JPS6148501B2 - - Google Patents

Description

[Detailed Description of the Invention] The present invention provides a novel vitamin represented by the following formula.
This article relates to _D3 derivatives and their production methods.

A compound of the formula has the general formula (In the formula, R ₁ , R ₂ and R ₃ may be the same or different,
A provitamin _D3 derivative represented by a hydrogen atom or a protecting group for a hydroxyl group, respectively, is irradiated with ultraviolet light to obtain the general formula (wherein R ₁ , R ₂ and R ₃ have the above-mentioned meanings) and then thermally isomerized to form a previtamin D ₃ derivative represented by the general formula It is produced by producing a vitamin D ₃ derivative represented by the formula (wherein R ₁ , R ₂ and R ₃ have the above-mentioned meanings) and removing the protecting group of the hydroxyl group as necessary.

As the protecting groups R ₁ , R ₂ and/or R ₃ for the hydroxyl group, common protecting groups can be used as long as they do not interfere with the reaction caused by ultraviolet irradiation and the isomerization reaction, and those that can be easily removed are preferred. . As the protecting group, an acyl group is usually used, such as a substituted or unsubstituted benzoyl group, an acetyl group, and the like.

The method of the present invention can be carried out, for example, as follows.

The reaction in the step of producing the previtamin D ₃ derivative of the formula by irradiating the provitamin D ₃ derivative of the formula with ultraviolet rays is preferably carried out in a solvent.
As the solvent, for example, saturated or unsaturated hydrocarbon solvents, ether solvents, and alcohols are used, and low boiling point solvents are preferred. Examples of the hydrocarbon solvent include hexane and octane; examples of the ether solvent include diethyl ether and tetrahydrofuran; examples of the alcohol include lower alcohols, preferably methanol, and mixtures of these solvents. The reaction is preferably carried out at a temperature around room temperature,
Moreover, it is preferable to carry out in an inert gas atmosphere such as argon. As the light source for ultraviolet irradiation, a normal one such as a low-pressure, medium-pressure, or high-pressure mercury lamp can be used. 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.

The obtained previtamin D ₃ derivative of the formula can be isolated by a conventional method, for example, by concentrating the reaction mixture and then chromatography, but it can also be used as it is in the next step without isolation.

The previtamin D ₃ derivative of the formula and the vitamin D ₃ derivative of the formula are in thermal equilibrium with each other, and the compound of the formula can be obtained by thermally isomerizing the compound of the formula. This isomerization reaction is carried out by a conventional method, for example, by leaving the compound of the formula in a solvent or heating it. As the solvent in this case, a usual solvent inert to the reaction, such as a hydrocarbon solvent, an ether solvent, an alcohol, a benzene solvent, etc., is used. Among them, those having a low boiling point and excellent solubility of the compound of the formula are preferable, and examples of examples include hexane, isooctane, benzene, toluene, diethyl ether, tetrahydrofuran, and mixtures thereof. The reaction time is appropriately selected from several minutes to several weeks depending on the reaction temperature. The isomerization reaction is preferably carried out in an inert gas atmosphere such as argon.

Isolation of the vitamin D ₃ derivative of formula from the reaction mixture can be achieved by commonly used purification means, e.g. extraction,
It is performed by recrystallization, column chromatography, preparative high performance liquid chromatography, preparative TLC, etc.

In the vitamin D ₃ derivative of the obtained formula,
When R ₁ , R ₂ and R ₃ are each a hydrogen atom, it is the same as the compound () of the present invention. When at least one of R ₁ , R ₂ and R ₃ is a hydroxyl protecting group, the compound () of the present invention is produced by further removing this group.

The removal of the protecting group for the hydroxyl group differs depending on the type of protecting group, but for example, when the protecting group is an acyl group,
Deacylation is carried out in a conventional manner, for example by treatment with a basic substance or metal hydride. In this case, the basic substance used is, for example, an alkali metal hydroxide, preferably potassium hydroxide, sodium hydroxide, etc., and the hydride metal is, for example, lithium aluminum hydride, lithium boron hydride, boron hydride, etc. Sodium etc. are commonly used. The deacylation reaction is preferably carried out in a solvent. The solvent used is appropriately selected depending on the type of deacylating agent. For example, when an alkali metal hydroxide is used, water, alcohol, or a mixture of these and another solvent such as ether is generally used. In the case of reductive deacylation using hydrogenated metals, ether solvents are suitable, such as diethyl ether, tetrahydrofuran, 1,2-dimethoxyethane, diglyme, etc., or mixtures thereof. When sodium borohydride is used, alcohols such as methanol, ethanol, etc. can also be used. The reaction temperature is appropriately selected between room temperature and the boiling temperature of the mixture depending on the type of deacylating agent, but it is 100℃ when using an alkali metal hydroxide.
The following are preferred.

Isolation and purification of the compound of the formula can be carried out by conventional methods, for example, by an appropriate combination of extraction, recrystallization, chromatography, etc.

Compounds of the formula of the present invention (24,24-difluoro-
1α·25-dihydroxy-vitamin D ₃ ) exhibits strong vitamin D-like biological activity and is useful as a medicine.

Formula provitamin used as starting material
D ₃ derivatives are new compounds, for example with the general formula It can be produced starting from a compound represented by the formula (wherein X ₁ and X ₂ represent a hydrogen atom or a carbon-carbon single bond).

As the compound of the formula, for example, 24,24-difluoro-5β-cholestane-3α,25-diol (a), 24,24-difluoro-25-hydroxycholesterol (b), etc. are used, and these can be prepared by known methods. For example, 24,24-difluoro-25
-Hydroxy-1,4-cholestadien-3-one () or 24,24-difluoro-25-hydroxy-1,4,6-cholestadien-3-one (). Note that the synthetic route for the starting material () of the present invention is a 1,4-dienone compound () or a 1,4,6-trienone compound ().
The difference is slightly different depending on the case where , and will be explained below.

(A) When using the 1,4-dienone compound (), the starting material () is produced by the following route. In the formula, Ac means an acyl group.

In this method A, the isomerization of compound () is carried out using an organic base such as potassium-tert-butoxide, etc. in a solvent such as ether, isopropyl ether, tetrahydrofuran, dimethyl sulfoxide, tertiary butyl alcohol, etc. or a mixture thereof. It is carried out by the action of a metal alkoxide. The obtained compound () is then reduced in an inert organic solvent using a metal hydride such as lithium aluminum hydride, lithium borohydride, sodium borohydride, etc. or using isopropanol-aluminum isopropoxide. The compound () is produced by subjecting it to a Mehrwein-Pondorf reduction system. Hydroxylation at the 1α position of the compound () can be carried out, for example, by a method using a mercuric salt of a strong organic acid (see JP-A-52-71456) or a method using a hydroboration reaction (JP-A-50-7145).
12069)). The compound () obtained in this way has its 1st and 3rd positions.
After protecting the free hydroxyl group at position with acyl group etc.,
By a series of steps of halogenation and dehydrohalogenation of the carbon at position 7, which is the usual means of producing a diene compound), a starting formula in which R ₁ and R ₂ are hydroxyl protecting groups and R ₃ is a hydrogen atom is obtained. Induced by substance (a).

When the obtained compound (a) is deacylated by a conventional method, a compound (b) of the formula in which R ₁ , R ₂ and R ₃ are hydrogen atoms is obtained. (In the formula, R means an organic residue) by acting with a 1,2,4-triazoline-3,5-dione derivative, A 1,4-cycloadduct represented by the formula (wherein R has the above-mentioned meaning) is produced, and then this is reductively deacylated using a hydrogenation metal such as lithium aluminum hydride. It can be manufactured even if it is twisted. The latter method has the advantage that compound (a) obtained from compound () by the above operation can be used as it is without isolating it from the reaction mixture, and it can efficiently produce compound (b) with good purity. Excellent.

(B) When using the 1,4,6-trienone compound (), the starting material () is produced by the following route. In the formula, R has the above meaning.

This method B is disclosed in Japanese Patent Application Laid-open No. 50-84560 and Japanese Patent Application Laid-open No.
It can be carried out by the method described in No. 50-84555. Compound () is obtained by acylating the compound () obtained by the above method (A).
After forming 24,24-difluoro-25-hydroxycholester-1,5-diene-acylate,
It can also be produced by the method described in JP-A-53-71055.

Compounds of the formula include, for example, lithocholic acid methyl ester, 3β-hydroxy-5-cholenic acid methyl ester, 3β-hydroxy-5-homocholenic acid lower alkyl ester, etc., as shown below.
Produced by methods (C), (D), and (E).

(C) Production of compound () from lithocholic acid methyl ester: (D) Production of compound () from 3β-hydroxy-5-cholenic acid methyl ester: (E) Production of compound () from 3β-hydroxy-5-homocholenic acid methyl ester: In each of the above methods (C), (D) and (E), 3α-acetoxy-24-oxo-5β-homocholanic acid ethyl ester, 3β-acetoxy-24-oxo-5β
- The reaction of difluorinating the 24-position of a ketocarboxylic acid such as homocholenic acid methyl ester and 3β-acetoxy-24-oxo-5β-homocholenic acid ethyl ester to produce the corresponding 24,24-difluoro compound is carried out using an inert organic solvent such as benzene. , diglyme, dichloromethane, etc. by treatment with diethylaminosulfate trifluoride. Although there is no particular restriction on the reaction temperature, preferable results can be obtained if the reaction is carried out at a low temperature. The reaction time is appropriately selected from several tens of minutes to several hours. 24.24- from the corresponding 24.24-difluoro compound obtained
The reaction for producing 25-hydroxy forms such as difluoro-5β-cholestane-3α・25-diol and 24・24-difluoro-25-hydroxycholesterol involves the reaction of methyl ester in an inert organic solvent such as tetrahydrofuran, ether, diglyme, or benzene. This is done by treatment with methyl metal such as lithium or methylmagnesium bromide.

Details of each reaction in Methods (A) to (E) are described in Reference Examples. Isolation of the target compound from the reaction mixture is
Each is carried out using conventional methods.

Reference example 1 24,24-difluoro-5β-cholestane-3α,25-diol (a) and 24,24-difluoro-25- from lithocholic acid methyl ester
Production of hydroxycholesterol (b): (a) Dissolve 970 mg of methyl lithocholic acid and 474μ of dihydropyran in 1.5 ml of dichloromethane, and dissolve p-toluenesulfonic acid pyridinium salt.
Add 62.3 mg and leave for 1 hour. Add 25 ml of dichloromethane to the reaction mixture, wash with saturated brine, dry the organic layer over anhydrous potassium carbonate, and then evaporate the solvent. The resulting colorless oily substance was subjected to column chromatography (15 g of silica gel,
When purified by hexane/ethyl acetate = 10:1), 3-O-(2-tetrahydropyranyl)-
1165 mg of lithocholic acid methyl ester () are obtained as a colorless oil.

IR spectrum ν _nax (cap): 1730 cm ^-1 NMR spectrum δ (CDCl ₃ ): 0.63 (3H,
s), 0.90 (3H, s), 3.2~4.2 (3H, m),
3.33 (3H, s), 4.72 (1H, m, W/2 = 6
Hz) (b) 32 mg of lithium aluminum hydroxide is suspended in 1.5 ml of tetrahydrofuran, and 3-O-(2
A solution of 389 mg of methyl ester (-tetrahydropyranyl)-lithocholic acid is added dropwise at 0 DEG C. while stirring under a stream of argon. After stirring for 10 minutes at 0°C and then for 30 minutes at room temperature, the excess reagent is decomposed and removed by adding dropwise 0.15 ml of water with stirring at 0°C. The liquid was concentrated under reduced pressure, and the resulting colorless oil was purified by column chromatography (6 g of silica gel, hexane/ethyl acetate = 10:3) to give 3α-(2-tetrahydropyranyloxy)-24- 358 mg of hydroxy-5β-cholane are obtained as a white powder.

IR spectrum ν _nax (CHCl ₃ ): 3480 cm ^-1 NMR spectrum δ (CDCl ₃ ): 0.65 (3H,
s), 0.90 (3H, s), 3.2~4.0 (5H, m),
4.75 (1H, m, w/2 = 6Hz) (c) 221 mg of pyridium chlorochromate (C ₅ H ₅ N ⁺ HCrO ₃ Cl ^- ) and 8.3 mg of sodium acetate were suspended in 1 ml of dichloromethane, and to this suspension, 3α dissolved in 4 ml of dichloromethane
A solution of 221 mg of -(2-tetrahydropyranyloxy)-24-hydroxy-5β-cholane is added in one portion under stirring under a stream of argon and stirred for 3 hours. The supernatant of the reaction mixture is decanted and the precipitate is washed three times with dichloromethane. The supernatant and washing liquid were combined, passed through a silica gel (4 g) shoto column, washed with dichloromethane, and the passed liquid was concentrated under reduced pressure to yield 3α-(2-tetrahydropyranyloxy)-24-oxo-5β-cholane 177
mg as a colorless oil.

IR spectrum ν _nax (CHCl ₃ ): 1715 cm ^-1 NMR spectrum δ (CDCl ₃ ): 0.65 (3H,
s), 0.97 (3H, s), 3.3-4.2 (3H, m),
4.75 (1H, m, w/2=6Hz), 9.82 (1H,
t, J=2Hz) (d) 3α-(2-tetrahydropyranyloxy)-
19.3 g of 24-oxo-5β-cholane and 1,3-
8.92ml of propanediol to 100ml of dichloromethane
ml and stirred at −78 °C under a stream of argon.
Then add 560μ of boron trifluoride/diethyl ether and stir for 15 minutes. After further stirring at room temperature for 3 hours, the reaction mixture was diluted with 10% potassium hydroxide solution,
The organic layer is washed successively with water and saturated brine, dried over anhydrous magnesium sulfate, and the solvent is distilled off. Dissolve the residue in 15 ml of ethyl acetate and add hexane.
Add 30ml, leave it at -10℃ for 4 hours, and remove the precipitated crystals.3α-hydroxy-24・24
9.6 g of -propylene dithio-5β-cholan are obtained as colorless needles.

Melting point: 145-146℃ (hexane/ethyl acetate =
(Recrystallized from 10:3) IR spectrum ν _nax (KBr): 3300 cm ^-1 NMR spectrum δ (CDCl ₃ ): 0.63 (3H,
s), 0.92 (3H, s), 2.5-3.1 (4H, m),
3.60 (1H, m, w/2 = 15Hz), 3.98 (1H,
t, J = 6 Hz) (e) 4.8 g of 3α-hydroxy-24,24-propylene dithio-5β-chorane and dihydropyran
Dissolve 1.53 ml in 15 ml of dichloromethane, add 266 mg of p-toluenesulfonic acid pyridinium salt, and leave for 2 hours. 50 ml of dichloromethane is added to the reaction mixture, and the organic layer is washed with saturated brine and dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent was purified by column chromatography (120 g of silica gel, hexane/ethyl acetate = 10:1) to obtain 3α-(2
3.5 g of -tetrahydropyranyloxy)-24.24-propylene dithio-5β-choran are obtained as a white powder.

(f) 3α-(2-tetrahydropyranyloxy)-
24・24-propylene dithio-5β-cholan 12g
was dissolved in 40 ml of tetrahydrofuran, and 16.4 ml of a hexane solution containing 15% n-butyllithium was added dropwise at -20° C. under stirring in an argon stream.
After stirring for 2 hours, the reaction mixture was poured into 43 ml of ethyl chlorocarbonate under stirring at -78°C under an argon atmosphere.
Add dropwise over 10 minutes and continue stirring at room temperature for 2 hours. The reaction mixture was filtered, the liquid was concentrated under reduced pressure,
The obtained oily residue was subjected to column chromatography (1200 g of silica gel, hexane/ethyl acetate =
10:1), 1.8 g of raw material and 3α-(2-tetrahydropyranyloxy)-
8.85 g of 24.24-propylene dithio-5β-homocholanic acid ethyl ester are obtained as a white powder.

IR spectrum ν _nax (CHCl ₃ ): 1720 cm ^-1 NMR spectrum δ (CDCl ₃ ): 0.63 (3H,
s), 0.91 (3H, s) 1.34 (3H, t, J = 6.5
Hz), 4.27 (2H, q, J = 6.5Hz), 4.74
(1H, m, w/2 = 6Hz) (g) 3α-(2-tetrahydropyranyloxy)-
Dissolve 493 mg of 24,24-propylene dithio-5β-homocholanic acid ethyl ester in 10 ml of ethanol, and add p-toluenesulfonic acid pyridium salt 21
mg and stirred at 55°C for 3 hours. The reaction mixture was concentrated under reduced pressure, and the residue was subjected to column chromatography (10 g of silica gel, hexane/ethyl acetate =
5:2), 428 mg of 3α-hydroxy-24·24-propylene dithio-5β-homocholanic acid ethyl ester is obtained as colorless crystals.

IR spectrum ν _nax (CHCl ₃ ): 1715 cm ^-1 NMR spectrum δ (CDCl ₃ ): 0.64 (3H,
s), 0.93 (3H, s), 1.34 (3H, t, J=
7 Hz), 4.28 (2H, q, J = 7 Hz) (h) 3α-hydroxy-24,24-propylene dithio-5β-homocholanic acid ethyl ester 428 mg
was dissolved in 5 ml of pyridine, 2 ml of acetic anhydride was added dropwise with stirring at 0°C, and the mixture was left at room temperature for 14 hours. The reaction mixture was poured into 30 ml of 2N hydrochloric acid and extracted with ethyl acetate. The organic layer is washed successively with 2N hydrochloric acid, water, saturated aqueous sodium bicarbonate solution, and saturated brine, and dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent was purified by column chromatography (120 g of silica gel, hexane/ethyl acetate = 5:1) to obtain 3α-acetoxy-24·24-propylene dithio-5β-
417 mg of homocholanic acid ethyl ester are obtained as colorless crystals.

IR spectrum ν _nax (CHCl ₃ ): 1715, 1720
(sh) cm ^-1 NMR spectrum δ (CDCl ₃ ): 0.64 (3H,
s), 0.93 (3H, s), 1.34 (3H, t, J=
7Hz), 2.04 (3H, s), 2.3~3.2 (4H,
m), 4.30 (2H, q, J=7Hz), 4.73
(1H, m, w/2 = 20Hz) (i) Dissolve 847 mg of N-bromosuccinimide in 20 ml of 95% acetone water, and add 3α-acetoxy-24.24 dissolved in 30 ml of 95% acetone water to this solution.
A solution of 336 mg of -propylene dithio-5β-homocholanic acid ethyl ester is added under stirring at 0° C. and stirred for 15 minutes. The reaction mixture was poured into 50 ml of saturated acidic sodium sulfite aqueous solution and diluted with hexane/
Extract with a mixture of dichloromethane (1:1),
The separated organic layer was dissolved in saturated aqueous sodium bicarbonate solution.
Wash twice with 30 ml and once with 50 ml of saturated saline. These washings are again extracted twice with 20 ml of a mixture of hexane/dichloromethane (1:1) and washed with saturated brine. The separated organic layers are combined, washed again with saturated brine, and dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent was purified by column chromatography (hexane/ethyl acetate = 6:1) to obtain 3α-acetoxy-24-oxo-5β.
-254 mg of homocholanic acid ethyl ester are obtained as colorless crystals.

IR spectrum ν _nax (CHCl ₃ ): 1720 cm ^-1 NMR spectrum δ (CDCl ₃ ): 0.64 (3H,
s), 0.94 (3H, s), 1.37, (3H, t, J=
7Hz), 2.04 (3H, s), 1.78 (2H, bs,
w/2=15Hz), 4.34 (2H, q, J=7
Hz), 4.78 (1H, m, w/2 = 20Hz) (j) Diethylaminosulfate trifluoride produced from N.N-diethylaminotrimethylsilane and sulfur tetrafluoride [(C ₂ H ₅ ) ₂ NSF ₃ ] Dissolve 3.3 ml in 20 ml of dichloromethane, and add to this solution 3α-acetoxy-24-oxo-5β- dissolved in 60 ml of dichloromethane.
A solution of 5.5 g of homocholanic acid ethyl ester,
The mixture was added at −78° C. under stirring in an argon atmosphere and left at room temperature for 3 hours. The reaction mixture is poured into a mixture of 200 ml of saturated aqueous sodium bicarbonate solution and 150 ml of hexane and stirred vigorously, and the separated organic layer is washed successively with saturated aqueous sodium bicarbonate solution and saturated brine, and dried over anhydrous magnesium sulfate. . The residue obtained by distilling off the solvent was purified by column chromatography (100 g of silica gel, hexane/ethyl acetate = 10:1), and 700 mg of raw material was recovered, and 3α-acetoxy-24,24-difluoro-5β- 3.7 g of homocholanic acid ethyl ester are obtained as a colorless oil.

IR spectrum ν _nax (CHCl ₃ ): 1755, 1720 cm
^-1 NMR spectrum δ (CDCl ₃ ): 0.65 (3H,
s), 0.93 (3H, s), 1.36 (3H, t, J=
7Hz), 2.03 (3H, s), 4.35 (2H, q, J
= 7 Hz), 4.73 (1H, m, w/2 = 20 Hz) (k) 3α-acetoxy-24,24-difluoro-5β- dissolved in 1 ml of tetrahydrofuran in 970μ of a solution of methylmagnesium bromide in tetrahydrofuran (1 mol/) A solution of 60 mg of homocholanic acid ethyl ester is added dropwise at 0° C. under stirring under a stream of argon and stirred for 30 minutes at room temperature. Add 10% of 2N hydrochloric acid to the reaction mixture under stirring at 0°C.
ml was added dropwise, stirred for 5 minutes, and then extracted with ethyl acetate. The separated organic layer was treated with 2N hydrochloric acid, water,
Wash sequentially with saturated aqueous sodium bicarbonate solution and saturated brine, and dry over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent was subjected to column chromatography (10 g of silica gel, hexane/
When purified with ethyl acetate = 1:1), 24.
24-difluoro-5β-cholestane-3α・25
-39 mg of diol (a) are obtained as colorless crystals.

Melting point: 142-142.5℃ (hexane/ethanol =
(recrystallized from 50:1) IR spectrum ν _nax (CHCl ₃ ): 3620, 3450 cm
^-1 NMR spectrum δ (CDCl ₃ ): 0.66 (3H,
s), 0.92 (3H, s), 1.31 (6H, s), 3.65
(1H, m, w/2 = 18Hz) Mass spectrum m/e: 440 (M ⁺ ), 423,
422 (base peak) (l) 24・24-difluoro-5β-cholestane-3
Dissolve 800mg of α-25-diol (a) in 32ml of tertiary butyl alcohol, 15ml of water, 200μ of 48% hydrobromic acid and 800mg of N-bromoacetamide.
and stirred at room temperature for 3 days. Ethyl acetate is added to the reaction mixture, washed with 0.5% aqueous sodium thiosulfate solution, then saturated brine, and dried over anhydrous magnesium sulfate. The residue obtained by evaporation of the solvent was purified by passing it through a Schiot column containing 5 g of silica gel to obtain 950 mg of a crude product, which was recrystallized from ether-hexane to give 4.
β-bromo-24·24-difluoro-25-hydroxy-5β-cholestan-3-one is obtained.

Melting point: 152-153℃ NMR spectrum δ ( _CDCl3 ): 0.70 (3H,
s), 1.10 (3H, s), 1.30 (6H, s), 5.00
(1H, d, J = 13Hz) Mass spectrum m/e: 519, 517 (M ⁺ ), 438 (m) 4β-bromo-24・24-difluoro-25-
Hydroxy-5β-cholestan-3-one 950
mg is dissolved in 15 ml of dimethylformamide, 500 mg of lithium carbonate is added, and the mixture is heated under reflux for 2 hours while stirring. After cooling, about 100 ml of water was added to the reaction mixture, and the precipitated crystals were filtered, washed with water, and dried to obtain 730 mg of the crude enone, which was subjected to column chromatography (silica gel 40 g, hexane/ethyl acetate = 7 :3), 24,24-difluoro-25-hydroxy-4
-498 mg of cholesten-3-one are obtained as crystals.

Melting point: 177-178℃ (recrystallized from ether-hexane) NMR spectrum δ (CDCl ₃ ): 0.70 (3H,
s), 1.16 (3H, s), 1.32 (6H, s), 5.77
(1H, s) Mass spectrum m/e: 436 (M ⁺ ), 394 (n) 24・24-difluoro-25-hydroxy-4
-457 mg of cholesten-3-one in 46 ml of ether
and 650 ml of dimethyl sulfoxide, and further stirred at 10°C for 15 minutes. The reaction mixture was poured into ice water saturated with carbon dioxide gas, stirred vigorously, and then extracted with ethyl acetate. The separated organic layer is washed with water, dried over anhydrous magnesium sulfate, and the solvent is distilled off to obtain 24.24-difluoro-25-hydroxy-5-cholesten-3-one as crude crystals.

NMR spectrum δ( _CDCl3 ): 0.70(3H,
s), 1.20 (3H, s), 1.30 (6H, s), 5.35
(1H, m) Without purifying this product, add 13 ml of ether.
and 25 ml of methanol, and add 10 ml of an aqueous solution of 200 mg of sodium borohydride at 0°C.
Stir at 0°C for 1 hour. The reaction mixture is extracted with ethyl acetate, and the separated organic layer is washed with 1% hydrochloric acid and saturated brine, and dried over anhydrous magnesium sulfate. The residue obtained by distillation with a solvent was purified by column chromatography (15 g of silica gel, hexane/ethyl acetate = 2:1) to obtain 24,24-difluoro-25-hydroxycholesterol (
b) 320 mg are obtained as crystals.

Melting point: 168-170℃ (recrystallized from ether) NMR spectrum δ (CDCl ₃ ): 0.70 (3H,
s), 0.95 (3H, d, J=6Hz), 1.02 (3H,
s), 1.32 (6H, s), 5.40 (1H, d, J=4
Hz) Mass spectrum m/e: 438 (M ⁺ ), 420 Reference example 2 3β-acetoxy-24-oxo-5- from 3β-hydroxy-5-cholenic acid methyl ester
Production of homocholenic acid ethyl ester: When 3β-hydroxy-5-cholenic acid methyl ester is treated in the same manner as described in Reference Example 1(a) to (i), 3β-acetoxy-24-oxo-5
- Homocholenic acid ethyl ester is obtained.

Reference example 3 3β-acetoxy-24-oxo- from 3β-hydroxy-5-homocholenic acid methyl ester
Production of 5-homocholenic acid methyl ester: Dissolve 4 mmol of diisopropylamine in 5 ml of tetrahydrofuran, add 1.92 ml of a hexane solution containing 4 mmol of n-butyllithium at 0°C with stirring in an argon stream, and stir at 0°C for 10 minutes. Stir. 3β-
(2-tetrahydropyranyloxy)-5-homocholenic acid methyl ester (3β-hydroxy-5-
2.5 ml of a 4 mmol tetrahydrofuran solution (obtained from homocholenic acid methyl ester in the same manner as in Reference Example 1(a)) was added dropwise over 15 minutes. After stirring the reaction mixture at -78°C for 30 minutes, 4 mmol of phenyl disulfide was added to 2 ml of tetrahydrofuran.
A mixture of 4 mmol of hexamethylphosphoramide and hexamethylphosphoramide was added dropwise over 20 minutes, and the mixture was further stirred at -78°C for 30 minutes. Gradually warm to room temperature and add 40 ml of water to the reaction mixture.
ml and extract with ether. 10% organic layer
Washed sequentially with an aqueous sodium hydroxide solution, water, 10% hydrochloric acid and water, dried over anhydrous magnesium sulfate,
When the solvent was distilled off, 3β-(2-tetrahydropyranyloxy)-24,24-diphenylthio-5-
Homocholenic acid methyl ester is obtained. Subsequently, this 24,24-diphenylthio compound was added to Reference Example 1 (g) and
When treated in the same manner as (h), 3β-acetoxy-24.
24-diphenylthio-5-homocholenic acid methyl ester is obtained. The thus obtained 3β-acetoxy-24·24-diphenylthio-5-homocholenic acid methyl ester was heated to reflux in a methanolic iodine solution and then treated with an aqueous trifluoroacetic acid solution, resulting in 3β-acetoxy-24-oxo-5-
Homocholenic acid methyl ester is obtained.

Reference Example 4 3β-acetoxy-24-oxo-5-homocholenic acid ethyl ester obtained in Reference Example 2 or 3β-acetoxy-24-oxo- obtained in Reference Example 3
When 5-homocholenic acid methyl ester was treated in the same manner as in Reference Example 1(j) and (k), 24,24-difluoro-
Crystals of 25-hydroxycholesterol (b) are obtained.

This product did not show any melting point depression in the mixing test with the compound obtained in Reference Example 1(n), and
The NMR spectrum and mass spectrum also matched.

Reference example 5 24・24-difluoro-5β-cholestane-3
24・24-difluoro from α・25-diol
Production of 5,7-cholestadiene-1α,3β,25-triol: (a) 24,24-difluoro-5β-cholestane-3
100 mg of α·25-diol was dissolved in 2 ml of dioxane, 207 mg of 2,3-dichloro-5,6-dicyanobenzoquinone was added, and the mixture was heated under reflux for 4 hours.
After cooling, filter the reaction solution and add 4g of alumina to the solution.
Pass through a Schiot column and elute with ethyl acetate/ethanol (2:1), and concentrate the eluate under reduced pressure. The resulting residue was purified by preparative thin layer chromatography using 20 g of silica gel (hexane/ethyl acetate = 5:2, developed three times) to yield 24,24-difluoro-25-hydroxy-1,4-cholestadiene. -3-one ()
45 mg are obtained as needles.

Melting point: 176-177℃ (hexane/ethyl acetate =
Recrystallized from 1:1) IR spectrum ν _nax (KBr): 3470, 1660,
1615cm ^-1 UV spectrum λ _nax (C ₂ H ₅ OH): 247,
296nm NMR spectrum δ (CDCl ₃ ): 0.75 (3H,
s), 1.30 (6H, s), 1.57 (3H, s), 6.08
(1H, s), 6.25 (1H, dd, J=10Hz), 2
Hz), 7.03 (1H, d, J = 10Hz) (b) 24・24-difluoro-25-hydroxy-1・
4-cholestadien-3-one () 513mg
was dissolved in a mixture of 15 ml of dimethyl sulfoxide and 5 ml of ether, and 662 mg of potassium tert-butoxide was added at 15°C under stirring in an argon stream.
Add and continue stirring for 30 minutes. Water the reaction mixture
Pour into a mixture of 90 ml and 5 g of dry ice, stir, and extract with ether. The separated organic layer is washed with saturated brine and then concentrated under reduced pressure. The resulting residue was mixed with 5 ml of ether and 25 ml of methanol.
ml of the mixture, add 5 ml of an aqueous solution of 25 mg of sodium borohydride under stirring at 0°C, and stir for 15 minutes. The reaction mixture was poured into 60 ml of 2N HCl and extracted with ethyl acetate. The separated organic layer is washed with saturated brine and then dried over anhydrous potassium carbonate. The residue (474 mg) obtained by distilling off the solvent was purified by column chromatography (20 g of silica gel, hexane/ethyl acetate = 2:1) to obtain 24,24-difluoro-1,5.
-cholestadiene-3β・25-diol ()
252 mg are obtained as white powder crystals.

Melting point: 170-171℃ (hexane/ethyl acetate =
Recrystallized from 4:1) IR spectrum ν _nax (KBr): 3400, 3550cm
^-1 NMR spectrum δ (CDCl ₃ ): 0.74 (3H,
s), 1.12 (3H, s), 1.34 (6H, s), 4.22
(1H, m, W/2 = 18Hz), 5.4~5.5 (1H,
m), 5.59 (1H, d, J = 10Hz), 5.83
(1H, dd, J = 10Hz, 2Hz) Mass spectrum m/e: 436 (M ⁺ , base peak), 421, 418, 407, 403 (c) Dissolve 445 mg of mercuric acetate in 1.4 ml of water and store at room temperature. Add 1.4 ml of tetrahydrofuran and 108μ of trifluoroacetic acid while stirring, and after 1 minute add tetrahydrofuran/dimethylformamide (1:1).
A solution of 122 mg of 24,24-difluoro-1,5-cholestadiene-3β,25-diol () dissolved in 1.4 ml of the mixed solution was added, and the mixture was stirred for 2.5 hours. Add 2 ml of 2N aqueous sodium hydroxide solution to the reaction mixture, and add sodium borohydride dissolved in 0.7 ml of 2N aqueous sodium hydroxide solution.
Add 16 mg of solution and stir for 1 hour. Add 2.5 g of potassium carbonate to the reaction mixture, separate the organic layer, extract the aqueous layer five times with 15 ml of tetrahydrofuran, and combine the organic layer and the extract. The residue obtained by concentration under reduced pressure is dissolved in 30 ml of chloroform, washed with saturated brine, and dried over anhydrous magnesium sulfate. 15 ml of benzene is added to the residue obtained by distilling off the solvent, and water is removed azeotropically. This operation is repeated five times and the residue is dried in a desiccator overnight to obtain 125 mg of a colorless solid. This solid was dissolved in 1 ml of pyridine, 1 ml of acetic anhydride was added with stirring at 0°C, and the mixture was left overnight.
The reaction mixture was poured into 30 ml of 2N hydrochloric acid, extracted with ethyl acetate, and the separated organic layer was poured into 2N hydrochloric acid,
Wash successively with water, saturated aqueous sodium bicarbonate and saturated brine, and dry over anhydrous sodium sulfate. The colorless oil (130 mg) obtained by distilling off the solvent was purified by column chromatography (20 g of silica gel, hexane/ethyl acetate = 5:2), and 24 mg of the raw material 3-acetate was recovered, and 1α/3β- Diacetoxy-24・24
-difluoro-5-cholesten-25-ol 40
mg is obtained as an oil.

NMR spectrum δ( _CDCl3 ): 0.67(3H,
s), 0.91 (3H, d, J=5Hz), 1.30
(6H, s), 2.03 (3H, s), 2.08 (3H,
s), 4.90 (1H, m), 5.08 (1H, t, J=
2Hz), 5.22 (1H, d, J = 4Hz) Mass spectrum m/e: 538 (M ⁺ ), 436,
418 (base peak) (d) Dissolve 33 mg of 1α・3β-diacetoxy-24・24-difluoro-5-cholesten-25-ol in 1 ml of hexane/benzene (1:1), and dissolve N-
Add 13.1 mg of bromosuccinimide and heat under reflux for 30 minutes in an argon atmosphere. After cooling, the reaction mixture was filtered, the liquid was concentrated under reduced pressure, the resulting residue was dissolved in 1 ml of xylene, and 85μ of s-collidine was added.
and heat to reflux for 30 minutes in an argon atmosphere. After cooling, the reaction mixture was poured into 20 ml of 2N hydrochloric acid and extracted with ethyl acetate. The separated organic layer
Wash sequentially with 2N hydrochloric acid, water, saturated aqueous sodium bicarbonate and saturated brine, and dry over anhydrous magnesium sulfate. The pale yellow oil obtained by distilling off the solvent (32 mg) was dissolved in 1 ml of ether, 7.5 mg of 4-phenyl-1,2,4-triazoline-3,5-dione (XI) was added, and the mixture was heated under an argon stream. Leave it overnight at room temperature. The reaction mixture was subjected to preparative thin layer chromatography using 10 g of silica gel (hexane/ethyl acetate =
1:1), 1α·3β-diacetoxy-24·24-difluoro-5·7-cholestadien-25-ol and 4-phenyl-1·
12 mg of 1,4-cycloadduct (XII) with 2,4-triazoline-3,5-dione are obtained as crystals.

IR spectrum ν _nax (CHCl ₃ ): 1730, 1690 cm
^-1 NMR spectrum δ (CDCl ₃ ): 0.80 (3H,
s), 1.27 (6H, s), 1.30 (3H, s), 1.99
(3H, s), 210 (3H, s), 6.30 (1H, d,
J = 9 Hz), 6.45 (1H, d, J = 9 Hz) (e) Suspend 6.4 mg of lithium aluminum hydride in 0.5 ml of tetrahydrofuran, and add to this suspension,
Reference example 5(d) dissolved in 1 ml of tetrahydrofuran
A solution of 12 mg of the 1.4-cycloadduct (XII) obtained in step 1 is added dropwise while heating under reflux in an argon stream. After leaving for 2 hours, add water at 0°C with stirring.
Add 0.5 ml dropwise and pour into 2N hydrochloric acid. Extract with ethyl acetate, and add the separated organic layer to 2N-hydrochloric acid,
Wash successively with water, saturated aqueous sodium bicarbonate and saturated brine, and dry over anhydrous sodium sulfate. The residue (12 mg) obtained by distilling off the solvent was purified by preparative thin layer chromatography (hexane/ethyl acetate = 1:2).
3 mg of 24,24-difluoro-5,7-cholestadiene-1α,3β,25-triol (b) are obtained as colorless crystals.

IR spectrum ν _nax (CHCl ₃ ): 3600, 3400cm
^-1 UV spectrum λ _nax (EtOH): 262, 272,
282.5, 294nm NMR spectrum δ (CDCl ₃ ): 0.63 (3H,
s), 0.97 (3H, d, J=5Hz), 0.95
(3H, s), 1.31 (6H, s), 3.76 (1H, m,
W/2=7Hz), 4.06 (1H, m, W/2=30
Hz), 5.40 (1H, dd, J=5, 3Hz), 5.74
(1H, d, J = 5Hz) Mass spectrum m/e: 452 (M ⁺ , base peak) (f) Obtained 24,24-difluoro-5,7-cholestadiene-1α,3β,25-triol (
b) Acetylation of 5 mg with pyridine and acetic anhydride in a conventional manner gives 1α·3β-diacetoxy-24·24-difluoro-5·7-cholestadien-25-ol (a).

UV spectrum λ _nax (EtOH): 262, 272,
282, 294 nm Example 1 3 mg of 24,24-difluoro-5,7-cholestadiene-1α,3β,25-triol (b) was dissolved in 300 ml of ether and passed through a Vycor filter at 0°C with stirring in an argon stream at 200W. Irradiate with light using a high-pressure mercury lamp. The reaction mixture was concentrated under reduced pressure at 20°C, and the resulting residue was purified by preparative high performance liquid chromatography to produce 24,24-difluoro-1α,25-dihydroxyprevitamin D ₃
[UV spectrum λ _nax (EtOH): 261 nm] 370
μg is obtained. Then this previtamin D ₃ 370
Dissolve μg in 2 ml of benzene and under argon stream.
Heat to reflux for 2.5 hours. The reaction mixture was concentrated under reduced pressure at 20°C, and the resulting residue was purified by preparative high-performance liquid chromatography to recover 28 μg of previtamin D ₃ as a raw material and 24,24-difluoro-
172 μg of 1α·25-dihydroxyvitamin D ₃ is obtained.

UV spectrum λ _nax (EtOH): 266 nm NMR spectrum δ (CDCl ₃ ): 0.56 (3H,
s), 0.95 (3H, d, J=5Hz), 1.31 (6H,
s), 4.20 (1H, m), 4.40 (1H, m), 5.02
(1H, bs), 5.34 (1H, bs), 6.05 (1H, d,
J=11Hz), 6.38 (1H, d, J=11Hz) Mass spectrum m/e: 452 (M ⁺ ), 134 Example 2 1α・3β-diacetoxy-24・24 obtained in Reference Example 5(f) 5 mg of -difluoro-5,7-cholestadien-25-ol (a) was subjected to light irradiation and isomerization reaction in the same manner as in Example 1, and the obtained 1
Dissolve α-acetoxy-24,24-difluoro-25-hydroxyvitamin D ₃ -3-acetate in 5 ml of ethanol and 50μ of 25% potassium hydroxide aqueous solution.
and heat under reflux for 30 minutes to deacetylate. After cooling, the reaction mixture is poured into a mixture of chloroform and water, and the separated organic layer is washed with water and dried over anhydrous sodium sulfate. When the residue obtained by distilling off the solvent was purified by preparative high performance liquid chromatography, 24・24-difluoro-1α・
198 μg of 25-dihydroxyvitamin D ₃ is obtained. The UV and NMR spectra of this product were consistent with the compound obtained in Example 1.

Claims (1)

[Claims] Linear formula Vitamin _D3 derivative represented by. 2 General formula (In the formula, R ₁ , R ₂ and R ₃ may be the same or different,
A propitamine _D3 derivative represented by a hydrogen atom or a protecting group for a hydroxyl group, respectively, is irradiated with ultraviolet rays to obtain the general formula (wherein R ₁ , R ₂ and R ₃ have the above-mentioned meanings) and then thermally isomerized to form a previtamin D ₃ derivative represented by the general formula (wherein R ₁ , R ₂ and _{R 3} have the above-mentioned meanings) is produced, and if necessary, the protecting group of the hydroxyl group is removed.
The following formula A method for producing a vitamin _D3 derivative represented by