CN109498596A - A kind of composition and its preparation method and application for treating senile dementia - Google Patents
A kind of composition and its preparation method and application for treating senile dementia Download PDFInfo
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Abstract
The present invention provides a kind of compositions and its preparation method and application for treating senile dementia, belong to biomedicine technical field.Composition provided by the invention includes the component of following mass parts: 0.5~1 part of Co-Q10,0.05~0.2 part of polysorbate, 0.3~0.8 part of MCT Oil, 3~8 parts of gelatin, 3~8 parts of glycerol and 3~8 parts of sorbierites.Composition provided by the invention can effectively penetrate blood-brain barrier and enter brain deep, arrive at target neuron under the packing specification of≤50nm；Efficient activation neuron linear plastochondria, improves the vigor of mitochondrial cytochrome C, and enhancing ATP synthesis provides energy to neuron；Intracerebral formaldehyde is reduced, formaldehyde is reduced and the promotion formation of senile plaque is acted on, achieve the purpose that scavenger-cell gap blocks, restore the flowing of intercellular fluid；Nontoxic to the human body, the memory of Model of Dementia mouse can be improved in the ability with strengthen immunity.
Description
Technical field
The invention belongs to biomedicine technical fields, and in particular to a kind of for treating the composition and its system of senile dementia
Preparation Method and application.
Background technique
Senile dementia (Alzheimer ' s disease, AD) is a kind of nervous system of the progress sexual development of onset concealment
Degenerative disease.Clinically with memory disorders, aphasia, appraxia, agnosia, the damage of visual space technical ability, execution dysfunction and people
The performance of the generalized dementias such as lattice and behavior change is characterized, and the cause of disease is unknown so far.The whole world spends a few $ 100 billion expenses every year
AD drug is developed, fourth phase clinic is not all entered into.Dementia, which faces, to past medical help, and can not reverse the awkward situation of amnesia.At present
Clinical dull-witted medication such as Memantine, donepezil, curcumin etc. has significant curative effect at dull-witted initial stage, but after 5 years all gradually
Lose clinical efficacy.AD brings heavy economy and society to bear to each country, family.
Clinical research thinks that it is to induce heredity that A β extracellular deposition, which forms senile plaque and oligomerization generation toxicity intracellular,
The affirmative that family AD occurs, the crucial cause of disease.But the whole world is clinical most authoritative to declare for Britain's periodical " lancet " 2016
Cloth: current existing exploitation AD drug all fails.Either it is directed to generation, the aggregation, the antibody of elimination, small molecule chemical combination of A β
Object, in conjunction with the polypeptide of A β, all do not reach the curative effect of clinical expectation.These bitterness failures, high economic cost, force us
Considering ----what reason results in the failure of AD drug development.Existing viewpoint thinks, all 95% antibody, synthesis it is more
The main reason for peptide, macromolecular are all difficult to through blood-brain barrier, this is the AD drug failure of overwhelming majority exploitation.But also have
Research shows that: the AD drug of some exploitations can penetrate blood-brain barrier, it is difficult to which understanding is that drug cannot still save brain deep
The death of neuron.And the reason of our current research discloses drug failure at present: drug diameter is excessive and can not penetrate
The space between cells that 50 nanometers of < reaches the neuron in brain deep by the flowing of intercellular fluid, does not have the guarantor to neuron
Shield effect.
Summary of the invention
The problem of in view of background technique, can effectively treat senile dementia the purpose of the present invention is to provide a kind of
Composition and its preparation method and application.
The present invention provides a kind of for treating the composition of senile dementia, the component including following mass parts: 0.5~1
Part Co-Q10,0.05~0.2 part of polysorbate, 0.3~0.8 part of MCT Oil, 3~8 parts of gelatin, 3~8
Part glycerol and 3~8 parts of sorbierites.
Preferably, the composition includes the component of following mass parts: 0.7~0.85 part of Co-Q10,0.08~0.12 part
Polysorbate, 0.4~0.6 part of MCT Oil, 4~6 parts of gelatin, 4~6 parts of glycerol and 4~6 parts of sorbierites.
Preferably, the composition is pulvis, particle diameter≤50nm of the pulvis.
The present invention provides the preparation methods of above-mentioned composition, include the following steps:
(1) by Co-Q10, polysorbate, MCT Oil, gelatin, glycerol, sorbierite and dehydrated alcohol
Mixing, obtains organic phase；
(2) organic phase is mixed with the water of 3~8 times of organic phase volumes, obtains mixed liquor；
(3) mixed liquor is heated to 60~80 DEG C, stirring is concentrated by evaporation to 0.3~0.8 times of organic phase volume, is obtained
Nanoemulsion；
(4) nanoemulsion is placed in 0~4 DEG C of environment and continues to stir, obtain a nanometer Co-Q10 suspension；
(5) by after the nanometer Co-Q10 suspension homogeneous, vacuum freeze drying, the combination for the senile dementia that obtains medical treatment
Object.
Preferably, the ratio between the quality of Co-Q10 and the volume of dehydrated alcohol are 0.5~1g:100mL in the step (1).
Preferably, the step (3) or (4) middle rate stirred are 1000~2000rpm.
Preferably, the time stirred in the step (4) is 1~2h.
Preferably, the temperature of homogeneous is 45~55 DEG C in the step (5), and the pressure of homogeneous is 200~1500bar.
Preferably, the number of the homogeneous is 2~4 times, and the pressure of the homogeneous gradually increases.
The composition that the present invention also provides above-mentioned for treating senile dementia is in preparation treatment medicine for senile dementia
Using.
The utility model has the advantages that the present invention provides a kind of for treating the composition of senile dementia, the composition includes as follows
The component of mass parts: 0.5~1 part of Co-Q10,0.05~0.2 part of polysorbate, 0.3~0.8 part of medium chain fatty acid three
Ester, 3~8 parts of gelatin, 3~8 parts of glycerol and 3~8 parts of sorbierites.The research of the invention finds that: the senile plaque blocking that A beta-aggregation is formed
The space between cells of adjacent neurons (Extracellular space, ECS), has blocked intercellular fluid (interstitial
Fluid, ISF) flowing from from the superficial layer of brain to brain deep.This means that the blocking in neuronal cell gap is that drug can not be with
Intercellular fluid enters the key reason in brain deep.Accordingly, the present invention is less than the drug of 50nm by exploitation diameter, with iuntercellular
The flowing of liquid reaches brain deep, and plays the dull-witted purpose for the treatment of, provides new thinking to treatment dementia.It is provided by the invention
Composition can effectively penetrate blood-brain barrier and enter brain deep, arrive at target neuron under the packing specification of≤50nm；Efficient activation
Neuron linear plastochondria, improves the vigor of mitochondrial cytochrome C, and enhancing ATP synthesis provides energy to neuron；Reduce intracerebral
Formaldehyde reduces formaldehyde and acts on the promotion formation of senile plaque, achievees the purpose that scavenger-cell gap blocks, restore intercellular fluid
Flowing；Nontoxic to the human body, the memory of Model of Dementia mouse can be improved in the ability with strengthen immunity.
Detailed description of the invention
Fig. 1 is schematic diagram of the composition provided by the invention when penetrating blood-brain barrier；
Fig. 2 is treatment schematic diagram of the composition provided by the invention in treatment senile dementia processes；
Fig. 3 is testing result in the embodiment of the present invention 3；
Fig. 4 is testing result in the embodiment of the present invention 4；
Fig. 5 is testing result in the embodiment of the present invention 5；
Fig. 6 is testing result in the embodiment of the present invention 6.
Specific embodiment
The present invention provides a kind of for treating the composition of senile dementia, the component including following mass parts: 0.5~1
Part Co-Q10,0.05~0.2 part of polysorbate, 0.3~0.8 part of MCT Oil, 3~8 parts of gelatin, 3~8
Part glycerol and 3~8 parts of sorbierites.
Composition provided by the invention includes Co-Q10, polysorbate, MCT Oil, gelatin, glycerol
And sorbierite.In the present invention, in parts by mass, the dosage of the Co-Q10 be 0.5~1 part, preferably 0.7~0.85 part,
More preferably 0.78 part；The dosage of the polysorbate be 0.05~0.2 part, preferably 0.08~0.12 part, more preferably
0.1 part；The dosage of the MCT Oil is 0.3~0.8 part, preferably 0.4~0.6 part, more preferably 0.5
Part；The dosage of the gelatin is 3~8 parts, preferably 4~6 parts, more preferably 5 parts；The dosage of the glycerol is 3~8 parts, excellent
It is selected as 4~6 parts, more preferably 5 parts；The dosage of the sorbierite is 3~8 parts, preferably 4~6 parts, more preferably 5 parts.At this
In invention, above-mentioned each component and consumption proportion can effectively improve Co-Q10 encapsulation ratio and drugloading rate (especially medicine rouge ratio=
In the case where 1:20, effect is more preferable).The present invention is not particularly limited the source of above-mentioned each component, and this field conventional commercial produces
Product.In an embodiment of the present invention, the purchasing channel correspondence of each component is as follows: Co-Q10 (CAS:303-98-0, Xi'an
The four seasons Biotechnology Co., Ltd), polysorbate -80 (CAS:9005-65-6, Hubei Guang Ao Biotechnology Co., Ltd),
MCT Oil (CAS:73398-61-5, Shanghai Shu Can Industrial Co., Ltd.), gelatin (CAS:9000-70-8, Zheng
State Qi Huadun chemical products Co., Ltd), glycerol (CAS:56-81-5, Shanghai Yi En chemical technology Co., Ltd), sorbierite
(CAS:142-77-8, U.S. haze chemistry brand are Mei Lan industry (Shanghai) Co., Ltd.), dehydrated alcohol (CAS:64-17-5, Shanghai
Putuo branch company, Fang Ye Chemical Co., Ltd.).
In the present invention, the dosage form of the composition is preferably pulvis.The particle diameter of the pulvis preferably≤50nm, more
It is preferred that≤40nm, more preferably 25~35nm.The research of the invention finds that diameter≤50nm packaging Co-Q10 can effectively be worn
Saturating blood-brain barrier enters brain deep, efficient to activate neuron linear plastochondria, generates ATP, improves the memory of dementia mice.
The present invention provides the preparation methods of above-mentioned composition, include the following steps:
(1) by Co-Q10, polysorbate, MCT Oil, gelatin, glycerol, sorbierite and dehydrated alcohol
Mixing, obtains organic phase；
(2) organic phase is mixed with the water of 3~8 times of organic phase volumes, obtains mixed liquor；
(3) mixed liquor is heated to 60~80 DEG C, stirring is concentrated by evaporation to 0.3~0.8 times of organic phase volume, is obtained
Nanoemulsion；
(4) nanoemulsion is placed in 0~4 DEG C of environment and continues to stir, obtain a nanometer Co-Q10 suspension；
(5) by after the nanometer Co-Q10 suspension homogeneous, vacuum freeze drying, the combination for the senile dementia that obtains medical treatment
Object.
The present invention is first by Co-Q10, polysorbate, MCT Oil, gelatin, glycerol, sorbierite and nothing
Water-ethanol mixing, obtains organic phase.In the present invention, the ratio between the quality of the Co-Q10 and the volume of dehydrated alcohol are preferably
0.5~1g:100mL, more preferably 0.7~0.85g:100mL, more preferably 0.78g:100mL.The present invention is to above-mentioned each component
Order by merging be not particularly limited.In the present invention, after the mixing it is preferable to use ultrasonic treatment, the ultrasonic treatment it is strong
Degree is preferably 150~250W, more preferably 200W.The time of the ultrasonic treatment is preferably 15~25min, more preferably
20min.It is fully dispersed in dehydrated alcohol that the ultrasonic treatment is more advantageous to each component.
After obtaining organic phase, the present invention mixes the organic phase with water, obtains mixed liquor.In the present invention, the water
Preferably distilled water, the dosage of the water are preferably 3~8 times of organic phase volume, more preferably 5 times.Present invention preferably employs
Organic phase is slowly injected into water by syringe.In injection process, the present invention is preferably stirred.The rate of the stirring is preferred
For 1000~2000rpm, more preferably 1500rpm.
After obtaining mixed liquor, the present invention heats the mixed liquor, and stirring is concentrated by evaporation, and obtains nanoemulsion.In this hair
In bright, the temperature of the heating is preferably 60~80 DEG C, more preferably 65~75 DEG C, more preferably 70 DEG C.The speed of the stirring
Rate is preferably 1000~2000rpm, more preferably 1500rpm.The heating and stirring can promote the evaporation of solvent in mixed liquor,
Realize the concentration of mixed liquor.In the present invention, the degree of the concentration is preferably concentrated into 0.3~0.8 times of organic phase volume, more excellent
Choosing is concentrated into 0.5 times of organic phase volume.
Nanoemulsion is obtained after concentration.The nanoemulsion is placed in by the present invention to be continued to stir in low temperature environment, is received
Rice Co-Q10 suspension.In the present invention, the low temperature stirring power wraps up Co-Q10 rapidly, improves drugloading rate.At this
In invention, the low temperature is preferably 0~4 DEG C, and more preferably 0~2 DEG C.The low temperature environment is preferably ice-water bath.The continuation
The rate of stirring is preferably 1000~2000rpm, more preferably 1500rpm.The time for continuing stirring is preferably 1~2h,
More preferably 1.5h.
The suspension of nanometer Co-Q10 is obtained after stirring.The present invention is by the nanometer Co-Q10 suspension homogeneous.At this
In invention, the temperature of the homogeneous is preferably 45~55 DEG C, and more preferably 50 DEG C.The pressure of the homogeneous is preferably 200~
1500bar.In the present invention, the number of the homogeneous is preferably 2~4 times, and more preferably 3 times.The pressure of the homogeneous is gradually
Increase, when homogenization cycles be 3 times when, the pressure of the homogeneous successively be preferably 250~350bar, 700~900bar, 1100~
1300bar, more preferably 300bar, 800bar, 1200bar.
For the present invention by after the nanometer Co-Q10 suspension homogeneous, vacuum freeze drying obtains white powder after dry.
The present invention is 30~40nm using the diameter of electron cryo-microscopy detection white powder.After obtaining white powder, the present invention preferably will be white
Color powder is placed in 4 DEG C of closed environments and saves.The white powder is the composition for the treatment of senile dementia of the present invention.Such as
Shown in Fig. 1, the diameter of white powder is less than space between cells, can penetrate blood-brain barrier and enter brain deep.
The present invention also provides application of the above-mentioned composition in preparation treatment medicine for senile dementia.In the present invention, institute
Stating drug is preferably capsule；The capsule using oral into prescription formula, preferably one day one.In the present invention, every
Capsule preferably comprises 0 microgram of nanometer DPN diphosphopyridine nucleotide.In the present invention, in the capsule finished product nanometer Co-Q10 mass content
Preferably 5~20%, more preferably 10%.
In the present invention, the treatment principle of the application is as shown in Figure 2.Wherein, molecular mechanism occurs for AD are as follows: familial is silly
Excessive A β, the A β of slow-witted generation is deposited on the space between cells (ECS) between two neurons and forms senile plaque (SP), and then blocks cell
Between liquid (ISF) flowing.Energy oligomerization enters cell mitochondrial to A β again, in conjunction with formaldehyde dehydrogenase FDH, and inhibits FDH vigor,
Cause formaldehyde that cannot degrade and accumulate.Excessive formaldehyde collaboration A β induces intracellular Ca2+ and increases, and generates excessive active oxygen and induces
Neuronal death.Composition provided by the invention has the function for the treatment of AD: on the one hand ,≤50nm (especially 30nm) packaging
Co-Q10 can reduce intracerebral formaldehyde, reduce formaldehyde and act on the promotion formation of senile plaque, reach scavenger-cell gap blocking mesh
, restore the flowing of intercellular fluid；On the other hand, the Co-Q10 of≤50nm (especially 30nm) packaging can be with space between cells
Intercellular fluid flow to the deep of brain, provide ATP energy in apoptosis or dead neuronal, mitochondrial function be provided, is drawn
The vigor of neuron is rescued, to gradually restore brain memory ability.
Experiment shows that composition provided by the invention can effectively penetrate blood-brain barrier and enter brain deep, arrives at target neuron；
Efficient activation neuron linear plastochondria, improves the vigor of mitochondrial cytochrome C, and enhancing ATP synthesis provides energy to neuron；
Intracerebral formaldehyde is reduced, formaldehyde is reduced and the promotion formation of senile plaque is acted on, achieve the purpose that scavenger-cell gap blocks, restore thin
The flowing of tissue fluid；Nontoxic to the human body, the memory of Model of Dementia mouse can be improved in the ability with strengthen immunity.
Below with reference to embodiment to a kind of composition and preparation method thereof for treating senile dementia provided by the invention
It is described in detail with application, but they cannot be interpreted as limiting the scope of the present invention.
Embodiment 1
Prepare the Co-Q10 of nano-packaging
Primary raw material: Co-Q10 (CAS:303-98-0, the Xi'an four seasons Biotechnology Co., Ltd), polysorbate -80
(CAS:9005-65-6, Hubei Guang Ao Biotechnology Co., Ltd), MCT Oil (CAS:73398-61-5, on
Hai Shucan Industrial Co., Ltd.), gelatin (CAS:9000-70-8, Zhengzhou Qi Huadun chemical products Co., Ltd), glycerol (CAS:
56-81-5, Shanghai Yi En chemical technology Co., Ltd), (CAS:142-77-8, U.S. haze chemistry brand are U.S. haze industry to sorbierite
(Shanghai) Co., Ltd.), dehydrated alcohol (CAS:64-17-5, Shanghai Putuo branch company, Fang Ye Chemical Co., Ltd.), purified water
(laboratory deionized water).
Preparation method: the Co-Q10 of nano-packaging is prepared using emulsification-low temperature-high-pressure process of original creation
1. accurately claim 0.78g Co-Q10,0.1g polysorbate -80,0.5g MCT Oil, 5g gelatin,
5g glycerol, 5g sorbierite；
2. being dissolved in obtained each component is weighed in 100mL dehydrated alcohol, 200W ultrasound 10 minutes, is constituted organic twice
Phase；
3. being slowly injected to organic phase with syringe in the distilled water of 200mL stirring (1500r/min), 70 DEG C of constant temperature are stirred
It mixes, evaporate solvent and is concentrated into 50mL, obtain translucent nanoemulsion；
4. nanoemulsion is quickly adding into the ice-water bath container of 50mL (0-2) DEG C, continue to stir 1.5h to get to receiving
Rice Co-Q10 suspension；
5. by obtained nanometer Co-Q10 suspension (thick cream) high-pressure homogeneous circulation 3 times, homogenization pressure is respectively
300bar, 800bar and 1200bar.Homogenizing temperature is 50 DEG C, then carries out vacuum freeze drying.White powder after drying is
It is the Co-Q10 of nano-packaging.Chilled Electronic Speculum detection is directly 30-40 nanometers.Closed 40C is saved.
Embodiment 2
Nano-packaging Co-Q10 penetrates blood-brain barrier research
Detection method: high performance liquid chromatography (HPLC)
Detection process:
Instrument and reagent: HP1100 high performance liquid chromatograph (HP1100, Agilent Technologies Co.Ltd,
USA)；Nitrogen flushing instrument (KL512J, Beijing health science and technology limited Company).(BBI company, purity are greater than Co-Q10 standard items
99.9%).The preparation of Co-Q10 standard solution: accurate respectively to claim Co-Q10 0.0250g in 25mL volumetric flask, Co-Q10
With ethyl alcohol dissolution and constant volume is to scale, shakes up spare, is made into the standard reserving solution of 1g/L.Standard curve is pressed with mobile phase before measurement
It is diluted to and uses concentration.
Chromatographic condition: chromatographic column selects PhenomenexC18 column (5 μm, 250mm × 4.6mm i.d.)；With methanol/ethanol
(volume ratio 20: 80) is mobile phase；Detection wavelength changeover program: 0~8min is 292nm, and 8~16min is 275nm；Flow velocity
1L/min；20 μ L of sample volume；25 DEG C of column temperature.
The drafting of standard curve: drawing appropriate vitamin E and Co-Q10 standard reserving solution, be diluted to 0 with mobile phase,
2.0, the standard series of 5.0,10.0,20.0,50.0mg/L takes 20 μ L sample introductions to measure, with standard solution mass concentration for horizontal seat
Mark, peak area are that ordinate draws standard curve.
Sample treatment: it is light-exposed degradable because containing isopentene group in Co-Q10 structure, therefore sample pretreatment is being protected from light
Under the conditions of carry out.It takes 0.2mL blood sample in centrifuge tube, 0.2mL ethanol precipitation albumen is added, vortex oscillation mixes 1min.It is added
1mL n-hexane, vortex oscillation mix 2min.It takes 800 μ L of supernatant in centrifuge tube, is dried with nitrogen.Mobile phase (methanol/ethanol
Volume ratio 20: 80) dissolved residue, 12000r/min are centrifuged 2min, and 50 μ L sample introduction of supernatant is taken to measure.
Testing location: Institute of Biophysics, Academia Sinica；
Testing result is as shown in figure 3, the brain Co-Q10 content of mouse increases about 3 times after stomach-filling in 15 days.
Embodiment 3
Effect of the nano-packaging Co-Q10 to ATP
Detection method: (96T, mouse atriphos (ATP) ELISA kit, Shanghai innermost thoughts and feelings are raw for mouse ATP kit
Object Science and Technology Ltd.)
Program: Mice brain tissues sample is detected, after cutting sample, weighs weight.A certain amount of PBS, PH7.4 is added.With
Liquid nitrogen quick freeze saves backup.Sample still maintains 2-8 DEG C of temperature after melting.It is added a certain amount of PBS (PH7.4), or
Histone extraction agent, by hand or homogenizer is by sample homogenization.It is centrifuged 20 minutes or so (2000-3000 revs/min).It is young
It is thin to collect supernatant.A to be detected after packing, remaining freezing is spare, shows that program is detected according to kit.
Testing result is as shown in figure 4, that improves that brain ATP content less packs after illustrating acute injection rapidly reaches 3 times
As many as.
Embodiment 4
Effect of the nano-packaging Co-Q10 to intracerebral formaldehyde
Detection method: high performance liquid chromatography (HPLC)
Detect program:
Material: Mouse Whole Brain, acetonitrile (liquid chromatographic grade is pure, Fisher Scientific, the U.S.), 2,4- dinitrobenzenes
Hydrazine (analyzes pure, Beijing Inst. of Chemical Reagent), and formaldehyde (analyzes pure, Sigma-Aldrich), trichloroacetic acid (point
Analyse pure, Xilong Chemical Co., Ltd), methylene chloride (analyzes pure, Beijing Chemical Plant), and sodium chloride (analyzes pure, western Gansu Province chemical industry
Limited liability company), acetic acid (analyzes pure, Xilong Chemical Co., Ltd).
Reagent: 2,4-dinitrophenylhydrazine acetonitrile solution (1.0g/L): 2,4-dinitrophenylhydrazine 100mg is weighed, acetonitrile is dissolved in
It is settled to 100ml.10% solution of trichloroacetic acid: weighing trichloroacetic acid 10g, is settled to 100ml with ultrapure water.
Sample preparation: the preparation of brain sample: taking each 1g of different parts brain tissue, and 10% solution of trichloroacetic acid is added
(10ml), tissue homogenate, centrifugation (13000r/min, 4 DEG C, 30min).Take 0.4ml supernatant, 0.1ml 2,4 dinitrophenyl hydrazine
(1g/L), 0.5ml acetonitrile.After mixing, 60 DEG C of heat preservation 30min. are centrifuged (13000r/min, 4 DEG C, 10min), and supernatant is taken to be used for
HPLC analysis.Retain extraction step (control): the methylene chloride of 2 times of volumes, vortex vortex oscillation 1min, centrifugation is added
(3000r/min, room temperature, 10min), takes the extract liquor of lower layer, is placed in tool plug glass tube.80 DEG C of 1~2h of water-bath, liquid to be extracted
All be evaporated after, be cooled to room temperature be added 1ml acetonitrile, vortex vortex oscillation 1min, centrifugation (3000r/min, room temperature,
10min), supernatant is taken to analyze for HPLC.High-efficient liquid phase chromatogram condition: LC-20A high performance liquid chromatograph UV-HPLC, SPD-
M20A diode array detector (Shimadzu Corporation, Japan).Chromatographic column: LiChrospher 100RP-18 (250mm x4.6mm
X5 μm) (Merck company, Germany)；Mobile phase: Yi Jing ︰ ultrapure water=65 ︰ 35 (v/v), ultrasonic degassing；Flow velocity: 0.8ml/min；
Sample volume: 20 μ l；Detection wavelength: 355nm；Column temperature: 35 DEG C of
Testing location: Institute of Biophysics, Academia Sinica；
Testing result is as shown in Figure 5: nano-packaging Co-Q10 can reduce rapidly mouse brain first in acute stomach-filling mouse
The content of aldehyde.
Embodiment 5
The raising effect that nano-packaging Co-Q10 remembers APP mouse
Detection method: Morris water maze
Morris water maze laboratory system is made of round pool, image automatic collection and processing system.Image is adopted automatically
Collection and processing system are mainly made of video camera, computer, picture monitor, and animal starts monitoring device after entering water, and record is dynamic
Object motion profile, experiment, which finishes, automatically analyzes reporting of relevant parameters.Morris water maze round pool is a circle ABS engineering plastic
Expect bucket, diameter 100cm, a height of 50-60cm, the pond depth of water is 30-40cm, and a circular platform diameter is 9cm, conceal in
Under the water surface of 2cm.The inside in pond must not have any label, and suitable fresh milk or milk powder are added in water, make pond at
For opaque milky.East, south, west and northern 4 mark points are equidistantly set in the upper limb of drum, as animal intake pool
Place of entry, with this 4 place of entry the water surface and bucket bottom subpoint, by the water surface and bucket part it is impartial be divided into 4 as
Limit.By requirement of experiment, platform can be arbitrarily set to the centre of a certain quadrant.Thermostatic equipment appropriate makes water temperature be maintained at 23
~25 DEG C.Water maze pond should be furnished with good water filling and drainage equipment, the position in pond once it is determined that, just not become easily
It is dynamic, especially in the test of same wheel water maze.Water maze image automatic collection and processing system can automatically pick up animal
Enter water position, swimming speed, search for the parameters such as required time, running track and search strategy of target, and can be to being acquired
Various data carry out statistics and analysis.The Experiment Training stage is carried out continuously 3d, daily training 4 times.Training when, by rat towards
Pool wall is respectively put into pond from four place of entry, and record mouse is from when entering water to needed for finding underwater hidden platform and stand thereon
Between, it as incubation period, is indicated with the second (s), after rat finds platform, allows its 10s that stands on platform.If entering 60s rat after water
Fail to find platform, then it is gently dragged to upper mounting plate, and stop 10s from water, then trained next time.Every mouse is from four
It is primary training that a place of entry, which is respectively put into pond, train twice between be spaced 30s.
Testing location: Capital University of Medical Sciences's brain major disease research center；
Testing result is as shown in Figure 6: dull-witted transgenic mice sharply declines in 1-6 days training periods, learning ability, performance
Hide plateau time under the water surface finding and increase, and after nano-packaging Q10 treatment, hence it is evident that shorten find platform when
Between.After the 7th day removes platform, the time for finding the target quadrant of original position of platform is reduced, but is obviously mentioned after nano-packaging Q10
Height stays in the time of target quadrant, illustrates that memory is significantly improved.
Examples 1 to 5 provided by the invention shows:
1) the Co-Q10 diameter of nano-packaging is smaller than space between cells, can flow with intercellular fluid, and then reaches target nerve
Member, into the deep of brain.
2) Co-Q10 of nano-packaging, which enters neuron, can improve the vigor of mitochondrial cytochrome C, enhancing ATP synthesis,
Energy is provided to neuron.
3) Co-Q10 of nano-packaging can reduce intracerebral formaldehyde, reduce formaldehyde and act on the promotion formation of senile plaque, reach
Scavenger-cell gap blocks purpose, restores the flowing of intercellular fluid.
4) Co-Q10 of nano-packaging provided by the present application does not use the external drug of synthesis, uses effective work
Property the Co-Q10 that itself just has of human nerve member, there is no toxicity to human body, and have enhancing immunocompetence.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of for treating the composition of senile dementia, which is characterized in that the component including following mass parts: 0.5~1 part auxiliary
Enzyme Q10,0.05~0.2 part of polysorbate, 0.3~0.8 part of MCT Oil, 3~8 parts of gelatin, 3~8 parts it is sweet
Oil and 3~8 parts of sorbierites.
2. composition according to claim 1, which is characterized in that the component including following mass parts: 0.7~0.85 part auxiliary
Enzyme Q10,0.08~0.12 part of polysorbate, 0.4~0.6 part of MCT Oil, 4~6 parts of gelatin, 4~6 parts it is sweet
Oil and 4~6 parts of sorbierites.
3. composition according to claim 1 or 2, which is characterized in that the composition is pulvis, the particle of the pulvis
Diameter≤50nm.
4. the preparation method of composition described in claims 1 to 3 any one, which comprises the steps of:
(1) Co-Q10, polysorbate, MCT Oil, gelatin, glycerol, sorbierite and dehydrated alcohol are mixed
It closes, obtains organic phase；
(2) organic phase is mixed with the water of 3~8 times of organic phase volumes, obtains mixed liquor；
(3) mixed liquor is heated to 60~80 DEG C, stirring is concentrated by evaporation to 0.3~0.8 times of organic phase volume, obtains nanometer
Emulsion；
(4) nanoemulsion is placed in 0~4 DEG C of environment and continues to stir, obtain a nanometer Co-Q10 suspension；
(5) by after the nanometer Co-Q10 suspension homogeneous, vacuum freeze drying, the composition for the senile dementia that obtains medical treatment.
5. the preparation method according to claim 4, which is characterized in that in the step (1) quality of Co-Q10 with it is anhydrous
The ratio between volume of ethyl alcohol is 0.5~1g:100mL.
6. the preparation method according to claim 4, which is characterized in that the rate of stirring is in the step (3) or (4)
1000~2000rpm.
7. preparation method according to claim 6, which is characterized in that the time stirred in the step (4) is 1~2h.
8. the preparation method according to claim 4, which is characterized in that the temperature of homogeneous is 45~55 in the step (5)
DEG C, the pressure of homogeneous is 200~1500bar.
9. preparation method according to claim 8, which is characterized in that the number of the homogeneous is 2~4 times, the homogeneous
Pressure gradually increase.
10. preparation method described in composition described in claims 1 to 3 any one or claim 4~9 any one is prepared into
To composition preparation treatment medicine for senile dementia in application.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101066260A (en) * | 2006-11-17 | 2007-11-07 | 姚瑶 | Coenzyme Q10 emulsion and its freeze dried prepn and their prepn process |
| CN106727441A (en) * | 2016-12-29 | 2017-05-31 | 厦门金达威生物科技有限公司 | Water-soluble nano slow-release function Co-Q10 microcapsules and preparation method and application |
-
2018
- 2018-12-28 CN CN201811622989.4A patent/CN109498596B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101066260A (en) * | 2006-11-17 | 2007-11-07 | 姚瑶 | Coenzyme Q10 emulsion and its freeze dried prepn and their prepn process |
| CN106727441A (en) * | 2016-12-29 | 2017-05-31 | 厦门金达威生物科技有限公司 | Water-soluble nano slow-release function Co-Q10 microcapsules and preparation method and application |
Non-Patent Citations (1)
| Title |
|---|
| XIFEI YANG等: "Coenzyme Q10 Reduces β-Amyloid Plaque in an APP/PS1 Transgenic Mouse Model of Alzheimer’s Disease", 《JOURNAL OF MOLECULAR NEUROSCIENCE》 * |
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