A kind of preparation method of polymyocyte

Abstract

The invention belongs to a preparation method of poly i-c and belongs to the technical field of biomedicine. The preparation method includes: utilizing inosine diphosphate to prepare polyinosinic acid, and using cytidine diphosphate to prepare polycytidylic acid; utilizing the prepared polyinosinic acid to react with the polycytidylic acid to obtain a high-purity poly i-c product. Poly i-c is prepared through self-developed high-purity polyinosinic acid and polycytidylic acid, so that prepared poly i-c is high in purity, and treatment difficulty in the process of preparing is lowered.

Description

technical field

[0001] The invention relates to the technical field of biomedicine, in particular to a preparation method of polymyocytes. Background technique

[0002] Polyinosinic acid cytidylic acid (referred to as polyinosinic acid, PIC) is a safe and efficient innate immune activator, with broad-spectrum anti-virus, inhibiting tumor cell growth, enhancing the body's immunity and other functions, the global market capacity potential Estimated at tens of billions of dollars per year. In clinical medicine, it can be used for the treatment of viral diseases such as chronic hepatitis B, epidemic hemorrhagic fever, epidemic encephalitis Japanese, and viral keratitis. The adjuvant treatment of breast cancer, prostate cancer and other tumors has been in Phase III clinical trials experimental stage. In veterinary clinics, it can be used for the prevention and treatment of viral diseases such as foot-and-mouth disease, avian influenza, and swine fever, and can also improve the ...

Examples

Embodiment 1

[0031] A preparation method of polymyocytes, comprising the following steps:

[0032] 1. Preparation of cytidine diphosphate CDP solution

[0033] 1) Phosphorylation, put 300 parts by weight of yeast into the phosphorylation reaction tank and stir, heat in a water bath to 31 ° C, stop heating, put in 5 parts by weight of magnesium chloride, 30 parts by weight of potassium dihydrogen phosphate, and 20 parts by weight of white sugar, and control the temperature at 31 °C. react at ℃ for 20 minutes, then add 30 parts by weight of cytidylic acid, and react for 3 hours. When the content of cytidine triphosphate is greater than 85%, the reaction is completed. Immediately adjust the pH to 2 to terminate the reaction, and at the same time, cool with refrigerated water;

[0034] 2) Chromatographic purification, the phosphorylated product after cooling in step 1) is subjected to plate and frame filtration, chromatography, adsorption, washing, elution and decolorization sodium filtration;...

Embodiment 2

[0057] A preparation method of polymyocytes, comprising the following steps:

[0058] 1. Preparation of cytidine diphosphate CDP solution

[0059] 1) Phosphorylation, put 200 parts by weight of yeast into the phosphorylation reaction tank and stir, heat in a water bath to 30°C, stop heating, put in 2 parts by weight of magnesium chloride, 20 parts by weight of potassium dihydrogen phosphate, and 20 parts by weight of white sugar, and control the temperature at 31 React at ℃ for 10 minutes, then add 30 parts by weight of cytidylic acid, and react for 2 hours. When the content of cytidine triphosphate is greater than 85%, the reaction is completed. Immediately adjust the pH to 2 to terminate the reaction, and at the same time, cool with refrigerated water;

[0060] 2) Chromatographic purification, the phosphorylated product after cooling in step 1) is subjected to plate and frame filtration, chromatography, adsorption, washing, elution and decolorization sodium filtration;

[0...

Embodiment 3

[0083] A preparation method of polymyocytes, comprising the following steps:

[0084] 1. Preparation of cytidine diphosphate CDP solution

[0085] 1) Phosphorylation, put 500 parts by weight of yeast into the phosphorylation reaction tank and stir, heat in a water bath to 31 ° C, stop heating, put in 10 parts by weight of magnesium chloride, 50 parts by weight of potassium dihydrogen phosphate, and 50 parts by weight of white sugar, and control the temperature at 33 React at ℃ for 20 minutes, then add 30 parts by weight of cytidylic acid, and react for 5 hours. When the content of cytidine triphosphate is greater than 85%, the reaction is completed. Immediately adjust the pH to 2 to terminate the reaction, and at the same time, cool with refrigerated water;

[0086] 2) Chromatographic purification, the phosphorylated product after cooling in step 1) is subjected to plate and frame filtration, chromatography, adsorption, washing, elution and decolorization sodium filtration;

...

technical field

[0001] The invention relates to the technical field of biomedicine, in particular to a preparation method of polymyocytes. Background technique

[0002] Polyinosinic acid cytidylic acid (referred to as polyinosinic acid, PIC) is a safe and efficient innate immune activator, with broad-spectrum anti-virus, inhibiting tumor cell growth, enhancing the body's immunity and other functions, the global market capacity potential Estimated at tens of billions of dollars per year. In clinical medicine, it can be used for the treatment of viral diseases such as chronic hepatitis B, epidemic hemorrhagic fever, epidemic encephalitis Japanese, and viral keratitis. The adjuvant treatment of breast cancer, prostate cancer and other tumors has been in Phase III clinical trials experimental stage. In veterinary clinics, it can be used for the prevention and treatment of viral diseases such as foot-and-mouth disease, avian influenza, and swine fever, and can also improve the ...

Examples

Embodiment 1

[0031] A preparation method of polymyocytes, comprising the following steps:

[0032] 1. Preparation of cytidine diphosphate CDP solution

[0033] 1) Phosphorylation, put 300 parts by weight of yeast into the phosphorylation reaction tank and stir, heat in a water bath to 31 ° C, stop heating, put in 5 parts by weight of magnesium chloride, 30 parts by weight of potassium dihydrogen phosphate, and 20 parts by weight of white sugar, and control the temperature at 31 °C. react at ℃ for 20 minutes, then add 30 parts by weight of cytidylic acid, and react for 3 hours. When the content of cytidine triphosphate is greater than 85%, the reaction is completed. Immediately adjust the pH to 2 to terminate the reaction, and at the same time, cool with refrigerated water;

[0034] 2) Chromatographic purification, the phosphorylated product after cooling in step 1) is subjected to plate and frame filtration, chromatography, adsorption, washing, elution and decolorization sodium filtration;...

Embodiment 2

[0057] A preparation method of polymyocytes, comprising the following steps:

[0058] 1. Preparation of cytidine diphosphate CDP solution

[0059] 1) Phosphorylation, put 200 parts by weight of yeast into the phosphorylation reaction tank and stir, heat in a water bath to 30°C, stop heating, put in 2 parts by weight of magnesium chloride, 20 parts by weight of potassium dihydrogen phosphate, and 20 parts by weight of white sugar, and control the temperature at 31 React at ℃ for 10 minutes, then add 30 parts by weight of cytidylic acid, and react for 2 hours. When the content of cytidine triphosphate is greater than 85%, the reaction is completed. Immediately adjust the pH to 2 to terminate the reaction, and at the same time, cool with refrigerated water;

[0060] 2) Chromatographic purification, the phosphorylated product after cooling in step 1) is subjected to plate and frame filtration, chromatography, adsorption, washing, elution and decolorization sodium filtration;

[0...

Embodiment 3

[0083] A preparation method of polymyocytes, comprising the following steps:

[0084] 1. Preparation of cytidine diphosphate CDP solution

[0085] 1) Phosphorylation, put 500 parts by weight of yeast into the phosphorylation reaction tank and stir, heat in a water bath to 31 ° C, stop heating, put in 10 parts by weight of magnesium chloride, 50 parts by weight of potassium dihydrogen phosphate, and 50 parts by weight of white sugar, and control the temperature at 33 React at ℃ for 20 minutes, then add 30 parts by weight of cytidylic acid, and react for 5 hours. When the content of cytidine triphosphate is greater than 85%, the reaction is completed. Immediately adjust the pH to 2 to terminate the reaction, and at the same time, cool with refrigerated water;

[0086] 2) Chromatographic purification, the phosphorylated product after cooling in step 1) is subjected to plate and frame filtration, chromatography, adsorption, washing, elution and decolorization sodium filtration;

...