Патент USA US2866809код для вставки
ice 2,866,799 I Patented Dec. 30, 1958 v 2 and may have one or several nuclear double bonds. The 2,866,799 number, nature or location ofsuch nuclear substituents METHQD ‘FOR QQNVERTZNG A 17-20,20-21 BIS or double bonds does not affect or interfere with the above process for forming the dihydroxyacetone side METHYLENEDTGXY OR A 17-20,20-21 BIS-SUB STITUTED METHYLENEDIOXY STEROID OF THE PREGNANE SERIES TO A 17a,21-DIHY-. DROXY-Zll-KETU STEROID OF THE PREGNANE SERIES ' chain since the rest'of the molecule does not-participate . in the reaction. As would be expected, any‘ nuclear substituents subject to acid hydrolysis will be removed - but this is not a fundamental molecular change and in most cases is incidental to obtainment of a particular Roger E. Beyier, Westiield, and Lewis H. Sarett, Prince ton, N. 5., assignors t0 Merck & Co, Inc, Rahway, type of steroid. . ; ’ ' - ' - Serial No. 632,026 As will be‘ seen- from the partial structural formula above, the starting materials for our process are steroidal compounds having twohydrogen atoms or one hydrogen atom and one lower alkyl radical attached’ to each of 12 Claims; (Cl. 260-39145) two carbon atoms of the bis-dioxy moiety. Such prod-’ . As’ described and claimed in our copending patent ap aldehyde. Alternatively, a l7-20,20-21 bisethylidenedioxy (n=l), 17-_20,20-‘2l bispropionaldioxy’ (12:2), 17-20, N. 5., a corporation’ of New Jersey _ No Drawing. Application January 2, 1957 nets are made by treating a 17a,21-dihydroxy-20.-keto steroid ‘of the pregnane series with an aldehyde in the This invention relates to a process for making steroid presenceof a strong acid. The nature of these substitu compounds. More particularly, it relates to a’ process ents depends, therefore, on the particular aldehyde with vfor elaborating a dihydroxyacetone side chain attached to a steroid nucleus. Still more particularly it is con 20 which the ‘1704,21-dihYd1'QXY-20-k6t0 moiety has been in activated. ‘Thus, we may use, and prefer to use, a cerned with a method for converting a 17-20,20-21 bis 17-20,20-21 bismethylenedioxy steroid in which the value methylenedioxy or a 17-20,20-21 bis-substituted methyl enedioxy steroid of the pregnane series to a 17a,2l of n in the formula above is O, and which is obtained by reaction of the 17a,2l-dihydroxy-ZO-ketone with form dihydroxy~20~keto steroid of the pregnane series. plications Serial Nos. 632,009 and 632,010, ?led Jan uary 2, 1957, the dihydroxyacetonemoiety of a steroid ' 20-21 bisbutyraldioxy (1t=3), 17-20,20-21 bisvaleral- ' dioxy (n=4),, ‘or a 17-'20,20-21 biscaproaldioxy (n=5) compound may be inactivated or “blocked” by treatment of the .eroid witha suitable aldehyde whereby a 17-20, _ 20-21v bismethylenedioxy or 17-20,20-21 bis-substituted seroid compound of the pregnane series may be obtained methylenedioxy steroid derivative is formed. Such de with acetaldehyde, propionaldehyde, butyraldehyde, valet‘? aldehyde or caproaldehyde, and employed satisfactorily rivatives are valuable since they permit processes such as starting material for the'process of this inventionQ. as a Claisen condensation or a Grignard reaction to be‘ Typical examples of 17-20,2‘0-,21 bismethylenedioxy and 17,20,20-2Ibis-substituted methylenedioxy steroids which carried out on the steroid nucleus without undesirable involvement of the dihydroxyacetone side chain. This side chain is normally at carbon atom 17 of the nucleus and for this reason the discussion of our invention‘ is directed to such steroids. It may, of course, be located ” are converted by our process to l7a,21-dihydroxy-20-l<eto steroids are: 17-20,20-2l-bismethylenedioxy-pregnan-3-o1 17- 4,20-21-bismethylenedioxy-4-pregnen-3-one at other positions of the nucleus such as at C-16 or C-17a of a D-homo steroid, and the process of our in 17-20,20_2l-bismethylenedioxy-4épregnene-3,1 ledion'e 17- 20,20 - 21 - bismethylenedioxy'- 1,4 - pregnadiene vention is equally applicable to such types of steroids. 3,11-dione It is a purpose of the present invention to provide a process for reforming the dihydroxyacetone side chain from the l7-20,20-21 bismethylenedioxy or 17-20,20-21 bis-substituted methylenedioxy steroid. It is a more spe 45 ci?c object to provide a method of treating a 17-20,20-21 _ y , 17 - 20,20 - 21 - bisethylidenedioxy 3,1l-dione 1,4 - pregnadiene . ' t 17 - 20,20 - 21 - bisbutyralidoxy - 9a, - chloro - 4 '- preg nene-11?-ol-3-one. 20—21 bismethylenedioxy (or bis-substituted methylene ' ' v 17 -20,20 - 21 - bisvaleraldioxy - 90c - bromo - 4 - preg dioxy) steroid of the pregnane series with acid, thus forming a 17ot,21-dihydroxy-20-keto-pregnane. Further objects will be evident from the discussion of this proc ' I l7-20,20-21-bisbutyralidioxy-4-pregnene-3,ll-dione and a 17ot,21-d1hYd1'OXY-20-k€t0 steroid obtained. A still more de?nite object is a method of treating a 17-20, . ‘ l7-20,20-21-bisethylidenedioxy-4-pregnene-1 1,8-ol-3-one . steroid with acid whereby the bisdioxy group is removed hereinbelow. - 17-20-20-21-bisethylidenedioxy-4epregnene-3 ,1 l-dione bisrnethylelnedioxy (or bis-substituted methylenedioxy) ess Y 17 - 20, 20 - 21 - bismethylenedioxy - 1,4 - pregnadiene 1 1/9-ol-3-one ' nene-11?-ol-3-one ‘ . ~ " , . - 17 - 20,20 - 21 - bismethylenedioxy - 9m - ?uoro - 1,4 .- pregnadiene-ll?-bl-S-one , - - 17 - 20,20 - 21 - bismethylenedioxy - 12oz - ?uoro - 1,4 Our process may be pictured structurally as follows: pregnadiene-3,1 l-dione - , ‘ 17,20,20 - 21 - bispropionaldioxy - 4 - pregnene - 6 - ol 3,l1-di0ne ' ' ‘ I 6'01 17 - 20,20 - 21 - bismethylenedioxy - 6 - meth .pregnadiene-l 1/8-ol-3-one ' 1 - 1,4! - 17 - 20, 20 - 21 - bismethylenedioxy - 12a - chloro - 4 C-0 pregnene-l l?-ol-B-one OH I ~ . _ 17 '- 20,20 - 21 - bismethylenedioxy - 4,6 - pregnadiene - (StI’) st () 65 3,11-dione \ - ‘ 17~ - 20,20- 21 - bismethylenedioxy - 9oz - ?uoro - 4 — preg~ In the above equation n is a whole integer having a value of O to 5 inclusive. The expression St'is intended to 17 - 20,20 - 21 - bismethylenedioxy - 4 ~ pregnen - 9a,1l? mean the cyclopentanopolyhydrophenanthrene nucleus diol-3-one _ l with the exception of carbon atom 17 which is separately 17 -20,20-2 1-bismethylenedioxy-alloprcgnane3, 1 l-dione shown. The steroid condensed ring nucleus may be 70 17 - 20,20 - 21 - bismethylenedioxy - 19 -,nor - 4 - preg— substituted'at one or several of the nuclear carbon atoms nene-11?-ol-3-one , v ; . ‘ 2,8 66, 799 d 4 . The 17a,21-dihydroxy-20-keto moiety; either as such l7-20,20-21-bismethylenedioxy~4-pregnen-6-ol-3-one ' or esteri?ed at positions 17 or 2]., is an integral part of 17 - 20,20 - 21 - bismethylenedioxy - 4,9(11) - pregnadien 3-one ‘ the several steroids which have pronounced cortisone-like ‘ activity. 17 ‘- 20,20 '- 21 - bismethyle‘nedioxyw 9,11 .~ oxide -. 4 preg‘nen-S-one‘“ . i 17 - 20,20 .-. 211 - bis 3,1l-dione. ‘ ‘ has been inactivated or “blocked” as a 17-20,20-21 bis methylenedioxy -, 1,4‘ .- tpreg'nadiene . ‘ ‘ ‘ ‘ methylenedioxy-or l7-20,20‘2l bis-substituted methylene dioxy derivative. ‘ 17 - "0,20 . 121 - bism‘ethylenedioxy -1:4 - pregame 5111s- 1 ol-3-one ‘ . The new process described herein is a ready method of reforming this part of the molecule after it “ The following examples are given for purposes of 10 illustration and not by way of limitation: . l7,20,20-21¢biscaproaldioxyépregnane-Z‘s,1lldione , EXAMPLE 1 l7 - 20,20 ‘- 21 ~ bispropionaldioxy- 2 1''‘ methyl - 4 - lpreg nene-3,ll-dione . . 1 . 1. ' - 17 - 20,20 - 21 - bisethylidenedioxy - 4 - pre'gnen- 9:2,11‘5 diol-3-one 1 ‘ 1 . 1 . . ‘In ‘carrying four‘ process ‘afsfo‘lution ‘or suspension of the 1712020121 ismet‘hyle‘ri ‘io‘x'yor'17-‘20,20-2l bis substiu‘ited methylenedioxy steroid of the pregnane series Cortisone acetate To 100 mg. of l7-20,20-21-bismethylenedioxy-4-preg 15 nene-3,l1-dione is added 5 ml. of methanol and 5 ml. of 5 N sulfuric acid and the heterogeneous mixture re ?uxed on the steam bath for one hour (homogeneous after 45 minutes). The methanol is removed in vacuo is ‘treatednwith ‘acid and preferably with a strong ‘acid. We ‘prefer ‘to 'use ‘perchloric acid, formic acid, ‘acetic acid or ‘mineral acids ‘such ‘as hydrochloric, hydrobromic, ‘sul furic ‘fdrup‘hos'phoric acids, and ‘the like. The reaction and the resulting aqueous solution ‘thoroughly extracted 20 withethyl acetate. ‘ is preferably brought about in a "one-‘pliase‘solvent system. When “an ‘organic )acid iscmplo'yed to ‘bring about re generation or ‘tile dihyd‘r‘oxyacelton‘e side chain, we ?nd it suitable and convenient ‘to use ‘an excess of ‘the ‘acid asfthels'olyent‘imedium forj‘the'reaction. For example, in ‘the, conversion of‘ the‘ ‘17-‘20,20-21 bism‘ethylenedioxy derivativesof,cortisone and “hydro‘cb'rtisone ‘to the parent 1 ‘ebmpauaas‘ws have‘use'd aceticandformic acids as both 30 The extract is Washed with sodium bicarbonate, dried and concentrated to give 96 mg. of residue. This 1is dissolved in 0.3 ml. of pyridine and 0.3 ml. ‘of acetic ‘anhydride. The 'mixture is heated for ten minutes ‘on the steam bath and poured into water. The resultant mixture is extracted several times with meth ylene chloride, the methylene chloride washed with dilute hydrochloric acid and sodium bicarbonate, dried over magnesium sulfate and evaporated. The residue is re crystallized from acetone to .give cortisone acetate. solvent "and ‘acid. Alternatively, a lower aliphatic 'car EXAMPLE 2 boiylic, acidfs‘ii'eh "as ‘acetic ‘o'r propionic ‘acid, can serve as‘ thefisoly‘e ‘ass a ‘stronger acid employedrt‘o bring about ‘th'e‘r’hydrolysis. Low'er alkanols are ‘another class Hydrocortisone acetate To 690 mg. ‘of 17-20,20-21-bismethylenedioxy-lt-preg ne‘ne-115-ol-3-one is added 50 ml. of 50% acetic acid. of ‘satisfa‘c‘t‘ory’solvent's‘ for 'the process. “Likewise. a two This mixture is heated under nitrogen ‘at 100° C. for 61/3 hours. The reaction ‘mixture is then concentrated under vacuum‘to dryness. The residue is dissolved in 2.0 ml. p‘hase, tench “system ‘may be ‘utilized, with the steroid dissolv‘d illa ,iiit‘able water-immiscible organic solvent such "a; "chloroform, ‘methylene ‘chloride, benzene 'and the like‘. The”oi‘ga‘riic phase and ‘aqueous acidic ‘I‘pha'se‘ are mixed by agitation to effect the reaction. ‘ ‘ p The reaction ‘times‘and temperatures ‘are not critical, 410 and optimum ‘conditions‘will ‘vary with ‘the particular steroidYs‘ta‘rt‘in'g‘.‘material and ‘acidic reagent employed. Generally, we obtain optimum ‘results at ‘temperatures aqueous sodium bicarbonate. Drying and evaporation of methylene chloride gives'742 mg. ‘of crude hydro cortisonej acetate. Chromatography on alumina yields substantially ‘pure hydrocortisone acetate in the ether chloroforin (3 to 7, 2 to 8, and 1 to 9) e?luents. rangingfrom about 15f’ C. up to about 100"’ C.,‘ and at*‘reacti‘o‘n‘1times1 ‘of about 30 minutes to about ‘50 hours. We prefer to carry out our process at about 25-75“ C. for about three totenhou‘rs. ‘ . of pyridine and 1.8 ml. of acetic anhydride and kept at room‘t'emperature for 18‘ hours. After pouring into water the mixture is extracted several times with methylene chloride. The combined methylene chloride extracts are 'w'asherl‘with 2.5 N hydrochloric acid and saturated ‘ In the same manner, and using 540 mg. of 17-20,.20 With any particular 17720,20-2l bismethylenedioxy (or bis-substituted methylenedioxy st‘e'roid)‘or ‘acidic reagent Zl-bismethylenedioxy-l,4-pregnadiene - 3,11 - dione as starting material, prednisone acetate is obtained. the cou‘rse'of the reaction and ‘the ‘best operating condi tions can be ‘easilyand ‘readilylde'te'i-‘mined by an assay EXAMPLE 3 with jdianisole ‘ bisdiphenylt'etrazolium"chloride, otherwise Cortisone acetate known as the RT. reagent. With this assay,‘ a 1711,21 dihydroxyHZO-ke'to ‘steroid"will1'give with ‘the B. T. re agent and under the conditions describéd‘tby‘Mader ‘et al., Anal. Chem, 24, 666 ‘(19521) "a 'color. The intensity of the color, which is easily ‘measurable quahtita~ ‘ tively against a standard “solution, 'is a measure ‘of the amount of 1711,21-dihydroxy120-ltetolsteroid‘ present 1 since the -l7-20,'20-2¢11bisdioxy ‘s't'artingl‘r‘n'aterials of ‘our process do not give a positive test in ‘the B. T, assay. “ ‘ At “the‘ completion 'ef1our-1prec'eg's, ‘the formed‘?all 60 200mg. of 17-20,20-21-bismethylenedioxy-4-pregnene~ 3,1‘l-dione in 2.0 ml. of 198-100% formic acid is heated at 80° ‘C. for 20’ minutes. After cooling it is poured into about 10 ml. of water and extracted several times with methylene chloride. The combined solvent extracts are washed with saturated sodium bicarbonate solution, dried and concentrated to yield 203 mg. of semicrystalline prod uct. Hydrolysis of this product with sodium methoxide dihydroxy-ZO-keto steroid is isolated ‘ibylany or thef‘méth in methanol, followed by acetylation of the hydrolysis product with acetic anhydride-pyridine yields cortisone ods known to. those skilled‘ in the "steroid ‘art. - For‘ex acetate. ample, the reaction solvent may‘ belremovedby‘ concen tration and the steroid'ektrhote'd Pinto‘ a water-immiscible organic solvent. The solvent is then removed 1and the desired ‘compound obtained lineuhs't'antially pu‘re‘for'm from the residue by chromatography on alumina‘n- ‘We prefer to form a C-21 ester of ‘the 'steroid‘with a lower aliphatic carboxylic acid after removal of the‘reaction solvent but ‘prior to any puri?cation steps. TThese 21 esters,<and particularly the‘acetates,1’are»‘highly crystalline compoundsand more readily puri?ed lthan theYfree-zl alcohols. . When a mixture of 150 mg. of 17-20,20-21-bismeth ylenedioxy-1,4-pregnadiene-1l?~ol-3-one is heated at 75° C. for 30 minutes, and the mixture then worked up as described above, substantially pure prcdnisolone ace tate is obtained. EXAMPLE 4 p l Hydrocortisone To ‘ 50 ‘mg. "of -1‘7-20,20-21-bismethylenedioxy-4-preg~ nene—ll?-ol-3~one in 2.0 ml. of glacial acetic acid is 2,866,799 added 9.2 mluof 60% perchloric acid. The mixture is‘ kept at room temperature for 11 hours. It is poured into .water and extracted'sevcralptimes with ethyl acetate. The combined ethyl acetate extracts are washed,,with saturated sodium, bicarbonate and concentrated to give 6 form‘solutioniis dried and concentrated to dryness‘ in vacuo. The residual 4,9(1l)-pregnadien-17a,21-diol-3,20r diQnejZ‘I-acetate may,’ if desired, be further puri?edby chromatographing on alumina. 46 mg. of crude hydrocortisone. EXAMPLE 10 EXAMPLE 5 l 7. .nllegregrtah-ijazz-dialog1,20-rri0ne ZI-acetate 9ay-?uoro-cgrtisone Following the procedure of Example 9 with 500 vmg. To 100 mg. of 17-20,20-21-bismethylenedioxy-9a-?u~ 10 of 17-20,20-21-bispropionaldioxy-allopregnane-3,lvl-dione, oro-4-pregnene-3,11-dione in 2.5 ml. of glacial acetic acid there is obtained from the chloroform extract allopregnan is added 0.1 ml. of concentrated hydrochloric acid. After ~ 17a,21-diol-3,11,20-trione 2l-acetate. 42 hours at room temperature the reaction is worked up In the above examples, the starting materials may be as described in Example 4 to give 9a-tluorocortisone. prepared by treating a chloroform solution or suspension 15 of a 17a,21-dihydroxy-20-keto steroid appropriately sub EXAMPLE 6 stituted or unsaturated in the nucleus with formaldehyde Cortisone acetate in the presence of hydrochloric acid to make the 17-20,20 To 100 mg. of 17-20,20-21-bismethylenedioxy-4 21bismethylenedioxy compounds, or with a lower alkyl pregnene-3,ll-dione in 2.5 ml. glacial acetic acid is added aldehyde in the presence of hydrochloric acid to make the 0.2 ml. of concentrated sulfuric acid. After 14 hours 17-20,20-21 bis-substituted methylenedioxy compounds. at room temperature the mixture is treated as in Example The reaction is conveniently conducted at about 30° C. 4. _A residue is obtained which on further puri?cation ‘ for ?ve hours although for optimum yields the formation yields substantially pure cortisone. of bisdioxy compound is followed by means of the B. T. test referred to above. At the end of the reaction period, EXAMPLE 7 the two layers are separated, and the aqueous layer ex 6-dehydr0cortisone acetate tracted with fresh chloroform. The chloroform solutions are combined, washed with 5% sodium bicarbonate and A mixture of 200 mg. of 17-20,20-2l-bismethylene dried. The chloroform is then removed and the residual dioxy-4-6-pregnadiene-3,1l-dione, 12 ml. of methanol and 12 ml. of 5 N sulfuric acid is re?uxed for 90 minutes. 30 bisdioxy steroid further puri?ed, if desired, by recrystal- . lization or chromatographic methods well recognized in The alcohol is removed by concentration in vacuo, and the art. ‘ ’ the residual solution extrated with three 10 mL-portions Although the above examples are given in order to of ethyl acetate. The organic extracts are combined, teach speci?cally and to illustrate methods of carrying washed with dilute sodium bicarbonate and dried over magnesium sulfate. The solvent is removed in vacuo 35 out our invention, it will be evident that'ordinarily re actions or transformations are carried out in the nucleus leaving a residue which is dissolved in a mixture of 1 ml. of pyridine ‘and 1 ml. of acetic anhydride. After heat ing on the steam bath for 10 minutes, the mixture is poured into water, and the aqueous phase extracted with of the 17-20,20-21 bisdioxy steroid before such compounds are treated by our process to re-form the dihydroxyacetone side chain. What is claimed is: three '5 ml.-portions of chloroform. The chloroform 40 1. The process which comprises treating a steroid of solutions are washed with water, acid and base, and dried the pregnane series having at carbon atoms 17, 20 and 21 over magnesium sulfate. On evaporation of the solvent, the structure ‘and crystallization of the residue from acetone, 4-6 pregnadien-l7a,2l-diol-3,l1,20-trione 2il-acetate is ob tained. ' EXAMPLE 8 6-m‘ethyl prednisone acetate 180 mg. of 17-20,20-21-bismethylenedioxy-6-methy-1, 4-pregnadiene-3,ll-dione in 3 ml. of, glacial acetic acid is treated with 0.5 ml. of concentrated sulfuric acid, and the reaction mixture held at 35 ° C. for 10 hours. The mixture is then poured into water and extracted with three small portions of methylene chloride. The organic extracts are combined, washed with dilute sodium bicar bonate and the solvent removed in vacuo. The residue is dissolved in pyridine~acetic anhydride as described in Example 2. The 6-methyl prednisone acetate thus ob tained is isolated by the procedure set forth in Example 1. EXAMPLE 9 4,9 (11 ) -pregnadien-1 70:,21 -di0l-3,20-di0ne 21 -acelaz‘e' 55 wherein n is a whole integer having a value of 0 to 5 in clusive with an acid selected from the class consisting of perehloric, formic, acetic and mineral acids thereby form ing a l7a,2l-dihydroxy-ZO-keto steroid of the pregnane series. 60 2. 3. 4. 5. The process The process The process The process ' of claim of claim of claim of claim 1 1 1 1 wherein n is 0. wherein n is 1. wherein n is 2. wherein n is 3. 6. The process which comprises treating a 17-20,20-21 bismethylenedioxy steroid of the pregnane series having 5l0 mg. of 17-2020-21 bisethylidenedioxy-4,9(l l) at the 3-position a member of the class consisting of keto pregnadien-3-one is added to 45 ml. of 50% acetic acid and hydroxy substituents with an acid selected from the and the resulting mixture heated on the steam bath under classconsisting of perchloric, formic, acetic and mineral nitrogen for eight hours. The mixture at the ‘end of acids thereby forming a l7a,2l-dihydroxy-20-keto steroid this time is concentrated to dryness in vacuo, and the res of the pregnane series having at the 3-position a member idue dissolved in 3.0 ml. of pyridine and 2.0 ml. of acetic of the class consisting of keto and hydroxy substituents. anyhydride. This solution is held at 40° C. for 12 hours’ 70 7. The process which comprises treating l7-20,20-21 and then poured into 30 ml. of water. bismethylenedioxy-4-pregnene-3,1l-dione with sulfuric The resulting aqueous solution is extracted with three acid thereby forming 4~pregnene-17a,2l-diol-3,l1,20 20 ml.-portions of chloroform, and the combined solvent trione. , extracts washed successively with 2 N hydrochloric acid, 8. The process which comprises treating 17-20,20-21 5% sodium bicarbonate solution and water. The chloro bismethylenedioxy-4-pregnene-11,8-ol-3-one with acetic 9,866,799 8 ‘~11. The processlwhich comprises treating 17-20,20-21 bismethylenedioxy-1,4-pregn1adiene3,11~dione with formic acid thereby forming hpredhisone. 10. The process which comprises treating 17-20,20-21 bisrnethylenedioxy-li-pregnene-11B-ol-3-pne with perchlor ic acid thereby forming ‘4-pregnene-1‘1?,17a;21-tridl-3,20 dione. ‘ t ‘ hydrochloric acid thereby forming 9u~?uorocorti'sone. “12. Whe ‘process which comprises treating -17-20,20-21 =55 bis‘ethylidenedioxy-4;9( 1 1 ) -pregnadien-3-one with acetic acid thereby forming 4,9(1'1)'pregnadien-17a,2l-diol 3,20-dione. No references cited.