JP3128579B2 - Method for producing 2-hydroxyestrogen 3-glycoside - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a method for producing 2-hydroxyestrogen 3-glycoside.

[0002]

2. Description of the Related Art Catechol estrogens are known to be one of metabolites of estrogen hormones, and various studies have been made on the synthesis of catechol estrogens and derivatives thereof due to their interest in biological action. . Among them, regarding the synthesis of glycosides,
There are methods disclosed in the following documents. a) T. Nambara et al., "Chem. Pharm. Bull.", Vol. 18, pa.
ge 474 (1970), b) T. Nambara et al., "Chem. Pharm. Bull.", Vol. 18, pa.
ge 1191 (1970) and c) T. Ohkubo et al. "Chem. Pharm. Bull.", Vol. 36, pag.
e 419 (1988). In particular, 3-position glycoside of catechol estrogen 2-
With respect to the synthesis of hydroxyestrogen 3-glycoside, according to the above-mentioned references a) and b), starting from 3-deoxyestradiol acetate, through 14 steps, 2-hydroxyestradiol 3-glycol was obtained in an overall yield of 0.33%.
Glucuronide is synthesized.

[0003]

SUMMARY OF THE INVENTION The above-mentioned synthesis method according to the prior art is difficult to apply to mass production because the raw material is expensive, requires many steps for production, and has a low yield. Had a problem. Accordingly, it is an object of the present invention to provide a process for producing 2-hydroxyestrogens 3-glycosides suitable for production on a commercial scale.

[0004]

According to the present invention, the above object is achieved by a general formula

Embedded image (In the formula, R ₁ represents an acetoxyl group, R ₂ represents a hydrogen atom or an ethynyl group, or R ₂ together with R ₁ represents an oxygen atom, and R ₃ represents a hydrogen atom or an acetoxyl group. The glycosylation of the 2-acetyl estrogen form shown in

Embedded image Wherein R ₁ , R ₂ and R ₃ have the above-mentioned meaning, and R ₄ represents a glycosyl group in which a hydroxyl group, a carboxyl group and an amino group which are functional groups are protected.
Acetyl estrogen 3-glycoside form, followed by Bayer-Villiger oxidation to give the general formula

Embedded image (In the formula, R ₁ , R ₂ , R ₃ and R ₄ have the above-mentioned meaning), after converting to a 2-acetoxyestrogen 3-glycoside, a protective group is eliminated. General formula

Embedded image (Wherein R ₅ represents a glycosyl group, R ₆ represents a hydroxyl group, R ₇ represents a hydrogen atom or an ethynyl group,
Or R ₇ represents an oxygen atom together with R _6, and R ₈ represents a hydrogen atom or a hydroxyl group). Is achieved.

[0005] In the method of the present invention, the 2-acetyl estrogen compound used as a starting material converts estrogen into 3-acetate, and the 3-acetate compound is converted to a fleece (Frie
s) It can be easily manufactured by rearrangement. By employing the method of the present invention, the total yield from estrogen to 2-hydroxyestrogen 3-glycoside can be increased to about 30%.

Next, starting from estradiol, 2-
This will be described in detail in connection with the production of hydroxyestradiol 3-glucopyranoside. First, by reacting estradiol and acetic anhydride in pyridine at room temperature overnight, estradiol diacetate is produced quantitatively. The estradiol diacetate is reacted with aluminum chloride and acetyl chloride in nitrobenzene at 80 ° C. for 1 hour to obtain 2-acetylestradiol 17-acetate in a yield of 85%. Then, the 2-acetylestradiol 17-
When acetate is reacted with acetobromoglucose in toluene in the presence of cadmium carbonate, two anomeric isomers, namely 2,3,4,6-tetra-O-acetyl-1-O- (2- Acetyl-17β-acetoxy-1,3,5 (10) -estraditriene-
3-yl) -β-D-glucopyranoside (yield: 39%) and 2,
3,4,6-tetra-O-acetyl-1-O- (2-acetyl-17β-acetoxy-1,3,5 (10) -estraditrien-3-yl) -α-D-glucopyranoside (yield : 48%). Separate each product and oxidize it with Bayer Villiga
If the acetyl group is eliminated under alkaline conditions after conversion to the acetoxy form, the desired 2-hydroxyestradiol 3-β-D-glucopyranoside can be obtained in a yield of 83%, or
2-Hydroxyestradiol 3-α-D-glucopyranoside is obtained with a yield of 70%. The overall yield of these desired products from estradiol is 28% for the former and 2% for the latter.
9%.

[0007] The above-mentioned manufacturing process can be represented by the following reaction formula.

Embedded image

Embedded image

[0008]

Next, the present invention will be described in more detail and concretely with reference examples and examples.

Reference Example 1 (Production of estradiol diacetate) Estradiol (30 g, 110 mmol) was mixed with pyridine (300 ml).
(3.71 mol) and 105 ml (1.08 mol) of acetic anhydride were added, and the mixture was stirred at room temperature overnight. The reaction mixture was poured into 1500 g of ice, and the precipitated solid was collected by filtration and sufficiently washed with water. The obtained white solid was dried to obtain 39.3 g (yield: 100%) of a desired estradiol diacetate. ¹ H-NMR spectrum (DMSO-d ₆ ) δ ppm: 0.78 (3H, s, 18-CH ₃ ), 1.1-2.9 (15H, m, CH and CH ₂ ), 2.01 (3H, s, C ₁₇ -OCOCH ₃ ), 2.23 (3H, s, C ₃ -OCOCH ₃ ), 4.62 (1H, t like, C ₁₇ -H), 6.83 (2H, m, C ₂ -H and C ₄ -H), 7.28 (1H, d, J = 10Hz, C ₁ -H).

Reference Example 2 (2-acetylestradiol 1
Production of 7-acetate) 25.0 g of estradiol diacetate obtained in Reference Example 1
(70.1 mmol) was dissolved by adding 250 ml of nitrobenzene. 18.6 g (139 mmol) of aluminum chloride was added to this solution.
And 2.0 ml (28 mmol) of acetyl chloride at 80 ° C.
Stir for 1 hour. After the reaction mixture was ice-cooled, the cold solution was poured into 1000 g of ice, then 100 ml of concentrated hydrochloric acid was added, and the mixture was extracted four times with chloroform. Merge organic layers, 1
Washed with N hydrochloric acid. Next, after washing with a saturated aqueous solution of sodium hydrogencarbonate and a saturated saline solution, the solution was concentrated with a vacuum evaporator to obtain 25.3 g of a crude product.
To this was added 150 ml of methanol to obtain 17.7 g (yield: 70.6%) of desired 2-acetylestradiol 17-acetate as a methanol-insoluble portion. The methanol wash was concentrated and subjected to silica gel column chromatography (eluent, chloroform: hexane = 2: 1) to give 3.48 g of the desired compound. Total yield was 85%. ¹ H-NMR spectrum (CDCl ₃ ) δ ppm: 0.84 (3H, s, 18-CH ₃ ), 1.2-3.0 (15H, m, CH and CH ₂ ), 2.07 (3H, s, C ₁₇ -OCOCH ₃ ) , 2.59 (3H, s, C ₂ -COCH ₃ ), 4.70 (1H, t like, C ₁₇ -H), 6.67 (1H, s, C ₄ -H), 7.59 (1H, s, C ₁ -H) , 12.03 (1H, s, C ₃ -OH).

Example 1 a) 2,3,4,6-Tetra-O-acetyl-1-O- (2-acetyl-17β-
Acetoxy-1,3,5 (10) -estraditrien-3-yl) -β-D
-Glucopyranoside and 2,3,4,6-tetra-O-acetyl-1-
Production of O- (2-acetyl-17β-acetoxy-1,3,5 (10) -estraditrien-3-yl) -α-D-glucopyranoside] 2-acetylestradiol 17-acetate 891 obtained in Reference Example 2 mg (2.50 mmol), cadmium carbonate 1.72 g (1
0.0 mmol) and 50 ml of toluene were weighed into a flask, and heated in an oil bath to distill off 15 ml of toluene. Then acetobromoglucose 3.08 g (7.50 mmol)
Was added dropwise over 2 hours, and toluene was constantly distilled off in order to azeotropically remove water produced at this time from the reaction system. To maintain the liquid volume, toluene was appropriately added. After the completion of the dropwise addition, the mixture was continuously distilled off for another 2 hours while adding only toluene. at this point
When TLC analysis was carried out, the raw material remained, and after adding 0.86 g (5.00 mmol) of cadmium carbonate, a toluene solution of 1.54 g (3.75 mmol) of acetobromoglucose 30 was added.
ml was added dropwise over 1 hour. At this time, as in the case described above, toluene was continuously distilled off in order to azeotropically remove generated water from the reaction system. 1.5 hours after dropping
At the same time, the mixture was continuously distilled while only toluene was added dropwise. Next, the reaction mixture was filtered through celite, and the filtrate was concentrated under reduced pressure to obtain a crude product. This was separated and purified by silica gel column chromatography (elution solvent, hexane: ethyl acetate = 2: 1) to give the desired 2,3,4,6-
Tetra-O-acetyl-1-O- (2-acetyl-17β-acetoxy-
662 mg of 1,3,5 (10) -estraditrien-3-yl) -β-D-glucopyranoside (yield: 38.5%), and 2,3,4,6-tetra-O
-Acetyl-1-O- (2-acetyl-17β-acetoxy-1,3,5 (1
0) -Estratrien-3-yl) -α-D-glucopyranoside
1.23 g (purity 67%, net weight 824 mg, yield: 47.9%) was obtained. Analytical data: 2,3,4,6-tetra-O-acetyl-1-O- (2-acetyl-17β-acetoxy-1,3,5 (10) -estraditriene-3-
Yl) -β-D-glucopyranoside ¹ H-NMR spectrum (CDCl ₃ ) δ ppm: 0.82 (3H, s, 18-CH ₃ ), 1.2-3.0 (30H, m, estrogen CH and CH ₂ and O
COCH ₃ x 5), 2.52 (3H, s, C ₂ -COCH ₃ ), 3.8-4.3 (3H, m, pyranose C ₅ -H and C ₆ -H ₂ ), 4.68 (1H, t like, 17α-H ), 5.1-5.4 (4H, m, pyranose C ₁ -H, C ₂ -H, C ₃ -H and C
₄ -H), 6.76 (1H, s, Ar C ₄ -H), 7.61 (1H, s, Ar C ₁ -H). Silica gel TLC: Rf = 0.12 (Hexane: Ethyl acetate = 2: 1). 2, 3,4,6-tetra-O-acetyl-1-O- (2-acetyl-17β-acetoxy-1,3,5 (10) -estraditrien-3-yl) -α-D-
Glucopyranoside ¹ H-NMR spectrum (CDCl ₃ ) δ ppm: 0.83 (s, 18-CH ₃ ), 1.2-3.0 (m, estrogen CH and CH ₂ and OCOCH ₃
x 5), 2.73 (s, C ₂ -COCH ₃ ), 3.8-4.4 (m, pyranose C ₅ -H and C ₆ -H ₂ ), 4.68 (t like, 17α-H), 5.0-5.8 (m, pyranose _{C 1 -H, C 2 -H,} C 3 -H and C 4 -
H), 6.96 (s, Ar C ₄ -H), 7.68 (s, Ar C ₁ -H). Silica gel TLC: Rf = 0.18 (Hexane: Ethyl acetate = 2: 1).

B) 2,3,4,6-tetra-O-acetyl-1-O- (2,17
β-diacetoxy-1,3,5 (10) -estraditrien-3-yl)
Production of -β-D-glucopyranoside 2,3,4,6-tetra-O-acetyl-1-O- (2-
Acetyl-17β-acetoxy-1,3,5 (10) -estraditrien-3-yl) -β-D-glucopyranoside 395 mg (0.575 mmo
l), 298 mg (1.73 mmol) of m-chloroperbenzoic acid and 245 mg (1.73 mmol) of disodium hydrogen phosphate are weighed in a flask, and then 8 ml of chloroform is added thereto.
Stir for 17 hours. After filtering off the insolubles in the reaction mixture, the chloroform layer was poured into ice-cooled diethyl ether, and washed sequentially with a 5% aqueous sodium carbonate solution, water, and then saturated brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure using an evaporator to obtain the desired 2,3,4,6-tetra-O-acetyl-1-O- (2,17β
-Diacetoxy-1,3,5 (10) -estraditrien-3-yl)-
440 mg of β-D-glucopyranoside was obtained as a crude product.
Although this material contains minor impurities, it is near pure in NMR spectrum and can therefore be used in subsequent reactions without further purification. ¹ H-NMR spectrum (CDCl ₃ ) δ ppm: 0.80 (3H, s, 18-CH ₃ ), 1.1-3.0 (30H, m, estrogen CH and CH ₂ and O
COCH ₃ x 5), 2.25 (3H, s, C ₂ -OCOCH ₃ ), 3.8-4.4 (3H, m, pyranose C ₅ -H and C ₆ -H ₂ ), 4.68 (1H, t like, 17α-H ), 5.0-5.4 (4H, m, pyranose C ₁ -H, C ₂ -H, C ₃ -H and
C ₄ -H), 6.71 (1H, s, Ar C ₄ -H), 6.93 (1H, s, Ar C ₁ -H). Silica gel TLC: Rf = 0.56 (chloroform: methanol = 20: 1).

C) 2-hydroxyestradiol 3-β-D
Preparation of -glucopyranoside 2,3,4,6-tetra-O-acetyl-1-O- (2,
To 400 mg of the crude product of 17β-diacetoxy-1,3,5 (10) -estraditrien-3-yl) -β-D-glucopyranoside was added 10 ml of methanol and 5.0 ml of 1N-sodium hydroxide for 1.5 hours at room temperature. Stirred. Next, after removing methanol with an evaporator, 15 ml of water and 150 μl of acetic acid were added. After the precipitated solid was collected by filtration, it was subjected to silica gel column chromatography (elution solvent, chloroform:
Methanol = 5: 1)
2-hydroxyestradiol 3-β-D-glucopyranoside 196 mg (Yield: 2,3,4,6-tetra-O-acetyl-1-O- (2-
Acetyl-17β-acetoxy-1,3,5 (10) -estraditrien-3-yl) -β-D-glucopyranoside (83.1%). ¹ H-NMR spectrum (CD ₃ OD) δ ppm: 0.76 (3H, s, 18-CH ₃ ), 1.0-2.8 (15H, m, estrogen CH and CH ₂ ), 3.3-4.0 (7H, m, estrogen 17α -H and pyranose C ₂ -H, C ₃ -H, C ₄ -H, C ₅ -H and C ₆ -H ₂ ), 4.64-4.72 (1H, m, pyranose C ₁ -H), 6.77 (1H, _{. s, Ar C 4 -H)} , 6.86 (1H, s, Ar C 1 -H) 13 C-NMR spectrum _{(DMSO-d 6) δ ppm} : 11.16, 22.76,
26.01, 27.04, 28.50, 29.91, 36.63, 38.41, 42.75, 4
3.67, 49.58, 60.79, 69.84, 73.36, 75.91, 77.15, 79.
97, 103.05, 112.64, 117.68, 126.73, 134.80, 143.0
9,144.71.Silica gel TLC: Rf = 0.23 (chloroform: methanol = 5: 1).

Example 2 a) 2,3,4,6-Tetra-O-acetyl-1-O- (2,17β-diacetoxy-1,3,5 (10) -estraditrien-3-yl)- Production of α-D-glucopyranoside 2,3,4,6-tetra-O-acetyl-1-O- obtained in Example 1-a)
790 mg of (2-acetyl-17β-acetoxy-1,3,5 (10) -estraditrien-3-yl) -α-D-glucopyranoside (purity 67
%, Net weight 529mg, 0.771mmol), m-chloroperbenzoic acid
596 mg (3.45 mmol) and disodium hydrogen phosphate 490 mg
(3.45 mmol) was weighed into a flask and then chloroform
16 ml was added and the mixture was stirred at room temperature for 22 hours. After filtering off insolubles in the reaction mixture, the chloroform layer was poured into ice-cooled diethyl ether, and washed sequentially with a 5% aqueous sodium carbonate solution, water, and then saturated brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure using an evaporator to give the desired 2,3,4,6-tetra-O-acetyl-1-O- (2,17β-diacetoxy-1, 3,5 (10)
Thus, 863 mg of -estraditrien-3-yl) -α-D-glucopyranoside was obtained as a crude product. This material contains impurities, but can be used as is in subsequent reactions. ¹ H-NMR spectrum (CDCl ₃ ) δ ppm: 0.82 (s, 18-CH ₃ ), 1.1-2.9 (m, estrogen CH and CH ₂ and OCOCH ₃
x 5), 2.37 (s, C ₂ -OCOCH ₃ ), 3.7-4.4 (m, pyranose C ₅ -H and C ₆ -H ₂ ), 4.69 (t like, 17α-H), 4.9-5.8 (m, pyranose _{C 1 -H, C 2 -H,} C 3 -H and C 4 -
H), 6.91 (s, Ar C ₄ -H), 6.98 (s, Ar C ₁ -H). Silica gel TLC: Rf = 0.44 (chloroform: methanol = 50: 1).

B) 2-hydroxyestradiol 3-α-D
Preparation of 2-glucopyranoside 2,3,4,6-tetra-O-acetyl-1-O- (2,
20 ml of methanol and 10 ml of 1N-sodium hydroxide were added to 850 mg of the crude product of 17β-diacetoxy-1,3,5 (10) -estraditrien-3-yl) -α-D-glucopyranoside,
The mixture was stirred at room temperature for 1 hour. Next, after removing methanol with an evaporator, 50 ml of water and 366 μl of acetic acid were added. The precipitated solid was collected by filtration and crystallized from methanol to give 240 mg of the desired 2-hydroxyestradiol 3-α-D-glucopyranoside (yield: 2,2).
70.4% from 3,4,6-tetra-O-acetyl-1-O- (2-acetyl-17β-acetoxy-1,3,5 (10) -estraditrien-3-yl) -α-D-glucopyranoside ). ¹ H-NMR spectrum (DMSO-d ₆ : CD ₃ OD = 3: 1) δ ppm
: 0.71 (3H, s, 18-CH ₃ ), 1.0-2.8 (15H, m, estrogen CH and CH ₂ ), 3.3-3.9 (7H, m, estrogen 17α-H and pyranose C ₂ -H, C _3- H, C ₄ -H, C ₅ -H and C ₆ -H ₂ ), 5.14 (1H, d, J = 3 Hz, pyranose C ₁ -H), 6.73 (1H, s, Ar C ₄ -H), 6.88 (1H, s, Ar C ₁ -H). ¹³ C-NMR spectrum (DMSO-d ₆ ) δ ppm: 11.16, 22.70,
26.01, 27.04, 28.39, 29.91, 36.68, 38.36,42.75, 4
3.67, 49.58, 60.68, 69.95, 71.95, 73.14, 73.58, 80.
02, 100.50, 112.53, 118.01, 126.68, 134.80, 143.0
4,145.14. Silica gel TLC: Rf = 0.23 (chloroform: methanol = 5: 1).

[0016]

According to the method of the present invention, 2-hydroxyestrogens 3-glycosides can be produced in three steps starting from 2-acetylestrogens, the yield is good, and the method is suitable for mass production. is there. In addition, when starting from estrogen, there are 5 steps, and the total yield is about 30%.
(approximately 60% when α-form and β-form are combined). The method of the present invention is remarkably superior to the conventional method of synthesizing 2-hydroxyestradiol 3-glucuronide in a total yield of 0.33% through 14 steps starting from 3-deoxyestradiol acetate.

──────────────────────────────────────────────────続 き Continuation of front page (72) Kunio Yagi, Inventor 2-21 Nishisato-cho, Meito-ku, Nagoya-shi, Aichi (58) Field surveyed (Int. Cl. ⁷ , DB name) C07J 75/00 C07J 17/00 CA (STN)

Claims (1)

(57) [Claims] 1. A compound of the general formula (In the formula, R ₁ represents an acetoxyl group, R ₂ represents a hydrogen atom or an ethynyl group, or R ₂ together with R ₁ represents an oxygen atom, and R ₃ represents a hydrogen atom or an acetoxyl group. The glycosylation of the 2-acetyl estrogen compound represented by Wherein R ₁ , R ₂ and R ₃ have the above-mentioned meaning, and R ₄ represents a glycosyl group in which a hydroxyl group, a carboxyl group and an amino group which are functional groups are protected.
Acetyl estrogen 3-glycoside form, followed by Bayer-Villiger oxidation, yields the general formula: (In the formula, R ₁ , R ₂ , R ₃ and R ₄ have the above-mentioned meaning), after converting to a 2-acetoxyestrogen 3-glycoside, a protective group is eliminated. General formula (Wherein R ₅ represents a glycosyl group, R ₆ represents a hydroxyl group, R ₇ represents a hydrogen atom or an ethynyl group,
Or R ₇ together with R ₆ represents an oxygen atom, and R ₈ represents a hydrogen atom or a hydroxyl group).