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Hepatocyte growth-promoting factor and preparation and application thereof

A hepatocyte auxin and preparation technology, applied in the field of new hepatocyte auxin, can solve the problems of long heating time, cumbersome extraction process, and loss of active substance content, so as to save energy, prevent membrane damage, and avoid degradation The effect of pressing the substance

Active Publication Date: 2014-01-29
GUANGZHOU YIPINHONG PHARMA +4
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantages of this preparation method are: the stirring time is too long, and the heating time is too long under the high temperature condition of 95° C., which will easily cause the decomposition and inactivation of the active polypeptide in the hepatocyte growth-stimulating hormone; The salt eluent is eluted and then desalted. These operations will lose the content of active substances, resulting in low activity of the extract and low extraction rate.
This extraction method adopts the process of repeated freezing and thawing, which is very easy to produce antihypertensive substances, and the temperature of thermal denaturation in this process is too high, which will also cause the inactivation of polypeptide substances

Method used

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  • Hepatocyte growth-promoting factor and preparation and application thereof
  • Hepatocyte growth-promoting factor and preparation and application thereof
  • Hepatocyte growth-promoting factor and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1 Extraction of hepatocyte growth-stimulating factor

[0063] (1) Material selection: the raw material is fresh suckling pig liver slaughtered on the same day; the weight of a single whole pig liver shall not exceed 140 grams;

[0064] (2) Tissue processing: After the fascia and pedicles of suckling pig liver are removed, wash with pure water; cut into 2-3 pieces with a stainless steel knife to facilitate homogenization;

[0065] (3) Mix with water for injection at room temperature, the speed of homogenate is 800~1000 rpm, the time is: 5min, the ratio is water: pig liver mass ratio is 11:1; Freeze to 0°C, then freeze to -35°C at a rate of 4°C / min;

[0066] (4) After thawing the homogenate that has been frozen at -10~-25℃ for at least 7 days, heat it to 83℃~85℃, remove the heating source immediately, put it into a filter cloth and filter it naturally, and adjust the pH value of the filtrate with NaOH solution to 6.5, the speed of thawing and heating is 1~2℃ / min...

Embodiment 2

[0075] (1) Material selection: the raw material is fresh suckling pig liver slaughtered on the same day; the weight of a single whole pig liver shall not exceed 140 grams;

[0076] (2) Tissue processing: After the fascia and pedicles of suckling pig liver are removed, wash with pure water; cut into 2-3 pieces with a stainless steel knife to facilitate homogenization;

[0077] (3) mix with water for injection at room temperature, the speed of homogenate is 800~1000 rpm, the time is 5 minutes, the ratio is water: pig liver mass ratio is 115: 1; Freeze at room temperature to -5°C, and then freeze to -35°C at a rate of 5°C / min.

[0078] (4) After thawing the homogenate that has been frozen at -35°C for at least 7 days, heat it to 83°C-85°C and immediately remove the heating source, put it into a filter cloth and filter naturally, and adjust the pH value of the filtrate to 6.5 with NaOH solution; thaw And the heating speed is raised to 0°C at a rate of 2°C / min, and then heated to ...

Embodiment 3

[0087] (1) Material selection: the raw material is fresh suckling pig liver slaughtered on the same day; the weight of a single whole pig liver shall not exceed 140 grams;

[0088] (2) Tissue processing: After the fascia and pedicles of suckling pig liver are removed, wash with pure water; cut into 2-3 pieces with a stainless steel knife to facilitate homogenization;

[0089] (3) Mix with water for injection at room temperature, the speed of homogenate is 800-1000 rpm, the time is 5 minutes, the mass ratio of water and pig liver is 11:1; the homogenate is frozen from room temperature at a speed of 18°C / min to -2°C, and then frozen to -28°C at a rate of 8°C / min;

[0090] (4) After thawing the homogenate that had been frozen at -30°C for at least 7 days, heat it to 83°C and immediately remove the heating source, put it into a filter cloth and filter it naturally, and adjust the pH value of the filtrate to 6.5 with NaOH solution;

[0091] (5) The solution adjusted to pH 6.5 is f...

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Abstract

The invention relates to a hepatocyte growth-promoting factor. A preparation method for the hepatocyte growth-promoting factor comprises the following steps of: selecting materials; treating tissues; mixing the materials and injection water at room temperature, uniformly stirring, and freezing homogenate liquid to the temperature of between -20 and -35 DEG C; unfreezing the homogenate liquid which is subjected to freezing storage for at least seven days, then heating to the temperature of between 83 and 85 DEG C, immediately removing a heating source, filtering, and adjusting the pH value to be 6.5; filtering by using a hollow fiber column with the molecular weight cutoff of 20,000, then filtering by using an ultrafiltration membrane with the molecular weight cutoff of 10,000, freezing ultrafiltrate to the temperature of between -20 and -30 DEG C, and storing in a sealing way; and unfreezing the ultrafiltrate which is subjected to freezing storage for at least seven days, filtering, concentrating, filtering the concentrated solution by using a hollow fiber column with the molecular weight cutoff of 10,000, degerming by using a filtration membrane with the aperture of 0.22 micrometer, performing split charging, and storing at the temperature of below -18 DEG C. The invention also relates to a preparation and application of the hepatocyte growth-promoting factor. The hepatocyte growth-promoting factor has high activity and high yield and is safely used.

Description

technical field [0001] The invention relates to a hepatocyte growth-stimulating hormone, in particular to a new hepatocyte growth-stimulating hormone, its preparation and application. Background technique [0002] Hepatocyte growth-stimulating hormone is a small molecular weight polypeptide mixture extracted from fresh suckling pig liver, with a molecular weight below 10,000. Its pharmacological effects are mainly to stimulate the DNA synthesis of normal liver cells, promote the regeneration of liver cells, protect the damaged liver cells, promote the recovery of liver function, and also regulate the body's immune function and anti-hepatic fibrosis. It is clinically used to treat severe hepatitis, chronic active hepatitis and liver cirrhosis. [0003] At present, there have been many patents and literature reports on hepatocyte growth-stimulating hormone, a biological agent drug. [0004] In the patent ZL03116052.2, a hepatocyte growth-promoting hormone injection and its p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/00A61P1/16A61K35/407
Inventor 孔祥平陈光明张宜俊杨富强张海松陈阳述
Owner GUANGZHOU YIPINHONG PHARMA
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