CN103191142A - Baicalin and taurine composition and application thereof in preparation of medicament - Google Patents

Baicalin and taurine composition and application thereof in preparation of medicament Download PDF

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CN103191142A
CN103191142A CN2013100975613A CN201310097561A CN103191142A CN 103191142 A CN103191142 A CN 103191142A CN 2013100975613 A CN2013100975613 A CN 2013100975613A CN 201310097561 A CN201310097561 A CN 201310097561A CN 103191142 A CN103191142 A CN 103191142A
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baicalin
taurine
dementia
compositions
group
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CN103191142B (en
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杨小玲
张占军
谢宁
吕武清
刘地发
程帆
王章伟
欧阳婷
蒋春红
王燕平
崔刚
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JIANGXI QINGFENG PHARMACEUTICAL CO Ltd
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Abstract

The invention relates to a baicalin and taurine medicinal composition capable of protecting neurocyte injury, improving the learning and memory ability and improving the cognition impairment so as to prevent and treat dementia.

Description

Baicalin and taurine composition and preparation medicinal usage thereof
Technical field
The present invention relates to baicalin and taurine composition and for the preparation of the purposes of medicine.
Background technology
Dementia is normal sending out in a kind of central nervous system degenerative disease of geratic period and presenium, be that, dysmnesia damaged with cognitive function, the hypophrenia, personality changes and dystropy are one group of syndrome of principal character, cause patient's occupation and social life function reduction or forfeiture until death at last.In recent years, along with increase year by year and the aggravation of aging problem of world population average life, its sickness rate presents the trend that rises year by year, and human life quality's influence and even the threat of existence are also become clear day by day.The mean survival time (MST) of dementia patients only is 5~6 years, has become to be only second to the fourth-largest disease that causes old people's death after the cardiovascular disease, cancer, cerebrovascular disease.
According to etiology, clinical manifestation, sick damage situation etc. can be divided into different dull-witted hypotypes: Alzheimer (AD), vascular dementia (VaD), the mixed type of Alzheimer and vascular dementia (MIXD), dementia with Lewy body (DLB), parkinson dementia (PD), volume temporal lobe dementia (FTD), dementia in Creutzfeldt-Jakob disease (CJD), normal pressure hydrocephalus dementia (NPH), Wei-Ke two formula syndrome dementias (WKS), the dementia that Huntington Chorea is relevant waits other pathological changes.Wherein AD is topmost dull-witted hypotype, accounts for 60%~70% of all dementias, is VaD or MIXD secondly.
In the nearly many decades, all obtained significant progress at basis and the clinical research of dementia disease, and mainly be the research to most common dementia hypotype AD.Nonetheless, because of the cause of disease of dementia and the complexity of pathomechanism, still there are various disputes, the various theories whiles also deposit, as " A β theory ", " neuron loss theory ", " neurofibrillary tangles theory (as Tua abnormal protein phosphorylation) ", " the unusual theory of neurotransmitter (reducing as acetylcholine Ach) ", " toxicity of excitatory amino acid theory (as glutamic acid Glu hyperfunctioning) ", " free radical and oxidative damage theory ", " abnormal carbohydrate metabolism theory " etc., really do not refer to; Recent progress that still can't making a breakthrough property.But dementia is one group of syndrome, although can be formed by difference or Different types of etiopathogenises, but the disease of its formation has identical symptom performance, therefore in the treatment of dementia, reach common understanding, namely the dull-witted main purpose for the treatment of is to improve symptom, comprises raising cognitive competence and self-care ability, the neural mental act obstacle of improvement; Change progression of disease, delay or stop the severity of symptoms of dull-witted process or intervene the primary prevention of crucial mechanism of causing a disease.
According to effectiveness and data of safety, existing severally get the Green Light be used to the medicine that improves the dementia of the Alzheimer type symptom, as belong to cholinesterase inhibitor donepezil (aricept), Li Fansi bright, galantamine and belong to the memantine of glutamate receptor antagonists, but it is limited that the curative effect of having established is still, and needs the associating other drug to reach therapeutic effect at a specified future date.Based on etiology and the Clinical symptoms of the uniqueness of dementia, and the research state in present dull-witted field, still be badly in need of the different medicine of research and development, or act on disease symptoms and improve, or at the modification of its different causes of disease and pathophysiological process.Therefore the treatment of dementia such as AD remains a pendent important research field, also is a urgent and thorny society and difficult medical problem.
Summary of the invention
The objective of the invention is at an above-mentioned dementia treatment difficult problem based on AD; through a large amount of experiment screening and checkings; provide a kind of can the neuroprotective cell injury, improve ability of learning and memory, improve cognitive dysfunction, thereby prevention and the dull-witted pharmaceutical composition for the treatment of.
On the one hand, the invention provides baicalin and taurine composition for the preparation of the purposes of neuroprotective cell injury medicine.
On the other hand, the present invention also provides baicalin and taurine composition for the preparation of the purposes of improving the ability of learning and memory medicine.
On the other hand, the present invention also provides baicalin and taurine composition for the preparation of the purposes of improving the cognitive dysfunction medicine.
Preferably, the present invention also provides baicalin and taurine composition for the preparation of the purposes of preventing and treating dull-witted medicine.
Preferably, wherein said dementia comprises any in Alzheimer dementia (AD), vascular dementia (VD), Combination dementia (MixD), Louis corpusculum dementia (DLB), volume temporal lobe dementia (FTD), dementia in Parkinson disease (PD), dementia in Creutzfeldt-Jakob disease (CJD), normal pressure hydrocephalus dementia (NPH), the Wei-Ke two formula syndrome dementias (WKS).
Preferably, wherein said senile dementia mainly refers to Alzheimer dementia (AD), vascular dementia (VaD), Combination dementia (MixD).
Preferably, wherein said senile dementia mainly refers to Alzheimer dementia (AD).
Preferably, wherein said baicalin and taurine composition are made up of the following weight proportioning: baicalin 99.9%~60.0%, taurine 0.1~40.0%.
Preferably, wherein said baicalin and taurine composition are made up of the following weight proportioning: baicalin 99.0%~90.0%, taurine 1.0%~10.0%.
Further; the present invention also provides a kind of pharmaceutical preparation; the main effective ingredient of this pharmaceutical preparation is baicalin and taurine composition; and with one or more pharmaceutically acceptable excipient and/or can be prepared from the other medicines of its compatibility; wherein said baicalin and taurine composition are made up of the following weight proportioning: baicalin 99.9%~60.0%, taurine 0.1~40.0%.
Baicalin in baicalin of the present invention and the taurine composition is the main matter basis of Radix Scutellariae performance drug effect, has tangible antiinflammatory antiallergic action, analgesic, antibiotic, antivirus action; Can effectively remove oxygen-derived free radicals and show stronger antioxidant activity; Can raise the expression of short apoptogene p53 and bax, downward modulation apoptosis suppressor bcl-2 and bcl-6, suppress tumor cell proliferation simultaneously and have antitumor action; Suppress Ca in the cell 2+Overload and performance has arrhythmia and a hypotensive effect; Have pharmacological actions such as neuroprotective cell injury, adjusting immunologic function in addition.And what now gone on the market is preparation baicalin sheet, the baicalin capsule of main active with it, is mainly used in the auxiliary treatment of acute and chronic hepatitis and chronic persistent hepatitis.This shows that the effect between the two is different fully with therapeutic purposes.
Taurine in baicalin of the present invention and the taurine composition has antipyretic and anti-inflammatory, antalgic and sedative, hepatic cholagogic effect; Can accelerate glycolysis, strengthen glyconeogenesis and have hypoglycemic activity; Have the free radical of removing, antioxidation and suppress Ca in the cell 2+Overload effect, thereby protecting myocardial cell, delay hypertension and atherosclerotic generation and development at arrhythmia and heart failure; Have anticoagulant, promote effects such as nervous system growth promoter and cell proliferation and differentiation.What go on the market is the heating that main pharmacodynamics formulations of active ingredients taurine sheet, taurine capsule, taurine particles etc. are mainly used in alleviating common cold initial stage with the taurine; Be that the compound Siwei fructus ligustri lucidi capsule of one of drug activity composition is mainly used in menopausal syndrome and premenstrual tension syndrome with it.This shows that the effect between the two is different fully with therapeutic purposes.
Has the neuroprotective cytosis after baicalin, the taurine combination; has the effect that improves cognitive dysfunction, improves the ability of learning and memory of senile dementia; senile dementia is had the better prevention effect, and use effect is stronger separately than baicalin or taurine to make it.
Through the experimental data contrast as can be known, baicalin, taurine are separately to A β 25-35The optimum concentration that causes SH-SY5Y cell injury performance protective effect is respectively 40~80 μ mol/L, 10~80 μ mol/L.And baicalin and taurine with different proportionings combinations after, each compositions group SH-SY5Y cell relative survival rate all significantly improves than model group.Wherein the compositions that forms of the compositions, 48 μ mol/L baicalins and the 8 μ mol/L taurine proportionings that form of 40 μ mol/L baicalins and 8 μ mol/L taurine proportionings makes cell survival rate improve (90.24 ± 7.46) %, (114.39 ± 10.79) % respectively than model control group; be higher than 80 μ mol/L baicalin matched group (68.62 ± 11.54) % and 20 μ mol/L taurine matched group (83.92 ± 20.17) % in the parallel laboratory test; illustrate that compositions has stronger protective effect to neural cell injury among the present invention, be eager to excel than both independent use protective effects.
Through the experimental data contrast as can be known, rat intracerebroventricular A β 1-42After show tangible dementia symptom, the remarkable obstacle of cognitive learning ability, the escape latency of rat water maze laboratory and average search distance significantly increases than normal group, diving tower escape latency and errors number obviously increase and stop and obviously reduce incubation period.Baicalin and taurine all can obviously reduce A β with the compositions that 6: 1 proportionings, 18: 1 proportionings, 2: 1 proportionings form 1-42Cause escape latency and the average detection range of dementia rats water maze laboratory; reduce diving tower escape latency and errors number; and the diving tower that makes rat stops increase incubation period; and compositions is rapid-action; action effect obviously is better than baicalin matched group and taurine matched group, and shows certain dose-effect trend.Water maze and diving tower experiment are the classics experiments of test rat cognitive function and ability of learning and memory; baicalin and taurine composition that this description of test is formed with certain proportioning can improve cognitive function and the ability of learning and memory that A β causes dementia rats, and action effect is better than baicalin and taurine acts on separately.Point out it can be effectively to be used for improving learning and memory and cognitive dysfunction, control dementia.
Through the experimental data contrast as can be known, behind the model group KM explanation KM mouse peritoneal injection scopolamine, blocked cholinergic nerve path m receptor, the cholinergic nerve function is suppressed, escape latency and average detection range significantly increase than normal group in the mice Morris water maze laboratory, escape latency and errors number also significantly increase and stop and obviously reduce incubation period in the diving tower experiment, and mice shows significant study and work memory impairment.After giving baicalin and taurine composition, mice Morris water maze laboratory escape latency and average detection range reduce in various degree than model group, and the errors number of diving tower experiment and escape latency obviously reduce and stop and significantly increase incubation period.Baicalin and taurine reduce 52.76%, 73.53%, 26.62% with the compositions escape latency that 6: 1 proportionings, 18: 1 proportionings, 2: 1 proportionings form respectively than model group, and the average search distance reduces 39.91%, 47.98%, 24.61% respectively; Diving tower stops increases by 197.66%, 208.43%, 85.67% respectively than model group incubation period, and escape latency reduces 65.72%, 71.03%, 28.65% respectively, and errors number reduces 62.55%, 57.40%, 30.52% respectively.Experimental result shows; the compositions that baicalin and taurine proportioning form can be improved brain cholinergic nerve of centrum function; improvement causes study and work memory ability and the cognitive competence of memory acquisition disturbance mice because cholinergic nerve is impaired, the prompting baicalin can be used for improving the relevant memory impairment of cholinergic and control senile dementia with taurine composition.
New albumen synthesizes have important function in learning and memory, macromole protein and RNA synthetic closely related particularly in consolidation of memory and reproduction and the brain.The inferior amide of hexamethylene is a kind of protein synthesis inhibitor, can cause the synthetic minimizing of albumen relevant with longterm memory in the brain, has hindered the formation of memory, and has caused study and the cognitive dysfunction of brain.Therefore by consolidating obstacle modeling dementia to the inferior amide preparation of animal injection hexamethylene memory.Through the experimental data contrast as can be known, behind the inferior amide of mouse peritoneal injection hexamethylene, Morris water maze laboratory escape latency and average detection range are more normally organized and have been increased by 273.21% and 153.76% respectively, stopping in the diving tower experiment obviously increases by 502.85% and 497.63% than normal group minimizing 47.87%, escape latency and errors number incubation period, obstacle appears in the memory consolidation, and the study and work memory ability significantly descends.The mice Morris water maze laboratory escape latency of respectively organizing that gives baicalin and taurine compositions reduces than model group in various degree with average detection range; the diving tower experiment stops and improves than model group incubation period; escape latency and errors number reduce, and action effect has the trend that is better than the positive control drug aricept.The experimental result prompting; baicalin can promote the interior macromole relevant with longterm memory of brain with the compositions that the taurine proportioning forms, and particularly novel protein is synthetic; improve memory and consolidate learning and memory and the cognitive function of obstacle mice, can excellent effects ground be used for prevention and improve the learning and memory infringement or improve dementia symptom, control dementia.
The D-galactose forms excessive ultra-oxygen anion free radical in metabolic process, can produce the peroxidating infringement to tissue, makes body aging, causes that a series of degeneration of cerebral tissue and neuron change, and this process and senile dementia are in close relations.Therefore by the dementia due to the energy simulating nature aging of animal injection of d-galactose or free-radical oxidation damage etc.Through the test data contrast as can be known, after the mouse subcutaneous injection D-galactose, Morris water maze laboratory escape latency and average detection range are more normally organized and have been increased by 383.92% and 625.41% respectively, stopping in the diving tower experiment obviously increases by 221.57% and 921.66% than normal group minimizing 57.72%, escape latency and errors number incubation period, illustrates that ability of learning and memory in mice significantly descends.The compositions that forms with 6: 1 proportionings, 18: 1 proportionings, 2: 1 proportionings with baicalin and taurine is irritated after stomach gives mice; mice Morris water maze laboratory escape latency and average detection range all reduce in various degree than model group, and diving tower escape latency and errors number also obviously reduce and stop and improve in various degree incubation period.In the Morris water maze laboratory; compositions and 3 groups of effects of compositions that baicalin and taurine 6: 1 and 2: 1 proportionings form are suitable with the positive drug aricept; the compositions that 18: 1 proportionings form makes mice escape latency and average detection range reduce 68.42% and 69.18% respectively than model group, action effect even be better than the positive drug aricept.In the diving tower experiment, the compositions that 6: 1 proportioning high doses of baicalin and taurine, middle dosage and 18: 1 proportionings form make mice stop incubation period than model components you can well imagine high by 93.87%, 67.52% and 81.45%, escape latency reduces 61.88%, 44.79 and 55.32% respectively, and the trend that is better than positive drug aricept (stop to improve 45% incubation period, escape latency reduce 30.14%) is arranged; Its diving tower errors number reduces 52.83%, 27.46 and 25.68% respectively.Experimental result shows; the compositions that baicalin and taurine form with certain proportioning can enhancing body oxidation resistance; inhibited oxidation radical pair body brain tissue impairment; resist old and feeble and the protection brain cell; improve learning and memory and cognitive function that deterioration of neurons and radical damage cause the brain function decline mice, point out it to can be used for improving dementia symptom, control dementia.
In sum; whole body zoopery by cell in vitro experiment and four kinds of different pathological mechanism models proves that baicalin of the present invention and taurine composition can the neuroprotective cell injury, can improve the ability of learning and memory that synthesizes dementia due to minimizing, aging and the radical damage because of new albumen in A β deposition, cholinergic nerve function damage, the brain.And effect is better than baicalin or taurine uses separately.
Although the dull-witted cause of disease and pathomechanism complexity are multiple, or there is a multiple theory, for example cerebrovascular shows effect repeatedly, " A β theory ", " neuron loss theory ", " neurofibrillary tangles theory (as Tua abnormal protein phosphorylation) ", " the unusual theory of neurotransmitter (reducing as acetylcholine Ach) ", " toxicity of excitatory amino acid theory (as glutamic acid Glu hyperfunctioning) ", " free radical and oxidative damage theory ", " abnormal carbohydrate metabolism theory " etc., but the final pathology damage of the various dementias due to them and clinical symptoms performance are all roughly the same, final all is by one group of syndrome chronic or the senior cerebral disorder that carrying out property brain damage causes, all with memory, thinking, identification, language, one group of obviously going down to characteristics of senior cerebral nerve cognitive functions such as judgement is chronic, carry out the neural mental deterioration disease of sexual development.Therefore; by the experimental result of lifting among the present invention, those skilled in the art can reasonably expect: the cause of disease outside baicalin of the present invention and taurine composition are listed to the present invention and the ability of learning and memory of the various senile dementias due to the pathomechanism and cognitive function all have the improvement effect.
In sum, baicalin of the present invention and taurine composition can be used for improving learning and memory and cognitive dysfunction, and senile dementia is had the better prevention effect.The cause of disease according to different dementia is different with pathogenesis; baicalin of the present invention and taurine composition mainly refer to refer to Alzheimer dementia (AD) especially for control Alzheimer dementia (AD), vascular dementia (VaD), Combination dementia (MixD).The different dementia hypotype that forms because of the different causes of disease and pathogenesis has roughly the same final pathology damage and clinical symptoms performance (as memory impairment etc.) again; therefore expect that baicalin of the present invention and taurine composition also can be used for improving memory ability and the cognitive function of the dementia due to other various causes of disease and the different pathological mechanism, improve the symptom of following any or several dementias: Louis corpusculum dementia (DLB); volume temporal lobe dementia (FTD); dementia in Parkinson disease (PD); dementia in Creutzfeldt-Jakob disease (CJD); normal pressure hydrocephalus dementia (NPH); Wei-Ke two formula syndrome dementias (WKS).
And medical science and study of pharmacy personnel can't not do related experiment and not analyze under the prerequisite in conjunction with several pathomechanism model generalizations in advance, learn that in advance baicalin and taurine composition have effect and the said medicine purposes thereof of improving learning and memory and cognitive dysfunction and control senile dementia.
The specific embodiment
Described baicalin, taurine all can be bought for market, also can adopt existing extraction, separation, purification or technical method preparation such as synthetic and get.As: get Radix Scutellariae, decoct with water secondary, merging filtrate concentrates, and the hydrochloric acid adjust pH is 1~2, and under about 75 ℃ of conditions, the insulation precipitation is separated out baicalin; The collecting precipitation thing adds aqueous dispersion, and NaOH solution adjust pH is 7, adds the equal-volume dissolve with ethanol, filters, and filtrate is 1~2,70 ℃ of insulation 1h with the hydrochloric acid adjust pH, leaves standstill secondary precipitation; Precipitate gets the baicalin crude product with ethanol cyclic washing, drying; Crude product adds aqueous dispersion, heat temperature raising to 75 ℃, NaOH solution adjust pH is about 7, makes its dissolving, the hydrochloric acid adjust pH is 6, add ethanol and make and contain the alcohol amount and reach about 70%, filtrate is 2 with the hydrochloric acid adjust pH, staticly settles, get the baicalin elaboration, use methanol reflux repeated crystallization again, get the brilliant baicalin of faint yellow fine needle, purity 〉=98%.
Baicalin in following examples, taurine are market and buy, and purity all 〉=98%.
Embodiment 1
Baicalin (115.2g), taurine (76.8g) are mixed with 3: 2 ratio, get baicalin and taurine composition (192g) behind stirring, grinding, the mix homogeneously.
Embodiment 2
Baicalin (144g), taurine (24g) are mixed with 6: 1 ratio, get baicalin and taurine composition (168g) behind stirring, grinding, the mix homogeneously.
Embodiment 3
Baicalin (180g), taurine (20g) are mixed with 9: 1 ratio, get baicalin and taurine composition (200g) behind stirring, grinding, the mix homogeneously.
Embodiment 4
Baicalin (180g), taurine (10g) are mixed with 18: 1 ratio, get baicalin and taurine composition (190g) behind stirring, grinding, the mix homogeneously.
Embodiment 5
Baicalin (108g), taurine (54g) are mixed with 2: 1 ratio, get baicalin and taurine composition (162g) behind stirring, grinding, the mix homogeneously.
Embodiment 6
Baicalin (42.8g), taurine (2g) are mixed with 21.4: 1 ratio, get baicalin and taurine composition (44.8g) behind stirring, grinding, the mix homogeneously.
Provide some preferred embodiments of baicalin of the present invention, taurine percentage by weight below in conjunction with table 1; but the content of both compositions of the present invention is not limited to listed numerical value in this table; for a person skilled in the art, can in table, rationally summarize and reasoning on the basis of listed numerical range fully.
Table 1 baicalin, taurine account for the percentage by weight (%) of compositions
Figure BSA00000869346700081
Experimental data 1: baicalin and taurine composition are to A β 25-35Cause the protective effect of SH-SY5Y cell injury
(1) determine that baicalin, taurine are to the non-toxic concn scope of SH-SY5Y cell:
Human neuroblastoma cell SH-SY5Y is with every hole 1 * 10 4Individual cell is seeded in 96 orifice plates, and in 37 ℃, volume fraction is 5% C0 with the DMEM/F12 culture fluid that contains 10% hyclone 2Cultivate in the incubator.Treat that cell confluency reaches at 80% o'clock, change culture fluid into contain baicalin or taurine (5,10,20,40,80,160 μ mol/L) serum-free medium and continue to cultivate 24 hours.Adopt mtt assay and LDH method to measure cytoactive.The result shows that baicalin, taurine are in 0~160 μ mol/L scope, to the equal avirulence of SH-SY5Y cell.
(2) determine that baicalin, taurine are separately to A β 25-35Cause the optimum concentration of SH-SY5Y cell injury performance protective effect:
The SH-SY5Y cell is with every hole 1 * 10 4Individual cell is seeded in 96 orifice plates, and in 37 ℃, volume fraction is 5% C0 with the DMEM/F12 culture fluid that contains 10% hyclone 2Cultivate in the incubator.Treat that cell confluency reaches at 80% o'clock, culture fluid is changed into contain baicalin or taurine (5,10,20,40,80 μ mol/L) and 20 μ mol/L condensed state A β respectively 25-35Serum-free medium continue to cultivate 24 hours.Mtt assay is measured cytoactive.Calculate the relative survival rate of respectively organizing cell.The results are shown in Table 2, table 3.
(3) baicalin and taurine composition cause the protective effect of SH-SY5Y cell injury to A β 25-35:
Baicalin and the taurine composition of getting embodiment 2, embodiment 4, embodiment 5, embodiment 6 acquisitions are used for experiment, abbreviate " compositions 1 ", " compositions 2 ", " compositions 3 ", " compositions 4 " respectively as.The SH-SY5Y cell is with every hole 1 * 10 4Individual cell is seeded in 96 orifice plates, and in 37 ℃, volume fraction is to cultivate in 5% the CO2 incubator with the DMEM/F12 culture fluid that contains 10% hyclone.Cell divides 8 groups: normal control group, model control group, " compositions 1 " group (wherein contain baicalin 24 μ mol/L; taurine 14 μ mol/L), " compositions 2 " group (wherein contains baicalin 40 μ mol/L; taurine 8 μ mol/L), " compositions 3 " group (wherein contains baicalin 9 μ mol/L; taurine 16 μ mol/L), " compositions 4 " group (wherein containing baicalin 48 μ mol/L, taurine 8 μ mol/L), baicalin matched group (80 μ mol/L), taurine group (20 μ mol/L).Treat respectively to organize cell confluency and reach at 80% o'clock, culture fluid is changed into contain relative medicine and 20 μ mol/L condensed state A β 25-35Serum-free medium continue to cultivate 24 hours.Mtt assay is measured cytoactive.Calculate and respectively organize the cell relative survival rate.The results are shown in Table 4.
Table 2 baicalin is to A β 25-35Cause the protective effect (n=5) of SH-SY5Y cell injury
Figure BSA00000869346700101
Compare * P<0.05, * * P<0.01 with model group.
Table 3 taurine is to A β 25-35Cause the protective effect (n=5) of SH-SY5Y cell injury
Figure BSA00000869346700102
Compare * P<0.05, * * P<0.01 with model group.
Table 4 baicalin and taurine composition are to A β 25-35Cause the protective effect (n=5) of SH-SY5Y cell injury
Figure BSA00000869346700103
Figure BSA00000869346700111
Compare * P<0.05, * * P<0.01 with model group.
By table 2, table 3 result as can be known, baicalin, taurine all have significant protective effect to the SH-SY5Y cell injury in the test dose scope.Compare with the model control group cell survival rate, baicalin, taurine are separately to A β 25-35The optimum concentration that causes SH-SY5Y cell injury performance protective effect is respectively 40~80 μ mol/L, 10~80 μ mol/L.
By table 4 result as can be known, baicalin all significantly improves than model group with each compositions group SH-SY5Y cell relative survival rate that taurine is formed with different proportionings.Wherein " compositions 2 ", " compositions 4 " group cell survival rate have improved (90.24 ± 7.46) %, (114.39 ± 10.79) % respectively than model control group; be higher than baicalin matched group (68.62 ± 11.54) % and taurine matched group (83.92 ± 20.17) %; illustrate that compositions has stronger protective effect to neural cell injury, and the protective effect when using separately than baicalin or taurine is more remarkable.
Experimental data 2: baicalin and taurine composition are to A β 1-42Intracerebroventricular causes the influence of dementia rats model learning memory
Baicalin and the taurine composition of getting embodiment 2, embodiment 4, the acquisition of embodiment 5 preparation methoies are used for experiment, abbreviate " compositions 1 ", " compositions 2 ", " compositions 3 " respectively as.
Establish 10 experimental grouies altogether: the normal control group; model group; positive drug group (aricept 0.9mg/kg); " compositions 1 " high, medium and low dosage group (280,140,70mg/kg); " compositions 2 " group (190mg/kg); " compositions 3 " group (90mg/kg), baicalin matched group (240mg/kg), taurine matched group (40mg/kg).
The SD rat, body weight (250 ± 10) g, intracerebroventricular A β 1-42Modeling, the normal saline of normal control group injection equivalent.Carry out the experiment of Morris water maze and diving tower after one week of modeling and carry out the detection of learning and memory behavioristics, whether successful to estimate modeling.After the model evaluation success, the animal random packet, the medicine group is irritated stomach and is given corresponding medicine, and normal control group and model group give the normal saline of equivalent, successive administration 30 days.The learning and memory in rats ability is respectively organized in Morris water maze and the measuring of rat diving tower.The results are shown in Table 5, table 6.
Table 5 compositions is to A β 1-42Intracerebroventricular causes the influence (n=10) of dementia rats Morris water maze laboratory
Figure BSA00000869346700121
Figure BSA00000869346700131
Compare * P<0.05, * * P<0.01 with model group.
Table 6 compositions is to A β 1-42Intracerebroventricular causes the influence (n=10) of dementia rats diving tower experiment
Figure BSA00000869346700132
Compare * P<0.05, * * P<0.01, Δ P<0.001 with model group.
Model group rat Morris water maze laboratory escape latency, average search distance significantly increase, and escape latency and errors number also significantly increase in the diving tower experiment, and rat intracerebroventricular A β is described 1-42After show tangible dementia symptom, the remarkable obstacle of cognitive learning ability.By Morris water maze laboratory result in the table 5 as can be known: trained the 3rd day, positive drug group, " compositions 1 " high dose group, " compositions 2 " group rat escape latency obviously are less than model group; To the 4th day of training, the escape latency that " compositions 1 " high, medium and low dosage group, " compositions 2 " reach " compositions 3 " group rat all obviously was less than model group, and baicalin matched group and taurine matched group and model group are more as broad as long and arrive at this moment; To training the 5th day; positive drug group and compositions 1 height, middle dosage, 2 groups of rat average search distances of compositions also reduce in various degree than model group, and arrive this moment the baicalin matched group and taurine control rats average search apart from relatively there is no significant difference with model group.Training the 7th day, each medicine group all reduces in various degree than model group rat escape latency, average search distance each medicine group except the taurine matched group also reduces in various degree than model group, and is wherein remarkable with 2 groups of effects of compositions, suitable with positive drug aricept effect; And compositions 1 high dose group, compositions obviously are better than baicalin matched group and taurine matched group for 2 groups.Compositions 1 high dose group, 2 groups of compositionss, baicalin matched group and taurine matched group and model group are relatively; Morris water maze laboratory escape latency reduces 44.83,62.69%, 30.46% and 15.45% respectively; the average search distance reduces 35.47%, 48.17%, 16.83% and 5.3% respectively.In the compositions 1, though 3 groups of low dose group and compositionss do not have marked difference than baicalin matched group and taurine matched group effect, and the trend of excellent effect is also arranged.There is a certain amount of effect relationship in influence to rat water maze ability of learning and memory between " compositions 1 " height, the middle dosage group; In, low dose group escapes the influence hide to rat and do not show dose-effect relationship, relies on trend but the influence of rat average search distance is presented certain dose-effect.In the diving tower experiment: each medicine group rat stops obviously to be increased than model group incubation period, and escape latency significantly reduces, and each compositions group is better than baicalin contrast and taurine matched group; Each medicine group diving tower errors number also reduces in various degree than model group, and 2 groups of compositions 1 high dose group and compositionss also reduce in various degree than baicalin contrast and taurine matched group errors number; The overall merit diving tower is tested each index, and is the most remarkable with compositions 1 high dose group and compositions 2 effects, suitable with positive drug aricept action effect, even compositions has the trend that is better than aricept for 2 groups.Compositions 1 high dose group, 2 groups of compositionss, baicalin matched group and taurine matched group and model group are relatively; diving tower stops increases by 249.25%, 270.93%, 154.75% and 142.37% respectively incubation period; escape latency has reduced 91.94%, 99.54%, 56.79% and 51.55% respectively, and errors number reduces 52.85%, 72.31%, 28.75% and 41.91% respectively.Rat cognitive function and learning and memory test experiments result show that the baicalin in the experiment and taurine composition can reduce A β 1-42Cause dementia rats water maze escape latency and average search distance, increase the experiment of rat diving tower stop incubation period, reduce escape latency and the errors number of diving tower experiment, and compositions is rapid-action and effect is more obvious; Baicalin and taurine composition that explanation is formed with certain proportioning can improve cognitive function and the ability of learning and memory that A β causes dementia rats, and action effect is better than baicalin and taurine acts on separately.Point out it can be effectively to be used for improving learning and memory and cognitive dysfunction, control dementia.
Experimental data 3: baicalin and taurine composition cause the memory improvement effect of memory acquisition disturbance mouse model to scopolamine
Baicalin and the taurine composition of getting embodiment 2, embodiment 4, the acquisition of embodiment 5 preparation methoies are used for experiment, abbreviate " compositions 1 ", " compositions 2 ", " compositions 3 " respectively as.
Establish 8 experimental grouies altogether: normal control group, model group, positive drug group (aricept 1.6mg/kg), " compositions 1 " high, medium and low dosage group (560,280,140mg/kg), " compositions 2 " group (380mg/kg), " compositions 3 " group (180mg/kg).
The KM mice, body weight (25 ± 2) g presses the modeling of 1mg/kg dosage lumbar injection scopolamine, the normal saline of normal control group injection equivalent.Carry out the experiment of Morris water maze and diving tower after one week and carry out the detection of learning and memory behavioristics, whether successful to estimate modeling.After the model evaluation success, the animal random packet, the medicine group is irritated stomach and is given corresponding medicine, and normal control group and model group give the normal saline of equivalent, successive administration 30 days.Ability of learning and memory in mice is respectively organized in Morris water maze and the measuring of mice diving tower.The results are shown in Table 7, table 8.
Table 7 compositions causes the influence (n=10) of memory acquisition disturbance mice Morris water maze laboratory to scopolamine
Figure BSA00000869346700151
Compare * P<0.05, * * P<0.01 with model group.
Table 8 compositions causes the influence (n=10) of memory acquisition disturbance mice diving tower experiment to scopolamine
Figure BSA00000869346700161
Compare * P<0.05, * * P<0.01, Δ P<0.001 with model group.
Model group KM mice Morris water maze laboratory escape latency and average detection range significantly increase than normal group, stopping obviously reduction incubation period, escape latency and errors number in the diving tower experiment also significantly increases, after KM mouse peritoneal injection scopolamine is described, blocked cholinergic nerve path m receptor, suppress its cholinergic nerve function, make mice the study and work memory impairment occur.After giving baicalin and taurine composition, KM mice Morris water maze laboratory escape latency and average detection range reduce in various degree than model group, wherein compositions 1 height, middle dosage group, compositions 2,3 groups of mices of compositions reduce 52.76%, 39.54,73.53% and 26.62% in the 7th day escape latency of training respectively than model group, and the average search distance reduces 39.91%, 30.75%, 47.98% and 24.61% respectively; In the diving tower experiment, compositions 1 high dose group, compositions 2,3 groups of mices of compositions stop and increase by 197.66%, 208.43%, 85.67% respectively than model group incubation period, escape latency reduces 65.72%, 71.03%, 28.65% respectively, and errors number reduces 62.55%, 57.40%, 30.52% respectively.Experimental result shows; the compositions that baicalin and taurine proportioning form can be improved brain cholinergic nerve of centrum function; improvement causes study and work memory ability and the cognitive competence of memory acquisition disturbance mice because cholinergic nerve is impaired, the prompting baicalin can be used for improving the relevant memory impairment of cholinergic and control senile dementia with taurine composition.
Experimental data 4: baicalin and taurine composition cause the memory improvement effect that the obstacle mouse model is consolidated in memory to cycloheximide
Baicalin and the taurine composition of getting embodiment 2, embodiment 4, the acquisition of embodiment 5 preparation methoies are used for experiment, abbreviate " compositions 1 ", " compositions 2 ", " compositions 3 " respectively as.
Establish 8 experimental grouies altogether: normal control group, model group, positive drug group (aricept 1.6mg/kg), " compositions 1 " high, medium and low dosage group (560,280,140mg/kg), " compositions 2 " group (380mg/kg), " compositions 3 " group (180mg/kg).
The KM mice, body weight (25 ± 2) g, the modeling of lumbar injection cycloheximide, the normal saline of normal control group injection equivalent.Carry out the experiment of Morris water maze and diving tower after one week and carry out the detection of learning and memory behavioristics, whether successful to estimate modeling.After the model evaluation success, the animal random packet, the medicine group is irritated stomach and is given corresponding medicine, and normal control group and model group give the normal saline of equivalent, successive administration 30 days.The improvement situation of ability of learning and memory in mice is respectively organized in Morris water maze and the measuring of mice diving tower.The results are shown in Table 9, table 10.
Table 9 compositions causes the influence (n=10) that obstacle mice Morris water maze laboratory is consolidated in memory to cycloheximide
Figure BSA00000869346700171
Figure BSA00000869346700181
Compare * P<0.05, * * P<0.01 with model group.
Table 10 compositions causes the influence (n=10) that the experiment of obstacle mice diving tower is consolidated in memory to cycloheximide
Figure BSA00000869346700182
Compare * P<0.05, * * P<0.01, Δ P<0.001 with model group.
New albumen synthesizes have middle important function in learning and memory, macromole protein and RNA synthetic closely related particularly in consolidation of memory and reproduction and the brain.The inferior amide of hexamethylene is a kind of protein synthesis inhibitor, can cause the synthetic minimizing of albumen relevant with longterm memory in the brain, has hindered the formation of memory, and has caused study and the cognitive dysfunction of brain.Model group mice Morris water maze laboratory escape latency and average detection range are more normally organized and have been increased by 273.21% and 153.76% respectively, stopping in the diving tower experiment obviously increases by 502.85% and 497.63% than normal group minimizing 47.87%, escape latency and errors number incubation period, after the inferior amide of mouse peritoneal injection hexamethylene was described, ability of learning and memory significantly descended.The mice Morris water maze laboratory escape latency of respectively organizing that gives baicalin and taurine compositions reduces than model group in various degree with average detection range; wherein 3 groups of effects of dosage group and compositions are similar to the positive drug aricept in the compositions 1; the most remarkable with compositions 1 high dose group and 2 groups of effects of compositions, be better than other each pharmaceutical intervention group.The mice diving tower experiment respectively organized that gives baicalin and taurine composition stops and improves than model group incubation period, and escape latency and errors number reduce, wherein compositions 1 effect of high dosage effect even the trend that is better than the positive drug aricept is arranged.This experimental result prompting; baicalin can promote the interior macromole relevant with longterm memory of brain with the compositions that the taurine proportioning forms, and particularly novel protein is synthetic; improve memory and consolidate learning and memory and the cognitive function of obstacle mice, can excellent effects ground be used for prevention and improve the learning and memory infringement or improve dementia symptom, control dementia.
Experimental data 5: baicalin and taurine composition cause the memory improvement effect of brain aging mouse model to the D-galactose
Baicalin and the taurine composition of getting embodiment 2, embodiment 4, the acquisition of embodiment 5 preparation methoies are used for experiment, abbreviate " compositions 1 ", " compositions 2 ", " compositions 3 " respectively as.
Establish 8 experimental grouies altogether: normal control group, model group, positive drug group (aricept 1.6mg/kg), " compositions 1 " high, medium and low dosage group (560,280,140mg/kg), " compositions 2 " group (380mg/kg), " compositions 3 " group (180mg/kg).
The KM mice, body weight (25 ± 2) g, the modeling of subcutaneous injection D-galactose, the normal saline of normal control group injection equivalent.Carry out the experiment of Morris water maze and diving tower after one week and carry out the detection of learning and memory behavioristics, whether successful to estimate modeling.After the model evaluation success, the animal random packet, the medicine group is irritated stomach and is given corresponding medicine, and normal control group and model group give the normal saline of equivalent, successive administration 30 days.The improvement situation of ability of learning and memory in mice was respectively organized in Morris labyrinth and the measuring of mice diving tower after administration finished.The results are shown in Table 11, table 12.
Table 11 compositions causes the influence (n=10) of brain aging mice Morris water maze laboratory to the D-galactose
Figure BSA00000869346700191
Figure BSA00000869346700201
Compare * P<0.05, * * P<0.01 with model group.
Table 12 compositions causes the influence (n=10) of brain aging mice diving tower experiment to the D-galactose
Figure BSA00000869346700202
Compare * P<0.05, * * P<0.01, Δ P<0.001 with model group.
The D-galactose forms excessive ultra-oxygen anion free radical in metabolic process, tissue is produced the peroxidating infringement, makes body aging, causes that a series of degeneration of cerebral tissue and neuron change, and this process and senile dementia are in close relations.Find out from experiment, model group mice Morris water maze laboratory escape latency and average detection range are more normally organized and have been increased by 383.92% and 625.41% respectively, stopping in the diving tower experiment obviously increases by 221.57% and 921.66% than normal group minimizing 57.72%, escape latency and errors number incubation period, after mouse subcutaneous injection D-galactose was described, ability of learning and memory significantly descended.The mice Morris water maze laboratory escape latency of respectively organizing that gives baicalin and taurine composition all reduces than model group in various degree with average detection range, wherein 2 groups of minimizings of compositions degree is the most remarkable, its escape latency and average detection range reduce 68.42% and 69.18% respectively than model group, action effect even be better than the positive drug aricept; It is suitable with the positive drug aricept that compositions 1 height, middle dosage group and 3 groups of escape latency of compositions reduce degree.In the diving tower experiment, 2 groups of compositions 1 height, middle dosage group and compositionss stop incubation period than model components you can well imagine high by 93.87%, 67.52% and 81.45%, escape latency reduces 61.88%, 44.79 and 55.32% respectively, and the trend that is better than positive drug aricept (stop to improve 45% incubation period, escape latency reduce 30.14%) is arranged; Its diving tower errors number reduces 52.83%, 27.46 and 25.68% respectively.Experimental result shows; the oxidation resistance of the compositions energy enhancing body that baicalin and taurine proportioning form; the damage of inhibited oxidation radical pair body tissue; resist old and feeble and the protection brain cell; improve learning and memory and the cognitive function of deterioration of neurons and brain function decline mice, point out it to can be used for improving dementia symptom, control dementia.

Claims (10)

1. a baicalin and taurine composition are for the protection of the medicinal usage of neural cell injury.
2. a baicalin and taurine composition are used for improving the medicinal usage of ability of learning and memory.
3. a baicalin and taurine composition are used for improving the medicinal usage of cognitive dysfunction.
4. according to each purposes of claim 1~3, it is characterized in that described compositions for the preparation of prevention and treat the purposes of dull-witted medicine.
5. according to the purposes of claim 4, it is characterized in that described compositions is for the preparation of the purposes of preventing and treating Alzheimer dementia (AD), vascular dementia (VaD), Combination dementia (MIXD), Louis corpusculum dementia (DLB), volume temporal lobe dementia (FTD), dementia in Parkinson disease (PD), dementia in Creutzfeldt-Jakob disease (CJD), normal pressure hydrocephalus dementia (NPH), Wei-Ke two formula syndrome dementia (WKS) medicines.
6. according to the purposes of claim 5, it is characterized in that described compositions is for the preparation of the purposes of preventing and treating Alzheimer dementia (AD), vascular dementia (VaD), Combination dementia (MixD) medicine.
7. according to the purposes of claim 6, it is characterized in that described compositions is for the preparation of the purposes of prevention and treatment Alzheimer dementia (AD) medicine.
8. according to each described purposes of claim 1~7, wherein said baicalin and taurine composition are made up of the following weight proportioning: baicalin 99.9%~60.0%, taurine 0.1~40.0%.
9. purposes according to claim 8, wherein said baicalin and taurine composition are made up of the following weight proportioning: baicalin 99.0%~90.0%, taurine 1.0%~10.0%.
10. pharmaceutical preparation; the main effective ingredient that it is characterized in that described pharmaceutical preparation is baicalin and taurine composition; and with one or more pharmaceutically acceptable excipient and/or can be prepared from the other medicines of its compatibility; wherein said baicalin and taurine composition are made up of the following weight proportioning: baicalin 99.9%~60.0%, taurine 0.1~40.0%.
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