Method for obtaining anhydroicaritin from icariin by adopting naringinase

Abstract

The invention relates to a method for converting icariin to anhydroicaritin by biological enzyme reaction so as to improve the biological activity. Under the action of naringinase, the reaction is performed in an ethanol water solution with the concentration of 30%-70%. The reaction temperature is 40-70 DEG C, and the reaction time is 1-30h. A glycosyl group on a hydroxyl group of icariin is cut off, and then icariin is converted to anhydroicaritin with higher pharmacological activity. By adopting the method provided by the invention, the processability of icariin is improved, and meanwhile, the actual application range can be expanded.

Description

technical field

[0001] The invention relates to a method for converting icariin into icariin, in particular to a method for obtaining icariin by naringinase reaction. Background technique

[0002] Icariin (Icratin) molecular formula is C 33 h 40 o 15 , the chemical structure is as shown. Pale yellow needle crystal.

[0003]

[0004] Icariin is derived from natural products. The natural active substances icariin and icariin have great development value. At present, icariin and icariin are mostly used in the clinical treatment of cardiovascular diseases.

[0005] Icariin can significantly inhibit myocardial contractility and reduce the rate of increase of left ventricular pressure, indicating that it can reduce myocardial oxygen consumption, while reducing peripheral resistance, reducing cardiac afterload, and has therapeutic effects on cardiovascular diseases such as myocardial ischemia and arrhythmia. Improvement effect.

[0006] Icariin has obvious effects on t...

Examples

example 1

[0021] Example 1: Into a 1000mL conical flask, add 10mg of icariin standard substance and 400ml of 30% ethanol aqueous solution, adjust the pH value to 4.0 with 1M NaOH solution, and the system reaches a certain temperature of 50°C, then add 0.5g of naringinase (standard activity 475AGUPg), and finally, at 50°C and 200 rpm, the reaction was stirred for 30 hours. After the reaction, extract with the same volume of ethyl acetate to remove sugar and enzyme protein impurities. The extract was developed on a G silica gel plate (100mm×25mm), chloroform:methanol=8:2 as the mobile phase, and was developed under 245nm ultraviolet light. Observed below and carried out qualitative analysis with TLC, the results are as follows figure 1 shown in (1.R f淫羊藿苷标准品 =0.50,

[0022] 2. R f反应产物 =0.8), the reaction product R f The value 0.8 is much larger than the standard R f A value of 0.5 shows that the polarity of icariin less than icariin has been produced by the enzymatic reaction, and th...

example 2

[0025] Example 2: In a 1000ml Erlenmeyer flask, add 10mg of icariin standard substance and 400ml of 40% ethanol aqueous solution, adjust the pH value to 4.0 with 1M NaOH solution, and the system reaches a certain temperature of 50°C, then add 0.5g of naringinase (standard activity 475AGUPg), and finally, at 50°C and 200 rpm, the reaction was stirred for 30 hours. After the reaction, extract with the same volume of ethyl acetate to remove sugar and enzyme protein impurities. The extract is developed on a G silica gel plate (100mm×25mm), chloroform:methanol:water=7.5:2.5:0.25 as the mobile phase, Observe under 245nm ultraviolet light and carry out qualitative analysis with TLC, the result is as follows figure 2 shown (R f淫羊藿苷标准品 =0.42, 2.R f反应产物 ==0.8) Reaction product R f The value of 0.8 is much larger than the standard R f A value of 0.42 indicates that icariin, which is less polar than icariin, is produced by the enzymatic reaction, and the reaction mechanism is the sam...

example 3

[0026] Example 3: In a 1000ml Erlenmeyer flask, add 10mg of icariin standard substance and 400ml of 30% ethanol aqueous solution, adjust the pH value to 6.0 with 1M NaOH solution, and the system reaches a certain temperature of 60°C, then add 0.5g of naringinase (standard activity 475AGUPg), and finally, at 60°C and 200 rpm, the reaction was stirred for 30 hours. After the reaction, extract with the same volume of ethyl acetate to remove sugar and enzyme protein impurities. The extract is developed on a G silica gel plate (100mm×25mm), chloroform:methanol:water=7.5:2.5:0.25 as the mobile phase, Observe under 245nm ultraviolet light and carry out qualitative analysis with TLC, the result is as follows figure 2 shown (R f淫羊藿苷标准品 =0.42, 2.R f反应产物 = 0.8) Reaction product R f The value of 0.8 is much larger than the standard R f A value of 0.42 indicates that icariin, which is less polar than icariin, is produced by the enzymatic reaction, and the reaction mechanism is the sam...

technical field

[0001] The invention relates to a method for converting icariin into icariin, in particular to a method for obtaining icariin by naringinase reaction. Background technique

[0002] Icariin (Icratin) molecular formula is C 33 h 40 o 15 , the chemical structure is as shown. Pale yellow needle crystal.

[0003]

[0004] Icariin is derived from natural products. The natural active substances icariin and icariin have great development value. At present, icariin and icariin are mostly used in the clinical treatment of cardiovascular diseases.

[0005] Icariin can significantly inhibit myocardial contractility and reduce the rate of increase of left ventricular pressure, indicating that it can reduce myocardial oxygen consumption, while reducing peripheral resistance, reducing cardiac afterload, and has therapeutic effects on cardiovascular diseases such as myocardial ischemia and arrhythmia. Improvement effect.

[0006] Icariin has obvious effects on t...

Examples

example 1

[0021] Example 1: Into a 1000mL conical flask, add 10mg of icariin standard substance and 400ml of 30% ethanol aqueous solution, adjust the pH value to 4.0 with 1M NaOH solution, and the system reaches a certain temperature of 50°C, then add 0.5g of naringinase (standard activity 475AGUPg), and finally, at 50°C and 200 rpm, the reaction was stirred for 30 hours. After the reaction, extract with the same volume of ethyl acetate to remove sugar and enzyme protein impurities. The extract was developed on a G silica gel plate (100mm×25mm), chloroform:methanol=8:2 as the mobile phase, and was developed under 245nm ultraviolet light. Observed below and carried out qualitative analysis with TLC, the results are as follows figure 1 shown in (1.R f淫羊藿苷标准品 =0.50,

[0022] 2. R f反应产物 =0.8), the reaction product R f The value 0.8 is much larger than the standard R f A value of 0.5 shows that the polarity of icariin less than icariin has been produced by the enzymatic reaction, and th...

example 2

[0025] Example 2: In a 1000ml Erlenmeyer flask, add 10mg of icariin standard substance and 400ml of 40% ethanol aqueous solution, adjust the pH value to 4.0 with 1M NaOH solution, and the system reaches a certain temperature of 50°C, then add 0.5g of naringinase (standard activity 475AGUPg), and finally, at 50°C and 200 rpm, the reaction was stirred for 30 hours. After the reaction, extract with the same volume of ethyl acetate to remove sugar and enzyme protein impurities. The extract is developed on a G silica gel plate (100mm×25mm), chloroform:methanol:water=7.5:2.5:0.25 as the mobile phase, Observe under 245nm ultraviolet light and carry out qualitative analysis with TLC, the result is as follows figure 2 shown (R f淫羊藿苷标准品 =0.42, 2.R f反应产物 ==0.8) Reaction product R f The value of 0.8 is much larger than the standard R f A value of 0.42 indicates that icariin, which is less polar than icariin, is produced by the enzymatic reaction, and the reaction mechanism is the sam...

example 3

[0026] Example 3: In a 1000ml Erlenmeyer flask, add 10mg of icariin standard substance and 400ml of 30% ethanol aqueous solution, adjust the pH value to 6.0 with 1M NaOH solution, and the system reaches a certain temperature of 60°C, then add 0.5g of naringinase (standard activity 475AGUPg), and finally, at 60°C and 200 rpm, the reaction was stirred for 30 hours. After the reaction, extract with the same volume of ethyl acetate to remove sugar and enzyme protein impurities. The extract is developed on a G silica gel plate (100mm×25mm), chloroform:methanol:water=7.5:2.5:0.25 as the mobile phase, Observe under 245nm ultraviolet light and carry out qualitative analysis with TLC, the result is as follows figure 2 shown (R f淫羊藿苷标准品 =0.42, 2.R f反应产物 = 0.8) Reaction product R f The value of 0.8 is much larger than the standard R f A value of 0.42 indicates that icariin, which is less polar than icariin, is produced by the enzymatic reaction, and the reaction mechanism is the sam...

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