CN110237050A - A kind of Combretastatin nanoparticle and preparation method thereof - Google Patents
A kind of Combretastatin nanoparticle and preparation method thereof Download PDFInfo
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/075—Ethers or acetals
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- A61K31/09—Ethers or acetals having an ether linkage to aromatic ring nuclear carbon having two or more such linkages
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- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5107—Excipients; Inactive ingredients
- A61K9/513—Organic macromolecular compounds; Dendrimers
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Abstract
The present invention relates to a kind of Combretastatin (CA4) nanoparticles and preparation method thereof, comprising the following steps: surfactant-dispersed Yu Shuizhong is stirred and be preheated to 35-45 DEG C, as water phase by (a)；(b) CA4 and degradable polymer are dissolved in organic solvent, as oily phase；And (c) under stiring, oil is added in water phase, volatile organic solvent under the conditions of 35-45 DEG C of emulsifying temperature obtains CA4 nanoparticle.The present invention prepares CA4 nanoparticle using O/W type emulsification-evaporation method, and preferably, encapsulation rate is high for dispersibility, and uniform particle sizes are suitable for, and have well solved the low problem of CA4 solubility, obtain the preparation with slow release effect.
Description
Technical field
The present invention relates to field of pharmaceutical preparations more particularly to a kind of Combretastatin nanoparticle and preparation method thereof.
Background technique
Cancer originates from epithelial tissue, is one kind under the action of physics, chemistry and/or inherent cause, causes former cancer base
Cause and/or tumor suppressor gene mutate, and cause normal cell metabolism to regulate and control disorder, natural death of cerebral cells imbalance, cell mitogen is not
Modulated malignant tumour.The unlimited growth of cancer cell can destroy normal surrounding tissue, so that organ, bone, blood etc. lose
Normal function damages human health, final threat to life.
Currently, the treatment method of malignant tumour mainly has chemotherapy, radiotherapy, operation excision.But at present
For clinical anticancer drug have mostly normal tissue toxicity it is big, it is ineffective to tumorcidal, easily supported by tumour cell
The disadvantages of anti-.So the research of cancer treatment drugs is a current hot spot.
Combretastatin (CA4) is a kind of novel vasoinhibitor, is one kind cis- two similar to the structure of colchicin
Phenylethylene native compound, molecular weight 316.35, CA4 is using the tubulin of tumor vascular endothelial cell as target spot, with the autumn
Tazettine has identical tubulin binding site, and binding ability is stronger, is able to suppress tubulin polymerization, and cell is prevented to have
Silk division finally makes tumour cell dead due to nutritional deficiency to inhibit vascularization.It has the inhibiting tumor cell of wide spectrum living
Property, on kinds of tumors model all effectively.Also, the dosage that CA4 generates antitumor action is that it generates toxicity dose very
One of, it is small on normal cell influence, it is a kind of novel anti-tumor drug.
But since there is tubulins in the cell of human body, which results in CA4 to the inhibiting effect of tumour cell
Specificity is not strong, and injury can be also generated to human normal cell, when large dosage in use, can also endanger cardiovascular and maincenter mind
It is lipophilic compound through system also, CA4, is difficult to dissolve in aqueous solution, leading to it, curative effect is lower in vivo, not to remnants
The tumor tissues lethality of kill is weaker, make it clinically application be greatly limited.Therefore, CA4 is improved in body
Interior curative effect and the injury for reducing its normal tissue, are the emphasis studied at present.
For this purpose, researchers have carried out various modifications to CA4 and prodrug is studied, wherein Combretastatin disodium hydrogen phosphate (CA4P)
It is great representative one kind, the external medicine has entered II/III phase clinical investigation phase.CA4P passes through intravenous injection administration, energy
Effect in human body through enzyme is converted to CA4 and plays a role.But CA4P there are toxic side effects big, internal residence time
Short, the disadvantages of stability is poor, and its light and it is heated after easily decompose.Nitrogen-containing group is introduced into CA4 in addition, going back some researchers
To improve its solubility.In the compound of synthesis, replace the compound of the phenolic hydroxyl group of CA4 small to CT-26 in vitro with amino
Mouse adenocarcinoma of colon tumour cell has stronger anti-angiogenic protein active and cytotoxicity.Some other synthesis effective in vitro
Object, experimenter have carried out internal assessment to them, and it is living that some of them synthetic shows apparent anticancer in animal model
Property, the anticancer activity and CDDP of these compounds are quite or better than CDDP, but the internal safety of these compounds does not determine yet.
Then, researchers both domestic and external accelerate to nanotechnology course of drug development research, it is desirable to by change dosage form come
Improve clinical drug using effect.
In terms of containing CA4 targeting preparation research, CA4 is encapsulated in by Zhang Yalan etc. using Emulsion evaporation-solidification at low temperature
In solid lipid nano preparation, there is apparent slow releasing function in vitro, but its curative effect in vivo need further to investigate.
Yi-fei Zhang etc. successfully prepares and characterizes the PEG-PLA microballoon for containing CA4 modified with RGD.RGD targeting
The endocytosis that microballoon is mediated by cascade albumen enhances absorption of the tumor vascular endothelial cell to entrapped drug, makes CA4 more
It plays a role well.
Zhang etc. has synthesized the G5 polyamide dendrimer of the fluorescein isothiocynate and modified with folic acid that contain CA4
(G5.NHAc-FI-FA/CA4), the solubility of CA4 in aqueous solution is increased to 240 μ g/mL from 11.8 μ g/mL significantly, and
And stability is enhanced, enable CA4 with constant, slow rate release.This kind of inclusion compound can be specifically targeted folacin receptor
The tumour cell of overexpression, but also need further in vivo studies.
The poloxamer F127 preparation of the ethylenediamine crosslinking of 76% polyethylene oxide block such as Rajesh R.Wakaskar
A kind of polymer micelle containing CA4.This micella reduces the premature disconnection excessively of drug in blood compared with the aqueous solution of CA4
It puts, increases mean residence time of the drug in blood plasma.
There are also researchers to be integrated to CA4 in three block tree shaped polymer, and this tree shaped polymer contains hydrophilic PEG
Side chain and the hydrophobic polyester tree shaped polymer that can contain CA4, can biodegrade.The variation of tree shaped polymer parting can
To be used to obtain the micrograined texture of different size and different drugloading rates.The hydrophily of this existing PEG of structure, it is dirty without biology
Dye, is also equipped with the biodegradability of tree shaped polymer.This particle, which possesses, is suitble to the fluid dynamics of passive cancer target straight
Diameter is good pharmaceutical controlled release formulation.After it is loaded with CA4, the survival rate of normal cell is significantly improved, and show compared with
Good curative effect.
In conclusion the application of nanotechnology can reduce poisonous side effect of medicine, Drug controlled release is improved in drug body and is treated
Effect, and there is test to prove CA4 nanometer formulation to the inhibiting rate of tumour better than CA4P.But due to the metabolite of nano material
There are certain cytotoxicity, lead to not confirm its safety.
Summary of the invention
In order to solve the above technical problems, the object of the present invention is to provide a kind of CA4 nanoparticle and preparation method thereof, the present invention
CA4 nanoparticle is prepared using O/W emulsification-evaporation method, play the role of solubilized CA4 and is sustained.To achieve the goals above,
The present invention provides the following technical solutions:
The first purpose of the invention is to provide a kind of preparation methods of Combretastatin nano-particle composition, including following step
It is rapid:
(a) it by surfactant-dispersed Yu Shuizhong, stirs and is preheated to 35-45 DEG C, as water phase；
(b) CA4 and degradable polymer are dissolved in organic solvent, as oily phase；
And (c) under stiring, oil is added in water phase, volatile organic solvent under the conditions of 35-45 DEG C of emulsifying temperature obtains
To CA4 nanoparticle.
Further, in step (a), the surfactant is selected from phosphide class, polyol type copolymer, polyoxyethylene
One or more of type copolymer and Pluronic F68.
Preferably, surfactant is selected from phospholipid.It is highly preferred that surfactant is soybean lecithin S100.
Further, in step (a), the surfactant accounts for the 0.001% (w/v) -1% (w/v) of water phase.
Preferably, surfactant accounts for the 0.003% (w/v) -0.05% (w/v) of water phase.
Further, in step (b), the degradable polymer be selected from lactide-glycolide copolymer, polylactic acid,
One or more of polycaprolactone and polylactic acid-polyglycol.
Preferably, degradable polymer is selected from poly lactide-glycolide acid or polylactic acid.Poly lactic-co-glycolic acid
Monomer ratio in copolymer is 50:50~85:15, the molecular weight of poly lactide-glycolide acid is 10000~
60000g/mol。
Further, in step (b), the mass ratio of the CA4 and degradable polymer is 1:1-1:100.
Preferably, the mass ratio of CA4 and degradable polymer is 1:1-1:10.
Further, in step (b), Combretastatin accounts for the 0.5-5% (w/v) of oily phase.
Further, in step (b), it is sub- that the organic solvent is selected from acetone, methylene chloride, ethyl acetate, dimethyl
One or more of sulfone and methyl ethyl ketone.
Preferably, organic solvent is acetone or dimethyl sulfoxide.It is highly preferred that organic solvent is acetone.
Further, in step (c), the oil is mutually 1:2-1:100 with the volume ratio of water phase.
Preferably, oil is mutually 1:5-1:50 with the ratio of water phase.
Further, in step (c), mixing speed 800-1600rpm.
A second object of the present invention is to provide a kind of using CA4 nanoparticle prepared by above-mentioned preparation method, including CA4
And it is wrapped in the surface CA4 degradable polymer, the meso-position radius of the nanoparticle is 50~300nm, the degradable polymer
Energy solubilising CA4, and there is the feature of sustained release.
According to the above aspect of the present invention, the present invention has at least the following advantages:
The present invention prepares CA4 nanoparticle using O/W type emulsification-evaporation method.Firstly, using polymer for degradable poly
Close object, good biocompatibility；Secondly, using the ratio of reasonable surfactant, suitable material ratio and water phase and oily phase,
Obtain preferable nanoparticle, good dispersion, encapsulation rate is high, and uniform particle sizes are suitable for, and has well solved that CA4 solubility is low to ask
Topic, obtains the preparation with slow release effect.
The above description is only an overview of the technical scheme of the present invention, in order to better understand the technical means of the present invention,
And can be implemented in accordance with the contents of the specification, the following is a detailed description of the preferred embodiments of the present invention and the accompanying drawings.
Detailed description of the invention
Fig. 1 is the dispersion figure of the nanoparticle of embodiment 4-12 preparation；
Fig. 2 is the transmission electron microscope picture of CA4 nanoparticle prepared by embodiment 4；
Fig. 3 is the transmission electron microscope picture of CA4 nanoparticle prepared by embodiment 5；
The In-vitro release curves for the CA4 nanoparticle that Fig. 4 is CA4 and prepared by embodiment 13.
Specific embodiment
With reference to the accompanying drawings and examples, specific embodiments of the present invention will be described in further detail.Implement below
Example is not intended to limit the scope of the invention for illustrating the present invention.
Embodiment 1
Precision weighs soybean lecithin 7.5mg and is dissolved in 1mL dehydrated alcohol, is configured to the soybean lecithin of 7.5mg/mL
S100 mother liquor.25 μ L soybean lecithin S100 mother liquors are taken, are added in the deionized water of 5mL, are placed in magnetic stirring apparatus, and preheat
To 37 DEG C, as water phase.Precision weighs PLGA 12.5mg and is dissolved in 1mL acetone, is configured to the PLGA mother liquor of 12.5mg/mL.
100 μ LPLGA mother liquors are taken, 250 μ L is diluted to acetone, is added in the water phase under stirring, mixing speed is
1600rpm, is then slowly stirred 3h under the conditions of 40 DEG C, and volatile organic solvent obtains blank nanoparticle suspension.
Embodiment 2
Precision weighs soybean lecithin 7.5mg and is dissolved in 1mL dehydrated alcohol, is configured to the soybean lecithin of 7.5mg/mL
S100 mother liquor.100 μ L soybean lecithin S100 mother liquors are taken, are added in the deionized water of 5mL, are placed in magnetic stirring apparatus, and is pre-
Heat is to 37 DEG C, as water phase.Precision weighs PLGA 12.5mg and is dissolved in 1mL acetone, and the PLGA for being configured to 12.5mg/mL is female
Liquid.100 μ LPLGA mother liquors are taken, 250 μ L is diluted to acetone, is added in the water phase under stirring, mixing speed is
1600rpm, is then slowly stirred 3h under the conditions of 40 DEG C, and volatile organic solvent obtains blank nanoparticle suspension.
Embodiment 3
Precision weighs soybean lecithin 500mg and is dissolved in 6mL dehydrated alcohol, is added in the deionized water of 100mL, places
In magnetic stirring apparatus, and 37 DEG C are preheating to, as water phase.Precision weighs PLA 12.5mg and is dissolved in 1mL dimethyl sulfoxide, configuration
At the PLA mother liquor of 12.5mg/mL.Precision weighs CA4 10.0mg and is dissolved in 1mL acetone, and the CA4 for being configured to 10.0mg/mL is female
Liquid.1mL PLA mother liquor is taken, 2.5mL is diluted to acetone, 12.5 μ L CA4 mother liquors is added, mix, be added under stirring
Water phase in, then mixing speed 800rpm is slowly stirred 3h under the conditions of 45 DEG C, volatile organic solvent obtains CA4 nanometers
Grain suspension.
Embodiment 4
Precision weighs soybean lecithin 7.5mg and is dissolved in 1mL dehydrated alcohol, is configured to the soybean lecithin of 7.5mg/mL
S100 mother liquor.50 μ L soybean lecithin S100 mother liquors are taken, are added in the deionized water of 5mL, are placed in magnetic stirring apparatus, and preheat
To 37 DEG C, as water phase.Precision weighs PLGA 12.5mg and is dissolved in 1mL acetone, is configured to the PLGA mother liquor of 12.5mg/mL.
100 μ LPLGA mother liquors are taken, 250 μ L is diluted to acetone, is added in the water phase under stirring, mixing speed is
1200rpm, is then slowly stirred 3h under the conditions of 40 DEG C, and volatile organic solvent obtains blank nanoparticle suspension.
Embodiment 5
Precision weighs soybean lecithin 7.5mg and is dissolved in 1mL dehydrated alcohol, is configured to the soybean lecithin of 7.5mg/mL
S100 mother liquor.50 μ L soybean lecithin S100 mother liquors are taken, are added in the deionized water of 5mL, are placed in magnetic stirring apparatus, and preheat
To 37 DEG C, as water phase.Precision weighs PLGA 12.5mg and is dissolved in 1mL acetone, is configured to the PLGA mother liquor of 12.5mg/mL.
Precision weighs CA4 10.0mg and is dissolved in 1mL acetone, is configured to the CA4 mother liquor of 10.0mg/mL.100 μ L PLGA mother liquors are taken,
250 μ L are diluted to acetone, 25 μ L CA4 mother liquors are added, mixes, is added in the water phase under stirring, mixing speed is
1200rpm, is then slowly stirred 3h under the conditions of 37 DEG C, and volatile organic solvent obtains CA4 nanoparticle suspension.
Embodiment 6
Precision weighs soybean lecithin 7.5mg and is dissolved in 1mL dehydrated alcohol, is configured to the soybean lecithin of 7.5mg/mL
S100 mother liquor.50 μ L soybean lecithin S100 mother liquors are taken, are added in the deionized water of 5mL, are placed in magnetic stirring apparatus, and preheat
To 37 DEG C, as water phase.Precision weighs PLGA 12.5mg and is dissolved in 1mL acetone, is configured to the PLGA mother liquor of 12.5mg/mL.
Precision weighs CA4 10.0mg and is dissolved in 1mL acetone, is configured to the CA4 mother liquor of 10.0mg/mL.100 μ L PLGA mother liquors are taken,
250 μ L are diluted to acetone, 50 μ L CA4 mother liquors are added, mixes, is added in the water phase under stirring, mixing speed is
1200rpm, is then slowly stirred 3h under the conditions of 37 DEG C, and volatile organic solvent obtains CA4 nanoparticle suspension.
Embodiment 7
Precision weighs soybean lecithin 7.5mg and is dissolved in 1mL dehydrated alcohol, is configured to the soybean lecithin of 7.5mg/mL
S100 mother liquor.50 μ L soybean lecithin S100 mother liquors are taken, are added in the deionized water of 5mL, are placed in magnetic stirring apparatus, and preheat
To 37 DEG C, as water phase.Precision weighs PLGA 12.5mg and is dissolved in 1mL acetone, is configured to the PLGA mother liquor of 12.5mg/mL.
200 μ L PLGA mother liquors are taken, 500 μ L is diluted to acetone, is added in the water phase under stirring, mixing speed is
1200rpm, is then slowly stirred 3h under the conditions of 37 DEG C, and volatile organic solvent obtains blank nanoparticle suspension.
Embodiment 8
Precision weighs soybean lecithin 7.5mg and is dissolved in 1mL dehydrated alcohol, is configured to the soybean lecithin of 7.5mg/mL
S100 mother liquor.50 μ L soybean lecithin S100 mother liquors are taken, are added in the deionized water of 5mL, are placed in magnetic stirring apparatus, and preheat
To 37 DEG C, as water phase.Precision weighs PLGA 12.5mg and is dissolved in 1mL acetone, is configured to the PLGA mother liquor of 12.5mg/mL.
Precision weighs CA4 10.0mg and is dissolved in 1mL acetone, is configured to the CA4 mother liquor of 10.0mg/mL.200 μ L PLGA mother liquors are taken,
500 μ L are diluted to acetone, 50 μ L CA4 mother liquors are added, mixes, is added in the water phase under stirring, mixing speed is
1200rpm, is then slowly stirred 3h under the conditions of 37 DEG C, and volatile organic solvent obtains CA4 nanoparticle suspension.
Embodiment 9
Precision weighs soybean lecithin 7.5mg and is dissolved in 1mL dehydrated alcohol, is configured to the soybean lecithin of 7.5mg/mL
S100 mother liquor.50 μ L soybean lecithin S100 mother liquors are taken, are added in the deionized water of 5mL, are placed in magnetic stirring apparatus, and preheat
To 37 DEG C, as water phase.Precision weighs PLGA 12.5mg and is dissolved in 1mL acetone, is configured to the PLGA mother liquor of 12.5mg/mL.
Precision weighs CA4 10.0mg and is dissolved in 1mL acetone, is configured to the CA4 mother liquor of 10.0mg/mL.200 μ L PLGA mother liquors are taken,
500 μ L are diluted to acetone, 100 μ L CA4 mother liquors are added, mixes, is added in the water phase under stirring, mixing speed
For 1200rpm, 3h is then slowly stirred under the conditions of 37 DEG C, volatile organic solvent obtains CA4 nanoparticle suspension.
Embodiment 10
Precision weighs soybean lecithin 7.5mg and is dissolved in 1mL dehydrated alcohol, is configured to the soybean lecithin of 7.5mg/mL
S100 mother liquor.50 μ L soybean lecithin S100 mother liquors are taken, are added in the deionized water of 5mL, are placed in magnetic stirring apparatus, and preheat
To 37 DEG C, as water phase.Precision weighs PLGA 12.5mg and is dissolved in 1mL acetone, is configured to the PLGA mother liquor of 12.5mg/mL.
400 μ L PLGA mother liquors are taken, 1000 μ L is diluted to acetone, is added in the water phase under stirring, mixing speed is
1200rpm, is then slowly stirred 3h under the conditions of 37 DEG C, and volatile organic solvent obtains blank nanoparticle suspension.
Embodiment 11
Precision weighs soybean lecithin 7.5mg and is dissolved in 1mL dehydrated alcohol, is configured to the soybean lecithin of 7.5mg/mL
S100 mother liquor.50 μ L soybean lecithin S100 mother liquors are taken, are added in the deionized water of 5mL, are placed in magnetic stirring apparatus, and preheat
To 37 DEG C, as water phase.Precision weighs PLGA 12.5mg and is dissolved in 1mL acetone, is configured to the PLGA mother liquor of 12.5mg/mL.
Precision weighs CA4 10.0mg and is dissolved in 1mL acetone, is configured to the CA4 mother liquor of 10.0mg/mL.400 μ L PLGA mother liquors are taken,
1000 μ L are diluted to acetone, 100 μ L CA4 mother liquors are added, mixes, is added in the water phase under stirring, mixing speed
For 1200rpm, 3h is then slowly stirred under the conditions of 37 DEG C, volatile organic solvent obtains CA4 nanoparticle suspension.
Embodiment 12
Precision weighs soybean lecithin 7.5mg and is dissolved in 1mL dehydrated alcohol, is configured to the soybean lecithin of 7.5mg/mL
S100 mother liquor.50 μ L soybean lecithin S100 mother liquors are taken, are added in the deionized water of 5mL, are placed in magnetic stirring apparatus, and preheat
To 37 DEG C, as water phase.Precision weighs PLGA 12.5mg and is dissolved in 1mL acetone, is configured to the PLGA mother liquor of 12.5mg/mL.
Precision weighs CA4 10.0mg and is dissolved in 1mL acetone, is configured to the CA4 mother liquor of 10.0mg/mL.400 μ L PLGA mother liquors are taken,
1000 μ L are diluted to acetone, 200 μ L CA4 mother liquors are added, mixes, is added in the water phase under stirring, mixing speed
For 1200rpm, 3h is then slowly stirred under the conditions of 37 DEG C, volatile organic solvent obtains CA4 nanoparticle suspension.
Embodiment 13
Precision weighs soybean lecithin 7.5mg and is dissolved in 1mL dehydrated alcohol, is configured to the soybean lecithin of 7.5mg/mL
S100 mother liquor.200 μ L soybean lecithin S100 mother liquors are taken, are added in the deionized water of 20mL, are placed in magnetic stirring apparatus, and is pre-
Heat is to 37 DEG C, as water phase.Precision weighs PLGA 12.5mg and is dissolved in 1mL acetone, and the PLGA for being configured to 12.5mg/mL is female
Liquid.Precision weighs CA4 10.0mg and is dissolved in 1mL acetone, is configured to the CA4 mother liquor of 10.0mg/mL.Take 400 μ L PLGA female
Liquid is diluted to 1000 μ L with acetone, 100 μ L CA4 mother liquors is added, mix, be added in the water phase under stirring, stirs
Speed is 1200rpm, and 3h is then slowly stirred under the conditions of 37 DEG C, and volatile organic solvent obtains CA4 nanoparticle suspension.With
Afterwards, the CA4 nanoparticle suspension in beaker is collected, 30min is centrifuged under low temperature (4 DEG C) high-speed condition, discards supernatant liquid, use is ultrapure
Water cleans the CA4 nanoparticle precipitating of bottom, in triplicate, finally with ultrapure water dispersion to get CA4 nanoparticle.It is to freeze with glycerol
Dry protective agent, freeze-drying, freeze temperature are -50 DEG C, and freeze-drying pressure is 0.52mbar to get CA4 nano-granule freeze-dried powder.
The test of 1 nanoparticle partial size of test case
The nanoparticle suspension for drawing the preparation of 200 μ L embodiments 1,2,4,5,6 respectively, is diluted to 2mL with ultrapure water, is placed in
The partial size of sample is measured in laser particle size analyzer.It the results are shown in Table 1, resulting microspherulite diameter (refering in particular to meso-position radius) range is in 100-
Between 200nm.
The microspherulite diameter of the different embodiment preparations of table 1
| Embodiment | Mass median diameter |
| 1 | 185.6 |
| 2 | 167.2 |
| 3 | 203.4 |
| 4 | 117.3 |
| 5 | 162.5 |
| 6 | 145.8 |
| 7 | 120.6 |
| 8 | 173.2 |
| 9 | 169.8 |
| 10 | 134.7 |
| 11 | 178.4 |
| 12 | 167.1 |
| 13 | 154.7 |
2 nanoparticle dispersion of test case
After embodiment 4-12 is prepared resulting nanoparticle suspension ultrasonic disperse, a period of time is stood, observation dispersion feelings
Condition.The result is shown in Figure 1 in Fig. 1 a-c, is corresponding in turn to nanoparticle prepared by embodiment 4-12 from left to right, the results showed that, this hair
The dispersibility of bright CA4 nanoparticle and blank nanoparticle is close, and dispersibility is preferably.
The test of 3 nanoparticle form of test case
Example 4, embodiment 5 prepare resulting nanoparticle suspension, and ultrasonic disperse is at liquid condition.200 mesh are micro-
Grid are lain in a horizontal plane on filter paper, and sample is added dropwise in micro-grid rapidly, are placed under infrared lamp, and dry 30min reuses hair dryer
Blow away the nanoparticle sample not being attached on copper sheet, using transmission electron microscope observation nanoparticle shape characteristic and record figure
Piece.As a result see Fig. 2, Fig. 3.The result shows that the uniform particle sizes of CA4 nanoparticle of the invention and blank nanoparticle are suitable for, form rule
It is whole.
4 nanoparticle drugloading rate of test case and encapsulation rate are tested
Embodiment 5, embodiment 6 are prepared into resulting CA4 nanoparticle suspension, are fitted into ultra-filtration centrifuge tube, in 8000g, 4
Under the conditions of DEG C, ultracentrifugation 30min collects supernatant, measures the content of free drug in outer aqueous phase.It is calculated according to the following formula and receives
The encapsulation rate (Entrapment efficiency, EE) and drugloading rate (Loading efficiency, LE) of the grain of rice, are as a result shown in
Table 2 (n=3).
EE=(WAlways-WIt is free)/WAlways× 100%
LE=(WAlways-WIt is free)/WCarrier× 100%
(wherein, WAlwaysIt is drug total amount, WIt is freeIt is the amount that do not wrap into the drug of nanoparticle, WCarrierIt is the quality of carrier in nanometer)
The drugloading rate and encapsulation rate of table 2CA4 nanoparticle
| Embodiment | Drugloading rate (%) ± SD | Encapsulation rate (%) ± SD |
| 5 | 17.88±0.02 | 89.38±0.08 |
| 6 | 28.44±0.08 | 71.09±0.18 |
The In-vitro release curves of 5 CA4 nanoparticle of test case are tested
The phosphate buffer solution of pH 7.4 (0.5%Tween 80) is chosen as dissolution medium, investigates drug-carrying nanometer particle
Release characteristics.Each 3 parts of CA4, CA4 nanoparticle (embodiment 13, CA4NPs) for taking phase homogenous quantities, respectively by CA4, embodiment 13
The CA4 nanoparticle of preparation is concentrated into 5mL (CA4=1.5mg), is subsequently placed in processed bag filter (MWCO=3000),
It tightens, is suspended in the conical flask for filling the phosphate buffer of 25mL, every group of parallel three sample, at 37 DEG C, 200 × g item
Extracorporeal releasing test is carried out under part.And in 4h, 8h, 12h, for 24 hours, 36h, 48h and 72h when, sample 2mL, while adding isometric
Synthermal dissolution medium.Samples taken 12000 × g high speed centrifugation 10min, take supernatant with high phase liquid chromatography for measuring its
Burst size, and its cumulative release amount is calculated, draw CA4 In-vitro release curves.As a result see Fig. 4.
It can be seen from the figure that CA4 is only capable of release 41.5% because of itself poor solubility, in 72h, and when it is prepared into
After CA4 nanoparticle (CA4NPs), it can be discharged into 81.5%, illustrate that nanoparticle, which is made, can increase the solubility of CA4, and can reach
Slow releasing function.
The above is only a preferred embodiment of the present invention, it is not intended to restrict the invention, it is noted that for this skill
For the those of ordinary skill in art field, without departing from the technical principles of the invention, can also make it is several improvement and
Modification, these improvements and modifications also should be regarded as protection scope of the present invention.
Claims (9)
1. a kind of preparation method of Combretastatin nanoparticle, which comprises the following steps:
(a) it by surfactant-dispersed Yu Shuizhong, stirs and is preheated to 35-45 DEG C, as water phase；
(b) Combretastatin and degradable polymer are dissolved in organic solvent, as oily phase；
And (c) under agitation, oil is added in water phase, volatile organic solvent under the conditions of 35-45 DEG C of emulsifying temperature obtains
To Combretastatin nanoparticle.
2. preparation method according to claim 1, it is characterised in that: in step (a), the surfactant is selected from phosphorus
One or more of lipid, polyol type copolymer, polyoxyethylene-type copolymer and Pluronic F68.
3. preparation method according to claim 1, it is characterised in that: in step (b), the degradable polymer is selected from
One or more of poly lactide-glycolide acid, polylactic acid, polycaprolactone and polylactic acid-polyglycol.
4. preparation method according to claim 1, it is characterised in that: in step (b), the organic solvent is selected from third
One or more of ketone, methylene chloride, ethyl acetate, dimethyl sulfoxide and methyl ethyl ketone.
5. preparation method according to claim 1, it is characterised in that: in step (a), the surfactant accounts for water phase
0.001% (w/v) -1% (w/v).
6. preparation method according to claim 1, it is characterised in that: in step (b), the Combretastatin with it is degradable
The mass ratio of polymer is 1:1-1:100.
7. preparation method according to claim 1, it is characterised in that: in step (b), the Combretastatin accounts for oily phase
0.5-5% (w/v).
8. preparation method according to claim 1, it is characterised in that: in step (c), the volume of the oil phase and water phase
Than for 1:2-1:100.
9. Combretastatin nanoparticle prepared by a kind of preparation method of any of claims 1-8, it is characterised in that:
Including Combretastatin and it is wrapped in the degradable polymer on Combretastatin surface, the meso-position radius of the Combretastatin nanoparticle is
50-300nm。
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