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Cerebrolysin hydrolysate injection and preparation method thereof

A technology for cerebroprotein hydrolysate and injection, which is applied in the field of cerebroprotein hydrolysate injection and its preparation, and achieves the effects of high guaranteed value, improved medication safety, and stable medicinal effect.

Active Publication Date: 2014-09-17
GUANGZHOU YIPINHONG PHARMA +4
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the defect that additional corresponding amino acids need to be added in the existing cerebroprotein hydrolyzate preparation technology, and provide a cerebroprotein hydrolyzate injection and a preparation method thereof. The cerebroprotein hydrolyzate injection prepared by the preparation method has small Molecular peptide reversed-phase peptide map chromatographic identification and the similarity of the reference substance ≥ 0.90, no need to add exogenous amino acids, completely rely on pig brain hydrolysis

Method used

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  • Cerebrolysin hydrolysate injection and preparation method thereof
  • Cerebrolysin hydrolysate injection and preparation method thereof
  • Cerebrolysin hydrolysate injection and preparation method thereof

Examples

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Effect test

Embodiment 1

[0054] Take 100kg of brain tissue (quarantine qualified) after removing blood vessels, fat and other impurities, add 1 times the volume of purified water to homogenate, keep warm at 82°C for 15 minutes, cool; adjust pH to 1.8 with hydrochloric acid, hydrolyze with pepsin, keep warm at 42°C for 3 hours, Adjust the pH to 7.6 with sodium hydroxide, hydrolyze with trypsin, incubate at 42°C for 2 hours, and collect the supernatant by centrifugation; adjust the pH to 2 with hydrochloric acid, incubate at 105°C for 30 minutes, then adjust the pH to 8.8 with sodium hydroxide; cool down, extract the supernatant, 10kd membrane ultrafiltration to remove macromolecules, collect 100L of filtrate; apply column chromatography, wash column with purified water, analyze, collect eluate, adjust pH to 8.0-9.0, concentrate to 50L with DUS-8040C ion membrane, 0.2μm membrane Filtrate to obtain the stock solution, freeze and store below -15°C; thaw the stock solution, blend, add 0.01wt% sodium bisulfi...

Embodiment 2

[0069] Brain tissue (qualified for inspection and quarantine) removes impurities such as blood vessels and takes 100kg, add 2 times the volume of purified water to homogenate, keep warm at 85°C for 20 minutes, cool, adjust pH to 2.0 with hydrochloric acid, hydrolyze with pepsin, keep warm at 42°C for 4 hours, Adjust pH to 7.8 with sodium hydroxide, hydrolyze with trypsin, incubate at 42°C for 2 hours, collect supernatant by centrifugation; adjust pH to 2 with hydrochloric acid, incubate at 110°C for 30 minutes, adjust pH to 9.0 with sodium hydroxide; cool, extract supernatant, 10kd membrane Remove macromolecules by ultrafiltration, collect 150L of filtrate; apply column chromatography, wash column with purified water, analyze, collect eluate, adjust pH to 8.0-9.0, concentrate 50L with DUS-040C ion membrane, filter with 0.2μm membrane to obtain Stock solution, frozen and stored below -15°C; stock solution thawed, prepared, added 0.02wt% sodium bisulfite, adjusted to pH 6.9-7.5, ...

Embodiment 3

[0076] Brain tissue (qualified for inspection and quarantine) removes impurities such as blood vessels and takes 100kg, add 2 times the volume of purified water to homogenate, keep warm at 85°C for 30 minutes, cool, adjust pH to 3.0 with hydrochloric acid, hydrolyze with pepsin, keep warm at 35°C for 5 hours, Adjust the pH to 7.2 with sodium hydroxide, hydrolyze with trypsin, incubate at 45°C for 3 hours, and centrifuge to collect the supernatant; 10kd membrane ultrafiltration to remove macromolecules, collect 150L of filtrate; apply column chromatography, wash the column with purified water, analyze, collect eluate, adjust pH to 8.0-9.0, concentrate 50L with DUS-040C ion membrane, and filter with 0.2μm membrane , to obtain the original solution, and freeze it below -15°C; thaw the original solution, prepare it, add 0.02wt% sodium metabisulfite, adjust the pH to 6.9-7.5, 8-10kd membrane ultrafiltration, 0.2μm membrane filtration, filling; 115°C, 30 minutes , leak detection, et...

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Abstract

The invention provides a cerebrolysin hydrolysate injection and a preparation method thereof. The preparation method comprises the following steps: taking pig brain, removing impurities, and adding purified water for homogenizing; hydrolyzing homogenate through pepsin and pancreatin, and collecting a clear liquid; regulating the pH of the clear liquid to be 1.7-3, heating to 100-110 DEG C, insulating for 15-45min, then regulating pH to 8.0-9.0; conducting ultrafiltration through a 8-10kd membrane, and collecting filtrate; conducting column chromatography to the filtrate, collecting an eluent, regulating ph to 8.0-9.0, concentrating through an anion-exchange membrane, filtering through a 0.2mu m membrane to obtain a stock liquid; and freezing the stock liquid, then defreezing, adding an antioxidant, regulating the pH to be 6.9-7.5, and filtering respectively through a 8-10kd membrane and a 0.2mu m membrane. The polypeptide in the obtained cerebrolysin hydrolysate injection is similar to that of a control drug being more than or equal to 0.90, and the obtained cerebrolysin hydrolysate injection contains 16 amino acids, the nitrogen amount of total amino acids accounts for more than or equal to 85% of total nitrogen content, additional amino acids are not required, and the amino acids can be prepared completely by means of pig brain, thus being stable and safe.

Description

technical field [0001] The invention relates to the field of biological preparations, in particular to a cerebroprotein hydrolyzate injection and a preparation method thereof. Background technique [0002] Cerebroprotein hydrolyzate injection products are prepared from raw materials that have been inspected, quarantined, extracted, refined, sterilized and other production processes; the main components of cerebroprotein hydrolyzate are low-molecular-weight peptides and free amino acids. It is easy to enter the brain nerve cells through the blood-brain barrier. Low-molecular-weight peptides have the same effect as naturally occurring neurotrophic factors, which can induce neuron differentiation, maintain normal nerve cell functions, increase neurotransmitter production and synaptic transmission, and protect Nerve cells can be protected from various damages, regulate human brain metabolism, neurotransmission and nerve growth; can increase the activity of glucolytic enzymes, in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/01A61K35/30A61K9/08A61K47/34A61P3/02A61P25/00A61P9/10
Inventor 张莉李捍雄
Owner GUANGZHOU YIPINHONG PHARMA
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