JPH0830076B2 - 16-dehydro-20-oxopregnane derivative and process for producing the same - Google Patents

Description

TECHNICAL FIELD The present invention relates to a 16-dehydro-20-oxopregnane derivative and a novel method for producing the same.

[Conventional technology]

Since 1950, 16-dehydro-20-oxopregnane derivatives have been used as intermediates in the synthesis of corticosteroids (PLJulia).
n) Others: Journal of American Chemical Society (J.Am.Chem.Soc.) 72 , 5145 pages (1950)
Year)).

These derivatives include various sapogenins such as diosgenin, ticogenin or hecogenin (R.E.
Marker (REMarker) and others: Journal of American Chemical Society (J.Am.Chem.Soc.) 61 , 359
2 page (1939); ibid. 62, 518 pages (1940); ibid. 6
9, 2167 page (1947)) and aglycone discard Roy Dar alkaloids, for example, solasodine or tomatidine (Wai Sato (Y.Sato) Other: (. J.Org.Chem) Journal of Organic Chemistry 22, 1496 Page (1958)) manufactured by chemical oxidative decomposition.

Since the mid-1960s, plant sterols, especially sitosterols, have replaced sapogenins and steroidal alkaloids as starting materials in the synthesis of drugs because of their low cost and ready availability.

3-oxo-23,24-dinol-4,17 (20) -choradiene-22-acid (U.S. Pat. No. 3,994,933) and 9α-hydroxy-3-oxo-23,24-dinol-4,1.
7 (20) -Coradiene-22-acid (British Patent No. 2,098,218) is produced by the partial side chain degradation by microorganisms of animal or plant sterols.

Attempts have been made to produce pregnane skeleton intermediates with sterol degradation products having a 23,24-dinol-cholan skeleton for the synthesis of corticosteroids.

3-oxo-23,24-dinol-4,17 (20) -choradiene-22-acid was converted to 4,16-pregnadiene-3,20-dione by hydrogen peroxide to give 9α-hydroxy-3-oxo-23. , 24-Zinol-4,17 (20) -choradiene-22-acid was converted to the novel compound 9α-hydroxy-4,16-pregnadiene-3,20dione in the presence of a transition metal in a neutral or alkaline medium. It can be converted in high yield in the presence of oxoacid salts of (tungsten, molybdenum, vanadium, titanium).

This type of conversion, which is not found in any literature, allows the production of 16-dehydro-20-oxopregnane derivatives from natural sterols in high yield, which may be used as intermediates in the synthesis of corticosteroids. can do.

3-oxo-23,24-dinol-4,9 (11), 17 (20) -colatriene-22-acid, which is not described in the literature, is
Prepared from the known 9α-hydroxy-3-oxo-23,24-dinol-4,17 (20) -choradiene-22-acid with the 9α-hydroxy group removed. The double bond at the 9 (11) position of this molecule is characteristic of corticosteroid preparations.
It paved the way for the formation of β-hydroxy, or 11-oxo groups, or halogen substituents into the carbon 9 position, which is known to increase the anti-inflammatory effects of corticosteroids.

The double bond is introduced into 3-oxo-23,24-dinol-4,9 (11), 17 (20) -colatriene-22-acid at the 1-position by dehydrogenation of the microorganism using a simple substance of Arthrobacter. Was done. This is common with corticosteroids.

[Summary of Invention]

A novel method for converting 23,24-dinorcholanic acid unsaturated at the 17 (20) position to a compound having a pregnane skeleton is 3-oxo-23,24-dinol-4,9 (11), 1.
Known 4,9 (1 from 7 (20) -colatriene-22-acid
1), 16 Puregunatorien-3,20-dione (S. Bernstein) S.Bernstein):. Journal of the American Chemical Society (J.Am.Chem.Soc) 81, 4
956 (1959)) and 3-oxo-23,24
-Zinol-1,4,9 (11), 17 (20) -colatetraene-
A novel intermediate in the synthesis of corticosteroids from 22-acids,
That is, 1,4,9 (11), 16-pregnatetraene-3,20-
Applies to the manufacture of Zion.

Cortico, a 16-dehydro-20-oxo-pregnane derivative prepared with 3-oxo-23,24-dinol-4,17 (20) -choradiene-22-acid or a derivative of this acid substituted in the sterane skeleton It is advantageous to protect the 3-oxo group during conversion to steroids. That is, in this case, the epoxide is selectively formed at the 16 double bond, and the 16α, 17α-oxide derivative can be easily converted to the 17α-hydroxy compound with the 17α-hydroxy corticosteroid (P-L).・ PLJulian et al .: Journal of American Chemical Society (J.Am.Chem.Soc.) 72 , 5145 pages (1950)).

Therefore, 3-oxo-23,24-dinol-4,17 (20)
The 3-oxo group of the steroid having a -corradien-22-acid structure is protected by a blocking group (oxime, ketal, enol-ether), and the derivative thus obtained is a 16-dehydro-20-oxopregnane derivative. Is converted to.

The present invention relates to a method for producing a 16-dehydro-20-oxopregnane derivative, wherein a steroid having a 17 (20) -dehydro-23,24-dinorcholan-22-acid structure or salt is a transition metal oxoacid salt. Reacts with hydrogen peroxide in a neutral or alkaline medium in the presence of a catalyst.

In the production method of the present invention, as a starting material, 23,24-dinorcholan-22- having a double bond at the 17 (20) position is used.
It is advisable to use all steroids having an acid structure.

In order to produce an optimal intermediate of 16-dehydro-20-oxopregnane structure for the synthesis of natural corticosteroid hormones and drugs, the 17 (20) -dehydro-23,24-dinorcholan skeleton is used in the production method of the present invention. Prior to use, it is desirable to modify the 17 (20) -dehydro-23,24-dinorchorane skeleton to facilitate the introduction of 17α- and 21-hydroxy groups for corticosteroids. It would be economically advantageous to develop synthetic techniques in which the characteristic structural elements of corticosteroids are incorporated into the skeleton of dinorcholanic acid, which can be inexpensively produced by microbial processes.

According to a preferred embodiment of the invention, initially 20 to 50 mol% of the ammonium or alkali metal salt of the transition metal oxoacid used as catalyst is added, if necessary, with an inorganic or organic base, preferably sodium hydroxide. Is generated,
17 (20) -dehydro 23,24- used as starting material
It is added to an aqueous solution of the salt of dinorcolanic acid, and then a 30% hydrogen peroxide solution is added to the reaction mixture at 5 to 50 molar equivalents.

It is desirable to use ammonium paramolybdate or sodium tungstate as the catalyst.

Unlike the salt of dinorcolanic acid used as starting material, the product with a 16-dehydro-20-oxopregnane skeleton is insoluble in water and precipitates from the reaction mixture.

The product can be purified by recrystallization. The components of the mother liquor can be purified by column chromatography and thin layer chromatography.

The structure of the product produced by the production method of the present invention was confirmed by ultraviolet spectrum analysis, infrared spectrum analysis, ¹ H-NMR spectrum analysis and mass spectrum analysis.

〔Example〕

Next, the present invention will be described in detail based on preferred embodiments. However, this embodiment is not limited to the present invention.

Example 1 9α-Hydroxy-3-oxo-23,24-dinol-4,1
Preparation of 9α-hydroxy-4,16-pregnadiene-3,20-dione from 7 (20) -colladiene-22-acid 1076 mg (3 mmol) 9α-hydroxy-3-oxo-2
3,24-Zinol-4,17 (20) -colladiene-22-acid was dissolved in a mixture of 30 ml water and 3 ml 1N sodium hydroxide. Then 531 mg of ammonium paramolybdenate tetrahydrate was dissolved in 6 ml of water and p
H was adjusted to 8.0 with concentrated ammonium hydroxide and the two solutions were combined.

To the pH 8.5 reaction mixture, 20 ml of 25% hydrogen peroxide was added dropwise at room temperature over 2 1/2 hours. Foaming of the reaction mixture was suppressed by adding a few drops of dichloromethane. After the reaction is complete, the mixture is removed to remove dichloromethane.
It was left at 45 ° C for 30 minutes and then cooled to room temperature. The precipitated product was filtered and dried.

Yield: 650 mg of 9α-hydroxy-4,16-pregnadiene-recrystallized first with acetone and then with benzene.
3,20-dione was obtained.

Melting point: 186-192 ℃ Ultraviolet spectrum analysis (ethanol): λ _max 242nm (ε
= 25,400) Infrared spectrum analysis data (KBr): ν _OH 353
0, ν _{C = O} 1665,1650, ν _{C = C} 1610,1585 cm ^{−1 1} H-NMR spectrum analysis data (CDCl ₃ , δ, TM
S): 6.75m (H-16), 5.90s (H-4), 2.25s (3H-21),
1.30s (3H-19), 0.95s (3H-18) ppm Analysis: Calculated value Experimental value C 76.79% 76.67% H 8.59% 8.54% Example 2 3-oxo-23,24-dinol-4,9 (11 ), 17 (20) −
Preparation of 4,9 (11), 16-pregnatriene-3,20-dione from coratriene-22-acid 1703 mg (5 mmol) 3-oxo-23,24-dinol-4,
9 (11), 17 (20) -colatriene-22-acid was dissolved in a mixture of 50 ml water and 5 ml 1N sodium hydroxide.
Then 885 mg ammonium paramolybdenate tetrahydrate was dissolved in 15 ml water, the pH was adjusted to 7.5 with concentrated ammonium hydroxide and the two solutions were combined. To the reaction mixture at pH 8.0, add 15 ml of 25% aqueous peroxide solution (1
10 mmol) was added dropwise at room temperature over 2 hours. The product is
Within this time precipitation started to occur, then the precipitate was filtered and dried.

Yield: recrystallized first with acetone and then with methanol
975 mg of crude 4,9 (11), 16-pregnatriene-3,20-dione were obtained.

Melting point: 202-205 ° C. [α] _D = + 237 ° (C = 1, chloroform) 3-oxo-23,24-dinol-4,9 as starting material
Preparation of (11), 17 (20) -colatriene-22-acid 1792 mg (5 mmol) of 9α-hydroxy-3-oxo-2
3,24-Dinol-4,17 (20) -choradiene-22-acid was added to 18
It was dissolved in ml of dichloromethane and then 1.08 ml of chlorosulfonic acid was added dropwise to the stirred solution at -5 ° C. The stirring was continued for 30 minutes at this temperature.

Next, 22 ml of water was added to the reaction mixture with stirring,
The layers formed were separated. The solution layer was extracted twice with 10 ml of dichloromethane each time. The dichloromethane extracts were combined and evaporated under reduced pressure. Balance 1.84
g was purified by thin layer chromatography (developing solvent: benzene to acetone 8: 2).

The pure product thus obtained was dissolved in 30 ml of methanol and then 2 ml of 10% methanolic hydrochloric acid was added. The acidic methanol solution was diluted with 120 ml of ethyl acetate and the mixture was washed twice with 25 ml of water each time and then evaporated under reduced pressure.

Yield: 1.44 g of 3-oxo-23, recrystallized from methanol,
24-dinol-4,9 (11), 17 (20) -colatriene-22
An acid is obtained.

Melting point: 238-239 ℃ Ultraviolet spectrum analysis (ethanol): λ _max 236nm Infrared spectrum analysis data (KBr): ν _OH 3400
~ 2400, v _{C = C} 1675 (COOH and 3-keto), v _{C = C}
1610 cm ^{-1 1} H-NMR spectrum analysis data (CDCl ₃ , δ, TM
S): 5.70s (H-4), 5.50m (H-11), 2.00s (3H-2
1), 1.30s (H-19), 0.90s (3H-18) ppm Mass spectrum data: M ^{+ ・} 340, m / z: 340,32
5,307,295,227,213,105,91.

Example 3 Preparation of 4,16-pregnadiene-3,20-dione from 3-oxo-23,24-dinol-4,17 (20) -coradiene-22-acid 1028 mg (3 mmol) of 3-oxo -23,24-dinol-4,
17 (20) -Coradiene-22-acid was added to 30 ml of water and 3 ml of 1N.
Dissolved in a mixture of sodium hydroxide. To this solution was added 160 mg (0.5 mmol) of sodium tungsto dihydrate in 1 ml of water.

Then 4.0 ml of 30% hydrogen peroxide solution was added dropwise to the reaction mixture over 1 hour with stirring at 60 ° C. Vigorous foaming was reduced by the addition of a few drops of methyl ethyl ketone. The mixture was left at 5 ° C. overnight. The precipitate was filtered and dried.

Yield: 623 mg recrystallized with acetone for analysis
4,16-pregnadiene-3,20-dione was obtained.

Melting point: 186 to 189 ° C. [α] _D = + 175 ° (C = 0.9, chloroform) UV spectrum analysis (ethanol): λ _max 241 nm (ε
= 28,100) Data by infrared spectrum analysis (KBr): ν _{C = C} 1
650,1680, ν _{C = C} 1585,1615 cm ⁻¹ analysis: Calculated value Experimental value C 80.72% 80.90% H 9.03% 9.12% [Example 4] 3-oxo-23,24-dinol-1,4,9 ( 11), 17 (20)
-Preparation of 1,4,9 (11), 16-pregnatetraene-3,20-dione from colatetraene-22-acid 1015 mg (3 mmol) 3-oxo-23,24-dinol-1,
30 ml of 4,9 (11), 17 (20) -colatetraene-22-acid
Of water and 3 ml of 1N sodium hydroxide. To this solution was added 160 mg (0.5 mmol) of sodium tungsto dihydrate in 1 ml of water. Then 3.2 ml of 25% hydrogen peroxide solution was added dropwise to the reaction mixture over 1 hour and 30 minutes with stirring at 60 ° C.

After cooling the reaction mixture to room temperature, the formed precipitate was filtered and dried.

First, thin-layer chromatography (adsorbent: Kieselgel G commercially available from REANAL in Budapest, Hungary, and Kieselgel 60H).
A mixture of F _{254 + 366} in a ratio of 2: 1; a developing solvent: a mixture of benzene and acetone in a ratio of 4: 1), and then purified by repeating recrystallization twice with benzene to obtain 534 g of a crude product. 1,
4,9 (11), 16-pregnatetraene-3,20-dione was obtained.

Melting point: 204-208 ℃ Ultraviolet spectrum analysis (ethanol): λ _max 240nm (ε
= 24,300) Data by infrared spectrum analysis (KBr): ν _{C = O} 1
670,1660, ν _{C = C} 1630,1610,1590 cm ^{-1 1} H-NMR spectrum analysis data (CDCl ₃ , δ, TM
S): 7.20 (1H-1), 6.30 (1H-2), 6.05 (1H-4), AMXm
(J ₁ , ₂ = 12Hz, H ₂ , ₄ = 2Hz, J ₁ , ₄ = OHz), 6.
70t (1H-16), 5.60m (1H-11), 2.30s (3H-21), 1.45
s (3H-19), 0.95s (3H-18) ppm Mass spectrum data: M ^{+ ·} 308. m / z: 308,2
93,265,172,157,121,91,43.

Analysis: Calculated value Experimental value C 81.78% 81.70% H 7.84% 7.74% 3-oxo-23,24-dinol-1,4,9 as starting material
Preparation of (11), 17 (20) -Coratetraene-22-acid Single Arthrobacter genus obtained by 4-androstene-3,17-dione to make 3-oxosteroid-Δ ¹ -dehydrogenase enzyme 10 ml of medium,
Aseptically weighed into 10 500 ml Erlenmeyer flasks. Then, each medium was diluted with 90 ml of 1% by weight β-cyclodextrin solution.

100 mg 3-oxo-23,2 dissolved in 2 ml ethanol
4-Zinol-4,9 (11), 17 (20) -colatriene-22
Acid was added to each medium and shaken on a rotary shaker for 2 days at 37 ° C. Then combine the broth and adjust its pH.
Is adjusted to 2 with 2N sulfuric acid, and fermented broth is 200m each time.
It was extracted 3 times with l of ethyl acetate.

The ethyl acetate extracts were combined and evaporated under reduced pressure. 960 mg of the crude product thus obtained were purified by thin layer chromatography in a developing solvent of benzene to acetone of 8: 2.

710 mg of the pure product thus purified, 20 ml
Dissolved in methanol, and then 1 ml of 10% hydrochloric acid at 0 ° C
In methanol at. The acidic solution was diluted with 100 ml of ethyl acetate, then washed twice with 25 ml of water and evaporated under reduced pressure.

The product thus obtained, namely 3-oxo-23,24-dinol-1,4,9 (11), 17 (20) -colatetraene-22-acid, was recrystallized from methanol.

Melting point: 240-244 ℃ Ultraviolet spectrum analysis (ethanol): λ _max 235nm Infrared spectrum analysis data (KBr): ν _OH 3600
~ 2500, ν _{C = C} 1690 (COOH), 1665 (3-keto), ν
_{C = C} 1630, 1610 cm ^{-1 1} H-NMR spectrum analysis data (CDCl ₃ , δ, TM
S): 7.20 (H-1), 6.30 (H-2), 6.10 (H-4), A
MXm (J ₁ , ₂ = 10Hz, J ₂ , ₄ = 2Hz, J ₁ , _{4 to} OHz),
5.50m (H-11), 1.95s (3H-21), 1.40s (3H-19), 0.95s
(3H-18) ppm Mass spectrum data: M ^{+ ·} 338 m / z: 338,323,310,211,165,145,121,91 [Example 5] 3-methoxyimino-23,24-dinol-4,9 (11), 17
Preparation of 3-methoxyimino-4,9 (11), 16-pregnatrien-20-one from (20) -colatriene-22-acid 1109 mg (3 mmol) 3-methoxyimino-23,24-dinol-4, 9 (11), 17 (20) -colatriene-22-acid,
It was dissolved in a mixture consisting of 25 ml water, 3 ml 1N sodium hydroxide, and 3 ml dichloromethane. In this solution,
First, 500 mg (1.5 mmol) of sodium tungsto dihydrate was added in 3 ml of water. Then, 5.5 ml of 2
55% hydrogen peroxide solution with constant stirring
The reaction mixture was added dropwise at 0 ° C. over 2 hours. During this addition, the product foamed vigorously and precipitated.

The precipitate thus produced was filtered and dried. Yield: 967 mg of 3-methoxyimino-4,9 (11), 16-
Pregnatrien-20-one, the E and Z isomers, were obtained.

Data by ultraviolet spectrum analysis (KBr): ν _{C = O} 1
659, ν _{C = C} 1620 cm ⁻¹ analysis: calculated value experimental value C 77.84% 77.61% H 8.61% 8.59% N 4.13% 4.53% 3-methoxyimino-23,24-dinol-4,9 (11 as starting material ), 17 (20) -Colatriene-22-acid 2.04 g (6 mmol) 3-oxo-23,24-dinol-4,9
(11), 17 (20) -colatriene-22-acid and 0.55 g (6.
3 mmol) of methoxyamine hydrochloride at room temperature
Dissolved in 15 ml pyridine over time. The reaction mixture was then poured into 60 ml of water and extracted 3 times with 20 ml of dichloromethane each time.

The combined extracts were evaporated under reduced pressure and traces of residual pyridine were removed by azeotropic distillation first with toluene and then with methanol.

Yield: 2.13 g of 3-methoxyimino-23,24-dinol-
4,9 (11), 17 (20) -colatriene-22-acid was obtained.

[Example 6] 5,9 (11), 16-pregnatriene-3,20-dione-3-
Ethylene ketal 1.920 mg (5 mmol) 3-oxo-23,24-dinol-
Cola-5,9 (11), 17 (20) -triene-22-acid-3-
Ethylene ketal was added with 8.6 ml of methanol, 4.3 ml of methyl ethyl ketone, 5.2 ml of 10M sodium hydroxide, and 86
Dissolved in a mixture of ml water. Add this solution to 4.
In 3 ml of water, 0.21 g of sodium tungstate dihydrate was added.

Then, 12.9 ml of 30% hydrogen peroxide solution was added dropwise at 60 ° C. over 1 hour with constant stirring. The reaction mixture was added to 85
The mixture was heated to ° C, the organic solvent was removed, then gradually cooled, and left at 5 ° C overnight.

Yield: 1.01 recrystallized with methanol for analysis
g (60%) of 5,9 (11), 16-pregnatriene-3,20-
Dione-3-ethylene ketal was obtained.

Melting point: 210-214 ° C Ultraviolet spectrum analysis (ethanol): λ _max 239 nm (ε
= 9600) Data by infrared spectrum analysis (KBr): ν _{C = O} 1
657, ν _{C = C} 1589 cm ^{-1 1} H-NMR spectrum analysis data (CDCl ₃ , δ, TM
S): 5.45m (1H-6, 1H-11), 6.70t (1H-16), 2.25s (3
H-21), 1.20s (3H-19), 0.80 (3H-18) Analysis: Calculated value Experimental value C 77.92% 77.66% H 8.59% 8.29% 3-oxo-23,24-dinol-5 as starting material , 9
Preparation of (11), 17 (20) -colatriene-22-acid-3-ethylene ketal 1.022 mg (3 mmol) 3-oxo-23,24-dinol-
4,9 (11), 17 (20) -Colatriene-22-acid was dissolved in 10 ml of dichloromethane and then 20 mg of p-toluenesulfonic acid, 1 ml of ethylene glycol and 0.5 ml of trimethyl orthoformiate were added. added. The reaction mixture was stirred at room temperature for 4 hours, then heated at 40 ° C. and concentrated to half the volume. After adding a drop of triethylamine, the solution was diluted with dichloromethane, washed with sodium hydrogen carbonate solution and water, dried with sodium sulphate and evaporated under reduced pressure.

The product obtained in almost quantitative yield was recrystallized for analysis.

Melting point: 240-243 ° C Infrared spectrum analysis data (KBr): ν _{C = O} 1
701 cm ^{-1 1} H-NMR spectrum analysis data (CDCl ₃ , δ, TM
S): 11.05 (1H, -COOH), 5.45m (1H-6, H-11), 3.95s
(4H-dioxolane), 1.95s (3H-21), 1.20s (3H-1
9) 、 0.90s (3H-18) ppm Analysis: Calculated value Experimental value C 74.97% 75.02% H 8.39% 8.45%

Front page continuation (72) Inventor Nándor Mack Budapest Hungary 1045 Elsebeto Uzza 19 (72) Inventor Tibor Lang Hungary Budapest 1016 Fenjoe Uzza 11 (72) Inventor Evailkoey Hungary Budapest 1046 Munkyachim Uzza 37 (72) Inventor Rajosh Tordi Budapest Hungary 1118 Varani Uzza 53 (72) Inventor Antonia Ecker Neye Bocany Budapest Hungary 1112 Keroe Uzza 12 (72) Inventor Eva Thord Chardi Hungary Budapest Pashalet Utzah 161 72) Inventor Jura Holwart Budapest, Hungary 1052 Kijo Uzza 2-4 (72) Inventor Karroy Albrecht Budapest 1145 Mexico, Hungary Uttsua 44

Claims (7)

[Claims] 1. A steroid having a 17 (20) -dehydro-23,24-dinorcholan-22-acid structure or a salt thereof,
A process for producing a 16-dehydro-20-oxopregnane derivative, which comprises reacting with hydrogen peroxide in a neutral or alkaline medium in the presence of a transition metal oxoacid salt catalyst. 2. The method for producing a 16-dehydro-20-oxopregnane derivative according to claim 1, wherein ammonium paramolybdenate tetrahydrate is used as a catalyst. 3. A process for producing a 16-dehydro-20-oxopregnane derivative according to claim 1, wherein sodium tungstate dihydrate is used as a catalyst. 4. An α-hydroxy-4,16-pregnadien-3,20-dione. 5. A 1,4,9 (11), 16-pregnatetraene-3,
20-Zeon. 6. 3-Methoxyimino-4,9 (11), 16-pregnatrien-20-one. 7. A 5,9 (11), 16-pregnatriene-3,20-
Dione-3-ethylene ketal.