not exceeding 10 ° C., 4.2 ml (6.28 g, 29.9 mmol) of trifluoroacetic anhydride were added. The reaction mass was warmed to room temperature (20-25 ° C) and stirred for 1.5 hours. At the end of the reaction, 10 ml (7.28 g, 71.94 mmol) of dry triethylamine were added, kept for 15 minutes. Then 4.7 ml (4.93 g, 82.1 mmol) of acetic acid were added dropwise to a pH of 6 and the reaction mass was slowly poured into 240 ml of water at a temperature of 8-10 ° C. The suspension was kept under vigorous stirring for 1 h at the same temperature. The precipitate was filtered off, washed with water to pH ~ 7 wash water, dried to constant weight. Received 11.88 g (23.736 mmol) of compound III with a yield of 96.01%. Mp. 204-206 ° C (lit. 206-207 ° C [DE 1125422, 1962]). ¹ H NMR (400 MHz, CDCl ₃ , δ, ppm): 5.70 (1H, s, H-4), 5.67 (1H, m, H-11), 5.09 (1H, d, J = 17.4 Hz , H-21), 4.67 (1H, d, J = 17.4 Hz, H-21), 2.14 (3H, s, CH ₃ C (O)), 1.26 (3H, s, H-19), 0.80 (3H , s, H-18); ¹³ C NMR (100.6 MHz, CDCl ₃ , δ, ppm): 204.9, 198.8, 170.5, 170.1, 122.9, 89.1, 75.4, 67.6, 54.9, 51.4, 47.0, 38.5, 35.5, 35.4, 34.5, 33.5, 32.2, 31.8, 31.7, 23.5, 20.6, 20.5, 16.4 .. Found: C 60.47%; H, 6.24%; C ₂₅ H ₃₁ F ₃ O ₇ . Calculated: C 59.99%; H 6.24%.

Option 2

The reaction mixture, consisting of 100 g (247.22 mmol) of hydrocortisone 21-acetate, 500 ml of tetrahydrofuran and 10 g (81.833 mmol) of dimethylaminopyridine, was cooled with stirring to a temperature of 3-5 ° C. Then, 42 ml (62.79 g, 298.96 mmol) of trifluoroacetic anhydride were slowly added to the reaction mixture in a stream of dry nitrogen so that the temperature of the reaction mixture did not exceed 7-8 ° C. After that, the bath with the cooling mixture was removed. The reaction mass was stirred until the temperature in the mass rose to room temperature (self-heating). At the end of the exposure, the reaction mass was heated to boiling point and tetrahydrofuran was distilled off at atmospheric pressure until the temperature in vapors reached 70 ° С.

The bath for heating was changed to a bath for cooling. The bottoms suspension was cooled to a temperature of 10 ° C and 55 ml (40.04 g, 395.69 mmol) of triethylamine were added via a dropping funnel to a pH of 8-9. After that, the cooling bath was removed, the suspension was stirred at room temperature for 1 h.

Upon completion of the reaction, the reaction mass was poured with vigorous stirring into 600 ml of water, cooled to a temperature of 0-2 ° C. It was stirred at the same temperature for 10-15 minutes and 11 ml (11.54 g, 192.17 mmol) of acetic acid were added to neutralize triethylamine. The suspension was stirred at a temperature of 20-25 ° C for 20 minutes. The precipitate was filtered and washed on the filter with water to a pH of ~ 7 wash water, dried to constant weight.

123.67 g (247.09 mmol) of compound III were obtained in a yield of 99.95%.

Option 3

The esterification reaction was carried out similarly to option 1, using dimethyl sulfoxide as the aprotic solvent. From 1 g (2.472 mmol) of 21 hydrocortisone (II) acetate, 1.12 g (2.238 mmol) of compound III were obtained in a yield of 90.53%.

A. Scheme, option 1: III-IV-V-VI

Example 2. Obtaining 3-methoxy-21-acetoxy-11β-trifluoroacetyloxy-17α-hydroxy-pregna-3,5-dien-20-one (IVa, R = CH ₃ )

Option 1

To a suspension of 5 g (9.99 mmol) of 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-pregn-4-ene-3,20-dione (III) in a mixture of 20 ml of dry methanol and 10 ml (9.676 g, 91 , 18 mmol) of trimethylorthoformate was added 0.125 g (0.726 mmol) of anhydrous p-TSC. The reaction mass was stirred for 30 minutes at room temperature. The excess p-TSC was neutralized with triethylamine, poured into 200 ml of ice water, the suspension was cooled to 0-2 ° C and kept under stirring for 2 hours. The precipitate was filtered off, washed on the filter with water to pH ~ 7 washing water and a minimum amount of methanol, dried to constant weight. Received 5.04 g (9.795 mmol) of compound IVa in the form of white crystals with a yield of 98.05%. Mp 177 ° C (decomp.). ¹ H NMR (400 MHz, CDCl ₃ , δ, ppm): 5.72 (1H, s, H-11), 5.15-5.09 (3H, m, H-4, H-6 and H-21), 4.68 (1H, d, J = 17.4 Hz, H-21), 3.57 (3H, s, CH ₃ O), 2.16 (3H, s, CH ₃ C (O)), 1.03 (3H, s, H-19 ), 0.80 (3H, s, H-18); ¹³ C NMR (100.6 MHz, CDCl ₃ , δ, ppm): 205.0, 170.6, 155.4, 141.1, 121.0, 116.7, 97.5, 89.3, 75.8, 67.6, 54.3, 52.6, 50.6, 47.0, 38.7, 35.5, 34.6, 33.6, 31.4, 28.4, 24.8, 23.5, 21.1, 20.5, 16.3.

Option 2

To a suspension of 1 g (1.998 mmol) of compound III in a mixture of 4.7 ml of dry methanol and 0.55 ml (0.532 g, 5.015 mmol) of trimethylorthoformate was added 0.011 g (0.05 mmol) of anhydrous sulfosalicylic acid (CCK). The reaction mass was stirred for 2 hours at room temperature. Excess CCK was neutralized with triethylamine. The reaction mass was diluted with ice water 3 times and kept with stirring for 30 min at a temperature of 8-10 ° C. The precipitate was filtered off, washed on the filter with water to a pH of ~ 7 wash water and a minimum amount of methanol, dried to constant weight. 0.96 g (1.816 mmol) of compound IVa were obtained in a yield of 90.89%.

Example 3. Obtaining 3-ethoxy-21-acetoxy-11β-trifluoroacetyloxy-17α-hydroxy-pregna-3,5-dien-20-one (IVb, R = CH ₂ CH ₃ )

Under the conditions of obtaining compound IVa (option 1), 20 ml of anhydrous ethanol and 10 ml (8.91 g, 60.121 mmol) of triethyl orthoformate were used, from 5 g (9.99 mmol) of 21-acetoxy-11β-trifluoroacetyloxy-17α-hydroxy-peptide 4-en-3,20-dione (III) gave 4.94 g (9.346 mmol) of compound IVb in 93.55% yield. Mp 160 ° C (decomp.). ¹ H NMR (400 MHz, CDCl ₃ , δ, ppm): 5.72 (1H, s, H-11), 5.17-5.07 (3H, m, H-4, H-6 and H-21), 4.68 (1H, d, J = 17.4 Hz, H-21), 3.76 (2H, q, J = 7.0 Hz, CH ₂ O), 2.16 (3H, s, CH ₃ C (O)), 1.28 (3H, t, J = 7.0 Hz, С Н ₃ СН ₂ О), 1.03 (3Н, s, Н-19), 0.80 (3Н, s, Н-18); ¹³ C NMR (100.6 MHz, CDCl ₃ , δ, ppm): 205.0, 170.6, 154.5, 141.3, 116.4, 98.0, 89.3, 75.8, 67.6, 62.2, 59.5, 52.6, 50.6, 47.0, 35.5, 34.8, 33.6, 31.4, 28.4, 25.0, 23.5, 21.1, 20.5, 16.3, 14.6.

Example 4. Obtaining 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6ξ- (N-methyl-N-phenylaminomethyl) pregn-4-en-20-one (V)

To a suspension of 10 mmol of 3,5-dienol ether (IVa or IVb) in 15 ml of absolute alcohol (methanol or ethanol, respectively), 2.15 ml (2.122 g, 19.8 mmol) of N-methylaniline was added. Then, with stirring, a solution of 0.125 g (0.725 mmol) of p-TSC in 1.25 ml of the corresponding alcohol and 2 ml (2.18 g, 0.807 g in 100%, 26.86 mmol) of a 37% aqueous solution was added dropwise. formaldehyde. The reaction mass was stirred at 20 ° C for 2 hours. At the end of the reaction, the reaction mass was diluted with 20 ml of the corresponding alcohol and poured slowly with vigorous stirring into 110 ml of water. The precipitate was filtered off, washed with water to pH ~ 7 wash water, dried to constant weight. Compound V (mixture of isomers) was obtained in quantitative yield. Without purification and separation of the epimers used in the next stage. To obtain an analytically pure sample method used to column chromatography on SiO ₂ pretreated with triethylamine.

Mp = 112-117 ° C.

6α-Isomer (Va), ¹ H NMR (400 MHz, CDCl ₃ , δ, ppm): 7.28-7.22 (2H, m, H-3,5 (NPh)), 6.79-6.69 (3H, m , H-2,4,6 (NPh)), 5.87 (1H, s, H-4), 5.70 (1H, m, H-11), 5.08 (1H, d, J = 17.4 Hz, H-21) , 4.66 (1H, d, J = 17.4 Hz, H-21), 3.70 (1H, dd, J = 3.1, 14.5 Hz, NCH ₂ ), 3.14 (1H, dd, J = 10.4, 14.5 Hz, NCH ₂ ) 2.97 (3H, s, NCH ₃ ), 2.16 (3H, s, CH ₃ C (O)), 1.19 (3H, s, H-19), 0.80 (3H, s, H-18).

6β-Isomer (Vb), ¹ H NMR (400 MHz, CDCl ₃ , δ, ppm): 7.28-7.22 (2H, m, H-3,5 (NPh)), 6.79-6.69 (3H, m , H-2,4,6 (NPh)), 5.83 (1H, s, H-4), 5.70 (1H, m, H-11), 5.07 (1H, d, J = 17.4 Hz, H-21) , 4.69 (1H, d, J = 17.4 Hz, H-21), 3.76 (1H, dd, J = 10.8, 14.5 Hz, NCH ₂ ), 3.32 (1H, dd, J = 4.1, 14.5 Hz, NCH ₂ ) 3.02 (3H, s, NCH ₃ ), 2.16 (3H, s, CH ₃ C (O)), 1.36 (3H, s, H-19), 0.83 (3H, s, H-18); ¹³ C NMR (100.6 MHz, CDCl ₃ , δ, ppm): 204.8, 198.7, 170.6, 170.2, 148.5, 129.4, 125.4, 116.8, 112.3, 89.1, 75.1, 67.6, 57.2, 54.7, 51.3, 46.8, 42.3, 39.7, 38.1, 37.8, 35.5, 34.5, 33.6, 33.3, 27.9, 23.3, 22.8, 20.5, 16.4. Found: C 64.08%; H 6.52%, N 2.15%, C ₃₃ H ₄₀ F ₃ NO ₇ . Calculated: C 63.96%; H, 6.51%; N, 2.26%.

Example 5. Obtaining 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6-methylene-pregn-4-en-3,20-dione (VI)

Option 1

To a suspension of 2.5 g (4.034 mmol) of a mixture of 6α and 6β isomers of compound V in a mixture of 30 ml of toluene and 15 ml of methyl ethyl ketone was added 1 g (11.515 mmol) of lithium bromide and 2 ml (3 g, 1.798 g in 100%, 18.332 mmol) 60% sulfuric acid.

The reaction mass was kept at room temperature for 1 h and transferred to a separatory funnel. The acidic layer was separated, extracted with toluene, and the extract was combined with the organic layer. The toluene solution was washed with 60% sulfuric acid, water to a pH of 6-7, 5% aqueous ammonia and water until neutral. Then the solution was clarified with activated carbon and evaporated, removing the remaining toluene with methanol. The bottom residue was poured into 70 ml of water, the suspension was kept at a temperature of 8-10 ° C, the precipitate was filtered. 1.94 g (3.785 mmol) of compound VI were obtained with a yield of 93.83%. Mp 191-193 ° C. ¹ H NMR (400 MHz, CDCl ₃ , δ, ppm): 5.86 (1H, s, H-4), 5.68 (1H, m, H-11), 5.09-5.01 (3H, m, CH ₂ = and H-21), 4.69 d (1H, d, J = 17.4 Hz, H-21), 2.15 s (3H, s, CH ₃ C (O)), 1.18 (3H, s, H-19), 0.80 (3H, s, H-18); ¹³ C NMR (100.6 MHz, CDCl ₃ , δ, ppm): 204.9, 199.2, 170.6, 168.2, 144.1, 120.6, 115.4, 89.0, 75.4, 67.6, 53.7, 51.7, 46.9, 39.7, 38.7, 35.4, 34.7, 34.5, 33.3, 31.9, 23.4, 20.5, 19.9, 16.3. Found: C 61.31%; H, 6.35%; C ₂₆ H ₃₁ F ₃ O ₇ . Calculated: C 60.93%; H, 6.10%.

Option 2

To a suspension of 0.5 g (0.807 mmol) of a mixture of 6α and 6β isomers of compound V in a mixture of 10 ml of toluene and 10 ml of methanol was added 0.32 ml (0.376 g) of 35% hydrochloric acid (0.13 g of hydrogen chloride, 3.556 mmol) hydrochloric acid and 1 ml of water. The reaction mass was stirred for 1 h at room temperature and transferred to a separatory funnel. The acidic layer was separated, extracted with toluene, and the extract was combined with the organic layer. The toluene solution was washed with 35% hydrochloric acid, water to a pH of 6-7, 5% aqueous ammonia and water until neutral. Then the solution was clarified with activated carbon and evaporated, removing the remaining toluene with methanol. The evaporation residue (0.323 g) contained a mixture of compounds III, VI and 17α, 21-dihydroxy-11β-trifluoroacetyloxy-6-methylene-pregn-4-en-3,20-dione in a ratio of 26: 57: 17% (HPLC) . Column chromatography on SiO _{2 was} used to separate the mixture and identify compounds.

17α, 21-dihydroxy-11β-trifluoroacetyloxy-6-methylene-pregn-4-en-3,20-dione, so pl. 120-122 ° C. ¹ H NMR (400 MHz, CDCl ₃ , δ, ppm): 5.87 (1H, s, H-4), 5.67 (1H, q, 11-H, J = 2.8 Hz), 5.10 (1H, t 6-C = CH ₂ , J = 1.9), 5.01 (1H, t, 6-C = CH ₂ , J = 1.9), 4.27 dd, 4.64 dd (AB, 2H, 21-CH ₂ , J = 4.5, J = 19.9), 1.17 (3H, s, H-19), 0.80 (3H, s, H-18).

B. Scheme, option 2: III-IV- [V] -VI

Example 6. Obtaining 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6-methylene-pregn-4-en-20-one (VI)

Option 1

To the reaction mixture containing 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6ξ- (N-methyl-N-phenylaminomethyl) -pregna-4-en-3.20-dione (V), obtained from 0.5 g (0.972 mmol) 3-methoxy-21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy pregn-3,5-dien-20-one (IVa) according to variant 1 of the synthesis scheme (example 4), 0.38 ml (0.446 g) was added 35 % hydrochloric acid (0.156 g of hydrogen chloride, 4.283 mmol). The reaction mass was stirred at room temperature for 30 minutes, then diluted with 10 ml of methyl alcohol and poured into 20 ml of water. The precipitate was filtered off, washed on the filter with water to a pH of ~ 7 wash water, and dried to constant weight. Recrystallization from methanol gave 0.31 g (0.605 mmol) of compound VI with a yield of 62.24%, counting from IVa. Mp 187-190 ° C.

Option 2

To a solution of 0.215 ml (0.212 g, 1.98 mmol) of N-methylaniline in 1.25 ml of tetrahydrofuran was added 13 mg (0.068 mmol) of p-TSC monohydrate and 0.2 ml (0.218 g, 0.08 g in 100% calculation , 2.666 mmol) of a 37% formaldehyde solution. Then, with stirring and room temperature, a solution of 0.504 g (0.98 mmol) of 3-methoxy-21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-pregna-3,5-dien-20-one (IVa) in 2.5 was added ml of tetrahydrofuran. The reaction mass was stirred for 1 h at room temperature, then 20 ml of toluene and 0.38 ml (0.446 g) of 35% hydrochloric acid (0.156 g of hydrogen chloride, 4.283 mmol) were added. The reaction mass was stirred at room temperature for 30 minutes, and transferred to a separatory funnel. The acid layer was separated, the toluene solution was washed with 5% hydrochloric acid, water to a pH of 6-7, 1% aqueous ammonia and water until neutral. Then the solution was clarified with activated carbon and evaporated, removing the remaining toluene with methanol. 2 ml of methanol and 10 ml of water were added to the residue, the suspension was kept for 30 min at a temperature of 8-10 ° C, the precipitate was filtered off, washed on the filter with water, dried to constant weight. 0.326 g (0.636 mmol) of compound VI was obtained in 64.9% yield, counting from IVa.

Option 3

To 2 g (3,783 mmol) of 3-ethoxy-21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-pregn-3,5-dien-20-one (IVb) in 2 ml of triethylorthoformate was added 0.48 ml (0.474 g, 4.42 mmol) N-methylaniline. Then, a solution of 0.075 g (0.435 mmol) of p-TSC in 0.75 ml of ethanol and 0.41 ml (0.447 g, 0.165 g in 100%, 5.506 mmol) of a 37% formaldehyde solution was added dropwise. The reaction mass was stirred for 1.5 hours at room temperature and diluted with 20 ml of toluene. After this, 10 ml of acetone, 3 g (14.407 mmol) of barium chloride and 3 ml (4.336 g, 2.385 g in 100%, 24, 31 mmol) of 55% sulfuric acid were added. The reaction mass was stirred at room temperature for 1 hour, then worked up as described in Example 5. 1.83 g (3.571 mmol) of compound VI were obtained in 94.4% yield, counting from IVb.

B. Scheme, option 3: III- [IV] -V

Example 7. Obtaining 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6ξ- (N-methyl-N-phenylaminomethyl) pregn-4-en-20-one (V)

To a suspension of 5 g (9.99 mmol) of compound III in 7.5 ml of trialkyl orthoformate was added 0.125 g (0.725 mmol) of p-TSC. The reaction mass was stirred at room temperature for 15-30 minutes. After the completion of the reaction to obtain 3,5-dienol ether (IVa or IVb), 2.15 ml (2.122 g, 19.8 mmol) of N-methylaniline and 2 ml (2.18 g, 0.807 g in 100% calculation) were added to the reaction mass. 26.86 mmol) 37% aqueous formaldehyde solution. The reaction mass was stirred at room temperature for 2-2.5 hours. At the end of the reaction, to isolate a mixture of 6ξ- (N-methyl-N-phenylaminomethyl) epimers, the reaction mass was diluted with 20 ml of alcohol (methanol or 95% ethyl alcohol) and slowly poured into 200 ml of water. The precipitate was filtered off, washed on the filter with water to a pH of ~ 7 wash water, and dried to constant weight. Compound V (isomer mixture) was obtained in quantitative yield.

G. Scheme, option 4: III- [IV] - [V] -VI

Example 8. Obtaining 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6-methylene-pregn-4-en-20-one (VI)

Option 1

To a suspension of 20 g (39.96 mmol) of 21-acetoxy-11-trifluoroacetyloxy-hydrocortisone (III) in 20 ml (17.82 g, 120.24 mmol) of triethylorthoformate was added 0.5 g (2.90 mmol) of p- TSC and stirred in a stream of argon at room temperature for 15 minutes. In this case, the dissolution of the precipitate of compound III and the formation of a gel-like mass of dark green were observed.

At the end of the reaction, the inert gas was stopped and 4.8 ml (4.74 g, 44.212 mmol) of N-methylaniline and 4.1 ml (4.489 g, 1.654 g in 100%, 55.063 mmol) of 37% aqueous were added slowly dropwise. formaldehyde solution. After adding N-methylaniline, a color change of the reaction mixture from dark green to reddish brown was observed. The reaction mass was stirred at room temperature for 1.5 hours

At the end of the aminomethylation reaction, 200 ml of toluene was slowly added to the reaction suspension with vigorous stirring. The reaction mass was stirred for 30 minutes at room temperature. The dissolution of the gel-like precipitate and the separation of the aqueous layer were observed. To the reaction solution was added 50 ml of a 5% aqueous ammonia solution and the mass was stirred for 10 minutes. A change in color of the reaction mass from red to orange-yellow was observed.

The aqueous layer was separated and extracted with toluene three times. The organic layer and toluene extracts were combined and washed with 5% aqueous ammonia and water until neutral.

Thereafter, 150 ml of methyl ethyl ketone, 30 ml (43.36 g, 23.847 g in 100%, 243.15 mmol) of a 55% aqueous solution of sulfuric acid and 10 g () were added to a solution of compound V in toluene (~ 375 ml) with stirring. 90.11 mmol) calcium chloride. The reaction mass was kept at room temperature and vigorous stirring for 1 h. After the deamination reaction was completed, the reaction mass was transferred to a separatory funnel. The aqueous acidic layer, which is a thick red liquid, was separated from the organic layer and extracted with toluene three times.

The organic layer and toluene extracts were combined, washed sequentially with 5 ml of a 55% aqueous solution of sulfuric acid, with water to a pH of ~ 6-7 (in 10 ml portions); 5 ml of 5% aqueous ammonia and water until neutral.

After washing, the toluene solution of compound VI was dried with sodium sulfate and clarified with activated carbon. Then the solvent was evaporated (in vacuum or at atmospheric pressure). The toluene residue was removed by azeotropic distillation with ethanol.

After complete removal of toluene, 100 ml of ethanol was added to the residue in the cube at a temperature of 50-60 ° C and the mass was stirred at the same temperature until completely dissolved. A solution of compound VI in ethanol, cooled to a temperature of 40-45 ° C, was poured in a fine stream into 500 ml of water with vigorous stirring. The suspension was stirred for 30 min at a temperature of 10-15 ° C, then kept without stirring for 2 hours at the same temperature. The precipitate was filtered and washed with water to pH ~ 7 wash water, dried to constant weight.

20.04 g (39.101 mmol) of 21-acetoxy-11-trifluoroacetyloxy-6-methylene-hydrocortisone (VI) were obtained in a yield of 97.85%. Mp 190-192 ° C.

Option 2

To a solution of 1.3 g (2.597 mmol) of 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-pregn-4-en-3.20 dione (III) in 2 ml (1.782 g, 12.024 mmol) of triethyl orthoformate was added mg (98.13 mmol) p-TSC. The reaction mass was kept in a stream of dry nitrogen for 30 min at room temperature and 0.56 ml (0.553 g, 5.16 mmol) of N-methylaniline and 0.52 ml (0.567 g, 0.21 g in 100% calculation) were added. 6.98 mmol) of an aqueous 37% formaldehyde solution. The reaction mass was held for 1.5 hours. Then 1 ml (1.174 g) of 35% hydrochloric acid (0.411 g of hydrogen chloride, 11.27 mmol) was added and stirred for 30 minutes. After that, 50 ml of ethyl acetate was added to the reaction mass, stirred vigorously for 1-2 minutes, transferred to a separatory funnel. The organic layer was separated and washed with water until neutral, the solvent was evaporated in vacuo. The residue was dissolved in 5 ml of methanol. The resulting solution was added dropwise with stirring to 50 ml of water, cooled to a temperature of 5-10 ° C. The suspension was stirred at the same temperature for 15 minutes, the precipitate was filtered off, washed with water to pH ~ 7 wash water, and dried to constant weight. Received 1.28 g of the technical product of compound VI, which was recrystallized from diethyl ether. Received 1.16 g (2.263 mmol) of compound VI with a yield of 87.15%, so pl. 189-191 ° C.

Option 3

To a suspension of 10 g (19.98 mmol) of 21-acetoxy-11-trifluoroacetyloxy hydrocortisone (III) in 15 ml (14.514 g, 136.77 mmol) of trimethylorthoformate was added 0.26 g (1.51 mmol) of p-TSC and stirred in a stream of argon at room temperature for 15 minutes In this case, dissolution of the precipitate of compound III was observed.

At the end of the reaction, the inert gas was stopped and 4.3 ml (4.244 g, 39.607 mmol) of N-methylaniline, 15 ml of toluene and 4.0 ml (4.36 g, 1.613 g in 100% calculation) were slowly added dropwise, 53, 72 mmol) 37% aqueous formaldehyde solution. The reaction mass was stirred at room temperature for 2 hours.

Then, 7.7 ml (9.04 g, 3.164 g of hydrogen chloride, 86.778 mmol) of 35% hydrochloric acid were added to the reaction solution, and the mass was stirred vigorously for 30 minutes.

At the end of the deamination reaction, the reaction mass was transferred to a separatory funnel. The aqueous layer was separated and extracted with toluene three times. The organic layer and toluene extracts were combined and washed with water to a pH of ~ 6-7 portions of 10-15 ml, 5 ml of 0.5% aqueous ammonia solution, and then with water until neutral.

After washing, the toluene solution of compound VI was dried with sodium sulfate and clarified with activated carbon. Then, the solvent was evaporated in vacuo, removing the toluene residue with methanol.

After complete removal of toluene, 20 ml of methanol and 100 ml of water were added to the residue in the flask with vigorous stirring. The suspension was stirred for 30 min at a temperature of 10-15 ° C, then kept without stirring for 2 hours at the same temperature. The precipitate was filtered and washed with water to pH ~ 7 wash water, dried to constant weight.

8.28 g (16.156 mmol) of 21-acetoxy-11-trifluoroacetyloxy-6-methylene-hydrocortisone (VI) were obtained in a yield of 80.86%.

Option 4

To a suspension of 1 g (1.998 mmol) of 21-acetoxy-11-trifluoroacetyloxy-hydrocortisone (III) in 1 ml (0.891 g, 6.012 mmol) of triethyl orthoformate was added 5 ml of toluene and 0.026 g (0.151 mmol) of p-TSC and mixed under argon flow at room temperature for 15 minutes

At the end of the reaction, the inert gas was stopped and 0.43 ml (0.424 g, 3.961 mmol) of N-methylaniline and 0.4 ml (0.436 g, 0.161 g in 100%, 5.372 mmol) of a 37% aqueous solution were slowly added dropwise. formaldehyde. The reaction mass was stirred at room temperature for 2 hours.

Then, 0.77 ml (0.904 g, 0.316 g of hydrogen chloride, 8.678 mmol) of 35% hydrochloric acid was added to the reaction solution, and the mass was stirred vigorously for 30 minutes.

At the end of the deamination reaction, the reaction mass was diluted with 5 ml of toluene and transferred to a separatory funnel. The aqueous layer was separated and extracted with toluene three times. The organic layer and toluene extracts were combined and washed with water to a pH of ~ 6-7 three times in 5 ml portions, 1 ml of a 0.1% aqueous ammonia solution, and then in a 5 ml three times portions of water until neutral.

After washing, the toluene solution of compound VI was dried with sodium sulfate and clarified with activated carbon. Then, the solvent was evaporated in vacuo, removing the toluene residue with methanol.

After complete removal of toluene, 2.5 ml of methanol and 25 ml of water were added to the residue in the flask with vigorous stirring. The suspension was stirred for 30 min at a temperature of 10-15 ° C, then kept without stirring for 2 hours at the same temperature. The precipitate was filtered and washed with water to pH ~ 7 wash water, dried to constant weight.

0.84 g (1.639 mmol) of 21-acetoxy-11-trifluoroacetyloxy-6-methylene-hydrocortisone (VI) was obtained in a yield of 82.03%.

Option 5

To a suspension of 3 g (5.994 mmol) of 21-acetoxy-11-trifluoroacetyloxy hydrocortisone (III) in 3 ml (2.903 g, 1.804 mmol) of trimethylorthoformate was added 3 ml of toluene and 0.075 g (0.436 mmol) of p-TSC and mixed under argon flow at room temperature for 15 minutes

At the end of the reaction, the inert gas was stopped and 1.29 ml (1.272 g, 11.883 mmol) of N-methylaniline, 1.2 ml (1.308 g, 0.483 g in 100%, 16.116 mmol) of a 37% aqueous formaldehyde solution were slowly added dropwise. and 15 ml of toluene. The reaction mass was intensively stirred at room temperature for 2 hours

Then, 2.31 ml (2.712 g, 0.948 g of hydrogen chloride, 26.034 mmol) of 35% hydrochloric acid 3 g of lithium chloride (70.034 mmol) was added to the reaction solution, and the mass was intensively stirred for 30 minutes.

At the end of the deamination reaction, the reaction mass was treated in the same way as in option 4.

2.78 g (5.424 mmol) of 21-acetoxy-11-trifluoroacetyloxy-6-methylene-hydrocortisone (VI) were obtained in a yield of 90.48%.

Example 9. Obtaining 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6-methylpregna-4,6-diene-3,20-dione (VII)

To a solution of 1 g (1.951 mmol) of 21-acetoxy-11-trifluoroacetyloxy-6-methylenehydrocortisone (VI) in 25 ml of 96% ethanol was added 0.05 g of catalyst (Pd / C or PdO / C) and 0.12 g of anhydrous sodium acetate. The mixture was heated to boiling and 0.1 ml of cyclohexene was added. The reaction mass was boiled with stirring for 0.5 h. The reaction was monitored by UV spectroscopy in a ratio of D ₂₈₆ / D ₂₅₈ , which should be at least 2.6-3.0, and also in a ratio of D ₂₈₆ / D ₂₃₉ , which should be not less than 3.5-4.0. With this ratio of optical densities, the impurities of compounds VI and VIII should not exceed 1 and 2%, respectively.

At the end of isomerization, the hot suspension was filtered, the catalyst slurry was washed with 5 ml of hot ethanol in two stages. The reaction mass was slowly added dropwise to 300 ml of water, cooled to a temperature of 0-2 ° C. The precipitate was filtered off, washed with water until neutral, dried to constant weight.

0.942 g (1.834 mmol) of 21-acetoxy-11-trifluoroacetyloxy-6-methyl-pregna-4,6-diene-3,20-dione (VII) was obtained in a yield of 94.2%. T. pl. 219-221 ° C (decomp.). ¹ H NMR (400 MHz, CDCl ₃ , δ, ppm): 6.01 (1H, br.s, H-7), 5.85 (1H, s, H-4), 5.65 (1H, m, H- 11), 5.07 (1H, d, J = 17.4 Hz, H-21), 4.70 (1H, d, J = 17.4 Hz, H-21), 2.16 (3H, s, CH ₃ C (O)), 1.87 (3H, s, 6-CH ₃ ), 1.20 (3H, s, H-19), 0.86 (3H, s, H-18). ¹³ C NMR (100.6 MHz, CDCl ₃ , δ, ppm): 204.9, 199.4, 170.6, 163.9, 137.3, 131.5, 120.6, 115.7, 88.8, 74.9, 67.7, 52.0, 49.0, 47.8, 35.9, 35.2, 34.6, 33.6, 33.5, 33.2, 23.0, 20.5, 19.9, 18.7, 16.1.

Example 10. Obtaining 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6α-methylpregn-4-en-20-one (VIII)

Option 1

To a solution of 15.1 g (29.463 mmol) of 21-acetoxy-11-trifluoroacetyloxy-6-methylenehydrocortisone (VI) in a mixture of 150 ml of 96% ethanol and 150 ml of acetic acid was added 3.02 g of catalyst (Pd / C or PdO / C ) The mixture was heated to boiling and 45 ml of cyclohexene was added through a dropping funnel over 5 minutes. The reaction mass was boiled with stirring for 1.5-2 hours. The reaction was monitored by UV spectroscopy at a ratio of D ₂₃₉ / D ₂₈₆ , which should be at least 10 (Perkin Elmer Lambda 40 spectrophotometer). With this ratio of optical densities, the impurity of compound VII should not exceed 1%.

At the end of the reaction, the hot suspension was filtered, the catalyst slurry was washed on the filter with hot ethanol (40-50 ° C) in two doses of 5 ml each. To the filtrate was added 0.2 ml (0.235 g, 0.0823 g of hydrogen chloride, 2.254 mmol) of 35% hydrochloric acid to carry out the isomerization reaction of the 6β-methyl group of compound VIIIa into 6α-methyl. The reaction mass was stirred at room temperature for 30 minutes. The content of the 6β-isomer was monitored by HPLC. At the end of isomerization, the reaction mass was evaporated in vacuo on a rotary evaporator to a residual volume of ~ 80 ml. The residue, which is a solution of the product in acetic acid, was slowly added dropwise to 800 ml of water, cooled to a temperature of 0-2 ° C. The precipitate was filtered off, washed with water to pH 7, dried to constant weight.

Received 14.52 g (28.22 mmol) of 21-acetoxy-11-trifluoroacetyloxy-6α-methylhydrocortisone (VIII) in 95.78% yield, and so pl. 113-114 ° C. ¹ H NMR (400 MHz, CDCl ₃ , δ, ppm): 5.74 (1H, s, H-4), 5.67 (1H, m, H-11), 5.06 (1H, d, J = 17.4 Hz , H-21), 4.67 (1H, d, J = 17.4 Hz, H-21), 2.15 (3H, s, CH ₃ C (O)), 1.24 (3H, s, H-19), 1.07 (3H d, J = 6.3 Hz, 6-СН ₃ ), 0.81 (3Н, s, Н-18).

Option 2

Under the conditions of option 1, from 500 mg (0.976 mmol) of 21-acetoxy-11-trifluoroacetyloxy-6-methyl-pregna-4,6-diene-3,20-dione (VII), 460 mg (0.894 mmol) was obtained in 91.6% yield. Mp 112-114 ° C.

Example 11. Obtaining 17α, 21-dihydroxy-11β-trifluoroacetyloxy-6α-methylpregn-4-en-3,20-dione (11-trifluoroacetate 6α-methylhydrocortisone, IX).

Option 1

To a solution of 2 g (3.902 mmol) of 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6-methylene-pregn-4-en-3,20-dione (VI) in a mixture of 20 ml of ethanol and 20 ml of acetic acid was added with stirring 0 4 g of PdO / C. The suspension was heated to boiling point and 6 ml of cyclohexene was added dropwise over 20 minutes. After adding cyclohexene, the reaction mass was boiled for 4 hours. After the completion of the reaction, the catalyst was filtered off, washed with 4 ml of warm ethanol (40-50 ° С) on a filter. The solvent was evaporated to a volume of ~ 4 ml. Then, the residue was added dropwise to 30 ml of cold water. The precipitate was filtered off, washed with water until neutral. The resulting wet cake of a mixture of isomers (VIII and VIIIa) was dissolved in 80 ml of methanol and 0.8 ml of 35% hydrochloric acid was added. The reaction mass was kept at a temperature of 50-60 ° C for 2 hours. At the end of the reaction, the reaction solution was diluted with 80 ml of water. The suspension was kept at a temperature of 10 ° C for 2 hours. The precipitate was filtered off, washed with water to pH 7, and dried to constant weight. After purification, 1.564 g (3.31 mmol) of 11-trifluoroacetyloxy-6α-methylhydrocortisone (IX) was obtained in 84.8% yield, and so pl. 130-132 ° C. ¹ H NMR (400 MHz, CDCl ₃ , δ, ppm): 5.74 (1H, d, J = 1.5 Hz, H-4), 5.67 (1H, m, H-11), 4.63 (1H, dd , J = 4.5, 19.9 Hz, H-21), 4.23 (1H, dd, J = 4.5, 19.9 Hz, H-21), 1.24 (3H, s, H-19), 1.07 (3H, d, J = 6.4 Hz, 6-CH ₃ ), 0.80 (3H, s, H-18).

Option 2

To a solution of 0.1 g (0.194 mmol) of 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxy-6α-methylpregn-4-ene-3,20-dione (VIII) in 2 ml of methanol was added 0.5 ml of water and 0 1 ml (0.117 g, 0.041 hydrogen chloride, 1.127 mmol) 35% hydrochloric acid. The reaction mixture was stirred at room temperature. At the end of the reaction, 10 ml of water was added. The suspension was kept at a temperature of 10 ° C for 2 hours. The precipitate was filtered off, washed with water to pH 7, and dried to constant weight. Received 0.07 g (0.148 mmol) of compound IX with a yield of 76.3%.

Example 12. Obtaining 6α-methylhydrocortisone (I)

Option 1

To a solution of 2 g (3.887 mmol) of 21-acetoxy-11-trifluoroacetyloxy-6α-methylhydrocortisone (VIII) in 10 ml of methanol in a stream of nitrogen, 2 ml (2.218 g, 0.222 g of NaOH, 5.545 mmol) of a 10% aqueous sodium hydroxide solution were slowly added. at room temperature for 30-40 minutes, periodically monitoring the pH of the reaction mass (pH should be no more than 8). The reaction mass was kept at room temperature in a stream of nitrogen for 15 minutes.

At the end of the reaction, the reaction mass was neutralized with a 5% hydrochloric acid solution and 20 ml of a saturated aqueous solution of sodium chloride was added. The reaction mass was stirred for 5 minutes, then 50 ml of ethyl acetate was added, stirred vigorously for 1-2 minutes and transferred to a separatory funnel. The organic layer was separated, the aqueous was extracted twice with ethyl acetate. The combined extracts (~ 100 ml) were washed twice with 5 ml of a saturated solution of sodium chloride and then with water until neutral. Then the solution was treated with activated carbon. The solvent was evaporated in vacuo to stop the run. To the residue was added 20 ml of diethyl ether. The suspension was boiled for 30 minutes, cooled to room temperature. The precipitate was filtered off, washed on the filter with 10 ml of ether, dried to constant weight. 1.253 g (3.328 mmol) of 6α-methylhydrocortisone (I) were obtained in 85.6% yield. Mp 204-206 ° C. ¹ H NMR (400 MHz, DMSO-d ₆ , δ, ppm): 5.49 (1H, s, H-4), 5.16 (1H, s, 17-OH), 4.64 (1H, t, J = 5.6 Hz, 21-OH), 4.49 (1H, dd, J = 5.1, 19.3 Hz, H-21), 4.28 (1H, br.s, 11-OH), 4.24 (1H, m, H-11) 4.06 (1H, dd, J = 5.1, 19.3 Hz, H-21), 1.33 (3H, s, H-19), 0.97 (3H, d, J = 6.4, 6-CH ₃ ), 0.74 (3H, s, H-18).

Option 2

Under option 1, from 1 g (2.116 mmol) of 11-trifluoroacetyloxy-6α-methylhydrocortisone (IX), 0.69 g (1.833 mmol) of 6α-methylhydrocortisone (I) was obtained in 86.6% yield. Mp 203-205 ° C.

Claims (41)

1. The method of obtaining 6α-methylhydrocortisone (11β, 17α, 21-trihydroxy-6α-methylpregn-4-en-3,20-dione) or its esters of General formula I

where R = COCF ₃ or H; R ₁ = COSH ₃ or H, from 21-hydrocortisone acetate of formula II

the sequence of transformations, including the protection of the 11β-hydroxyl group by esterification with a trifluoroacetic acid derivative in an aprotic solvent under basic catalysis with the formation of 21-acetoxy-17α-hydroxy-11β-trifluoroacetyloxypregn-4-en-3,20-dione of the formula III

Eno-esterification of the Δ ⁴ -3-ketosystem of the obtained compound by the action of trialkyl orthoformate in the presence of an acid catalyst with the formation of the 3-alkoxy ester Δ ^3,5 -3-hydroxy derivative of the general formula IV

where R = CH ₃ or CH ₂ CH _3, its condensation with Mannich’s reagent in the presence of an acid catalyst with the formation of a mixture of 6α- and 6β-isomers of 6- (N-methyl-N-phenyl) -aminomethyl derivative of the general formula V

cleavage of the resulting mixture of Mannich steroid bases via a C-N bond with a mineral acid in the presence of hydrohalic acid salts to form a 6-methylene derivative of formula VI

isomerization of the 6-exomethylene bond into the 6,7-endomethylene bond to form compound VII

catalytic hydrogenation of the 6,7-endomethylene bond to form a mixture of 6α-methyl and 6β-methyl isomers and the subsequent isomerization of the 6β-methyl isomer to the 6α-methyl derivative of formula VIII

removal of the protective groups of 11β- and 21-hydroxyl groups in whole or in part. 2. The method according to p. 1, characterized in that for the esterification of the 11β-hydroxyl group of compound II, trifluoroacetic acid anhydride is used as the trifluoroacetic acid derivative. 3. The method according to p. 1, characterized in that the esterification of the 11β-hydroxyl group of compound II is carried out in an aprotic solvent, which is used dialkyl ketones, or cyclic ethers, or dimethyl sulfoxide under basic catalysis using a catalyst in the amount necessary for complete conversions of the starting compound. 4. The method according to p. 3, characterized in that the dialkyl ketone is selected from the group comprising acetone, methyl ethyl ketone. 5. The method according to p. 3, characterized in that the cyclic ethers are selected from the group comprising tetrahydrofuran, dioxane. 6. The method according to p. 1, characterized in that dimethylaminopyridine is used as an esterification catalyst. 7. The method according to claim 1, characterized in that the enolization of the Δ ⁴ -3-keto system of compound III for esterification with the formation of the 3-alkoxy ester Δ ^3,5 -3-hydroxy derivative of the general formula IV is carried out by the action of protic acids. 8. The method according to p. 1, characterized in that sulfonic acids are used as organic acids for the enolization of the Δ ⁴ -3-ketosystem of compound III. 9. The method according to p. 8, characterized in that the sulfonic acids are selected from the group comprising sulfosalicylic acid, p-toluenesulfonic acid. 10. The method according to p. 1, characterized in that the trialkylorthoformate to form the 3-alkoxy ester Δ ^3,5 -3-hydroxy derivative of General formula IV is taken in an amount sufficient to carry out the reaction to obtain formulas IV and V. 11. The method according to p. 1, characterized in that the formation of the 3-alkoxy ester Δ ^3,5 -3-hydroxy derivative of General formula IV is carried out in a proton or aprotic solvent, which is used aliphatic alcohols, cyclic ethers, aromatic hydrocarbons of a series of benzene or mixtures thereof. 12. The method according to p. 1, characterized in that methanol or ethanol is used as aliphatic alcohols. 13. The method according to p. 1, characterized in that as cyclic ethers use dioxane or tetrahydrofuran. 14. The method according to claim 1, characterized in that benzene or toluene is used as aromatic hydrocarbons. 15. The method according to p. 1, characterized in that as a trialkyl orthoformate use trimethyl orthoformate or triethyl orthoformate. 16. The method according to p. 1, characterized in that the 6-aminomethylation of the compounds of general formula IV is carried out in a protic or aprotic solvent, which is used aliphatic alcohols, cyclic ethers, aromatic hydrocarbons of a number of benzene or mixtures thereof. 17. The method according to p. 1, characterized in that the 6-aminomethylation of the compounds of general formula IV is carried out in a trialkyl orthoformate medium or in a mixture of trialkyl orthoformate and an organic solvent without isolating the compound of general formula IV from the reaction mass. 18. The method according to p. 1, characterized in that the deamination of a mixture of compounds of General formula V is carried out in an aprotic solvent or a mixture of aprotic solvents by the action of dilute sulfuric acid using an effective amount thereof. 19. The method according to p. 1, characterized in that as the salts of hydrohalic acid use salts of alkali or alkaline-earth metals, or ammonium, and as hydrohalic acid use hydrochloric or hydrobromic acid. 20. The method according to p. 1, characterized in that the isomerization of the 6-exomethylene bond of the compound of formula VI with the formation of the 6,7-endomethylene bond of compound VII is carried out using palladium or palladium oxide adsorbed on carbon as a catalyst. 21. The method according to p. 1, characterized in that the hydrogenation of the 6.7-endomethylene bond of compound VII is carried out using palladium or palladium oxide adsorbed on carbon as a catalyst. 22. The method according to p. 1, characterized in that the removal of the protective group of the 21-hydroxyl group is carried out by the method of acid solvolysis, both with or without isolation of the hydrogenation product of formula VIII from the reaction mass to form 11β-trifluoroacetate 6α-methylhydrocortisone IX (R = COCF ₃ , R ₁ = H). 23. The method according to p. 1, characterized in that the deamination of the mixture of compounds of General formula V is carried out without isolating them from the reaction mass.