CN109464488A - Honeysuckle general flavone and its preparation method and application - Google Patents
Honeysuckle general flavone and its preparation method and application Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/35—Caprifoliaceae (Honeysuckle family)
- A61K36/355—Lonicera (honeysuckle)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
Abstract
Invention is related to plant pharmaceutical technology field, is related to general flavone and its preparation and the application in preparation treatment Alzheimer disease medicament in honeysuckle general flavone and its preparation method and application more particularly to medicinal plant honeysuckle.The present invention obtains general flavone by being suspended with water after being concentrated under reduced pressure to give crude extract by the method for honeysuckle medicinal part dry flower water extract-alcohol precipitation, with CHP20 column chromatography, and content is greater than the active component in 70% and honeysuckle with pharmacological activity.General flavone in honeysuckle provided by the invention has apparent anti-AD activity, and honeysuckle is integration of drinking and medicinal herbs class plant, without toxic, side effects, can be used as a kind of potential anti-AD drug.The scope of application of honeysuckle is expanded.The method of the present invention design is reasonable, pollution low, general flavone pharmacological activity outstanding feature simple, at low cost with technical process.
Description
Technical field
The present invention relates to plant pharmaceutical technology fields, are related to honeysuckle general flavone and its preparation method and application, especially relate to
And general flavone and its preparation and the application in preparation treatment Alzheimer disease medicament in medicinal plant honeysuckle.
Background technique
Honeysuckle (Lonicera japonica Thunb.) is the dry flower of woodbine honeysuckle, and bud is with fertilizer
Greatly, the clean person that color is bluish white, holds is preferred.Medical raw plant removes Heilungkiang, the Inner Mongol, Ningxia, Qinghai, Xinjiang, Hainan and Tibet
Outer without growth naturally, national each province is distributed.Honeysuckle is a kind of conventional Chinese medicine with long history, first recorded in " name doctor is other
Record ", it is classified as top grade." honeysuckle " one refers under " honeysuckle " item first appeared in Li Shizhen (1518-1593 A.D.) Compendium of Material Medica, has been known as this
The rectification of name of medicinal material, and take in Chinese Pharmacopoeia.Honeysuckle property is sweet cold, and function is clearing heat and detoxicating, anti-inflammatory detumescence, to bacillary dysentery and each
Kind purulent disease is all effective.The honeysuckle preparation produced has " fructus forsythiae antidotal tablet ", " yin huang pian ", " YINHUANG ZHUSHEYE " etc..
" distilled liquid of honeysuckle " is that the armaticity volatile oil that honeysuckle is extracted with the way of distillation and water solubility slip out object, is the non-defective unit of Qinghuo Jiedu,
Infant fetal toxin, boil can be controlled, generated heat the diseases early stage such as thirsty.The research of organic acid is concentrated mainly on for the research of honeysuckle,
The especially research of chlorogenic acid.With going deep into for research, it has been found that the flavone compound in honeysuckle also has excellent
Physiological activity, such as antibacterial, antiviral, anticancer, anti-cancer and inhibition lipase, and it is harmless nontoxic.Therefore, to yellow in honeysuckle
Ketone compounds, which carry out research, has important practical significance.
Summary of the invention
The object of the present invention is to provide general flavone in a kind of plant honeysuckle and preparation method thereof, this method is by honeysuckle
The method of medicinal part dry flower water extract-alcohol precipitation extracts, and is suspended, is passed through with water after being concentrated under reduced pressure to give crude extract
CHP20 column chromatography column finally obtains the higher general flavone of purity.
The present invention is achieved through the following technical solutions:
Honeysuckle general flavone, is prepared via a method which:
(1) traditional Chinese medicine honeysuckle, with water heating and refluxing extraction；
(2) it is concentrated under reduced pressure, industrial alcohol is added in concentrate, filters after standing, recycles ethyl alcohol, obtains medicinal extract；
(3) after adding water to be suspended gained medicinal extract, upper HP20 resin, first with 10%~20% ethyl alcohol water elution, then with 60%
~80% ethyl alcohol water elution collects 60%~80% ethanol water eluent, ethyl alcohol is recovered under reduced pressure, is drying to obtain.
Wherein,
In step (1), first with the water soaked overnight of 2-3 times of volume, then the water heating extracting 2-3 with 5-8 times of volume
It is secondary；
In step (2), the pH value that extracting solution can be adjusted before reduced pressure is 3.00~5.00；
In step (2), aqueous extract is concentrated into 1:1, ethyl alcohol, which is added, makes its alcohol content 70-80%；
In step (3), the HP20 is one or more of CHP20Y, CHP20P, CHP20SS；
In step (3), acid, the acid are as follows: hydrochloric acid can be added in the ethanol eluate.
Resulting extract utilizes FeCl3Color development reaction is positive, and HCl-Mg reaction is positive, and determines that gained mentions
Taking object is general flavone.
Using visible spectrophotometry (one V A of annex of Chinese Pharmacopoeia), with Hyperoside (purity 98%) for reference substance,
And sodium nitrite-aluminum nitrate-sodium hydroxide colorimetric method is used, the red complexing of generation is reacted with aluminium salt according to flavone compound
Object, Detection wavelength 510nm elute the content of obtained general flavone for detecting each chromatographic column.The result shows that with of the invention
The honeysuckle general flavone of method preparation is 70% or more or even 80% or more.
The present invention also provides the honeysuckle general flavone of the method preparation anti-A amyloid beta aggregation activities in vitro
Test, the results showed that, honeysuckle general flavone of the invention have apparent anti-senile dementia effect, can be used for preparing treat Ah
The drug of the silent disease in Wurz sea.
Usefulness of the present invention is: 1, in the case where inhibiting aβ protein aggregation activity to be oriented to isolated guidance, in medicinal material
Active component has carried out sufficient purification, concentration and enrichment, using from previous different extraction process, be that honeysuckle is dry
Bud crushes, and by a step CHP20 resin method after water extract-alcohol precipitation, collection obtains having in its extractive of general flavone and honeysuckle
There is the active component of anti-AD pharmacological activity, the content using spectrophotometry measurement general flavone reaches 70%, is much better than with slightly mentioning
Object is directly used as medicine.2, the general flavone obtained with this method has significant pharmacological action, is able to suppress the aggregation of aβ protein, makes
Rate reaches 60% or more, hence it is evident that higher than the inhibiting rate of reference substance curcumin, and in the water maze of Senile Dementia Model Rats reality
Show good anti-AD activity in testing, 3, honeysuckle as a kind of traditional Chinese medicine, toxicity, side effect is lower, resourceful, right
Alzheimer disease has good treatment potentiality, can develop into a kind of new anti-AD drug.4, the method for the present invention designs
Rationally, have the characteristics that simple process, at low cost, low energy consumption and production process pollute it is low.
Detailed description of the invention
Fig. 1 be using curcumin as positive control, at 20 μM, suppression that general flavone aβ protein that different schemes obtain is assembled
Rate processed, compared with positive control, * indicates that p < 0.05, * * indicate p < 0.01.
Fig. 2 is the obtained general flavone of different schemes to H2O2The neuroprotection of the SH-SY5Y cellular damage of induction.Make
Cytotoxicity of the general flavone (25,50,100 μM) obtained with 5 groups of different schemes of MTT measuring method test in SH-SY5Y cell
Effect.
Fig. 3 is the obtained general flavone of different schemes to H2O2The neuroprotection of the SH-SY5Y cellular damage of induction.?
In the case where the general flavone of various concentration (25,50,100 μM), H is measured with MTT measuring method2O2After (200 μM) are handled 4 hours
The survival rate of cell.
Fig. 4 is honeysuckle general flavone influence (mean ± SD, n=10) preclinical on rat in water maze laboratory
Note: compared with sham-operation group, * * p < 0.01；Compared with model group, ##p < 0.01.A, B, C respectively represent example
Three kinds of various doses of middle honeysuckle general flavone.
Fig. 5 for general flavone in honeysuckle to rat in water maze laboratory target quadrant to the time influence (mean ±
SD, n=10).
Note: compared with sham-operation group, * * p < 0.01；Compared with model group,##P < 0.01.A, B, C respectively represent example 5
Three kinds of various doses of middle honeysuckle general flavone
Fig. 6 be honeysuckle in general flavone to rat in water maze laboratory pass through position of platform number influence (mean ± SD,
N=10).
Note: compared with sham-operation group, * * p < 0.01；Compared with model group, ##p < 0.01.A, B, C respectively represent example 5
Three kinds of various doses of middle honeysuckle general flavone.
Specific embodiment
The present invention is further described in conjunction with specific embodiments.
The preparation of 1 honeysuckle general flavone of embodiment
Extracting honeysuckle dries medicinal material 1kg, heats (100 DEG C) extractions with water and obtains Aqueous extracts, and is concentrated, and it is appropriate to be added
Ethyl alcohol (ethanol content 70%) repeated multiple times sedimentation, remove its insoluble matter, be finally dried to obtain medicinal extract；By medicinal extract 1L
8 volumes are first eluted with water in upper CHP20SS resin after water dissolution, then with volume fraction are 70% ethyl alcohol water elution, until flowing out
Eluent be it is colourless.All 70% ethanol water eluents are collected, recycling ethyl alcohol is concentrated under reduced pressure, spray drying obtains xeraphium
End obtains 82.3g general flavone, utilizes FeCl3Colour developing is positive, and HCl-Mg reaction is positive, and determines that extract obtained is total
Flavones.It is adopted using visible spectrophotometry (one V A of annex of Chinese Pharmacopoeia) with Hyperoside (purity 98%) for reference substance
It is extracted with ethyl acetate, sodium carbonate, which is added, makes strong acid weak base combine the most chlorogenic acid ingredients of removing, effectively eliminates dry
The influence of ingredient is disturbed, and uses sodium nitrite-aluminum nitrate-sodium hydroxide colorimetric method, is reacted according to flavone compound with aluminium salt
Red complex, Detection wavelength 510nm are generated, testing result is shown in gained fraction, general flavone content 84.7%.
The preparation of 2 honeysuckle general flavone of embodiment
Extracting honeysuckle dries medicinal material 10kg, obtains Aqueous extracts with water heating extracting, and the pH value that hydrochloric acid adjusts Aqueous extracts is added
It after 5.00, then is concentrated, ethyl alcohol appropriate (ethanol content 70%) is added and is repeated several times sedimentation, removes its insoluble matter
Matter is finally dried to obtain medicinal extract；Upper CHP20P resin, is first eluted with water 4 volumes, then use body after medicinal extract is dissolved with 10L water
Fraction be 10%~20% 5 volumes of ethanol water system elutions, then with 75% 10 volumes of ethyl alcohol water elution, finally
Eluent with 90% ethanol water solvent system elutions to outflow is colourless.All 75% ethanol water eluents are collected, are depressurized
Concentration and recovery ethyl alcohol, freeze-drying obtain dried powder, obtain 0.85kg general flavone, utilize FeCl3Colour developing is positive, and HCl-
Mg reaction is positive, and determines extract obtained for general flavone.Using visible spectrophotometry (one V A of annex of Chinese Pharmacopoeia),
It with Hyperoside (purity 98%) for reference substance, adopts and is extracted with ethyl acetate, sodium carbonate, which is added, makes strong acid weak base combine removing exhausted
Most of chlorogenic acid ingredient effectively eliminates the influence of interference component, and uses sodium nitrite-aluminum nitrate-sodium hydroxide ratio
Color method reacts with aluminium salt according to flavone compound and generates red complex, Detection wavelength 510nm, testing result display gained
In fraction, general flavone content 85.4%.
The preparation of 3 honeysuckle general flavone of embodiment
Extracting honeysuckle dries medicinal material 30kg, obtains Aqueous extracts with water heating extracting, and the pH value that hydrochloric acid adjusts Aqueous extracts is added
After 4.00, ethyl alcohol appropriate (ethanol content 70%) is added and is repeated several times sedimentation, removes its insoluble matter, finally dry
To medicinal extract；Upper CHP20Y resin after medicinal extract is dissolved with 25L water, first elutes 5 volumes with 20% ethanol water elution system, then
It is 60%~80% ethyl alcohol water elution with volume fraction, until the eluent of outflow is colourless.Collect all 60%~80% second
Alcohol water elution, is concentrated under reduced pressure recycling ethyl alcohol, and vacuum drying obtains dried powder, obtains 2.68kg general flavone, utilize FeCl3It is aobvious
Color is positive, and HCl-Mg reaction is positive, and determines extract obtained for general flavone.Utilize visible spectrophotometry (middle traditional Chinese medicines
One V A of annex of allusion quotation), it with Hyperoside (purity 98%) for reference substance, adopts and is extracted with ethyl acetate, sodium carbonate, which is added, makes strong acid
Weak base, which combines, removes most chlorogenic acid ingredients, effectively eliminates the influence of interference component, and use sodium nitrite-nitric acid
Aluminium-sodium hydroxide colorimetric method reacts with aluminium salt according to flavone compound and generates red complex, Detection wavelength 510nm, detection
As the result is shown in gained fraction, general flavone content 86.5%.
The preparation of 4 honeysuckle general flavone of embodiment
Extracting honeysuckle dries medicinal material 50kg, obtains Aqueous extracts with water heating extracting, and be concentrated, ethyl alcohol appropriate is added
(ethanol content 70%) is repeated several times sedimentation, removes its insoluble matter, is finally dried to obtain medicinal extract；Medicinal extract is water-soluble with 60L
Upper CHP20Y resin is added 0.1% to improve the separating effect of CHP20Y resin in ethanol water eluent in advance after solution
Formic acid elutes 7 volumes with 10%, 20% ethanol water (containing 0.1% formic acid) elution system first, then is with volume fraction
80% ethanol water (containing 0.1% formic acid) elution, until the eluent of outflow is colourless.Collect all 80% ethyl alcohol water elutions
Liquid (contains 0.1% formic acid), and recycling ethyl alcohol is concentrated under reduced pressure, and normal heating is dried to obtain the higher total flavonoids extract of purity
4.12kg utilizes FeCl3Colour developing is positive, and HCl-Mg reaction is positive, and determines extract obtained for general flavone.Using visible
Spectrophotometry (one V A of annex of Chinese Pharmacopoeia) is adopted and is extracted with ethyl acetate, add with rutin (purity 98%) for reference substance
Entering sodium carbonate makes strong acid weak base combine the most chlorogenic acid ingredients of removing, effectively eliminates the influence of interference component, and adopt
With sodium nitrite-aluminum nitrate-sodium hydroxide colorimetric method, is reacted according to flavone compound with aluminium salt and generate red complex, inspection
Wavelength 510nm is surveyed, testing result is shown in gained fraction, general flavone content 86.1%.
The preparation of 5 honeysuckle general flavone of embodiment
Extracting honeysuckle dries medicinal material 100kg, obtains Aqueous extracts with water heating extracting, and the pH value that hydrochloric acid adjusts Aqueous extracts is added
After 3.50, low temperature concentration is depressurized, ethyl alcohol appropriate (ethanol content 70%) is added and is repeated several times sedimentation, removes it not
Molten substance, is finally dried to obtain medicinal extract；Upper CHP20Y resin after medicinal extract is dissolved with 90L water, in order to improve CHP20Y resin
0.1% phosphoric acid is added in separating effect in ethanol water eluent in advance, and it is a (containing 0.1% phosphoric acid) that 10 are eluted with water first
Volume, then eluted with volume fraction for 65% ethanol water (containing 0.1% phosphoric acid), until the eluent of outflow is colourless.It collects
All 65% ethanol water eluents (containing 0.1% phosphoric acid), are concentrated under reduced pressure recycling ethyl alcohol, obtain the total of high-purity after constant pressure and dry
Flavonoid extract 7.45kg.Utilize FeCl3Colour developing is positive, and HCl-Mg reaction is positive, and determines extract obtained for general flavone.
Using visible spectrophotometry (one V A of annex of Chinese Pharmacopoeia), with rutin (purity 98%) for reference substance, using ethyl acetate
Extraction, sodium carbonate, which is added, makes strong acid weak base combine the most chlorogenic acid ingredients of removing, effectively eliminates the shadow of interference component
It rings, and uses sodium nitrite-aluminum nitrate-sodium hydroxide colorimetric method, react the red network of generation with aluminium salt according to flavone compound
Object, Detection wavelength 510nm are closed, testing result is shown in gained fraction, general flavone content 85.9%.
Embodiment 6
The honeysuckle general flavone that different instances are prepared anti-A amyloid beta aggregation activity evaluation in vitro
1, the pretreatment of A β
The A β in -80 DEG C of refrigerators will be pre-saved to take out, be dissolved in HFIP with the concentration of 1mg/mL.It stands at room temperature
For 30min after its secondary structure completely removes, vacuum freeze drying removes HFIP, is placed in -20 DEG C of refrigerators and saves.
2, prepared by phosphate buffer (PBS, pH=7.4,0.1M)
The preparation (PBS, buffer) of buffer: pH=7.4, concentration 0.1mol/mL.Referring to People's Republic of China's medicine
Allusion quotation second annex of version in 2010, takes potassium dihydrogen phosphate 1.36g, adds 0.1mol/L sodium hydroxide solution 79mL, be diluted with water to
200mL, with Accurate pH acidometer be corrected to get.
3, the preparation of A beta response test solution
Pretreated A β freeze-dried powder is taken, is tried with the A β that the 0.1mol/mL PBS of pH=7.4 is configured to 50 μm of ol/L
Liquid, -20 DEG C freeze it is spare.
4, color developing agent configures
Precision weighs Th-T powder, with the 0.1mol/mL PBS of pH=7.4, prepares the Th-T test solution for becoming 10 μm of ol/L,
Matching while using.
5, the preparation of compound solution
With the 0.1mol/mL PBS and DMSO of pH=7.4, it is 10 that concentration, which is respectively configured, in VE and sample to be tested-3, 10-4,
10-5, 10-6With 10-7The final volume score of mol/L, DMSO are not more than 0.001.
6, measuring method
Group setting:
(1) blank control group: 60 μ L, A β of PBS, 10 10 μ L of μ L, PBS+DMSO
(2) sample inhibition group: 60 μ L, A β of PBS 10 μ L, 10 μ L of drug
(3) blank background group: 60 10 10 μ L of μ L, PBS+DMSO of μ L, PBS of PBS
(4) sample copy bottom group: PBS 60 μ L, PBS 10 μ L, 10 μ L of drug
It is separately added into above-mentioned each group solution in 96 orifice plates, is placed in constant-temperature incubation case and is incubated for for 24 hours, then into each group for 37 DEG C
80 μ L Th-T solution are added, 37 DEG C of incubation 5min in constant-temperature incubation case are placed in, with microplate reader in (λ ex=450nm；λ em=
485nm) place measures each group fluorescent value.Every group is repeated 3 times.
7, interpretation of result:
Fluorescent value is as control at the 485nm for using Th-T to measure after being individually incubated for using A β: in order to avoid compound itself has
Interference of some fluorescence to result uses the fluorescent value of Th-T measurement as background to deduct independent test-compound.Inhibiting rate
Calculation formula is as follows:
8, IC is calculated50Value
Data are analyzed by SPSS16.0, calculate IC50Value.
9, Activity Results are shown in Fig. 1.
Embodiment 7
The protective effect for the SY5Y cell that general flavone mediates hydrogen peroxide in honeysuckle is tested
Cell: SH-SY5Y
Model: hydrogen peroxide induced oxidation damage
Detailed process:
1. bed board: choosing the good clean SY5Y cell of growth conditions, be taped against 96 orifice plates and cultivated, 12h is spread before sieving medicine
Plate, optional 1.2*104/hole of cell density.(bed board is uniform, appropriate density)
2. making up a prescription: after 12h, whether observation cell is adherent, and whether growth conditions are good, such as qualified, can carry out dosing.
Drug concentration has been calculated in advance according to concentration group number and every group of secondary orifices number, and (Stock concentrations are culture volume
50mM).From high concentration → low concentration dilution when making up a prescription.Pay attention to mixing.
3. dosing: culture plate being taken out from incubator, has marked group, knockout plate in plate lid.By the medicine prepared by low dense
Degree → high concentration sequentially adds in hole (pipette tips resist side wall when dosing, slowly squeeze into, and can not impact cell).
4. adding hydrogen peroxide modeling: calculating hydrogen peroxide concentration in advance according to cell density.Add hydrogen peroxide (dioxygen after dosing 1h
Water easily decomposes, ready-to-use).Hydrogen peroxide is made into 8.8mM stock solution first, then is diluted to required concentration with culture medium, is paid attention to
It mixes (fast operating).
Dosing group and model group (being slowly added to) is added in the hydrogen peroxide prepared.Control group plus isometric culture medium
Make blank control.
5. adding MTT: adding after hydrogen peroxide in 1-2, when observing and nursing group cellular damage rate is up to 60% or so, add MTT.
6. surveying OD value: adding after MTT in 2-4h, drafting board.
7. handling data: according to the OD value measured, calculating cell survival rate, experimental result is as shown in Figures 2 and 3.
The water maze laboratory of 8 Senile Dementia Model Rats of embodiment
SD rat is taken, male, 250~300g of weight establishes rat Senlie dementia model, and animal is randomly divided into 6 groups:
Sham-operation group: animal not modeling, stomach-filling is to physiological saline, once a day, successive administration 30 days；
Model group: after animal model, stomach-filling is to physiological saline, once a day, successive administration 30 days；
Positive controls: after animal model, donepezil 1.5mg/kg is given in stomach-filling, once a day, successive administration 30 days；
Honeysuckle general flavone low dose group: after animal model, honeysuckle general flavone 100mg/kg is given in stomach-filling, once a day,
Successive administration 30 days；
Honeysuckle general flavone middle dose group: after animal model, honeysuckle general flavone 200mg/kg is given in stomach-filling, once a day,
Successive administration 30 days；
Honeysuckle general flavone high dose group: after animal model, honeysuckle general flavone 400mg/kg is given in stomach-filling, once a day,
Successive administration 30 days.
Using the cognitive function of water maze laboratory observation rat.Water maze instrument is by a round pool and a moveable circle
Shape platform composition, pool diameter 100cm, depth of water 40cm, water temperature are controlled at 23 ± 1 DEG C, and platform diameter 15cm, the water surface is higher by platform
2cm.When experiment, platform is placed in the centre of first quartile, and automatic camera system is arranged above pond and connects computer, tracks
The swimming track for recording rat, calculates the distance and find the time required for platform that rat swims across in pond automatically.Just
Before formula experiment, animal is placed on platform and adapts it to 10s, then into the water by rat, observes the movement feelings in 90s
Condition.Record rat finds the time required for platform, if not finding platform in animal 90s, it is gently placed on platform simultaneously
Continue 15s, then puts back in cage.Daily animal training three times, for three days on end, then starts formally to test.Observation rat is put into water certainly
Chi Zhongzhi finds the time required for platform, and more than 90s, then the time is calculated as 90s, once a day, continuous 5 days.After experiment,
Each animal is taken a day off, then carries out memory experiment.Platform is withdrawn from, rat is put into pond from third quadrant, observes and records
Each rat is in the time where first quartile and by the number of platform position in 120s.
The results show that in experiment in 5 days, the time for finding platform obviously increases model group rats compared with sham-operation group
(incubation period), across first quartile number and first quartile to time be decreased obviously.Compared with model group, honeysuckle is total
Flavones 200mg/kg, 400mg/kg dosage group can obviously reduce the incubation period of rat, increases the number by first quartile and prolongs
Grow first quartile to time, difference has conspicuousness (P < 0.01).It is similar how its effect is organized together to more piperazines.Experimental result
As shown in Fig. 4,5 and 6.
Claims (10)
1. honeysuckle general flavone, which is characterized in that be prepared via a method which: by the side of honeysuckle medicinal part water extract-alcohol precipitation
Method extracts, and is suspended after being concentrated under reduced pressure to give crude extract with water, and by CHP20 column chromatography column, it is always yellow finally to obtain honeysuckle
Ketone.
2. the preparation method of honeysuckle general flavone as described in claim 1, characterized in that
(1) traditional Chinese medicine honeysuckle, with water heating and refluxing extraction；
(2) it is concentrated under reduced pressure, industrial alcohol is added in concentrate, filters after standing, recycles ethyl alcohol, obtains medicinal extract；
(3) after adding water to be suspended gained medicinal extract, upper HP20 resin, first with 10%~20% ethyl alcohol water elution, then with 60%~
80% ethyl alcohol water elution collects 60%~80% ethanol water eluent, ethyl alcohol is recovered under reduced pressure, is drying to obtain.
3. preparation method as claimed in claim 2, which is characterized in that
In step (1), first with the water soaked overnight of 2-3 times of volume, then water heating extracting 2-3 times with 5-8 times of volume.
4. preparation method as claimed in claim 2, which is characterized in that
In step (2), the pH value that extracting solution can be adjusted before reduced pressure is 3.00~5.00.
5. preparation method as claimed in claim 2, which is characterized in that
In step (2), aqueous extract is concentrated into 1:1, ethyl alcohol, which is added, makes its alcohol content 70-80%.
6. preparation method as claimed in claim 2, which is characterized in that
In step (3), the HP20 is one or more of CHP20Y, CHP20P, CHP20SS, the ethanol elution
Acid can be added in liquid.
7. honeysuckle general flavone as described in claim 1, which is characterized in that content assaying method are as follows: use vis spectroscopy light
Degree method is adopted and is extracted with ethyl acetate using 98% Hyperoside of purity as reference substance, and the shadow of sodium carbonate exclusive PCR ingredient is added
It rings, and uses sodium nitrite-aluminum nitrate-sodium hydroxide colorimetric method, react the red network of generation with aluminium salt according to flavone compound
Close object, Detection wavelength 510nm.
8. a kind of Pharmaceutical composition includes honeysuckle general flavone described in claim 1 and pharmaceutically acceptable carrier or tax
Shape agent.
9. honeysuckle general flavone described in claim 1 or Pharmaceutical composition according to any one of claims 8 inhibit aβ protein in preparation
Aggregation drug in application.
10. honeysuckle general flavone described in claim 1 or Pharmaceutical composition according to any one of claims 8 treat A Erzi in preparation
Application in the silent disease medicament in sea.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113288927A (en) * | 2021-05-26 | 2021-08-24 | 邵阳学院 | Process for extracting total flavonoids from honeysuckle |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2010106001A (en) * | 2008-10-31 | 2010-05-13 | Theravalues Corp | Ppar activator |
| CN103823034A (en) * | 2014-03-06 | 2014-05-28 | 山东省食品药品检验所 | Honeysuckle contrasting extract and preparation method thereof |
| CN108743599A (en) * | 2018-05-03 | 2018-11-06 | 东北师范大学 | A kind of application of compound in terms of reducing albumen and building up treatment senile dementia |
-
2018
- 2018-12-12 CN CN201811514987.3A patent/CN109464488B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2010106001A (en) * | 2008-10-31 | 2010-05-13 | Theravalues Corp | Ppar activator |
| CN103823034A (en) * | 2014-03-06 | 2014-05-28 | 山东省食品药品检验所 | Honeysuckle contrasting extract and preparation method thereof |
| CN108743599A (en) * | 2018-05-03 | 2018-11-06 | 东北师范大学 | A kind of application of compound in terms of reducing albumen and building up treatment senile dementia |
Non-Patent Citations (5)
| Title |
|---|
| PEIPEI WANG ET AL: "A glucan isolated from flowers of Lonicera japonica Thunb. inhibits aggregation and neurotoxicity of Aβ42", 《CARBOHYDRATE POLYMERS》 * |
| 侯燕等: "基于β淀粉样蛋白为靶标的中医药防治阿尔茨海默病实验研究 ", 《中国中医基础医学杂志》 * |
| 章晓骅: "金银花活性成份提取及颗粒剂制剂工艺研究", 《中国优秀博硕士学位论文全文数据库（硕士）工程科技I辑》 * |
| 胡海山等: "均匀试验设计优化金银花总黄酮的提取 ", 《现代测量与实验室管理》 * |
| 陈欣等: "金银花水溶性化学成分研究", 《中华中医药学会中药化学分会第五届学术年会》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113288927A (en) * | 2021-05-26 | 2021-08-24 | 邵阳学院 | Process for extracting total flavonoids from honeysuckle |
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