CN106344513A - Mitochondrion-targeted micelle material and curcumin micelle preparation prepared from material - Google Patents
Mitochondrion-targeted micelle material and curcumin micelle preparation prepared from material Download PDFInfo
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Abstract
The invention relates to a mitochondrion-targeted micelle material which is formed by bonding one hydroxyl on a hyaluronate oligomer with 5-carboxyl of (5-carboxylamyl)-triphenyl phosphonium bromide through an ester bond. The general formula is disclosed in the specification, wherein n is a positive integer, and the molecular weight of the hyaluronate oligomer is less than 10000 Da. The invention also relates to a preparation method of the mitochondrion-targeted micelle material, and a curcumin micelle preparation prepared from the micelle material and a preparation method thereof. The micelle material has tumor and mitochondrion targeting properties; and the curcumin micelle preparation prepared from the micelle material can precisely guide the curcumin into the tumor cells and mitochondrions thereof, thereby better displaying the antitumor function.
Description
Technical field
A kind of the present invention relates to target medicine carrier and formulation art, more specifically it relates to Mitochondrially targeted micellar material
And the curcumin micellar preparation with the preparation of this material.
Background technology
Malignant tumor is a kind of commonly encountered diseases and frequently-occurring disease, affects our quality of life, is our health " killer ".From
Just the frightened degree to it for the people is can be seen that in common saying " talking cancer complexion changed ".In case of human, the mortality rate that malignant tumor causes
Occupy second in the mortality rate that all diseases cause.How to solve this thorny social common problem of tumor, be people's mesh
The severeest problem above faced.
Varied to the Therapeutic Method of tumor at this stage, medicine is also varied, and here just no longer one by one
Repeat.Emphasis says Drug therapy, and Drug therapy is the common method of oncotherapy, and Drug therapy compares other therapies one
Fixed advantage, but Drug therapy itself also has certain defect, and existing anticarcinogen does not almost have to tumor tissues and cell
Selectivity, thus generally existing toxicity is big, curative effect is low, patient's poor compliance the shortcomings of, and tumor cell is also easy to produce drug resistance,
So antitumor drug is difficult to play curative effect, is difficult to for patients reach therapeutic purposes, breaks away from slight illness.Therefore, prepare
A kind of medicine, it can play antineoplastic drug effect, specifically target tumor tissue or cell can just become people simultaneously again
The direction that considers.
In numerous antitumor drug, administration nano-drug administration system is shown one's talent by its own advantage.Nano-carrier particle diameter is relatively
Little, be conducive to keeping the stability of medicine itself, the nano-micelle of certain particle diameter has epr effect, has passive targeting.Targeting
Delivery system (targeted drug delivery system, tdds) is selectivity by medicine positioning or to be enriched with
Medicament carrier system in a certain specific site.Target medicine preparation can rely on the particle diameter of its carrier, surface nature or
Pharmaceutical carrier can produce specific action with particular organization, or can be in response to some physically or chemically conditions by pharmaceutical carrier
The particularity of release medicine etc. makes drug-rich.
Curcumin (curcumin, cur) is a kind of phenols chemical combination extracting from the plant of Zingiberaceae and Araeceae class
Thing, has the activity of uniqueness.Curcumin, orange-yellow crystalline powder, is in yellow when acid, is in bronzing when alkaline, molecular formula
c21h20o6, molecular weight 368.37, water insoluble and ether, it is dissolved in ethanol, propylene glycol, acetone, be soluble in glacial acetic acid, its structural formula
For:
Medical research shows, curcumin mainly has multiple effect such as function of gallbladder promoting, blood fat reducing, anticoagulant, antioxidation, anticancer.At present
The antitumor machanism of known curcumin is various and complicated.Briefly, curcumin antitumor mainly passes through to suppress tumor thin
Intracellular growth, propagation, the migration of suppression tumor cell, inducing apoptosis of tumour cell, increase the chemosensitivity of tumor cell, retardance
The cycle of cell is in g2/m phase etc..This composition of curcumin, it has no obvious toxic and side effects while playing therapeutical effect.
Curcumin antitumor action is relatively broad, and recently, increasing people pays close attention to curcumin, studies its antitumor action and machine
Reason.
Although Rhizoma Curcumae Longae have plurality of advantages, curcumin people in water can not dissolve, all sensitive to light, heat, fast light
Property and thermostability poor, these shortcomings hinder curcumin play anti-tumor activity, also constrain it in field of medicaments simultaneously
Application.Therefore, the suitable pharmaceutical dosage form of developmental research.
Content of the invention
For solving problem above, the invention provides a kind of Mitochondrially targeted micellar material, it is by oligomerization hyaluronic acid
A hydroxyl be formed by connecting by ester bond with the 5- carboxyl of (5- carboxy pentyl)-triphenylphosphinebromide, its formula is:
Wherein, n is positive integer, and is the molecular weight of described oligomerization hyaluronic acid less than 10000da.
Present invention also offers a kind of preparation method of above-mentioned Mitochondrially targeted micellar material, by make hyaluronate sodium with
(5- carboxy pentyl)-triphenylphosphinebromide reacts under the catalytic action of dmap, edc and obtains described Mitochondrially targeted micelle material
Material.
Further, the method comprising the steps of:
1) prepare the aqueous solution of hyaluronate sodium；
2) dmso solution will in (5- carboxy pentyl)-triphenylphosphinebromide, dmap, edc solution dmso, be prepared into, and in
Stir 1 hour at 60 DEG C；
3) described dmso solution is added dropwise in described aqueous solution, and is added thereto to tetrabutyl ammonium bromide, in room temperature
Under stir and terminate to reaction, the hyaluronate sodium being added with the weight ratio of (5- carboxy pentyl)-triphenylphosphinebromide is
0.00882:0.02079；
4) by step 3) reaction product solution that obtains is transferred to bag filter, dialyses in deionized water, to removing dms0；
5) the reaction product solution lyophilization through dialysis, that is, obtain described Mitochondrially targeted micellar material.
Present invention also offers a kind of Mitochondrially targeted curcumin micellar preparation, it wraps up Rhizoma Curcumae Longae by above-mentioned micellar material
Element forms.
Present invention also offers a kind of preparation method of above-mentioned Mitochondrially targeted curcumin micellar preparation, walk including following
Rapid:
1) above-mentioned micellar material and curcumin are dissolved in equipped with the container of solvent；
2) rotary evaporation removes described solvent, forms medicine film skeleton to described container inner wall；
3) described container is placed in oil bath, and adds water in described container, so that medicine film is solidified；
4) ultrasonic aquation；
5) filtering with microporous membrane, obtains described curcumin micellar preparation.
Preferably, the weight of described micellar material and described curcumin is than for 30:1-50:1.
Preferably, described solvent is acetone.
Preferably, step 3) in solidify described medicine film temperature be 40-80 DEG C.
Preferably, step 4) in ultrasonic aquation time be 5-15min.
Preferably it is characterised in that described preparation method comprises the following steps:
1) the described micellar material than 30:1-50:1 for the weight and curcumin are dissolved in acetone and make acetone soln, load
In container；
2) acetone described in rotary evaporation, forms medicine film skeleton to described container inner wall；
3) described container is placed in 80 DEG C of oil baths, is added dropwise over about 40 DEG C of water, and add little magneton so that medicine film is solidified；
4) ultrasonic aquation 15min；
5) 0.45 μm of filtering with microporous membrane, obtains described curcumin micellar preparation.
By micellar material and the micellar preparation of the present invention, can be by accurate for curcumin target tumor, prepared Rhizoma Curcumae Longae
Plain micellar preparation has the advantages that envelop rate height, particle diameter are little, and also has obvious slow releasing function.
Brief description
Fig. 1 is hyaluronic acid structure h1- nmr spectrogram；
Fig. 2 is micellar material structure h of the present invention1- nmr spectrogram；
Fig. 3 is the linear of directrix curve and regression equation；
Fig. 4 is blank micella grain size distribution；
Fig. 5 is curcumin micelle grain size distribution；
The micelle grain size distribution that Fig. 6 obtains for ultrasonic time 5min；
The micelle grain size distribution that Fig. 7 obtains for ultrasonic time 10min；
The micelle grain size distribution that Fig. 8 obtains for ultrasonic time 15min；
Fig. 9 is the micelle grain size distribution that micellar material and curcumin ratio obtain for 30:1；
Figure 10 is the micelle grain size distribution that micellar material and curcumin ratio obtain for 40:1；
Figure 11 is the micelle grain size distribution that micellar material and curcumin ratio obtain for 50:1；
The micelle grain size distribution that Figure 12 obtains for 40 DEG C of solidification temperature；
The micelle grain size distribution that Figure 13 obtains for 60 DEG C of solidification temperature；
The micelle grain size distribution that Figure 14 obtains for 80 DEG C of solidification temperature；
Specific embodiment
Below in conjunction with accompanying drawing, the principle of the present invention and feature are described, example is served only for explaining the present invention, and
Non- for limiting the scope of the present invention.
Polymer micelle (polymericmicelles) is to be formed by amphipathic block polymerization copolymer, and it is a kind of
Thermodynamically stable system.In forming process, the hydrophobic section of copolymer is subject to h2The exclusion of o molecule, automatically associating to assemble forms
The hydrophobic inner core of this microparticle dispersion system, and its close h2O section then forms the close h of this material2O outer layer makes it in h2Steady in o
Fixed.
Polymer micelle belongs to a kind of scattered system of microgranule, itself has the incomparable part of a lot of other materials.Particle diameter
Less, degree of scatter is big, can allow be difficult to dissolve medicine and reaches more preferable dissolution degree, accelerates its dissolving speed in media as well
Rate, makes the medicine of Sq as far as possible be present in human body；Size is different, and distribution situation in vivo is different；Medicine quilt
Carrier contains, and with the volatilization of blocking drugs bad smell, can increase the compliance of patient to a certain extent；Medicine can be made simultaneously as far as possible
The physical property of thing, chemical property do not occur acutely to change, and keep stable state；The carrier containing medicine is varied, property
Matter is also different, so medicine preparation can be become targeted drug, slow releasing pharmaceutical, controlled release drug etc..Current research shows, different
The polymer latex bundle nature of carrier is different, and the mode of release and the principle of release are also different.After micelle is decomposed, inner
Face medicine is along with just revealing outside；The factor that impact micelle plays a role in vivo has a lot, and such as polymeric material belongs to
In which kind of, it have which composition and it temperature under conditions etc..
Obtain polymer micelle generally can be divided into three classes, and we often use dialysis and thin film dispersion in laboratory
Method both methods.Because dialysis ratio is relatively time-consuming and relatively costly using bag filter in dialysis, so this test adopts
Film dispersion method prepares micelle.Film dispersion method principle fairly simple it is simply that polymer carrier materials (in the such as present invention use
Hyaluronate sodium and (5- carboxy pentyl)-triphenylphosphinebromide) and medicine (curcumin used in such as this test) use
Suitable organic solvent dissolving, then uses Rotary Evaporators rotary evaporation, it is required that removing organic solvent has just obtained us
Film.
There is much Mitochondrially targeted medicine at present, species is not also single.Wherein, most representational is that electronics moves
Position lipophilic cation (delocalizedlipophilic cations, dlcs) compound, attempts in the present invention using (5-
Carboxy pentyl)-triphenylphosphine cation, as the lipophilic cation of electronics displacement, it can pass through line grain by transmembrane potential
The cell membrane of body.Tumor cell is different with the transmembrane potential on normal cell surface, and Comparatively speaking, the transmembrane potential of tumor cell surface is high
Some.Cell surface transmembrane potential higher it is possible to provide more power for lipophilic cation, just have more cationes and enter
Mitochondrion.So, tumor cell makes more Mitochondrially targeted molecule enter in cell because cell surface transmembrane potential is high.
In an experiment, (5- carboxy pentyl)-triphenylphosphinebromide, as described micelle material are connected on hyaluronic acid by esterification
Material, this micellar material has amphipathic, then passes through film dispersion method with this micellar material and prepare to contain the glue of curcumin
Bundle.In prepared micelle, hyaluronic acid has tumor-targeting, and (5- carboxy pentyl) triphenylphosphinebromide has mitochondrion target
Tropism, curcumin has antitumor action.Thus obtain the antitumor drug of new double targeting.
Based on this, the present invention is prepared for a kind of new micellar material, its both can targets neoplastic cells, again can targeting line grain
Body, concrete scheme is described in detail below.
1. the synthesis of micellar material
Inventor attempted multiple synthetic schemes and fails to be effectively synthesized out such micellar material, was carrying out scheme optimization
Afterwards, inventor finds a kind of route of synthesis finally, efficiently synthesizes out above-mentioned micellar material.Described synthetic schemes is as follows:
1) hyaluronate sodium (0.00882g) is dissolved in suitable quantity of water, obtains solution a；
2) (5- carboxy pentyl)-triphenylphosphinebromide (0.02079g), dmap (0.00053g), edc (0.01290g) put
In 50ml round-bottomed flask, appropriate dmso is added all to dissolve to solid drugs, 60 DEG C of agitating heating 1h, to activate (5- carboxyl penta
Base)-triphenylphosphinebromide carboxyl；
3) above-mentioned dmso solution is added dropwise over to solution a, and is added to appropriate tetrabutyl ammonium bromide, under room temperature
Stirring reaction 12h；
4) it is placed in the bag filter tying up with disposable dropper taking-up reaction solution and (take the bag filter of suitable length, be placed in
Soak in deionized water, cut the fine rule of two ends suitable length, be equally placed in deionized water, bag filter bubble after certain time
Soft, rubbed with the hands open, to facilitate addition solution later.Bag filter one, then the bag filter having more is turned back around several circles with fine rule,
With fine rule further around several circles, fasten, reactant liquor is transferred to wherein, and suitably rinsed several times with dmso, then according to same side
The other end that method ties up bag filter can complete), dialysis procedure will be tried one's best changes water more, and reaction solution is dialysed to the no taste of dmso
Road；
5) in the reactant liquor tiling culture dish dialysed, thickness about 3-5 millimeter, then empty with preservative film sealing, bundle, ice
Pre-freeze 12h under the conditions of -20 DEG C in case, freeze dryer lyophilization, obtain white solid and be this micellar material.
2. the sign of the micellar material synthesized by
Hyaluronic acid is dissolved in d respectively with the micellar material obtaining2O, uses h1- nmr ties to compound experiment products therefrom
Structure is verified, collection of illustrative plates is respectively as illustrated in fig. 1 and 2.Compared to Figure 1, new peak in 2.7 and 7.5 about in Fig. 2, through analyzing, 2.7
Left and right occur for methylene peak, and 7.5 fragrant area occur for phenyl ring peak. hyaluronate sodium described above and (5- carboxylic
Base amyl group)-triphenylphosphinebromide carried out esterification and generated comprising hyaluronic acid in structure and comprising (5- carboxyl penta again
Base)-triphenylphosphinebromide micellar material.
3. the preparation of curcumin micellar preparation and analysis
The foundation of 3.1 analysis methods and checking
3.1.1 liquid-phase chromatographic analysis
Liquid phase chromatogram condition is: chromatographic column: inertsil ods-sp (4.6 × 250mm, 5 μm)；Mobile phase: 0.5% vinegar
Aqueous acid-acetonitrile (40；60)；Flow velocity: 1ml/min；Detection wavelength: 425nm；Sample size: 20 μ l；Column temperature: 25 DEG C.
The drafting of standard curve: precision weighs curcumin 1 in 10ml brown volumetric flask, is rocked with analyzing pure methanol constant volume
Uniformly, it is mother solution.Take 1ml mother solution in 10ml brown volumetric flask, methanol constant volume shakes up.Take respectively 0.1ml, 0.5ml, 1ml, 5ml,
10ml diluent obtains the sample of 0.1 μ g/ml, 0.5 μ g/ml, 1 μ g/ml, 5 μ g/kg, 10 μ g/kg in 10ml volumetric flask constant volume, uses
Efficient liquid phase measures peak area.
Gained peak area, with shown in concentration records table 1, is depicted as standard curve (Fig. 3).Result shows, curcumin is in 0.1-
In 10 μ g/ml concentration ranges, linear relationship is good.
3.1.2 Particle Size Determination Method
The micelle preparing syringe was drawn 0.45 μm of microporous filter membrane in cillin bottle, took in right amount in particle size analyzer
Detection size, record data, preserve measurement data figure.
3.1.3 the assay method of envelop rate
Curcumin micellar solution after precision measured film respectively and curcumin micellar solution 1ml and the ep pipe of not crossing film
In, take 1ml methanol to be placed in ep pipe respectively with liquid-transfering gun, ultrasonic 5min, then all crosses microporous filter membrane, with high phase liquid phase measurement peak
Area, to cross film curcumin micellar solution peak area a1With not film curcumin micelle peak area a excessively2Ratio reaction micelle in wrap
The dose of envelope and the ratio of total dose, the calculating below equation of envelop rate (ee):
The preparation of 3.2 curcumin micellar preparations and condition optimizing
3.2.1 the optimization of solvent
Film dispersion method prepares blank micella: takes appropriate micellar material to add dehydrated alcohol, do not dissolve, a ultrasonic appropriate timing
Between dissolubility still bad, remove dehydrated alcohol with 40 DEG C of revolvings of Rotary Evaporators, add proper amount of acetone, dissolving, steamed with rotation
Send out 40 DEG C of revolvings removing acetone of instrument and transparent medicine film skeleton must be dried, add appropriate WAHAHA water, ultrasonic aquation 5min, syringe is inhaled
Take become solution, and with 0.45 μm of filtering with microporous membrane, obtain bright micellar material blank micella solution, 1500r/min from
Heart 5min removes impurity, records particle diameter with particle size analyzer.Particle diameter distribution statistical result as shown in figure 4, the concentration class of particle diameter distribution relatively
Low.
Inventor is optimized to solvent, and dehydrated alcohol is made into acetone, prepares curcumin micelle using following methods
Preparation: take micellar material (0.01007g) and curcumin (0.00051g) to add acetone 10ml, dissolving, 40 DEG C of rotations of Rotary Evaporators
Turn evaporation, remove acetone, form transparent medicine film skeleton, be placed in 80 DEG C of oil baths, after several minutes, add about 40 DEG C of WAHAHA pure
Water, after reaction certain time, takes out and is cleaned by ultrasonic ultrasonic aquation 5min in instrument, obtains solution syringe and draws, crosses 0.45 μ
M microporous filter membrane, records particle diameter with particle size analyzer.Particle diameter distribution statistical result is as shown in figure 5, the concentration class of particle diameter distribution is very high.
3.2.2 the optimization of ultrasonic time
Precision weighs 15 micellar material and three parts of 0.5 curcumin, in 50ml eggplant type bottle, takes 10ml acetone molten
Solution, ultrasonic several minutes make it dissolve, and revolving removes acetone, are placed in 80 DEG C of oil baths, add about 40 DEG C of WAHAHA water, and add little
Magneton, so as to rotation solidification, takes out ultrasonic aquation 5min, 10min, 15min in ultrasonic cleaning instrument after several minutes.Cross
0.22 μm of microporous filter membrane, obtain yellow homogeneous curcumin micellar solution.Measure particle diameter and envelop rate respectively, with particle diameter and
Envelop rate is inspection target, investigates the different impacts to curcumin micelle particle diameter and envelop rate for the ultrasonic time.Result such as table 2 and
Shown in Fig. 6-8, during ultrasonic 15min, mean diameter is minimum, particle diameter distribution is concentrated most, and envelop rate is maximum.
The different impact to micelle mean diameter and envelop rate for the ultrasonic time of table 2
3.2.3 the ratio optimization of micellar material and curcumin
Precision weighs 15 micellar material and 0.5 curcumin, 20 micellar material and 0.5mg curcumin respectively, and 25
Micellar material and 0.5 curcumin, in 50ml eggplant type bottle, take 10ml acetone solution, ultrasonic several minutes make it dissolve, rotation
Acetone is evaporated off, is placed in 80 DEG C of oil baths, add about 40 DEG C of WAHAHA water, and add little magneton so as to rotation solidifies, several minutes
After take out in be cleaned by ultrasonic instrument in ultrasonic aquation 5min.Cross 0.45 μm of microporous filter membrane, obtain yellow homogeneous and bright Rhizoma Zingiberis Recens
Flavin micellar solution.Measure particle diameter and envelop rate respectively, and be inspection target particle diameter and envelop rate, investigate different micelle materials
Material and the impact to curcumin micelle particle diameter and envelop rate for the curcumin ratio.Result as shown in table 3 and Fig. 9-11, micellar material
It is that during 50:1, particle diameter is minimum with curcumin ratio, and micellar material and curcumin ratio are envelop rate during 30:1 from the point of view of envelop rate
Highest.Micellar material and curcumin ratio are that when during 30:1, particle diameter is likely due to the reason excessive measure, absorption cell is not dry
Only, have last time measure only 0.45 μm of microporous filter membrane when debris, so particle diameter is larger.Therefore consider, ratio of greater inequality
It is micellar material and curcumin ratio selection 30:1.
Table 3 micellar material and the impact to micelle mean diameter and envelop rate for the curcumin
3.2.4 the optimization of solidification temperature
Precision weighs 15 micellar material and three parts of 0.5 curcumin, in 50ml eggplant type bottle, takes 10ml acetone molten
Solution, ultrasonic several minutes make it dissolve, and revolving removes acetone, are respectively placed in 40 DEG C, 60 DEG C, 80 DEG C of oil baths, add about 40 DEG C of baby
Heartily water, and add little magneton so as to rotation solidification, take out ultrasonic aquation 5min in ultrasonic cleaning instrument after several minutes.Cross
0.22 μm of microporous filter membrane, obtains the homogeneous and bright curcumin micellar solution of yellow.Measure particle diameter and envelop rate respectively, and
It is inspection target particle diameter and envelop rate, investigate the different impacts to curcumin micelle particle diameter and envelop rate for the solidification temperature.Result
As shown in table 5 and Figure 12-14, from the point of view of particle diameter, solidification temperature selects particle diameter when 40 degrees Celsius minimum, from the point of view of envelop rate, Gu
Change temperature and select 80 DEG C preferably, therefore, consider, optimum solidification temperature is 80 DEG C.
The different impact to mean diameter and envelop rate for the solidification temperature of table 5
3.2.5 curcumin micellar preparation optimum results
By single factor exploration, substantially determine optimum prescription and the preparation technology of common curcumin micellar preparation: accurate
Weigh 15 micellar material and 0.5 curcumin, be placed in 50ml eggplant type bottle, take 10ml acetone to be placed in eggplant type bottle, gently shake
Swing, ultrasonic several minutes make it dissolve, revolving removes acetone until forming semi-transparent jonquilleous film on a rotary evaporator, is placed in 80
DEG C heat-collecting magnetic stirring device, controls temperature to be 80 DEG C, and stirs, take about 10ml WAHAHA water warm in a water bath with beaker simultaneously
Heat.After heating certain time, it is added dropwise over about 40 DEG C of WAHAHA water, and add little magneton so as to rotation solidifies, after several minutes
Take out ultrasonic aquation 5min in ultrasonic cleaning instrument.Solution crosses 0.45 μm of microporous filter membrane, you can obtain yellow homogeneous and bright
Curcumin micellar solution.
3.2.6 orthogonal design optimization formulation is tested
Single factor exploration experiment finds the ratio of micellar material and curcumin, ultrasonic time, three factors of solidification temperature to reality
Test impact all larger, therefore these three factors investigated further by orthogonal experimental design method, and using particle diameter and envelop rate as
Final evaluation index, carries out Three factors-levels orthonormal design of experiments, and empirical factor and level are shown in Table 6, and experimental result is shown in Table 7.
Table 6 curcumin micellar preparation orthonormal design of experiments factor and level
Table 7 orthonormal design of experiments table l9 (34) and experimental result
According to orthogonal test, during using particle diameter as reference index, extreme difference reflects the ratio shadow of micellar material and curcumin
Ring and be more than solidification temperature, more than ultrasonic time, each factor optimal level is: micellar material is 30:1 with the ratio of curcumin；Ultrasonic
Time is 15min；Solidification temperature is 80 DEG C.During using envelop rate as reference index, the impact of solidification temperature is more than micellar material
With the ratio of curcumin, more than ultrasonic time, each factor optimal level is: the ratio of micellar material and curcumin: 50:1；Ultrasonic time
For 15min；Solidification temperature is 80 DEG C., all in below 400nm, difference is little each other, institute for this test gained micelle particle diameter
So that by envelop rate, as Primary Reference index, therefore the optimum prescription of test and optimised process are: the ratio of micellar material and curcumin:
50:1；Ultrasonic time is 15min；Solidification temperature is 80 DEG C.
The foregoing is only presently preferred embodiments of the present invention, not in order to limit the present invention, all spirit in the present invention and
Within principle, any modification, equivalent substitution and improvement made etc., should be included within the scope of the present invention.
Claims (10)
1. a kind of Mitochondrially targeted micellar material is it is characterised in that by a hydroxyl on oligomerization hyaluronic acid and (5- carboxyl penta
Base) the 5- carboxyl of-triphenylphosphinebromide is formed by connecting by ester bond, and its formula is:
Wherein, n is positive integer, and is the molecular weight of described oligomerization hyaluronic acid less than 10000da.
2. a kind of preparation method of Mitochondrially targeted as claimed in claim 1 micellar material is it is characterised in that by making hyalomitome
Sour sodium and (5- carboxy pentyl)-triphenylphosphinebromide react under the catalytic action of dmap, edc and obtain described Mitochondrially targeted glue
Beam material.
3. preparation method according to claim 2 is it is characterised in that comprise the following steps:
1) prepare the aqueous solution of hyaluronate sodium；
2) dmso solution will in (5- carboxy pentyl)-triphenylphosphinebromide, dmap, edc solution dmso, be prepared into, and in 60 DEG C
Lower stirring 1 hour；
3) described dmso solution is added dropwise in the aqueous solution of described hyaluronate sodium, and is added thereto to tetrabutyl phosphonium bromide
Ammonium, stirs at room temperature and terminates to reaction, the weight of the hyaluronate sodium being added and (5- carboxy pentyl)-triphenylphosphinebromide
Than for 0.00882:0.02079；
4) by step 3) reaction product solution that obtains is transferred to bag filter, dialyses in deionized water, to removing dms0；
5) the reaction product solution lyophilization through dialysis, that is, obtain described Mitochondrially targeted micellar material.
4. a kind of Mitochondrially targeted curcumin micellar preparation is it is characterised in that wrapped up by the micellar material described in claim 1
Curcumin forms.
5. a kind of preparation method of curcumin micellar preparation Mitochondrially targeted as claimed in claim 4 is it is characterised in that include
Following steps:
1) micellar material described in claim 1 and curcumin are dissolved in equipped with the container of solvent；
2) rotary evaporation removes described solvent, forms medicine film skeleton to described container inner wall；
3) described container is placed in oil bath, and adds water in described container, so that medicine film is solidified；
4) ultrasonic aquation；
5) filtering with microporous membrane, obtains described curcumin micellar preparation.
6. preparation method according to claim 5 is it is characterised in that the weight ratio of described micellar material and described curcumin
For 30:1-50:1.
7. preparation method according to claim 5 is it is characterised in that described solvent is acetone.
8. preparation method according to claim 5 is it is characterised in that step 3) in solidify the temperature of described medicine film be 40-
80℃.
9. preparation method according to claim 5 is it is characterised in that step 4) in time of ultrasonic aquation be 5-15min.
10. the preparation method according to any one of claim 5-9 it is characterised in that described preparation method include following
Step:
1) the described micellar material than 30:1 for the weight and curcumin are dissolved in acetone and make acetone soln, load in container；
2) acetone described in rotary evaporation, forms medicine film skeleton to described container inner wall；
3) described container is placed in 80 DEG C of oil baths, is added dropwise over about 40 DEG C of water, and add little magneton stirring, make medicine film solid
Change；
4) ultrasonic aquation 15min；
5) 0.45 μm of filtering with microporous membrane, obtains described curcumin micellar preparation.
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