CN103961621A - Medicine composition for treating alzheimer's disease as well as preparation method and application thereof - Google Patents
Medicine composition for treating alzheimer's disease as well as preparation method and application thereof Download PDFInfo
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Abstract
The invention provides a medicine composition for treating alzheimer's disease. The medicine composition is prepared from the following raw material medicines: alpinia oxyphylla miq, dodder, milkwort, dried rehmannia root, schisandra, chuanxiong rhizome, and cape jasmine fruit. The invention further provides a preparation method and application of the medicine composition. The medicine composition is used for treating alzheimer's disease, the effect of the medicine composition is obvious, the quality is stable and controllable, and a new choice is provided for clinical trials.
Description
Technical field
The present invention relates to a kind of pharmaceutical composition for the treatment of Alzheimer, belong to drug world.
Background technology
Since the nineties in 20th century, the dull-witted clinicopathological study of delivering successively confirms, the over-65s old people of 5-10% suffers from certain cognitive decline or dementia, wherein surpass 50% case reason and be Alzheimer (Alzheimer ' s disease, AD), and women more than male.Alzheimer, claims again Alzheimer, be a kind of the geratic period occur take the central nervous system degenerative disease that progressive dementia is principal character.In 1907, by German doctor Alois Alzheimer, described first.Its main clinical manifestation is carrying out property cognitive dysfunction and hypomnesis, personality and behavior change, and confusion of consciousness, thinking and judgment are lost, and self care ability is lost, finally dead.This destructive brain degenerative disease has been deprived quality---the ability of memory, reasoning, abstract and language of the tool human characteristic of victim.World Alzheimer association has delivered famous " the non-research common recognition of Dare " in December, 2005 on < < lancet > >, point out: the whole world has 2,430 ten thousand people to suffer from dementia, every year newly-increased dull-witted case 4,600,000 (every 7 seconds newly-increased 1 routine patients).Number of patients increases by 1 times in every 20 years, and to the year two thousand forty, global dull-witted case will reach 8,110 ten thousand.AD is carrying out property of a kind of central nervous system (CNS) degenerative disease, its characteristic pathological change is senile plaque (Senile plaques, SPs), neurofibrillary tangle (Neurofibrillary tangles, and regional nerve cell death (neuron and nerve synapse are lost extremely) etc. NFTs).NFTs is that in brain neuron, the Tau protein aggregation of Abnormal Phosphorylation forms, and it is how many relevant with the order of severity of AD symptom.Since oneth century, many scientists have carried out tireless exploration and research with regard to the definite cause of disease and the pathogenesis of AD, but its paathogenic factor is complicated, it be unclear that so far.Think at present, AD is a kind of pathological process being caused by multifactor combineds effect such as genetics factor, environmental factors and metabolic factors.Therefore, AD there is no so far specially good effect, can end and effective therapy of reverse disease progress.At present treatment is mainly to improve the significant performance of patient---dysmnesia and affect the symptom etc. of daily life.
Mild cognitive impairment (mildcognitiveimpairment, MCI) is the concept that aging and dull-witted research field proposes recently, is considered to a kind of transitive state between normal aging people Ahl tribulus sea silent sickness (AD) or other dementias.Research and understanding MCI will contribute to AD or other dementias to take effectively prevention and treatment measure, delay developing of disease.
Summary of the invention
Technical scheme of the present invention has been to provide a kind of pharmaceutical composition for the treatment of Alzheimer.Another technical scheme of the present invention has been to provide the preparation method of this pharmaceutical composition.
The invention provides a kind of pharmaceutical composition for the treatment of Alzheimer, the preparation that the crude drug that it contains following weight proportioning is prepared from:
Fructus Alpiniae Oxyphyllae 25-75 part, Semen Cuscutae 50-150 part, Radix Polygalae 50-150 part, Radix Rehmanniae 50-150 part, Fructus Schisandrae Chinensis 15-45 part, Rhizoma Chuanxiong 50-150 part, Fructus Gardeniae 50-150 part.
Further preferably, it is the preparation that the crude drug by following weight proportioning is prepared from:
Fructus Alpiniae Oxyphyllae 25-75 part, Semen Cuscutae 50-150 part, Radix Polygalae 50-150 part, Radix Rehmanniae 50-150 part, Fructus Schisandrae Chinensis 15-45 part, Rhizoma Chuanxiong 50-150 part, Fructus Gardeniae 50-150 part.
Still more preferably, it is the preparation that the crude drug by following weight proportioning is prepared from:
50 parts of Fructus Alpiniae Oxyphyllaes, 100 parts of Semen Cuscutae, 100 parts of Radix Polygalaes, 100 parts of Radix Rehmanniae, 30 parts of Fructus Schisandrae Chinensis, 100 parts of Rhizoma Chuanxiongs, 100 parts of Fructus Gardeniaes.
Wherein, described Semen Cuscutae is that wine boils Semen Cuscutae.
Pharmaceutical composition of the present invention is to be active component by the primary crude drug of Fructus Alpiniae Oxyphyllae, Semen Cuscutae, Radix Polygalae, Radix Rehmanniae, Fructus Schisandrae Chinensis, Rhizoma Chuanxiong, Fructus Gardeniae or water or extractive with organic solvent, the preparation that adds pharmaceutically acceptable adjuvant or complementary composition to be prepared from.
Wherein, described preparation is tablet, capsule, pill, granule, oral liquid.
The present invention also provides a kind of method of preparing described pharmaceutical composition, and it comprises the steps:
A, take the crude drug of each weight proportion;
B, get Fructus Alpiniae Oxyphyllae, Rhizoma Chuanxiong, soak, extract volatile oil, collect volatile oil and extracting solution, stand-by, medicinal residues are standby;
C, get residue medical material, with the medicinal residues of b step, mix, decoct with water, merge extracting solution prepared by b step, add ethanol precipitation, standing, filter, filtrate is concentrated, dry;
D, get desciccate prepared by c step, add the volatile oil of b step and pharmaceutically acceptable adjuvant or complementary composition to be prepared into pharmaceutically conventional preparation.
Wherein, b step is: get Fructus Alpiniae Oxyphyllae and add 30 times of water gagings to soak after 1 hour, add Rhizoma Chuanxiong, by steam distillation, extract volatile oil 4 hours, collect respectively volatile oil and extracting solution, and stand-by; Concentration of alcohol described in c step is: 65%-95% ethanol; Preferably, concentration of alcohol is: 85% ethanol.
The present invention also provides described pharmaceutical composition to prevent or/and the purposes in the medicine for the treatment of Alzheimer in preparation.
The present invention also provides described pharmaceutical composition preparing kidney tonifying, essence replenishing, mind tranquilizing and the heart calming, the purposes in the medicine of the Fructus Alpiniae Oxyphyllae of having one's ideas straightened out.
The present invention also provides the purposes of this pharmaceutical composition in the medicine of preparation treatment mild cognitive impairment.
Medicine material prescription of the present invention, with Fructus Alpiniae Oxyphyllae, Fructus Schisandrae Chinensis tonifying the kidney for tranquilization, is aided with Semen Cuscutae, Radix Polygalae, Radix Rehmanniae kidney tonifying, essence replenishing, holds concurrently with Rhizoma Chuanxiong blood-activating and qi-promoting, and the logical three warmers that rush down of Fructus Gardeniae.Full side's compatibility is used for the treatment of Alzheimer, reaches kidney tonifying, essence replenishing, mind tranquilizing and the heart calming, and the effect of benefiting intelligence of having one's ideas straightened out, drug effect is obvious, and steady quality is controlled, for clinical, provides a kind of new selection.
Accompanying drawing explanation
Fig. 1 medicine oral liquid chromatogram of the present invention
Fig. 2 reference substance chromatogram
The specific embodiment
The preparation of embodiment 1 medicine oral liquid of the present invention
Weighting raw materials: Fructus Alpiniae Oxyphyllae 50g, Semen Cuscutae (wine boils) 100g, Radix Polygalae 100g, Radix Rehmanniae 100g, Fructus Schisandrae Chinensis 30g, Rhizoma Chuanxiong 100g, Fructus Gardeniae 100g; Make 1000ml
Above seven tastes, get Fructus Alpiniae Oxyphyllae and add 30 times of water gagings to soak after 1 hour, add Rhizoma Chuanxiong, by steam distillation, extract volatile oil 4 hours, collect respectively volatile oil and extracting solution, stand-by; Medicinal residues and other medical materials merge, add 10 and enter a times water gaging, extract 2 times, each extraction time 2h, merges all extracting solution, and (temperature is no more than 80 ℃ to concentrating under reduced pressure, vacuum is-0.04-0.08MPa) to proportion, be 1.15-1.20, add ethanol, making it contain alcohol amount is 85%, standing 24 hours, filter, get filtrate, concentrating under reduced pressure (temperature is no more than 80 ℃, and vacuum is-0.04-0.08MPa) is to relative proportion 1.25-1.30, (temperature is no more than 80 ℃ to drying under reduced pressure, vacuum is-0.04-0.08MPa), pulverize, obtain.
Get purified water 1000ml, be heated to boiling, slowly add intermediate, limit edged stirs, and stirs 10-15min after adding again, and is chilled to room temperature, and cold preservation (0-4 ℃) is spent the night.Filter, get filtrate, get filtrate and add 4g steviosin to be stirred to dissolving completely, get 200mL filtrate, be heated to 70 ℃, add the about 4ml of tween 80, stir, be cooled to 40-50 ℃, add volatile oil, stir 5min, by remaining 800ml medicine liquid heating to 20-30 ℃, the filtrate that has added Tween 80 is slowly added in remaining filtrate, limit edged stirs, and adds and stirs 5min again again, be chilled to room temperature, cold preservation 12 hours, filters, filtrate, embedding (10ml/ props up), obtains.
This product is oral liquid, and content is that brown color arrives tan supernatant liquid, mildly bitter flavor.
The preparation of embodiment 2 medicines of the present invention
Weighting raw materials: Fructus Alpiniae Oxyphyllae 25g, Semen Cuscutae (wine boils) 50g, Radix Polygalae 50g, Radix Rehmanniae 50g, Fructus Schisandrae Chinensis 15g, Rhizoma Chuanxiong 50g, Fructus Gardeniae 50g, decocts with water, concentrated, adds correctives etc. to be prepared into oral liquid.
The preparation of embodiment 3 medicines of the present invention
Weighting raw materials: Fructus Alpiniae Oxyphyllae 75g, Semen Cuscutae 150g, Radix Polygalae 150g, Radix Rehmanniae 150g, Fructus Schisandrae Chinensis 45g, Rhizoma Chuanxiong 150g, Fructus Gardeniae 150g, directly beat powder, tabletting, obtains tablet.
The preparation of embodiment 4 medicines of the present invention
Weighting raw materials: Fructus Alpiniae Oxyphyllae 75g, Semen Cuscutae 50g, Radix Polygalae 50g, Radix Rehmanniae 150g, Fructus Schisandrae Chinensis 45g, Rhizoma Chuanxiong 50g, Fructus Gardeniae 50g, add 75% ethanol extraction, collect alcohol extract, concentrated, extractum adds starch, granulates, encapsulated.
Embodiment 5 medicine material Study on extraction of the present invention
1, the research of volatile oil extracting parameter in medical material
Take respectively Fructus Alpiniae Oxyphyllae 50g, two parts of 75g, pour in flask and are placed in electric jacket, and numbering; In No. 1 flask, add 1500ml water, in No. 2 flasks, add 2250ml water.Soak and add respectively Rhizoma Chuanxiong 100g, 150g after 1 hour.Connect condensing tube and volatile oil extractor starts heating extraction, collect volatile oil.And the amount of writing time and volatile oil collection, experimental result is as table 1
Table 1 volatile oil collecting amount over time
Experimental result shows, due to the increase along with the time, volatile oil extracted amount first increases, and since four hour, along with the increase of time, the amount of volatile oil no longer increased always, therefore Best Times is extracted in Fructus Alpiniae Oxyphyllae, Rhizoma Chuanxiong, is 4 hours.
2 confirmatory experiments
Divide another name Fructus Alpiniae Oxyphyllae 50g, tri-parts of 75g, pour in flask and are placed in electric jacket, and numbering.In No. 1 flask, add 1500ml water, in No. 2 flasks, add 1500ml water, in No. 3 flasks, add 2250ml water.Soak and after one hour, add respectively Rhizoma Chuanxiong 100g, 100g, 150g.Connect condensing tube and volatile oil extractor and start heating extraction 4 hours, collect volatile oil.Experimental result is as following table 2:
Table 2 demonstration test volatile oil is collected result
As can be known from the above table, this prescription Chinese crude drug volatile oil extracts 4 hours, volatile oil can be extracted completely, and extraction process is stable, and repeatability better.
3 water extraction process parameter study
Determining and investigation index of 3.1 factor levels
Crude drug in medicine material prescription of the present invention is taken as main technique route with water extraction, and its technological parameter is studied.
From document, the influence factor of extracting technique of Chinese medicine has a lot, the solvent amount that adds of the main research of this experiment medical material, extraction time, extraction time and extraction solvent.Take water as extracting solvent, and choosing the solvent amount (A) that adds, extraction time (B), extraction time (C) is investigation factor, take Determination of Gardenoside and be to investigate index containing thing amount admittedly, adopts L
9(3
4) orthogonal table arrangement test.The preferred factor level of extraction process is in Table 3.
Table 3 extraction process factor level table
3.2 test methods and result
Take respectively 9 parts of medical materials, in every part containing Semen Cuscutae 100g, Radix Polygalae 100g, Fructus Schisandrae Chinensis 30g, Radix Rehmanniae 100g, Fructus Gardeniae 100g.Medicinal residues with extracting after Fructus Alpiniae Oxyphyllae 50g, Rhizoma Chuanxiong 100g4 hour, arrange test by table 4.Experiment finishes rear collection extracting solution, mixes and measure admittedly to measure containing thing and Determination of Gardenoside.
3.2.1 Determination of Gardenoside is measured
The preparation of reference substance: accurately weighed jasminoidin standard substance
The preparation of sample: get for examination purely, add appropriate solubilizer, standardize solution, ultrasonicly makes to dissolve completely and mix, standby.
Mobile phase: C
2h
3n:H
2o=15:85 flow velocity: 1ml/min detects wavelength: 238nm;
Computing formula,
Wherein, A
1: sample peak area (mv, sec)
C
o: reference substance concentration (mg/ml)
A
0: reference substance peak area (mv, sec)
V
always: extracting solution cumulative volume (ml)
3.2.2 admittedly containing thing, measure
Get appropriate extracting solution in evaporation at constant temperature ware, on water-bath, steam to dry paste, then be dried 3 hours in 105 ℃ of baking ovens.
This result of the test adopts the method for weighted scoring to evaluate.Because Determination of Gardenoside is leading indicator, admittedly containing thing for this reason, so the shared weight proportion of this test determination Determination of Gardenoside is 70, admittedly be 30 containing the shared weight of thing.
Experimental result is as shown in table 4 and table 5.
Known by analysis of variance table, each factor impact is sequentially A > B > C, wherein carries amount of water total weight is had a significant impact, and other factors is not remarkable to total weight significant difference.According to orthogonal test intuitive analysis, the optimum extraction process of determining medical material is A
3b
3c
3, but consider that B factor significant difference is not remarkable, experimental period is long, therefore selection optimum process condition is A
3b
3c
3, add 10 times of water gagings, extract each extraction time 2h 2 times.
3.2.3 proving test
The medical material amount used by orthogonal test takes medical material, by optimum extraction process parameter, adds 10 times of water gagings, extracts 2 times, and each extraction time 2h, carries out repeated authentication test, and experimental result is as shown in table 6.
Confirmatory experiment result shows, this Extraction technique is comparatively stable, can be used as the extraction process of oral liquid.
The arrangement of table 4 orthogonal test and result
Table 5 orthogonal test analysis of variance table
Table 6 proving test result
4 process for refining researchs
4.1 instruments and reagent
Orthogonal test extracting solution: lot number 20121019
Demonstration test extracting solution: lot number 20121101
Ethanol (95%): analytical pure AR, Chengdu Ke Long chemical reagent factory, lot number 20110823
95% ethanol: Kingsoft, Chengdu chemical reagent company limited, lot number 20120229
Analytical electron balance: FA2004 type, the flat instrument and meter of upper current chart company limited produces
Electronic balance: BS-6KH type, Shanghai Yousheng Balance Co., Ltd. produces
Circulating water type vacuum pump: SHZ-D (III) type, Ke Wei Yongxing, Beijing Instrument Ltd. produces
Electric-heated thermostatic water bath: HH-S6 type, Ke Wei Yongxing, Beijing Instrument Ltd. produces
Vacuum drying oven: DZF-6050 type, Beijing Zhong Xing great achievement Instrument Ltd. produces
Alcohol meter: Wuqiang County, the Hebei province instrucment and meter plant that shines together produces
Ultrasonic cleaner: KQ-100 type, Kunshan Ultrasonic Instruments Co., Ltd. produces
Rotating thin film evaporimeter: RE-52B type, Shanghai Yarong Biochemical Instrument Plant produces
Beaker, graduated cylinder, constant weight evaporating dish, condensing tube, special pH test paper (0.5～5.0), PH reagent paper (0～14), funnel, exsiccator, pipet, glue head dropper, Glass rod, separatory funnel, filter paper, densimeter, iron stand, Crucible tongs, ceramic whiteware dish etc.
Volatile oil is separated: owing to containing more water in the volatile oil of collecting, and volatile oil with shipwreck with separated, therefore the mixed solution of volatile oil and water (about 400ml) is placed in to separatory funnel, add about 140ml petroleum ether, standing, water is separated, then petroleum ether-mixture of volatile oil is put into evaporating dish, on water-bath, (55 ℃) steam extremely without petroleum ether.Obtain the about 3ml of volatile oil.
4.2 methods and result
4.2.1 the processing of concentrated solution and alcohol deposit fluid
4.2.1.1 in concentrated solution, Determination of Gardenoside is measured
By concentrated solution supersound process 30 minutes, precision took the about 0.5g of concentrated solution, added appropriate ultrasonic the making of water and dissolved, and was settled to 100ml.Sample thief filters in right amount, by the syringe draw solution of 1ml, it is expelled in sample injection bottle by microporous filter membrane, with high performance liquid chromatogram, measures, and calculates Determination of Gardenoside.
4.2.1.2 the rate of transform is measured
Get alcohol deposit fluid appropriate (can suitably dilute), filter, by the syringe draw solution of 1ml, it is expelled in sample injection bottle by microporous filter membrane, with high performance liquid chromatogram, measure, calculate Determination of Gardenoside.
4.2.1.3 precipitate with ethanol is liquid-solid, containing thing, measure
Experimental implementation is all identical with 3.2.4.2.
4.2.1.4 concentrated solution determination of water and precipitate with ethanol impurities removing efficiency are measured
By concentrated solution supersound process 30 minutes, precision took concentrated solution in right amount in constant weight evaporating dish, and following experimental implementation is all identical with 3.2.4.2 from " water-bath is steamed to extractum shape ".
4.2.2 purifying process
4.2.2.1 extracting solution is concentrated
It is 1.20 left and right that orthogonal test extracting solution, demonstration test extracting solution are concentrated into respectively to density, to be cooledly to room temperature, measures density, weighed weight.
4.2.2.2 alcohol precipitation concentration research
(1) precipitate with ethanol of concentrated solution
Get two parts of test concentrated solutions, every part of about 100g, slowly adds 95% appropriate amount of ethanol, and limit edged stirs, and making into concentration of alcohol is 65%, 85%.Standing over night, filters, and measures filtrate volume.Experimental result is as shown in table 7.
(2) the precipitate with ethanol rate of transform is measured
Get each part of orthogonal test alcohol deposit fluid appropriate, experimental result is as shown in table 7.
(3) precipitate with ethanol is liquid-solid measures containing thing and precipitate with ethanol impurities removing efficiency
Get alcohol deposit fluid appropriate, experimental result is as shown in table 7.
The table 7 orthogonal test precipitate with ethanol rate of transform and precipitate with ethanol are liquid-solid containing thing and precipitate with ethanol impurities removing efficiency measurement result
Experimental result shows, when the alcohol precipitation concentration of concentrated solution is 85%, the precipitate with ethanol rate of transform and precipitate with ethanol impurities removing efficiency are the highest.
4.2.2.3 alcohol precipitation concentration replication experiment
Get concentrated solution and slowly add 95% appropriate amount of ethanol, limit edged stirs, and making into concentration of alcohol is 85%.Standing over night, filters, and measures filtrate volume.Experimental result is as shown in table 8.
The demonstration test precipitate with ethanol rate of transform, experimental result is as shown in table 8.
The table 8 demonstration test precipitate with ethanol rate of transform and precipitate with ethanol are liquid-solid containing thing measurement result
Confirmatory experiment result shows, this process for refining parameter is comparatively stable, and the best alcohol precipitation concentration of oral liquid extracting solution is 85%.
4.2.3 alcohol deposit fluid is concentrated and dry
Demonstration test alcohol deposit fluid is concentrated into density 1.25 left and right in Rotary Evaporators, spreads in ceramic whiteware dish, heating in water bath to concentrated solution is extractum shape, in 80 ℃ of drying under reduced pressure approximately 12 hours, weighs after cooling, obtains brown color to brownish red intermediate.
The quality control of embodiment 6 medicine oral liquids of the present invention
This product 10ml is got in [discriminating] (1), adds respectively petroleum ether (60～90 ℃) 10ml extraction 3 times, divides and gets petroleum ether solution, merges, and volatilizes, and residue adds 3ml ethanol solution, makes to dissolve, as need testing solution.Get Fructus Alpiniae Oxyphyllae control medicinal material powder 1.0g, add petroleum ether (30～60 ℃) 20ml, put supersound extraction 50min in supersound extraction device, filter, volatilize, residue adds 1ml ethanol solution to be made to dissolve, in contrast product solution.According to thin layer chromatography (2010 editions one appendix VI B of Chinese Pharmacopoeia), test, draw above-mentioned two kinds of each 10uL of solution, put respectively on the lamellae of same silica GF254, cyclohexane extraction-the ethyl acetate (9:1) of take is developing solvent, launch, take out, dry, put under ultra-violet lamp (253.7nm) and inspect, in test sample chromatograph, on the relevant position with control medicinal material chromatograph, the principal spot of aobvious same color.
(2) get this product 10ml, the 20ml that adds diethyl ether, reflux 1h, gets upper strata diethyl ether solution, volatilizes, and residue adds ethyl acetate 2ml to be made to dissolve, as test liquid.Get Rhizoma Chuanxiong control medicinal material powder 1.0g, the 20ml that adds diethyl ether, reflux 1h, filters, and filtrate volatilizes, and residue adds ethyl acetate 2ml to be made to dissolve, in contrast liquid.According to thin layer chromatography (2010 editions one appendix VI B of Chinese Pharmacopoeia), test, draw above-mentioned two kinds of each 10uL of solution, put respectively on the lamellae of same silica GF254, cyclohexane extraction-the ethyl acetate (3:1) of take is developing solvent, launch, take out, dry, put under ultra-violet lamp (253.7nm) and inspect, in test sample chromatograph, on the relevant position with control medicinal material chromatograph, the principal spot of aobvious same color.
[assay] jasminoidin is measured according to high performance liquid chromatography (appendix VI D)
Chromatographic condition and system suitability adopt C18 post (4.6mm * 250mm, 5um), and the second cyanogen-water (15:85) of take is mobile phase, and detection wavelength is 238nm, and flow velocity is 1ml/min, and sample size is 10ul.
The preparation precision of reference substance solution takes appropriate jasminoidin, is placed in the brown volumetric flask of 10ml, gets dissolve with methanol solution to scale, makes 1ml containing 0.403mg solution, shakes up, and filters, and obtains.
The preparation precision of need testing solution takes the medicine oral liquid 1ml of the present invention under content uniformity, is placed in 100ml volumetric flask, and water is settled to scale, mixes, and filters, and gets subsequent filtrate, as need testing solution.
Algoscopy is accurate contrast solution and each 10ul of need testing solution of drawing respectively, and injection liquid chromatography, measures, and obtains.The every ml of this product is not less than 11.4mg/ml containing Fructus Gardeniae in jasminoidin.Chromatogram is shown in Fig. 1, Fig. 2
By concrete pharmacodynamics test and clinical trial, prove beneficial effect of the present invention below.
The pharmacological testing of test example 1 medicine oral liquid of the present invention
(1) medicine of the present invention causes the impact of the acquired Disorder Model mice of memory on scopolamine
Along with the prolongation of human longevity, more and more elderly populations are subject to the puzzlement of AD, have a strong impact on patient's quality of life.The mechanism that AD produces is very complicated, completely unclear yet so far, but more and more evidences show, cholinergic nerve of centrum system and cognitive function are closely related.
1 materials and methods
1.1 laboratory animal
60 of animal health Kunming mouses, clean level, male and female half and half, body weight (20 ± 2) g, by Chengdu, Da Shuo bio tech ltd provides, animal production licence number SCXK (river) 2008-24.
Environment mice is fed in clean white plastic mouse cage, 5, every cage.Indoor illumination, humidity, temperature suitable (air-conditioning is controlled 20～26 ℃ of room temperatures, relative humidity 60～70%), ventilation condition is good.
Feedstuff keeps sufficient drinking-water and nutrient fodder.Animal feeding management is carried out according to rule of operation.1.2 medicines and reagent
Medicine of the present invention (is comprised of Fructus Alpiniae Oxyphyllae, Semen Cuscutae, Radix Polygalae, Radix Rehmanniae, Fructus Schisandrae Chinensis, Rhizoma Chuanxiong, Fructus Gardeniae seven taste Chinese medicines, the oral liquid of being prepared by embodiment 1): by Sichuan, Shu Yuan Bo Ye Pharmaceutical Technology Co., Ltd provides, be processed into containing crude drug 1.2g/ml concentrated solution, in experiment, dosage calculates with the amount of contained crude drug in water extraction liquid.Scopolamine hydrobromide, Xuzhou Lai En pharmaceutcal corporation, Ltd.Donepezil hydrochloride, defends material (China) pharmaceutcal corporation, Ltd.
1.3 key instruments and equipment
The long readout instrument of VARIOSKAN FLASH3001 all-wave, U.S. Thermo company, 20PR-5 type High speed refrigerated centrifuge is HIT's product, HH-S type constant water bath box, Germany Sartorius company, WMT-100Morris water maze device and XCS2 mice diving tower instrument are purchased from Chengdu TME Technology Co., Ltd..
2 methods
2.1 animal grouping and administration
Through Morris water maze behavioristics test, reject learning and remembering ability poor or strong, the bad person of swimming position, filter out learning and remembering ability normal mouse.The mice that primary dcreening operation is qualified is divided into 6 groups at random by body weight, sex, is respectively normal blank group, model group, positive control drug group, medicine high dose group of the present invention, middle dosage group, low dose group.Every group of 10 mices, minute cage adaptability is fed 1w.Successive administration 7d before test, according to high dose group (0.76g/ml), middle dosage group (0.38g/ml), low dose group (0.19g/ml), every day, gavage was 1 time.Normal group, model group normal saline gavage.Positive group gives aricept, after gavage 1w, mice is carried out to water maze and the test of diving tower method.
(1) Morris water maze laboratory is in modeling in the 6th day of test, and after except blank group, each group gives lumbar injection scopolamine hydrobromide (3mg/kg), 30min carries out water maze laboratory.
(2) test of diving tower experiment water maze test is the 6th day, after water maze laboratory finishes, mice is carried out to the training of diving tower method, modeling in the 7th day, and except blank group, each group gives lumbar injection scopolamine hydrobromide
(3mg/kg) after, 30min carries out diving tower experiment.
The experiment of 2.2 behavioristicss
2.2.1 water maze laboratory
After administration finishes the 1st day, each is organized mice and starts in Morris water maze, to carry out behavioristics's test, before test, allows Mus free swimming to be familiar with environment.Mice is put into pond from three quadrants in the face of pool wall respectively, record this Mus in 90s and find the platform time used (escape latency).If surpass 90s, also do not find platform, by experimenter, with hands, cause platform, and stop 20s on platform, escape latency is designated as 90s, continuous 6 days, and the 7th day, remove platform, make mice in without platform situation, find the platform in memory, swimming 90s.Record the movement locus of mice, observe mice and pass through the number of times of original platform position, and gather the total time that it rests on original platform quadrant.
2.2.2 diving tower experiment
2 days test periods, training continues to give tested medicine with test period.After administration 1h, start training, animal is put into reaction chamber endoadaptation 3min, passing to immediately 36V alternating current stimulates mice.Then mice is placed on to (timing starts) on insulated platform, record each Mus and jump off the number of times (jumping off the errors number of platform) that in incubation period, 5min of platform, every mice is shocked by electricity for the 1st time, within the 2nd day, test, mice is placed on to (timing starts) on insulated platform, record and jump off the incubation period of platform, the errors number of jumping off platform in 5min for the 1st time, using this as school grade.
3 statistical procedures
Data SPSS17.0 software processes, data represent with x ± s.Between group, mean relatively adopts one factor analysis of variance (ANOVA) check, and P < 0.05 is for there being statistical significance.
4 results
4.1Morris water maze laboratory
4.1.1 orientation navigation experiment
In 3 day training period, each average escape latency of organizing mice is downward trend gradually, shows that the ability of respectively organizing mice study searching platform all increases in all previous learning training.Since the 4th day, model group mice escape latency obviously extended (P ﹤ 0.05), and donepezil hydrochloride positive controls can shorten mice escape latency (P ﹤ 0.05).The high, medium and low dosage group of medicine of the present invention all can shorten mice escape latency (P ﹤ 0.05); In addition the high, medium and low dosage group of medicine of the present invention all can make the spanning platform increased frequency (P ﹤ 0.05) that mice declines and causes due to ability of learning and memory.Between group, compare zero difference (P ﹥ 0.05).(in Table 9)
Table 9 medicine of the present invention on scopolamine cause learning memory disorder model escape latency impact (
n=10)
Note: all with model group comparison,
*p ﹤ 0.05,
*p ﹤ 0.01.Lower same.
4.1.2 space search
Remove after platform, model group mice is many swims around pool wall, longer in the labyrinth outer shroud time of staying, less trip is near platform, its movement locus is randomly dispersed in each quadrant, its spanning platform number of times, effective district's time of staying, effective district's swimming distance, organize obvious minimizing (P < 0.01) with blank.The donepezil hydrochloride positive controls effective coverage time of staying, the effective distance of swimming, relatively extend (P < 0.01) with model group,
The high, medium and low dosage group of medicine of the present invention all can extend the mice effective coverage time of staying, increases effective coverage swimming distance (P < 0.05).Between group, compare zero difference (P ﹥ 0.05) (in Table 10).
Table 10 medicine of the present invention on scopolamine cause learning memory disorder model space search impact (
n=10)
4.2 diving tower
Donepezil hydrochloride matched group can shorten mice incubation period (P < 0.05) to model group mice apparently higher than matched group (P < 0.05) incubation period.Medicine high dose group of the present invention can shorten mice incubation period (P < 0.05); In addition the errors number number of times that in medicine of the present invention, dosage group can make mice cause because ability of learning and memory declines reduces (P < 0.05).Between group, compare zero difference (P ﹥ 0.05).(in Table 11)
Table 11 medicine of the present invention on scopolamine cause learning memory disorder model space search impact (
n=10)
(2) medicine of the present invention causes the impact of AD rat model on A β
A β is the main component of SP, and in brain, A β abnormal deposition is the topmost pathological change of AD.Classical A β toxic action theory thinks, A β is the important step in AD morbidity by solvable state to the conversion of soluble state.The A β of a large amount of depositions has neurotoxicity, finally causes the generation of patient clinical symptom.1 materials and methods
1.1 laboratory animal
Sprague Dawley rat, SPF level, male, 2 monthly ages, body weight (200 ± 20) g, Da Shuo bio tech ltd, Chengdu provides, animal production licence number: SCXK (river) 2008-24.Indoor maintenance 12h illumination, 12h lucifuge circulation is raised, and gives standard feed and drinking water, and to control indoor temperature be (22 ± 1) ℃, relative humidity approximately 40%.
1.2 medicines and reagent
Medicine of the present invention (oral liquid of being prepared by embodiment 1): Shu Yuan Bo Ye Pharmaceutical Technology Co., Ltd provides by Sichuan, is processed into the concentrated solution containing crude drug 1.2g/ml, and in experiment, dosage calculates with the amount of contained crude drug in water extraction liquid.Donepezil hydrochloride, defends material (China) pharmaceutcal corporation, Ltd.
Amyloid fragment (A β
25-35, U.S. Sigma company), before experiment, with physiological saline solution, being diluted to concentration is 2 μ g μ L
-1, in 37 ℃ of calorstats, hatch 72h, formation condensed state is placed on-20 ℃ of refrigerators and deposits standby; Primary antibodie: Anti-A β antibody, Anti-tau antibody (abcam company); β-Actin Antibody (Wuhan doctor's moral is biological); Prestained Protein Marker (Fermentas company); Two anti-and DAB visualizingre agent boxes (the biological company limited of mountain gold bridge in Beijing).
1.3 key instruments and equipment
Labsolution5.5 rat brain stereotaxic instrument (Chengdu TME Technology Co., Ltd.); 5 μ L microsyringes (Town in Shanghai booth injector factory); The electronic dental burr of SAESHIN90+102 (Saeshin PrecisionCo.Ltd); WMT-100Morris water maze video analytic system (Chengdu TME Technology Co., Ltd.); BX43 is just putting microscope (OLYMPUS company); Image-Pro Plus image analysis system (MediaCybernetics, Inc. company); The long readout instrument of VARIOSKAN FLASH3001 all-wave (Thermo company); BSA224S-CW electronic analytical balance (Sartorius company); U.S. Thermo high speed centrifuge (Thermo company).
2 methods
2.1 animal model preparations
According to biliographic data method, each treated animal is all used 10% urethane (10mLkg
-1) after intraperitoneal injection of anesthesia, the flat cranium head of rat position is fixed on brain solid positioner, cut off brain and push up local hair, after routine disinfection, with aseptic operation cutter, at rat scalp median line place, cut one and be about 1.5cm stringer otch, expose bregma, blunt separation subcutaneous tissue and periosteum, according to rat brain stereotaxic atlas, bore and open by aseptic dental burr at 3.4mm, other 2.0mm two places that open, brain median line left and right after skull bregma, with 5 μ L microsyringes, from skull tapping, enter Hippocampus back, rat both sides, depth of needle 2.7mm, slowly at the uniform velocity injects 5 μ L condensed state A β
25-35, injection speed is 1 μ Lmin
-1, 5min injection is complete, and let the acupuncture needle remain at a certain point 2min slowly removes pin, postoperative sewing-up cut, sterilization, sham operated rats is injected isometric normal saline, postoperative freely raising 1 week.
2.2 grouping and administration
60 rat models after modeling are divided into model group, donepezil hydrochloride group (1.02mgkg at random by body weight
-1) and the high (7.2gkg of medicine of the present invention
-1), in (3.6gkg
-1), low (1.8gkg
-1) dosage group.Get 12 rats in sham-operated groups and make blank group, except sham operated rats and model group rat give isopyknic normal saline, each treatment group gives corresponding medicine, and administration volume is 10mLkg
-1, successive administration 30d.
2.3 observation index
2.3.1Morris water maze
Test continuous 5 days, be divided into orientation navigation experiment and space exploration experiment, first 3 days is training period, and the 4th day is orientation navigation experiment, within the 5th day, is space exploration experiment.Setting program is divided into four quadrants by bucket " ten " word, and each quadrant pool wall edge mid-points is place of entry.Platform is placed in fourth quadrant center, and put into water by animal towards pool wall from first, second and third quadrant place of entry every day successively, records animal and finds and climb up platform required time, i.e. escape latency.As do not find platform, incubation period to be recorded as 120s.Within the 5th day, remove platform, from first, second and third quadrant place of entry, animal is put into water towards pool wall successively, record respectively for three times in animal 120s through escape platform number of times, effective district's (2 times of scopes of platform diameter) time of staying, effective district's (2 times of scopes of platform diameter) swimming distance.
2.3.2 pathology HE dyeing
After the test of last behavioristics, by after rat anesthesia perfusion, complete taking-up cerebral tissue, is placed in fixedly 24h of 4% paraformaldehyde, specimens paraffin embedding slices.Dimethylbenzene dewaxing, dyes with hematoxylin, Yihong after dehydrated alcohol dehydration, neutral gum mounting.
2.3.3 SABC
According to SABC test kit explanation, adopt SP two step method carry out routine dewaxing, blocking-up endogenous peroxydase, drip primary antibodie and two anti-, DBA develops the color, haematoxylin is redyed, ethanol dehydration, transparent, neutral gum mounting.Under 400 power microscopes, observe, each of every section measured position and chosen at random 3 visuals field, adopt Image-Pro Plus6.0 image analysis system to analyze, add up every group of positive expression number of targets, the target gross area and accumulative total optical density value (IOD), average and carry out result comparison.The positive is expressed in cytoplasm and/or nucleus for brown color-yellow, and negative cells core is blue.
3 statistical methods
Use SPSS17.0 statistical software to carry out statistical analysis, measurement data with
represent, between many groups, relatively adopt one factor analysis of variance, relatively with least significant difference t check (LSD), do not meet normal distribution and use rank test between two.The P<0.05 of take has statistical significance as difference.
4 results
The experiment of 4.1 behavioristicss
4.1.1 orientation navigation experiment
In 3 day training period, each average escape latency of organizing mice is downward trend gradually, shows that the ability of respectively organizing rat study searching platform all increases in all previous learning training.Since the 4th day, model group rat escape latency obviously extended (P ﹤ 0.05), and donepezil positive controls can shorten rat escape latency (P ﹤ 0.05).The 4th day, high, the middle dosage group of medicine of the present invention all can shorten rat escape latency (P ﹤ 0.05).Between group, compare zero difference (P ﹥ 0.05).(in Table 12)
Table 12 medicine of the present invention on A β cause AD rat model escape latency impact (
n=10)
4.1.2 space search experiment
Remove after platform, model group rat is many swims around pool wall, longer in the labyrinth outer shroud time of staying, less trip is near platform, its movement locus is randomly dispersed in each quadrant, its spanning platform number of times, effective district's time of staying, effective district's swimming distance, with the obvious minimizing of sham operated rats (P < 0.05).The donepezil hydrochloride positive controls effective coverage time of staying, the effective distance of swimming, relatively extend (P < 0.05) with model group,
The high, medium and low dosage group of medicine of the present invention all can extend the mice effective coverage time of staying (P < 0.05), in, low dose group all increases effective coverage swimming distance (P < 0.05).Between group, compare zero difference (P ﹥ 0.05) (in Table 13).
Table 13 medicine of the present invention on A β cause AD rat model space search impact (
n=10)
4.2HE pathological change
In rats in sham-operated group brain, neurocyte is arranged integral body, and a few cell is dense dyes, and part kitchen range neurocyte reduces.In model group rat brain, microglia increases, and neurocyte quantity obviously reduces, and arranges the karyopyknosis of not embarking on journey, part vanished cell, and rarely seen cavity exists.In each treatment group rat brain, neuron pathological changes is all obviously lighter than model group, and in visible part neuron plasma, tiger spot reduces or disappears, and especially obvious with medicine high dose group improvement of the present invention, most of forms are intact, are dispersed in glial cell.
4.3 immunohistochemical methods are observed cerebral tissue Protein tau and are expressed
Under light microscopic, to starch sepia be Protein tau stained positive to observation of cell.Experimental result shows, model control group Hippocampus tau is compared with the remarkable high expressed of sham operated rats, painted darker, and Positive Objects number, Positive Objects area and average optical density value significantly increase (P<0.01).Medication therapy groups of the present invention visible positive staining neuron number and dye levels and sham operated rats are close, compared with model control group, express and reduce, painted more shallow, Positive Objects number, Positive Objects area and average optical density value obviously reduce (P<0.05).(in Table 14)
The impact that table 14 medicine of the present invention is expressed A β rat model Hippocampus tau
(3) conclusion
Medicine of the present invention has certain nootropic effect, can be used for treating Alzheimer.
The clinical trial of test example 2 Drug therapy mild cognitive impairment of the present invention
1. research contents
The impact of 1.1 medicines of the present invention on MCI patient's mental psychology test scale integration
1.2 impacts of medicine of the present invention on MCI patient's tcm syndrome integration
1.3 drug safety evaluation studies of the present invention
2. research approach
2.1 diagnostic criteria
2.1.1 Western medicine diagnose standard
According to the MCI diagnostic criteria of Petereson formulation in 1999 and the diagnostic criteria of DMS-IV, draft diagnostic criteria as follows:
1. patient's readme memory obviously declines, and has family members to confirm;
2. decrease of cognitive function, is mainly that memory function declines, and memory scale detects score at the age with below education coupling matched group 1.5SD,
3. other cognitive functions are as orientation force, judgment, and computing power, space, visions etc. decline not obvious or slightly decline;
4. can not reach dull-witted diagnostic criteria, CDR=0.5;
5. not impact or minimal effect orthobiosis, life can be taken care of oneself;
6. can not explain this cognitive disorder with other diseases of mental and nervous system, as depression, the capable apoplexy of ischemia etc.
2.1.2 TCM syndrome diagnostic criteria
With reference to Zheng Xiao cornel chief editor's < < new Chinese medicine guideline of clinical investigations > > and Zhou Zhongying chief editor's < < Chinese Internal Medicine > >, draft diagnostic criteria as follows:
1. primary symptom: hypophrenia, soreness of the waist and knees, asthenia is thought sleeping, heaviness of the head as if it were tightly bandaged, abundant expectoration salivation, dim complexion or lip onyx are livid purple.
2. time disease: bradyphrenia, vertigo and tinnitus, fat too fat to move, pain in fixed position, eyelid green grass or young crops is black, urinary incontinence.
Possess two of above primary symptoms (hypophrenia indispensable), in addition time disease at least to possess two above persons can clinical syndrome differentiation be shenxu-tanyu card.
2.2 inclusive criteria
1. at 65-80 between year, men and women all can the age;
2. meet MCI diagnostic criteria, and hypomnesis symptom continues more than six months;
3. the rating of clinical dementia rating scale is suspicious dementia (CDR=0.5 divides);
4. the scoring of MMSE scale is between 24 to 27 minutes;
5. Hamilton depressive scale scoring <8 divides;
6. volunteer, and sign Informed Consent Form;
7. there is certain culture degree, previously can read simple newspaper article and write simple little article;
8. drug research meaning is had to correct understanding, the O&A of research worker is had to good compliance.2.3 exclusion standard
1. CDR is assessed as normal or dement;
2. Hamilton depressive scale scoring >8 divides;
3. current patient's spiritedness Split disease, schizoaffective disorder or constitutional adult affective disorder, comprise previous adult's affective disorder medical history person.Or with serious neurologic defect patient, as various aphasias, agnosia person;
4. patient had the diagnosis of AD, VaD, central nervous system infection, wound post-traumatic dementia, poisoning metabolic encephalopathy, HuntingtonShi disease, multiple sclerosis, ParkinsonShi disease before amnesia, and oligophrenia, the untreated endocrinopathy of former are as Qishing or former 's hypothalamic function go down (hypothyroidism of curing not except);
5. merge intentionally, the serious primary disease patient such as brain, liver, kidney and hemopoietic system;
6. there is serious dyspepsia, or Gastrointestinal Obstruction, or the routed patient who swings of harmonization of the stomach duodenum, and the patient who affects other gastroenteropathys of drug absorption;
7. the age is person below 65 years old or more than 80 years old;
8. to this drugs allergy sufferers;
9. during in the recent period once system was accepted, the patient that treats of doctor trained in Western medicine nootropic drug.
2.4 rejecting standards
1. can not complete on request this experimenter;
2. in process of the test, experimenter's compliance is poor, affects effectiveness and safety evaluatio person;
3. this medicine is not tolerated the person that occurs adverse effect.
Coming off and processing of 2.5 cases
The case standard 2.5.1 come off
1. there is serious adverse events, complication and special physiological and change, the person that should not continue reception test or to this institute medicine allergy sufferers;
2. bolter voluntarily in process of the test;
3. because other a variety of causes does not finish to exit test, lost to follow-up or dead case the course for the treatment of;
4. data is incomplete, affects effectiveness and safety judgement person;
The processing of the case that 2.5.2 comes off:
After patient comes off, the contact method that researcher should be filled according to patient, as phone, get in touch with patient in address etc., records last administration time, completes the evaluation item that can complete, and inquire and exit reason, carries out respective record.
Keep properly the correlational study data of the case that comes off, in order to carrying out test statistics analysis.
2.6 test method
2.6.1 case and medicament sources
Observe case from Chengdu front three hospital and Chengdu community hospital.
Medicine of the present invention, is prepared by embodiment 1.
2.6.2 therapeutic scheme
To meeting the object of study of inclusive criteria, do self cross-reference observation, give medicine intelligence of the present invention and exempt to decoct granule, day potion.In therapeutic process, give basis health care support and [comprise four aspects: general health propaganda and education (actively recuperate under medical treatment old complaint, keep happy); Modest movement (take a walk: 1000 meters, each 1 time sooner or later); Limb motion (handball: 30 minutes/day, Bid); General rational nutritional support], do not use other to there is the Chinese and western drugs that expands blood vessel, nootropic effect.Be 12 weeks the course for the treatment of.
2.7 observation index
2.7.1 physical data
1. demography data: comprise age, sex, race, occupation, schooling, height, body weight, medication history and ill history etc.
2. general physical examination: as breathing, heart rate, blood pressure, pulse etc.
2.7.2 health giving quality index
1. mental psychology test scale (table 15) scoring:
The test scale that this institute of table 15 adopts
2. tcm syndrome integral estimation:
Table 16MCI shenxu-tanyu syndrome scalar quantization table
By full-time staff's (measuring consistency check by curative effect through professional training), in quiet environment, tested above.Mental psychology scale is tested before and after treatment and is respectively observed once; Tcm syndrome integration respectively records once for before treatment and after treatment the 1st week, the 4th week, the 8th week, the 12nd week.
2.7.3 safety indexes
Blood, urine, just conventional, hepatic and renal function, electrocardiogram, and a situation arises to record at any time untoward reaction and adverse events, the measure of taking and lapsing to.After within 12 weeks, finishing with treatment before treatment, each detects once.
2.8 curative effect determinate standard
1. mental psychology test scale scoring:
With Clinical Memory scale, must be divided into main reference index, by the test scale score before and after relatively treating, change, evaluate the improvement situation of Function of memory cognition, adopt improvement rate to represent, that is:
Improvement rate=[score before (the front score of score-treatment after treatment)/treatment] * 100%
Effective: improvement rate >=20%;
Effective: 10%≤improvement rate < 20%;
Invalid: improvement rate <10%.
Effective percentage (%)=(effective number+efficiently individual quantity)/total number of persons * 100%
2. tcm syndrome integral estimation:
According to tcm syndrome scalar quantization table, calculate therapeutic index, according to therapeutic index, carry out tcm syndrome therapeutic evaluation.
Therapeutic index (nimodipine method)=[integration before (the rear integration of integration-treatment before treatment)/treatment] * 100%
Clinical recovery: therapeutic index >=80%, symptom, sign disappear or substantially disappear;
Effective: 50%≤therapeutic index < 80%, symptom, sign are obviously improved;
Effective: 20%≤therapeutic index < 50%, symptom, sign all take a turn for the better;
Invalid: therapeutic index < 20%, symptom, sign are not improved or increase the weight of.
Effective percentage (%)=(recovery from illness number+effective number+efficiently individual quantity)/total number of persons * 100%
3. safety evaluatio standard
1 grade: safety, without any untoward reaction.
2 grades: safer, if any untoward reaction, do not need to do any processing and can continue administration.
3 grades: have safety issue, have moderate untoward reaction, after processing, can continue administration.
4 grades: because test is ended in untoward reaction.
2.9 statistical method
All data acquisitions are used " mean ± standard deviation "
represent, with EXCEL worksheet building database and use SPSS19.0 statistics software to carry out statistical test.Measurement data adopts t check, and enumeration data adopts χ
2check.P < 0.05 is difference significance, and P < 0.01 has highly significant meaning for difference.
3 results of study
3.1 come off, reject case
The case that comes off 5 examples.3 examples are lost to follow-up.1 routine diabetes medical history more than 10 years, concurrent retinopathy, because of the operation drug withdrawal of being in hospital.1 example because of the conscious state of an illness lighter, be reluctant to continue to take medicine.
3.2 physical data
Final MCI patient's 85 examples of observing of this research, male 46 examples wherein, female's 39 examples (55～78 years old), 69.23 ± 3.63 years old mean age; The shortest course of disease 0.8 year, the longest 10 years, average course of disease 7.72 ± 2.72 years; 42 people of schooling primary school, the people of junior middle schools 31, senior middle school and above academic 12 people; Dextromanuality 80 people, left handedness 5 people; Physical work 72 people, comprise workman, peasant, soldier etc., and non-manual work 13 people comprise management, accounting, pilot etc.; Sick 45 people of complicated hypertension, coronary heart disease 3 people, hyperlipidemia 43 people, insomnia medical history 30 people, have smoking, history of drinking history 18 people.
3.3 cognitive function curative effect evaluations
After medication 3 months, the overall cognitive level of patient is changed and assessed, respectively MMSE scale (overall cognitive function), Clinical Memory scale (memory), line are detected to (execution function), Boston name scale (linguistic competence), picture clock Survey (visual space structural capacity), activity of daily living ADL scale and GDS scale (scale totally fails) and assess.
1) overall cognitive proficiency assessment
MMSE scale is used for evaluating overall cognitive function, can be divided into the junior units such as orientation force, memory, linguistic competence, but except directional memory, and other are every can not represent the cognitive function in corresponding field separately.
Table 17MMSE scoring
Through Wilcoxon paired test, after treatment, orientation force integration is significantly improved before treating, and bilateral P ﹤ 0.05 is variant before and after the treatment of orientation force integration; MMSE scale total mark bilateral P ﹤ 0.05, has statistical significance.
2) memory ability assessment
Clinical Memory scale, for the main scale of observing of this research, for evaluating memory level, frees recall comprising sensing memory, learning by association, image etc.
The scoring of table 18 Clinical Memory scale
Adopt Wilcoxon paired test, Clinical Memory scale overall score before and after treatment, variant, evident in efficacy before and after bilateral P ﹤ 0.05 explanation Clinical Memory scale total points treatment.Sensing memory, image are freed recall, random shape is re-recognized scoring and all adopted Wilcoxon paired test, and bilateral P ﹤ 0.05 illustrates that sensing memory scoring, image are freed recall, random shape is re-recognized the treatment front and back of marking variant, evident in efficacy.Learning by association, the contact of portrait feature are recalled, and result shows P ﹥ 0.05, no difference of science of statistics.
Trail making test assess patient is carried out function, is divided into A and B two parts, and A partly reflects consciousness movement velocity, and B part is except comprising consciousness movement velocity, also comprises concept and attention switching ability, and to frontal lobe, hypokinetic screening has certain help.
The scoring of table 19 trail making test A-B scale
Before and after treatment, trail making test A and B all adopt paired t-test, A part P ﹤ 0.05, and before and after treatment, there were significant differences, B part P ﹥ 0.05, no difference of science of statistics.
Boston name scale is for detection of patient linguistic competence, and with Wilcoxon paired test, result is shown P ﹥ 0.05, integration no difference of science of statistics before and after treatment.
The scoring of table 20Boston name scale
Draw clock test for detection of patient visual space structural capacity, application Wilcoxon paired test, result is shown P ﹥ 0.05, can not think that treatment front and back integration is variant.
Table 21 is drawn the scoring of clock scale
3.4 tcm syndrome efficacy analysis results
According to syndrome integration method, obtain syndrome integration, and respectively before medication, 12 weeks records respectively after medication.Measurement data all adopts mean ± standard deviation (X ± S) to represent, adopts paired t-test, and result is as shown in the table:
Record gained syndrome total mark respectively with medication before the comparison of syndrome total mark, adopt paired t-test, result shows, treat after 12 weeks and medication is front compares, P ﹤ 0.000, has statistical significance.
3.5 safety evaluatio
This is studied in 85 routine patients, and 1 routine patient merges flu and occurs distending pain in the head after taking medicine one week, with discomforts such as cough, watery nasal discharges.Advise and suspend medicine of the present invention, after 10 days, patient's cold symptoms disappears, and independently continues to take medicine of the present invention, there is not the special discomforts such as distending pain in the head, according to above-mentioned safety evaluatio standard, do not belong to Adverse Event, remaining patient does not all tell period in a medicine any discomfort.
4. conclusion
Drug therapy Alzheimer preliminary stage MCI of the present invention is effective.
Medicine of the present invention is under traditional Chinese medicine theory instructs, progress in conjunction with modern medicine, select AD utmost point commitment as intervening point of penetration, pathogenic characteristic for MCI deficiency in origin and excess in superficiality, employing has tonifying the kidney for tranquilization, by the medicine of the present invention of expectorant dredging collateral effect as medicine, and support in conjunction with basis health care, intend by normalized random contrast clinical trial, utilize a plurality of internationally recognized mental psychology test scales and TCM syndrome evaluation table as health giving quality observation index, large conventional in conjunction with three, Liver and kidney merit, the safety indexes such as electrocardiogram, disclose effectiveness and the safety effect of Drug therapy MCI of the present invention (shenxu-tanyu card).For clinical treatment MCI provides determined curative effect, reliable and stable medicine, to improve the symptoms such as patient's hypophrenia, hypomnesis, bradykinesia, delay or end the process that MCI develops into AD, alleviate the economy, society and the psychological burden that cause thus, improve patient's behavioral competence and living standard.
Claims (12)
1. a pharmaceutical composition for the treatment of Alzheimer, is characterized in that: the preparation that the crude drug that it contains following weight proportioning is prepared from:
Fructus Alpiniae Oxyphyllae 25-75 part, Semen Cuscutae 50-150 part, Radix Polygalae 50-150 part, Radix Rehmanniae 50-150 part, Fructus Schisandrae Chinensis 15-45 part, Rhizoma Chuanxiong 50-150 part, Fructus Gardeniae 50-150 part.
2. pharmaceutical composition according to claim 1, is characterized in that: it is the preparation that the crude drug by following weight proportioning is prepared from:
Fructus Alpiniae Oxyphyllae 25-75 part, Semen Cuscutae 50-150 part, Radix Polygalae 50-150 part, Radix Rehmanniae 50-150 part, Fructus Schisandrae Chinensis 15-45 part, Rhizoma Chuanxiong 50-150 part, Fructus Gardeniae 50-150 part.
3. pharmaceutical composition according to claim 2, is characterized in that: it is the preparation that the crude drug by following weight proportioning is prepared from:
50 parts of Fructus Alpiniae Oxyphyllaes, 100 parts of Semen Cuscutae, 100 parts of Radix Polygalaes, 100 parts of Radix Rehmanniae, 30 parts of Fructus Schisandrae Chinensis, 100 parts of Rhizoma Chuanxiongs, 100 parts of Fructus Gardeniaes.
4. according to the pharmaceutical composition described in claim 1-3 any one, it is characterized in that: described Semen Cuscutae is that wine boils Semen Cuscutae.
5. according to the pharmaceutical composition described in claim 1-3 any one, it is characterized in that: it is to be active component by the primary crude drug of Fructus Alpiniae Oxyphyllae, Semen Cuscutae, Radix Polygalae, Radix Rehmanniae, Fructus Schisandrae Chinensis, Rhizoma Chuanxiong, Fructus Gardeniae or water or extractive with organic solvent, the preparation that adds pharmaceutically acceptable adjuvant or complementary composition to be prepared from.
6. pharmaceutical composition according to claim 5, is characterized in that: described preparation is tablet, capsule, pill, granule, oral liquid.
7. pharmaceutical composition according to claim 6, is characterized in that: in described oral liquid, every ml is not less than 11.4mg/ml containing Fructus Gardeniae in jasminoidin.
8. a method of preparing the pharmaceutical composition described in claim 1-7 any one, it comprises the steps:
A, take the crude drug of each weight proportion;
B, get Fructus Alpiniae Oxyphyllae, Rhizoma Chuanxiong, soak, extract volatile oil, collect volatile oil and extracting solution, stand-by, medicinal residues are standby;
C, get residue medical material, with the medicinal residues of b step, mix, decoct with water, merge extracting solution prepared by b step, add ethanol precipitation, standing, filter, filtrate is concentrated, dry;
D, get desciccate prepared by c step, add the volatile oil of b step and pharmaceutically acceptable adjuvant or complementary composition to be prepared into pharmaceutically conventional preparation.
9. the preparation method of pharmaceutical composition according to claim 8, it is characterized in that: b step is: get Fructus Alpiniae Oxyphyllae and add 30 times of water gagings to soak after 1 hour, add Rhizoma Chuanxiong, by steam distillation, extract volatile oil 4 hours, collect respectively volatile oil and extracting solution, stand-by; Concentration of alcohol described in c step is: 65%-95% ethanol; Preferably, concentration of alcohol is: 85% ethanol.
10. the pharmaceutical composition described in claim 1-7 any one prevents or/and the purposes in the medicine for the treatment of Alzheimer in preparation.
Pharmaceutical composition described in 11. claim 1-7 any one is being prepared kidney tonifying, essence replenishing, mind tranquilizing and the heart calming, the purposes in the medicine of the Fructus Alpiniae Oxyphyllae of having one's ideas straightened out.
The purposes of pharmaceutical composition described in 12. claim 1-7 any one in the medicine of preparation treatment mild cognitive impairment.
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| 杨楠等: "栀子粗提物对拟痴呆模型鼠学习记忆功能障碍的影响", 《中国康复理论与实践》 * |
| 蔡景高等: "健忆口服液改善老年人学习记忆功能的临床与实验研究", 《中国中西医结合杂志》 * |
| 金虹: "治疗老年痴呆中药对乙酰胆碱酯酶的体外抑制作用研究", 《时珍国医国药》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105267615A (en) * | 2015-12-02 | 2016-01-27 | 山东省药学科学院 | Traditional Chinese medicine composition for treating Alzheimer disease and preparation method thereof |
| CN105920237A (en) * | 2016-05-02 | 2016-09-07 | 邹士东 | Pharmaceutical preparation for treating Alzheimer's disease and application of pharmaceutical preparation |
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| CN103961621B (en) | 2017-05-31 |
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