CN109125739A - Multifunctional macromolecule Micellar drug delivery system and its preparation method and application - Google Patents
Multifunctional macromolecule Micellar drug delivery system and its preparation method and application Download PDFInfo
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Abstract
The present invention designs targeted delivery of drugs technical field, it is related to multifunctional macromolecule Micellar drug delivery system and its preparation method and application, by polyethylene glycol lysine block copolymer with 3,3 '-two thiobis the third imido dimethyl phthalate dihydrochlorides are chemically modified partial side-chain and obtain the group with sulfydryl, then react the ten diborane disodium salt of boron neutron capture therapy drug sulfydryl with sulfydryl with side chain by disulfide bond and act on forming stable multifunctional macromolecule micella by the free disulfide bond crosslinking of self assembly and part in aqueous solution；Prepared macromolecule micelle can deliver ten diborane disodium of drug sulfydryl or other drugs or prodrug for containing sulfydryl containing sulfydryl, increase the medicament contg delivered into tumour cell, improve the targeted therapy effect to tumour.
Description
Technical field
The present invention designs targeted delivery of drugs technical field, is related to multifunctional macromolecule Micellar drug delivery system and its system
Preparation Method and application have the stability that drug delivery system is improved by disulfide bond crosslinking, and endosome of escaping is to tumour cell
The interior new opplication for realizing high potency drugs delivering and response glutathione drug release, relates particularly to and radiotherapy and/neutron absorption
The application for the treatment of/or chemotherapeutics in preparation treatment tumour or malignant tumour.
Background technique
Drug delivery system is a kind of mode of a kind of drug-loading system and pharmaceutical preparation, mainstay object carrier material
(such as high molecular material) is used to load bioactive substance/drug to construct drug delivery system, and delivers it to lesion portion
Position plays drug effect.Drug delivery system has the toxic side effect for reducing drug, extends blood circulation time and keeps having treatment
Drug targeting is delivered to lesions position by the blood concentration of effect, and the control release of drug and reduction drug are in normal tissue
The significances such as exposure.Therefore, the research and development of drug delivery system are considered as the basic fundamental system of drug therapy in future, by state
Inside and outside extensive concern has become medicine, chemistry, biomaterial, the research hotspot in the multi-crossed disciplines such as macromolecule field.It is high
Molecule micella is one kind of drug delivery system, is mainly loaded drug by being self-assembly of by amphipathic nature block polymer
In core, the package or solubilising to drug are completed, outer layer is generally made of polyethylene glycol polymer, can also be by macromolecule
End or side chain graft hydrophobic molecule or sulfydryl in karyomorphism at crosslinking, to increase the stability of macromolecule micelle.High score
Sub- micella has size tunable, and drugloading rate is big, prepares the advantages that facilitating, and is a kind of novel drug delivery system.
Malignant tumour, and i.e. cancer, are to lead to one of Important cause of disease of human death, and in China, malignant tumour comes institute
There is first of disease death reason, only the neopathy number of cases of malignant tumour in 2014 is more than 3,800,000, average daily more than 1
Ten thousand people are diagnosed as cancer, have 7 people to be diagnosed as cancer per minute.Malignant tumour has become the important of Chinese national health
The tumour status of killer, China have, and disease incidence is high, the features such as death rate height.Malignant tumour has complexity, at present for swollen
Early diagnosis, the treatment of the comprehensive means such as radiation and chemotherapy are mainly taken in the treatment of tumor.It is maximum to lack using above-mentioned treatment method
It is trapped in tumour cell and normal cell, without selectivity, the side effect caused by normal cell is too big.Therefore, research and development have
The drug delivery system of cancer target has significant application value.
In the radiotherapy of tumour, boron neutron capture therapy is a kind of selective cancer radiation therapy mode, former
Reason be allow tumour cell intake boracic (10B drug), is radiated by thermal beam, boron atom (10B thermal neutron) is captured, because10B have big thermal-neutron reaction section, can effectively capture thermal neutron, then occur nuclear fission reaction, generate high-energy Ah
Your method particle (α) and7Li core destroys cancer cell by the α particle of generation, thorough to the irreversible fatal injury of tumour cell
Its genetic structure is destroyed at bottom, prevents it from repairing and apoptosis, achievees the purpose that oncotherapy, and hanker sub-line and relatively pacify
Entirely, to no intake boron (10B the normal cell etc. of drug) does not generate harmful effect.However, what boron neutron capture therapy generated
The range of α particle is extremely short, about 5-9 microns, connects cytoproximal diameter, and the ionization radiation injury effect that these particles generate is only limitted to
Intake10The cell of B, therefore, in order to reach effectively kill cancer cell therapeutic effect, need by boracic (10B drug) is passed
It is sent in tumour cell, therapeutic effect could be obtained.Other radiation treatment modes use radioactive isotope, and tumour boron
The boron-containing compound of neutron capture therapy does not have on-radiation, safer.Boron neutron capture therapy at home and abroad, such as day
This, the U.S. and China successfully cures several patients, in particular for the tumour for the excision that is difficult to perform the operation, such as melanoma, breast cancer,
Brain tumor etc., achieves success.So far, there are two types of boracic (10B drug), ten diborane disodium salt (sodium of sulfydryl
Borocaptate, BSH) and p- boron phenylalanine (boronophenylalanine, BPA).Ten diborane disodium salt of sulfydryl is
Boron cluster class compound has good hydrolysis and metabolic stability under biotic environment, and boron content is high, in identical dosage water
Under flat compared with p- boron phenylalanine, can carry more boron (10It B), is current most effective boron neutron capture therapy drug
One of, but due to properties such as its electronegativity, lack tumour cell/tissue target function, seldom by tumour cell endocytosis,
It is difficult to reach the effective of boron neutron capture therapy in tumour cell10B concentration influences the boron neutron capture therapy of tumour, structure
Formula is as follows:
Therefore, increase targeting and intracellular levels of drugs of ten this drug of diborane disodium salt of sulfydryl to tumour cell,
It is of great significance, ten diborane disodium salt of sulfydryl can be delivered to tumor tissues and cell by building drug delivery system, be increased
Add in tumour cell10B concentration improves the therapeutic effect to tumour.Currently, ten diborane disodium salt of sulfydryl imitates the treatment of tumour
Fruit need to be improved, and also lacks the multifunctional macromolecule micelle nano drug delivery to ten diborane disodium salt high-efficiency delivery of sulfydryl
System, to improve tumour cell to the endocytosis of ten diborane disodium salt of sulfydryl, the targeted delivery and boron neutron capture therapy of drug
Effect.
Summary of the invention
The object of the present invention is to provide a kind of multifunctional macromolecule Micellar drug delivery system and its preparation method and application,
Specific technical solution are as follows:
Multifunctional macromolecule Micellar drug delivery system, for modified polyethylene glycol lysine block copolymer and band
The boron neutron capture therapy drug of sulfydryl passes through the free disulfide bond crosslinking of self assembly and part in aqueous solution and acts on the height to be formed
Molecule micella.
The polyethylene glycol lysine block copolymer of the modification, with 3,3 '-two thiobis the third imido dimethyl phthalates
Dihydrochloride is chemically modified polyethylene glycol lysine block copolymer partial side-chain and obtains the group with sulfydryl.
The boron neutron capture therapy drug with sulfydryl is drug molecule or prodrug containing sulfydryl；
The boron neutron capture therapy drug with sulfydryl is ten diborane disodium salt of sulfydryl.
Chemical reaction process are as follows:
Wherein the block copolymer synthesis process of ten diborane disodium salt of sulfydryl crosslinking is；
Wherein, m is the number of repeat unit of polyethylene glycol, in the range of 3-500；
N is the unit number of polycation, in the range of 3-300；
X is the number of repeat unit of polylysine, in the range of 1-100；
Y is that 3,3 '-two thiobis the third imido dimethyl phthalate dihydrochlorides carry out chemistry to polyethylene glycol lysine side-chain
The number of repeat unit of ten diborane disodium salt of drug molecule sulfydryl is connected with after modification by disulfide bond, in the range of 1-100；
Z is that 3,3 '-two thiobis the third imido dimethyl phthalate dihydrochlorides carry out chemistry to polyethylene glycol lysine side-chain
The number of repeat unit of modification, in the range of 1-100.
The multifunctional macromolecule Micellar drug delivery system is substantially former with polyethylene glycol lysine block copolymer
Material, the compounds such as 3,3 '-two thiobis the third imido dimethyl phthalate dihydrochlorides generate sulfydryl in side chain, and it is total to form polymer blocks
Polymers, the high-molecular block copolymer pass through the drug molecule containing sulfydryl such as sulfydryl and ten diborane disodium salt of sulfydryl or preceding
Medicine reaction, finally by macromolecule micelle is self-assembly of, unreacted free sulfhydryl group can be increased high by disulfide bond crosslinking
The stability of molecule micella.
Drug containing sulfydryl in conjunction with high-molecular block copolymer, there is polycation by disulfide bond in macromolecule micelle
Segment and free sulfydryl are used for the core of cross-linked polymer micella.The particle size of the macromolecule micelle eventually formed is in 10-200
Between nanometer.
The preparation method of the multifunctional macromolecule Micellar drug delivery system, comprising the following steps:
(1) synthesis of high molecular material
Polyethylene glycol lysine block copolymer is dissolved in borate buffer, while 3,3 '-two thiobis third are sub-
Propylhomoserin dimethyl ester dihydrochloride is dissolved in borate buffer, two kinds of solution hybrid reaction 45 minutes at room temperature, then will reaction
Product is dialysed three times in the phosphate of pH7.4, and dithiothreitol (DTT) reaction is then added, is dialysed with phosphate buffer solution and pure water
Product obtains modified polyethylene glycol lysine block copolymer macromolecule material three times, finally by freeze-drying；
(2) crosslinking of drug and high molecular material prepares macromolecule micelle
Modified polyethylene glycol lysine block copolymer is dissolved in HEPES buffer solution, by ten diborane two of sulfydryl
Sodium is dissolved in HEPES buffer solution, then mixes two kinds of solution, is stirred to react, then product is used to HEPES buffer solution respectively
It dialyses with pure water, carries the molten macromolecule micelle of ten diborane disodium of medicine sulfydryl by being self-assembly of.
Polyethylene glycol lysine block copolymer used in ingredient and 3,3 '-two the third imidic acids of thiobis in step (1)
The molar ratio of dimethyl ester dihydrochloride is 1:1, but is not limited only to 1:1.3,3 '-two thiobis used in ingredient in step (2)
The polyethylene glycol lysine block copolymer and ten diborane disodium of sulfydryl of third imido dimethyl phthalate dihydrochloride modified side chain
Molar ratio be 1:1, but the ingredient proportion of ten diborane disodium of sulfydryl can be increased.
Multifunctional macromolecule Micellar drug delivery system prepared by the present invention by disulfide bond and contains the drug of sulfydryl
Or prodrug connection, the glutathione by responding internal high concentration realize the control release function of drug, while in macromolecule
Amino cation can gulp down in the cell during protonation come realize escape endosome or lysosome function, decline
The part free sulfhydryl group of free polymer side chain can enhance macromolecule micelle in normal physiological conditions by being crosslinked micella core
Under stability, and the disulfide bond in tumour cell can by respond glutathione disconnect, realize it is crosslinked polymeric
Dissociation.Therefore, which can be in the application in drug delivery, specifically and radiotherapy
With/the application of neutron capture therapy/or chemotherapeutics in preparation treatment tumour or malignant tumour.The multifunctional macromolecule micella
Drug delivery system is the drug delivery system with tumor-targeting as a kind of pharmaceutical carrier；The macromolecule micelle can be with
Radiotherapeutic drug or chemotherapeutics or tumor suppressor gene and nucleic acid etc. are delivered, drug or gene orientation are delivered to lesions position,
It is a kind of macromolecule micelle of cancer target.
The invention has the following advantages:
1, the type for increasing macromolecule micelle drug delivery system has widened the application field of macromolecule micelle, favorably
In the research and development of drug delivery system and new medicinal preparation.
2, multifunctional macromolecule Micellar drug delivery system of the present invention, high concentration in tumour cell can be responded by having
Drug controlled release and micella dissociation may be implemented for being crosslinked micella core and being crosslinked with drug in the disulfide bond of glutathione response
Function；Polycation in macromolecule micelle has the function that lysosome and endosome are destroyed in tumour cell endocytic processes,
It realizes the purpose that into tumour cell high potency drugs deliver, it is difficult and to tumour cell to solve existing drug targeting tumour cell
The problems such as interior drug delivery is insufficient.
3, pharmaceutical preparation is novel, and drug delivery effect is good, can be with targets neoplastic cells.
4, preparation method is simple, and required equipment is conventional equipment.
Detailed description of the invention
Fig. 1 is the grain size distribution that embodiment 1 characterizes macromolecule micelle；
Fig. 2 is the transmission electron microscope picture that embodiment 1 characterizes macromolecule micelle pattern；
Fig. 3 is the drug release result that embodiment 4 characterizes macromolecule micelle response glutathione and dithiothreitol (DTT)；
The tumour cell that Fig. 4 a is embodiment 4 to be tested with plasma mass is to ten diborane disodium of sulfydryl and carries medicine high score
The comparison result of the opposite breast cancer cell endocytosis amount of sub- micella；
The tumour cell that Fig. 4 b is embodiment 4 to be tested with plasma mass is to ten diborane disodium of sulfydryl and carries medicine high score
The comparison result of the relative melanin oncocyte endocytosis amount of sub- micella；
Fig. 5 a is that the ten diborane disodium of sulfydryl of embodiment 4 and medicine-carrying polymer micella are used for in the boron of breast cancer cell
Neutron capture therapy result figure；
Fig. 5 b is the ten diborane disodium of sulfydryl of embodiment 4 and medicine-carrying polymer micella for the boron to melanoma cells
Neutron capture therapy result figure.
Specific embodiment
The present invention is described further below in conjunction with several embodiments.It should absolutely prove, following embodiments are only used
In illustrating the present invention, but do not limit the invention in any way.
Embodiment 1:
The present embodiment is with polyethylene glycol lysine block copolymer and 3,3 '-two the third imido of thiobis dimethyl phthalates two
Hydrochloride is material construction macromolecule micelle medicament carrier system, and drug molecule used is ten diborane disodium salt of sulfydryl, technique
Steps are as follows:
(1) synthesis of high molecular material
The polyethylene glycol of 3,3 '-two thiobis the third imido dimethyl phthalate dihydrochloride modified side chains of 50mg is relied into ammonia
Sour block copolymer is dissolved in the HEPES buffer solution (10mM, pH7.4) of 15ml, and the medicinal ten diborane disodium of sulfydryl of 34mg is molten
In the HEPES buffer solution (10mM, pH7.4) of 4ml, then two kinds of solution are mixed, are stirred to react 6 hours, then by product
It is dialysed 24 hours with HEPES buffer solution and pure water respectively, carries the molten height of ten diborane disodium of medicine sulfydryl by being self-assembly of
Molecule micella.
(2) preparation of the crosslinking of drug and high molecular material and macromolecule micelle
The polyethylene glycol of 3,3 '-two thiobis the third imido dimethyl phthalate dihydrochloride modified side chains of 100mg is relied into ammonia
Sour block copolymer is dissolved in the HEPES buffer solution (10mM, pH7.4) of 10ml, and the medicinal ten diborane disodium of sulfydryl of 75mg is molten
In the HEPES buffer solution (10mM, pH7.4) of 10ml, then two kinds of solution are mixed, are stirred to react 1 hour, then by product
It is dialysed 24 hours with HEPES buffer solution and pure water respectively, carries the molten height of ten diborane disodium of medicine sulfydryl by being self-assembly of
Molecule micella.
The particle size of its macromolecule micelle prepared is between 10-200nm, as shown in Figure 1, resulting macromolecule micelle
There is the uniform feature of particle diameter distribution by projection Electronic Speculum observation, as shown in Figure 2.
Embodiment 2:
The present embodiment is with polyethylene glycol lysine block copolymer and 3,3 '-two the third imido of thiobis dimethyl phthalates two
Hydrochloride is material construction macromolecule micelle medicament carrier system, and drug molecule used is ten diborane disodium salt of sulfydryl, technique
Steps are as follows:
(1) synthesis of high molecular material
Under conditions of 25 DEG C, the polyethylene glycol lysine block copolymer of 100mg, which is dissolved in 10mL concentration, is
100mM, in the borate buffer that pH value is 9.0, while by the 3 of 50mg, 3 '-two thiobis the third imido dimethyl phthalate dihydrochlorides
Being dissolved in 12mL concentration is 100mM, in the borate buffer that pH value is 9.0, two kinds of solution hybrid reaction 60 minutes at room temperature, so
Reaction product is dialysed three times in 10mM, the phosphate of pH7.4 afterwards, 70mg dithiothreitol (DTT) is then added and reacts 50 minutes,
Then three times with phosphate buffer solution and pure water dialysis, 3,3 '-two the third imidic acids of thiobis two are obtained finally by freeze-drying
The polyethylene glycol lysine block copolymer of methyl esters dihydrochloride modified side chain.
(2) preparation of the crosslinking of drug and high molecular material and macromolecule micelle
By the polyethylene glycol lysine of 3,3 '-two thiobis the third imido dimethyl phthalate dihydrochloride modified side chains of 5mg
Block copolymer is dissolved in the HEPES buffer solution (10mM, pH7.4) of 1ml, and sulfhydryl modified DNA plasmid is dissolved in 0.1ml's
HEPES buffer solution (10mM, pH7.4) then mixes two kinds of solution, is vigorously stirred reaction 10 minutes, 50 μ L are then added
Dimethyl sulfoxide stands 3-10 hours, and then product is put in ultra-filtration centrifuge tube and is washed with pure water or HEPES buffer solution
It purifies 3-5 times and goes out decontamination, finally obtain the macromolecule micelle of package DNA plasmid.
Embodiment 3:
The present embodiment is with polyethylene glycol lysine block copolymer and 3,3 '-two the third imido of thiobis dimethyl phthalates two
Hydrochloride is material construction macromolecule micelle medicament carrier system, and drug molecule used is ten diborane disodium salt of sulfydryl, technique
Steps are as follows:
(1) synthesis of high molecular material
Under conditions of 25 DEG C, the polyethylene glycol lysine block copolymer of 100mg, which is dissolved in 10mL concentration, is
100mM, in the borate buffer that pH value is 9.0, while by the 3 of 150mg, 3 '-two thiobis the third imido dimethyl phthalate disalts acid
It is 100mM that salt, which is dissolved in 15mL concentration, and pH value is in 9.0 borate buffer, two kinds of solution at room temperature divides by hybrid reaction 120
Then clock dialyses reaction product three times in 10mM, the phosphate of pH7.4,200mg dithiothreitol (DTT) reaction 60 is then added
Minute, then three times with phosphate buffer solution and pure water dialysis, 3,3 '-two the third imido of thiobis are obtained finally by freeze-drying
The polyethylene glycol lysine block copolymer of dimethyl phthalate dihydrochloride modified side chain.
(2) preparation of the crosslinking of drug and high molecular material and macromolecule micelle
By the polyethylene glycol lysine of 3,3 '-two thiobis the third imido dimethyl phthalate dihydrochloride modified side chains of 5mg
Block copolymer is dissolved in the HEPES buffer solution (10mM, pH7.4) of 2ml, and 2mg sulfydryl camptothecine is dissolved in the dimethyl of 200 μ L
In sulfoxide, then Polymer Solution is placed in ultrasound, sulfydryl camplotheca acuminata aqueous slkali is rapidly joined, continuous ultrasound 5 minutes, then
The macromolecule micelle of preparation is stood 3-10 hours, then product is put in ultra-filtration centrifuge tube with pure water or HEPES buffering
Solution washing purifies 3-5 times and goes out decontamination, finally obtains the macromolecule micelle of package DNA plasmid.
Embodiment 4:
Application of the multifunctional macromolecule micella in tumour boron neutron capture therapy is loaded with medicine for what is prepared in embodiment 1
The macromolecule micelle of ten diborane disodium of object sulfydryl is configured to aqueous solution or normal saline solution, is then arrived by intravenous injection
With tumor-bearing mice, injection dosage 100mg/kg after injection drug 24 hours, is irradiated tumor locus 1 hour with sub-line is hankered,
The dosage for hankering sub-line is (1.6-2.2 × 1012neutron/cm2), a tumor size is then measured every three days, is realized swollen
The boron neutron capture therapy of tumor.
The glutathione or identical dense of the present invention that medicine multifunctional macromolecule micella is carried under the conditions of inside tumor cells
The dithiothreitol (DTT) of degree is horizontal (2-10mM), the disulfide bond being connected between drug and macromolecule can be quickly cut off, by drug
It is released from macromolecule micelle, and under normal physiological conditions, glutathione content level is lower (0.01mM), drug base
It is not discharged in sheet, as shown in figure 3, in this way, may be implemented to drug molecule, such as the tumour of ten diborane disodium of sulfydryl
The control of cell-targeting discharges.Multifunctional macromolecule micella of the present invention may be implemented to drug or prodrug containing sulfydryl
Response tumour cell glutathion inside concentration control release.
Load medicine multifunctional macromolecule micella of the present invention can deliver ten diborane disodium of sulfydryl to tumour cell, use
Macromolecule micelle acts on tumour cell with exclusive use ten diborane disodium of sulfydryl and compares, and can significantly improve in tumour cell
Medicament contg, for example with macromolecule micelle and ten diborane disodium of sulfydryl act on 4T1 breast cancer cell core B16F10 simultaneously
After melanoma cells 24 hours, the content of drug is 10 times of control group or more in the tumour cell of macromolecule micelle effect,
As shown in figures 4 a and 4b.Multifunctional macromolecule micella of the present invention have by after tumour cell endocytosis escape lysosome,
The Functional Design of endosome can deliver more drugs into cell, realize the high potency drugs delivering of targets neoplastic cells.
Load medicine multifunctional macromolecule micella of the present invention can effectively kill cancer cell by boron neutron capture therapy,
Therapeutic effect far superior to uses ten diborane disodium of sulfydryl, such as by ten diborane disodium of medicine-carrying polymer micella and sulfydryl
(wherein the content of drug is all 10 μM) is respectively acting on 4T1 breast cancer cell core B16F10 after melanoma cells 24 hours,
With sub-line irradiating cell is hankered 1 hour, then cultivates 24 hours and go test cell survival rate, as the result is shown medicine-carrying polymer micella
Most of tumour cell can be killed, and ten diborane disodium of sulfydryl does not almost have lethal effect to cell, such as Fig. 5 a and Fig. 5 b
It is shown.Medicine-carrying polymer micella of the present invention can effectively realize the boron neutron capture therapy to tumour.
Claims (7)
1. multifunctional macromolecule Micellar drug delivery system, which is characterized in that total for modified polyethylene glycol lysine block
Polymers and the therapeutic agent with sulfydryl pass through the free disulfide bond crosslinking of self assembly and part in aqueous solution and act on the high score to be formed
Sub- micella.
2. multifunctional macromolecule Micellar drug delivery system according to claim 1, which is characterized in that the modification
Polyethylene glycol lysine block copolymer, with 3,3 '-two thiobis the third imido dimethyl phthalate dihydrochlorides to polyethylene glycol-
Polylysine block copolymer portion side chain, which is chemically modified, obtains the group with sulfydryl.
3. multifunctional macromolecule Micellar drug delivery system according to claim 1, which is characterized in that the band sulfydryl
Therapeutic agent be drug molecule or prodrug containing sulfydryl.
4. multifunctional macromolecule Micellar drug delivery system according to claim 3, which is characterized in that the band sulfydryl
Therapeutic agent be ten diborane disodium salt of sulfydryl.
5. multifunctional macromolecule Micellar drug delivery system according to claim 4, which is characterized in that ten diborane of sulfydryl
Disodium salt crosslinking block copolymer synthesis process be；
Wherein, m is the number of repeat unit of polyethylene glycol, in the range of 3-500；
N is the unit number of polycation, in the range of 3-300；
X is the number of repeat unit of polylysine, in the range of 1-100；
Y is that 3,3 '-two thiobis the third imido dimethyl phthalate dihydrochlorides are chemically modified polyethylene glycol lysine side-chain
The number of repeat unit of ten diborane disodium salt of drug molecule sulfydryl is connected with by disulfide bond afterwards, in the range of 1-100；
Z is that 3,3 '-two thiobis the third imido dimethyl phthalate dihydrochlorides are chemically modified polyethylene glycol lysine side-chain
Number of repeat unit, in the range of 1-100.
6. special according to claim 1 to the preparation method of 5 described in any item multifunctional macromolecule Micellar drug delivery systems
Sign is, comprising the following steps:
(1) synthesis of high molecular material
Polyethylene glycol lysine block copolymer is dissolved in borate buffer, while by 3,3 '-two the third imidic acids of thiobis
Dimethyl ester dihydrochloride is dissolved in borate buffer, two kinds of solution hybrid reaction 45 minutes at room temperature, then by reaction product
It dialyses in the phosphate of pH7.4 three times, dithiothreitol (DTT) reaction is then added, with phosphate buffer solution and pure water dialysis product
Three times, modified polyethylene glycol lysine block copolymer macromolecule material is obtained finally by freeze-drying；
(2) crosslinking of drug and high molecular material prepares macromolecule micelle
Modified polyethylene glycol lysine block copolymer is dissolved in HEPES buffer solution, by the sulfydryl 12 as drug
Borine disodium is dissolved in HEPES buffer solution, then mixes two kinds of solution, is stirred to react, then delays product with HEPES respectively
Solution and pure water dialysis are rushed, carries the molten macromolecule micelle of ten diborane disodium of medicine sulfydryl by being self-assembly of.
7. according to claim 1 to 5 described in any item multifunctional macromolecule Micellar drug delivery systematic differences,
It is characterized in that, the multifunctional macromolecule Micellar drug delivery system is used to prepare the medicine in treatment tumour or malignant tumour
Object.
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