JPS5846505B2 - Method for producing 1χ,24-dihydroxycholecalciferol - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing 1α,24-dihydroxycholecalciferol represented by the following formula (3).

Conventionally, the compound has been produced by deriving it from naturally occurring fucosterol, but although fucosterol is useful as a raw material, its production relies entirely on extraction from plants; and
It is extremely difficult to obtain as a raw material and is not preferred for industrial production.

According to the present invention, since it is a readily available raw material, 1
This made it possible to use tocoric acid, which could not be used in the synthesis of α,24-dihydroxycholecalciferol.

That is, the present invention provides 3,24-dihydroxycolescune represented by formula (I) in a solvent with 2,3-dichloro-5,6
-dicyazbenzoquinone to give 24-hydroxy-cholest-1,4-dien-3-one of formula (n), followed by 2,3-cyclo5,6-
Treated with loraniil, which contains dithiazhydroquinone,
The produced 24-hydroxy-cholest-1,4,64-dryen-3-one@) is isomerized in the presence of a basic catalyst to form 24-hydroxy-cholest-1,5,7-dryen-3-one, and this is reduced with metal hydride to give 3β, 24
-dihydroxy-cholesta-1,5,7-triene (7
), and then add 1°2.4 represented by the general formula (to) (in the formula, R means an organic residue).
1 represented by the general formula (7) (wherein R means the same as above),
4-El adduct, reacted with peroxide,
1α represented by the general formula (g) (in the formula, R means the same as above)
, 2α-epoxide compound, and then reduced with metal hydride to produce 1α and 3β.

1α,24-, which is characterized by irradiating 24-trihydroxy-cholesta-5,7-diene (b) with ultraviolet rays to form 1α,24-dihydroxybrevicun D3(1)O, and then isomerizing this. This invention relates to a method for producing dihydroxycholecalciferol ω.

Note that 3,24-dihydroxycholestane (I), which is the starting material of the present invention, is a new compound and is easily produced using lithocholic acid as a raw material.

An example of the manufacturing method is shown below.

When producing (If) from compound (I) in the present invention, the reaction is carried out in a solvent. Examples of solvents that can be used include ethers such as dioxane, and n-amyl alcohol.

Higher alcohols such as 5eC-amyl alcohol and hydrocarbon solvents such as xylene can be used, but dioxane and n-amyl alcohol are preferred.

The reaction temperature is preferably the reflux temperature of the solvent used (usually 100°C or higher).

The reaction time is usually appropriately selected within the range of 10 to 50 hours, but the reaction time can be further shortened by lowering the ratio of solvent to compound (1).

Further, the amount of 2,3-dichloro-5,6-dicyazbenzoquinone used is required to be 3 molar equivalents or more based on compound (1).

Compound (n) can be isolated from the reaction mixture by a conventional method, for example, after cooling the reaction mixture, the precipitated crystals (2,3-dichloro-5,6-diciatubideroquinone) are removed in an oven, and the filtrate is concentrated. The reaction mixture is then subjected to a means such as column chromatography, but the reaction mixture can also be subjected to the next step without being particularly purified.

Compound (4) is obtained by treating compound (II) with floranil in the presence of 2,3-dichloro-5,6-diciatubideroquinone.

At this time, the reaction is preferably carried out in a solvent, and examples of solvents that can be used include ethers such as dioxane and higher alcohols such as n-amyl alcohol and 5eC-amyl alcohol.

The reaction temperature is preferably the reflux temperature of the solvent used.

The reaction time is usually several tens of hours, but when compound (II)
The reaction time can also be shortened by lowering the ratio of solvent to

The additive 2,3-dichloro-5,6-diciatubideroquinone is required in an amount of 1 molar equivalent or more, and the amount of Floranil is required in an amount of 1 molar equivalent or more relative to the compound (If).

Compound (4) is isolated from the reaction mixture by a conventional method, for example, by distilling off the solvent and subjecting the residue to a means such as column chromatography.

In the step of isomerizing compound (2) with a basic substance to produce compound (5), any ordinary basic substance can be used as the basic substance, but preferably sodium methoxide, sodium ethoxide, potassium-t-
Alcoholades such as butoxide.

The reaction is preferably carried out in a solvent, and the solvent is preferably one in which compound (III) has excellent solubility;
Ethers, benzene solvents, hydrocarbon solvents and alcohols are used.

Note that it may be advantageous to use DMSO in relation to the basic substance used, and DMSO and potassium-t
- The combination of butoxide gives excellent results in terms of reactivity.

The reaction temperature is appropriately selected from cold to heated.

Note that this isomerization reaction is preferably carried out in an inert gas atmosphere such as nitrogen or argon.

Compound (5) is isolated from the reaction mixture by a conventional method, for example, by neutralizing the reaction solution, extracting it, concentrating the extract, and subjecting it to a means such as column chromatography. From the viewpoint of yield, it is advantageous to proceed to the next step without removing the raw material.

Compound M is obtained by reducing the compound (5) thus obtained with a metal hydride.

Examples of metal hydrides used in this case include lithium aluminum hydride and sodium borohydride.

Calcium borohydride, etc.

The reaction is preferably carried out in a solvent, and the most suitable solvent is an ether solvent, such as ether, tetrahydrofuran, 1,2-dimethoxyethane, diglyme, etc.

In addition, even if the metal hydride is sodium borohydride,
When using borohydrides such as calcium borohydride, a solvent having a hydroxyl group such as water, methanol, and ethanol can also be used.

The reaction temperature is appropriately selected from cold to heated.

Isolation of (7) from the reaction mixture is carried out by a conventional method, for example, by neutralizing the reaction solution, extracting it, concentrating the extract, and subjecting it to a means such as column chromatography. It is also possible to proceed to the next step without removing it.

Compound (b) is produced by reacting compound (b) with compound (b) thus obtained.

At this time, the reaction is preferably carried out in a solvent.

As the solvent, any solvent can be used as long as it is inert during the reaction, but organic solvents such as ethers, benzene-based solvents, and hydrocarbon-based solvents are generally preferred, and specifically benzene.

These include tetrahydrofuran and methylene chloride.

The reaction temperature is appropriately selected between cold and heated to reflux.

General formula (1,2,4-) IJ azoline-3
In the 5-dione derivative, R can be any organic residue that is inactive during the reaction, but it is usually a monocyclic aromatic residue because of its excellent solubility in the solvent used.
A lower alkyl group is used, specifically a phenyl group, a monosubstituted phenyl group, a methyl group, an ethyl group, etc.

Isolation of the compound (material) from the reaction mixture is carried out by concentrating the reaction solution, extracting it with a solvent such as benzene or methylene chloride, and then subjecting it to column chromatography.

The compound (VD) thus obtained is reacted with a peroxide to produce compound (1).

The peroxide is preferably an organic peracid.

As the organic peracid, aromatic peracids such as perbenzoic acid (2m-chloroperbenzoic acid) and aliphatic peracids such as permaleic acid, peracetic acid, and pertrifluoroacetic acid can be used.

The reaction is preferably carried out in a solvent, and examples of solvents that can be used include ethers, ethers such as tetrahydrofuran, hydrocarbon solvents such as chloroform and methylene chloride, alkyl esters of organic acids such as ethyl acetate, There are fatty acids such as acetic acid and acetic acid, but hydrocarbon solvents such as methylene chloride and chloroform are most suitable because they are extremely inert to peroxides and have excellent solubility for the compounds used.

Note that salts such as sodium carbonate and disodium hydrogen phosphate can also be used as a buffer in order to advance the reaction advantageously.

The reaction temperature is appropriately selected between cold and heated depending on the type of peroxide used.

(4) is obtained by reducing (a) with a metal hydride.

The hydride metal used in this case is preferably lithium aluminum hydride, lithium borohydride, or the like.

Preferably, the reaction is carried out in a solvent.

Ether solvents are most suitable as solvents, specifically ether and tetrahydrofuran.

1,2-dimethoxyethane, diglyme, etc.

The reaction temperature is appropriately selected between room temperature and reflux temperature.

Isolation of (1) from the reaction mixture is carried out by a conventional method, for example, by decomposing the excess metal hydride, extracting it, concentrating the extract, and subjecting it to a means such as column chromatography.

By irradiating (2) with ultraviolet rays, the B ring of the steroid skeleton of (3) was cleaved (Do was produced).

It is appropriate to carry out this ultraviolet irradiation in a solvent, and examples of solvents that can be used include hydrocarbon solvents, ether solvents, and alcohols.

Examples of the hydrocarbon solvent include saturated hydrocarbon solvents, among which those with a low boiling point are preferred, and specific examples include hexane, octane, and the like.

The ether solvent is preferably a saturated one, most preferably a low boiling point saturated ether, specifically ether, tetrahydrofuran, etc.

Examples of alcohols include lower alcohols, specifically methanol.

The reaction is suitably carried out at a temperature around room temperature, and preferably in an atmosphere of an inert gas such as argon.

The irradiation time of ultraviolet rays varies depending on the intensity of the light source lamp and the scale of the reaction, and is appropriately selected from several tens of seconds to several hours.

Isolation of (7) from the reaction mixture is carried out by a conventional method, for example, after concentration, by means such as chromatography.
It can also be directly subjected to the next step without isolation.

Compound (1) and target compound (2) are in a thermal equilibrium state, and (1) is isomerized to produce (2).

This isomerization reaction is carried out in a conventional manner by standing in a solvent in a dark place or by heating in a solvent.

The solvent at this time may be any solvent that is inert during the reaction, and common hydrocarbon solvents, ether solvents, alcohols, and benzene solvents are used.

Preferably, it has a low boiling point and excellent solubility of compound (1), and specific examples include hexane, isooctane, toluene, ether, and tetrahydrofuran.

The reaction time is appropriately selected from several hours to several weeks depending on the reaction conditions.

When left in a solvent at room temperature, several weeks is appropriate, and when heated under reflux, several hours is appropriate.

Note that this isomerization reaction is preferably carried out in an atmosphere of an inert gas such as nitrogen or argon.

The target compound (1) is isolated from the reaction mixture by a conventional method.
For example, it is purified by extraction, recrystallization, column chromatography, partition chromatography, etc., especially Sephadex (product of Pharmacia Fine Chemicals).
Column chromatography packed with

Although compounds (goods) that are not isomerized in this isomerization step are also obtained, after recovery, (1) can be further subjected to this isomerization reaction.

In the present invention, compounds (I), (It), Q
V), (V), (VDO) and (V) are all new compounds that have not been described in any literature.

Hereinafter, embodiments of the present invention will be specifically explained with reference to Examples.

Example a) Dry magnesium turnings 7, 27 f in ether 2
00rnl and isopropyl bromide under stirring at room temperature.
．． A solution of 1r111 dissolved in dry ether 1oo- is added dropwise, and after the dropwise addition it is boiled for 30 minutes to completely dissolve the magnesium.

After cooling, 43.56 ff of anhydrous cadmium bromide powder is gradually added.

After boiling for 1 hour, ether was distilled off, and dry benzene was added to replace the solvent (until the boiling point reached 80°C).

On the other hand, acetyl lithocholic acid 102 and thionyl chloride 100
A solution was prepared by dissolving acetyl lithocholic acid chloride prepared by dissolving it in dry benzene by dissolving excess thionyl chloride and leaving it at room temperature for 2 hours, and added it to the benzene solution of isopropyl cadmium. Add dropwise at 5-7° C. with vigorous stirring.

After 5 to 10 minutes of dropping, cool and add 5-hydrochloric acid to decompose the reagent, then add new benzene, wash the benzene layer with water, dry with magnesium sulfate, and distill off the solvent to a size of about 300 m.

To this, a methanol solution of 1N potassium hydroxide 100-
was added, left overnight at room temperature, and extracted with ether.

After washing with water and drying the upper layer, the solvent is distilled off to precipitate crystals.

After thorough washing with petroleum ether, 4.66f (48,4φ) of 5β-cholestan-24-one-3α-ol is obtained.

Melting point 123.5-125℃ Mass spectrum: m/e 402 (M+).

394.379 b) Dissolve 5β-cholestan-24-one-3α-ol 187.31119 in methanol, add a solution of sodium borohydride 90'/V in methanol under water cooling, stir for 1 hour, and acidify with hydrochloric acid. It is then extracted with ether, washed with water, washed with sodium bicarbonate solution, further washed with water, dried over magnesium sulfate, and the solvent is evaporated.

185 m9 (98,3 pieces) of needle-like crystals of 5β-cholestane-3α,24-diol were obtained from methanol.

Elemental analysis value: CH as C27H4802 Theoretical value @ 80.14 11.96 Actual value :chloroform), the 24-epimer can be easily separated.

5β-cholestane-3α,24ε1-diol White needle crystals (tess polar) Melting point 123-124.5°C
5β-Cholestane-3α,24ε2-diol White needle crystals (more polar) Melting point 143.5-144.
5 DEG C. c) 117 ml of 5β-cholestane-24,3α-diol was dissolved in 2 rnl of dry dioxane, 217 ml of 2,3 dichloro-5,6-diciatzbenzoquinone was added, and the mixture was heated under reflux for 20 hours.

The following treatment is carried out in a conventional manner to obtain oily 24-hydroxy-cholester-1,4-dien-3-one (44cm) (38.2cm).
get.

UV spectrum: λ”10H243nm NMR spectrum: δ (throat) 3.1-3.5 (IH, m
) 6.09 (IH, d) 6.19 (IH.

dd) 7.07 (IH, d) Mass spectrum: m/e 398 (M”).

80 d) 24-hydroxy-cholesta-1,4-diene-3
400 mg of -one was dissolved in 35 cm of dioxane, 300 μl of Floranil and 150 μl of 2,3-dichloro-5,6-diciatubideroquinone were added, and the mixture was heated under reflux for 40 hours.

The following treatment is carried out in a conventional manner to obtain 24-hydroxy-cholester.
1,4,6-dorien-3-on 24011! get.

UV spectrum: λ”0H299,257.

Geki π 243 nm NMR spectrum (CDC4): δ (pP) 3.1-3
．． 5 (LH, m), 5.95-6.2 (3H,
m), 6.29 (IH, dd).

7.11 (IH, d) Mass spectrum: m/e 396 (M+).

78 e) Add 100 - of dry dimethyl sulfoxide to the reaction vessel and then dissolve 3.62 - of 24-hydroxycholesta 1.4,64 lien-3-one.

Fill the container with dry argon gas.

While stirring the solution vigorously at room temperature, potassium 22
and powdered potassium t-butoxide prepared from 80 ml of t-butanol were added all at once.

The reaction solution is vigorously stirred for an additional 30 minutes and then poured into ice water saturated with carbon dioxide gas by the addition of dry ice.

Next, extract with ether pre-cooled to 0 to -5°C several times until the aqueous layer becomes clear.

The extract was quickly washed with a large amount of cooling water saturated with carbon dioxide gas until the washing liquid became neutral, and the ether was distilled off, resulting in a crude 2
4-Hydroxycholester-1,5,7-t-liene-3
- Get about 32 on.

f) Sodium borohydride 3.61 parts ethanol 100 parts
- Calcium chloride 7 in which the solution is cooled to below -10°C
, Ov into a solution of 150 ml of methanol with stirring.

After continuing stirring for 20 minutes while maintaining the temperature at -10℃,
24-hydroxy-cholester-1 obtained in e) above,
5゜7-) Ether 100 containing IJ en-3-one
- solution under stirring.

The reaction solution was further stirred at -10°C for 1 hour, and then diluted with 10%
Neutralize with acetic acid, add 200ml of water, and extract with ether.

The ether layer was washed with water, dried with anhydrous magnesium sulfate,
When the ether is distilled off, crude 3β is obtained.

24-dihydroxycholester-1,5,7-triene (
Approximately 3f) is obtained.

g) 100 containing the crude 3β,24-dihydroxycholester-1,5,7-)riene (approximately 3r) obtained in f) above.
- to the ether solution of 4-phenyl-1,2,4-triazoline-3,5-dione is added little by little with stirring until the ether solution remains red.

After stirring for 1 hour at room temperature, the ether was distilled off and the residue was chromatographed on a column packed with silica gel 151.

The effluent fractions were collected with chloroform containing 1 cup of methanol, and 3β,24-dihydroxycholester-1,5,7
-) 1,4-cycloadduct 2.12 of liene and 4-phenyl-1,2°4-triazoline-3,5-dione is obtained.

Elemental analysis value: CHN calculated value (to) 73.278.267.32 as C35H4□04N3 Actual value (equity)
72.878.207.39h) 1 obtained in g) above
, 4-m adduct 1.3262 was dissolved in 50-chloroform, 1.51' of m-chloroperbenzoic acid was added thereto, and the mixture was stirred at room temperature for 48 hours.

The reaction solution was diluted with chloroform, washed with 10 polypotassium carbonate aqueous solution, dried over anhydrous magnesium sulfate, and the chloride agent was distilled off under reduced pressure.

The residue was subjected to silica gel column chromatography,
Collecting the first fraction flowing out with ether containing 1% methanol and recrystallizing it from ether gives a melting point of 162-16.
The 1β,2β-epoxide of the 1.4-JJ adduct was obtained at 4°C, and the subsequent fractions were collected and recrystallized from ether containing 10 methanol to give a melting point of 205-20.
1α,2α-epoxide of 1,4-cycloadduct at 8°C 7
17.5■ is obtained.

Elemental analysis value: CHN as C35H4705Na Calculated value (to) 71.288.037.12 Actual value (to)
70.827.857.091) 3 obtained in h) above
β,24-dihydroxycholester-1,5,7-triene and 4-phenyl-1,2,4-triazoline-3,5
89.2 mL of 1α, 2α-epoxide compound of 1,4-cycloadduct of -dione! was dissolved in anhydrous tetrahydrofuran (10), and lithium aluminum hydride (8) was added to the solution.
Add 9'IIIi little by little while stirring.

Then, the mixture was secretly boiled under reflux for 1 hour, and after cooling, a saturated aqueous solution of Glauber's salt was added until no more foaming occurred to decompose excess lithium aluminum hydride. The organic solvent layer was separated, dried, and the solvent was distilled off.

The residue is purified by a column packed with Sephadex (LH-2O) or.

Collecting the effluent fractions with chloroform-hexane (65:35 V/V) yielded 1α, 3β, and
24-trihydroxycholester-5,7-diene36.
6■ is obtained.

UV spectrum: λ””263,271° extreme α 282.294mμ Mass spectrum: m/e 416 (M+).

398.380.357 j) Ether of 1α, 3β, 24-hydroxycholester 5,7-diene 23.7η obtained in i) above 380
The solution in R111 was heated using a 400W high-pressure mercury lamp (Toshiba).
UV rays are irradiated for 1 minute in an argon gas atmosphere through a Vycor filter.

Then, the solvent was distilled off at room temperature under reduced pressure, and the residue was subjected to column chromatography packed with Sephadex (LH-20) 10F.

The oily 1α,24-dihydroxyprevitamin D3 is obtained from the first running fraction with chloroform-hexane (65:35 V/V).

This material exhibits maximum ultraviolet absorption at 260 mμ in ether solution.

k) 1α,24-dihydroxy previtamin D3 at 10
Dissolve in 0 ml of ether and leave for 2 weeks in the dark at room temperature under an argon gas atmosphere.

During this period, the maximum absorption position of ultraviolet rays is from 260 mμ to 264 m
The absorption intensity shifts to μ and increases by about 1.6 times.

The solvent was distilled off at room temperature under reduced pressure, and the residue was subjected to chromatography using a column packed with Sephadex (LH-2o) toy.

Oily 1α,24-dihydroxycholecalciferol 5.9′11Ni is obtained from the fraction runoff with chloroform-hexane (65:35 V/V).

UV spectrum: λEtoH264mμ Mass spectrum: m/e 416 (ψ-).

398.380,269,251,152°134.1
05°

Claims (1)

[Claims] 3,24-dihydroxy-cholescun represented by the formula 1 is treated with 2,3-dichloro-5,6-diciatzbenzoquinone in a solvent to obtain 24-hydroxy-cholesta-1, represented by the formula 4-dien-3-one and then 2,3-dichloro-5,6-
24-Hydroxy-cholest-1
,5,7-drien-3-one, which was reduced with metal hydride to give 3β,24-dihydroxy-cholester-1,5
, 7-triene 1 represented by the general formula (in the formula, R means an organic residue). A 1,4-cycloadduct represented by the general formula OH (wherein R means the same as above) is obtained by reacting with a 2,4-triazo so-3,5-dione derivative, which is then treated with peroxide. 1α represented by the general formula (wherein R means the same as above)
, 2α-epoxide compound, and then reduced with metal hydride to produce 1α and 3β. 1α,24-dihydroxyprevitamin D was obtained by irradiating 24-trihydroxy-cholesta-5,7-diene with ultraviolet light.
1, characterized in that it is prepared with 3 and then isomerized.
Method for producing α,24-dihydroxycholecalciferol.