CN1052239C - Method for prepn. of NTPS - Google Patents
Method for prepn. of NTPS Download PDFInfo
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- CN1052239C CN1052239C CN 94116345 CN94116345A CN1052239C CN 1052239 C CN1052239 C CN 1052239C CN 94116345 CN94116345 CN 94116345 CN 94116345 A CN94116345 A CN 94116345A CN 1052239 C CN1052239 C CN 1052239C
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Abstract
The present invention relates to a method for preparing nerve growth stimulating peptide (NTP<S>), which is a method for extracting the nerve growth stimulating peptide with high bioactivity (NTP<S>) from the brains of healthy young mammals by hollow fibre ultrafiltration. The nerve growth stimulating peptide whose the molecular weight is smaller than 10000 daltons and/or 50000 daltons is extracted by the method. Compared with a conventional dialysis method, the concentration and the activity of the extracted product of the present invention are higher, and the extract can be clinically applied to the treatment of diseases of brain trauma or brain sequela, etc.
Description
That the present invention relates to is NTPS (NTP
s), especially relate to and have the active NTP of stabilate
sTechnical field.
(NTP
s) to prepare traditional technology be to adopt the dialysis method preparation to NTPS, exist yield poorly, active low problem.The cerebrolysin or the like product similar to cerebrolysin that generally are used for the treatment of various nervous system disorderss all are to originate from the pig brain of growing up, thereby prepare NTP in the mammal childhood except that the people
sActive material has not yet to see report.
The object of the present invention is to provide a kind of Hollow Fiber Ultrafiltration technology that adopts to prepare concentration height, output height, active high (NTP
s), and be choose except that the people healthy childhood mammal brain, can make oral liquid, also make injection drug.
Purpose of the present invention can reach by following technical measures: choose except that the people healthy childhood mammal brain, smash → next carry out-20 ℃ of dark freezing → melt back to pieces with any mechanical system and add the dense HCl of 1ml by every 10L liquid, stir evenly → heat 80 ℃, 15min → recovery room temperature, frozen → after melting, centrifugal 3000rpm, 30 minutes → get supernatant liquor → mistake malleation to hold back post
Further specify the collection of illustrative plates of the method for the invention and products thereof below in conjunction with accompanying drawing:
Accompanying drawing 1 is that the uv scan of NTPs of the present invention is analyzed collection of illustrative plates;
Accompanying drawing 2 is the collection of illustrative plates of the HPLC of NTPs of the present invention;
Purpose of the present invention can also reach by following technical measures: the dosage that freezing-thawing method can repeat 1 to for several times, add HCl can be from 1ml to tens of milliliters, purpose is to destroy the colloid equilibrium of homogenate, heating mode be between 60-80 ℃ arbitrary temperature, hold back post system and adopt the Hollow Fiber Ultrafiltration post, molecular cut off≤50000Da reaches≤10000Da, malleation 0.5 * 104Pa, its combination can be single post or the multicolumn series connection is used. NTPS (NTPs) the activity keeping freeze-drying method of preparation :-40 ℃ of quick-frozens, kept 2 hours 30 minutes, be warming up to 0 ℃ half an hour, continue in 1 hour, to be warming up to 37 ℃-38 ℃ after 3 hours and kept the stimulator polypeptide (NTP of preparation 30 hourss) in, molecular weight is polypeptide or the egg blood mixture of 5000-10000Da, has the neuronal survival of promotion and enation, promotes the effect of mitochondria dehydrogenase activity in the neuron.
Can be used for making refreshing brain oral liquid during the NTPS that the present invention prepares (NTPs) molecular cut off≤50000Da, make injecting drug use during molecular cut off≤10000Da. In the time will becoming refreshing brain oral liquid, then to be used for injecting drug use processed with flavor enhancement packing finished product then in preparation process to degerming one step, then go on foot then freeze-drying, packed products in preparation process to packing one.
Purpose of the present invention adopts the hollow fibre filtering technology to prepare stimulator polypeptide, compare with the preparation of traditional handicraft dialysis have the concentration height, output is high, active high characteristics, its comparative result such as following table. (assay adopts improvement Lowry ' s method, the square method patent of determination of activity)
The NTPs that tubular fibre method and dialysis method are produced relatively
| The output of NTPs (ml/g brain) content of peptides (mg/ml) | The tubular fibre method | Dialysis method |
| 2.0 | 1.5 | |
| 1.75 | 0.12 | |
| Activity unit (in/ml) | 87 | 6 |
Healthy childhood of the mammal except that the people of the NTPS (NTPs) of the present invention preparation be selected from as: pig, ox, horse, sheep, dog, exempt from.
The NTPS (NTPs) of the present invention's preparation can be made into oral liquid and intramuscular injection, or splashes into administration with other drug compatibility vein.
Specific implementation method of the present invention is as follows:
NTPS (NTPs) is a kind of medicine that is used for the treatment of nervous system disorders, its preparation method is to adopt the tubular fibre method, choose except that the people healthy childhood mammal brain smash to pieces through any mechanical system, freeze deeply, melt and add HCL, heating, frozen, centrifugal, supernatant liquor, cross malleation and hold back post, degerming, packing, freeze-drying, flow package makes after injecting drug use or the degerming with seasonings, the packing flow process is made oral liquid, they are actually a kind of polypeptide mixture, biological activity is stable, the effect that has the stimulating neuronal enation and support to survive.The NTPS that compares preparation with dialysis method has characteristics such as output height, concentration height, active height.
What the present invention relates to is a kind of medicine for the treatment of nervous system disorders---" Cunaokang " medicine for promoting brain health, it is the NTPS (NTPs) that extracts from brains such as the healthy young mammals except that the people such as sucking pig, new born bovines, molecular weight for≤10000Da and≤50000Da it actual be a kind of polypeptide mixture, biological activity is stable, has the growth of stimulator polypeptide neurite and supports the survival effect.Clinical application can be adopted oral and administered intramuscular, also can splash into other medicines compatibility vein, is demonstrating good effect aspect the treatment nervous system disorders.
Claims (6)
1. the preparation method of a stable NTPS, it is characterized in that described preparation method is, choose except that the people healthy childhood mammal brain, smash to pieces with any mechanical system,-20 ℃ of dark freezing, melting the back adds 1ml in every 10L liquid and adds dense HCl to several milliliters ratio, stir evenly, between 60-80 ℃ of temperature, heat, 15min recovers room temperature, behind frozen the melting, and centrifugal 3000rpm, got supernatant liquor in 30 minutes and cross the Hollow Fiber Ultrafiltration malleation and hold back post, molecular weight cut-off≤50000Da and≤10000Da.
2. the preparation method of NTPS according to claim 1 is characterized in that described preparation method further comprises degerming, packing, freeze-drying, packing.
3. the preparation method of NTPS according to claim 1 is characterized in that described preparation method further comprises with seasonings packing.
4. the preparation method of NTPS according to claim 1, it is characterized in that: freezing-thawing method can repeat 1 to for several times, the dosage that adds HCl can be from 1ml to tens of milliliter, heating mode be between 60-80 ℃ arbitrary temperature, hold back post system and adopt the Hollow Fiber Ultrafiltration post, hold back malleation 0.5 * 10
4Pa, its array mode can be single post or the multicolumn series connection is used.
5. the preparation method of NTPS according to claim 1, it is characterized in that, the active freeze-drying method that keeps of the stimulator polypeptide of preparation :-40 ℃ of quick-frozens, after keeping 2 hours 30 minutes, be warming up to 0 ℃ half an hour, continue in 1 hour, to be warming up to 37 ℃-38 ℃ after 3 hours and kept 30 hours.
6. NTPS preparation method according to claim 1 is characterized in that the molecular weight of the stimulator polypeptide for preparing is 5000-10000Da.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 94116345 CN1052239C (en) | 1994-10-10 | 1994-10-10 | Method for prepn. of NTPS |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 94116345 CN1052239C (en) | 1994-10-10 | 1994-10-10 | Method for prepn. of NTPS |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1120547A CN1120547A (en) | 1996-04-17 |
| CN1052239C true CN1052239C (en) | 2000-05-10 |
Family
ID=5037884
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 94116345 Expired - Lifetime CN1052239C (en) | 1994-10-10 | 1994-10-10 | Method for prepn. of NTPS |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1052239C (en) |
-
1994
- 1994-10-10 CN CN 94116345 patent/CN1052239C/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| CN1120547A (en) | 1996-04-17 |
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Expiration termination date: 20141010 Granted publication date: 20000510 |