CH214400A - Procedure for the preparation of keto steroids. - Google Patents

Description

  

  Process for the preparation of ILetosteroids. The dehydration of steroids with, dehydrogenatable hydroxyl groups to the corresponding ketos teroids by chemical means is known. It has now been found that this reaction can also be carried out advantageously by biochemical means.



  The present invention therefore relates to a process for the preparation of keto steroids by dehydrating dehydrogenatable ones. Steroids containing hydroxyl groups, characterized in that a biochemical hydrogen aaceptor absorbs the activated hydrogen from the hydroxyl groups to be dehydrated.



  For the implementation of the process you can bezw the help of certain microorganisms, for example the help of bacteria or mold. the help of enzymes obtainable therefrom. Of the bacteria, for example, the bacteria of the Acetobacter group, in particular the Baeterium (Acetobacter) Pasteurianum, and also bacteria such as Micrococcus oblongus, and sorbose bacteria;

       Of the molds, certain As pergillaceen can be used with advantage.



  The use of so-called "depleted" yeast has proven to be particularly advantageous, as it is available, for example, according to Wieland, "Annalen der Chemie", Vol. 492, p. 183 ff., By shaking yeast suspensions in toluene water with oxygen .



  The biochemical dehydrogenation is expediently carried out under aerobic conditions or in the presence of secondary hydrogen acceptors, such as methylene blue, quinone or the like.



  The steroid compounds which can be used as starting materials for biochemical dehydration include, for example, sterols and bile acids containing dehydratable hydroxyl groups and degradation products of the same, compounds of the cortin and animal type containing dehydratable hydroxyl groups. and herbal heart poisons.

   Furthermore, compounds of the Pregnan series containing dehydrable hydroxyl groups can be used as starting materials, as can compounds of the Oestrau and Androstan series containing dehydrable hydroxyl groups. Of the last-mentioned side chain-free compounds, for example, compounds such as Oes are:

  ertrio, octahydroestrone, and the unsaturated compounds of the androstane series, the androstane compounds dehydroandrosterone and androstend: iol, are specially mentioned.



  If, in addition to the hydroxyl group or groups to be dehydrogenated, the starting material also contains other hydroxyl groups that can be dehydrogenated, then these can, if necessary, be protected from attack by the biochemical dehydrogenating agents.



  <I> Example 1: </I> Unhopped wort for lager beer was used as a nutrient solution for the bacteria. The maltose content of the wort was determined and brought to a content of about 4/10 by dilution with water. After adding calcium carbonate, the nutrient solution was then heated in the autoclave at 115 for 20 minutes and filtered until it had cooled. The nutrient solution, which had been heated again under pressure, was then filled into sterile spatial Petri dishes and inoculated with the Bacterium Pasteurianum.

   After standing for three days at room temperature, a solution of 200 mg of dehydroandrosterone in 15 cm of ethyl alcohol was carefully added to each Petri dish containing 300 cm 'nutrient solution. After standing for three days, it was extracted with ether, the ethereal solution washed, dried and evaporated.

   The residue was then treated for four hours in absolute pyridine with phthalic anhydride to remove the unchanged dehydroandrosterone, and the reaction mixture was poured into water with. Ether extracted and the pyridine removed by washing with acidic water.

    The acidic phthalic acid ester of dehydroandsterone could then be separated off with alkali, and the ethereal solution left behind after evaporation a crystallizate which was treated with acidic ethanol and after recrystallization from hexane androstenedione of the rotation value
EMI0002.0058
    
EMI0002.0059
   and m.p. 173 '.

    <I> Example 2: </I> As described in Example 1, androstenediol monopropionate-17 was subjected to oxidation in a further experiment. namely, 300 mg of this product (mp. 146;
EMI0002.0064
    used for a bacterial culture with 300 cm 'substrate. After standing for four days at room temperature, worked up as described in Example 1, and the unchanged monopropionate was also removed as an acidic phthalic acid ester.

   The testosterone propionate remaining after isomerization was then crystallized from hexane. Tina had the following data:

       M.p. 121;
EMI0002.0075
    
EMI0002.0076
         Example <I> 3: </I> On 1 liter of a nutrient solution of the composition: <B> 1% </B> peptone, 0.2 aö ammonium biphosphate, 0.1 ö potassium biphosphate, 0.025% magnesium phosphate Sorbent bacteria brought to good growth.

    Then 10 cm 'of a 2% solution of cholesterol in alcohol are added to this culture solution in portions. After standing for a long time, the entire reaction mixture is extracted with ether. The ether solution is washed with water, dried and evaporated. The residue. is then reacted in iNlethanol with Semic @ irbazidacetat. Thereby, separation of Cholestenonsemica.rbazon occurs.

   After cleavage, the semicarbazone delivers the cholestenone with the following data: <B> F p. </B> 84-81;
EMI0002.0099
    
EMI0002.0100
   The yield is 29%. <I> Example </I> .l: 8 g of yeast (Milan flaky ferments) are suspended in 30 cm 'of water and mixed with 10 cni \ n / .; Na @ HPO, solution and 10 <B> cm '</B> ni, $ X2P04 solution added. The suspension is shaken in an oxygen atmosphere at 32 for 20 hours;

      190 mg of androstenediol, suspended in 30 cm of water, are then added and the mixture is then shaken in an oxygen atmosphere for a further 47 hours. The reaction mixture is then extracted with ether; the ethereal solution is washed with water, caustic soda, nj, hydrochloric acid and again with water. After drying over sodium sulfate, the ethereal solution is evaporated to dryness.



  The residue is dissolved in 30 cm 'of alcohol, 3 g of glacial acetic acid and 1.5 g of P. Reagent (Girard, Helv. Chem. Acta 19, 1095, 1936) are added and the mixture is heated to boiling under reflux for one hour. After cooling, the whole thing is poured into 140 cm of water, 60 g of ice and the amount of sodium hydroxide necessary to neutralize 2.7 g of glacial acetic acid. It is then extracted three times with ether in order to remove the keto-free portion.



  20 g of 50% strength sulfuric acid are added to the aqueous solution, which causes turbidity. The solution is left to stand for one hour and then extracted three times with ether, the ethereal solution is washed with water, dried over sodium sulfate and the ether is evaporated. If a little ether is added to the residue, it can be obtained in crystalline form.



  After recrystallization of the residue from dilute acetone, a substance of F. 168-169 'is obtained, which proves to be identical to androstenedione. On the basis of the inventive method, it is possible to achieve valuable keto steroids in a relatively simple manner and with relatively high yields, which have hitherto only been relatively cumbersome. were only available in a relatively low yield.

 

Claims (1)

PATENT CLAIM: Process for the preparation of keto steroids by dehydration of dehydratable hydroxyl-containing steroids, characterized in that that a biochemical T @ hydrogen acceptor takes up the activated hydrogen of the hydroxyl groups to be dehydrogenated. SUBClaims 1. Method according to claim, characterized in that one works under aerobic conditions. 2. The method according to claim and dependent claim 1, characterized in that the biochemical dehydration is carried out with the aid of bacteria of the Acetobacter group. 3. The method according to claim and dependent claims 1 and 2, characterized in that the biochemical dehydration is carried out with the aid of Bacterium (Acetobacter) Pasteurianum. 4. The method according to claim and dependent claim 1, characterized in that "depleted" yeast is used to carry out the biochemical dehydration.