CN1736475A - Troxerutin brain protein hydrolysate for injection and preparation process thereof - Google Patents
Troxerutin brain protein hydrolysate for injection and preparation process thereof Download PDFInfo
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- CN1736475A CN1736475A CN 200510094044 CN200510094044A CN1736475A CN 1736475 A CN1736475 A CN 1736475A CN 200510094044 CN200510094044 CN 200510094044 CN 200510094044 A CN200510094044 A CN 200510094044A CN 1736475 A CN1736475 A CN 1736475A
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Abstract
The invention relates to a troxerutin brain protein hydrolysate for injection and preparation process, for treating cerebrovascular diseases and neurous system diseases, which is prepared from Troxerutin and Cerebrolysin by the weight ratio of 50-100 : 1-20, preferably 80:1, the pH of the composition is between 5.5-7.5. Its preparing process is also disclosed in the invention.
Description
Technical field
The present invention relates to a kind of injection of troxerutin and encephalic protein hydrolysate, is the compound preparation that is used for the treatment of cerebrovascular and central nervous system disease.
Background technology
It is multiple that cerebrovascular disease mainly contains cerebral thrombosis, cerebral embolism, chamber barrier property infraction, cerebral hemorrhage, cerebral arteriosclerosis etc., the symptom of these diseases performance has symptoms such as complete or incomplete aphasia, quadriplegia, the pain sensation or thought disturbance, insomnia, stupefied, hypomnesis, tinnitus, brain function decline, these symptoms have been brought great misery to patient, existing medicine cure rate is low, can not effect a radical cure, can only relief of symptoms, curative effect is slow.
The related preparations of troxerutin has been widely used in clinical, has the erythrocyte of inhibition and platelet aggregation, prevents thrombosis, increases oxygen saturation in the blood, effects such as microcirculation improvement.Simultaneously the troxerutin vascular injury that can cause medmain, Kallidin I increases capillary resistance, reduces capillary permeability, can prevent the edema that vascular permeability raises and causes.Be applicable to hemiplegia, aphasia and preinfarction syndrome, arteriosclerosis, central serous chorioretinopathy, thrombophlebitis, varicosis, vascular permeability due to cerebral thrombosis and the cerebral embolism edema that causes etc. that raises.Single invalid to the protection that improves brain cell physiological function and cranial nerve with this product.
Cerebrolysin Vial (Cerebroprotein Hydrolysate) is the nonprotein micromolecule polypeptide through separation and purification, includes the free amino acid of 16 kinds of needed by human, has kept constant ratio between its each seed amino acid.Have good cerebral protection and cranial nerve Nutrition, clinical practice proves that Cerebrolysin Vial can safeguard effectively that the central nervous system avoids the infringement of noxious substance, can improve the anti-anoxia ability of brain.The auxiliary treatment that is used for symptoms such as apoplexy acute stage, convalescent period hemiplegia, aphasia, dementia, drowsiness, weak, giddy clinically.Simultaneously, this product reduces plasma fibrinogen, platelet aggregation rate and increasing high density fat egg self-applying in addition.The brain metabolism is strengthened, improve the blood supply state of brain cell, neurotransmitter is played a role better, for the senile dementia curative effect that improves significantly.This product is singly used improving cerebral infarction, apoplexy, and cerebral arteriosclerosis does not have direct effect, but as a kind of nutrient substance for and brain cell, cranial nerve, thereby improve the brain function metabolism.
Summary of the invention
The purpose of this invention is to provide a kind of reasonable recipe, preparation technology is simple, the injection of troxerutin and encephalic protein hydrolysate of drug safety and preparation technology thereof are can remedy the deficiency of each monomer medicine injection of troxerutin and Cerebrolysin Vial on therapeutic effect.
Injection of troxerutin and encephalic protein hydrolysate of the present invention is made up of troxerutin and Cerebrolysin Vial, the two weight proportion is: troxerutin 50～100: Cerebrolysin Vial 1～20, wherein Cerebrolysin Vial content is by free amino acid total nitrogen (down together), the pH value 5.5～7.5 of compositions, character is yellow to unformed solid of sundown or powder, dissolving back yellowly supernatant liquid, described raw material are suitable for the proportioning preparation by the best by following weight and are: 80: 1 (troxerutins: the Cerebrolysin Vial total nitrogen)
The preparation technology of injection of troxerutin and encephalic protein hydrolysate of the present invention is as follows: troxerutin and Cerebrolysin Vial stock solution are mixed in proportion, add an amount of water for injection to prescribed volume, press amount of liquid 0.07% and add the injection stage medicinal charcoal, stirring and adsorbing is 30 minutes under the room temperature, earlier through coarse filtration, measures qualified after the microporous filter membrane fine straining of 0.22 μ m, get filtrate, packing, put in the freeze drying box (pre-freeze to-40C), insulation is 2 hours under this temperature; Start the water vessel switch, start vacuum pump, under 4Pa, gradually with per hour 1 ℃ improve temperature.When arriving room temperature, continued evacuation 3 hours.Lyophilizing finishes, and opens the elevator of freeze dryer, and plug is pressed in the cillin bottle fully, feeds filtrated air then, and the good product Zha Gai of taking-up lyophilizing that unpacks gets final product.
Troxerutin of the present invention and Cerebrolysin Vial be anticoagulant effectively, the formation of anti-tampon, again because of being rich in multiple free amino acid and micromolecule polypeptide, can see through blood brain barrier and act on nervus centralis in many ways, directly provide neurotrophic factor, improve the neuron metabolism, induce and promote the reparation and the regeneration of neurocyte to brain cell, induce neurocyte to form new aixs cylinder simultaneously, connect thereby form new nerve.Two recurrence due to taking drug sides produce the cooperative compensating effect, have ion channel gate effect, produce the effect of class calcium antagonist medicine treatment cerebrovascular disease.Simultaneously, can also strengthen various neurotrophy materials and enter blood brain barrier, improve the metabolic function of cranial nerve tissue.Because this product has stronger antioxidation anti-radical action, can repair cerebrovascular inwall inner film injury rapidly, also help the stable integrity that keeps cerebrovascular integrity and metabolic process, significant for the treatment and the rehabilitation of all kinds of cerebrovascular disease.For clinical medication to different patients, different modes of administration and drug combination are provided convenience.This compound preparation can be treated acute and chronic cerebrovascular disease such as being used for the treatment of cerebral thrombosis, cerebral embolism, brain spasm, and the edema that causes of back something lost such as cerebrovascular disease due to the craniocerebral trauma and caused disordered brain function.
Description of drawings
Fig. 1 is tame rabbit ear edge irritation test pathological section figure.
Fig. 2 is rabbit quadriceps femoris zest pathological section figure.
The specific embodiment
The preparation of embodiment 1 injection of troxerutin and encephalic protein hydrolysate
Take by weighing troxerutin 80g, Cerebrolysin Vial stock solution 200g (total nitrogen is in 5.0mg/ml), water for injection 700g, mix, add the injection water to 1000ml, press amount of liquid 0.07% and add the injection stage medicinal charcoal, stirring and adsorbing is 30 minutes under the room temperature, earlier through coarse filtration, measure qualified after the microporous filter membrane fine straining of 0.22 μ m, divide in the cillin bottle of packing into, put (pre-freeze is to-40 ℃) in the freeze drying box, insulation is 2 hours under this temperature; Start the water vessel switch, start vacuum pump, under 4Pa, gradually with per hour 1 ℃ improve temperature.When arriving room temperature, continued evacuation 3 hours.Lyophilizing finishes, and opens the elevator of freeze dryer, and plug is pressed into fully, feeds filtrated air then, and the good product Zha Gai of taking-up lyophilizing that unpacks gets final product.
The preparation of embodiment 2 injection of troxerutin and encephalic protein hydrolysate
Take by weighing troxerutin 200g, Cerebrolysin Vial stock solution 500g (total nitrogen is in 5.0mg/ml), water for injection 700g, mix, add an amount of water for injection to 2300ml, press amount of liquid 0.07% and add the injection stage medicinal charcoal, stirring and adsorbing is 30 minutes under the room temperature, earlier through coarse filtration, measure qualified after the microporous filter membrane fine straining of 0.22 μ m, packing is put (pre-freeze is to-40 ℃) in the freeze drying box, and insulation is 3 hours under this temperature; Start the water vessel switch, start vacuum pump, under 4Pa, gradually with per hour 1 ℃ improve temperature.When arriving room temperature, continued evacuation 3 hours.Lyophilizing finishes, and opens the elevator of freeze dryer, and plug is pressed in the cillin bottle fully, feeds filtrated air then, and the good product Zha Gai of taking-up lyophilizing that unpacks gets final product.
The preparation of embodiment 3 injection of troxerutin and encephalic protein hydrolysate
Take by weighing troxerutin 400g, Cerebrolysin Vial stock solution 1000g (total nitrogen is in 5.0mg/ml), water for injection 700g, mix, add the injection water to 5000ml, press amount of liquid 0.07% and add the injection stage medicinal charcoal, stirring and adsorbing is 30 minutes under the room temperature, earlier through coarse filtration, measure qualified after the microporous filter membrane fine straining of 0.22 μ m, packing is put (pre-freeze is to-40 ℃) in the freeze drying box, and insulation is 3 hours under this temperature; Start the water vessel switch, start vacuum pump, under 4Pa, gradually with per hour 1 ℃ improve temperature.When arriving room temperature, continued evacuation 3 hours.Lyophilizing finishes, and opens the elevator of freeze dryer, and plug is pressed in the cillin bottle fully, feeds filtrated air then, and the good product Zha Gai of taking-up lyophilizing that unpacks gets final product.
Embodiment 5 safety testings
This compound preparation has carried out the specific safety test, for clinical application safety provides objectively foundation of science.Its test method and result are as follows respectively:
Trial drug: injection of troxerutin and encephalic protein hydrolysate of the present invention, yellow freeze-dried powder, specification: 80mg troxerutin and 1.0mg total nitrogen.Provide by the gloomy medical biotechnology company limited of Hainan Hewlett-Packard.
Experimental animal: 1 of the big white doe of ear of Japan, body weight 2.3kg, one-level is provided by Hainan Province's Experimental Animal Center.The animal quality certification number: moving word 30-003 number of doctor.60 of Cavia porcelluss, body weight 230～300g is provided and is provided by Animal Experimental Study center, Hubei Province.Credit number: SCXK (Hubei Province) 2003-2005.
One, hemolytic test
The preparation of (1) 2% Sanguis Leporis seu oryctolagi cell suspension: get rabbit cardiac blood 15ml, remove fibrin with bead, synthetic defibrinated blood, in the immigration graduated centrifuge tube, add normal saline, shake up, centrifugal, lose supernatant, till directly being washed till supernatant and being water white transparency with normal saline, with normal saline erythrocyte is diluted to 2% suspension then.
(2) test method: troxerutin and encephalic protein hydrolysate of the present invention, get 7 in test tube, press table 1 and add various liquid, the 6th pipe does not add need testing solution and makes the blank pipe, the 7th pipe does not still add need testing solution and replaces normal saline with distilled water, as hemolytic positive control.Send in 37 ℃ of constant water bath box after each pipe shakes up gently and hatch, observe once respectively at 0.25h, 0.5h, 1h, 2h, 3h, 4h, continuous 6 times, as the clear and bright redness of solution, it is residual that the pipe end is acellular is full haemolysis; The clear and bright redness of solution or brown, it is part haemolysis that the pipe end has a small amount of erythrocyte residual; As brownish red or rufous flocculent deposit are arranged in the solution, can not disperse after the jolting, expression has erythroagglutination; All sink as erythrocyte, upper strata liquid achromatism and clarity can be judged as no haemolysis.
(3) result of the test
As seen from Table 1, concentration is every milliliter of medicine of the present invention that contains the 2mg troxerutin and encephalic protein hydrolysate, 0.1,0.2,0.3,0.3,04, the 0.5ml pipe, and 37 ℃ of insulation 4h, not seeing has haemolysis and agglutination.Upper strata liquid water white transparency, erythrocyte all sinks, and shows that every milliliter contains 2mg troxerutin and encephalic protein hydrolysate and no obvious haemolysis, meets the requirement of used for intravenous injection security inspection.
Table 1 troxerutin and encephalic protein hydrolysate solution hemolytic result of the test
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | |
| Troxerutin and encephalic protein hydrolysate (m1) normal saline (ml) 2% red blood cell suspension (ml) result | 0.1 2.4 2.5 - | 0.2 2.3 2.5 - | 0.3 2.2 2.5 - | 0.4 2.1 2.5 - | 0.5 2.0 2.5 - | - 2.5 2.5 - | -distilled water 2.5+ |
Annotate: "+" expression haemolysis among the result; "-" represents not haemolysis and do not have red cell agglutination.
Two. irritation test
(1) rabbit auricular vein irritant test
Get 3 of rabbit, every day every rabbit one pick up the ears the edge vein by sterile working's method respectively the next day instillation concentration be every milliliter of solution 20ml/kg that contains troxerutin and encephalic protein hydrolysate 2mg, once a day, side isopyknic normal saline that instils in addition, continuous five times, 24h perusal instillation position has or not congested detumescence and tissue necrosis phenomenon and sacrificed by exsanguination after the last administration, entad hold 1.5～3cm place to separate blood vessel at the distance injection point, the routine paraffin wax embedding of drawing materials, section, make the pathology tissue examination after the H.E dyeing, observation has or not vasodilation, and tube wall destroys, endotheliocytic swelling, intracavity thrombosis, significance such as tissue degeneratiaon or necrosis irritant reaction.
Result of the test shows does not see obvious hyperemia and edema, and vessel boundary is clear, and pathological changes such as tissue degeneratiaon and necrosis (seeing Fig. 1, tame rabbit ear edge blood vessel irritation test pathological section) are not seen in the pathological tissue inspection.
(2) rabbit quadriceps femoris irritant test
Injection of troxerutin and encephalic protein hydrolysate is respectively got 2 of rabbit, in a side quadriceps femoris meat by sterile working's method respectively implantation concentration be the injection of troxerutin and encephalic protein hydrolysate 1ml of 32mg/ml, the isopyknic sterile water for injection of side injection in addition, sacrificed by exsanguination behind the 48h, observation is tried the irritant reaction of thing, be converted into corresponding reaction score value by table 2, to calculate the summation of 4 quadriceps femoris reaction score values.
Table 2 muscular irritation reaction standards of grading
| The reaction score value | Irritant reaction |
| 0 1 2 3 4 5 | No significant reaction mild hyperaemia, diameter is in the following moderate hyperemia of 0.5cm, diameter is in the following severe hyperemia of 1.0cm, with the degeneration of myodegeneration muscle brown, the serious degeneration of downright bad muscle occurs, occur large stretch of downright bad |
Result of the test shows that every quadriceps femoris of rabbit there is no the obvious stimulation reaction; 4 quadriceps femoris reaction score values are 0, and pathological changes (seeing Fig. 2, rabbit quadriceps femoris irritation test pathological section) such as tissue degeneratiaon and necrosis are not seen in the pathological tissue inspection, show that the present invention is subjected to reagent that the rabbit quadriceps femoris is not had the obvious stimulation effect.
Three. whole body causes hypersensitive test
(1) experimental animal: healthy albino guinea-pig, male and female dual-purpose (female Cavia porcellus does not have pregnant), body weight 230～300g is provided by Animal Experimental Study center, Hubei Province.Credit number: SCXK (Hubei Province) 2003-0005.
(2) be subjected to the reagent thing: injection of troxerutin and encephalic protein hydrolysate, specification 80mg: 1mg (troxerutin: total nitrogen), lot number: 20030701.Provide by the gloomy medical biotechnology company limited of Hainan Hewlett-Packard.Refrigerator is preserved, and faces that (Hunan Cologne Pharmaceutical Co., Ltd produces, lot number: A040903-01) dissolve, and be diluted to every milliliter and contain the 2mg troxerutin and encephalic protein hydrolysate as sensitization liquid with preceding usefulness 5% glucose injection.
(3) be subjected to egg on probation: available from market, take out Ovum Gallus domesticus album by the sterile working during use, be made into 1% concentration with distilled water.
(4) test method
18 of extracting waste Cavia porcelluss are divided into normal saline group, 1% fresh albumen group, troxerutin and encephalic protein hydrolysate group of the present invention at random by sex, body weight, every group of 6 animals.3 treated animals difference every other day intraperitoneal injection of saline, 1% fresh albumen, troxerutin and encephalic protein hydrolysate solution of the present invention, 0.5ml/, totally 3 times.Then, every group get respectively 3 after the 1st administration the 14th day and the 21st day, auricular vein injecting normal saline, 1% fresh albumen, troxerutin and encephalic protein hydrolysate solution of the present invention, 1.0ml/ are only.Observe Cavia porcellus injects whether perpendicular hair is arranged in back 15 minutes, phenomenons such as excited uneasiness, dyspnea, sneeze, retch or cough, as two or more symptom occurs, or one of tic, collapse, phenomena of mortality person arranged, be considered as the anaphylaxis positive.
(5) result of the test
Behind normal saline or troxerutin and encephalic protein hydrolysate solution lumbar injection of the present invention the 14th day and the 21st day, difference auricular vein injecting normal saline or troxerutin and encephalic protein hydrolysate solution of the present invention, inject in back 15 minutes, symptoms of allergic such as perpendicular hair, excited uneasiness, dyspnea, sneeze, cough, retch, tic, collapse, death do not appear in Cavia porcellus.
Behind 1% fresh albumen lumbar injection the 14th day, auricular vein was injected 1% fresh albumen, injected back 3 Cavia porcelluss and all showed excited uneasiness, dyspnea, sneeze, wherein 2 tics, death in 8 minutes.Behind 1% fresh albumen lumbar injection 21 days, auricular vein was injected 1% fresh albumen, injected back 3 Cavia porcelluss and all showed excited uneasiness, dyspnea, sneeze, tic, all death in 3 minutes.
Conclusion (of pressure testing)
Concentration is that every milliliter of troxerutin and encephalic protein hydrolysate that contains 2mg is not seen obvious haemolysis, and its result meets intravenous injection medication security requirement.
Rabbit auricular vein every day instillation concentration is every milliliter of troxerutin and encephalic protein hydrolysate 20ml/kg that contains 2mg, and continuous 5 times, 24h after the last administration does not see the obvious stimulation reaction to tame rabbit ear vein; Rabbit quadriceps femoris intramuscular injection concentration is that every milliliter of 32mg contains troxerutin and encephalic protein hydrolysate 1ml, does not see the reaction of rabbit quadriceps femoris obvious stimulation, shows that the local excitation test of troxerutin and encephalic protein hydrolysate of the present invention meets the injecting drug use regulation.
Albino guinea-pig lumbar injection concentration is every milliliter of troxerutin and encephalic protein hydrolysate that contains 2mg three times, carry out 14 and after 21d attacks respectively from vein, do not see that animal grabs systemic anaphylaxis such as nose, cough, amyostasia and perpendicular hair, the reaction score value is zero, shows that troxerutin and encephalic protein hydrolysate does not have the anaphylaxis of causing to Cavia porcellus.
Claims (3)
1. injection of troxerutin and encephalic protein hydrolysate, it is characterized in that forming by troxerutin and Cerebrolysin Vial, the two weight proportion is: troxerutin: Cerebrolysin Vial=50～100: 1～20, wherein Cerebrolysin Vial content is by the free amino acid total nitrogen, the pH value 5.5～7.5 of compositions.
2. injection of troxerutin and encephalic protein hydrolysate according to claim 1 is characterized in that troxerutin: Cerebrolysin Vial=80: 1, wherein Cerebrolysin Vial content is by free amino acid total nitrogen Cerebrolysin Vial total nitrogen.
3. the preparation method of the injection of troxerutin and encephalic protein hydrolysate of a claim 1, it is characterized in that troxerutin and Cerebrolysin Vial stock solution are mixed in proportion, add an amount of water for injection to prescribed volume, press amount of liquid 0.07% and add the injection stage medicinal charcoal, stirring and adsorbing is 30 minutes under the room temperature, earlier through coarse filtration, then through the microporous filter membrane fine straining of 0.22 μ m, pack into filtrate branch in the cillin bottle, put (pre-freeze is to-40 ℃) in the freeze drying box, insulation is 2 hours under this temperature; Start the water vessel switch, start vacuum pump, under 4Pa, gradually with per hour 1 ℃) improve temperature, when arriving room temperature, continued evacuation 3 hours, lyophilizing finishes, sterile packaged.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104888184A (en) * | 2015-05-19 | 2015-09-09 | 邓学峰 | A pharmaceutical composition of cerebroprotein hydrolysate and applications thereof |
| CN113908174A (en) * | 2021-10-29 | 2022-01-11 | 北京四环制药有限公司 | Efficient and safe preparation method and application of cerebroprotein hydrolysate |
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2005
- 2005-08-26 CN CN 200510094044 patent/CN1736475A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104888184A (en) * | 2015-05-19 | 2015-09-09 | 邓学峰 | A pharmaceutical composition of cerebroprotein hydrolysate and applications thereof |
| CN113908174A (en) * | 2021-10-29 | 2022-01-11 | 北京四环制药有限公司 | Efficient and safe preparation method and application of cerebroprotein hydrolysate |
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