CN1048629C

CN1048629C - Brain hormone production method

Info

Publication number: CN1048629C
Application number: CN93103863A
Authority: CN
Inventors: 荣永海
Original assignee: NEW BIOCHEMICAL TECHNOLOGY INST WEST DISTRICT BEIJING CITY
Current assignee: NEW BIOCHEMICAL TECHNOLOGY INST WEST DISTRICT BEIJING CITY
Priority date: 1993-04-10
Filing date: 1993-04-10
Publication date: 2000-01-26
Anticipated expiration: 2013-04-10
Also published as: CN1077376A

Abstract

The present invention relates to a method for extracting brain hormone from animal brains, which is formed by that water is added in a pig brain to be uniformly mixed, a pH value is adjusted by Ca (OH) 2, the pancreatin of a pig or a cattle, toluene and distilled water are added to be stirred at the temperature of 37 DEG C to 50DEG. C, the pH value is adjusted by phosphoric acid, the pH value is adjusted via a series of boiling, activated carbon is decolored, and the water is dialyzed, filtered, separately filled and sterilized by a dialysis bag. The content of klinitamin contained in the present invention reaches 35 mg/ml to 38 mg/ml via analysis, and the extracted brain hormone has the same effect as the brain hormone generated by Austria via clinical tests.

Description

From Medulla sus domestica, extract the method for brain protein hydrolyzate

The present invention relates to a kind of method of from animal brain, extracting the brain protein hydrolyzate, particularly a kind of method of from Medulla sus domestica, extracting the brain protein hydrolyzate.

Cerebrolysin is in recent years by the big pharmaceutical factory of the Austrian Yi Biwei a kind of new drug that comes across the market newly developed, its registered trade mark is Cerebrolysin, it is by the aqueous solution of protein-free standardization organ specific amino acid mixture, contains free amino acid and molecular weight at the low molecular peptide below 10,000.As the aminoacid (isoleucine, leucine, lysine, methionine etc.) and the nonessential aminoacid of human body (alanine, arginine etc.) that contain needed by human also have glutamine, asparagine etc.For the organic brain psychosyndrome of treatment, the compensatory deficiency of cerebrovascular, the neurasthenia state, child's cerebral hypoplasia, epilepsy, apoplexy, encephalitis, cerebral concussion etc. have favorable effects.

But the extracting method about cerebrolysin is in the secret stage always, and openly publication yet there are no its extracting method of report both at home and abroad.

The object of the present invention is to provide a kind of method of from Medulla sus domestica, extracting the brain protein hydrolyzate, by clinical proof, the extract of gained, its effect is identical with cerebrolysin.

Embodiment of the present invention are as follows: with the homogenate of fresh Medulla sus domestica adding distil water, it is 7-9 that the hydro-oxidation calcium solution is transferred pH value, add hydrolytic reagent such as pig or bovine trypsin, adding preservative agent such as toluene, distilled water, stirred 10-20 hour down at 37-50 ℃, transferring pH value with phosphoric acid is 5.7-5.8, boils, filter, it is 6-7 that filtrate is transferred pH value with hydrochloric acid, concentrating under reduced pressure, sub-cooled, filter, filtrate concentrates, and it is 7-9 that the reuse sodium hydroxide is transferred pH value, be evaporated to dried, add water again, filter, it is 4-5 that filtrate is transferred PH with hydrochloric acid, use activated carbon decolorizing, after the filtering and concentrating with interception be 10,000 bag filter to water dialysis 24-28 hour, in extracellular fluid dialysis, add sodium sulfite, filter the packing sterilization and form.

Following preference elaborates to the present invention, but does not limit the scope of the invention.

Example 1,1 kilogram fresh Medulla sus domestica is placed in general 10 liters of containers, add 1 liter of distilled water and 5N aqua calcis, transferring pH value is 8.0, adds 8 gram Pancreas Sus domestica enzyme and several toluene and 4 liters of distilled water, in 45 ℃ water-bath, stirred 10 hours, transferring pH value with phosphoric acid solution is 5.7, boils half an hour, filter, filtering residue discards, and filtrate is transferred pH value to 6.5 with 5N hydrochloric acid, is concentrated to 1500 milliliters under 70 ℃ of 500m/mHg column temperature, under 8 ℃, left standstill 24 hours, filter, filtering residue discards, and filtrate is concentrated to 250 milliliters under 70 ℃ on 500m/mHg post, transfer PH to 8 with the 5N sodium hydroxide, be concentrated into driedly under 70 ℃ on 500m/mHg post, 2000 milliliters of dissolvings of adding distil water are again filtered, filtering residue discards, and filtrate is transferred PH to 4.5 with 5HN hydrochloric acid.Adding 100 gram activated carbon decolorizings heat half an hour down at 80 ℃ again, filter, and filtering residue discards, and filtrate is concentrated to 1000 milliliters on 70 ℃ on 500m/mHg post, are that 10,000 bag filter was dialysed 24 hours to water with interception, then, add 0.06%NaHSO in extracellular fluid dialysis ₃Refilter, filtering residue discards, filtrate packing 2.5 or 10 milliliters peace bottle, 100 ℃ of sterilizations, altogether 1000 milliliters, the mixed amino acid content is 40mg/ml by analysis.

Example 2, except that replacing the Pancreas Sus domestica enzyme with bovine trypsin, all the other conditions are identical with example 1, make 1000 milliliters of finished products, and the content of kilnitamin is 38%.

The advantage of the inventive method:

(1) method is simple and reliable.

(2) adopt common apparatus and the most frequently used chemical reagent.

(3) extraction effect is good, and raw material sources are abundant.

Claims

1, a kind of method of extracting the brain protein hydrolyzate from Medulla sus domestica is characterized in that:

(1) with the Medulla sus domestica adding distil water, with Ca (OH) ₂Accent PH is 7-9,

(2) add pig or bovine trypsin, its amount is 8 gram/1kg Medulla sus domestica, reaches toluene, distilled water 37-50 ℃ of following the stirring 10-20 hour,

(3) transferring PH with phosphoric acid is 5.7-5.8,

(4) boil, filter, it is 6-7 that filtrate is transferred pH value with hydrochloric acid, concentrating under reduced pressure, sub-cooled, filtration, reconcentration, transferring pH value with NaOH is 7-9, be evaporated to dried,

(5) add water filtration, it is 4-5 that filtrate is transferred PH with hydrochloric acid, and with activated carbon decolorizing, filtration, concentrating the back is that 10,000 bag filter carries out water was dialysed 24 hours with interception, adds NaHSO in extracellular fluid dialysis ₃, filter the packing sterilization and form.