JPS5910799B2 - Pregnane steroid 17-ester production method - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing pregnane steroid 17-esters, and more particularly to a novel production method that can easily obtain pregnane steroid 17-esters, which are extremely useful as adrenocorticosteroids. It is something.

As a result of various studies, the present inventors succeeded in selectively producing only 17α,21-dihydroxy-20-ketopregnan steroid 17-esters, and completed the present invention.

The present invention will be explained in detail below.

The present invention relates to a partial structural formula (wherein R represents an alkyl group, a cycloalkyl group, or an aryl group, and R' represents a lower alkyl group).

) of a 17α,21-dihydroxy-20-ketopregnan steroid (hereinafter abbreviated as compound σ)) is treated with a hydrolase having activity in an acidic region. .

substructure formula (In the formula, R has the same meaning as above.

) is a method for producing 21-hydroxy-20-ketopregnan steroid 17-esters (hereinafter abbreviated as compound M).

In its implementation, compound (I) is dissolved in alcohol such as ethanol or a mixture of water and alcohol,
1 to 80 parts by weight, preferably 10 to 50 parts by weight, per 1 part by weight of compound (I), is added and suspended in a commercially available hydrolytic enzyme having activity in an acidic region.

At this time, it is not necessary to make the reaction solution particularly acidic by adding an acid.

This is reacted at room temperature for 3 to 4 days, at 30 to 45°C for 10 to 50 hours, preferably at 35 to 38°C for 20 to 40 hours, or by shaking.

After the reaction is completed, the reaction solution is separated, and the P solution is extracted with an appropriate organic solvent such as chloroform or methylene chloride. The extract is washed and dried according to a conventional method, and then concentrated under reduced pressure to obtain the crude product. obtain.

This crude product is recrystallized from an organic solvent such as ethyl acetate or acetone or an organic mixed solvent such as acetone-hexane, acetone-ether, acetone-petroleum ether, etc. to obtain the compound (
II) can be obtained.

Although recrystallization with an appropriate organic solvent is sufficient for purification of the compound ([), means such as preparative thin layer chromatography and column chromatography can also be used if necessary.

The hydrolytic enzyme used in the present invention is obtained by culturing various fungi belonging to the genus Aspergillus, Rhizopus, Trichoderma, etc., and is biodiastase (Amano Pharmaceutical Co., Ltd., originating from Aspergil lusorizae).
, Takajia. :x. Tase (Sankyo Co., Ltd., Aspe
rgillus orizae origin), bernase (Taisho Pharmaceutical Co., Ltd., Aspergillus orizae origin), bernase (Taisho Pharmaceutical Co., Ltd., Aspergillus orizae origin)
Origin), Morsin (Seishin Pharmaceutical Co., Ltd., Aspergil
lus saitoi origin), Sanactase (Meiji Pharmaceutical Co., Ltd., A. lus saitoi origin), Vanprosin (Yakuru Co., Ltd.), Asperg.
Aspergillus niger origin), Denapsin 10p (Nagase Sangyo Co., Ltd., Aspergillus niger origin), Lipase AP (Amano Pharmaceutical Co., Ltd., A sper)
gil lusniger origin), cellulase AP
(Amano Pharmaceutical Co., Ltd., Aspergillus nige
r Origin), Senorerosin AP (Ueda Chemical Industry Co., Ltd.),
A spergi 1 1us niger origin),
Lipase MY (Meito Sangyo Co., Ltd., Candida cy
lin-dracea origin), Sancrose F (Hankyu Kyoei Bussan Co., Ltd., Rhi chinensis origin), Panthera zopus zeus (Yakult Co., Ltd., Trichoderma vi)
Among hydrolytic enzymes that are easily available commercially, such as vepsin obtained from the gastric mucosa of pigs or cows, which have activity in the acidic region. All you have to do is select the appropriate one.

In the present invention, the pH of the reaction solution suitable for the action of these hydrolytic enzymes is 4 to 6, but the pH of the reaction solution is approximately the same as the pH when purified water is stored in the air.
Since it is H, there is no need to add acid to the reaction solution to make it particularly acidic when these hydrolases are allowed to act.

Rather, adding an acid will cause esterification of hydroxy groups other than the 17-position, resulting in a mixture of these undesirable esters in the 17-ester and subsequent purification. Therefore, the purpose of the present invention, which is to generate only 17-ester by a simple procedure, is contradicted.

The 17-esters obtained according to the present invention depend on the starting materials, 17α,21-cyclic orthoesters.

Conventionally, in order to obtain pregnane steroid 17-esters, there is a method of directly esterifying pregnane steroid 17α-hydroxys using an acid. These hydroxy groups must be protected prior to
Furthermore, after esterifying the hydroxyl group at position 17, the protecting groups at positions 11 and 21 must be removed.

Moreover, since a strong acid is used, undesirable side reactions tend to occur, and it is not possible to selectively obtain only the desired 17-ester.

Particularly when a substituent such as a methyl group is present at the 16th position, the reaction does not proceed well.

In addition, pregnane steroids 17α,21-dihydroxys are converted into 17α,21-@-like orthoesters,
There is a method of converting this into 17-esters by acid hydrolysis.

As the acid, either an inorganic acid or an organic acid can be used, but generally weak acids such as oxalic acid or propionic acid are considered suitable.

However, in this case, 21-esters are produced as a by-product in addition to 17-esters due to hydrolysis, and a process for removing unnecessary 21-esters is required.

Hydrochloric acid - sodium citrate, hydrochloric acid potassium hydrogen monophthalate,
Although the production of 21-esters can be reduced to some extent by using an acid-buffered water-organic medium such as hydrochloric acid-glycine, the by-product of 21-esters could not be completely suppressed.

The present invention eliminates these drawbacks by allowing a hydrolytic enzyme active in the acidic region as described above to act on compound (I) under extremely mild conditions, thereby reducing the unfavorable amount in the process. .

17- of the target product without producing 21-esters.
This makes it possible to obtain only esters very easily, quantitatively, and in high yields, and eliminates the need for the step of removing 21-esters, which is essential in conventional methods.

For example, let us compare the results of the hydrolysis of dexamethasone 17α,21-methylorthotopionate according to the present invention with those obtained by conventional methods (using oxalic acid and using hydrochloric acid-sodium citrate buffer). .

The crude product was obtained by hydrolysis using a precoated thin silica gel plate containing a fluorescent reagent (manufactured by Merck & End Company) and a developing agent consisting of a hexane-chloroform acetone (4:3:3) mixture. The product was subjected to preparative thin layer chromatography in the usual manner, and the products were detected using a UV lamp. As a result, it was found that the standard product of dexamethasone 17α-propionate and the standard product of dexamethasone 21-gropionate were the same for both the conventional method and the painting method. Spots appeared at two locations with Rf values of 0.21 and 0.56, respectively, indicating that 17-esters and 21-esters were produced, but in the method of the present invention, dexamethasone 17α-propionate A spot appeared only at one location where the standard product had an Rf value of 0.21, clearly indicating that only 17-esters were produced.

In addition, in the method of the present invention, dexamethasone 17α,21-methylorthopropylene was hydrolyzed under the same conditions without the action of a hydrolytic enzyme, and then subjected to preparative thin layer chromatography in the same manner. A spot appeared only at one location where the standard pionate had an Rf value of 0.62.

This phenomenon occurs without adding an acid to the reaction solution and at a pH that is sufficient to absorb carbon dioxide from the atmosphere, 17α,21-
This shows that hydrolysis of orthoesters does not proceed at all.

However, in the present invention, even if the product has a hydroxy group other than the 17-position, these hydroxy groups are protected and the 17-position
There is no need for the complicated process of removing the protecting group again after esterifying the position.

In fact, not only do the various substituents and double bonds of the steroid ring not participate in the reaction of the present invention at all, but the 17α,21 cyclic orthoesters as starting materials are the corresponding 17α,21-dihydroxy The crude products produced from these compounds can be used as they are without purification.

Compound (I), which is a starting material used in the present invention, is composed of a corresponding 17α,21-dihydroxy compound having a partial structural formula and a general formula R-C(OR')3 (IV) (in which R
and R' have the same meanings as above.

) in an appropriate organic solvent such as dimethylformamide in the presence of an acidic catalyst such as toluenesulfonic acid or naphthalenesulfonic acid.
It can be easily produced by a known method such as heating at 0°C.

Orthoesters that can be used in this reaction include orthoacetic acid, orthopropionic acid, orthobutyric acid, orthoisobutyric acid, orthovaleric acid, orthoisovaleric acid, orthocaproic acid, orthoisocaproic acid, orthoenanthic acid, orthocaprylic acid, and orthocyclobutanecarboxylic acid. , orthocyclopentanecarboxylic acid, orthohexahydrobenzoic acid, orthobenzoic acid, and ortho-β-7enylpropionic acid. Among these, methyl ester or ethyl ester is preferable.

17α,2 produced using these orthoesters
All 1-cyclic orthoesters can be used as suitable starting materials for the present invention.

Therefore, the compound (I
) may have any type of steroid structure, but steroid structures having physiological activities such as anti-inflammatory effects, for example, the general formula (wherein R and R' have the same meanings as above).

R" represents a hydrogen atom, a methyl group or a hydroxy group at the α- or β-position.

Y1 represents a ketone oxygen atom, hydroxy group or halogen atom, Y2 represents a hydrogen atom or a halogen atom, or Y1 and Y2 are bonded to each other to represent an epoxy group.

Y3 represents a hydrogen atom, a halogen atom or a methyl group.

w1 and w2 each represent a hydrogen atom or a methyl group, or combine with each other to form a cyclopropane or double bond with the carbon atoms at the 1st and 2nd positions.

) It is desirable to have a corticosteroid structure such as the following.

In the compound represented by the general formula (2), there may be a double bond between the 1st and 2nd positions, between the 6th and 7th positions, or between the 9th and 11th positions, and there may be a double bond between the 18-nor form or the 19- It may be a Nor body.

In carrying out the present invention, the steroid structure may be a ring-nor structure, a ring-homocyclic structure, or an enol structure of each ketone group.

Compound II) obtained in the present invention is an adrenocortical hormone agent that is highly effective even as it is, and is effective in the treatment of rheumatism and allergy.

In particular, it not only has remarkable effects on various skin diseases as a topical antihypertensive agent, but also can be used as an intermediate for more important pharmaceuticals.

Next, the present invention will be explained with reference to Examples.

Example 1 Hydrocortisone 17-acetate [17α-acetoxypregnor-4-en-11β,21-diol-2
, 20-dione].

Hydrocortisone 17,21-methylorthoacetate [17α,21-(1-methoxy-1′-methyl-methylenedioxy)-pregnar-4-en-11β-ol-3,20-dione) (m., p. 223 ~225℃)
100 mg was dissolved in 35,771 liters of ethyl alcohol, to which was added a solution of pepsin 5I in 65 ml of purified water, and the mixture was shaken at 37° C. for 18 hours to react.

Ethyl alcohol was distilled off from the reaction solution under reduced pressure, and the mixture was extracted with chloroform.

The extract was washed with water, dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The resulting crude product was recrystallized from ethyl acetate to obtain the desired product.

Yield 95mg, m. p, 2 3 4-237℃
.

Example 2 Hydrocortisone 17-propionate [17α-propionyloxypreg-4-en-11β,21
-diol-3,20-dione].

(a) Hydrocortisone 17.21-Ethyl orthopropionate [17α,21-(1'ethoxy-1
7-ethyl-methylenedioxy)-Fregner-4-en-11β-ol-3,20-dione) (m.p.
18 2.5 to 18 3.5°C) 1001I1g
was treated in the same manner as in Example 1, and the obtained crude product was recrystallized from an acetone-hexane mixture to obtain the desired product.

Yield 96mg, m. p. 188-191°C.

(b) The raw material was treated in the same manner as in (a) except that 50 μg of the raw material was used and 2.5 g of bernase was used instead of pepsin.

Yield 44■.

(C) The same treatment as in (b) above was carried out using Takadiastase instead of Bernase.

Yield 44■.

Example 3 Preparation of hydrocortisone 17-7' thyrate [17α-butyryloxypregn-4-ene-11β21-diol 3,20-dione].

(a) Hydrocortisone 17.21-Mef-orthobutyrate [17α,21-(1'-methoxy1'
-propyl-methylenedioxy)-pregnar-4-ene-11β-ol-3,20-dione)
185.5 to 187.5°C) at 100° C., and treated in the same manner as in Example 1, and the obtained crude product was recrystallized from a mixture of ethyl acetate and hexane to obtain the desired product. .

Yield 9 4 [Qg, m, p. 205-207
℃.

(b) The same treatment as in (a) above was carried out using vanprosin instead of pepsin.

Yield 91tllg0 Example 4 Hydrocortisone 17-valerate [17αy<L
/ IJ Lux-pregnath-4-ene-11β,21
-diol-3,20-dione].

Humanocortisone 17.21-Methylorthovalerate [17α,21-(1'-butyl-1'-methoxymethylenedioxy)-pregnar-4-en-11β-ol-3,20-dione] (m.p. 200 ~204°
C) is 100tIlg, 5g morsin instead of pepsin
The crude product obtained was recrystallized from an acetone-hexane mixture to obtain the desired product.

Yield 9 4tllg, m. p, 1 5 8 ~ 1
6 2°co Example 5 Preparation of hydrocortisone 17-caproate (17α-cagloyloxypregn-4-ene-11β,21-diol-3,20-dione).

Hydrocortisone 17.21-Methyl orthocaproate [17α,21-(1'-methoxy1'=pentyl-methylenedioxy)-flexner-4-en-11β
-ol-3,20-dione] (m.p. 119-12
0°C) 50■ dissolved in 20ml of ethyl alcohol,
A solution of 2.5 g of vanprosin dissolved in 30 d of purified water was added thereto, and the mixture was stirred at room temperature for 3 days to react, and the reaction solution was treated in the same manner as in Example 1.

The obtained crude product was separated by preparative thin layer chromatography using a hexane-chloroform acetone (4:3:3) mixture as a developing agent, and recrystallized from an acetone-hexane mixture to obtain the desired product. Ta.

Yield: 35 ■, m, p, 156-159°C.

Example 6 Hydrocortisone 17-cyclopentanecarboxylate [17α-cyclopentanecarbonyloxypregn-4-ene-11β,21-diol-3,20-
Zeon] production.

Hydrocortisone 17.21-Methylorthosicpentanecarboxylate [17α,21-(1'-cyclopentyl-1'-methoxymethylenedioxy)-phlegnar-4-ene-11β,21-diol-3,20
-Zion Em. p. 197~201°C) to 100[1
]g and Bernase 5g in the same manner as in Example 1.

The obtained crude product was recrystallized from an acetone-hexane mixture to obtain the desired product.

Yield 9 2tQg, m. p, 2 3 0~2 3
2℃.

Example 7 Hydrocortisone 17-penzoate [17α-penzoyloxypreg f-4-y-11β,21-
Production of diol-3,20-dione].

(a) Hydrocortisone 17.21-methylorthobenzoate [17α,21-(1'-methoxy-1
'-phenyl-methylenedioxy)-pregnar-4-ene-11β-ol-3,20-dione) (m.p.2
using 100 μg of os~210°C) and 5 g of pepsin.
It was treated in the same manner as in Example 1.

The obtained crude product was recrystallized from an acetone-hexane mixture to obtain the desired product.

Yield: 90tQg, m. p, 2 2 0~2
2 2℃.

(b) The same treatment as in (a) above was carried out using bernase instead of pepsin.

Yield: 88 [Ilgo (C) 50 mg of the raw material was treated in the same manner as in (a) above, using 10 p of denapsin instead of pepsin.

Yield: 40tIlg0 (d) In (a) above, the raw material was 20μ, lipase APl& was used instead of pepsin, and the reaction time was 36 hours, and the same treatment as in Example 1 was performed.

The obtained crude product was separated by preparative thin layer chromatography using a hexane-chloroform acetone (4:3:3) mixture as a developing agent, and recrystallized from an acetone-hexane mixture to obtain the desired product. Ta.

Yield 17■.

Example 8 Preparation of prednisolone 17-acetate [17α-acetoxypregner 1,4-diene-11β,21-diol-3,20-dione].

Prednisolone 17,21-methylorthoacetate [17α,21-(1'-methoxy-1'-methyl-methylenedioxy)-pregner 1,4-diene-11β
-ol-3,20-dione] (m.p. 185-18
The treatment was carried out in the same manner as in Example 1 using 100 μm of 8°C.

The obtained crude product was recrystallized from a mixture of aceto-hexane to obtain the desired product.

Yield: 92cm. p. 240-242℃.

Example 9 Prednisolone 17-propionate [17α-propionyloxy-pregna-1,4-diene-11β,
21-diol-3,20-dione].

Prednisolone 17.21-Ethyl orthofuropionate [17α,21-(1'-ethoxy-1'-ethyl-methylenedioxy)-pregner 1,4-diene-1
1β-ol-3,20-dione] (m.p. 180~
The treatment was carried out in the same manner as in Example 1 using 1 g of 1001I (184°C).

The obtained crude product was recrystallized from acetone-11/xane mixture to obtain the desired product.

Yield: 96cm. p. 207-212℃.

Example 10 Production of prednisolone 17-butyre} (17α-butyryloxy-pregna-1,4-diene-11β,21-diol-3,20-dione).

Prednisolone 17.21-Methyl orthobutyrate [17α,21-(1'-methoxy-1'-propyl-methylenedioxy)-pregner 1,4-diene-11
β-ol-3,20-dione] (m.p, 1 6
5-168°C) 100mg, Bernase 5g
The same treatment as in Example 1 was carried out using the same method.

The obtained crude product was recrystallized from an acetone-hexane mixture to obtain the desired product.

Yield 92mg, m. p. 203-207°co Example 11 Prednisolone Preparation of 17-valerate [17α-valeryloxypregna-1,4-diene-11β,21-diol-3,20-dione].

Prednisolone 17.21-Methylorthovalerate [17α,21-(1'-butyl-1'-methoxymethylenedioxy)-pregner 1,4-diene-11β
-ol-3,20-dione] (m, p, 1 5
The treatment was carried out in the same manner as in Example 1 using 100 mg of 7-159°C) and 5 g of vanprosin.

The obtained crude product was recrystallized from an acetone-hexane mixture to obtain the desired product.

Yield 9 6[I1g, m, p, 2 1 0~2
1 3℃.

Example 12 Betamethasone 17-acetate [17α-acetoxy 9α-fluoro-16β-methylpregner 1,4
-Diene-11β,21-diol-3,20-dione].

(a) Betamethasone 17,21-ethyl orthoacetate [9α-fluoro-16β-methyl-17α
,21-(1-ethoxy-1'-methyl-methylenedioxy)-pregner 1,4-diene-11β-ol-
Example 1 Using 1001I1g of 3,20-dione] (m.p. 156-157°C) and 5g of pepsin.
processed in the same way.

The obtained crude product was recrystallized from an acetone-hexane mixture to obtain the desired product.

Yield 9 4fQgXm, p. 2 4 0～2 4
2°C (decomposition).

(b) The desired product was obtained by treating in the same manner as in (a) above using linivernase instead of pepsin.

Yield: 95111go (C) In (a) above, treat in the same manner as in Example 6(d) using pepsin substitute linicellulase AP, and recrystallize the obtained crude product from an acetone-hexane mixture to obtain the target product. I got it.

Yield 93 [I! go Example 13 Betamethasone 17-propionate [9α-fluoro-16β-methyl-17α-propionyloxy-pregnathic-1,4-diene-11β,21-diol-3
, 20-dione].

(a) Betamethasone 17.21-Ethyl orthopropionate [9α-fluoro-16β-methyl-1
7α,21-(1'-ethoxy-1'-ethyl-methylenedioxy)-pregner 1,4-diene-11β-ol-3,20-dione: (m.p. 208-211°
C) Treated in the same manner as in Example 2 using 100 tllg and 5 g of pepsin.

The obtained crude product was recrystallized from acetone to obtain the desired product.

Yield 95 [ng, m. p. 231~235°co(b
) The desired product was obtained in the same manner as in (a) above using linivernase instead of pepsin.

Yield: 95Ing0 (C) In the above (a), biodiastase was used instead of pepsin and the target product was obtained by the same treatment. Yield: 92[ng0 (d) In the above (a), when Sunprose F was used instead of pepsin The desired product was obtained by the same treatment.

Yield: 92tIlg0 (e) The desired product was obtained in the same manner as in (a) above using panprosin instead of pepsin.

Yield: 96 [og0 0) The same procedure as in (a) above was carried out except that 50 mg of the raw material was used, 2.5 g of vancerase was used instead of pepsin, and the reaction time was 36 hours.

The obtained crude product was separated by preparative thin layer chromatography in the same manner as in Example 5, and recrystallized from acetone to obtain the desired product.

Yield 45 mgo Example 14 Betamethasone 17-butyrate [17α-butyryloxy-9α-fluoro 16β-methyl-pregner 1,4-diene-11β,21-diol-3,20-
Zeon] production.

Gloss Betamethasone 17.21-Methyl orthobutyrate [9α-fluoro-16β-methyl-17α,21
-(1'-methoxy1'-propyl-methylenedioxy)-pregner 1,4-diene-11β-ol-3
, 20-dione] (m.p. 148°C) in the same manner as in Example 1, and the resulting crude product was recrystallized from an acetone-hexane mixture to obtain the desired product.

Yield 9 6+1tgom. p, 193~19
5℃.

A) The desired product was obtained by treating in the same manner as in (a) above using linivernase instead of pepsin.

(C) The same treatment as in (C) above was carried out using Takadiastase instead of pepsin, and the resulting crude product was recrystallized from acetone to obtain the desired product.

Yield 9 4[I1g0 Example 15 Betamethasone 17-inbutyrate [9α-fluoro-17α-isobutyryloxy-16β-methyl-pregner 1,4-diene-11β,21-diol-3
, 20-dione].

(P) Betamethasone 17,21-methylorthoisoptylate [9α-7 luorol 16β ~ methyl-1
7α,21-(1'-isoprobyl-1'-methoxymethylenedioxy)-pregner 1,4-diene-11
β-ol-3,20-dione) (m, p, 1 7
3°C) 100[I1g and Vanprosyn 5I were used in the same manner as in Example 1, and the resulting crude product was recrystallized from an acetone-ether mixture to obtain the desired product.

Yield 95mg, m. p. 245-248°C (decomposition)
.

(b) The target product was obtained in the same manner as in (a) above using linivernase instead of vanprosin.

Yield 90mg0 Example 16 Betamethasone 17-valerate [9α-F#:)r
O-1 6β~methyl-17α-valeryloxypregna-1,4-diene-11β,21-diol-3
, 20-dione].

(a) Betamethasone 17.21-Methyl orthovalerate [9α-fluoro-16β-methyl-17α,2
1-(1'-butyl-1'-methoxymethylenedioxy)-pregner 1,4-diene-11β-ol-3
,20-dione] (m.p. 152-154°C) 10
The crude product obtained was recrystallized from an acetone-hexane mixture to obtain the desired product.

Yield 9 6mg, m, p. 1 8 2 ~1
8 4°co (b) The desired product was obtained by treating in the same manner as in (a) above, using bernase instead of pepsin.

Yield: 97 [I1g0 (C) The desired product was obtained by performing the same treatment as in (a) above using morsin instead of pepsin.

Yield 95tng.

(d) The desired product was obtained in the same manner as in (a) above using linibanprosin instead of pepsin.

Yield 97I11go Example 17 Betamethasone 17-isovalerate [9α-fluoro-17α-invaleryloxy-16β-methyl-pregner 1,4-diene-11β,21-diol-3
, 20-dione].

Betamethasone 17.21-Methyl orthoisovalerate [9α-fluoro16β-methyl = 17α,21
-(1'-isobutyl-1'-methoxymethylenedioxy)-pregner 1,4-diene-11β-ol-
3,20-dione] (mp, 175-176°C)
The resulting crude product was treated in the same manner as in Example 1 using 100 mg and Bernase 5I, and the resulting crude product was recrystallized from an acetone-hexane mixture to obtain the desired product.

Yield 95mg, m, p. 2 1 9-221℃
.

Example 18 Betamethasone 17-cabroate [9α-fluoro-17α-caproyloxy-16β-methyl-pregner 1,4-diene-11β,21-diol-3,2
0-dione] production.

Betamethasone 17.21-Methyl orthocaproate [9α-fluoro16β-methyl-17α,21-
(1'-methoxy1'-pentyl-methylenedioxy)-pregnane-1,4-diene-11β-ol-3
, 20-dione) (m.p. 148-151°C) 100
The crude product thus obtained was recrystallized from an acetone-hexane mixture to obtain the desired product.

Yield 9 4 [Qg, m. p, 1 4 8-150
°co Example 19 Betamethasone 17-cyclopentanecarbo-11'
-Sylate [17α-cyclopentanecarbonyloxy-9α-fluoro-16β-methyl-pregner 1,4
-Diene-11β,21-diol-3,20-dione].

Betamethasone 17.21-Methylorthocyclopentanecarboxylate [9α-7luol-16β-methyl-17α,21-(1'-cyclopentyl-17-methoxymethylenedioxy)-pregner 1,4-diene-11β-ol-3 ,20-dione]50[I1g
The crude product obtained was treated in the same manner as in Example 1 using pepsin 2.59 and recrystallized from an acetone-hexane mixture to obtain the desired product.

Yield 4 4mg, m. p. 2 2 7-230℃.

Example 20 betamethasone 17-benzoe} (17α-pensoyloxy-9α-fluoro 16β-methyl-pregner 1,4-diene-11β,21-diol-3,2
0-dione] production.

(a) Betamethasone 17.21-Methyl orthobenzoate [9α-fluoro-16β-methyl-17
α,21-(1'-methoxy-1 phenyl-methylenedioxy)-pregner 1,4-diene-11β-ol-3,20-dione] (m.p. 169-172°G
) and 2.5 g of pepsin were treated in the same manner as in Example 1, and the resulting crude product was recrystallized from an acetone-ether mixture to obtain the desired product.

Yield 45 [ng, m. p. 227~230°co(b
) The desired product was obtained in the same manner as in (a) above, using bernase instead of pepsin.

Yield 4 3mg, m. p, 2 2 5 ~ 2 3
0°co (C) The same treatment as in (a) above was performed except that lipase AP was used instead of vepsin and the reaction time was 40 hours.

The obtained crude product was separated by preparative thin layer chromatography in the same manner as in Example 5, and recrystallized from acetone-ether to obtain the desired product.

Yield 38 [IIg, m. p. 224-229°C.

Example 21 Preparation of 9α-chloro-16β-methyl-17α-fropionyloxypregna-1,4-diene-11β,21-diol-3,20-dione.

9α-chloro16β-methyl-17α,21-(1′
monoethyl-17-methoxymethylenedioxy)-pregna-1,4-diene-11β-ol-3,20-dione (m, p. 174-178'C) 100tllg,
The treatment was carried out in the same manner as in Example 1, using 5 g of bernase and a reaction time of 24 hours.

The obtained crude product was recrystallized from an acetone-hexane mixture to obtain the desired product.

Yield: 96cm. p, 197-199°C (decomposition) C Example 22 Preparation of 17α-acetoxy9β,11β-epoxy pregna-4-en-11β-ol-3,20-dione.

9β,11β-epoxy-17α,21-(1'-methoxy-1'-methyl-methylenedioxy)-pregnar-4-ene-3,20-dione (m.p. 157-161
The treatment was carried out in the same manner as in Example 1, using 50 [ng (°C) and 2.5 g of bernase, and the reaction time was 24 hours.

The obtained crude product was recrystallized from an acetone-ether mixture to obtain the desired product.

Yield: 46mg, m. p. 1 9 3-195°C.

Example 23 Preparation of 17α-penzoyloxy-9β,11β-epoxy 16β-methyl-pregner 1,4-dien-21-ol-3,20-dione.

9β,11β-epoxy-16β-methyl-17α,2
Using 100 mg of 1-(1'-methoxy-1'-7enyl-methylenedioxy)-pregner 1,4-diene-3,20-dione (m.p. 148-150°C) and 5 g of pepsin. , and treated in the same manner as in Example 1.

The obtained crude product was recrystallized from an acetone-hexane mixture to obtain the desired product.

Yield: 94 Ing, m. p. 135-138°C.

Example 24 Dexamethasone 17-acetate [17α-acetoxy 9α-fluoro 16α-methyl-pregner 1,
4-diene-11β,21-diol-3,20-dione].

Dexamethasone 17.21-Methyl orthoacetate [9α-fluoro-16α-methyl-17α,21-(
1'-methoxy-1'-methyl-methylenedioxy) monopregner 1,4-diene-11β-ol-3,20
-dione] (m.p. 200-202°C) 5 0 [
1]g and Bernase 2.5g, the reaction time was 22 hours, and the same treatment as in Example 1 was carried out.

The obtained crude product was recrystallized from an acetone-hexane mixture to obtain the desired product.

Yield: 97 [I] g, m. p. 206-211℃.

Example 25 Dexamethasone 17-propionate [9α-fluoro-16α-methyl-17α-propionyloxypregna-1,4-diene-11β,21-diol-3
, 20-dione].

(a) Dexamethasone 17.21-Ethyl orthopropionate [9α-fluoro-16α-methyl-1
7α,21-(1'-ethoxy-1'-ethyl-methylenedioxy)-Gregner 1,4-diene-11β-ol-3,20-dione] (m.p. 219-221°
C) Treated in the same manner as in Example 1 using 100 Ing.

The obtained crude product was recrystallized from an acetone-hexane mixture to obtain the desired product.

Yield 9'7111gy m. l). 2 1 8～2
21°C (b) The desired product was obtained by treating in the same manner as in (a) above, using bernase instead of pepsin.

Yield: 95tng0 (C) The desired product was obtained by performing the same treatment as in (a) above using Takadiastase instead of pepsin.

Yield 96 [I1go Example 26 Dexamethasone 17-butyrate [17α-butyryloxy-9α-fluoro 16α-methyl-pregner 1,4-diene-11β,21-diol-3,20-
Zeon] production.

(a) Dexamethasone 17.21-Methyl orthobutyrate [9α-fluoro-16α-methyl-17α
,21-(1'-methoxy1'-propyl-methylenedioxy)-pregner 1,4-diene-11β-ol-3,20-dione] (m.p, 180-1
86° C.) and 2.5 g of pepsin.

The obtained crude product was recrystallized from an acetone-petroleum ether mixture to obtain the desired product.

Yield 4 61Qg, m, p, 190~19
3℃.

(b) The target product was obtained by treating in the same manner as in (a) above using linivernase, which is a substitute for pepsin.

Yield 45mg0 Example 27 Dexamethasone 17-valerate [9α-7luoro-16α-methyl-17α~valeryloxypregna-
1,4-diene-11β,21-diol-3,20-
Zeon] production.

Dexamethasone 17.21-Methyl orthovalerate [9α-fluoro-16α-methyl-17α,21-(
1'-butyl-1'-methoxymethylenedioxy)-Flegner 1,4-diene-11β-ol-3,20
-dione) (m.p. 184-185°C) 10011
The same treatment as in Example 1 was carried out using 1 g.

The obtained crude product was recrystallized from an acetone-hexane mixture to obtain the desired product.

Yield 96mg, m. p, 17 9-181℃
.

Example 28 Dexamethasone 17-inbutyrate [9α-monofluoro 17α-isobutyryloxy-16α-methyl-pregner 1,4-diene-11β,21-diol-3
, 20-dione].

Dexamethasone 17.21-Methyl orthoisoptylate [9α-fluoro-16α-methyl 17α,21-
(1'-isoprobyl-1'-methoxymethylenedioxy)-pregner 1,4-diene-11β-ol-
3,20-dione] (m.p. 212-219°C) 1
The same treatment as in Example 1 was carried out using 00tllg.

The obtained crude product was recrystallized from an acetone-hexane mixture to obtain the desired product.

Claims (1)

[Scope of Claims] 1 17α,21-dihydroxy-20-ketopregnan having a partial structural formula (wherein R represents an alkyl group, a cycloalkyl group, or an aryl group, and R' represents a lower alkyl group) 21- having a partial structural formula (wherein R has the same meaning as above) characterized by allowing a hydrolase having activity in an acidic region to act on a 17α,21 cyclic orthoester of a series steroid. A method for producing hydroxy-20-ketopregnan steroid 17-esters.