CH335492A - Process for the preparation of 17-oxy-steroids - Google Patents

Description

  

  Process for the production of 17-oxy-steroids It is already known to use enzymes from adrenal glands, in particular with their homogenates, or even with their flow, hydroxylgrtippeit, and the like. a. in 17a position of. Introduce steroids.

   As interesting as this function of animal enzymes is from a theoretical point of view, it cannot be satisfactory for the preparative and especially industrial production of 17a-oxy-steroids. With microbiological methods, which can also be applied on a large scale, the mentioned reaction has not yet been feasible.



  It has now been found that 17-Ox # - steroids can be prepared using compounds of the Pregnan series, which have a '# hydrogen atom in the 17 position, by aerobic cultures of fungi of the species Leptosphaeria maculans, Cucurbitaria laburni , Lophotrichtts martinii,

          Melanospora parasit.ica and Thielavia terricola produced enzymes act. leaves.



  Starting materials for the new process are generally compounds of the pregnane series, for example pregnanes, allopregnanes, isopregnanes, such as 14- or 17-isopregnanes, and homo- and norpregnanes, for example A-nor-, D-homo- and 19-nor -pregnane, which aufwei sen in the 17-position a hydrogen atom.

   The compounds can be saturated or unsaturated and have any substituents. Double bonds can, for example, be in the 1-, 4-, 5-, 6-, 7-, 9-, 14-, 15- and / or 20-position. The configuration of the starting materials is preferably that of the pregnans, 5a-pregnans, 17a-pregnans or corresponding racemates as obtained in the total synthesis.

    Particularly suitable substituents are free or functionally modified hydroxyl, oxo or carboxyl groups, such as ester, ether, thioester, thioether, thiol and thionester, acetal, mercaptal, ketal, Hydrazone, semicarbazone and enol groups, for example in 3-, 7-, 11-, 12-, 16-, 18-, 19-,

   20 and 2.1 positions. Starting materials are u. a. Cortexon, 9,11- or 11,12-dehydro- cortexon, 16a-oxy-cortexon, 18-oxo- and 18-oxy-cortexon, corticosterone, 11-epi-corticosterone, 11-dehydro-corticosterone, 18 -Oxy-cortico- steron, d1> 4-3,11,20-triketo-21-oxy-pregnadiene, 41.4 - 3,

  20 -diketo -11ss, 21- dioxy-pregnadien, progesterone, 17a-progesterone, 11 = keto-progesterone, 11a- as well as 11 l-oxy-progesterone, 9,11- or 11,12 - dehydro-progesterone, 19 - Oxo-progesterone, 19-nor-progesterone, pregnenolone, 21- Oxy - pregnenolone,

          Aldosterone or its functional derivatives.



  To introduce the 17-oxy group, the starting materials are usually incubated directly with the cultures of the fungi mentioned that grow submerged under aerobic conditions known per se. These are expediently agitated, that is to say shaken or stirred, and contain assimilable carbon, in particular special carbohydrates and, if appropriate, nutrients, for example corn steeple or wort, and inorganic salts.

   Natural, synthetic or semi-synthetic nutrient solutions can therefore be used. The simplest process in practice is described below: The organisms are grown in equipment and under conditions similar to those known in the so-called deep-tank process in antibiotic manufacture. After the cultures have developed, the starting materials mentioned are added in a fine dispersion or solution, for example in methanol, acetone or ethylene glycol, and the incubation continues.

   Finally, the myeel is separated off, the filtrate and / or the mycelial mass is extracted and the reaction products are isolated from the extract in a manner known per se, for example by separation processes, adsorption, chromatography, crystallization, conversion into functional derivatives, such as Girard -Connections and the like.

   The same reactions can, however, also be carried out by first separating the active enzymes from corresponding aerobic cultures of the species mentioned and using them under the exclusion of the growing cultures.



  The products of the process can be used as medicinal products or as intermediate products for their manufacture. Of the greatest practical interest are, for example, cortisone, hydroeortisone, 1,2-dehydrocortisone, 1,2-dehydrocortisone, Reichstein's substance S, 17a-oxy-aldosterone as well as those from progesterone,

       Pregnenolone and 11-ketoprogesterone form 17-oxy derivatives.



  <I> Example 1 </I> 50 parts by volume of sterilized 70 percent wort are inoculated with Leptosphaeria maculans and shaken at 27 for 4 days. To the recently developed culture, a solution of 0.01 part by weight of cortexone in 0.5 part by volume of acetone is added under sterile conditions, and the mixture is further shaken at 27. After 3 days, the mycelium is filtered off and washed a few times with water.

   The culture filtrate is extracted three times with 2'5 parts by volume of ethyl acetate each time. The ethyl acetate solutions are 1-n. Hydrochloric acid, water, 1-n. Washed sodium bicarbonate solution and water, dried and evaporated at 40 to 50 in a vacuum. The residue is chromatographed on 0.2 parts by weight of silica gel, where first with. Chloroform alone, then with.

         Chloroform-acetone mixtures with increasing acetone content is eluted. Impurities containing a small amount of cortexone are eluted with chloroform and chloroform-acetone (9'5: 5). The first chloroform fractions with 10% acetone give a residue on evaporation,

   which, with concentrated sulfuric acid, gives the carmine-red color characteristic of substance S (17a-oxy-cortexon). This residue is recrystallized from acetone, @ ether mixtures, pure substance S from F. =? 02 to 213 "and the specific rotation [.a] D = + 132 (in ethanol) is obtained.



  <I> Example </I> 50 parts by volume of sterilized 70 percent wort are inoculated with Cucurbitaria laburni and shaken at 27 for 5 days. A solution of 0.01 part by weight of cortexone in 0.5 parts by volume of acetone is added to the well-developed culture under sterile conditions, and the mixture is shaken further at 27. After 3 days the Mveel is filtered off and the culture filtrate is extracted as described in Example 1.

   As the paper chromatographic examination shows, the extraction residue mainly contains 17a-oxycortexon (substance S from Reichstein), which is isolated and crystallized as in Example 1, together with some starting material.



  <I> Example 3 </I> 50 parts by volume of sterilized 70 percent wort are inoculated with Lophotriehus martinii and shaken at 27 for 3 days. A solution of 0.01 becomes a well-developed culture. Part by weight of cortexone in 0.5 part by volume of acetone is added under sterile conditions and the mixture is shaken further at 27. After 2 days, the 3lycel is filtered off and the culture filtrate is extracted as described in Example 1.

   The extraction residue contains, as the paper-chromatographic examination shows, together with some starting material mainly 1.7a-oxycortexon (substance S from Reichstein), which is isolated and crystallized as in Example 1. <I> Example </I> 50 parts by volume of sterilized 70 percent wort are inoculated with Melanospora parasitica and shaken at 27 for 3 days.

   A solution of 0.01 part by weight of cortexone in 0.5 volume part of acetone is added to the well-developed culture under sterile conditions and the mixture is further shaken at 27. After 2 days, the Myeel is filtered off and the culture filtrate is extracted as described in Example 1.

   As the paper chromatographic examination shows, the extraction residue contains, together with some starting material, mainly 17α-oxycortexon (substance S from Reichstein), which is isolated and crystallized as in Example 1.



  <I> Example 5 </I> 50 volume parts sterilized 70 percent wort are inoculated with Thielavia terricola and shaken at 27 for 3 days. A solution of 0.01 part by weight of cortexone in 0.5 part by volume of acetone is added to the well-developed culture under sterile conditions and the mixture is shaken further at 27. After 3 days, the mycelium is filtered off and the culture filtrate is extracted as described in Example 1.

   As the paper chromatographic examination shows, the extraction residue mainly contains 17α-oxycortexon (substance S from Reichstein), which is isolated and crystallized as in Example 1, together with some starting material.

 

Claims (1)

PATENT CLAIM A process for the preparation of 17-oxysteroids, characterized in that compounds of the Pregnan series which have a hydrogen atom in the 17 position are obtained by aerobic cultures of fungi of the species Leptosphaeria maculans, Cucurbitaria laburni, Lophotriehus martinii, Melanospora parasitica and Thielavia terricola allows the enzymes produced to act. SUBClaims 1. Method according to patent claim, characterized in that cortexon is used as the pregnan compound. 2. The method according to claim, characterized in that corticosterone is used as the Pregnan compound, 3. A method according to claim, characterized in that 11-dehydrocorticosterone is used as the pregnane compound. 4. The method according to claim, characterized in that d1,4-3,11,20-triketo-21-oxy-pregnadiene is used as the pregnan compound. 5. The method according to claim, characterized in that d 1,4-3,20-diketo-11ss, 21-dioxy-pregnadiene is used as the pregnan compound.