A method for preparing a bovine somatotropin

Abstract

This application discloses a method for preparing Bosein. The method includes the following steps: mixing 1-C-(β-D-xylanosyl)-acetone, an alcohol solvent, an inorganic acid / base, and a reducing agent; reacting the mixture; filtering; desalting; decolorizing; and distilling to obtain Bosein with a purity of over 98%. The isomer ratio of the Bosein is in the range of (β, S) / (β, R) of 99:1 to 30:70. The isomer ratio is controllable, the operation is simple, and the obtained Bosein has a wide range of isomer ratios and good chemical purity.

Description

Technical Field

[0001] This application relates to a method for preparing bosine, which belongs to the field of organic synthesis. Background Technology

[0002] Pro-Xylane is a xylose derivative with anti-aging effects, also known as hydroxypropyltetrahydropyranotriol. Its chemical structure is 1-C-(β-D-xylanoside)-2-hydroxypropane. Studies have shown that hydroxypropyltetrahydropyranotriol can rebuild cell structure by promoting the formation of intercellular mucopolysaccharides, thereby promoting epidermal repair, improving skin elasticity, and combating skin aging.

[0003] The specific structure of Bosein (hydroxypropyltetrahydropyranotriol) is as follows:

[0004]

[0005] Hydroxypropyltetrahydropyranotriol, the main anti-wrinkle and anti-aging component of Pro-Xylane, exists in two diastereomers: 1-C-(β-D-xylanopyranoside)-2-(S)-hydroxypropane and 1-C-(β-D-xylanopyranoside)-2-(R)-hydroxypropane (hereinafter referred to as (β,S) / (β,R)). The ratio of these two isomers has a significant impact on the bioactivity of Pro-Xylane products. (Alexandre Cavezza et al., Synthesis of Pro-Xylane) TMA new biologically active C-glycoside inaqueous media, Bioorganic & Medicinal Chemistry Letters 19(2009)845-849, discloses a method for synthesizing C-glycosides in water. Specifically, it describes the synthesis of Bosein with a (β,S) / (β,R) ratio of 95 / 5 in a mixed solvent of isopropanol and acetic acid using sodium borohydride as a reducing agent, and the synthesis of Bosein with a (β,S) / (β,R) ratio of 50 / 50 using sodium borohydride as a reducing agent in water. Invention patent CN202311146214.5 discloses a dehydrogenase mutant and its application in the synthesis of S-Bosein, mainly using xylose and glucose as substrates and a dehydrogenase KPADH mutant as a catalyst to synthesize Bosein with a single (β,S) configuration. Invention patent CN202311146033.2 discloses the application and synthesis method of dehydrogenase in the synthesis of R-configuration Bosein, mainly using a combination of dehydrogenase SMADH2, glucose dehydrogenase GDH, coenzyme and glucose to synthesize a single (β, R) configuration of Bosein. Invention patent CN202211037235.9 discloses a method for producing colorless, odorless, and borate-free Bosein. This method separates water-soluble Bosein intermediates and products from the high-salt water phase after the reaction using a protecting group, and obtains a single (β, S) configuration of Bosein through chiral metal catalysis. Invention patent CN202111291311.4 discloses a method for synthesizing Bosein through acetal / ketal protection followed by reduction. This method involves reacting the intermediate with a dihydroxy protecting agent to form acetal or ketal, acetone or methoxybutanone groups to protect the hydroxyl group, followed by reduction and deprotection treatment to obtain a single (β, S) configuration of Bosein.

[0006] Existing processes for synthesizing Bosein include chemical synthesis and bio-enzyme catalysis. Both methods require different combinations of catalysts or enzymes to prepare Bosein with different (β, S) and (β, R) isomer ratios. The resulting Bosein configurations are uniform and the process is cumbersome, which is not conducive to the industrial scale-up of Bosein and results in high production line construction costs. Therefore, it is of great significance to develop a method that uses the same reducing agent and simply changes the concentration or amount of inorganic acid / base added to obtain Bosein with different (β, S) and (β, R) isomer ratios. Summary of the Invention

[0007] The purpose of this invention is to provide a method for controlling the configuration ratio of Bosein using inorganic acids / bases. Using the intermediate product 1-C-(β-D-xylanosyl)-acetone obtained from a condensation reaction as a raw material, a reduction reaction is carried out in an alcohol solvent by simply changing the concentration or amount of the added inorganic acid / base. This yields Bosein with a wide range of (β,S) / (β,R) isomer ratios, specifically 99:1 to 30:70. This method, using the same reducing agent and solvent, achieves wide-range control of the (β,S) / (β,R) isomer ratio by simply changing the concentration or amount of the inorganic acid / base. This simplifies the industrial scale-up process, reduces production line construction costs, and is beneficial for the industrial production of Bosein.

[0008] According to one aspect of this application, a method for preparing Bosein is provided, comprising the following steps:

[0009] 1-C-(β-D-xylanosyl)-acetone, alcohol solvent, inorganic acid / base and reducing agent are mixed, reacted, filtered, desalted, decolorized and distilled to obtain Bosein with a purity of more than 98%.

[0010] The ratio of the isomers of the bosine is in the range of (β, S) / (β, R) of 99:1 to 30:70.

[0011] Optionally, the following steps are included:

[0012] 1-C-(β-D-xylanosyl)-acetone was added to an alcohol solvent and stirred to dissolve. Then, an inorganic acid / base and a reducing agent were added sequentially, and the reaction was carried out. After the reaction was completed, the mixture was filtered, and the alcohol solvent was recovered by vacuum distillation. The distilled solid was dissolved in deionized water and desalted by passing it through an ion exchange column. The desalted aqueous solution was adsorbed and decolorized by activated carbon, and then distilled under reduced pressure to obtain the Bosein.

[0013] The alcohol solvent is selected from at least one of methanol, ethanol, n-propanol, isopropanol, glycerol, n-butanol, and sec-butanol;

[0014] The mass ratio of 1-C-(β-D-xylanosyl)-acetone to alcohol solvent is 1:1.1 to 1:8.

[0015] Optionally, the mass ratio of 1-C-(β-D-xylanosyl)-acetone to the alcohol solvent is any value from 1:1.1, 1:1.5, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8 or any range between the two.

[0016] The inorganic acid / base is selected from at least one of dilute hydrochloric acid, dilute sulfuric acid, dilute nitric acid, sodium hydroxide, sodium carbonate, sodium bicarbonate, potassium hydroxide, potassium carbonate, and potassium bicarbonate in an aqueous solution with a concentration of 3 to 10 wt%.

[0017] The mass ratio of the inorganic acid / base to 1-C-(β-D-xylanosyl)-acetone is 5:1 to 1:5.

[0018] Optionally, the mass ratio of the inorganic acid / base to 1-C-(β-D-xylanosyl)-acetone is any value from 5:1, 5:2, 5:3, 5:4, 5:5, 4:1, 4:2, 4:3, 3:1, 3:2 or any range between the two.

[0019] The reducing agent is selected from at least one of potassium borohydride, zinc borohydride, sodium borate, lithium aluminum hydride, sodium triacetoxyborohydride, and lithium triethylborohydride.

[0020] The molar ratio of 1-C-(β-D-xylanosyl)-acetone to the reducing agent is 1:1.3 to 1:1.8.

[0021] Optionally, the molar ratio of 1-C-(β-D-xylanosyl)-acetone to the reducing agent is any value among 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, and 1:1.8, or any range between the two.

[0022] The reaction temperature is 25–30°C;

[0023] Optionally, the temperature of the reaction is any value of 25°C, 26°C, 27°C, 28°C, 29°C, or 30°C, or a range between any two.

[0024] The reaction time is 1 to 3 hours.

[0025] Optionally, the reaction time is any value among 1h, 2h, and 3h, or a range between any two. The ratio of the isomers of the bosine is selected from one of 90:10, 80:20, 70:30, 60:40, 50:50, 40:60, and 30:70.

[0026] The ion exchange column is an anion and cation exchange resin column.

[0027] The mass ratio of activated carbon to the desalinated aqueous solution is 1:100 to 3:100.

[0028] The adsorption and decolorization temperature is 40–60°C;

[0029] The adsorption and decolorization time is 1 to 2.5 hours.

[0030] The beneficial effects that this application can produce include:

[0031] As a high-end raw material in anti-aging cosmetics, the ratio of its (β, S) / (β, R) isomers significantly affects its bioactivity and transdermal absorption. Existing synthesis processes for BOXIN include chemical synthesis and bio-enzymatic catalysis. Both methods require different combinations of catalysts or enzymes to prepare BOXIN with varying (β, S) and (β, R) isomer ratios. The resulting BOXIN configuration is limited by the type of catalyst or enzyme used, and the process is cumbersome, hindering industrial-scale production and resulting in high production line construction costs. Therefore, a solution is needed that allows for the production of BOXIN with different (β, S) and (β, R) isomer ratios by simply changing the reducing agent, solvent, or inorganic acid / base while maintaining the same process. The inorganic acids / bases used should be inexpensive and readily available, demonstrating significant industrial value. Attached Figure Description

[0032] Figure 1 The liquid chromatography spectrum of the (β, S) / (β, R) isomer obtained in Example 1 of the present invention with a ratio of 99 / 1 Bosein is obtained.

[0033] Figure 2 The liquid chromatography spectrum of the (β, S) / (β, R) isomer obtained in Example 2 of the present invention with a ratio of 80 / 20 Bosein.

[0034] Figure 3 The liquid chromatography spectrum of the (β, S) / (β, R) isomer obtained in Example 3 of the present invention with a ratio of 70 / 30 Bosein.

[0035] Figure 4 The liquid chromatography spectrum of the (β, S) / (β, R) isomer obtained in Example 4 of the present invention with a ratio of 60 / 40 Bosein.

[0036] Figure 5 The liquid chromatography spectrum of the (β, S) / (β, R) isomer obtained in Example 5 of the present invention with a ratio of 50 / 50 Bosein.

[0037] Figure 6 The liquid chromatography spectrum of the (β, S) / (β, R) isomer obtained in Example 6 of the present invention with a ratio of 40 / 60 Bosein.

[0038] Figure 7 The liquid chromatography spectrum of the (β, S) / (β, R) isomer obtained in Example 7 of the present invention with a ratio of 30 / 70 Bosein. Detailed Implementation

[0039] The present application is described in detail below with reference to the embodiments, but the present application is not limited to these embodiments.

[0040] Unless otherwise specified, all raw materials used in the embodiments of this application were purchased through commercial channels.

[0041] The intermediate product 1-C-(β-D-xylanosyl)-acetone was prepared according to the protocol described in L'Oréal's published literature Synthesis of Pro-Xylane™: A new biologically active C-glycoside in aqueous media.

[0042] The reducing agent and other components were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0043] The purity and (β, S) / (β, R) isomer determination of the Bosein product of this invention are performed according to existing technical methods.

[0044] Example 1

[0045]

[0046] 10 g of 1-C-(β-D-xylanosyl)-acetone was added to a 250 mL reaction flask and dissolved in 150 g of isopropanol. 15 g of sodium triacetoxyborohydride was added to the reaction flask, and the mixture was stirred at room temperature for 2 hours. After the reaction was stopped, the precipitate was removed by filtration, and the solvent was removed by vacuum distillation. The resulting deep yellow oily liquid was dissolved in 350 mL of deionized water and desalted by passing it through anion and cation exchange columns. 5 g of activated carbon was added to the aqueous solution, and the solution was decolorized in a 50 °C water bath for 2 hours. After decolorization, the solution was filtered, and the filtrate was concentrated by vacuum distillation. Ethyl acetate was added for recrystallization to obtain 6.5 g of white solid, which was Bosein. Liquid chromatography analysis showed that its purity was 99.9%, and the (β,S) / (β,R) isomer ratio was 99 / 1.

[0047] Example 2

[0048]

[0049] Take 31.25g of 32% dilute hydrochloric acid in a 250ml beaker, add 168.75g of deionized water to dilute it, and prepare 5% dilute hydrochloric acid.

[0050] 10 g of 1-C-(β-D-xylanosyl)-acetone was added to a 250 mL reaction flask and dissolved in 150 g of isopropanol. 3.64 g of 5% dilute hydrochloric acid was added to the dissolved reaction solution, and after uniform stirring, 3.6 g of sodium borohydride was slowly added to the reaction flask. The mixture was stirred at room temperature for 2 hours. After the reaction stopped, the precipitate was removed by filtration, and the solvent was removed by vacuum distillation. The resulting deep yellow oily liquid was dissolved in 350 mL of deionized water and desalted sequentially using anion and cation exchange columns. 5 g of activated carbon was added to the aqueous solution, and the solution was decolorized in a 50 °C water bath for 2 hours. After decolorization, the solution was filtered, and the filtrate was concentrated by vacuum distillation to obtain 5.6 g of a colorless, transparent, viscous liquid, which was Bosein. Liquid chromatography analysis showed that its purity was 99.8%, and the (β,S) / (β,R) isomer ratio was 80 / 20.

[0051] Example 3

[0052]

[0053] 10g of 1-C-(β-D-xylanosyl)-acetone was added to a 250mL reaction flask and dissolved in 150g of isopropanol. 8g of 5% dilute hydrochloric acid (prepared as described in Example 2) was added to the dissolved reaction solution. After uniform stirring, 3.6g of sodium borohydride was slowly added to the reaction flask, and the mixture was stirred at room temperature for 2 hours. After the reaction stopped, the precipitate was removed by filtration, and the solvent was removed by vacuum distillation. The resulting deep yellow oily liquid was dissolved in 350mL of deionized water and desalted sequentially using anion and cation exchange columns. 5g of activated carbon was added to the aqueous solution, and the solution was decolorized in a 50℃ water bath for 2 hours. After decolorization, the solution was filtered, and the filtrate was concentrated by vacuum distillation to obtain 6g of a colorless, transparent, viscous liquid, which was Bosein. Liquid chromatography analysis showed a purity of 99.7% and a (β,S) / (β,R) isomer ratio of 70 / 30.

[0054] Example 4

[0055]

[0056] 10g of 1-C-(β-D-xylanosyl)-acetone was added to a 250mL reaction flask and dissolved in 150g of isopropanol. 18g of 5% dilute hydrochloric acid (prepared as described in Example 2) was added to the dissolved reaction solution. After uniform stirring, 3.6g of sodium borohydride was slowly added to the reaction flask, and the mixture was stirred at room temperature for 2 hours. After the reaction stopped, the precipitate was removed by filtration, and the solvent was removed by vacuum distillation. The resulting deep yellow oily liquid was dissolved in 350mL of deionized water and desalted sequentially using anion and cation exchange columns. 5g of activated carbon was added to the aqueous solution, and the solution was decolorized in a 50℃ water bath for 2 hours. After decolorization, the solution was filtered, and the filtrate was concentrated by vacuum distillation to obtain 5.8g of a colorless, transparent, viscous liquid, which was Bosein. Liquid chromatography analysis showed a purity of 99.8% and a (β,S) / (β,R) isomer ratio of 60 / 40.

[0057] Example 5

[0058]

[0059] 10g of 1-C-(β-D-xylanosyl)-acetone was added to a 250mL reaction flask and dissolved in 150g of isopropanol. 36.5g of 5% dilute hydrochloric acid (prepared as described in Example 2) was added to the dissolved reaction solution. After uniform stirring, 3.6g of sodium borohydride was slowly added to the reaction flask, and the mixture was stirred at room temperature for 2 hours. After the reaction stopped, the precipitate was removed by filtration, and the solvent was removed by vacuum distillation. The resulting deep yellow oily liquid was dissolved in 350mL of deionized water and desalted sequentially using anion and cation exchange columns. 5g of activated carbon was added to the aqueous solution, and the solution was decolorized in a 50℃ water bath for 2 hours. After decolorization, the solution was filtered, and the filtrate was concentrated by vacuum distillation to obtain 6.1g of a colorless, transparent, viscous liquid, which was Bosein. Liquid chromatography analysis showed a purity of 99.8% and a (β,S) / (β,R) isomer ratio of 50 / 50.

[0060] Example 6

[0061]

[0062] 10 g of 1-C-(β-D-xylanosyl)-acetone was added to a 250 mL reaction flask and dissolved in 140 g of methanol. After stirring evenly, 3.6 g of sodium borohydride was slowly added to the reaction flask, and the mixture was stirred at room temperature for 2 hours. After the reaction stopped, the precipitate was removed by filtration, and the solvent was removed by vacuum distillation. The resulting deep yellow oily liquid was dissolved in 350 mL of deionized water and desalted by passing it through anion and cation exchange columns. 5 g of activated carbon was added to the aqueous solution and the solution was decolorized in a 50 °C water bath for 2 hours. After decolorization, the solution was filtered, and the filtrate was concentrated by vacuum distillation to obtain 5.9 g of colorless, transparent, viscous liquid, which was Bosein. Liquid chromatography analysis showed that its purity was 99.8%, and the (β,S) / (β,R) isomer ratio was 40 / 60.

[0063] Example 7

[0064]

[0065] Take 31.25g of 32% liquid alkali in a 250ml beaker, add 168.75g of deionized water to dilute, and prepare a 5% sodium hydroxide solution.

[0066] 10 g of 1-C-(β-D-xylanosyl)-acetone was added to a 250 mL reaction flask and dissolved in 150 g of isopropanol. 4.55 g of 5% sodium hydroxide solution was added to the dissolved reaction solution, and after uniform stirring, 3.6 g of sodium borohydride was slowly added to the reaction flask. The mixture was stirred at room temperature for 2 hours. After the reaction stopped, the precipitate was removed by filtration, and the solvent was removed by vacuum distillation. The resulting deep yellow oily liquid was dissolved in 350 mL of deionized water and desalted sequentially using anion and cation exchange columns. 5 g of activated carbon was added to the aqueous solution, and the solution was decolorized in a 50 °C water bath for 2 hours. After decolorization, the solution was filtered, and the filtrate was concentrated by vacuum distillation to obtain 6 g of a colorless, transparent, viscous liquid, which was Bosein. Liquid chromatography analysis showed that its purity was 99.8%, and the (β,S) / (β,R) isomer ratio was 30 / 70.

[0067] In summary, this application provides a method for controlling the configuration ratio of Bosein using inorganic acids / bases. Under the same process control, different (β, S) and (β, R) isomer ratios of Bosein can be obtained by simply changing the reducing agent, solvent, or inorganic acid / base. The inorganic acids / bases used are inexpensive and readily available, and have significant industrial value.

[0068] The above description is merely a few embodiments of this application and is not intended to limit this application in any way. Although this application discloses preferred embodiments as described above, it is not intended to limit this application. Any changes or modifications made by those skilled in the art without departing from the scope of the technical solution of this application using the disclosed technical content are equivalent to equivalent implementation cases and fall within the scope of the technical solution.

Claims

1. A method for preparing Bosein, characterized in that, Includes the following steps: 1-C-(β-D-xylanosyl)-acetone, alcohol solvent, inorganic acid / base and reducing agent are mixed, reacted, filtered, desalted, decolorized and distilled to obtain Bosein with a purity of more than 98%. The ratio of the isomers of the bosine is in the range of (β, S) / (β, R) from 99:1 to 30:

70.

2. The method according to claim 1, characterized in that, Includes the following steps: 1-C-(β-D-xylanosyl)-acetone was added to an alcohol solvent and stirred to dissolve. Then, an inorganic acid / base and a reducing agent were added sequentially, and the reaction was carried out. After the reaction was completed, the mixture was filtered, and the alcohol solvent was recovered by vacuum distillation. The distilled solid was dissolved in deionized water and desalted by passing it through an ion exchange column. The desalted aqueous solution was adsorbed and decolorized by activated carbon, and then distilled under reduced pressure to obtain the Bosein.

3. The method according to claim 1, characterized in that, The alcohol solvent is selected from at least one of methanol, ethanol, n-propanol, isopropanol, glycerol, n-butanol, and sec-butanol; The mass ratio of 1-C-(β-D-xylanosyl)-acetone to alcohol solvent is 1:1.1 to 1:

8.

4. The method according to claim 1, characterized in that, The inorganic acid / base is selected from at least one of dilute hydrochloric acid, dilute sulfuric acid, dilute nitric acid, sodium hydroxide, sodium carbonate, sodium bicarbonate, potassium hydroxide, potassium carbonate, and potassium bicarbonate in an aqueous solution with a concentration of 3 to 10 wt%. The mass ratio of the inorganic acid / base to 1-C-(β-D-xylanosyl)-acetone is 5:1 to 1:

5.

5. The method according to claim 1, characterized in that, The reducing agent is selected from at least one of potassium borohydride, zinc borohydride, sodium borate, lithium aluminum hydride, sodium triacetoxyborohydride, and lithium triethylborohydride. The molar ratio of 1-C-(β-D-xylanosyl)-acetone to the reducing agent is 1:1.3 to 1:1.

8.

6. The method according to claim 1, characterized in that, The reaction temperature is 25–30°C; The reaction time is 1 to 3 hours.

7. The method according to claim 1, characterized in that, The ratio of the isomers of the bosonicine is selected from one of 99:1, 90:10, 80:20, 70:30, 60:40, 50:50, 40:60, and 30:

70.

8. The method according to claim 2, characterized in that, The ion exchange column is an anion and cation exchange resin column.

9. The method according to claim 2, characterized in that, The mass ratio of activated carbon to the desalinated aqueous solution is 1:100 to 3:

100.

10. The method according to claim 2, characterized in that, The adsorption and decolorization temperature is 40–60°C; The adsorption and decolorization time is 1 to 2.5 hours.

Images