DE1946689A1 - A-nor-progesterone, process for their production and their use for the production of pharmaceutical preparations - Google Patents

Description

DR. ELISABETH JUNG ₁ DR. VOLKER VOSSIUS, DIPL.-ING. GERHARD COLDEWEY

PATENT LAWYERS MÖNCHEN 23 · · TELEPHONE 345087 · TELEGRAM ADDRESS: INVENT / MÖNCHEN

. το _{TELEX s 2B βββ}

J5. Sep 1969

UoZoi E 712 (Vo / kä)
Case 760 721 - S.

EoRo SQUIBB & SONS ₉
New York, N ₀ Y ₀ , VoSt.A

^M A-nor-Progesterone, process for their production and their use for the production of medicinal products "

Priority: 18 ₀ September 1968, V.St.A ₀ , No. ₀ 760 721

The invention relates to new A-nor-progesterones of the general formula I.

in which R and R ', which can be identical or different, are hydrogen mean atoms or acyl radicals.

A-nor-progesterone, in which the ll position is & ö? -Oriented, are known from the USAo patent 3 330 861 «,

009814/1899

POST CHECK ACCOUNT: MÖNCHEN 50176 BANK ACCOUNT: DEUTSCHE BANK A.G. MONKS. LEOPOLDSTR. 71. KTO. NB. 80 / 367M

The preferred acyl radicals are derived from hydrocarbon carbonic acids with less than 12 carbon atoms. Examples of these carboxylic acids are the lower saturated aliphatic carboxylic acids, such as acetic acid, propionic acid, butyric acid and terto-pentanecarboxylic acid, the lower olefinically unsaturated fatty acids, the monocyclic aromatic carboxylic acids, such as Benzoic acid and the toluic acids, the lower aliphatic carboxylic acids substituted by a monocyclic aromatic radical, such as phenylacetic acid and ß-phenylpropionic acid, the cycloalkanecarboxylic acids and the cycloalkene carboxylic acids.

The invention also relates to the preparation of the compounds of the general formula I according to a simple and powerful fermentation process The process is characterized by that one 17 Oi-Hydroxy-A-nor-progesterone of the action a microorganism of the genus Curvularia subjectα Das The process runs according to the following reaction equations:

t 3

C = O

B.
R = acyl, R '= acyl

R = ACyI, R ¹ -H 0038U / 1889

It was found that nan a. new intermediate product (7erbind A) of the invention, namely 11ß, 17 Ä-dihydroxy-A-norprogesterone from 17 Qf-Hydroxy-A-nor-progesterone with the help of a β Microorganism of the genus Curvularia or the enzymes of this microorganism under oxidizing and preferably aerobic conditions can produce <>

To produce the connections of the invention, the starting sterol initially exposed to the action of the enzymes of a microorganism of the genus Curvularia under oxidizing conditions This oxidation can best be brought about by using the starting steroid of an aerobic culture of the microorganism incorporated or the parent steroid with air and the enzymes of the non-proliferating cells of the microorganism in aqueous Bringing the medium together

In general, the conditions for cultivating the microorganism for the purposes of the invention (with the exception of the starting compound) are the same as those for the production of antibiotics for other microorganisms. applied werdenο The microorganism is grown aerobically in contact with (into or onto) a suitable culture medium * Efc suitable Hährboden containing sources of carbon and energy · Energy sources come Z ₀ for example, carbohydrates such as molasses, glucose _# maltose, starch, or dextrin, a Fatty acids, fats and / or the initial steroid in embossing »However, the nutrient medium preferably contains a source of assimilable carbon and energy in addition to the parent steroid ₅ mutton ^ ₃ sperm oil,

0098U / 1899

6A0 ORIGINAL

Olive oil, tristearin, tripalnaitin, triolein and trilaurin _o Examples of suitable fatty acids are stearic acid, palmitic acid, oleic acid, linoleic acid and myristic acid »

Suitable sources of assimilable nitrogen include organic substances such as soybean meal, corn steep liquor, yeast extract _r meat extract and / or soluble residues at .Alkohol = · · distillation or synthetic compounds ₉ DOH. simple organic or inorganic compounds such as ammonium salts _p alkali nitrates ρ amino acids or urea, in question ₀

During the fermentation must be a sufficient supply of sterile air to be taken care of ₀ This can be by conventional processes take place, in which one such "B ₀ a large surface area exposed to the culture medium of the air or operates according to the submerged culture with aeration" The starting steroid, the culture during the Incubation time or the nutrient medium prior to sterilization or inoculation. The preferred concentration range of the parent steroid in the culture is about 0.01 to about 0.1 J ^ o The conversion time can vary considerably, it can be from about 24 to about 96 hours. »

The compound A can Z ₀ B ₀ be acylated with a carboxylic acid anhydride, such as acetic anhydride, in the presence of perchloric acid, to give the corresponding 11ß _s 17 0 (-diacyloxy derivative (compound B). This compound can then be with a weak base, ZoB. an alkali metal carbonate, such as potassium carbonate or sodium carbonate, can be saponified to the corresponding llß «-acyloxy-17o (-» hydroxy-A-nor-progesterones,

0098U / 1899

The invention further relates to the use of the A-nor-Erogesterone for the production of medicinal preparations. “ The compounds of the invention are medicaments, they have antiandrogenic activity and they can therefore be used for the treatment of, for example, hyperandrogenic acne.

The concentration and or "or dose depends on the activity of the particular compound used, and the clinical picture from _o In general, about 0.01 mg to about 1.0 / kg body weight, either orally or locally verabfolgte to humans or animals

The examples illustrate the invention.

example 1 HS, 17 Pf -dihydroxy-A-nor-progester on A) fermentation

Curvularia lunata (NRHL-2380) is placed on agar slants using grown on the following nutrient medium (A):

Gram

Glucose 10

Yeast extract 2.5

K ₂ HPO ₄ 1

Agar 20

distilled water to 1 liter

The surface growth of two 2-week-old slant agar tubes is in 5 ml of a 0.01 # aqueous Matriualaurylsulfatlösung suspended. 1 ml portions of this suspension are used to inoculate eight 250 ml Erlenmeyer iolbans used, each 50 ml of the sterilized! H & rmediuma (B) contained"

*. Q098U./1.899.-

- 6 <~ 1346689 Crramai 30th glucose 20th Soybean meal 2.2 Soybean oil 2.5 CaCO ₅ distilled water to 1 liter.

After about 48-stiindiger incubation at 25 ° C on a ureihschüt telmasohine (28C cycles / minute; 5 cm stroke) 10 vol ₀ «$ strength transmissions on Yieruii'-thirty 250-ml Erlemaeyer flask are carried out, each containing 50 ml of the sterilized growing medium (C) contained:

Shark Spring Water 6th HH ₄ H ₂ PO ₄ 3 Yeast extract 2.5 glucose 10 CaCO 2.5 distilled water to 1 liter.

300 micrograms / ml of the starting steroid are then added to each bottle by adding 0.25 πύ. a sterile solution (6ö ing / ml) 17 Oi-Hydroxy-A-nor-progester on. given in 3S ₃ N-DimethylformasaiBl «A total of 510 mg of steroid are fermented«

Another 5 days of incubation under the sliding Bswi® Toretefeenö, öesehrieben ₃ the content $ .S2? Pistons

see W & Ü the Säraaische through ice. liärfll'se ^ fil- I © lfe © ia ₀ aas% cel iiai ö @ s? Filters are successively MQ είΐ Μ% ®Ά (Μ. Warm water ¹ & ewa, sohen _c - Das Ultsfit imd di®

SAD ORIGINAL,

Washless drops are combined and have a volume of 1800 mlc

B) insulation

The filtrate is extracted three times with 600 ml of chloroform each time. The combined chloroform extracts are washed three times with 60C ml of water each time, dried over sodium sulfate and evaporated to dryness Developing solvent is chromatographed. The main band recognizable in UV light is extracted with ethyl acetate. A mixture of products is obtained which is further purified by chromatography on silica gel using ethyl acetate as the developing solvent i> contains methanol, evaporation of the extract, and recrystallization of the Kückstandes from a mixture of methanol and Isopropylather gives the LLSs, 17 tf-dihydroxy-a-nor-progesterone from P ₀ 208 to 2O9 ° C "melts After repeated recrystallization from the same solvent mixture the analysis sample at 210 to 211 ⁰ C;

λ KBr 2.87, 5.88, 5.99 and 6.20ju; χ JgJj ₁ 8.97 (s, 18-CH ₃ ), 8.59 (a, 19-CH ₃ ), 7.72 (s, 21-CH ₃ ), 5.64 (m, 11Of-H) and 4.28 (s, 3-H) o

C H

i 72.26; 8 _S 49;

: 72.27; 8.49 »

009814/1899

OftenGlNAL

Example 2 Ilsstl7 tf-diacetoxy-A-nor-progesterone

A solution of 400 mg of 11β, 17α-dihydroxy-A-nor-progeeterone in 3 ml of acetic anhydride is mixed with a drop of 175% perchloric acid, stirred for 30 minutes at low temperature, then diluted with water and extracted with chloroform. The chloroform extracts are washed with 8 # sodium chloride solution, dried over sodium sulfate and evaporated under reduced pressure. One receives 113, 17tf-diacetoxy-A-nor-progesterone: _{o nTir} ft 9.20 (β, 18-Cn ,; _s o, 77 (8, 19-CH ₉ /, o »01 (8, llß-OAc ), 7.99. (Β, 21-CH ₃ ), 7.93 (s, 17OT-OAc), 4.65 (m, 11 Of-H) and 4.32 (s, 3-H).

Example 3

llß-acetoxy-17 Qt-hydroxy-A-nor-profeesteron

a solution of 323 mg LLSs, 17 'diacetoxy-A-nor-progesterone in 20 ml methanol is treated with 4 ml of a 10 56-aqueous Kaliumcarbonatlösun ^, and the mixture 18 hours at room temperature and stirred ₀ Thereafter, the solution is neutralized with acetic acid and )

evaporated Dex residue is diluted with water and extracted with chloroform ₉ chloroform extracts are washed with 8 $ strength brine, dried over sodium sulfate and eingedampft0 The residue is plattenchromatographiert smittel on silica gel with a mixture of equal parts of chloroform and ethyl acetate as Entwicklungslöeun ^. The main band recognizable in UV light is eluted with ethyl acetate o The ethyl acetate extract is evaporated and recrystallized from a mixture of ethyl acetate and isopropyl ether, yield 113 mg of product with a temperature of 191 to 193 ° Co.

0098 U / 1899

sungBBltt ·! eotuilit die Analyiipxob U% 194 to 196 ⁰ Cι 2.91, 2.97, 5.75, 5.78, 5.88, 5.96 and 6.17 μ%

231 * µ (S «16,100 ); T ^ ₁ 9.13 (β, IS-CHj), 8.79 (β, 19-CH ₃ ), 8.00 (θ, IIβ-OAc), 7.79 (β, 21-CH ₃ ), 4 , 68 (lltf-B), 4.35 (β, 3-Η).

CH C ₂₂ H ₃₀ O ₅ ; ber.ι 70.56; 8.08; Found: 70.57; 8.10.

0098U / 1899

Claims (6)

P ate ntansprtiche l-nor-progesterone of the general formula I. w in which R and R \, which can be the same or different, mean hydrogen atoms or acylate. 2. IIS, 17 flf-Dihydroaty-A-nor-progesteren. 3 · ÜB, 17 α-diaeetoacy-A-nor-progesterone. 4 · llfl-acetaxy-17 CF-hydroxy-A-nor-progeeterone. 5. Terfa & ren «tr production of the compounds according to claim 1, characterized in that nan 17Pf -Hy»> drojqr-A-nor-progesterone subjects the action of a Hikroor ^ auisaua of the genus Curvularia and optionally the obtained llß, 17öf-öihydroxy-A -ior-progesterone acylates a carboxylic acid anhydride in the presence of catalytic amounts of perchloric acid and in addition acylates the resulting llß, 17-tr-diacyl oxy-A-nor-progesterone approximately duroh with an aqueous solution of a weak base in the llB-Aeyioxy-17 Ä ₍ -transferred. 009814/1899 - BAD ORIGINAL 6. The method according to claim 5 »characterized in that aan as a microorganism Curvularia lunata used, 7ο Method according to claim 5 or 6 »characterized in that the carboxylic acid anhydride is used Eesiga acid anhydride used o 0098U / 1899