CN107115319A - A kind of biological safety carbon quantum dot load adriamycin complex and its preparation method and application - Google Patents
A kind of biological safety carbon quantum dot load adriamycin complex and its preparation method and application Download PDFInfo
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Abstract
Adriamycin complex is loaded the invention discloses a kind of biological safety carbon quantum dot, its preparation method includes：(1) it is 56 by volume by milk and deionized water:45 mixing, are subsequently placed in 160 200 DEG C of 2 4h of heating；(2) room temperature is cooled to, 30 45min are centrifuged under 10000 12000r/min, supernatant is taken；(3) membrane filtration, obtains carbon quantum dot solution；(4) the carbon quantum dot aqueous solution is prepared；(5) the adriamycin aqueous solution is prepared；(6) the carbon quantum dot aqueous solution, the adriamycin aqueous solution and PBS solution are mixed, pH=7.4 mixed solution is made；(7) lucifuge is reacted, and is dialysed, and is made.The preparation method is simple, and adriamycin load factor is high, and obtained complex biological is safe, and the killing-efficiency to tumour cell is significantly improved, the lethal obvious reduction to normal cell, available for tumor disease therapeutic.
Description
Technical field
The invention belongs to pharmaceutical technology field, and in particular to a kind of biological safety carbon quantum dot loads adriamycin complex
And its preparation method and application.
Background technology
Cancer is one of important diseases of current threat human life and health, has millions of people to die from cancer every year, on
The treatment of cancer is the topic that medical personnel and scientific research personnel constantly explore.In addition to operative treatment, chemotherapy is a kind of
Important treatment method.Chemotherapeutics species for different type cancer is various, and adriamycin is a kind of spectrum cancer therapy drug, right
Kinds of tumor cells has lethal effect, is clinically widely used in ALL, lymthoma, breast cancer, ovum
Nest cancer, lung cancer, stomach cancer, liver cancer, G. cephalantha etc..But because tumour cell is single to the intake approach of adriamycin, adriamycin
Enter core to play a role difficulty, therefore dosage can be only achieved the purpose of killing tumor cell enough, adriamycin can be killed simultaneously in addition
Hinder normal cell, cause concurrent many adverse reactions during medicine use.Therefore novel Drug Delivery Systems gradually cause researcher
Extensive concern.
Nano material small volume, cost is low, stable chemical performance, and good biocompatibility is safe, it is easy to prepare with protecting
Deposit, be most one of medicine carrying material of application value.Carbon quantum dot is that a kind of particle diameter is less than 10nm, carries the nanometer material of fluorescence
Material, with good water solubility, fluorescent stability, chemical inertness, biocompatibility, its surface carries various functions group and change
Key is learned, Covalent bonding together can be passed through with medicine or molecule.With the improvement to its synthesis technique, the preparing raw material of carbon quantum dot
Be more prone to obtain, it is nontoxic, preparation method is simpler, energy-saving and environmental protection.Carbon quantum dot is used as fluorescence probe, mark
Remember element and content of chemical substances inside and outside cell or detection organism.
New drug-loading system should possess load, release medicine；Fluorescence follows the trail of drug release process, research medicine effect machine
Reason；Several big features of the targeting for tumour cell.Many nano materials, such as CNT, graphene oxide, CdSe/ZnS quantum
Point, dendrimer etc., with preferable drug carrying ability, but due to material biological safety hidden danger, preparation technology is not simple enough, no
The problems such as possessing volume advantage, or do not possess fluorescence property, it is impossible to reach the best application demand of drug-loading system.Therefore new load
The research of drug material turns into the task of top priority.
The content of the invention
For above-mentioned deficiency of the prior art, adriamycin is loaded the invention provides a kind of biological safety carbon quantum dot
Complex and its preparation method and application, can effectively solve existing drug-loading system and there is potential safety hazard, preparation technology complexity etc. is asked
Topic.
To achieve the above object, the technical solution adopted for the present invention to solve the technical problems is：
A kind of biological safety carbon quantum dot loads adriamycin complex, and its preparation method comprises the following steps：
(1) it is 5-6 by volume by commercially available milk and deionized water:4-5 is mixed, and is subsequently placed in 160-200 DEG C of heating 2-
4h；
(2) step (1) resulting solution is naturally cooled into room temperature, then centrifuged under the conditions of 10000-12000r/min
30-45min, takes supernatant；
(3) supernatant obtained by step (2) is obtained into carbon quantum dot solution with 0.22 μm of membrane filtration；
(4) by carbon quantum dot solution vacuum freeze-drying obtained by step (3), deionized water is added, carbon quantum dot is made water-soluble
Liquid；
(5) doxorubicin hydrochloride powder is dissolved in deionized water, the adriamycin aqueous solution is made；
(6) the carbon quantum dot aqueous solution, step (5) the gained adriamycin aqueous solution and PBS solution obtained by step (4) are mixed,
PH=7.4 mixed solution is made；Wherein, doxorubicin concentration is 100-150 μ g/mL in mixed solution, and carbon quantum dot concentration is
1.5-4.5mg/mL；
(7) mixed solution obtained by step (6) is placed under the conditions of 100-200r/min, lucifuge reaction 12-24h, Ran Houtou
2-4h, the unsupported adriamycin of removing and carbon quantum dot are analysed, biological safety carbon quantum dot load adriamycin complex is obtained；Its
In, dialyzate is deionized water.
Further, it is 5 by volume by milk and deionized water in step (1):4 mixing, are subsequently placed in 180 DEG C of heating
2h。
Further, doxorubicin concentration is 100 μ g/mL in gained mixed solution in step (6), and carbon quantum dot concentration is
3mg/mL。
The biological safety carbon quantum dot load adriamycin complex of above-mentioned preparation can be substituted as new load medicine complex
Simple adriamycin, applied to tumor disease chemotherapeutic area.
A kind of biological safety carbon quantum dot load adriamycin complex that the present invention is provided and its preparation method and application,
Have the advantages that：
(1) carbon quantum dot carrier itself synthesis technique that the present invention is provided is simple, and advantages of nontoxic raw materials is harmless, cheap and easy to get；Medicine
Thing carrying method is simple, and condition requires relatively low, it is easy to operate, and repeatability is high, compared with other nano drug-carrying materials, is prepared into
This and implementation condition are lower.
(2) the biological safety carbon quantum dot load adriamycin complex that prepared by the present invention, with good fluorescent stabilization
Property, biological safety height, good biocompatibility；Carbon quantum dot can assist adriamycin preferably to be taken the photograph by cell by number of ways
Take.Compared to other non-fluorescence nano materials, the complex not only has good fluorescent stability, and volume and physicochemical property
Also it is preferable.
(3) the biological safety carbon quantum dot load adriamycin complex that prepared by the present invention, adriamycin load factor is high, reachable
87%, and then the dosage for carrying medicine complex is reduced, it is more efficiently convenient in actual applications；Meanwhile, adriamycin is released
Put with pH sensitiveness, the release rate highest under pH=5.0 environment, release rate is minimum in pH=7.4, can reach for swollen
Tumor tissue or cell administration, reduce the destruction of normal tissue, mitigate toxic side effect.
(4) the biological safety carbon quantum dot load adriamycin complex that prepared by the present invention, should compared to simple adriamycin
Complex is significantly improved to the killing-efficiency of tumour cell, the lethal obvious reduction to normal cell, complex can by Ah
Mycin passes into tumor cell core, destroys tumour cell nuclear dna, plays lethal effect, be favorably improved cancer
Therapeutic efficiency, reduces the adverse reaction of chemotherapeutics.
(5) complex has good fluorescence property, and the process of cellular uptake medicine can be traced, and positioning medicine enters cell
Position.
Brief description of the drawings
Fig. 1 is the transmission electron microscope (TEM) of carbon quantum dot (CDs) and carbon quantum dot load adriamycin complex (CDs-DOX),
ESEM (SEM), AFM (AFM) figure.
Fig. 2 is the fluorescence of carbon quantum dot (CDs) and biological safety carbon quantum dot load adriamycin complex (CDs-DOX)
Spectrogram and ultraviolet spectrogram.
Fig. 3 is that carbon quantum dot (CDs) and biological safety carbon quantum dot load the infrared of adriamycin complex (CDs-DOX)
Spectrogram, x-ray photon energy spectrum diagram and X-ray diffractogram.
Fig. 4 be simple adriamycin (DOX) and biological safety carbon quantum dot load adriamycin complex (CDs-DOX) with it is right
Salivary Adenoid Cystic Carcinoma Cell and the lethal testing result of l cell.
Fig. 5 is that biological safety carbon quantum dot loads copolymerization Jiao after adriamycin complex is absorbed by Salivary Adenoid Cystic Carcinoma Cell
Microscope imaging result.
Fig. 6 is that biological safety carbon quantum dot load adriamycin complex acts on tumour after Salivary Adenoid Cystic Carcinoma Cell, 48h
The result in cycle residing for cell.
Fig. 7 is under different pH environment, carbon quantum dot loads the release rate of adriamycin in adriamycin complex.
Embodiment
Embodiment 1
A kind of biological safety carbon quantum dot loads adriamycin complex, and its preparation method comprises the following steps：
(1) it is 5 by volume by commercially available milk and deionized water:4 mixing, are subsequently placed in 180 DEG C of heating 2h；
(2) step (1) resulting solution is naturally cooled into room temperature, 30min is then centrifuged under the conditions of 12000r/min, is taken
Supernatant；
(3) supernatant obtained by step (2) is obtained into carbon quantum dot solution with 0.22 μm of membrane filtration；
(4) by carbon quantum dot solution vacuum freeze-drying obtained by step (3), deionized water is added, the water-soluble of 12mg/mL is made
Liquid；
(5) doxorubicin hydrochloride powder is dissolved in deionized water, the 400 μ g/mL aqueous solution is made；
(6) the carbon quantum dot aqueous solution, step (5) the gained adriamycin aqueous solution and PBS solution obtained by step (4) are pressed into volume
Than for 1:1:2 mixing, are made pH=7.4 mixed solution；
(7) mixed solution obtained by step (6) is placed under the conditions of 200r/min, then lucifuge reaction 24h uses MWCO=
1000Da bag filter dialysis 2h (dialyzate is deionized water), the unsupported adriamycin of removing and carbon quantum dot, obtain biological peace
Full property carbon quantum dot load adriamycin complex.
Carbon quantum dot (CDs) and the biological safety carbon quantum dot of above-mentioned preparation load adriamycin complex (CDs-DOX)
Electron microscope as shown in figure 1, Fig. 1-1 and 1-2 be respectively CDs and CDs-DOX transmission electron microscope TEM figure, Fig. 1-3 and 1-4 difference
For CDs and CDs-DOX scanning electron microscope sem figure, Fig. 1-5 and 1-6 are respectively CDs and CDs-DOX atomic force microscope
Figure.As shown in Figure 1, the particle diameter of simple carbon quantum dot is in 5-10nm, highly in 1.5nm or so, and after having loaded adriamycin, carries
Medicine complex particle diameter, height and pattern are changed, and illustrate that adriamycin is successfully loaded to carbon quantum dot table by this method
Face.
The fluorescence spectrum of carbon quantum dot (CDs) and biological safety carbon quantum dot load adriamycin complex (CDs-DOX)
Figure and ultraviolet spectrogram are shown in Fig. 2；Wherein, Fig. 2-1 is fluorescence spectra, and Fig. 2-2 and 2-3 (Fig. 2-2 magnified partial view) are purple
External spectrum figure.As shown in Figure 2, the biological safety carbon quantum dot load adriamycin complex crest of synthesis there occurs movement, and
Show as blue shift i.e. crest and reduce direction movement to wave number, this phenomenon illustrates that biological safety carbon quantum dot load adriamycin is combined
Body is synthesized successfully.
The infrared spectrum of carbon quantum dot (CDs) and biological safety carbon quantum dot load adriamycin complex (CDs-DOX)
Figure, x-ray photon energy spectrum diagram and X-ray diffractogram are shown in Fig. 3；Wherein, Fig. 3-1 is infrared spectrogram, Fig. 3-2,3-3,3-4 and 3-
5 be x-ray photon energy spectrum diagram, and Fig. 3-6 is X-ray diffractogram.Infrared spectrogram shows, biological safety carbon quantum dot load Ah
Mycin complex possesses the characteristic peak of carbon quantum dot and adriamycin, but peak intensity and position there occurs certain change, together
When there is new characteristic peak to be formed compared to adriamycin.This shows that biological safety carbon quantum dot loads the successful conjunction of adriamycin complex
Into.X-ray photon energy spectrum diagram shows that biological safety carbon quantum dot load adriamycin complex is mainly made up of C, N, O, three kinds
The open score figure of element shows chemical bond present in load medicine complex, and the presence of C=C-N, C-N key can be biological with indirect proof
Security carbon quantum dot loads the successful synthesis of adriamycin complex.X-ray diffractogram shows, carbon quantum dot, adriamycin, biology
Security carbon quantum dot loads the roomy soft diffraction maximum of the equal only one of which of adriamycin complex, illustrates that three kinds of materials are amorphous
Body.
Embodiment 2
A kind of biological safety carbon quantum dot loads adriamycin complex, and its preparation method comprises the following steps：
(1) it is 1 by volume by commercially available milk and deionized water:1 mixing, is subsequently placed in 180 DEG C of heating 2h；
(2) step (1) resulting solution is naturally cooled into room temperature, 45min is then centrifuged under the conditions of 12000r/min, is taken
Supernatant；
(3) supernatant obtained by step (2) is obtained into carbon quantum dot solution with 0.22 μm of membrane filtration；
(4) by carbon quantum dot solution vacuum freeze-drying obtained by step (3), deionized water is added, the water-soluble of 12mg/mL is made
Liquid；
(5) doxorubicin hydrochloride powder is dissolved in deionized water, the 400 μ g/mL aqueous solution is made；
(6) the carbon quantum dot aqueous solution, step (5) the gained adriamycin aqueous solution and PBS solution obtained by step (4) are pressed into volume
Than for 1:2:5 mixing, are made pH=7.4 mixed solution；
(7) mixed solution obtained by step (6) is placed under the conditions of 200r/min, lucifuge reaction 24h is made.
Compared with Example 1, solution is partially red for product made from this method, and adriamycin residue is more.
Embodiment 3
A kind of biological safety carbon quantum dot loads adriamycin complex, and its preparation method comprises the following steps：
(1) it is 5 by volume by commercially available milk and deionized water:4 mixing, are subsequently placed in 180 DEG C of heating 2h；
(2) step (1) resulting solution is naturally cooled into room temperature, 30min is then centrifuged under the conditions of 12000r/min, is taken
Supernatant；
(3) supernatant obtained by step (2) is obtained into carbon quantum dot solution with 0.22 μm of membrane filtration；
(4) by carbon quantum dot solution vacuum freeze-drying obtained by step (3), deionized water is added, the water-soluble of 12mg/mL is made
Liquid；
(5) doxorubicin hydrochloride powder is dissolved in deionized water, the 400 μ g/mL aqueous solution is made；
(6) the carbon quantum dot aqueous solution, step (5) the gained adriamycin aqueous solution and PBS solution obtained by step (4) are pressed into volume
Than for 3:2:3 mixing, are made pH=7.4 mixed solution；
(7) mixed solution obtained by step (6) is placed under the conditions of 200r/min, lucifuge reaction 24h is made.
Compared with Example 1, solution is partially yellow for product made from this method, and adriamycin residue is more.
Embodiment 4
A kind of biological safety carbon quantum dot loads adriamycin complex, and its preparation method comprises the following steps：
(1) it is 5 by volume by commercially available milk and deionized water:4 mixing, are subsequently placed in 180 DEG C of heating 3h；
(2) step (1) resulting solution is naturally cooled into room temperature, 30min is then centrifuged under the conditions of 12000r/min, is taken
Supernatant；
(3) supernatant obtained by step (2) is obtained into carbon quantum dot solution with 0.22 μm of membrane filtration；
(4) by carbon quantum dot solution vacuum freeze-drying obtained by step (3), deionized water is added, the water-soluble of 12mg/mL is made
Liquid；
(5) doxorubicin hydrochloride powder is dissolved in deionized water, the 400 μ g/mL aqueous solution is made；
(6) the carbon quantum dot aqueous solution, step (5) the gained adriamycin aqueous solution and PBS solution obtained by step (4) are pressed into volume
Than for 1:1:2 mixing, are made pH=7.4 mixed solution；
(7) mixed solution obtained by step (6) is placed under the conditions of 200r/min, lucifuge reaction 12h is made.
Product made from this method is consistent with the outward appearance of embodiment 1.
Experimental example
Tested as follows using biological safety carbon quantum dot load adriamycin complex made from embodiment 1：
1st, lethal experiment of the product of the present invention to Salivary Adenoid Cystic Carcinoma Cell and l cell
Experimentation：
(1) Salivary Adenoid Cystic Carcinoma Cell, l cell are inoculated in 96 orifice plates respectively, and add 200 μ L and contain 10%
The RPMI culture mediums of hyclone, are containing 5%CO237 DEG C of constant temperature incubators in be incubated 24h, cell attachment；
(2) 5 μ L adriamycins, carbon quantum dot then are separately added into cell obtained by step (1) and load the compound of adriamycin
Body (so that the doxorubicin concentration in serum is 1.5 μM), is incubated 24h, 48h, 72h jointly, while setting the sky for being not added with medicine
White control group (being substituted with PBS)；
(3) it is incubated and terminates, remove culture medium, adds the RPMI containing 10%CCK-8 detection liquid but without hyclone and train
Base is supported, 1.5h is incubated；
(4) CCK-8 detections are carried out under 450nm excitation wavelengths, OD values is drawn, calculates cell survival rate, it is soft with SPSS
Part carries out statistical analysis to the difference group.
Killing experiments result is shown in that Fig. 4, wherein Fig. 4-1 are that adriamycin (DOX) loads adriamycin complex with carbon quantum dot
(CDs-DOX) to the lethal result figure of Salivary Adenoid Cystic Carcinoma Cell；Fig. 4-2 is that adriamycin (DOX) and carbon quantum dot load Ah are mould
Lethal result figure of the plain complex (CDs-DOX) to l cell.
As shown in Figure 4, the complex of carbon quantum dot load adriamycin the lethal effect of tumour cell is better than simple Ah
Mycin, is weaker than simple adriamycin to the lethal of normal cell.Analyze reason, thus it is speculated that can be helped after carbon quantum dot load adriamycin
Help adriamycin to enter neoplastic cell nuclei, improve the killing-efficiency of adriamycin；And adriamycin in itself to normal cell it is lethal just
To be less than after tumour cell, quantum dot load adriamycin, release rate of the adriamycin in normal cell is low, and release is slow, thus
Lethal further reduction.
2nd, nuclear targeting is tested
Experimentation：
(1) Salivary Adenoid Cystic Carcinoma Cell is inoculated in a diameter of 35nm culture dish, and adds 2mL containing 10% hyclone
RPMI culture mediums, containing 5%CO237 DEG C of constant temperature incubators in be incubated 24h, cell attachment；
(2) complex (CDs- that 100 μ L carbon quantum dots (CDs), adriamycin (DOX), carbon quantum dot load adriamycin is added
DOX) (so that carbon quantum dot concentration in serum is 300 μ g/mL, doxorubicin concentration is 3.5 μM) respectively with adenoid cystic carcinoma
Cell is incubated 4h jointly；
(3) remove culture medium and washed with PBS, 15min is fixed with 4% paraformaldehyde, PBS washings remove poly first
Aldehyde；
(4) experimental group that will be jointly incubated with carbon quantum dot (CDs) adds 6 μM of 1mL DiI cell membrane coloring agents, is shaking
15min is dyed in bed, PBS is washed 3 times；The μ g/mL of 1mL 1 DAPI nuclei dyeing toners are added, are dyed in shaking table
15min, PBS washing remove DAPI coloring agents, and the glycerine mounting for adding 10% is to be checked；
(5) experimental group that will be jointly incubated with simple adriamycin (DOX) adds 6 μM of 1mL DiO cell membrane coloring agents,
15min is dyed in shaking table, PBS is washed 3 times；The μ g/mL of 1mL 1 DAPI nuclei dyeing toners are added, are dyed in shaking table
15min, PBS washing remove DAPI coloring agents, and the glycerine mounting for adding 10% is to be checked；
(6) experimental group for being incubated the complex (CDs-DOX) that adriamycin is loaded with carbon quantum dot jointly adds 1mL
1 μ g/mL DAPI nuclei dyeing toners, dye 15min in shaking table, and PBS washings remove DAPI coloring agents, add 10% it is sweet
Sheet for oil seal is to be checked；
(7) cell dyeing situation is observed under Laser Scanning Confocal Microscope.
3 groups of cells are incubated after 4h jointly with Salivary Adenoid Cystic Carcinoma Cell, and its Laser Scanning Confocal Microscope imaging result figure is shown in Fig. 5.
As shown in Figure 5, simple carbon quantum dot can enter cell, be distributed mainly in cytoplasm；Simple adriamycin can
To enter cell, it is present in cytoplasm and nucleus, there is aggregation at nuclear membrane；Carbon quantum dot loads adriamycin group, into thin
The adriamycin showed increased of karyon, and focus primarily upon nucleus.This also demonstrates Fig. 4 experimental results, carbon quantum dot load
The complex of adriamycin, by acting on killing-efficiency of the nucleus raising to tumour cell.
3rd, experiment of the carbon quantum dot load adriamycin complex to Salivary Adenoid Cystic Carcinoma Cell cycle influences
Experimentation：
(1) l cell, Salivary Adenoid Cystic Carcinoma Cell are inoculated in a diameter of 35nm culture dish respectively, and added
Enter the RPMI culture mediums that 2mL contains 10% hyclone, contain 5%CO237 DEG C of constant temperature incubators in be incubated 24h, cell attachment；
(2) 100 μ L PBS, carbon quantum dot (CDs), simple adriamycin (DOX), carbon quantum dot are added and loads answering for adriamycin
Fit (CDs-DOX) (so that the carbon quantum dot concentration in serum is 200 μ g/mL, doxorubicin concentration is 1.5 μM) respectively with it is small
Mouse fibroblast cell, Salivary Adenoid Cystic Carcinoma Cell are incubated 48h jointly；
(3) it is incubated after terminating, centrifuges and be resuspended with PBS and washed, cell precipitation is obtained, with 100 μ L 1 × Annexin V
Binding solution is resuspended, and is subsequently added into 5 μ L Annexin V-FITC and 5 μ L propidium iodide (PI) coloring agent,
Lucifuge dyes 15min；
(4) cell cycle residing for each group cell is analyzed with flow cytomery.
PBS, carbon quantum dot (CDs), simple adriamycin (DOX), carbon quantum dot load the complex (CDs-DOX) of adriamycin
It is incubated jointly after 48h with l cell, Salivary Adenoid Cystic Carcinoma Cell respectively, cycle result residing for each group cell is shown in Fig. 6；Its
Middle Fig. 6-1,6-2,6-3 and 6-4 are respectively the knot of addition PBS, CDs, DOX and CDs-DOX in l cell (L929)
Fruit is schemed；6-5,6-6,6-7 and 6-8 are respectively addition PBS, CDs, DOX and CDs-DOX in Salivary Adenoid Cystic Carcinoma Cell (ACC-2)
Result figure.
It will be appreciated from fig. 6 that in l cell (L929), simple adriamycin group live cell fraction is about 80%, carbon amounts
Son point load adriamycin group live cell fraction rate is about 86%, slightly above simple adriamycin group.
In Salivary Adenoid Cystic Carcinoma Cell (ACC-2), simple adriamycin group living cells ratio is about 49%, carbon quantum dot load Ah
Mycin group live cell fraction rate is about 10%, and complex is significantly improved to the lethal of tumour cell；Simple adriamycin group is dead
Cell is about 46%, and the cells ratio in apoptosis late period is about 5%, and the two is than about 9：1, and carbon quantum dot load adriamycin
Group dead cell is about 68%, and the cells ratio in apoptosis late period is about 21%, and the two is than about 3：1, apoptosis ratio is obvious
Improve.As a result the mechanism for illustrating carbon quantum dot load adriamycin killing tumor cell may be the apoptosis for promoting tumour cell.
4th, the release experiment of adriamycin
External use PBS solution simulated in vivo environment, sets tri- groups of pH=5.0, pH=6.8 and pH=7.4 respectively, represents
Tumour cell endochylema, tumor tissues liquid and normal structure environment.The carbon quantum dot prepared load adriamycin complex is existed
Dialysed under three of the above pH environment, by measuring the fluorescence intensity of solution outside bag filter, and using doxorubicin fluorescence intensity with it is dense
Write music line, calculate the burst size (the adriamycin amount given) of adriamycin, you can obtain the release rate of adriamycin, i.e., it is unsupported
Adriamycin (A0), so as to extrapolate the load factor (DLE) of adriamycin.
Calculation formula is as follows：
DLE=(A1-A0)/A1
A1For the adriamycin total amount used in preparation process (6).
It is 87% by can be calculated adriamycin load factor.
Under different pH environment, the release rate result of adriamycin is shown in Fig. 7 in carbon quantum dot load adriamycin complex.By Fig. 7
Understand, adriamycin load factor is higher, and insoluble drug release has pH sensitiveness, the release rate highest under pH5.0 environment, is released in pH7.4
Put rate minimum, the side effect for reducing normal tissue for tumor tissues or cell administration can be reached；Complex has certain
Slow releasing function, discharge after 10h and gradually tend to be steady, monitoring is still slowly discharging to 72h.
Claims (5)
1. a kind of biological safety carbon quantum dot loads the preparation method of adriamycin complex, it is characterised in that including following step
Suddenly：
(1) it is 5-6 by volume by milk and deionized water:4-5 is mixed, and is subsequently placed in 160-200 DEG C of heating 2-4h；
(2) step (1) resulting solution is naturally cooled into room temperature, 30- is then centrifuged under the conditions of 10000-12000r/min
45min, takes supernatant；
(3) supernatant obtained by step (2) is obtained into carbon quantum dot solution with 0.22 μm of membrane filtration；
(4) by carbon quantum dot solution vacuum freeze-drying obtained by step (3), deionized water is added, the carbon quantum dot aqueous solution is made；
(5) doxorubicin hydrochloride powder is dissolved in deionized water, the adriamycin aqueous solution is made；
(6) the carbon quantum dot aqueous solution, step (5) the gained adriamycin aqueous solution and PBS solution obtained by step (4) are mixed, are made
PH=7.4 mixed solution；Wherein, doxorubicin concentration is 100-150 μ g/mL in mixed solution, and carbon quantum dot concentration is 1.5-
4.5mg/mL；
(7) mixed solution obtained by step (6) is placed under the conditions of 100-200r/min, lucifuge reaction 12-24h, then dialyse 2-
4h, the unsupported adriamycin of removing and carbon quantum dot, obtain biological safety carbon quantum dot load adriamycin complex；Wherein, thoroughly
Analysis liquid is deionized water.
2. biological safety carbon quantum dot according to claim 1 loads the preparation method of adriamycin complex, its feature
Be, in step (1) by milk and deionized water by volume be 5:4 mixing, are subsequently placed in 180 DEG C of heating 2h.
3. biological safety carbon quantum dot according to claim 1 loads the preparation method of adriamycin complex, its feature
It is, doxorubicin concentration is 100 μ g/mL in gained mixed solution in step (6), carbon quantum dot concentration is 3mg/mL.
4. the biological safety carbon quantum dot as described in claim any one of 1-3 loads the preparation method system of adriamycin complex
For the biological safety carbon quantum dot load adriamycin complex gone out.
5. biological safety carbon quantum dot load adriamycin complex as claimed in claim 4 is in terms of tumor disease is treated
Using.
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