CN109395093A - A kind of diagnosis and treatment integration preparation and preparation method thereof for Alzheimer disease - Google Patents
A kind of diagnosis and treatment integration preparation and preparation method thereof for Alzheimer disease Download PDFInfo
- Publication number
- CN109395093A CN109395093A CN201811634453.4A CN201811634453A CN109395093A CN 109395093 A CN109395093 A CN 109395093A CN 201811634453 A CN201811634453 A CN 201811634453A CN 109395093 A CN109395093 A CN 109395093A
- Authority
- CN
- China
- Prior art keywords
- diagnosis
- preparation
- treatment integration
- treatment
- added
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6905—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a colloid or an emulsion
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/12—Ketones
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
- A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
- A61K49/08—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier
- A61K49/10—Organic compounds
- A61K49/12—Macromolecular compounds
- A61K49/126—Linear polymers, e.g. dextran, inulin, PEG
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
- A61K49/18—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
- A61K49/1806—Suspensions, emulsions, colloids, dispersions
- A61K49/1809—Micelles, e.g. phospholipidic or polymeric micelles
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Epidemiology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Radiology & Medical Imaging (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Dispersion Chemistry (AREA)
- Biomedical Technology (AREA)
- Neurosurgery (AREA)
- Neurology (AREA)
- Psychiatry (AREA)
- Hospice & Palliative Care (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
- General Chemical & Material Sciences (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The diagnosis and treatment integration preparation and preparation method thereof that the present invention relates to a kind of for Alzheimer disease, belongs to high molecular material and engineering in medicine technical field.Diagnosis and treatment integration preparation of the invention is the core-shell structure that amphipathic copolymer is formed, inner nuclear layer is formed by Distearoyl Phosphatidylethanolamine segment, outer shell is formed by polyethylene glycol segment, superparamagnetic nano particle is wrapped up in the inner nuclear layer and curcumin, the shell layer surface are connected with B6 small peptide.Diagnosis and treatment integration preparation of the invention integrates Brain targeting treatment of alzheimer and tracer is imaged in MRI, targeting may be implemented and be gathered in the imaging of Alzheimer disease plaque site, and the release of drug is controlled simultaneously, increase target site drug concentration and improve drug treating time, is with a wide range of applications in terms of the targeting diagnosis and treatment of Alzheimer disease and delivering drug enter brain.
Description
Technical field
The diagnosis and treatment integration preparation and preparation method thereof that the present invention relates to a kind of for Alzheimer disease, belongs to macromolecule
Material and engineering in medicine technical field.
Background technique
Alzheimer disease (Alzheimer ' s Disease, AD), is commonly called as senile dementia, is that one kind is recognized with progressive
Dysfunction and learning and memory damage the neurodegenerative disease being characterized, and are mainly caused extremely with cognitive disorder and mental act
Activity of daily living reduce even lose be clinical manifestation.With the development of aging of population, Alzheimer disease at
For an important diseases for influencing senior health and fitness and quality of life.The pathogenesis of Alzheimer disease is not yet studied clearly
Chu still lacks the drug that can effectively treat the disease.Since treatment Alzheimer disease medication need to carry out for a long time, so finding
Safe and effective drug becomes the main target for preventing and treating Alzheimer disease.
Curcumin extracts from the rhizome of Zingiber curcumin platymiscium, can be used as edible pigment use, and curcumin is not only
It can be used as food additives and fragrance, also there is certain treatment and prevention to disease.And since it is extracted from
Natural plants are increasingly becoming the hot spot of people's research with high security, the small advantage of toxic side effect.The study found that curcumin
Have the effects that anti-oxidation stress, anti-inflammatory, anticancer, and some researches show that curcumins can be used for Parkinson disease, Ah in recent years
The treatment of the neurodegenerative diseases such as Alzheimer's disease.But macromolecular drug is very low by the conevying efficiency of blood-brain barrier, medicine
Object carrier also reduces the functioning efficiency and therapeutic effect in drug cerebral ischemia region again without targeting specific.Therapeutic process at
Picture tracer (or diagnosis) cannot also carry out simultaneously, limited to the tumour containment effect for rapidly developing, shifting.Therefore, development collection target
There is highly important social effect and economic significance in the new technical means of one with imaging tracer to treatment.
The carrier that nanoparticle drug-loading system is treated as Alzheimer disease drugs can be such that chemicals pass by Brain targeting
The strategy of medicine improves the concentration of brain, reduces the adverse effect that drug generates other histoorgans, to improve therapeutic effect.
The targeted drug delivery carrier studied at present mainly has liposome, nanoparticle and micella.Wherein, micella (micelle) is amphiphilic
Property molecule acts on the pharmaceutical carrier with core-shell structure to be formed by self assembly.The hydrophobic cores of micella can contain hydrophobic
Property or insoluble drug, improve the stability of drug, and the hydrophilic fractions of micella advantageously reduce reticuloendothelial system
Phagocytosis.Targeted therapeutic carrier of the polymer micelle as Alzheimer disease, it is advantageous that can be self-assembled into divide
Son amount is more advantageous to drug through blood-brain barrier (blood-brain-barrier, BBB) within the scope of 1~15KDa.
In the past twenty years, the nanoparticle of polyethylene glycol (PEG) modification has attracted more and more preparation research persons'
Concern, with preferable biocompatibility, biodegradable and internal long circulating behavior；Especially it is worth mentioning that by more
Change class drug encapsulation to be avoided that the degradation of drug in nanoparticle, improve stability.Using polyethylene glycol as the polymerization of carrier element
Object micella is also the hot spot studied in recent years.Mu etc. screens PEG-PLGA (the polylactic acid-glycolic base second of ligand using targeting modification
Acid copolymer) micella, the targeted therapy of Alzheimer disease is realized by containing drug Flurbiprofen.The ligand of this section of targeting
Can be had on a cellular level with the TfR on specific recognition blood-brain barrier, ligand modified carrier micelle
The intake of effect, and realize to pass in the brain parenchymal cell of Flurbiprofen and release.Therefore, polyethylene glycol polymer micella be realize Ah
The good approach of Alzheimer's disease drug delivery.
Blood-brain barrier is that Erilich was first confirmed that in 1885 and proposed by Britain physiologist Hugh Davson, it
It is existing protective barrier between hematological system and brain tissue, is brain capillary (the brain capillary for having depolarising
Endothelial cells, BCEC) closely connect and compose, protection central nervous system is played a crucial role, but
Entering intracerebral transhipment to most of drug also has very big restriction effect, studies have shown that almost 98% drug molecule is all
It is difficult to penetrate and enter brain, even some polypeptides and genomic medicine, so us is given to bring on curing brain diseases
Huge challenge.And drug can need to usually have molecular weight in 500KDa or less and higher fat-soluble, just through blood-brain barrier
The drug only only a few that chemicals lane database is used to treat central system disorder at present can reach this point.So with
The development for the brain diseases that the whole world constantly deteriorates, the research of brain targeting drug delivery system are particularly significant.Currently, being passed for intracerebral
Medicine is broadly divided into such a way that Passive diffusion is through blood-brain barrier, and is mediated with receptor, transporter and absorption and penetrated blood-brain barrier
Mode.
Biology strategy is current researcher most study, mainly have it is receptor-mediated pass prescription formula, absorption
Prescription formula and transporter mediate passs prescription formula for passing of mediating.Receptor-mediated prescription formula of passing: which is most mature at present
There is how species specific receptor on brain capillary endothelial cell (BCEC) in one of Brain targeting strategy, as TfR,
LDL receptor, N- acetylcholinergic receptor and insulin receptor etc..Use ligand or the antibody of above-mentioned receptor for targeting
Molecule construction nano medicament carrying system or medicinal composition can be with the specific bindings of receptor and mediate drug enters brain.Absorption mediates
Pass prescription formula: blood-brain barrier film bear electricity, such as basement membrane side Heparan sulfate and Cavity surface side sialic acid, with sun from
The transcytosis (AMT) that absorption mediates can be caused by Electrostatic Absorption after sub- albumen contact, wherein the albumin that is cationized
It is then the representative of such cationic protein, can be used as Brain targeting functional molecular.What transporter mediated passs prescription formula: due to brain group
It knits and normal physiological function is maintained to need a large number of nutrients, and these substances can be by BBB, what is relied primarily on is exactly to turn
The mode that fortune body mediates enters brain tissue.Mainly there are amino acid transporter system, hexose transporter system, monocarboxylate transporter system
System.
For Alzheimer's disease, targeted drug delivery seems even more important.Nanoparticle drug-loading system is as alzheimer '
The carrier of silent disease targeted therapy, can make chemicals improve the concentration of brain by the strategy of brain targeting drug delivery, reduce drug pair
The adverse effect that other histoorgans generate, to improve therapeutic effect.More and more researches show that not being A β (beta amyloid
Albumen) it is only deposited on fine and close neuritic plaque and just has neurotoxicity, and the aggregation of toxicant is only real pathological change
The first step.The generation of A β and remove under physiological conditions be a dynamic equilibrium process, it is unbalance under pathologic condition to result in mind
It is lost through meta function and dull-witted.Block A beta-aggregation: the accumulation process of A β is not only by the interaction between A β peptide and peptide, together
When also by the adjusting of other albumen, so by pointedly select the Drug inhibition process, it would be possible to the target as drug
One of.Adjust the generation of A β: the effective way for inhibiting A β to generate is that selectively raising α -2 secretase activity, inhibition β, γ 2 divide
Secrete the activity of enzyme.Therefore, the adjusting for blocking and the generation of A beta-aggregation is studied, and is developed corresponding drug and be considered
It is up-and-coming.
Summary of the invention
It is provided a kind of for Alzheimer disease it is an object of the invention to overcome above-mentioned the deficiencies in the prior art place
Diagnosis and treatment integration preparation and preparation method thereof, diagnosis and treatment integration preparation collection Brain targeting Alzheimer's disease treatment of the invention with
Tracer is imaged in one in MRI, increases target site drug concentration and improves drug treating time, in the target of Alzheimer disease
It is with a wide range of applications in terms of diagnosis and treatment and delivering drug enter brain.
To achieve the above object, the technical scheme adopted by the invention is as follows: a kind of diagnosis and treatment one for Alzheimer disease
Change preparation, the diagnosis and treatment integration preparation is the core-shell structure that amphipathic copolymer is formed, and inner nuclear layer is by distearoylphosphatidyl
Ethanol amine segment is formed, and outer shell is formed by polyethylene glycol segment, and superparamagnetic nano particle and ginger are wrapped up in the inner nuclear layer
Flavine, the shell layer surface are connected with B6 small peptide.
B6 small peptide (amino acid sequence are as follows: CGHKAKGPRK) of the present invention can cross over blood-brain barrier is prepared poly- for target
It closes object nano-micelle carrier (DSPE-PEG), while superparamagnetism ferrite nano particles (SPIO) is passed through into hydrophobic electrostatic interaction
It is combined with each other with carrier assembling, finally adsorbs drug curcumin in hydrophobic inner core, synthesize carrier micelle (SPIO-DSPE-PEG/
Cur-B6), the diagnosis and treatment integration reagent with targeting Alzheimer disease is constructed.
Diagnosis and treatment integration preparation of the invention be nano-carrier, using amphipathic copolymer formed a kind of core-shell structure,
Its inner nuclear layer is formed by hydrophobicity Distearoyl Phosphatidylethanolamine segment, and outer shell is formed by hydrophilic polyglycol segment,
Hydrophobic superparamagnetic nano particle and drug is wrapped in inner nuclear layer, and B6 peptide molecule is connected to glue by amidation process
Beam water-wetted surface.Diagnosis and treatment integration preparation of the invention may be implemented targeting and be gathered in the imaging of Alzheimer disease plaque site,
And the release of drug is controlled simultaneously.
As the preferred embodiment of diagnosis and treatment integration preparation of the present invention, the partial size of the diagnosis and treatment integration preparation is
50~250nm.
As the preferred embodiment of diagnosis and treatment integration preparation of the present invention, the partial size of the superparamagnetic nano particle
For 5~30nm.
As the preferred embodiment of diagnosis and treatment integration preparation of the present invention, the amino acid sequence of the B6 small peptide is such as
Shown in SEQ ID NO:1.
As the preferred embodiment of diagnosis and treatment integration preparation of the present invention, the diagnosis and treatment integration preparation includes following
The component of parts by weight: 5~20 parts of distearoylphosphatidylethanolamine-polyethylene glycol 2000- carboxyl, superparamagnetic nano particle 1
~5 parts, 0.5~1 part of curcumin, hundred a ten thousandth of B6 small peptide to ten a ten thousandth parts.
As the preferred embodiment of diagnosis and treatment integration preparation of the present invention, the preparation of the superparamagnetic nano particle
Method are as follows: by praseodynium iron, diacetyl acetone manganese, diacetyl acetone zinc and 1, then the mixing of 2- hexadecane diol is added
Oleic acid and oleyl amine, and benzyl ether is added as solvent, argon gas protection is lower to react, and 200 DEG C of reaction 2h are then heated to 300 DEG C and return
Stream reaction 0.5h, obtains superparamagnetic nano particle.
The present invention also provides the preparation methods of above-mentioned diagnosis and treatment integration preparation, comprising the following steps:
(1) superparamagnetic nano particle is added in chloroform, and the poly- second two of Distearoyl Phosphatidylethanolamine-is added
Alcohol 2000- carboxyl, curcumin mixing, ultrasonic at room temperature, then concentration removes chloroform；
(2) it being cooled to room temperature, ultrasound, ultrafiltration is centrifuged after filtering, takes the aqueous phase solution of upper layer black transparent, it is freeze-dried,
Obtain pulverulent solids；
(3) pulverulent solids that step (2) obtains are configured to solution, 1- (3- dimethylamino-propyl) -3- ethyl is added
Carbodiimide is added n-hydroxysuccinimide and reacts after activation；Then B6 small peptide is added, reaction is overnight；
(4) reaction solution of step (3) is dialysed in distilled water, the solution after dialysis is freeze-dried to obtain diagnosis and treatment one
Change preparation.
The preferred embodiment of preparation method as diagnosis and treatment integration preparation of the present invention in the step (1), surpasses
The sound time is 10min, is concentrated as rotary evaporation after vacuumizing in 70 DEG C of water-baths.
The preferred embodiment of preparation method as diagnosis and treatment integration preparation of the present invention in the step (2), surpasses
The sound time is 15min, and the filter sizes of filtering are 220nm, sublimation drying 20h.
The preferred embodiment of preparation method as diagnosis and treatment integration preparation of the present invention, it is molten in the step (3)
The concentration of liquid is 2mg/mL, and pulverulent solids and 1- (3- dimethylamino-propyl) -3- ethyl carbodiimide, N- hydroxysuccinimidyl acyl are sub-
The mass ratio of amine is 1:2:2, activation time 10min, and the reaction time that n-hydroxysuccinimide is added is 2h；The step
(4) in, when dialysis, uses the molecular cut off of bag filter for 5000D, and it is primary that dialysis procedure 6h changes water.
Compared with prior art, the invention has the benefit that diagnosis and treatment integration preparation collection Brain targeting A Er of the invention
The treatment of Ci Haimo disease, in one, may be implemented targeting and be gathered in the imaging of Alzheimer disease plaque site with MRI imaging tracer, and
The release of drug is controlled simultaneously, increase target site drug concentration and improves drug treating time, in Alzheimer disease
Targeting diagnosis and treatment and delivering drug are with a wide range of applications in terms of entering brain.
Detailed description of the invention
Fig. 1 is the transmission electron microscope photo figure of superparamagnetic nano particle.
Fig. 2 is the transmission electron microscope photo figure of diagnosis and treatment integration preparation prepared by embodiment 1.
Fig. 3 is the transmission electron microscope photo figure of diagnosis and treatment integration preparation prepared by embodiment 2.
Fig. 4 is the transmission electron microscope photo figure of preparation prepared by comparative example 1.
Fig. 5 is the transmission electron microscope photo figure of preparation prepared by comparative example 2.
Fig. 6 is the MRI-T2WI image in effect example 1.
Fig. 7 is the escape incubation period result figure in effect example 2.
Specific embodiment
Purposes, technical schemes and advantages in order to better illustrate the present invention, below in conjunction with the drawings and specific embodiments pair
The present invention is described further.
Test method employed in the embodiment of the present invention is conventional method unless otherwise specified；Used material,
Reagent etc., is commercially available unless otherwise specified, wherein B6 small peptide derives from Shanghai gill biochemistry Co., Ltd.
In the embodiment of the present invention, superparamagnetic nano particle the preparation method is as follows:
By 2mmol Fe (acac)3(praseodynium iron), 0.6mmol Mn (acac)2(diacetyl acetone manganese),
0.4mmol Zn(acac)2(diacetyl acetone zinc) and 10mmol 1,2- hexadecane diol, which is fitted into reaction vessel, to be mixed, then is divided
Not Jia Ru 3mmol oleic acid and 3mmol oleyl amine, and 10mL benzyl ether is added and makees solvent, magnetic agitation, is heated under protection of argon gas
200 DEG C of heat preservation 2h, are then heated to 300 DEG C of back flow reaction 0.5h.After being cooled to room temperature, 40mL ethyl alcohol is added in the reaction product
Centrifugation (8000rpm, 10min) obtains the sediment of brownish black afterwards, removes filtrate, be added into precipitating 0.05mL oleic acid and
After 0.05mL oleamide, sediment is dissolved with n-hexane and is dispersed.Centrifugation (8000rpm, 10min) removes undispersed miscellaneous afterwards
Matter, the again precipitating centrifugation (8000rpm, 10min) in dehydrated alcohol, is finally dispersed in n-hexane and saves.
The particle diameter distribution of the present embodiment particle is measured with dynamic laser light scattering instrument, Fig. 1 is superparamagnetic nano particle
Transmission electron microscope photo figure.As shown in Figure 1, superparamagnetic nanoparticle average grain diameter be 8~10nm, particle it is spherical in shape and
It is uniformly dispersed.
Embodiment 1
One embodiment of diagnosis and treatment integration preparation of the present invention, preparation method are as follows:
(1) superparamagnetic nano particle of 1 part of components by weight percent of partial size about 5nm is weighed, 5mL chloroform is added, will mix
Close object and 5 parts of components by weight percent of distearoylphosphatidylethanolamine-polyethylene glycol 2000- carboxyl and 0.5 part of components by weight percent of ginger
After flavine mixing, ultrasound 10min at room temperature keeps polymer and magnetic nano-particle fully dispersed and dissolve, by round bottom after mixing
Flask is placed on Rotary Evaporators, 70 DEG C of water-bath, is evacuated to rotary evaporation after vacuum；By hydrophobic effect, modified in particle surface
Oleic acid alkyl chain on coated single layer there is preferable water-soluble DSPE-PEG2000 phospholipid molecule, when organic solvent evaporation is dangerous
After to the greatest extent, nanostructure mutually successfully goes to water phase by oily；
(2) after it is cooled to room temperature, ultrasonic probe effect 15min makes its dispersion；The ultrafiltration again after 220nm membrane filtration
Centrifugation removes bottom sediment, takes the solution of the water phase nanostructure of upper layer black transparent, is then freeze-dried 20 hours, obtains
To pulverulent solids；
(3) pulverulent solids for obtaining step (2) prepare micellar solution 2.5mL, and 2 times of weight of pulverulent solids are added
EDC (1- (3- dimethylamino-propyl) -3- ethyl carbodiimide) activates the NHS of addition 2 times of weight of pulverulent solids after 10min
(n-hydroxysuccinimide) reacts 2h, components by weight percent parts per million B6 small peptide is then added, reaction is overnight；
(4) it is dialysed in distilled water with the bag filter of 5000D and removes free B6 small peptide, it is primary that 6h changes water；By micella water
Solution is freeze-dried to obtain diagnosis and treatment integration preparation of the invention.
The particle diameter distribution of the present embodiment preparation is measured with dynamic laser light scattering instrument, Fig. 2 is the present embodiment diagnosis and treatment integration
The transmission electron microscope photo figure of preparation.As shown in Figure 2, the average grain diameter of the present embodiment is 102nm, and particle is spherical in shape, more
Dispersibility is 0.215, is dispersed relatively uniform.
Embodiment 2
One embodiment of diagnosis and treatment integration preparation of the present invention, preparation method are as follows:
(1) superparamagnetic nano particle of 5 parts of components by weight percent of partial size about 5nm is weighed, 5mL chloroform is added, will mix
Close object and 20 parts of components by weight percent of distearoylphosphatidylethanolamine-polyethylene glycol 2000- carboxyl and 1 part of components by weight percent of turmeric
After element mixing, ultrasound 10min at room temperature keeps polymer and magnetic nano-particle fully dispersed and dissolve, burns round bottom after mixing
Bottle is placed on Rotary Evaporators, 70 DEG C of water-bath, is evacuated to rotary evaporation after vacuum；By hydrophobic effect, in particle surface modification
Single layer has been coated on oleic acid alkyl chain has preferable water-soluble DSPE-PEG2000 phospholipid molecule, when organic solvent evaporation totally
Afterwards, nanostructure mutually successfully goes to water phase by oily；
(2) after it is cooled to room temperature, ultrasonic probe effect 15min makes its dispersion；The ultrafiltration again after 220nm membrane filtration
Centrifugation removes bottom sediment, takes the solution of the water phase nanostructure of upper layer black transparent, is then freeze-dried 20 hours, obtains
To pulverulent solids；
(3) pulverulent solids for obtaining step (2) prepare micellar solution 2.5mL, and 2 times of weight of pulverulent solids are added
EDC (1- (3- dimethylamino-propyl) -3- ethyl carbodiimide) activates the NHS of addition 2 times of weight of pulverulent solids after 10min
(n-hydroxysuccinimide) reacts 2h, ten a ten thousandth part B6 small peptide of components by weight percent is then added, reaction is overnight；
(4) it is dialysed in distilled water with the bag filter of 5000D and removes free B6 small peptide, it is primary that 6h changes water；By micella water
Solution is freeze-dried to obtain diagnosis and treatment integration preparation of the invention.
The particle diameter distribution of the present embodiment preparation is measured with dynamic laser light scattering instrument, Fig. 3 is the present embodiment diagnosis and treatment integration
The transmission electron microscope photo figure of preparation.The average grain diameter of the present embodiment is 113nm, and particle is spherical in shape, polydispersity
0.228, disperse relatively uniform.
Comparative example 1
The superparamagnetic nano particle of 50mg partial size about 25nm is weighed, 5mL chloroform is added, by mixture and 50mg bis-
After the mixing of stearyl phosphatidyl ethanol amine-polyethylene glycol 2000-carboxyl, ultrasound 10min, makes polymer and magnetic Nano at room temperature
Particle is fully dispersed and dissolves, and round-bottomed flask is placed on Rotary Evaporators after mixing, 70 DEG C of water-bath, rotates steaming after being evacuated to vacuum
Hair；By hydrophobic effect, single layer has been coated on the oleic acid alkyl chain of particle surface modification has preferable water-soluble DSPE-
PEG2000 phospholipid molecule.When organic solvent evaporation totally after, nanostructure mutually successfully goes to water phase by oily.Room is cooled to it
Wen Hou, ultrasonic probe effect 15min make its dispersion, and ultrafiltration is centrifuged again after 220nm membrane filtration, is removed bottom sediment, is taken
Then the solution of the water phase nanostructure of upper layer black transparent is freeze-dried 20 hours, obtains pulverulent solids.
The particle diameter distribution of this comparative example preparation is measured with dynamic laser light scattering instrument, Fig. 4 is the transmission of this comparative example preparation
Electron micrograph figure.As shown in Figure 4, average grain diameter 110nm, particle is spherical in shape, polydispersity 0.208, dispersion ratio
It is more uniform.
Comparative example 2
The superparamagnetic nano particle of 50mg partial size about 25nm is weighed, 5mL chloroform is added, by mixture and 50mg bis-
After stearyl phosphatidyl ethanol amine-polyethylene glycol 2000-carboxyl and the mixing of 30mg curcumin, ultrasound 10min, makes to polymerize at room temperature
Object and magnetic nano-particle it is fully dispersed and dissolution, round-bottomed flask is placed on Rotary Evaporators after mixing, 70 DEG C of water-bath, is evacuated to
Rotary evaporation after vacuum；By hydrophobic effect, single layer has been coated on the oleic acid alkyl chain of particle surface modification with preferable water
The DSPE-PEG2000 phospholipid molecule of dissolubility.When organic solvent evaporation totally after, nanostructure mutually successfully goes to water phase by oily.To
After it is cooled to room temperature, ultrasonic probe effect 15min makes its dispersion.Ultrafiltration is centrifuged sample again after 220nm membrane filtration, removal
Bottom sediment takes the solution of the water phase nanostructure of upper layer black transparent, is then freeze-dried 20 hours, obtains powdered solid
Body.
The particle diameter distribution of this comparative example preparation is measured with dynamic laser light scattering instrument, Fig. 5 is this comparative example diagnosis and treatment integration
The transmission electron microscope photo figure of preparation, TEM photo show that the average grain diameter of nano particle is 132nm, and particle is spherical in shape, more
Dispersibility is 0.235, is dispersed relatively uniform.
The internal MRI development effect of 1 embodiment of the present invention of effect example, 1 preparation
After groups of animals chloral hydrate anesthesia, the connection dedicated nuclear magnetic scanning four-way phased-array coil of toy is (straight
Diameter 5cm) MRI scan is carried out, signal evolution after 2,6,8 weeks drugs of injection is observed on MRI routine sequence；And use ROI
Technology measures the T2 signal strength of drug, calculates relaxation rate R2, calculates the hippocampus area R2 increment rate before and after injection drug.
After injecting nano material, MRI-T2WI image is as shown in Figure 6.It will be appreciated from fig. 6 that there is strip of sheet, spot in inside tumor
Sheet low signal area, and the signal for injecting the nude mice of targeted nano material (embodiment 1) is (more right without targeted nano material than injecting
Ratio 1) nude mice it is high by 12%.Show that hippocampus of mice body region tissue can be gathered in by targeting diagnosis and treatment preparation, has good aobvious
Shadow function, and target the targeting of diagnosis and treatment preparation more preferably.The therapeutic effect of 2 embodiment of the present invention of effect example, 2 preparation
APPswe/PS1dE9 double transgenic mouse model is established, to evaluate diagnosis and treatment of the nano material to Alzheimer disease
Effect；The Spatial learning and memory ability of mouse is evaluated by water maze laboratory (Morris water maze, MWM).Mouse exists
Laboratory rearing one week to adapt to environment.Water maze is carried out after a week and adapts to test, prepares determined with Morris water device, mouse is put into
It is wherein adapted to, is not lauched again with staying in 10 seconds on platform as standard, every detection in mouse one day three times, excludes platform
Place quadrant is put into mouse the centre of other three quadrants is adherent.It appears on the stage 10 seconds to be no longer lauched with mouse and thinks once to survey
The completion of examination.Photograph the swimming process of mouse.According to the video recording taken the photograph, the behavior of mouse is divided with determined with Morris water software
Analysis.The difference of different mouse Spatial memory abilities is judged according to the time is appeared on the stage.By observing and recording mouse association in water
Time, the strategy of use and their swimming track needed for swimming in case and finding the underwater escape platform of plant, analyze and push away
The ability of study, memory and the spatial cognition of disconnected animal etc..The mouse after different experiments group material processing is analyzed, in water
Escape incubation period in maze experiment evaluates the promotion effect of the mouse Spatial learning and memory ability of nano-medicament carrier pair.
It is small that diagnosis and treatment integration preparation prepared by blank control group, comparative example 2 and embodiment 2 is applied to APP/PS1 transgenosis
The escape incubation period result of water maze laboratory is as shown in Figure 7 after mouse.As shown in Figure 7, the results showed that targeting diagnosis and treatment preparation (embodiment
2) there is castering action to the Spatial learning ability of mouse, and effect is better than without targeting diagnosis and treatment preparation (comparative example 2).
Finally, it should be noted that the above embodiments are merely illustrative of the technical solutions of the present invention rather than protects to the present invention
The limitation of range is protected, although the invention is described in detail with reference to the preferred embodiments, those skilled in the art should
Understand, it can be with modification or equivalent replacement of the technical solution of the present invention are made, without departing from the essence of technical solution of the present invention
And range.
SEQUENCE LISTING
<110>Guangzhou Chuan Sai bio-medical material Co., Ltd
<120>a kind of diagnosis and treatment integration preparation and preparation method thereof for Alzheimer disease
<130> 20181120
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 10
<212> PRT
<213>artificial synthesized
<400> 1
Cys Gly His Lys Ala Lys Gly Pro Arg Lys
1 5 10
Claims (10)
1. a kind of diagnosis and treatment integration preparation for Alzheimer disease, which is characterized in that the diagnosis and treatment integration preparation is two
The core-shell structure that parent's property copolymer is formed, inner nuclear layer are formed by Distearoyl Phosphatidylethanolamine segment, and outer shell is by poly- second two
Alcohol segment is formed, and superparamagnetic nano particle and curcumin, the shell layer surface and B6 small peptide phase are wrapped up in the inner nuclear layer
Connection.
2. diagnosis and treatment integration preparation as described in claim 1, which is characterized in that the partial size of the diagnosis and treatment integration preparation is 50
~250nm.
3. diagnosis and treatment integration preparation as described in claim 1, which is characterized in that the partial size of the superparamagnetic nano particle is
5~30nm.
4. diagnosis and treatment integration preparation as described in claim 1, which is characterized in that the amino acid sequence such as SEQ of the B6 small peptide
Shown in ID NO:1.
5. diagnosis and treatment integration preparation as described in claim 1, which is characterized in that the diagnosis and treatment integration preparation includes following heavy
Measure the component of part: 5~20 parts of distearoylphosphatidylethanolamine-polyethylene glycol 2000- carboxyl, superparamagnetic nano particle 1~5
Part, 0.5~1 part of curcumin, ten a ten thousandth of B6 small peptide to parts per million.
6. diagnosis and treatment integration preparation as described in claim 1, which is characterized in that the preparation side of the superparamagnetic nano particle
Method are as follows: by praseodynium iron, diacetyl acetone manganese, diacetyl acetone zinc and 1, then oil is added in the mixing of 2- hexadecane diol
Acid and oleyl amine, and benzyl ether is added as solvent, argon gas protection is lower to react, and 200 DEG C of reaction 2h are then heated to 300 DEG C of reflux
0.5h is reacted, superparamagnetic nano particle is obtained.
7. the preparation method of diagnosis and treatment integration preparation as described in any one of claims 1 to 6, which is characterized in that including following
Step:
(1) superparamagnetic nano particle is added in chloroform, and distearoylphosphatidylethanolamine-polyethylene glycol is added
2000- carboxyl, curcumin mixing, ultrasonic at room temperature, then concentration removes chloroform；
(2) it is cooled to room temperature, ultrasound, ultrafiltration is centrifuged after filtering, takes the aqueous phase solution of upper layer black transparent, is freeze-dried, is obtained
Pulverulent solids；
(3) pulverulent solids that step (2) obtains are configured to solution, 1- (3- dimethylamino-propyl) -3- ethyl carbon two is added
Imines is added n-hydroxysuccinimide and reacts after activation；Then B6 small peptide is added, reaction is overnight；
(4) reaction solution of step (3) is dialysed in distilled water, the solution after dialysis is freeze-dried to obtain diagnosis and treatment integration system
Agent.
8. the preparation method of diagnosis and treatment integration preparation as claimed in claim 7, which is characterized in that in the step (1), ultrasound
Time is 10min, is concentrated as rotary evaporation after vacuumizing in 70 DEG C of water-baths.
9. the preparation method of diagnosis and treatment integration preparation as claimed in claim 7, which is characterized in that in the step (2), ultrasound
Time is 15min, and the filter sizes of filtering are 220nm, sublimation drying 20h.
10. the preparation method of diagnosis and treatment integration preparation as claimed in claim 7, which is characterized in that molten in the step (3)
The concentration of liquid is 2mg/mL, and pulverulent solids and 1- (3- dimethylamino-propyl) -3- ethyl carbodiimide, N- hydroxysuccinimidyl acyl are sub-
The mass ratio of amine is 1:2:2, activation time 10min, and the reaction time that n-hydroxysuccinimide is added is 2h；The step
(4) in, when dialysis, uses the molecular cut off of bag filter for 5000D, and it is primary that dialysis procedure 6h changes water.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811634453.4A CN109395093A (en) | 2018-12-29 | 2018-12-29 | A kind of diagnosis and treatment integration preparation and preparation method thereof for Alzheimer disease |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811634453.4A CN109395093A (en) | 2018-12-29 | 2018-12-29 | A kind of diagnosis and treatment integration preparation and preparation method thereof for Alzheimer disease |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109395093A true CN109395093A (en) | 2019-03-01 |
Family
ID=65462793
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811634453.4A Pending CN109395093A (en) | 2018-12-29 | 2018-12-29 | A kind of diagnosis and treatment integration preparation and preparation method thereof for Alzheimer disease |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109395093A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110507830A (en) * | 2019-09-29 | 2019-11-29 | 中山大学肿瘤防治中心（中山大学附属肿瘤医院、中山大学肿瘤研究所） | A kind of nano-probe and its preparation for Alzheimer disease pathogenic protein |
| CN110559454A (en) * | 2019-09-29 | 2019-12-13 | 中山大学孙逸仙纪念医院 | nano composite medicine for diagnosing and treating Alzheimer's disease |
| CN111135314A (en) * | 2020-02-17 | 2020-05-12 | 中山大学附属第三医院 | Nano-composite for early diagnosis and treatment of gastric cancer and preparation method thereof |
| CN113311153A (en) * | 2021-05-12 | 2021-08-27 | 华中科技大学 | Multifunctional nanoparticle for integrated diagnosis and treatment of Alzheimer disease |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103449530A (en) * | 2013-09-06 | 2013-12-18 | 南京东纳生物科技有限公司 | Preparation method of high-performance magnetic manganese zinc ferrite nanostars and nanoclusters |
| CN104623658A (en) * | 2014-12-29 | 2015-05-20 | 上海师范大学 | Water-soluble ferrate composite nano-particle as well as preparation method and application thereof |
| US20160346389A1 (en) * | 2013-09-12 | 2016-12-01 | Albert Einstein College Of Medicine Inc. | Modified paramagnetic nanoparticles for targeted delivery of therapeutics and methods thereof |
-
2018
- 2018-12-29 CN CN201811634453.4A patent/CN109395093A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103449530A (en) * | 2013-09-06 | 2013-12-18 | 南京东纳生物科技有限公司 | Preparation method of high-performance magnetic manganese zinc ferrite nanostars and nanoclusters |
| US20160346389A1 (en) * | 2013-09-12 | 2016-12-01 | Albert Einstein College Of Medicine Inc. | Modified paramagnetic nanoparticles for targeted delivery of therapeutics and methods thereof |
| CN104623658A (en) * | 2014-12-29 | 2015-05-20 | 上海师范大学 | Water-soluble ferrate composite nano-particle as well as preparation method and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| KWOK KIN CHENG ET AL.: "Curcumin-conjugated magnetic nanoparticles for detecting amyloid plaques in Alzheimer"s disease mice using magnetic resonance imaging (MRI)", 《BIOMATERIALS》 * |
| ZHONGYANG LIU ET AL.: "B6 Peptide-Modified PEG-PLA Nanoparticles for Enhanced Brain Delivery of Neuroprotective Peptide", 《BIOCONJUGATE CHEM.》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110507830A (en) * | 2019-09-29 | 2019-11-29 | 中山大学肿瘤防治中心（中山大学附属肿瘤医院、中山大学肿瘤研究所） | A kind of nano-probe and its preparation for Alzheimer disease pathogenic protein |
| CN110559454A (en) * | 2019-09-29 | 2019-12-13 | 中山大学孙逸仙纪念医院 | nano composite medicine for diagnosing and treating Alzheimer's disease |
| CN110559454B (en) * | 2019-09-29 | 2022-04-01 | 中山大学孙逸仙纪念医院 | Nano composite medicine for diagnosing and treating Alzheimer's disease |
| CN111135314A (en) * | 2020-02-17 | 2020-05-12 | 中山大学附属第三医院 | Nano-composite for early diagnosis and treatment of gastric cancer and preparation method thereof |
| CN113311153A (en) * | 2021-05-12 | 2021-08-27 | 华中科技大学 | Multifunctional nanoparticle for integrated diagnosis and treatment of Alzheimer disease |
| CN113311153B (en) * | 2021-05-12 | 2023-05-26 | 华中科技大学 | Multifunctional nanoparticle for diagnosis and treatment of Alzheimer disease |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Guo et al. | Research on the fate of polymeric nanoparticles in the process of the intestinal absorption based on model nanoparticles with various characteristics: size, surface charge and pro-hydrophobics | |
| Bourquin et al. | Biodistribution, clearance, and long‐term fate of clinically relevant nanomaterials | |
| CN109395093A (en) | A kind of diagnosis and treatment integration preparation and preparation method thereof for Alzheimer disease | |
| Yang et al. | Chitosan mediated solid lipid nanoparticles for enhanced liver delivery of zedoary turmeric oil in vivo | |
| Wilson et al. | Chitosan nanoparticles as a new delivery system for the anti-Alzheimer drug tacrine | |
| US20080311182A1 (en) | Multistage delivery of active agents | |
| Cui et al. | Dual-target peptide-modified erythrocyte membrane-enveloped PLGA nanoparticles for the treatment of glioma | |
| Chaurasia et al. | Potential of cationic-polymeric nanoparticles for oral delivery of naringenin: in vitro and in vivo investigations | |
| Bondì et al. | Lipid nanoparticles for drug targeting to the brain | |
| Song et al. | Surface-modified PLGA nanoparticles with PEG/LA-chitosan for targeted delivery of arsenic trioxide for liver cancer treatment: Inhibition effects enhanced and side effects reduced | |
| EP2029119A2 (en) | Functionalized solid polymer nanoparticles for diagnostic and therapeutic applications | |
| EP2848262B1 (en) | Cell-specific targeting by nanostructured carrier systems | |
| CN108126189B (en) | Nano composite carrier drug delivery system assembled in grading manner and application thereof | |
| Liu et al. | Combined strategies of apomorphine diester prodrugs and nanostructured lipid carriers for efficient brain targeting | |
| Andreozzi et al. | Size-stable solid lipid nanoparticles loaded with Gd-DOTA for magnetic resonance imaging | |
| Lee et al. | Low‐Density Lipoprotein‐Mimicking Nanoparticles for Tumor‐Targeted Theranostic Applications | |
| Navarro et al. | Biodistribution of PLGA and PLGA/chitosan nanoparticles after repeat-dose oral delivery in F344 rats for 7 days | |
| Verma et al. | Nanoparticles: A novel approach to target tumors | |
| Modak et al. | Magnetic nanostructure-loaded bicontinuous nanospheres support multicargo intracellular delivery and oxidation-responsive morphological transitions | |
| Wang et al. | Efficient exosome extraction through the conjugation of superparamagnetic iron oxide nanoparticles for the targeted delivery in rat brain | |
| Farhangi et al. | Peptide mediated targeted delivery of gold nanoparticles into the demyelination site ameliorates myelin impairment and gliosis | |
| Ekhator et al. | Advances and opportunities in nanoparticle drug delivery for central nervous system disorders: a review of current advances | |
| Saeedi et al. | Passive targeted drug delivery of venlafaxine hcl to the brain by modified chitosan nanoparticles: Characterization, cellular safety assessment, and in vivo evaluation | |
| Lekshmi et al. | Sub acute toxicity assessment of glipizide engineered polymeric nanoparticles | |
| Quarta et al. | Multilayered magnetic nanobeads for the delivery of peptides molecules triggered by intracellular proteases |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190301 |
|
| RJ01 | Rejection of invention patent application after publication |