DE1216299B - Process for making steroid compounds - Google Patents

Description

FEDERAL REPUBLIC OF GERMANY

GERMAN

PATENT OFFICE

EDITORIAL

Int. Cl .:

Number:
File number:
Registration date:
Display day:

C07c

German class: 12 ο -25/05

A40606IVb / 12o 3rd JuH 1962
May 12, 1966

The present invention relates to a process for the production of 21-phosphorylated steroids of the 16a, 17a-dihydroxy-pregnen- _{un (} j -pregnadiene series.

The preparation of 21-monoesters of 16-hydroxysteroids the Pregnan series has been a difficult problem so far because of any esterification of 16a, 17a, 21-trihydroxysteroids mainly the 16a, 21-diesters and in small amounts 16a- and 21 monoester yielded. The separation and purification of the 21-monoester formed in each case has proven to be proven difficult, time consuming and costly.

It is known that certain 21-monoesters, especially the methanesulfonates and p-toluenesulfonates widely used as intermediates in the manufacture of various pharmaceutically valuable 21-substituted steroids have been found because they easily undergo exchange reactions when using other nucleophilic substances are treated. For example, 21-alkyl (or aryl) sulfonated Steroids easily to the corresponding 21-phosphorylated steroids either directly or via the Intermediate product of the 21-halogen steroids are converted. The accessibility of the 21-phosphorylated Steroids, which are active glucocorticoids, have high water solubility and good stability show therefore depends directly on the accessibility of the corresponding 21-alkyl- (or aryl) -sulfonated Steroids. While 21-alkyl (or aryl) sulfonated derivatives of steroids are only one reactive Have hydroxyl group (C21) can be easily produced by known methods, is the production and isolating the corresponding derivatives of steroids that have more than one reactive Have a hydroxyl group (e.g. Cie and C21), can only be done with great difficulty.

It has now been found that 21-alkyl (or aryl) sulfonated Derivatives of steroids with reactive hydroxyl groups on Cie and C21 in good Yield and can be conveniently prepared when using starting compounds according to the invention of the general formula II. It was also found that these 21-alkyl (or aryl) sulfonated Steroids of the general formula III into their 21-chlorides or iodides by methods known per se and then into the corresponding 21-monophosphorylated Steroids can be transferred as shown below.

The present invention accordingly relates to a process for the preparation of steroid compounds the general formula

Process for making steroid compounds

Applicant:

American Cyanamid Company,

Township of Wayne, N. J. (V. St. A.) Representative:

Dipl.-Chem. Dr. I. Maas and Dr. W. G. Pfeiffer,

Patent Attorneys, Munich 23, Ungererstr. 25th

Named as inventor:

Robert Bruce Brownfield, Spring Valley, N. Y .;

Charles Krieger, Clifton, N. J. (V. St. A.)

Claimed priority:

V. St. v. America 7 July 1961 (122 413) -

OYi

or salts thereof, in which - Ci - C2 - denote the divalent rest

CH ₂ -

CH ₂

or CH

CH ■

X is a hydrogen or fluorine atom, Ri is the group (H, β-ΟΉ), H ₂ or keto oxygen and R ₂ and R _{3 are} lower alkyl groups and at most one of the substituents Yi and Y2 = H by using methods known per se

609 568/577

a) a 21-hydroxysteroid. the general formula

CH ₂ OH

e) and this then with a ketone of the general formula

OR ₅

R ₂

wherein Ri and X have the meaning given above and Rs is a hydrogen atom or a lower alkyl group and Re a lower alkyl group means and

- Ci - C2 - C3 - C4 - C5 - Ce -
the rest

lower alkyl

represents, with a lower alkyl sulfonyl halide or a mononuclear aryl sulfonyl halide converts the compound of the general formula obtained

CH ₂ OSO ₂ R ₂₀

OR ₅

wherein Ri, R ₅ , Re, X and

- Ci - C ₂ - C3 - C4 - C5 - Ce -

have the meaning given above and R ₂ O is a lower alkyl or mononuclear aryl group,

b) hydrolyzed under acidic conditions, the obtained loaJTa-dihydroxy ^ l-sulfonate compound

c) with an alkali iodide or chloride in a polar organic solvent, which obtained 21-iodide or chloride

d) reacts with phosphoric acid and silver phosphate, optionally the 21-phosphate obtained transferred into a salt

\ "

or converts its acetal or ketal.

The 16,17-alkoxymethylenedioxy derivatives of, for example, the following steroids , 16a, 17a, 21-tetrahydroxy-1,4-pregnadiene-3,20-dione; 9a-fluoro-11 / 3,160,170,21-tetrahydroxy- ^ pregnen-S ^ O-dione; 11 / 3,16α, 17α, 21-tetrahydroxy-4-pregnen-3,20-dione; 16α, 17α, 21 - trihydroxy - 4 - pregnene - 3,11,20 - trione; 16α, 17α, 21 - tetrahydroxy - 4 - pregnene - 3,20 - dione; 11 ^, 16α, 17α, 21-tetrahydroxy-1,4-pregnadiene-3.20-dione; 16α, 17α, 21-trihydroxy -1,4-pregnadiene-3,11,20-trione; 9a - fluoro - 16α, 17α, 21 - trihydroxy-1,4 - pregnadiene - 3,11,20 - trione; 9a - fluoro - 16α, 17α,. ¹ I-trihydroxy-1,4-pregnadiene-3, ll, 20-trione and the like. The preparation of the compounds which are identified in the reaction scheme as III and III a, takes place by reacting the compounds II and II a with a lower one Alkylsulfonyl halide or a mononuclear arylsulfonyl halide instead. Such sulfonyl halides include, for example, methanesulfonyl chloride, benzenesulfonyl chloride or p-toluenesulfonyl chloride. This reaction is usually carried out in a solvent that serves as an acid acceptor, for example in pyridine. Optionally, dimethylformamide or dioxane, for example, together with an acid acceptor such as pyridine, triethylamine or the like can be used as the solvent. The reaction is preferably carried out at a temperature in the range of about -20 to 35 ° C. The reaction is usually complete in a few hours, but the time can vary from about 2 to 24 hours. The product is usually insoluble in water and can be obtained by filtration after stirring the final reaction mixture into cold water.

The steroids marked IV on the reaction scheme are prepared from the steroids ΠΙ and III a by subjecting them to acidic hydrolytic conditions in order to remove the vicinal cis-dihydroxy groups in the 16a and 17a positions and (if one of 4-pregnen-3- onen goes out) to restore the zl ⁴ -3-pn-F, unction in ring A.

The hydrolysis can be carried out by adding the steroids III and III a in a solvent such as chloroform or methylene chloride, which contains a catalytic amount of a mineral acid such as concentrated hydrochloric acid, at a temperature in the range from about 25 to 65 ^° C solves. Under these conditions the reaction is complete in about 1 to about 24 hours and, being insoluble in these relatively non-polar solvents, the product can be filtered directly from the reaction mixture or isolated by other conventional methods. The steroids III and III a can optionally be suspended in a solvent such as methanol and treated with an aqueous mineral acid such as dilute hydrochloric acid at a temperature in the range from about 25 to 65 ^° C. for a time from about 1 to about 24 hours.

In practice, the complete hydrolysis is effected by removing the methanolic The reaction mixture was refluxed for a period of about 1 hour. The steroid product IV can from the cooled reaction mixture either directly by filtration or after Dilute the reaction mixture with water. Often times it is not necessary that Purify steroids III and III a, and the crude products become easy hydrolysis conditions subjected to give the final products IV. The following reaction scheme illustrates the method according to the invention.

CH ₂ OH

II

lower alkyl

Ri

CH ₂ OSO ₂ R ₂₀

C = O;, - "0 \ / OR ₆

III

lower alkyl

CH ₂ OSO ₂ R ₂₀

OH

IV

CH ₂ Y

Ri

^x 0H

Va Y = J Vb Y = Cl

45

T / OH
CH ₂ OP (
I ^x O-Na ⁺

Ri

30th

35

40

Ri

55

6o

65 Wherein R _x , R _2a , R ₃ , R ₄ and X have the meaning given above.

There has long been a need for a highly water-soluble form of a 16a-hydroxylated active Steroids, such as triamcinolone, for pharmaceutical preparations. Steroid derivatives such as borate salts and salts of half-esters of steroids (e.g. the 21-sodium hemisuccinates) prepared to obtain such water-soluble forms, however, these forms show a poor one Resistance. On the other hand, it has been found that the 21-phosphate esters of steroids produced, as in Reaction scheme shown from which steroids IV can be made, more stable and as extraordinary water-soluble forms are superior.

The following examples explain the process according to the invention.

example 1

a) Production of 9ct-fluoro-II / S-hydroxyloa.lVa-methoxymethylenedioxy-II-methanesulfonyloxy-1,4-pregnadiene-3,20-dione

9.63 g (22, ImMoI) 9a-fluoro-11) 3.21-dihydroxy-16a, 17a - methoxymethylenedioxy - 1,4 - pregnadiene-3,20-dione are dissolved in 100 ml of dry pyridine

and chilled the solution in an ice-water bath. There are slowly 3.31 g (28.9 mmol) of methanesulfonyl chloride added with stirring to the cold pyridine solution.

The reaction mixture (a turbid solution) is stirred for 1 hour at ice bath temperature and then kept overnight at -1O ⁰ C. The reaction mixture is poured into ice water with stirring, the solid product is removed by filtration, washed with water, 1.4 N hydrochloric acid (50 ml) and finally with water until the wash water is neutral to litmus paper. The air dry product is 11.03 g, mp = 201.5 to 202.5 ⁰ C (decomposition). Cooling the combined mother liquors and wash water gives an additional 0.14 g of solid. The total yield of crude product is 11.17 g (21.7 mmol = 98.2%). An analytically pure sample of 9a-fluoro-11ß-hydroxy-16a, 17a-methoxymethylenedioxy-21-methanesulfonyloxy-1,4-pregnadiene-3,20-dione is obtained in the form of long colorless needles by repeated recrystallization from methanol and has an F. . from 202.5 to 203.5 ° C (decomposition). v _m ax = 1732, 1649, 1605, 1357 and 1174 cm- ^1; [a] f = + 114 ° (c = 1.058 ethylene glycol mono-methyl ether, known under the trade name Methylcellosolve, ldm);
= 238 ταμ, ε = 16 500.

b) Preparation of 9a-fluoro-II / S, 16a, 17a-trihydroxy-21-methanesulfonyloxy-1,4-pregnadiene-3,20-dione

To 10.3 g (20, OmMoI) g
16a, 17a-methoxy-methylenedioxy-21-methanesulfonyloxyl, 4-pregnadiene-3,20-dione, 500 ml of absolute methanol and 20.0 ml of dilute hydrochloric acid are added. The suspension is brought to reflux with stirring, complete dissolution occurring. After about 20 minutes, solid flow begins to separate out, and the amount of solid that separates out during a further 20 minutes at reflux increases continuously (total reflux time 40 minutes). The mixture is cooled overnight (5 ⁰ C) and the solid product was removed by filtration. After washing with cold methanol and drying in air, the product is 8.27 g, melting point = 198.0 to 199.5 ° C. (decomposition). Concentration of the mother liquors gives a further 0.61 g of the product as a colorless solid. The total crude yield is 8.88 g (18.8 mmol) = 94.0%). Repeated recrystallization from methanol gives an analytically pure sample of 9a-fluoro-II ^ -16a, 17a-trihydroxy-21-methanesulfonyloxy-1,4-pregnadiene-3,20-dione in fine, colorless needles with a F. = 197.0 to 197 , 5 ° C (decomposition); v _m ax = 1730, 1660, 1618, 1340 and 1170 cm- ^1; [a] f = + 54.8 ° (c = 0.803, methyl-55 cellosolve, ldm); A ^ _a ^e ° ^H = 239 ηΐμ; ε = 15 100.

c) Preparation of 21-iodine-9a-FluorII / S, 16a, 17a-trihydroxy-1,4-pregnadiene-3,20-dione

6.50 g 13.8 mmol) 9o - FluorII ^, 16a, 17a - trihydroxy - 21 - methanesulfonyloxy - 1,4 - pregnadiene-3,20-dione are suspended in 300 ml of acetone. 3.25 g (21.7 mmol) of sodium iodide are dissolved in 200 ml of acetone, and the solution is added to the steroid suspension with stirring. The mixture is refluxed with stirring for 1 hour and 20 minutes, the suspension becoming considerably thinner, but at no time complete dissolution occurs. The hot mixture is filtered, the solid (water-soluble sodium methanesulfonate) washed with hot acetone and the combined filtrates are cooled overnight. The first crop of long, fine, colorless needles is removed by filtration, washed with cold acetone and finally dried in air, yielding 1.62 g, melting point = 170.5 to 171.0 ⁰ C (decomposition). A second crop of colorless solid is obtained by concentrating the combined mother liquors and wash water. Another 5.26 g of M. = 169 to 170 ^° C. (decomposition) are obtained. The yield is 6.88 g (13.6 mmol = 99.1%). Repeated recrystallization from acetone-petroleum ether (bp = 60 to 70 ⁰ C) gives an analytically pure sample of 21-iodine-9a-fluoro- ΙΙβ, ίβα, Πα - trihydroxy -1,4 - pregnadiene - 3,20 - dione from F. . = 175.0 to 176, ⁰ ° C (decomposition, wherein violet steam at 172 ° C spontaneously fills the tube); vmax = 1695, 1650, 1597, 1068 and 892 cm- ¹ ; [a \ S = + 87 ° (c = 0.586, DMF, ldm); AC & oh = 239 ΐημ, ε = 17 400.

d) Preparation of 9a-fluoro-lljS, 16a, 17a, 21-tetrahydroxy-M-pregnadiene-S ^ O-dione-

21-sodium hydrogen phosphate

15.6 ml of 85% phosphoric acid and 41.5 g of trisilver phosphate are combined with cooling and intimately mixed until a paste forms. 160 ml of acetonitrile are added and, with stirring, to Suspension 25.0 g of 21-iodine-9a-fluoro-II / S, 16a, 17a-trihydroxy -1,4 - pregnadiene - 3.20 - dione and finally a further 160 ml of acetonitrile 'was added. The mixture is refluxed for 4 hours, to room temperature cooled and filtered through diatomaceous earth to remove insoluble solids. Of the The filter cake is washed with methanol, and the combined filtrates are washed in portions with a Sulfonic acid cation exchange resin (H + form) treated until the solution is free of silver ion and exhibits an approximate pH of about 2. The solution slowly becomes weak through a column of a basic ion exchange resin (OH ~ form), which effectively reduces the inorganic Phosphate ions removed from the methanol-acetonitrile solution of the steroid phosphate. The solution is with a 2% solution of sodium methylate brought to pH 5.2 (total volume 2530 ml). the Solution is evaporated to a small volume under reduced pressure, diluted with ether and the suspension obtained is cooled overnight. The product is collected by filtration with Ether washed and air dried, which gives 20.0 g of 9a - fluorine - II / ?, 16a, 17a, 21 - tetrahydroxy-1,4 - pregnadiene - 3.20 - dione - 21 - sodium hydrogen phosphate gives. The qualitative paper strip chromatography NH4OH-water (7: 1: 2) concentrated in the isopropanol-concentrated system shows traces of impurities on non-phosphorylated steroids at Rf = 0.85 and no impurities of inorganic phosphates (Rf = 0.08). The main component (steroid-21-phosphate) appears as a strong spot at Rf = 0.45.

e) Production of 9a-fluoro-II / S, 16a, 17a, 21-tetra-

hydroxy-l ^ -pregnadiene-j ^ O-dione-21-triethylammonium hydrogen phosphate

4.98 g of the above product (impure monosodium salt of '9a - fluoro - II / S, 16a, 17a - 21 - tetra-

hydroxy -1,4 - pregnadiene - 3,20 - dione - 21 - phosphate) are taken up in 70 ml of the lower phase of the system n / 10-NHiOH-n-butanol and extracted with four small portions of the upper phase of the same two-phase system. The aqueous, ammoniacal steroid solution is treated in portions with a sulfonic acid ion exchange resin (H ⁺ form) until the solution has a constant pH of about 2. The solution is then treated in portions with a sulfonic acid ion exchange resin [HN (C2H5) 3+ form] until the solution has a constant pH of 4.2. The solution is treated with nitrogen under reduced pressure to remove dissolved n-butanol, and finally freeze-dried, yielding 3.61 g of the monotriethylammonium salt of 9a-fluoro-11 ^, 16a, 17a, 21-tetrahydroxy-1,4-pregnadiene - 3.20 - dione - 21 - phosphate results as a colorless solid with a melting point of 163 to 170 ^° C. (decomposition); A ™ ^IH = 240 πΐμ; ε = 16,500; [ _a y _D ⁵ = + 49 ° (methanol). Molecular weight (by alkali titration of the free phosphate regenerated in the H + form by sulfonic acid ion exchange resin) = 611 (calculated = 601).

When chromatographing approximately 300 micrograms on paper in the isopropanol-concentrated system NH4OH — water (7: 1: 2) only shows a single spot (Rf = 0.41).

Production of 9a-fluorine-II / S, 16a, 17a, 21-tetra-

hydroxy-1,4-pregnadiene-3,20-dione

21 disodium phosphate

30th

Alternatively, 10 g of the above product (impure monosodium salt of 9a-fluoro-11 / 3,16α, 17α, 21-tetrahydroxy-1,4-pregnadiene-3,20-dione-21-phosphate) in 250 ml of the lower phase of the n / 10-NHiOH - n-butanol system. The solution obtained is extracted with several smaller portions of the upper (butanol) phase of the same system. This treatment effectively removes the unphosphorylated steroid impurities. The pH 9.25 solution is treated with a sulfonic acid ion exchange resin (Na ⁺ form) on the column until the solution shows a constant pH of about 11. The pH of the solution is then lowered to about 6 by treating in portions with a sulfonic acid ion exchange resin (H ⁺ form). At this point the dissolved steroid phosphate exists as a mixture of the mono- and disodium salts that is not contaminated with excess ammonium hydroxide, sodium hydroxide, and non-phosphorylated steroid. The solution is titrated to pH 8.25 (pH of the steroid 21-phosphate disodium salt) with a dilute solution of sodium hydroxide. The solution is treated with nitrogen under reduced pressure to remove dissolved butanol and then freeze-dried, yielding 9.20 g of hydrated 9a-fluoro-II / 3.16a, 17a, 21-tetrahydroxyl, 4-pregnadiene-3.20- Dione-21-disodium phosphate results as a colorless solid. Paper chromatography in the system isopropanol-concentrated NH4OH-water (7: 1: 2) showed a single spot of Rf = 0.45. The solubility of the product in water is over 600 mg / ml.

Alternatively, using the procedure described above and using p-toluenesulfonyl chloride in place of methanesulfonyl chloride, the 9a- fluoro- ilß- hydroxy-16α, 17α-methoxymethylenedioxy-21-ρ-toluenesulfonyloxy-1,4-pregnadiene-3,20-dione obtained in good yield; v _max = 1730, 1660, 1613, 1361 and 1177 cm " ¹ .

The 9a-fluoro-11 ^ -hydroxy-16a, 17a-methoxymethylenedioxy-21-p-toluenesulfonyloxy-1,4-pregnadiene-3,20-dione prepared as above is heated with absolute methanol and dilute hydrochloric acid as described above. The desired 9a - fluoro - ΙΙβ, Ιόα, Πα - trihydroxy-21 - ρ - toluenesulfonyloxy -1,4 - pregnadiene - 3.20 - dione is obtained in good yield with a melting point of 181 to 182 ° C (decomposition), vmax = 1725.1659.1603, mOundmScm- ¹ .

Example 2

a) Production of 3-methoxy-11ß-hydroxy-16a, 17a-methoxymethylenedioxy-21-methane

sulfonyloxy-3,5-pregnadien-20-one

0.36 g of ^ -Methoxy-II / ^ l-dihydroxy-loa ^ a-methoxymethylenedioxy - 3,5 -pregnadien - 20 - one are treated according to the method of Example 1, a) with methanesulfonyl chloride, which gives 0.29 g of 3 -Methoxy-11 (9-hydroxy-16α, 17α-methoxymethylenedioxy-21-methanesulfonyloxy-3,5-pregnadien-20-one gives; _Vmax = 1725, 1650, 1348 and 1170 cm- ¹ .

b) Preparation of ll / ?, 16a, 17a-trihydroxy-21-methanesulfonyloxy-4-pregnen-3,20-dione

3 - Methoxy - ilß - hydroxy - 16α, 17α - methoxymethylendioxy - 21 - methanesulfonyloxy - 3,5 - pregnadien-20-one is treated with dilute hydrochloric acid in absolute methanol according to the method according to Example 1, b), what ll (S, 16a, 17a-trihydroxy-21-methanesulfonyloxy - 4 - pregnen - 3.20 - dione gives; v _ma x = 1722, 1650, 1349 and 1170 cm " ¹ .

Example 3

a) Preparation of 21-chloro-9a-fluorll ^, 16a, 17a-trihydroxy-1,4-pregnadiene-3,20-dione

0.75 g of 9a-Fruor-II & 16a, 17a-trihydroxy-21-methanesulfonyloxy -1,4-pregnadiene-3.20-dione, 202 mg of lithium chloride and 30 ml of dimethylformamide are refluxed for 50 minutes, under reduced pressure to a small amount Concentrated in volume and treated with water. The solid thus obtained is separated by filtration and recrystallized from acetone-petroleum ether (bp. = 60 to 70 ⁰ C) and from isopropanol, giving 278 mg, m.p. = 261-263 ⁰ C (decomposition). An analytical sample is obtained by recrystallizing from methanol, which reduces the melting point to 250-251 ⁰ C (decomposition) λ = 239 πΐμ, ε = 15 300. _Vmax = 3330, 1739, 1678, 1627 and 1610 cm- ¹ ; [a]% ⁵ = + 83 ° (dioxane).

In other experiments it was found that the reflux time was successfully reduced to as little as 8 minutes or that the reaction mixture can simply be stirred at room temperature for 45 minutes.

b) Production of 9a-fluoro-ll ^, 16a, 17a, 21-tetra-

hydroxy-1,4-pregnadiene-3,20-dione

21-sodium hydrogen phosphate

21 - chlorine - 9a - fluorine - ΙΙβ, Ιβα, Πα - trihydroxyl, 4-pregnadiene-3,20-dione is treated with 85% phosphoric acid and trisilver phosphate according to the method of example 1, d), which gives the monosodium salt of 9a - Fluorine - ll / 3.16a, 17a, 21 - tetra-

609 568/577

hydroxy-1,4-pregnadiene-3,20-dione-21 -phosphate yields in a purity that of the product Example 1, d) is comparable. Rf = 0.45 [isopropanol-concentrated NH4OH-water system (7: 1: 2)].

Claims (1)

Claim: Process for the preparation of steroid compounds of the general formula IO ν O / \ -ο / \ / OYi or salts V X T divalent CH ₂ OP ( J OY ₂ ι \ of what C = O rest / R ₂
Γ ^{1 X} Rs ^{R1 =} f
/ ~ 1 V C ₂
I. -Ci-C ₂ ^{o =} \ / 20th - the CH ₂ : CH> 30th CH ₂ or CH X is hydrogen or fluorine, Ri (H, / 3-OH), H ₂ or keto oxygen and R ₂ and R3 are lower alkyl groups, and at most one of the substituents Yi and Y ₂ = H, characterized in that one according to known Methods a) a 21-hydroxysteroid of the general formula CH ₂ OH 45 II 55 wherein Ri and X have the meaning given above and R _{5 is} a hydrogen atom or a lower alkyl group and Re is a "lower alkyl group and - Ci - C ₂ - C3 - C4 · -C ₅ - Ce - ■ the rest CH CH ₂ or / CH ₈ -CH ₂ C C _CH O ^ ^a CH lower alkyl represents, with a lower alkyl sulfonyl halide or a mononuclear aryl sulfonyl halide converts the compound of the general formula obtained CH ₂ OSO ₂ R _2ff wherein Ri, R ₅ , R ₆ , X and
- Ci - C ₂ - C3 - C4 ■ C ₆ - have the meaning given above and R _{2a is} a lower alkyl or a mononuclear aryl group, b) hydrolyzed under acidic conditions, the 16α, 17α-dihydroxy-21-sulfonate compound obtained c) reacts with an alkali iodide or chloride in a polar organic solvent, the 21-iodide or chloride obtained d) reacts with phosphoric acid and silver phosphate, optionally the 21-phosphate obtained converted into a salt e) and this then with a ketone of the general formula R ₂ R ₃ ^ ; c = o or converts its acetal or ketal. 609 568/577 5.66 © Bundesdruckerei Berlin