CN104940914A - New application of human urine kininogenase and medicine composition comprising same - Google Patents
New application of human urine kininogenase and medicine composition comprising same Download PDFInfo
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- CN104940914A CN104940914A CN201510403851.5A CN201510403851A CN104940914A CN 104940914 A CN104940914 A CN 104940914A CN 201510403851 A CN201510403851 A CN 201510403851A CN 104940914 A CN104940914 A CN 104940914A
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Abstract
The invention belongs to the field of medicine, and particularly relates to a new application of human urine kininogenase and a medicine composition comprising the human urine kininogenase. The human urine kininogenase is applied to prepare medicine for treating the Alzheimer's disease. The invention further provides the medicine composition for treating the Alzheimer's disease. The medicine composition comprises the human urine kininogenase and bilobalide, and further comprises pentoxifyline. The medicine composition can remarkably recover the memory capacity of a patient suffering with the Alzheimer's disease, improve the cognition capacity of the patient suffering with the Alzheimer's disease, delay the disease progress of the Alzheimer's disease, greatly improve the life quality of the patient suffering with the Alzheimer's disease, improve the treatment compliance of the Alzheimer's disease, improve the treatment effect, provide the effective treatment medicine for the patient suffering with the Alzheimer's disease, and greatly relieve pain of the patient of the Alzheimer's disease.
Description
Technical field
The invention belongs to field of medicaments, be specifically related to the novelty teabag of Human Urinary Kallidinogenase and the pharmaceutical composition containing Human Urinary Kallidinogenase.
Background technology
Human Urinary Kallidinogenase (Human Urinary Kallidinogenase) is a kind of serine protease be made up of 238 aminoacid extracted from human urine, its Main Function is in vivo that kininogen is hydrolyzed to kassinin kinin, thus exercise a series of physiological function, having expansion blood capillary, lax vascular smooth muscle, improve microcirculation, increase the effects such as blood flow, anticoagulant, thrombus dissolving, reduction blood viscosity and blood pressure lowering, is a kind of protease with higher medical value.
In recent years, along with people are to the further investigation of Human Urinary Kallidinogenase, find the increasing novelty teabag of Human Urinary Kallidinogenase.Such as, Chinese patent 101134106B discloses a kind of pharmaceutical composition containing Human Urinary Kallidinogenase being used for the treatment of cerebral infarction, and Chinese patent CN101879308B discloses Human Urinary Kallidinogenase's application in preparation treatment renal failure medicine etc.Therefore, Human Urinary Kallidinogenase has wide medical prospect.
Senile dementia is the degenerative disease of a kind of central nervous system.Its clinical manifestation is continuous deterioration that is cognitive and memory ability, and activity of daily living Progressive symmetric erythrokeratodermia goes down and occurs various all schizophrenic symptoms and behavior disorder.The life threatening old people that senile dementia is serious, brings white elephant to family and society.Thus, research and develop out a kind of medicine effectively can treating senile dementia and there is very important social meaning and clinical meaning.
At present, the research and development for the treatment of senile dementia cause the great attention of countries in the world the world of medicine, along with scientists is to the further investigation of old nervous physiology, biochemistry, pharmacology aspect, the drug development of associated treatment senile dementia also achieves larger progress.Now, the medicine for the treatment of senile dementia mainly contains cholinesterase inhibitor, cerebral vasodilator, calcium antagonists, prevents A β from depositing medicine, suppresses β, gamma secretase medicine and anti-inflammatory agent and have the natural component of potential anti-senile dementia effect.Although said medicine has demonstrated certain curative effect in clinical practice, in life-time service process, there is serious inevitable side effect, and therapeutic effect is not remarkable.Therefore, finding a kind of medicine for the treatment of senile dementia Be very effective, side effect little is the difficult problem needing solution at present badly.
Bilobalide is the natural product extracted from Gingko biloba plant, is a class specificity platelet activating factor antagonist.Bilobalide has restitution to cholinergic impair memory function, can alleviate and improve myocardial ischemia simultaneously and have effect antianxity.And bilobalide can also be used for the treatment of thrombosis, acute pancreatitis and cardiovascular disease clinically, also has antioxidation simultaneously, the effect of slow down aging is the natural drug that a kind of medical value is very high.
Pentoxifylline is the derivant of methylxanthine, is a kind of non-selective phosphodiesterase inhibitor, is commonly used for blood vessel dilating, anticoagulant, the hemorheological effect of improvement for the treatment of peripheral blood vessel, cardiovascular and cerebrovascular disease.Hu Rui etc. have delivered at " Nanfang Medical Univ's journal " paper that a section is entitled as " experimentation that pentoxifylline promotes learning and memory effect ", this paper shows that pentoxifylline can improve the Spatial learning ability of senile dementia rat, is a kind of medicine with improving studing ability.
Summary of the invention
The invention provides the novelty teabag of Human Urinary Kallidinogenase and the pharmaceutical composition containing Human Urinary Kallidinogenase, be specifically related to the purposes of Human Urinary Kallidinogenase in preparation treatment senile dementia, additionally provide the pharmaceutical composition containing Human Urinary Kallidinogenase simultaneously.
The invention provides the purposes of Human Urinary Kallidinogenase in preparation treatment senile dementia.
Further, described Human Urinary Kallidinogenase is injection, and described injection is also containing mannitol, sodium chloride, glucose or Polyethylene Glycol.
Further, present invention also offers a kind of pharmaceutical composition for the treatment of senile dementia, described pharmaceutical composition comprises Human Urinary Kallidinogenase and bilobalide.
Further, in described pharmaceutical composition, the mass ratio of Human Urinary Kallidinogenase and bilobalide is 1:0.1-1, and the mass ratio of described Human Urinary Kallidinogenase and bilobalide is preferably 1:0.4.
Further, described pharmaceutical composition also comprises pentoxifylline.
Further, in described pharmaceutical composition, the weight ratio of Human Urinary Kallidinogenase and pentoxifylline is 1:0.05-0.4, and the weight ratio of described Human Urinary Kallidinogenase and pentoxifylline is preferably 1:0.25.
The pharmaceutical composition for the treatment of senile dementia provided by the invention is injection, and described injection is also containing mannitol, sodium chloride, glucose or Polyethylene Glycol.
Senile dementia is the degenerative disease of a kind of central nervous system.Its clinical manifestation is cognitive and memory ability decline.The present invention selects APP/PS1 bi-transgenic mice to be the animal model of senile dementia, keeps away dark latency as the index of learning memory measuring APP/PS1 bi-transgenic mice using what keep away that dark test behavioristics detects APP/PS1 bi-transgenic mice.Inventor proves through test, what Human Urinary Kallidinogenase significantly can shorten APP/PS1 bi-transgenic mice keeps away dark latency, improve the learning memory of APP/PS1 bi-transgenic mice, to the senile dementia of APP/PS1 bi-transgenic mice, there is certain therapeutic effect.
Bilobalide is the natural product extracted from Gingko biloba plant, is a class specificity platelet activating factor antagonist.Bilobalide can be used for the ischemic cerebrovascular that stagnation of QI and blood causes, and pentoxifylline is a kind of medicine with improving studing ability.Inventor finds through test, what the pharmaceutical composition of Human Urinary Kallidinogenase and bilobalide significantly can shorten APP/PS1 bi-transgenic mice keeps away dark latency, than the Be very effective of Human Urinary Kallidinogenase and the independent medication of bilobalide, illustrate that Human Urinary Kallidinogenase and bilobalide have good synergism shortening keeping away in dark latency of APP/PS1 bi-transgenic mice.Simultaneously, what urinary kallidinogenase, bilobalide and pentoxifylline pharmaceutical composition extremely significantly can shorten APP/PS1 bi-transgenic mice keeps away dark latency, its effect is than the Be very effective of the pharmaceutical composition of Human Urinary Kallidinogenase and bilobalide, illustrate that urinary kallidinogenase, bilobalide and pentoxifylline have significant synergism keeping away in dark latency of shortening APP/PS1 bi-transgenic mice, the ability of learning and memory of APP/PS1 bi-transgenic mice can be significantly improved, to senile dementia, there is good therapeutic effect.
In addition, the present invention detects the resolving index of APP/PS1 bi-transgenic mice as the cognitive competence measuring APP/PS1 bi-transgenic mice using new object recognition test.Inventor finds through test, and urinary kallidinogenase can improve the resolving index of APP/PS1 bi-transgenic mice, improves the cognitive competence of APP/PS1 bi-transgenic mice.Human Urinary Kallidinogenase compares with the independent medication of bilobalide with Human Urinary Kallidinogenase with the pharmaceutical composition of bilobalide, this pharmaceutical composition can improve the resolving index of APP/PS1 bi-transgenic mice significantly, illustrates that Human Urinary Kallidinogenase and the resolving index that bilobalide is improving APP/PS1 bi-transgenic mice have good synergism.Simultaneously, urinary kallidinogenase, bilobalide and pentoxifylline pharmaceutical composition can improve the resolving index of APP/PS1 bi-transgenic mice extremely significantly, its effect is than the Be very effective of the pharmaceutical composition of Human Urinary Kallidinogenase and bilobalide, illustrate that urinary kallidinogenase, bilobalide and pentoxifylline have significant therapeutic effect in the resolving index improving APP/PS1 bi-transgenic mice, to senile dementia, there is good therapeutic effect.
In a word, compared with prior art, technical scheme provided by the invention has the following advantages:
(1) purposes of Human Urinary Kallidinogenase provided by the invention in preparation treatment senile dementia, Human Urinary Kallidinogenase can delay the Development process of senile dementia, for patients of senile dementia provides a kind of effective medicine.
(2) pharmaceutical composition for the treatment of senile dementia provided by the invention, this pharmaceutical composition comprises Human Urinary Kallidinogenase and bilobalide, described pharmaceutical composition can recover the memory ability of patients of senile dementia significantly, can also the effective cognitive competence improving patients of senile dementia.
(3) pharmaceutical composition for the treatment of senile dementia provided by the invention, this pharmaceutical composition comprises Human Urinary Kallidinogenase and bilobalide, also comprise pentoxifylline, described pharmaceutical composition effect in the cognitive competence of the memory ability and raising patients of senile dementia that recover patients of senile dementia is more remarkable than the pharmaceutical composition of Human Urinary Kallidinogenase and bilobalide, greatly can improve the quality of life of patients of senile dementia, effectively alleviate the misery of patients of senile dementia.
detailed description of the invention:
Below by way of the description of detailed description of the invention, the invention will be further described, but this is not limitation of the present invention, those skilled in the art are according to basic thought of the present invention, various amendment or improvement can be made, but only otherwise depart from basic thought of the present invention, all within the scope of the present invention.
embodiment 1,human Urinary Kallidinogenase's injection
Get the Human Urinary Kallidinogenase 3 milligrams of filtration sterilization, add appropriate water for injection and dissolve, regulate pH to neutral, inject water to 500 milliliters, add sodium chloride and regulate isotonic, aseptic filtration, in subpackage 1000 ampoules, to obtain final product.
embodiment 2,the injection of Human Urinary Kallidinogenase and bilobalide pharmaceutical composition
Get the Human Urinary Kallidinogenase 2 milligrams of filtration sterilization, add appropriate water for injection and dissolve, then add 0.2 milligram of bilobalide dissolving, regulate pH to neutral, inject water to 500 milliliters, add sodium chloride and regulate isotonic, aseptic filtration, in subpackage 1000 ampoules, to obtain final product.
embodiment 3,the injection of Human Urinary Kallidinogenase and bilobalide pharmaceutical composition
Get the Human Urinary Kallidinogenase 3 milligrams of filtration sterilization, add appropriate water for injection and dissolve, then add 1.2 milligrams of bilobalides dissolvings, regulate pH to neutral, inject water to 500 milliliters, add sodium chloride and regulate isotonic, aseptic filtration, in subpackage 1000 ampoules, to obtain final product.
embodiment 4,the injection of Human Urinary Kallidinogenase and bilobalide pharmaceutical composition
Get the Human Urinary Kallidinogenase 3.5 milligrams of filtration sterilization, add appropriate water for injection and dissolve, then add 2.8 milligrams of bilobalides dissolvings, regulate pH to neutral, inject water to 500 milliliters, add Polyethylene Glycol and regulate isotonic, aseptic filtration, in subpackage 1000 ampoules, to obtain final product.
embodiment 5,the injection of Human Urinary Kallidinogenase, bilobalide and pentoxifylline pharmaceutical composition
Get the Human Urinary Kallidinogenase 3.5 milligrams of filtration sterilization, add appropriate water for injection to dissolve, add 1.4 milligrams of bilobalides and 0.175 milligram of pentoxifylline alkali dissolution again, regulate pH to neutral, inject water to 500 milliliters, add sodium chloride and regulate isotonic, aseptic filtration, in subpackage 1000 ampoules, to obtain final product.
embodiment 6,the injection of Human Urinary Kallidinogenase, bilobalide and pentoxifylline pharmaceutical composition
Get the Human Urinary Kallidinogenase 3 milligrams of filtration sterilization, add appropriate water for injection to dissolve, add 1.2 milligrams of bilobalides and 0.75 milligram of pentoxifylline alkali dissolution again, regulate pH to neutral, inject water to 500 milliliters, add sodium chloride and regulate isotonic, aseptic filtration, in subpackage 1000 ampoules, to obtain final product.
test example one,human Urinary Kallidinogenase tests the impact of the memory ability of APP/PS1 bi-transgenic mice
1, subjects: choose 70 APP/PS1 bi-transgenic mice, provided by Nanjing Medical University's Experimental Animal Center.
2, test grouping: get 70 APP/PS1 bi-transgenic mice and be divided into 7 groups at random, often organize 10, be respectively model group, Human Urinary Kallidinogenase's low dose group, Human Urinary Kallidinogenase's high dose group, bilobalide group, pentoxifylline group, Human Urinary Kallidinogenase+bilobalide group, Human Urinary Kallidinogenase+bilobalide+pentoxifylline group.The administering mode of each group of rat is rear tail vein injection, and once a day, continuous 4 weeks, each grouping dosage was as follows:
Model group: inject isopyknic normal saline;
Human Urinary Kallidinogenase's low dose group: Human Urinary Kallidinogenase's injection of injection 0.1mg/kg;
Human Urinary Kallidinogenase's high dose group: Human Urinary Kallidinogenase's injection of injection 0.5mg/kg;
Bilobalide group: the bilobalide injection of injection 0.2mg/kg;
Pentoxifylline group: the pentoxifylline injection of injection 0.125mg/kg;
Human Urinary Kallidinogenase+bilobalide group: injection Human Urinary Kallidinogenase 0.5mg/kg+ bilobalide 0.2mg/kg;
Human Urinary Kallidinogenase+bilobalide+pentoxifylline group: injection Human Urinary Kallidinogenase 0.5mg/kg+ bilobalide 0.2mg/kg+ pentoxifylline 0.125mg/kg.
3, test method:
Employing keeps away dark auto testing instrument and detects, and keeps away active box point light and shade two Room of dark auto testing instrument, has a hole, pass to copper grid at the bottom of case between two Room.Before formal test, each group of APP/PS1 bi-transgenic mice is trained, APP/PS1 bi-transgenic mice head is carried hole and puts into bright room, first conform 2min, lead to 36V electric current then to darkroom copper grid, APP/PS1 bi-transgenic mice is subject to electric shock and namely runs away to bright room after entering darkroom, copper grid continue energising 5min, as training process.The test of memory of APP/PS1 bi-transgenic mice is carried out after 24h, record APP/PS1 bi-transgenic mice enters the time (keeping away dark incubation period) in darkroom for the first time, if enter darkroom not yet in APP/PS1 bi-transgenic mice 5min, its incubation period, 300s made by meter.4, result of the test
Result of the test is as shown in table 1.
Table 1 each administration group keeps away dark preclinical impact (± SD) to APP/PS1 bi-transgenic mice
Compared with model group, * P < 0.05, * * P < 0.01;
Compared with Human Urinary Kallidinogenase's high dose group,
★p < 0.05,
★ ★p < 0.01;
Compared with Human Urinary Kallidinogenase+bilobalide group,
■p < 0.05,
■ ■p < 0.01.
As can be seen from Table 1:
(1) compared with model group, what Human Urinary Kallidinogenase significantly can shorten APP/PS1 bi-transgenic mice keeps away dark latency, and especially the keep away dark latency of Human Urinary Kallidinogenase's high dose group to APP/PS1 bi-transgenic mice shortens and have extremely significant difference (p < 0.01);
(2) compared with Human Urinary Kallidinogenase's high dose group, what the pharmaceutical composition of Human Urinary Kallidinogenase and bilobalide can shorten APP/PS1 bi-transgenic mice significantly keeps away dark latency, illustrates that Human Urinary Kallidinogenase and the keeping away in dark latency of APP/PS1 bi-transgenic mice that bilobalide is shortening have good synergism;
(3) compared with Human Urinary Kallidinogenase+bilobalide group, what the pharmaceutical composition of Human Urinary Kallidinogenase, bilobalide and pentoxifylline can shorten APP/PS1 bi-transgenic mice extremely significantly keeps away dark latency, illustrate that Human Urinary Kallidinogenase, bilobalide and pentoxifylline have significant synergism keeping away in dark latency of shortening APP/PS1 bi-transgenic mice, significantly can recover the memory ability of APP/PS1 bi-transgenic mice.
test example two,human Urinary Kallidinogenase tests the impact of the cognitive competence of APP/PS1 bi-transgenic mice
1, subjects: choose 70 APP/PS1 bi-transgenic mice, provided by Nanjing Medical University's Experimental Animal Center.
2, test grouping: get 70 APP/PS1 bi-transgenic mice and be divided into 7 groups at random, often organize 10, be respectively model group, Human Urinary Kallidinogenase's low dose group, Human Urinary Kallidinogenase's high dose group, bilobalide group, pentoxifylline group, Human Urinary Kallidinogenase+bilobalide group, Human Urinary Kallidinogenase+bilobalide+pentoxifylline group.The administering mode of each group of rat is rear tail vein injection, and once a day, continuous 4 weeks, each grouping dosage was as follows:
Model group: inject isopyknic normal saline;
Human Urinary Kallidinogenase's low dose group: Human Urinary Kallidinogenase's injection of injection 0.1mg/kg;
Human Urinary Kallidinogenase's high dose group: Human Urinary Kallidinogenase's injection of injection 0.5mg/kg;
Bilobalide group: the bilobalide injection of injection 0.2mg/kg;
Pentoxifylline group: the pentoxifylline injection of injection 0.125mg/kg;
Human Urinary Kallidinogenase+bilobalide group: injection Human Urinary Kallidinogenase 0.5mg/kg+ bilobalide 0.2mg/kg;
Human Urinary Kallidinogenase+bilobalide+pentoxifylline group: injection Human Urinary Kallidinogenase 0.5mg/kg+ bilobalide 0.2mg/kg+ pentoxifylline 0.125mg/kg.
3, test method:
Adopt new object identification method to test, test and carry out in a homemade white plastic behavior case, the length, width and height of behavior case are respectively 50*50*30cm, and whole test divides 3 days 3 stages to complete.First stage, the 1st day is the laundering period, mice is put into behavior case and allows it freely explore 5 minutes, at this one-phase, do not have data collection.Second stage, the 2nd day is training period, and two identical objects 1 and object 2 are put into behavior case, the about 5cm of object distance behavior case, and two objects, at a distance of 20cm, are put into mice and allowed it freely explore 5 minutes.Exploratory behaviour is defined as: the nose distance object of mice is less than 2cm or touches object with nose, and walking about around object or lie prone can not as exploratory behaviour at its near vicinity.At this one-phase, record mice explores the time of each object, and after terminating, mice puts back to cage at once.Phase III, the 3rd day is the testing period, and the behavior case be placed on by mice after object 1 being changed into a new object 3 freely explores 5 minutes.Resolving index is adopted to weigh the cognitive function of mice, the new object of resolving index=explore time/(explore new object time+time of exploration past heritage body), namely training period resolving index=exploration object 1 time/(explore object 1 time+time of explorations object 2), the resolving index=exploration object 3 of testing period time/(exploration object 3 time+time of exploration object 2).
4, result of the test
Result of the test is as shown in table 2.
The resolving index of table 2 APP/PS1 bi-transgenic mice training period and testing period
Compared with model group, * P < 0.05, * * P < 0.01;
Compared with Human Urinary Kallidinogenase's high dose group,
★p < 0.05,
★ ★p < 0.01;
Compared with Human Urinary Kallidinogenase+bilobalide group,
■p < 0.05,
■ ■p < 0.01.
As shown in Table 2:
(1) respectively organized for training period the time that mice explores two same object to add up, find that each group of mice explores the resolving index no difference of science of statistics of two same object.
(2) respectively organized for the testing period time that mice explores two different objects to add up:
(A) compared with model group, Human Urinary Kallidinogenase can significantly improve APP/PS1 bi-transgenic mice resolving index, and especially the raising of Human Urinary Kallidinogenase's high dose group to APP/PS1 bi-transgenic mice resolving index has extremely significant difference (p < 0.01);
(B) compared with Human Urinary Kallidinogenase's high dose group, the pharmaceutical composition of Human Urinary Kallidinogenase and bilobalide can improve the resolving index of APP/PS1 bi-transgenic mice significantly, illustrates that Human Urinary Kallidinogenase and bilobalide have good synergism in the resolving index improving APP/PS1 bi-transgenic mice;
(C) Human Urinary Kallidinogenase+bilobalide group is compared, the pharmaceutical composition of Human Urinary Kallidinogenase, bilobalide and pentoxifylline can improve the resolving index of APP/PS1 bi-transgenic mice extremely significantly, illustrate that Human Urinary Kallidinogenase, bilobalide and pentoxifylline have significant synergism in the resolving index improving APP/PS1 bi-transgenic mice, the cognitive competence of APP/PS1 bi-transgenic mice can be improved significantly.
Claims (9)
1. the purposes of Human Urinary Kallidinogenase in preparation treatment senile dementia.
2. purposes as claimed in claim 1, it is characterized in that, described Human Urinary Kallidinogenase is injection, and described injection is also containing mannitol, sodium chloride, glucose or Polyethylene Glycol.
3. treat a pharmaceutical composition for senile dementia, it is characterized in that, described pharmaceutical composition comprises Human Urinary Kallidinogenase and bilobalide.
4. the pharmaceutical composition for the treatment of senile dementia as claimed in claim 3, it is characterized in that, in described pharmaceutical composition, the mass ratio of Human Urinary Kallidinogenase and bilobalide is 1:0.1-1.
5. the pharmaceutical composition for the treatment of senile dementia as claimed in claim 4, it is characterized in that, in described pharmaceutical composition, the mass ratio of Human Urinary Kallidinogenase and bilobalide is 1:0.4.
6. the pharmaceutical composition for the treatment of senile dementia as claimed in claim 3, it is characterized in that, described pharmaceutical composition also comprises pentoxifylline.
7. the pharmaceutical composition for the treatment of senile dementia as claimed in claim 6, it is characterized in that, in described pharmaceutical composition, the weight ratio of Human Urinary Kallidinogenase and pentoxifylline is 1:0.05-0.4.
8. the pharmaceutical composition for the treatment of senile dementia as claimed in claim 7, it is characterized in that, in described pharmaceutical composition, the weight ratio of Human Urinary Kallidinogenase and pentoxifylline is 1:0.25.
9. the pharmaceutical composition of the treatment senile dementia as described in as arbitrary in claim 3-8, it is characterized in that, described pharmaceutical composition is injection, and described injection is also containing mannitol, sodium chloride, glucose or Polyethylene Glycol.
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| CN106955351A (en) * | 2016-01-08 | 2017-07-18 | 周艳玲 | A kind of pharmaceutical composition of anti-senile dementia |
| CN109091667A (en) * | 2018-09-18 | 2018-12-28 | 广东天普生化医药股份有限公司 | Purposes and combinations thereof of the human urokinase-type peptidase in preparation treatment migraine remedy |
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| CN101879308A (en) * | 2010-06-13 | 2010-11-10 | 广东天普生化医药股份有限公司 | Application of human urinary kallidinogenase in preparing medicine for treating acute renal failureacute renal failure |
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| CN101879308A (en) * | 2010-06-13 | 2010-11-10 | 广东天普生化医药股份有限公司 | Application of human urinary kallidinogenase in preparing medicine for treating acute renal failureacute renal failure |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106955351A (en) * | 2016-01-08 | 2017-07-18 | 周艳玲 | A kind of pharmaceutical composition of anti-senile dementia |
| CN109091667A (en) * | 2018-09-18 | 2018-12-28 | 广东天普生化医药股份有限公司 | Purposes and combinations thereof of the human urokinase-type peptidase in preparation treatment migraine remedy |
| CN109091667B (en) * | 2018-09-18 | 2020-05-05 | 广东天普生化医药股份有限公司 | Application of human urinary kallidinogenase in preparing medicine for treating migraine and composition thereof |
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