JPS6141360B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention provides 3β・25-dihydroxycholesto-
The present invention relates to a novel method for producing 5-ene, i.e., 25-hydroxycholesterol or its 3β-acetate. More specifically, the present invention relates to a novel method for producing 25-hydroxycholesterol or its 3β-acylate, which is an intermediate that can easily be converted into 25-hydroxycholecalciferol, which has recently attracted attention as active vitamin _D3 . Conventionally, methods for producing 25-hydroxycholesterol include the method of AIRyer et al. [see Journal of American Chemical Society (J.Am. ) and other methods (see J. Am. Chem. Soc, 72 4248 (1950)) are known. In the method of Layer et al., 25-hydroxycholesterol is obtained by reacting 3β-acetoxy-5-cholesten-25-one with methylmagnesium iodide, and in the latter method, the free hydroxy form of the above-mentioned sterol ester is used as the raw material. ,3
This method is almost the same as the former except that β-hydroxy-5-cholesten-25-one is used. Norcholesterol-25-one, which is the raw material for these methods, is obtained by oxidizing cholesterol, but the yield is poor and only norcholesterol-25-one must be separated and purified from various oxidized products. The major drawback is that it is difficult to obtain in large quantities because of the
Therefore, these methods cannot be said to be desirable as industrial methods for producing 25-hydroxycholesterol. An object of the present invention is to provide a method for industrially producing 25-hydroxycholesterol in good yield using simple operations starting from easily available raw materials. The method of the present invention consists of several reaction steps, which can be expressed by the following reaction formula. In the above reaction formula, Ac represents a CH ₃ CO- group,
R' is an aliphatic group, R'' is a hydrogen atom or a CH ₃ CO- group, R is an alkyl group or an aryl group, and X is a chlorine, bromine or iodine atom. As is clear at first glance, the entire process consists of 6 steps from the 1st step to the 6th step.
It consists of two steps, but the intermediate obtained up to the fourth step is 3α・6α・25-trihydroxy-
5β-cholestane, with three hydroxyl groups in free form. This means that all of the halogenating agents or ester residues used in the first to third steps are removed as a result. Further, the same can be said about the fifth step and the sixth step. Hereinafter, the method of the present invention will be explained in detail for each of the above-mentioned steps.Hyodeoxycholic acid 3α,6α-diacetate, which is the starting material of the present invention, is prepared by using ordinary hyodeoxycholic acid, which is available at low cost as a commercially available reagent or as an industrial product. It can be easily produced with good yield by an acetylation method, and is a highly valuable substance as a raw material for the industrial production of 25-hydroxycholesterol and its esters, as well as active vitamin _D3 . In the first step, hyodeoxycholic acid-3α・
The 6α-diacetate () and a halogenating agent (for example, thionyl chloride) are reacted in the presence of a base catalyst (for example, pyridine) if necessary. The reaction is carried out in a solvent inert to the halogenating agent under cooling or at room temperature. A particularly preferred solvent system is anhydrous benzene. The reaction was completed in 2 to 3 hours, and the reaction solution was concentrated under reduced pressure. The resulting residue was dissolved in benzene several times and concentrated under reduced pressure repeatedly to remove excess thionyl chloride and obtain crude hyodeoxycholic acid halide. Diacetate () is obtained. The obtained crude product () can be used as it is as a raw material for the second step. In the second step, the crude hyodeoxycholic acid halide diacetate () is reacted with diazomethane. The reaction can be carried out under cooling in a solvent inert to diazomethane. A particularly preferred solvent system is a benzene ether mixed solvent. The reaction is 1
The reaction was completed in ~2 hours, and after returning the reaction solution to room temperature, it was stirred while bubbling nitrogen for 1 to 2 hours to remove excess diazomethane, and then concentrated under reduced pressure to obtain crude diazoketone (). I can do it. The obtained crude product () can be used as it is as a raw material for the third step. In the third step, the diazoketone (2) is reacted with an aliphatic alcohol (eg, methanol, ethanol, etc.) in the presence of a catalytic amount of silver benzoate and an organic base (eg, triethylamine, pyridine, etc.). The reaction is carried out in a mixed solvent system of excess alcohol and an inert solvent that easily dissolves the raw material (eg, dichloromethane, chloroform, etc.). A particularly preferred solvent system is a mixed solvent of methanol and dichloromethane (1:1). The reaction is completed in a short time under stirring at room temperature. Isolation and purification of the product can be easily carried out by conventional methods such as solvent extraction, chromatography, recrystallization, etc. In the fourth step, the methyl-25 obtained in the above operation is
- Homohyodeoxycholate 3α・6α-diacetate () to methylmagnesium halide (X
=CI Br, I) is reacted. The reaction can be carried out in a solvent inert to the Grignard reagent, cooled or at room temperature. Particularly preferred solvent systems are ether or tetrahydrofuran. The reaction was terminated by stirring overnight, and the products 3α, 6α,
25-trihydroxy-5β-cholestane ()
can be easily obtained by conventional isolation and purification methods such as solvent extraction and recrystallization. In the fifth step, 3α, 6α, 25-trihydroxy-5β-cholestane () and an alkyl or allylsulfonyl chloride (e.g., p-toluenesulfonyl chloride, etc.) are mixed in a basic organic compound (e.g., pyridine, S-collidine, etc.). Make it react. The amount of alkyl or allylsulfonyl chloride used in the reaction is preferably 2 to 3 times the molar amount of the 3α·6α 25 -trihydroxy compound. The reaction proceeds by standing for a long time under cooling or at room temperature. Product 3α・6α・25-trihydroxy-
5β-Cholestane-3α・6α-disulfonate () can be obtained by isolating the reaction solution using a conventional method, such as adding ice water to the reaction solution, solvent extraction, drying with a desiccant, filtering and concentrating, and obtaining the crude product (). obtained as. The obtained crude product () can be used as it is as a raw material for the sixth step. In the final sixth step, the above product 3α・6α25
-trihydroxy-5β-cholestane-3α・6
α-Ditosylate () and a metal acetate (for example, potassium acetate) are reacted in a polar solvent (for example, acetic acid, acetic acid-free, dimethylformamide-H ₂ O, N-methylpyridine-H ₂ O, etc.). The reaction was completed in several hours under heating conditions, and 3 of the products were
β・25-dihydroxycholest-5-ene-3β
-Acetate ()-) can be isolated using the usual method, for example, by adding ice water to the reaction solution, and after solvent extraction,
By concentrating or directly subjecting the residue to a hydrolysis reaction, 3β25-dihydroxycholest-5-ene can be easily obtained by conventional purification methods such as chromatography and recrystallization.
In the hydrolysis reaction, the above product (-) is reacted with a metal hydroxide (eg, NaOH, KOH, etc.).
The reaction is carried out at room temperature or under cooling in a mixed solvent that can dissolve the lower alcohol and the compound (-).
A particularly preferred solvent system is a methanol benzene mixture. The reaction proceeds quantitatively in several hours, and the product can be easily isolated and purified by conventional methods such as solvent extraction, chromatography, recrystallization, etc. The invention will be explained in further detail in the following examples. Example 1 (a) 2231 mg (4.59 mmol) of hyodeoxycholic acid 3α·6α-diacetate () and 0.09 ml of pyridine were added to 46 ml of anhydrous benzene. The resulting solution was ice-cooled (approximately 5°C), and 2.07 ml of thionyl chloride was gradually added dropwise while stirring. At room temperature after dropping
Stirred for 2.5 hours. The reaction solution was concentrated under reduced pressure. An additional 25 ml of anhydrous benzene was added to the residue, and the mixture was repeatedly concentrated under reduced pressure to drive out thionyl chloride contained in the residue. The obtained solid product, crude hyodeoxycholic acid chloride 3α·6α-diacetate, was used as a raw material compound () without being purified. IR (KBr; cm ^-1 ); 1810, 1735, 1250. (b) The crude hyodeoxycholic acid chloride 3α・6α-diacetate () obtained in Example 1-(a) was dissolved in 27.5 ml of anhydrous benzene, and 0.5N-
The mixture was added dropwise to 60 ml of diazomethane ether solution while cooling and stirring. After the dropwise addition, the mixture was stirred for 1 hour while cooling (approximately 5° C.). The reaction solution was returned to room temperature and stirred for 1 hour while bubbling nitrogen to remove excess diazomethane. Further, the reaction solution was concentrated under reduced pressure to obtain a crude diazoketone body. The obtained diazoketone body was used as a raw material compound () without purification. IR (KBr, cm ^-1 ); 2950, 2090, 1735, 1640,
1240. (c) The crude diazoketone body () obtained in Example 1-(b) was mixed with 14.9 ml of methylene chloride and methanol.
A solution of 85.5 mg of silver benzoate dissolved in 0.86 ml of triethyleneamine was slowly added dropwise to the mixed solution of 19.3 ml under stirring at room temperature. After dripping, 30
The mixture was stirred at room temperature for several minutes. After confirming that the reaction was complete by thin layer analysis, water was added and the product was extracted with methylene chloride. The extract was washed sequentially with dilute hydrochloric acid, water, sodium bicarbonate solution and water. The extract was dried over anhydrous sodium sulfate, filtered, and concentrated. The concentrated residue was subjected to column chromatography packed with silica gel (solvent: benzene-ethyl acetate system) to obtain 1755 mg of methyl-25-homohyodexycholate-3α/6α-diacetate (). (Yield 74.4% from hyodeoxycholic acid 3α·6α-diacetate ()). Melting point; 93-95℃ (methanol) IR (KBr) cm ^-1 ); 2945, 2860, 1730, 1450,
1360, 1235, 1020. NMR (CDCI ₃ ) δ (ppm)); 0.63 (S, 3H,
C-18- _CH3 ) 0.96 (S, 3H, C-19-
CH ₃ ) 2.02 (S, 6H,

【formula】 )3.64(S, 3H,

[Formula] )4.70, 5.12 (mb, 2H, C-3 & C-6-
H) High Mars (M ⁺ −2×CH ₃ COON); 384,
3068 _{( C26H40O2 )} . Example 2 Methyl-25-homo-hyodeoxycholate-3
Dissolve 1.7gr (3.3ml) of α6α-diacetate () in 39.5ml of anhydrous tetrahydrofuran, and add 49.4ml of 1M methylmagnesium bromide-tetrahydrofuran (THF) solution to the ice-cooled solution with stirring.
ml was slowly added dropwise. After the dropwise addition, the reaction solution was returned to room temperature and further stirred overnight. After confirming the completion of the reaction by thin layer analysis, the reaction solution was cooled with ice, and a 20% ammonium chloride aqueous solution was added dropwise. The mixture was separated and extracted from an aqueous ammonium chloride solution with THF. THF separation and THF
The extracts were combined and concentrated under reduced pressure. Add water and ether to the residue, stir until completely dissolved, separate the ether solution, dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain 3α, 6α, 25-
A residue containing trihydroxy-5β-cholestane () was obtained. For purification, the residue was subjected to column chromatography packed with silica gel (solvent: benzene).
3α・6 by combining the fractions containing the product and concentrating under reduced pressure.
α・25-trihydroxy-5β-cholestane
1.197gr (86% yield) was obtained. Melting point: 192-193.5℃ (methanol-methylene chloride) IR (KBr; cm ^-1 ); 3375, 2940, 2860, 1463,
1370, 1030 NMR (CD ₃ OD; δ (ppm)); 0.68 (S, 3H,
C-18- _CH3 ) 0.92 (S, 3H, C-19- _CH3 )
1.16 (S, 6H, C-26 & 27-CH ₃ ) 3.52 (4.02
(bm, 2H, C-3 & 6-H) Mass; 420 (M〓), 402, 384, 376 High Mass (M〓- _H2O ); 402, 3515. Example 3 (a) 420 mg (1 mmol) of 3α・6α・25-trihydroxy-5β-cholestane was added to 2.5 mg of anhydrous pyridine.
Dissolved in ml. 572 mg (3 mmol) of p-toluenesulfonyl chloride was added to the obtained solution, and the solution was left standing at room temperature for 19 hours. After adding ice water to the reaction solution, dilute hydrochloric acid was added to make it acidic (PH 3 to 4), and extraction was performed three times with 10 ml of methylene chloride. After washing the extract with water, drying with anhydrous sodium sulfate, filtering, and concentrating, a crude solid product was obtained.
621 mg of α·6α·25-trihydroxy-5β-cholestane 3α·6α-ditosylate () was obtained. The obtained crude ditosylate compound () was used as an intermediate raw material compound () without being purified. IR (KBr; cm ^-1 ); 3440, 2950, 2870, 1600,
1450, 1354, 1170, 810. NMR (CDCI ₃ , δ (ppm)); 0.59 (S, 3H,
C-18- _CH3 ) 0.81 (S, 3H, C-19-
_CH3 ) 1.20 (S, 6H, C-26 & 27- _CH3 )
2.44 (S, 6H, C-3 & 6-tosyl group _CH3 )
7.20-7.90 (M, 8H, aroma take proton). (b) 364 mg (0.5 mmol) of 3α·6α·25-trihydroxy-5β-cholestane 3.6-ditosylate obtained in Example 3-(a) was dissolved in a mixed solvent of 3.6 ml of dimethylformamide and 0.4 ml of water. Add 546 mg (7 m) of potassium acetate to the resulting solution.
mol) was added and dissolved, and the mixture was heated and stirred at a bath value of 110 to 120°C for 5 hours. After the reaction was completed, the reaction solution was allowed to cool, poured into 10 ml of ice water, and the aqueous solution was extracted three times with 10 ml of ethyl acetate. After washing the obtained ethyl acetate extract with water,
It was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Obtained 3β・25-dihydroxycholesto
The residue containing 5-ene and its 3β-acetate was dissolved in 4 ml of a methanol-benzene (1:1) mixed solution. A 5% potassium hydroxide-methanol solution was added dropwise to the resulting mixed solution. After the dropwise addition, the mixture was stirred at room temperature for 2.5 hours, and after confirming the completion of the reaction by thin layer analysis, 6 ml of water and 6 ml of ether were added to the reaction solution to separate the layers. The aqueous solution was extracted twice with 6 ml of ether, and the ether separation and ether extract were combined and washed with dilute hydrochloric acid, saturated aqueous sodium bicarbonate solution, and saturated brine. The obtained ether solution was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a residue containing the product. The residue was separated on a silica gel thin layer plate (solvent benzene-ethyl acetate 5:1) and extracted with ethyl acetate to give 3β-25-dihydroxycholest-5-ene (25-hydroxycholesterol) 121/mg (yield 60 %) was obtained. Melting point: 177-80℃ (methanol) IR (KBr; cm ^-1 ); 3400, 2940, 1465, 1373,
1050 NMR (CDCI ₃ , δ (ppm)); 0.68 (S, 3H,
C-18- _CH3 ) 1.01 (S, 3H, C-19-
_CH3 ) 1.22 (S, 6H, C-26 & 27- _CH3 )
1.44 (S, 2H, C-3 & 25-OH) 3.52
(bm, 1H, C-3-H) 5.35 (bm, 1H, C
-6-H) High Mass (M ⁺・); 402, 3489
(C ₂₇ H ₄₆ O ₂ ), theoretical value 402, 3498 Example 4 (a) 210 mg (0.5 mmol) of 3α・6α・25-trihydroxy-5β-cholestane was added to anhydrous pyridine.
Dissolved in 1.25ml. 285.8 mg (1.5 mmol) of p-toluenesulfonyl chloride was added to the resulting solution.
was added, and after dissolution, the solution was allowed to stand at room temperature for 22 hours. After adding ice water to the reaction solution, dilute hydrochloric acid was added to make it acidic (PH 3-4), and extraction was performed four times with 5 ml of methylene chloride. After washing the extract with water, drying with anhydrous sodium sulfate, filtering and concentrating, crude 3α, 6α,
25-trihydroxy-5β-cholestane-3
356 mg of α·6α-ditosylate () was obtained. (b) Crude 3α・6α・25− obtained in Example 4-(a)
Trihydroxy-5β-cholestane-3α・6
3.56 mg of α-ditosylate was dissolved in a mixed solvent of 3.6 ml of dimethylformamide and 0.36 ml of water. Add 534mg (6.85ml) of potassium acetate to the resulting solution.
mol) was added and dissolved, and the mixture was heated and stirred at a bath temperature of 110 to 120°C for 5 hours. After the reaction was completed, the reaction solution was allowed to cool, poured into 10 ml of ice water, and the aqueous solution was extracted three times with 10 ml of ethyl acetate. After washing the obtained ethyl acetate extract with water,
It was dried over anhydrous sodium sulfate, filtered, and concentrated. Obtained 3β・25-dihydroxycholesto
The residue containing 5-ene and its 3β-acetate was dissolved in a mixed solution of 2.7 ml of anhydrous pyridine and 1.9 ml of acetic anhydride, and stirred at room temperature for 5 hours. After confirming that 3β·25-dihydroxycholest-5-ene had disappeared by thin layer analysis, the reaction solution was concentrated under reduced pressure. Add 5 ml of ether and 3 ml of water to the resulting residue, completely dissolve and separate the layers, wash the ether solution with dilute hydrochloric acid, saturated aqueous sodium bicarbonate solution and water, dry over anhydrous sodium sulfate, filter, and concentrate. A residue containing the product was obtained. The residue was separated on a silica gel thin layer plate (solvent: benzene:ethyl acetate = 9:1) and extracted with ethyl acetate to obtain 3β-25-dihydroxycholest-5-ene-3β-acetate (25-
hydroxycholesterol-3-acetate)
108.5 mg (49% yield was obtained. Melting point: 138-141°C (acetone) IR (KBr, cm ^-1 ): 3460, 2940, 1735, 1466,
1375, 1245, 1030 NMR (CDCI ₃ , δ (ppm)); 0.68 (S, 3H,
C-18- _CH3 ) 1.02 (S, 3H, C-19-
_CH3 ) 1.21 (S, 6H, C-26 & 27- _CH3 )
2.02 (S, 3H,

[Formula] ) 4.62 (bm, 1H, C-3-H) 5.38 (bm,
1H, C-6-H) Mass spectrum; 426 (M ⁺ -H ₂ O), 384
(M ⁺・−CH ₃ COOH)366(M ⁺・−
_CH3COOH − _H2O )

Claims (1)

[Claims] 1 Reacting 3α・6α・25-trihydroxy-5β-cholestane with an organic sulfonyl chloride represented by the following formula R-SO ₂ Cl (wherein R is an alkyl group or an aryl group). and the following formula (In the formula, the definition of R is the same as above) 3α·6α·25-trihydroxy-5β-cholestane-3α·6α-ditosylate is produced, and then this is reacted with a metal acetate in a polar solvent. 3β・25-dihydroxy-5α-cholest-5-ene-3β-
Method of manufacturing acetate. 2 Hyodeoxycholic acid-3α,6α-diacetate is reacted with a halogenating agent to produce hyodeoxycholic acid halide-3α,6α-diacetate, which is then reacted with dimethane to form the following formula: The diazoketone represented by is formed, which is then reacted with an aliphatic alcohol in the presence of silver benzoate and an organic base to form methyl-25-homohyodexycholate 3α·6α-diacetate, which is then reacted with 3α.6α.25-Trihydroxy-5β-cholestane is produced by reaction with methylmagnesium halide, which is then converted into the following formula: R-SO ₂ Cl where R is an alkyl group or an aryl group. It is reacted with an organic sulfonyl halide represented by the following formula, (In the formula, the definition of R is the same as above.) 3α・6α・25-trihydroxy-
Production of 3β·25-dihydroxy-5α-cholest-5-ene-3β-acetate, characterized in that 5β-cholestane-3α·6α-ditosylate is produced and then reacted with a metal acetate in a polar solvent. Law.