JPS6244558B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to novel compounds, 24,24-difluoro-25-hydroxycholestenes.

24,24-difluoro-25- provided by the present invention
Hydroxycholestenes are represented by the following formula [].

[In the formula, R is a hydrogen atom or a hydroxyl group-protecting group. Moreover, 〓 represents a single bond or a double bond. ] The compound represented by the above formula [ ] is 24,24-
It is an intermediate and a useful compound that can be derived from difluoro-25-hydroxyvitamin _D3 . That is, 24,24-difluoro-25-hydroxyvitamin D ₃ is a compound that is fully expected to exhibit the same type of pharmacological action as the vitamin D ₃ series compounds, as separately proposed by the present inventor.

Therefore, as the protecting group for the hydroxyl group in the above formula [], in the known production method of vitamin D ₃ produced from the corresponding 5-ene or 5,7-diene, the 5-ene or the 5,7- Any group that can be used as a protecting group for the hydroxyl group of a diene and can be removed by a conventional hydrolysis reaction or reductive elimination reaction may be used.

Such protecting groups are described, for example, in US Pat. No. 4,022,891.
No. stated in the specification.

Among these, particularly preferred protecting groups are acyl groups such as acetyl, benzoyl, and pivaloyl groups, and ethoxyethyl, methoxymethyl, tetrahydropyranyl, trimethylsilyl, and t-butyldimethylsilyl groups. It combines with the oxygen atom of a hydroxyl group to form an ether group.

Therefore, 24, 24- expressed by the above formula []
Difluoro-25-hydroxycholestenes are B
Depending on the number of double bonds present in the ring, the following formula []
It is divided into two groups of compounds represented by the formula -a and formula []-b.

That is, one is the following formula []-a, [In the formula, the definition of R is the same as above. ] 24,24-difluoro-25-hydroxycholest-5-ene or its hydroxy protected derivative represented by the following formula []-b, [In the formula, the definition of R is the same as above. ] 24,24-difluoro-25-hydroxycholester-5,7-diene or its hydroxyl protected derivative.

The 24,24-difluoro-25-hydroxycholestenes of the present invention can be divided into the above two groups and described as follows.

Compounds belonging to formula []-a include 24,24-difluoro-25-hydroxycholest-5-ene, 24,24-difluoro-25-hydroxycholest-5-ene-3β-acetate, 24,24 -difluoro-25-hydroxycholest-5-ene-3β-benzoate, 24,24-difluoro-25-hydroxycholest-5-ene-3β-tetrahydropyranyl ether, 24,24-difluoro-25-hydroxycholesta -5-ene-3β-trimethylsilyl ether, 24,24-difluoro-25-hydroxycholest-5-ene-3β-t-butyl dimethyl ether, and the like.

In addition, as compounds belonging to formula []-b,
24,24-difluoro-25-hydroxycholester
5,7-diene, 24,24-difluoro-25-hydroxycholester-5,7-diene-3β-acetate, 24,24-difluoro-25-hydroxycholester-5,7-diene-3β-benzoate, 24,24-difluoro-25-hydroxycholester-5,7-diene-3β-tetrahydropyranyl ether, 24,24-difluoro-25-hydroxycholester-5,7-diene-3β-t-butyldimethylsilyl ether , etc. can be given.

The above formula [] (formula []-a and formula []-
24,24-difluoro-25 represented by
-Hydroxycholestenes are produced by the following method.

That is, the following formula [], [In the formula, the definitions of R and 〓 are the same as in the case of the above formula []. ] 3β-hydroxy-24,24-difluoro-27-norcholesten-25-ones represented by methylmagnesium halide, e.g. methylmagnesium iodide, methyl React with magnesium bromide, methylmagnesium chloride, etc.

The starting material represented by the above formula [] corresponds to two compound groups, such that the above formula [] representing the target product is divided into the above formulas []-a and []-b. , that is, the following formula []-a, [In the formula, the definition of R is the same as above. ] 3β-hydroxy-24,24-difluoro-27-norcholest-5-en-25-one or its hydroxy protected derivative and the following formula []-
b, [In the formula, the definition of R is the same as above. ] 3β-hydroxy-24,24-difluoro-27-norcholester-5,7-diene-25-
ion or its hydroxyl-protected derivative.

The above formula [] (formula []-a or formula []-b)
In accordance with the definition of the group R, the compounds represented by the above formula [] are also obvious from the specific examples of the compounds represented by the above formula [].

Furthermore, the method for producing the compound of the present invention is the Grignard reaction, which is known as a reaction known per se, in an aprotic inert solvent, preferably anhydrous diethyl ether.
It is carried out under ice cooling. One of the starting materials is 3β-hydroxy-24,24- represented by the above formula []
It is generally carried out by adding an excess of methylmagnesium iodide, which is the other starting material, to difluoro-27-norcholesten-25-ones.

According to this method, as a result of the Grignard reaction, when a compound represented by the above formula [] in which R is a hydrogen atom or an acyl group is used, a target product in which R is a hydrogen atom in the formula [] can be obtained. In addition, when using a starting material represented by the above formula [] in which R is a group that combines with the oxygen atom of a hydroxyl group to form an ether group, R does not change as it is in formula []. The object expressed by the formula [] is obtained.

The thus obtained target product represented by the above formula [] is a group in which the group R forms a hydrogen atom or an ether group.

Therefore, a compound in which the group R is a hydrogen atom, that is, a compound of the above formula [] having a free hydroxyl group, is then subjected to a hydroxyl group-protecting reaction which is known per se, so that the group R is a hydroxyl-protecting group. A compound represented by the formula [] can be obtained.

Furthermore, by subjecting a compound of formula [] in which group R is a hydroxyl group-protecting group to a protective group elimination reaction known per se, a compound represented by formula [] in which group R is a hydrogen atom can be obtained. can.

The above-mentioned protection reaction of the hydroxyl group can be carried out, for example, when protecting with an acyl group, by treatment with an acid anhydride or an acyl halide in the presence of a basic compound (e.g., pyridine, etc.).
Further, when protecting with an ether group, it can be carried out by treatment with dihydropyran or methyl vinyl ether.

In addition, the elimination reaction of the protecting group of the hydroxyl group is, for example,
When the protecting group is an acyl group, this is carried out by treatment with an alkali metal hydroxide (caustic soda, caustic potash, etc.) in an alcohol solvent (ethanol, methanol, etc.), and when the protecting group is an ether group, lithium This is carried out by reduction treatment with a reducing agent such as aluminum hydride.

Note that the hydroxyl group at position 25 of the compound of the present invention can also be retained or the protective group removed by the same method as described above, if necessary.

As mentioned above, when a starting material represented by the above formula [] protected by a free hydroxyl group or a hydroxyl acyl group is used, a compound of the formula [] having a free hydroxyl group is formed by reaction with methylmagnesium halide. In this case, it can be separated from the reaction system as it is, but it can be separated from the reaction system as a compound of the above formula [] with a protected hydroxyl group by subsequently subjecting it to the above hydroxyl group protection reaction in the reaction system. It is convenient to separate them.

The target product of the present invention represented by the above formula [] can be isolated from the reaction mixture by, for example, column chromatography, thin layer chromatography, high performance liquid chromatography, recrystallization, or the like.

24,24-difluoro- provided by the present invention
25-Hydroxycholestenones are new compounds and can be easily converted into 24,24-difluoro-25-hydroxyvitamin D ₃ or its hydroxy-protected derivatives by methods known per se.

Hereinafter, the present invention will be described in detail with reference to Examples, but the 3β-hydroxy-24,24-difluoro-27-norcholesten-25-ones represented by the above formula [] or its hydroxyl-protected derivatives used in the present invention may also be used. , is a new compound as far as the inventors know,
Therefore, first, as a reference example, a method for synthesizing the compound represented by the above formula [] will be described.

In the Examples, an example was described in which a compound represented by the above formula []-b was produced from a compound represented by the above formula []-a. The above formula [] -
The compound represented by b does not necessarily need to be produced by such a method, and the compound represented by the above formula []--
Of course, the compound represented by the above formula []-b can be produced from the compound represented by a, and the method of the present invention can be performed on this compound.

Reference example 1 (1) 3β-acetoxy-26,27-dinorcholest-5-en-24-one mp.; 148-151℃ nmr (CDCl ₃ , TMS, δ (ppm)); 5.36 (1H, m, C -6), 4.60 (1H, m, C
-3), 2.12 (3H, S, C-25), 2.02
(3H, S, C-3 acetyl), mass (m/e); 354 (M-60), 339 (M-60-15), 296,
255, 215, 213. ir (in CH ₂ Cl ₂ ); 1730 cm ^-1 elemental analysis (C ₂₇ H ₄₂ O ₃ ); Calculated value, C; 78.21, H; 10.21 Actual value, C; 78.12, H; 10.36 After heating and refluxing 2.730 g (6.58 mmol) and 220 mg of para-toluenesulfonic acid in acetic anhydride (50 ml) for 7 hours, about 20 ml of acetic anhydride was distilled off at normal pressure. After cooling the reaction mixture, it was poured into 0.5%-NaOH and extracted with ether-hexane (2:1). The organic layer was washed with water, dried (anhydrous sodium sulfate), and then separated and purified by silica gel column chromatography (carrier, SiO ₂ , 220 g). benzene-
From the ether (100:1) elution part, 3β,24-
Diacetoxy-26,27-dinorcholester-5,
23-Diene was obtained. Yield 2.162g (yield 72%) mp: 109-11℃ (reconsolidated from methanol), nmr (CDCl ₃ , TMS, δ); 5.40 (1H, m, C-6), 5.02 (1H, m, C
-23), 4.60 (1H, m, C-3), 2.14
(3H, S, C-240AC), 2.03 (3H, S, C
-30AC), 1.90 (C-25); mass (m/e); 396 (M-60), 354 (M-60-42), 283,
253, ir (in CH ₂ Cl ₂ ); 1750, 1735 cm ^-1 elemental analysis (C ₂₉ H ₄₄ O ₄ ); Calculated value, C; 76.27, H; 9.71 Actual value, C; 76.15, H; 9.77 Also, benzene - 313 mg of raw material was recovered from the ether (20:1) eluted portion.

(2) 2.162 g of the product of (1) above in an argon stream
(4.74 mmol) in 15 ml of diglyme, 170
Heated to ~185°C. Sodium chlorodifluoroacetate (10.84g) in diglyme solution (35g)
ml) was slowly added dropwise (about 15 minutes). After the dropwise addition was completed, the mixture was heated under reflux for 20 minutes. After cooling, the reaction mixture was poured into water (200 ml) and ether (100 ml), and after filtering out insoluble matter, the filtrate was extracted with hexane (300 ml), washed with water (200 ml x 2), and dried (anhydrous sodium sulfate). The product was separated and purified by silica gel column chromatography (SiO ₂ , 150 g), and the obtained crude crystals were recrystallized from methanol from the benzene eluted portion to obtain 819 mg of a compound represented by the following formula.

mp; 112-115℃ nmr (CDCl ₃ , TMS, δ (ppm)); 5.38 (1H, m, C-6), 4.60 (1H, m, C
-3), 2.05 (3H, S, C-240 acetyl),
2.02 (3H, S, C-30 acetyl), mass (m/e); 446 (M-60), 431 (M-60-15), 404 (M-60-42), 384 (M-60- 42-20), ir (in CH ₂ Cl ₂ ); 1760, 1730 cm ^-1 , elemental analysis (C ₃₀ H ₄₄ O ₄ F ₂ ); calculated value, C; 71.11, H; 8.75, F; 7.50 actual value, C; 71.26, H; 8.84, F; 7.50, (3) 394 mg (0.78 mmol) of the product from (2) above was dissolved in 15 ml of tetrahydrofuran (THF), and lithium hydroxide monohydrate (LiOH・H ₂ Add a water (10 ml)-methanol (4 ml) solution of O (140 mg),
After stirring at room temperature for 2.5 hours, water was added and extracted with ether. The ether extract was washed with water and dried (anhydrous NaSO ₄ ), concentrated and dried under reduced pressure, and the residue was dissolved in anhydrous methylene chloride (10 ml), pyridine (2.5 ml) and acetic anhydride (2.5 ml), and the solution was stirred at room temperature for 24 hours. . After concentration under reduced pressure, the residue was dissolved in ether, washed with dilute hydrochloric acid, 5% aqueous NaHCO ₃ solution and brine, and dried (anhydrous Na ₂ SO ₄ ). Separation by silica gel column chromatography. Purified (SiO ₂ , 200g). From the benzene elution part, 3β-acetoxy-24,24-difluoro-
I got 27-norcolester-5-en-25-on. Yield 43mg.

mp; 135-136.5°C (recrystallized from n-hexane), nmr ( _CDCl3 , TMS, δ); 5.32 (1H, m, C-6), 4.50 (1H, m, C
-3), 2.26 (3H, t, JHF=1Hz, C-
26), 1.96 (3H, S, C-30AC), mass (m/e); 404 (M-60), 389 (M-60-15), 296,
283, 255 ir (KBr); 1750, 1730, 1255, 1040 cm ^-1 Furthermore, from the benzene elution part, 3β-acetoxy-24-fluoro-27-norcholesta-5.23
34 mg of (E)-diene-25-one was obtained.

mp; 142-143℃ (recrystallized from methanol), nmr ( _CDCl3 , TMS); 5.62 (1H, dt, JHF=22Hz, JHH=8Hz,
C-23), 5.30 (1H, m, C-6), 4.52
(1H, m, C-3), 2.23 (3H, d, J=5
Hz, C-26), 1.96 (3H, S, C-30AC) mass (m/e); 384 (M-60), 369 (M-60-15), 283,
255, 253, 213 ir (KBr); 1730, 1705, 1640, 1255 cm ^-1 Elemental analysis (C ₂₈ H ₄₁ O ₃ F); Calculated value, C; 75.63, H; 9.30, F; 4.27 Actual value, C; 75.35, H; 9.52 Also, from the benzene-ether (30:1) elution part, 3β-acetoxy-24-fluoro-27-
234 mg of norcholester-5,23(Z)-dien-25-one was obtained.

mp; 169-170℃ (recrystallized from methanol), nmr ( _CDCl3 , TMS); 6.03 (1H, dt, JHF=34Hz, JHH=8Hz,
C-23), 5.35 (1H, m, C-6), 4.52
(1H, m, C-3), 2.27 (3H, d, J=3
Hz, C-26), 200 (3H, S, C-30AC) mass (m/e); 384 (M-60), 369 (M-60-15), 283,
256, 213 ir (KBr); 1730, 1695, 1650, 1240 cm ^-1 Elemental analysis (C ₂₈ H ₄₁ O ₃ F); Calculated value, C; 75.63, H; 9.30, F; 4.27 Actual value, C; 75.51, H; 9.37, F; 3.97 Example 1 In an argon stream, 70 mg of 3β-acetoxy-24,24-difluoro-27-norcholest-5-en-25-one obtained in (3) of Reference Example 1 above.
(0.15 mmol) was dissolved in 10 ml of anhydrous ether. Under ice-cooling and stirring, add excess CH ₃ MgI in ether (approx.
1.8 mmol) was added and stirred for 15 minutes. Dilute hydrochloric acid was added to the reaction mixture, and the mixture was extracted with ether. The ether extract was washed with water, dried (MgSO ₄ ), concentrated under reduced pressure,
The dried residue (containing 24,24-difluoro-25-hydroxycholest-5-ene) was dissolved in anhydrous methylene chloride (5 ml), pyridine (2 ml) and acetic anhydride (2 ml) for 20 hours at room temperature. Stirred. After concentration under reduced pressure, the residue was separated and purified by silica gel column chromatography (40 g of SiO ₂ ), and crude crystals were obtained from the portion eluted with benzene-ether (20:1). Yield 62mg. This was recrystallized from a mixed solvent of cyclohexane and n-hexane, and 3β-acetoxy-24,
24-difluorocholester-5-en-25-ol
Obtained 44mg.

mp; 163-164.5゜ nmr (CDCl ₃ , TMS); 5.38 (1H, m, C-6), 4.60 (1H, m, C
-3), 200 (3H, S, C-30 acetyl),
1.28 (6H, S, C-26, C-27) mass (m/e); 420 (M-60), 405 (M-60-15), 362,
312, 299, 255, 213 ir (KBr); 3400, 1730 cm ^-1 Elemental analysis (C ₂₉ H ₄₆ O ₃ F ₂ ); Calculated value, C; 72.46, H; 9.65, F; 7.90 Actual value, C; 72.50 , H; 9.70, F; 7.77 Example 2 30 mg of the product of Example 1 above was dissolved in 2 ml of carbon tetrachloride.
The mixture was heated to reflux, 16 mg of N-bromosuccinimide was added, and the mixture was heated to reflux for 25 minutes under an argon atmosphere. The mixture was cooled to room temperature, the precipitated imide crystals were removed by filtration, and the filtrate was concentrated and dried under reduced pressure.
The residue was dissolved in 1 to 2 ml of xylene and added dropwise to a solution of 1.5 ml of xylene and 0.5 ml of S-collidine that was being refluxed in an argon atmosphere, and the solution was further heated under reflux for 15 minutes. The crude product obtained by extraction with ethyl acetate and treatment in a conventional manner was dissolved in 5 ml of acetone, a catalytic amount of P-toluenesulfonic acid was added, and the mixture was stirred at room temperature under argon for 15 hours. The crude product obtained by extraction with ethyl acetate and conventional treatment was purified by preparative thin layer chromatography (developed three times with benzene:ethyl acetate = 15:1) to obtain 5 mg of 3β-
Acetoxy-24,24-difluorocholester-5,
7-diene-25-ol was obtained.

UV ( _λethanolnax ); 263, 272, 282, 292nm mass (m/e); 419 (M-(-C( _CH3 ) _2OH )), 418(M-
60), 403 (M-60-15), 313 (M-side chain),
253 (313-60) Reference Example 2 3 mg of 3β-acetoxy-24,24-difluorocholester-5,7-dien-25-ol
and tetrahydrofuran bubbled with argon 5
Dissolved in ml. This was bubbled with 5% argon.
5 ml of potassium hydroxide-methanol solution was added and left under argon for 15 hours. The product obtained by extraction with ethyl acetate and conventional treatment was dissolved in 40 ml of ethanol and 90 ml of benzene, and while bubbling argon gas, the product was exposed to ultraviolet light (Hanovia) while cooling on ice.
654A36, 200W) was irradiated for 2.5 hours. After refluxing for 1 hour in an argon stream, the solvent was removed under reduced pressure, and the resulting crude product was purified by preparative thin layer chromatography (developed 3 times with benzene:ethyl acetate = 5:1) to yield 0.76 g. This is further processed by high performance liquid chromatography (HPLC).
Purified by Zorbax SiL, CH ₂ Cl ₂ ), 0.2 mg of
24,24-difluoro-25-hydroxyvitamin
Got _D3 .

UV (ethanol); λ _nax = 228 nm λ _nio = 264 nm mass (m/e); 436 (M ⁺ ), 421 (M-15),
418 (M-18), 403 (M-15-18),
377, 271, 253, 136, 118

Claims (1)

[Claims] 1. The following formula [] [In the formula, R is a hydrogen atom or a hydroxyl group-protecting group. Also, 〓 represents a single bond or a double bond. ] 24,24-difluoro-25-hydroxycholestenes represented by.