Patent GB-843217-A (GB)
Title: Process for the production of pregnadienes
IPC: C12P33/00
Filed: 1955-08-03
Abstract: <PICT:0843217/IV(b)/1> <PICT:0843217/IV(b)/2> <PICT:0843217/IV(b)/3> 1,4-Pregnadiene compounds having at least the same nuclear and side-chain substituents as cortisone and hydrocortisone and their 9 a -fluoro-, 9 a -chloro- and 9 a -bromo-derivatives and the 21-esters thereof are prepared by a process which includes the step of subjecting a pregnene substituted at the 3-position by a keto, hydroxy or ester group and having a hydroxy or ester group at the 17 a -position, the double bond being in the 4-position when the 3-substituent is ketonic oxygen and in the 5-position when the 3-substituent is a hydroxy or ester group, to the action of an enzymatically active broth produced by growing a micro-organism of the genus Corynebacterium or of the species Bacillus ephaericus or a mutant thereof until a second double bond is introduced at the 1-position and any hydroxy or ester group at the 3-position is replaced by ketonic oxygen with shifting of the 5-double bond to the 4-position.  The preferred micro-organisms of the genus Corynebacterium are C. simplex and C. hongii or mutants thereof.  The principal end products of the above process are 1-dehydrocortisone, 1-dehydrocortisol and epi-1-dehydrocortisol and their 9 a -halogen derivatives and 21-esters thereof. 11-Epi-1-dehydrocortisol may then be oxidised to 1-dehydrocortisone.  These products may be prepared by introducing (apart from the additional double bond in ring A) into a suitable reactant (a) an 11-oxygen function, (b) a 17 a -oxygen function (prior to the dehydrogenation step), (c) a 21-oxygen function, and (d) a 9 a -halogen atom such as fluorine, chlorine, or bromine.  These various functions may be introduced in any order. The dehydrogenation step may be applied to 3-keto-4-pregnenes or 3-hydroxy-5-pregnenes or 3-esters thereof-particularly 3-acyloxy-5-pregnenes.  The dehydrogeantion step hydrolyses 3-ester groups and also oxidises 3-hydroxy groups to form 3-keto-1,4-pregnadienes.  The dehydrogenation step may be brought about using a culture of micro-organisms of the type referred to above using starting compounds which can have hydroxyl, keto, halogen, and ester groups in various positions of the nucleus or side chain; thus, hydroxyl groups may be present in one or more of the 3, 11, 17, 20 and 21 positions. keto groups may occupy one or more of the 3, 11 and 20 positions, and halogen, p  such as fluorine or bromine may be attached to the 9 position.  A 20-hydroxy group is usually oxidised to a 20-keto group during the dehydrogenation process.  Ester groups, particularly those derived from lower alkanoic acids such as acetic acid may be present at one or more of the 3,11,17,20 and 21 positions.  The ester groups may be derived from inorganic and organic acids so that the ester groups may be cyclopentyl and cyclohexyl acetates, propionates and butyrates, phosphate, polyphosphate and sulphate groups.  An ester group at the 21-position may or may not be hydrolysed by the dehydrogenation process.  Ester groups in the 11 b -and 17 a -positions are generally not hydrolysed in the dehydrogenation process.  The hydroxy group in the 11-position of the reactants may be an 11 a - or 11 b - hydroxy group.  By the dehydrogenation step it is possible to convert cortisone into 1-dehydrocortisone, cortisol into 1-dehydrocortisol, 4-pregnene-17 a ,21-diol-3,20-dione into 1,4-pregnadiene-17 a ,21-diol-3,20-dione and 1,4-pregnadiene-17 a ,20b ,21-triol-3-one, 5-pregnene-3b ,20-diol into 1,4-pregnadiene-3,20-dione, 4-pregnene-11b ,21-diol-3,20-dione into 1,4-pregnadiene-11b ,21-diol-3,20-dione, 4-pregnene-21-ol-3,20-dione into 1,4-'pregnadiene-21-ol-3,20-dione, 4-pregnene-21-ol-3,11,20-trione into 1,4-pregnadiene 21-ol-3,11,20-trione, 9 a -fluoro-4-pregnene-11 b ,17 a ,21-triol - 3, 20 - dione into 9 a -fluoro-1,4-pregnadiene-11 b ,17 a ,21-triol -3,20-dione, 5-pregnene-3b ,17 a , 21-triol-20-one 3,21-diacetate into 1,4-pregnadiene-17 a ,21-diol-3,20-dione and 4-pregnene-17 a ,21-diol-d,30-di-one 21-acetate, 4-pregnene-20-ol-3-one into 1,4-pregnadiene-3,20-dione, 5-pregnene-3,17 a ,20,21-tetrol-11-one and its 3- and/or 21-esters into 1,4-pregnadiene-17 a ,21-diol-3,11,20-trione and its 21-ester, and 5-pregnene-3,11 a ,17 a ,20,21-pentol and its 3- and/or 21-esters into 1,4-pregnadiene-11 a ,17 a ,21-triol-3,20-dione and its 21-ester. Other reactants that may be used are 5-pregnene-3,11 b ,17 a ,21-tetrol-20-one and 5-pregnene-3, 17 a ,21-triol-11,20-dione and their 3-acetates and 3,21-diacetates; and the 11 a -hydroxy epimers of the above mentioned 11 b -hydroxy reactants such as 4-pregnene-11 a ,17 a ,21-triol-3,20-dione, 4-pregnene-11 a ,17 a ,20,21-tetrol-3-one, 5-pregnene-3,11 a ,17 a ,21-tetrol-20-one and 5-pregnene-3,11 a ,17 a ,20,21-pentol and their monoand polyesters, such as the 3-acetates, 3,21-diacetates and 3,17 a ,21-triacetates, these 11 a -hydroxy starting compounds yielding 1,4-pregnadiene-11 a ,17 a ,21-triol-3,20-dione or an ester thereof.  The 11-epimers of 1-dehydrocortisol and its esters can be converted into 1-dehydrocortisone and esters thereof by oxidation e.g. with chromic acid with or without pyridine or acetic acid-preferably after esterifying the 21-hydroxy group if free.  The ester group may be subsequently saponified e.g. by refluxing with alcoholic potassium bicarbonate.  To summarize, the various transformations which may be brought about are represented by the equations of Figures 9, 10 and 11.  In Fig. 9, X represents H2, = O, or H (a - or b -OH), Y is -COCH2OH and Z1 is OH or H.  In Fig. 10, W is H, F, CI or Br, X is H2, = O or H(OH), Y is -COCH2OH, -COCH 2OOCR1, or -CHOHCH2OH, Z1 is OH or H, and R1 is an alkyl radical having not more than 8 carbon atoms. In Fig. 11, R is an alkanoyl group having not more than 8 carbon atoms, Y is -COCH2OH, -CHOHCH3, -COCH3OO CR1 or CHOHCH2OH, Z1 is H or OH, and R is an alkyl radical having not more than 8 carbon atoms.  The 11- hydroxy groups may be introduced by the action of a culture of an 11-hydroxylating micro-organism of the type Curvularia lunata which introduces an 11 b -hydroxy groups and of the types Asperillus niger, Rhizopus arrhizus, Rhizopus nigricons and Cunninghomella blakesleena which introduce 11 a -hydroxy groups. 17 a -Hydroxy groups may be introduced by the action of a culture of a micro-organism of the genus Trichothecium and 21-hydroxy groups may be introduced by the action of a culture of a microorganism of the genus Ophiobolus, preferably O. herbatrichus.  Thus using the various processes outlined above the following reaction sequences may be carried out:-  <FORM:0843217/IV(b)/1> <FORM:0843217/IV(b)/2> <FORM:0843217/IV(b)/3> <FORM:0843217/IV(b)/4> In the examples:-(1) 1-dehydrocortisone is prepared from cortisone and cortisol is prepared from cortisol using C. simplex; (2) 1,4-pregnadiene-17 a ,21-diol-3,20-dione and 1,4-pregnadiene-17 a ,20,21-triol-3-one are prepared from 4-pregnene-17 a ,21-diol-3,20-dione using C. simplex; in this example cortisone may be replaced by its 21-ester or 17 a ,21-diester, or by 5-pregnene-3,17 a ,21-triol-11,20-dione 21-acetate, 17 a , 21-diacetate or 3,17 a ,21-triacetate, and cortisol may be replaced by its 21-acetate, 17 a ,21-diacetate or 11b ,17 a ,21-triacetate or by 5-pregnene-3,11b ,17 a ,21-tetrol-20-one 3,21-diacetate, 3,17 a ,21-triacetate or 3,11 b ,17 a ,21-tetraacetate; the latter group of compounds may be replaced by the corresponding 11 a -hydroxy compounds to form the corresponding 11 a -OH dienes; (3) 1,4-pregnadiene-3,20-dione is prepared from 5-pregnene-3 b ,20-diol using C. simplex; (4) 1,4-pregnadiene-17 a ,21-diol-3,20-dione is prepared from 5-pregnene-3 b ,17 a ,21-triol-20-one 3,21-diacetate using C. simplex and by a similar procedure 1,4-pregnadiene-11 b ,21-diol is prepared from 4-pregnene-11 b ,21-diol-3,20-dione, 1,4-pregnadiene-21-o 13,20-diene is prepared from 4-pregnene-21-o 1-3,20-dione, and 1,4-pregnadiene-21-o 1-3,11,20-trione is prepared from 4-pregnene-21-o 1-3,11,20-trione; (5) 1-dehydrocortisol is prepared from cortisol, and from 5-pregnene-3,11 b ,17 a ,21-tetrol-20-one, 5-pregnene-3,11 b ,17 a ,20,21-pentol and 3-acetates thereof using C. hoagii; (6) 1-dehydrocortisone is prepared from cortisone and from 5-pregnene-3,17 a , 21-triol-11,20-dione and 5-pregnene-3,17 a ,20,21-tetrol and 3- acetates thereof; (7) 1,4,6-pregnatriene-17 a , 21-diol-3, 11, 20-trione is prepared from 4,6-pregnadiene-17 a ,21-diol-3,11,20-trione 21-acetate using C. simplex and is then reduced into mild conditions as by means of zinc and acetic and ascorbic acids; this example may be repeated using as reactants other pregnadienes having conjugated double bonds in the A- and B-rings, thus the reactant may contain a 3-hydroxy group in which case 5(6) and 7(8) double bonds are present, on 11 a  or b  hydroxy group a 21- methyl group, or a 21-hydroxy or ester group as exemplified by 4,6-pregnadiene-11 b ,17 a ,21-triol-3,20-dione, and 5,7-pregnadiene-3,11 b ,17 a , 21-tetrol-20-one which can be converted into 1,4,6-pregnatriene-11 b ,17 a ,21-triol-3,20-dione and then hydrogenated to the corresponding 1,4-pregnadiene, and by 4,6-pregnadiene-17 a ,21-diol-3,20-dione which can be converted into 1,4,6-pregnatriene-17 a ,21-diol-3,20-dione and then hydrogenated to the corresponding 1,4-pregnadiene; the 21-ester groups may or may not be hydrolysed by the process; (8) 1,4-pregnadiene-11 a ,17 a ,21-triol-3,20-dione is prepared from 11-epi-cortisol using C. simplex and is converted into its 21-acetate; (9) 1,4-pregnadiene-9a -fluoro-21-o 1 3,11,20-trione and 9 a -fluoro-1,4-pregnadiene-11 b ,21-diol-3,20-dione are prepared from the corresponding 4-pregnenes using C. simplex, 9 a -chloro-1,4-pregnadiene-11 b ,21-diol-3,20-dione is prepared from the corresponding 4-pregnene using C. hoagii and is converted into its 21-propionate, 9 a -bromo-1,4-pregnadiene-11 b ,21-diol-3,20-dione i