CN101869600A - Medicinal composition for treating degenerative change in central nervous system, preparation method and application thereof - Google Patents
Medicinal composition for treating degenerative change in central nervous system, preparation method and application thereof Download PDFInfo
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- CN101869600A CN101869600A CN 201010217290 CN201010217290A CN101869600A CN 101869600 A CN101869600 A CN 101869600A CN 201010217290 CN201010217290 CN 201010217290 CN 201010217290 A CN201010217290 A CN 201010217290A CN 101869600 A CN101869600 A CN 101869600A
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- rhizoma chuanxiong
- angelicae sinensis
- radix angelicae
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Abstract
The invention belongs to the technical field of pharmacy, in particular provides to a medicinal composition, which mainly comprises the following raw material medicaments: baical skullcap root, szechuan lovage rhizome and Chinese angelica. The invention also provides a preparation method and application of the medicinal composition. Through the verification of experiments, the medicinal composition has the effects of detoxifying and removing obstruction in channels and promoting mentality and inducing resuscitation, can improve the cognitive function of dementia or cognitive disorder effectively, improve behavioral anomaly and the life quality of patients and aim at oxidative stress damage in brain cells and micro-inflammatory damage effectively, and has the effect of treating the degenerative change in the central nervous system, particularly senile dementia or vascular dementia or mild cognitive disorder.
Description
Technical field
The present invention relates to a kind of pharmaceutical composition, this pharmaceutical composition is mainly by Radix Scutellariae, Rhizoma Chuanxiong with when being grouped into, effect with treatment and alleviation central nervous system degeneration, especially treat and alleviate the effect of senile dementia, vascular dementia or mild cognitive impairment disease, belong to technical field of Chinese medicines.
Background technology
Alzheimer disease is a kind of common central nervous system degenerative disease.In recent years, along with the aging of world population. the sickness rate of senile dementia is continuous ascendant trend, at present should the 4th of the disease row cause of death (being only second to heart disease, tumor and apoplexy), become one of the severeest problem that current geriatrics faces.According to statistics, China more than 60 years old in the population prevalence about 5%, about 6,000,000, and increase and increase with the age.The pathological change of alzheimer disease mainly shows as the obvious atrophy of cerebral cortex, based on frontal lobe, temporal lobe, top on form.Convolutional atrophy narrows down, the broadening of brain ditch, and the ventricles of the brain enlarge; It is to have occurred assembling the amyloid plaques (senile plaque) that forms (SP) by the hypotype (being A β 42) of 42 amino acid peptides of amyloid-beta outside brain cell that main histopathology changes, and has occurred the neurofibrillary tangles (NFT) that the Tau albumen by cell microtubule and excessive phosphorylation constitutes in neuron.Because the generation of amyloid-beta increases or removes minimizing, accumulative amyloid-beta has started a series of pathological processes in nervus retrogression degeneration and dementia are fallen ill, i.e. amyloid hypothesis.The gathering of A β 42 has caused the formation of amyloid plaques, and start a series of and neuron and synapse dysfunction, inflammatory reaction, the proteic excessive phosphorylation of Tau, neurofibrillary tangles formation, neuronal death, the unusual relevant change of neurotransmitter, finally cause dull-witted generation.Alzheimer disease is clinical to serve as main performance with confusion, memory defects, personality and aphasis.This disease can cause the life of elderly person downgrade, severe patient forfeiture self care ability brings heavy burden for family, society, along with the aging of China's population, its sickness rate also continues to raise, and therefore actively preventing and treating alzheimer disease has become an old clinical medical important topic.
By pathogenesis, senile dementia can be divided into 4 classes: and Alzheimer's disease (Alzheimer ' S diease, AD), vascular dementia (Vascular dementia, VD), mixed type dementia (Mixed dementia, MD) and other dementia (Other dementia, OD), wherein AD and VD are the most common.
Modern medicine has carried out years of researches to the cause of disease and the pathogenesis of alzheimer disease, but because of its cause of disease complexity, so far its concrete link is not understood as yet, the cause of disease and pathogenetic understanding to AD mainly concentrates on chronic inflammatory disease and the decline of carrying out property of cholinergic system function in the h and E caused by factors brain at present, and main more representative theory has: the hemorheology theory of cholinergic theory, radical damage theory, calcium overload theory and the traditional Chinese medical science etc.
Chinese medicine has original understanding to senile dementia disease position, the cause of disease, pathogenesis.The differentiation of tcm theory thinks, no matter vascular dementia or Alzheimer all mainly show as the traditional Chinese medical science " god's " pathological changes.The traditional Chinese medical science thinks that expectorant is turbid, blood stasis is to quicken the key factor that brain aging causes senile dementia, and it is the core of morbidity that heart kidney deficiency, Qi and blood deficiency, the moon decrease smart declining, and the key factor that the effect of expectorant stasis of blood pathogenic fire is just fallen ill.Determination of treatment based on pathogenesis obtained through differentiation of symptoms and signs is the characteristic and the elite of the traditional Chinese medical science, occupies an leading position in the treatment of the traditional Chinese medical science to alzheimer disease.
Because of there is multiple hypothesis in the pathogenesis of alzheimer disease, still there is not at present the explanation alzheimer disease that a kind of hypothesis can be more complete, although therefore the medicament categories of clinical treatment alzheimer disease is a lot, but also do not have specific medicament, the medicine that is used for the treatment of alzheimer disease at present mainly contains and improves cholinergic nerve and transmit medicine, free radical scavenger and antioxidant, improve blood circulation and metabolic medicine of brain cell and calcium channel blocker etc.
In the treatment of alzheimer disease, Chinese medicine and compound recipe thereof have abundant theory and practice experience aspect the preventing and treating of aging and diseases associated with senescence alleviating, advantage with whole conditioning, comprehensive measure, and toxic and side effects is few, suit to take for a long time, especially the many target spots of Chinese medicine, too many levels, multimode mechanism of action are treated AD and are had obvious superiority, in the AD treatment, play more and more important effect.
Chinese medicine compound is the mechanism of action according to Chinese medical theory, selects different prescriptions for use, adopts multiple analysis means, from multi-faceted, multi-angle, has set forth the nootropic effect of compound Chinese medicinal preparation comprehensively and objectively.According to clinical experience and experimentation, effective Chinese medicine compound that option goes out has: the kidney invigorating Fructus Alpiniae Oxyphyllae side is that main biochemical indicator, the Chinese medicine the kidney invigorating Fructus Alpiniae Oxyphyllae side of alzheimer disease has the kidney invigorating, QI invigorating, gives birth to marrow, supports the brain effect, and its pastille serum can be protected these changes that caused by amyloid effectively; Danzhi Xiaoyao San reduces rat model brain-capacity load, protein, alditol reductase courier content; Brain protein, the alditol reductase courier content of model are gone up, alleviate the substance metabolism obstacle.The Chinese medicine alzheimer disease has accumulated rich experience, advantage with whole conditioning, comprehensive measure, and toxic and side effects is few, suits to take for a long time, at aspects such as health care prevention and harmonizing emotions many distinctive features are arranged simultaneously, formed the comparatively theory of system.But also come with some shortcomings at present, mainly show and do not adopt unified diagnosis, typing, efficacy assessment standard in many clinical researches, at prescription, select on medicine, the dosage and vary with each individual, can't standard.The treatment alzheimer disease should be puted prevention first with early diagnosis, and develops the medicine series of determined curative effect early, impels the research of Chinese medicine alzheimer disease deeply to change with scientific, improves the clinical efficacy of Chinese medicine alzheimer disease, to promote the well-being of mankind.Along with the change Chinese medicine of the aging of population and spectrum of disease can be more and more wide in the prospect of preventing and treating alzheimer disease.
Still do not have bibliographical information at present and only be prepared into preparation, especially these three kinds of medicines are share and be used for the treatment of central nervous system's degeneration, do not see correlational study by Radix Scutellariae, Rhizoma Chuanxiong, Radix Angelicae Sinensis three flavor crude drug.
Summary of the invention
The object of the present invention is to provide a kind of pharmaceutical composition, this pharmaceutical composition is mainly by Radix Scutellariae, Rhizoma Chuanxiong with when being grouped into, have treatment and alleviate the effect of nervus centralis degeneration, especially have treatment and alleviate the effect of senile dementia, vascular dementia and mild cognitive impairment.
Preferably, in the pharmaceutical composition of the present invention, the weight proportion of described primary raw material medicine Radix Scutellariae, Rhizoma Chuanxiong and Radix Angelicae Sinensis is Radix Scutellariae 5-20 part, Rhizoma Chuanxiong 5-15 part, Radix Angelicae Sinensis 5-15 part; More preferably, the weight proportion of described primary raw material medicine is Radix Scutellariae 10-15 part, Rhizoma Chuanxiong 8-12 part, Radix Angelicae Sinensis 8-12 part, Radix Scutellariae 12-15 part more preferably, Rhizoma Chuanxiong 10-12 part, Radix Angelicae Sinensis 10-12 part.
The present invention also aims to provide a kind of preparation method for preparing active ingredient in pharmaceutical of the present invention, when preparing active ingredient in pharmaceutical of the present invention, Rhizoma Chuanxiong and Radix Angelicae Sinensis volatile ingredient are obtained in preparation from crude drug earlier, and be standby; Rhizoma Chuanxiong, Radix Angelicae Sinensis and Radix Scutellariae clear paste mix homogeneously behind gained volatile ingredient and the organic solvent extraction that dissolves each other through water or with water, promptly get the pharmaceutical composition active component, mixed active component adds acceptable auxiliary in the pharmacy, prepare required drug unit dosage form with the preparation routine techniques, as being clinical required dosage forms such as granule, tablet, capsule, drop pill, oral liquid, suspension, emulsion, injection.
Perhaps, the extraction of active ingredients preparation method of primary raw material drug composition of the present invention can be: the crude drug of getting recipe quantity, after pulverizing merging, the organic solvent extraction that dissolves each other with water or with water 1-4 time, filter, merge each time and extract filtrate, the active component extract clear paste after filtrate decompression concentrates, standby; The extract clear paste adds acceptable excipient in the pharmacy, and mix homogeneously prepares required drug unit dosage form with the preparation routine techniques.
In the preferred embodiment of the present invention, provided the concrete method for preparing pharmaceutical composition active component of the present invention, this method comprises: (a) take by weighing Radix Scutellariae, Rhizoma Chuanxiong and the Radix Angelicae Sinensis of recipe quantity, be ground into coarse powder; (b) get Radix Scutellariae, add the ethanol extraction of water or 30-80%, filter, filtrate concentrate clear paste I; (c) get Rhizoma Chuanxiong, Radix Angelicae Sinensis, mix, add the water distillation, collect volatile oil, standby; Medicinal residues gained decocting liquid filters, and filtrate concentrates, and gets clear paste II; (d) with clear paste I, clear paste II and the volatile oil mix homogeneously of (b), the preparation of (c) step, add the pharmaceutically acceptable excipient that flies, with the required unit dosage form of conventional formulation prepared.
In the preferred embodiment more of the present invention, provided the method that another kind of approach prepares pharmaceutical composition active component of the present invention, specifically comprised: (a) take by weighing Radix Scutellariae, Rhizoma Chuanxiong and the Radix Angelicae Sinensis of recipe quantity, be ground into coarse powder; (b) get Radix Scutellariae, Rhizoma Chuanxiong, Radix Angelicae Sinensis, mix, decoct with water distillation, collect volatile oil, medicinal residues decocting liquid filters, and filtrate concentrates, and gets clear paste; (c) with gained clear paste and volatile oil mix homogeneously, add pharmaceutically acceptable excipient, prepare required unit dosage form with the conventional formulation technology.
Among the present invention, the used organic solvent that can dissolve each other with water can be to be selected from the methanol that dissolves each other with water, ethanol, the acetone any one, is preferably the ethanol that dissolves each other with water, and more preferably concentration is the ethanol of 30%-80%.
Among the present invention, used extracting method can be any one mode that decocts in extraction, reflux, extract,, immersion extraction, supersound extraction or the percolation extraction, perhaps adopts the combination of Different Extraction Method.
Among the present invention, acceptable excipient in the pharmacy, it is usual excipients well-known to those skilled in the art, according to the needs of making different preparations, those skilled in the art can make a choice from these conventional excipient, being prepared into required unit dosage form, selected excipient is including, but not limited to filler, disintegrating agent, binding agent, lubricant, antiseptic etc.
In an embodiment of the present invention, the pharmaceutical composition of the present invention that has provided different proportion is used for the anti-experimentation of intending the dementia mice model, thereby draws the ratio range of the comparatively preferred crude drug of crude drug compositions of the present invention.
In other embodiment of the present invention, the extract component that has provided the more excellent proportioning of primary raw material medicine of the present invention be used for anti-scopolamine cause intend dementia mice, anti-angiogenic dementia model mice, to the soft cerebral tissue circulation of mice pharmacological evaluation, prove pharmaceutical composition of the present invention can be used for the being correlated with treatment of central nervous system's degeneration.
Among the present invention, the form of pharmaceutical composition can also be to extract single active ingredient to carry out treatment and the alleviation improvement of single-activity composition combination matching to be used for the treatment of nervus centralis degeneration, especially senile dementia, vascular dementia and mild cognitive impairment from Radix Scutellariae, Rhizoma Chuanxiong and Radix Angelicae Sinensis.Among the present invention, the single-activity composition that extracts from primary raw material medicine Radix Scutellariae, Rhizoma Chuanxiong and Radix Angelicae Sinensis is respectively baicalin, ligustilide and ferulic acid, wherein the weight proportion between each active component is baicalin 18-24 part, ligustilide 10-15 part, ferulic acid 3-6 part is preferably 20 parts of baicalins, 12 parts of ligustilides, 4 parts of ferulic acids.
In embodiments of the present invention, give preferred baicalin, ligustilide and ferulic acid pharmacological research to mice dementia and cognitive disorder model.
The specific embodiment
The present invention further illustrates by following embodiment, but any embodiment or its combination not should be understood to the restriction to scope of the present invention or embodiment.Scope of the present invention is limited by appended claims, and in conjunction with the general general knowledge of this description and this area, those of ordinary skills can clearly understand claims institute restricted portion.Under prerequisite without departing from the spirit and scope of the present invention, those skilled in the art can carry out any modification or change to technical scheme of the present invention, and this modification and change are also contained in the scope of the present invention.
The preparation of embodiment 1 granule of the present invention
Prescription: Radix Scutellariae 150g Rhizoma Chuanxiong 150g Radix Angelicae Sinensis 50g
Take by weighing the above-mentioned medicine of recipe quantity, be ground into coarse powder, Radix Angelicae Sinensis, Rhizoma Chuanxiong merge, and add the water of 10 times of weight, soak, and adopt the method for vapor distillation, extract and collect the gained volatile ingredient, and be standby; The water extract filters, and is standby; Filtering residue and Radix Scutellariae coarse powder merge, boil extraction 3 times with the water arrow, filter merging filtrate, gained filtrate merges with Rhizoma Chuanxiong, Radix Angelicae Sinensis water extract again, concentrating under reduced pressure gets clear paste, and the volatile ingredient of gained is joined in the clear paste mix homogeneously, add starch and microcrystalline Cellulose in the clear paste, the system soft material, granulation, dry, granulate promptly get granule.
The preparation of embodiment 2 the present invention tablet
Prescription: Radix Scutellariae 150g Rhizoma Chuanxiong 100g Radix Angelicae Sinensis 100g
Take by weighing the crude drug of recipe quantity, be ground into coarse powder, merge each raw medicinal material, add 70% ethanol of 20 times of amounts, soaked overnight treats that medical material soaks into, adopt percolation to carry out percolation and extract, collect percolate, add fresh 70% ethanol in the percolator in good time and replenish, treat that the percolate color is thin out, percolation extracts and finishes, and merges the percolate of collecting, concentrating under reduced pressure, get crude drug active component extract clear paste, standby; Add starch, microcrystalline Cellulose, cross-linked carboxymethyl cellulose in the extract clear paste, system soft material, granulation, drying, granulate, the gained granule adds magnesium stearate lubricant again, and tabletting promptly gets tablet.
The preparation of embodiment 3 capsules of the present invention
Prescription: Radix Scutellariae 200g Rhizoma Chuanxiong 50g Radix Angelicae Sinensis 50g
Take by weighing the crude drug of recipe quantity, be ground into coarse powder, merge Rhizoma Chuanxiong and Radix Angelicae Sinensis, the water that adds 10 times of amounts is dipped to medical material and soaks into, and extracts with the vapor distillation method and collects the gained volatile ingredient, and is standby; Aqueous extract filters, and gets filtrate, and filtering residue reuse water extraction twice filters, and each time water extract is merged, and concentrating under reduced pressure gets clear paste I;
Radix Scutellariae adds 50% alcohol reflux 3 times, filter, merging filtrate, filtrate decompression concentrates, Radix Scutellariae extracts clear paste II;
Clear paste I, II and volatile ingredient are merged evenly, add dextrin and microcrystalline Cellulose, system soft material, granulation, drying, granulate are packed whole good granule in the hard capsule softgel shell into, promptly get hard capsule.
The preparation of embodiment 4 drop pills of the present invention
Prescription: Radix Scutellariae 50g Rhizoma Chuanxiong 150g Radix Angelicae Sinensis 150g
Get recipe quantity Radix Scutellariae, Rhizoma Chuanxiong, Radix Angelicae Sinensis, be ground into coarse powder, mix homogeneously adds the alcohol reflux 3 times of 10 times of amounts 60% in the medicinal powder, filter, merging filtrate, filtrate decompression concentrates, vacuum drying is pulverized, active component extract medicated powder.
Extract medicated powder according to the preparation technique of this area routine, adds the required excipient of preparation drop pill, makes drop pill.
The preparation of embodiment 5 oral liquids of the present invention
Prescription: Radix Scutellariae 150g Rhizoma Chuanxiong 150g Radix Angelicae Sinensis 150g
Get Radix Scutellariae, Rhizoma Chuanxiong, the Radix Angelicae Sinensis of recipe quantity, be ground into coarse powder, merge, the water that adds 10 times of amounts in the medical material is to soaking into, and vapor distillation extracts, and it is standby to collect volatile ingredient; Extracting liquid filtering, medicinal residues reuse water reflux, extract, twice is filtered, and merges each time aqueous extract, is evaporated to relative density and is about 1.05, adds the volatile ingredient of gained, and adds required antiseptic and the cosolvent of preparation oral liquid, is prepared into oral formulations.
The single extractions preparation of embodiment 6 and the effective ingredient tablet
Prescription: baicalin 2g ligustilide 1.2g ferulic acid 0.4g
Take by weighing baicalin, ligustilide and the ferulic acid of recipe quantity, merge evenly, add filler microcrystalline Cellulose and disintegrating agent crospolyvinylpyrrolidone, alcoholic solution with polyvidone is a binding agent, system soft material, granulation, granulate add magnesium stearate lubricant in the gained granule, and compacting in flakes.
The anti-test of intending the dementia mice model of embodiment 7 drug particles of the present invention
The cholinergic theory thinks that the minimizing of Maynert basal nuclei and Hippocampus and neopallium district cholinergic neuron causes the acetylcholine neurotransmitter level to reduce in the dementia patients brain, thereby causes the obstacle of cognitive function and emotion behavior.Scopolamine is a M cholinoceptor blocker, acetylcholine capable of blocking is to the agonism of m receptor, cause damage in learning and memory, so adopt mouse peritoneal injection scopolamine hydrobromide liquid (SCOP) can cause learning memory disorder, the ability of learning and memory that can simulate the alzheimer disease patient goes down, and can be used for the screening of early stage medicine.Therefore select escape latency in the darkness avoidance test (S) and errors number to carry out Radix Scutellariae, Rhizoma Chuanxiong and Radix Angelicae Sinensis three flavor drug matching proportioning ratio screenings as performance assessment criteria.
1 experiment material
1.1 animal
The KM mice, body weight 18-22g, the SPF level is provided by Sichuan Academy of Medical Sciences institute of lab animals, the animal quality certification number: real moving pipe matter SCXK (river) 2004-16 in river.Mice is raised under the light and shade cycle 12h/12h condition at room temperature 22-24 ℃, all freely drinks water except that administration, modeling and test procedure and ingests.The experiment place is three grades of laboratorys of Chengdu University of Traditional Chinese Medicine of State Administration of Traditional Chinese Medicine herbal pharmacology, numbering: TCM-2009-315.
1.2 medicine
Radix Scutellariae, Rhizoma Chuanxiong, Radix Angelicae Sinensis are all available from Chengdu pharmacy of Tongrentang, identify through doctor Wang Guangzhi of crude drug teaching and research room of Chengdu University of Traditional Chinese Medicine, be the pharmacopeia certified products, according to different dose compatibility proportioning ratios, adopt the method for embodiment 2 to be prepared into 100% standard extracting solution standby (the standard extracting solution is to contain the 1g raw medicinal herbs in every 1ml extracting solution).Scopolamine hydrobromide liquid (SCOP, Shanghai Hefeng Pharmaceutical Co., Ltd. produces).Aricept is defended material (China) pharmaceutcal corporation, Ltd and is produced lot number 080709.
1.3 reagent and instrument
BA-200 keeps away dark tester (and test box) (mice) automatically, produces by Chengdu TME Technology Co., Ltd., and the ACHE test kit, biological engineering institute, lot number 20090606 are built up in Nanjing.
2 experimental techniques
2.1, the orthogonal design of drug ratio
Select L9 (3
4) orthogonal array experimentizes, and sees Table 1.
Table 1 Radix Scutellariae, Rhizoma Chuanxiong and Radix Angelicae Sinensis proportioning ratio (g)
| The experiment number | Radix Scutellariae | Rhizoma Chuanxiong | Radix Angelicae Sinensis |
| ??1 | ??5 | ??5 | ??5 |
| ??2 | ??5 | ??10 | ??10 |
| ??3 | ??5 | ??15 | ??15 |
| ??4 | ??10 | ??5 | ??10 |
| ??5 | ??10 | ??10 | ??15 |
| ??6 | ??10 | ??15 | ??5 |
| The experiment number | Radix Scutellariae | Rhizoma Chuanxiong | Radix Angelicae Sinensis |
| ??7 | ??15 | ??5 | ??15 |
| ??8 | ??15 | ??10 | ??5 |
| ??9 | ??15 | ??15 | ??10 |
2.1 model preparation
Keep away dark experimental group respectively organize mice respectively at administration on the 14th after 50min, except that the blank group, all the other respectively organize mice 10min before keeping away dark training, ip gives SCOP 2mg/kg body weight (0.1ml/10g body weight), causes mouse memory acquired disturbance model.The N.S of blank group ip equivalent.
2.2 grouping and administration
Mice adapts to three days, 120 mices are divided into 12 groups at random, 10/group, i.e. normal control group (A), model group (B), aricept positive controls (C), D (Radix Scutellariae: Rhizoma Chuanxiong: Radix Angelicae Sinensis=1: 1: 1), E (Radix Scutellariae: Rhizoma Chuanxiong: Radix Angelicae Sinensis=1: 2: 2), F (Radix Scutellariae: Rhizoma Chuanxiong: Radix Angelicae Sinensis=1: 3: 3), G (Radix Scutellariae: Rhizoma Chuanxiong: Radix Angelicae Sinensis=2: 1: 2), H (Radix Scutellariae: Rhizoma Chuanxiong: Radix Angelicae Sinensis=2: 2: 3), I (Radix Scutellariae: Rhizoma Chuanxiong: Radix Angelicae Sinensis=2: 3: 1), J (Radix Scutellariae: Rhizoma Chuanxiong: Radix Angelicae Sinensis=3: 1: 3), K (Radix Scutellariae: Rhizoma Chuanxiong: Radix Angelicae Sinensis=3: 2: 1), L (Radix Scutellariae: Rhizoma Chuanxiong: Radix Angelicae Sinensis=3: 3: 2); Press 0.2ml/10g body weight/day ig administration, model control group, blank group give the equivalent normal saline, and aricept group dosage is the 1.3mg/kg body weight; Administration 15 days begins to carry out the test of learning and memory behavioristics after administration on the the 14th, 15.
2.3 ability of learning and memory in mice test (darkness avoidance test)
2.3.1 keep away dark training: keep away dark test group mice behind experiment ig administration on the 14th or distilled water 50min, except that the blank group, all the other respectively organize ip in mice .SCOP modeling, behind the 10min animal back put into the hole after the modeling and keep away the bright chamber of camera bellows, adapt to 3min, logical 36V alternating current, timing at once, incubation period and the errors number of each animal in the record 5min.
2.3.2 keep away dark test: keep away dark group of mice behind experiment (keeping away dark training after 24 hours) ig administration on the 15th or N.S 60min, the same method is put into mice and is kept away the bright chamber of camera bellows, logical simultaneously 36V alternating current, timing, incubation period and the errors number of each animal in the record 5min.
2.4 statistical method
The data " mean ± standard deviation " (
) expression, utilization SPSS11.5 statistical software carries out data statistic analysis.
3 results
3.1 medicine is to intending the influence of dementia mice model darkness avoidance test escape latency and errors number
Compare with the normal control group, model control group mice escape latency obviously shortens, and errors number significantly increases, and significant statistical significance (P＜0.01) is arranged.Compare Radix Scutellariae: Rhizoma Chuanxiong with model control group: different proportioning side's groups of Radix Angelicae Sinensis and positive drug escape latency obviously prolong, and errors number significantly reduces, and tangible statistical significance (P＜0.05) is arranged.See Table 2.
Each proportioning of table 2 is formed the influence (darkness avoidance test) of medicine to mice escape latency and errors number
Annotate:
*: compare P＜0.05 with model group;
*: compare P＜0.01. with model group
3.2 Radix Scutellariae: Rhizoma Chuanxiong: Radix Angelicae Sinensis dose compatibility ratio variance analysis
3.2.1 with the escape latency is index
Table 3 Radix Scutellariae: Rhizoma Chuanxiong: Radix Angelicae Sinensis dose compatibility ratio analysis of variance table (escape latency)
With the escape latency is to investigate index, shows that by extreme difference size in the table it is B＞A＞C that each influence factor acts on primary and secondary; The results of analysis of variance shows: A, B factor have significance meaning (P＜0.05), be that Radix Scutellariae, Rhizoma Chuanxiong consumption are major influence factors, the C factor does not have significance meaning (P＞0.05), be that the influence of Radix Angelicae Sinensis consumption is less, so it is good that proportion compatibility is considered with A3B3C1, i.e. Radix Scutellariae: Rhizoma Chuanxiong: the final weight proportion compatibility of Radix Angelicae Sinensis is 15: 15: 5.
3.2.1 with the errors number is index
Table 4 Radix Scutellariae: Rhizoma Chuanxiong: Radix Angelicae Sinensis dose compatibility ratio analysis of variance table (errors number)
With the errors number is to investigate index, shows that by extreme difference size in the table it is A＞B＞C that each influence factor acts on primary and secondary; The results of analysis of variance shows: the A factor has significance meaning (P＜0.05), be that the Radix Scutellariae consumption is a major influence factors, B and C factor do not have significance meaning (P＞0.05), promptly, Rhizoma Chuanxiong, Radix Angelicae Sinensis consumption influence less, so it is good that proportion compatibility is considered with A3B1C1, i.e. Radix Scutellariae: Rhizoma Chuanxiong: the final weight proportion compatibility of Radix Angelicae Sinensis is 15: 5: 5.
4, conclusion
The factor of comprehensive escape latency and errors number two aspects, the Rhizoma Chuanxiong consumption is a major influence factors to escape latency, but to mistake this moment be not major influence factors, so its consumption can be selected suboptimum level, i.e. B2; Though C is not a major influence factors, consumption can be selected C3, consider simultaneously szechuan lovage rhizome-angelica clinical generally all be as medicine to occurring, therefore, determine that A3B2C2 is best compatibility, i.e. Radix Scutellariae: Rhizoma Chuanxiong: the final weight proportion compatibility of Radix Angelicae Sinensis is 15: 10: 10.
In following pharmacology embodiment, medical material ratio that experimental drug compositions group of the present invention is used and active component preparation method adopt embodiment 2 preparation-obtained active component extracts as the experiment administration medicine.
The anti-scopolamine of embodiment 8 drug particles of the present invention causes the test of intending dementia mice
1 experiment material
1.1 laboratory animal:
60 of KM mices, body weight 20 ± 2g, 45～50 days ages of Mus, male and female half and half provide by Sichuan Academy of Medical Sciences institute of lab animals, and the quality certification number is: SCXK (river) 2004-15, at room temperature 22-24 ℃, raise under the light and shade cycle 12h/12h condition, freely drink water and ingest.
1.2 medicine and reagent:
Aricept is defended material (China) pharmaceutcal corporation, Ltd and is produced lot number 080709.Scopolamine hydrobromide liquid (SCOP), Shanghai Hefeng Pharmaceutical Co., Ltd. produces, lot number 6A18007.ACHE test kit, Nanjing build up biological engineering institute, lot number 20090606.
1.3 experiment key instrument and reagent:
DT-200 mice diving tower tester (and test box), Chengdu TME Technology Co., Ltd.
BA-200 keeps away dark tester (and test box), Chengdu TME Technology Co., Ltd. automatically
2 experimental techniques:
2.1 grouping and processing
The KM mice, male and female half and half, the single-activity that is divided into high, medium and low dosage group of normal control group, model control group, aricept positive controls and medicine of the present invention and embodiment 6 by body weight and sex at random becomes grouping, each treated animal is pressed 0.2ml/10g body weight/day ig administration, and dosage sees Table 1.Keep away dark, diving tower experimental group respectively organize mice respectively at administration on the 14th after 50min, except that the blank group, all the other respectively organize mice 10min before keeping away dark training, diving tower training, and ip gives SCOP 2mg/kg body weight (0.1ml/10g body weight), causes mouse memory acquired disturbance model.The N.S of blank group ip equivalent.Each organizes mice when behavioristics's off-test, gets cerebral tissue at once, and the preparation brain homogenate is respectively organized the full brain AchE of mice activity by AchE test kit step explanation mensuration.
2.2 ability of learning and memory in mice test
(1) darkness avoidance test
1. keep away dark training: keep away dark test group mice behind experiment ig administration on the 14th or distilled water 50min, except that the blank group, all the other respectively organize ip in mice .SCOP modeling, behind the 10min animal back put into the hole after the modeling and keep away the bright chamber of camera bellows, adapt to 3min, logical 36V alternating current, timing at once, incubation period and the errors number of each animal in the record 5min.
2. keep away dark test: keep away dark group of mice behind experiment (keeping away dark training after 24 hours) ig administration on the 15th or N.S 60min, the same method is put into mice and is kept away the bright chamber of camera bellows, logical simultaneously 36V alternating current, timing, incubation period and the errors number of each animal in the record 5min.
(2) diving tower method
1. diving tower training: step down test group mice is behind experiment ig administration on the 14th or N.S 50min, adopt the method identical to make mouse memory acquired disturbance model with keeping away dark experiment, behind the 10min each group mice is put on the diving tower, adapt to 3min, give the 32V alternating current then, at once timing, incubation period and errors number in the record animal 5min.
2. diving tower test: diving tower group mice is in experiment the 15th day (the diving tower training is after 24 hours), and behind ig administration or the N.S 60min, the same method is put in mice on the diving tower, gives 32V alternating current and timing simultaneously, incubation period and errors number in the record animal 5min.
2.3 statistical procedures
Use SPSS 15.0 statistical softwares and carry out the data statistics processing.
3 results
3.1 influence (darkness avoidance test) to mice escape latency and errors number
Table 5 medicine is to the influence (darkness avoidance test) of mice escape latency and errors number
Annotate: compare with model group,
*P＜0.05;
*P＜0.01.
As shown in Table 5, compare the incubation period (p＜0.05) of each dosage group of medicine, aricept group energy significant prolongation mice, and the errors number (p＜0.05) of the interior mice of minimizing 5min with model control group.
3.2 composition of prescription (SZ) is to the influence (diving tower method) of mice escape latency and errors number
Table 6 medicament composing prescription is to the influence (diving tower method) of mice escape latency and errors number
Annotate: compare with model group,
*P＜0.05;
*P＜0.01
As shown in Table 6, compare the incubation period (p＜0.05) of each dosage group of medicine, aricept group energy significant prolongation mice, and the errors number (p＜0.05) of the interior mice of minimizing 5min with model control group.
3.3 the memory acquisition disturbance mouse brain is organized the active influence of AchE
Table 7 medicament composing prescription causes the active influence of AchE in the dysmnesia model mouse brain to SCOP
Annotate: compare with model group,
*P＜0.05;
*P＜0.01.
By seeing Table 7 as can be known, compare with model control group, each dosage group of medicine, aricept group can significantly reduce AchE activity (p＜0.05).
4 conclusions
Medicine of the present invention can significantly improve intends the dementia mice ability of learning and memory due to the scopolamine, reduces cerebral tissue ACHE activity.
The test of the anti-angiogenic dementia model of embodiment 9 drug particles of the present invention
1 experiment material
1.1 laboratory animal:
The SD rat, male and female half and half, the SPF level, 350-500g is provided by Sichuan Academy of Medical Sciences institute of lab animals, and the quality certification number is SCXK (river) 2004-15, at room temperature 22-24 ℃, raises under the light and shade cycle 12h/12h condition, freely drinks water and ingests.
1.2 medicine and reagent:
Nimodipine tablet, the poly-flourish pharmacy in Jiangsu Group Co.,Ltd makes lot number 080218; MDA, MAO test kit, Nanjing build up biological engineering institute, lot number 20091106,20091107.
1.3 experiment key instrument:
The MT-200 water maze, Chengdu TME Technology Co., Ltd.
2 experimental techniques
2.1 model preparation
Rat is adopted the permanent ligation bilateral common carotid arteries of 2-VO method, blank group is only separated bilateral common carotid arteries, not ligation, every rat of postoperative im every day benzylpenicillin sodium for injection 100,000 unit infection, totally 3 days, note moisturizing in the art, postoperative is noted insulation, and postoperative is freely raised after January the animal administration of dividing into groups.
2.2 grouping and administration
50 of modeling rat SD rats, be divided into model control group, nimodipine positive controls, the high, medium and low dosage group of medicine and embodiment 6 prescription groups by the body weight stratified random, other gets 10 with criticizing rat as the blank group, each treated animal is pressed 1ml/100g body weight/day ig administration, dosage sees Table 1, model control group, blank group give the equivalent normal saline, successive administration 15 days, after experiment administration on the 11st, begin to carry out the test of water maze behavioristics, position the navigation experiment in preceding 4 days, went platform to carry out the space search experiment on the 5th day.
2.3Morris water maze test learning and memory in rats ability
It is 4 quadrants that MT-200 water maze pond is divided equally, and platform is positioned at SW quadrant central authorities, 24 ℃ of water temperatures, the water surface is higher than platform 1.5cm, and is with milk to opaque in the water, rat every day in SW quadrant centre position towards the pool wall entry, experimental period is 120s, location navigation in preceding 4 days is write down its incubation period automatically by instrument, and the platform time of staying, platform stop distance, through the platform number of times, if rat is not found platform at 120s, count 120s, stop 10s with hypermnesis on the equal platform of experiment all rats of back; Experiment totally 5 days was removed platform on the 5th day, carried out the space search experiment, and the time is 120s, in the space search, manually writes down its incubation period, and write down it automatically through platform number of times, the platform time of staying, platform stop distance by instrument.The results are shown in Table 8.
2.4 rat hippocampus, Content of MDA, the active mensuration of MAO
After each treated animal learning and memory behavioristics experimental test finished, it was frozen standby to get hippocampal tissue immediately, gets blood and separation of serum, by test kit explanation the carrying out active detection of rat hippocampus, Content of MDA and MAO.The results are shown in Table 9 and table 10.
3 experimental results
3.1 medicine is to the influence of Mus space search ability
Table 8 medicament composing prescription is to the influence of VD rat model space learning memory ability
Annotate: compare with model group,
*P＜0.05;
*P＜0.01.
As shown in Table 8, compare with model control group, each dosage group of medicine can significantly shorten the escape latency (p＜0.01) of rat, and increases process platform number of times (p＜0.05), the platform time of staying (p＜0.05), the platform stop distance (p＜0.05) of rat in the 120s.
3.2 to rat hippocampus tissue, Content of MDA, the active influence of MAO
Table 9 pair rat hippocampus is organized MDA content, the active influence of MAO
Annotate: compare with model group,
*P＜0.05;
*P＜0.01.
Table 10 pair rat blood serum MDA content, the active influence of MAO
Annotate: compare with model group,
*P＜0.05;
*P＜0.01.
By table 9 and table 10 as can be known, with model control group relatively, each dosage group of medicine, nimodipine group can significantly reduce rat hippocampus tissue, Content of MDA, MAO activity (p＜0.01).
4, conclusion
Medicine of the present invention can significantly improve the ability of learning and memory of vascular dementia model, the activity of MDA and MAO in reduction hippocampal tissue and the serum.
Embodiment 10 drug particles of the present invention are tested mice pia mater encephali microcirculation
1 experiment material
1.1 laboratory animal:
60 of KM mices, body weight 20 ± 2g, 45～50 days ages of Mus, male and female half and half provide by Sichuan Academy of Medical Sciences institute of lab animals, and the quality certification number is: SCXK (river) 2004-15, at room temperature 22-24 ℃, raise under the light and shade cycle 12h/12h condition, freely drink water and ingest.
1.2 medicine and reagent:
Nimodipine tablet, the poly-flourish pharmacy in Jiangsu Group Co.,Ltd makes lot number 080218; The noradrenaline bitartrate injection, Shanghai Hefeng Pharmaceutical Co., Ltd. produces, lot number: 0360602;
1.3 experiment key instrument and reagent:
XTW-CTV microcirculation color television microscope, Tyke, Beijing Instr Ltd. produces;
2 experimental techniques
Get 60 of mices, be divided into 5 groups, be i.e. blank group, nimodipine matched group, the high, medium and low dosage group of medicine of the present invention and embodiment 6 prescription proportioning groups, 12 every group.Behind the successive administration 7 days, after the anesthesia of lumbar injection 1% pentobarbital sodium, the ventral decubitus head is fixed, and calvarium center skin T-shape cuts, with the outer 3mm of sagittal suture, 3mm is the center under the coronal suture, raises skull with knife blade, exposes pia mater encephali, form the cranium window of an about 5mm of diameter, add normal saline 20ul, postoperative begins test after stablizing 30min, and entire test keeps 37 ℃ of animal heats.Laboratory animal placed under the microcirculation microscope to select diameter be 30～50um arteriole blood vessel, stablize 30min, survey caliber size, drip 0.1%NA10ul again, with XTW-CTV microcirculation color television microscope by means of the JS micrometer ocular observe continuously drip behind the NA 3, the variation of arteriole blood vessels caliber size during 9min.Microcirculatory color, fluidised form, the every visual field site number that interweaves after observing normal pia mater encephali simultaneously and dripping 0.1%NA.The results are shown in Table 11,12,13,14.
3 experimental results
3.1 medicine is to the influence of mice pia mater encephali arteriole blood vessels caliber
The influence of table 11 pair mice pia mater encephali arteriole blood vessels caliber
Annotate: compare with the blank group,
*P＜0.05,
*P＜0.01
Table 12 pair mice pia mater encephali blood capillary net number purpose influence (the every visual field that interweaves
)
Annotate: compare with the blank group,
*P＜0.05,
*P＜0.01
The influence of table 13 pair mice pia mater encephali blood capillary blood stasis model fluidised form
Annotate: compare with the blank group,
*P＜0.05,
*P＜0.01
The influence of table 14 mice pia mater encephali blood capillary blood stasis model color
Annotate: compare with the blank group,
*P＜0.05,
*P＜0.01
By table 11～14 as seen, high, medium and low each the dosage group of medicine of the present invention all can be improved by the rat's pial local microcirculation obstacle due to the norepinephrine, make arteriole vasodilation, blood flow speeds, every visual field net number that interweaves increases, the blood flow fluidised form is also had some improvement, color is reddened, compared significant difference (P＜0.05, P＜0.01) with the blank group.Each dosage group of medicine of the present invention and positive controls pia mater encephali local microcirculation obstacle recover substantially after 9 fens kinds, and the recovery of blank group is not obvious.
4 conclusions
Medicine of the present invention has the microcirculatory effect of the meninges of improvement.
In sum, three crude drug Huangs of the present invention, Rhizoma Chuanxiong, Radix Angelicae Sinensis compatibility use, and can be used for treatment and alleviate senile dementia, vascular dementia or all diseases of mild cognitive impairment, have synergistic function.
Claims (11)
1. a pharmaceutical composition is characterized in that, mainly is by crude drug Radix Scutellariae, Rhizoma Chuanxiong with when being grouped into.
2. pharmaceutical composition according to claim 1 is characterized in that, described crude drug weight proportion is Radix Scutellariae 5-20 part, Rhizoma Chuanxiong 5-15 part, Radix Angelicae Sinensis 5-15 part.
3. pharmaceutical composition according to claim 1 is characterized in that described crude drug weight proportion is Radix Scutellariae 10-15 part, Rhizoma Chuanxiong 8-12 part, Radix Angelicae Sinensis 8-12 part.
4. pharmaceutical composition according to claim 1 is characterized in that described crude drug weight proportion is Radix Scutellariae 12-15 part, Rhizoma Chuanxiong 10-12 part, Radix Angelicae Sinensis 10-12 part.
5. a method for preparing the described active ingredient in pharmaceutical of any claim among the claim 1-4 is characterized in that Rhizoma Chuanxiong and Radix Angelicae Sinensis volatile ingredient are obtained in preparation earlier, and is standby; Rhizoma Chuanxiong, Radix Angelicae Sinensis and Radix Scutellariae clear paste mix homogeneously behind gained volatile ingredient and the organic solvent extraction that dissolves each other through water or with water, promptly get the pharmaceutical composition active component, mixed active component adds acceptable excipient in the pharmacy, prepares required drug unit dosage form with the preparation routine techniques.
6. method for preparing the described active ingredient in pharmaceutical of any claim among the claim 1-4, it is characterized in that getting the crude drug of recipe quantity, after pulverizing merging, the organic solvent extraction that dissolves each other with water or with water 1-4 time, filter, merge each time and extract filtrate, the active component extract clear paste after filtrate decompression concentrates, standby; The extract clear paste adds acceptable excipient in the pharmacy, and mix homogeneously prepares required drug unit dosage form with the preparation routine techniques.
7. according to claim 5 or 6 described methods, it is characterized in that the described organic solvent that dissolves each other with water is any one or two kinds in methanol, ethanol, the acetone; Described extraction can be any one mode or its combination that decocts in extraction, reflux, extract,, immersion extraction, supersound extraction or the percolation extraction.
8. method according to claim 6 is characterized in that the described organic solvent that dissolves each other with water is the ethanol of 30%-80%.
9. pharmaceutical composition according to claim 1, it is characterized in that described crude drug is the single-activity composition that extracts from Radix Scutellariae, Rhizoma Chuanxiong and Radix Angelicae Sinensis, it is respectively the baicalin in the Radix Scutellariae, ligustilide in the Rhizoma Chuanxiong and the ferulic acid in the Radix Angelicae Sinensis, the weight ratio of wherein said baicalin, ligustilide and ferulic acid is baicalin 18-24 part, ligustilide 10-15 part, ferulic acid 3-6 part.
10. the described compositions of any claim has treatment and alleviates application in central nervous system's degeneration medicine in preparation in claim 1-4 or the claim 9.
11. application according to claim 10 is characterized in that described central nervous system's degenerative disease becomes senile dementia, vascular dementia or mild cognitive impairment disease.
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| CN104825933A (en) * | 2015-05-27 | 2015-08-12 | 苗怡文 | Traditional Chinese medicine preparation for treating paralysis and method for manufacturing traditional Chinese medicine preparation |
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