A kind of preparation method for the PLGA biodegradable microspheres loading disulfiram drug
Technical field
The present invention relates to a kind of preparation method of PLGA biodegradable microspheres, in particular to a kind of load disulfiram drug
The preparation method of PLGA biodegradable microspheres, belongs to field of biomedical polymer materials.
Background technique
Disulfiram (disulfiram, DSF) is used for more than 60 years as treatment alcohol addiction disease drug, is studied recently
It was found that disulfiram has very high specificity to tumour cell, tumour cell is only killed, normal cell is not injured.DSF energy
Tumor stem cell is killed in targeting, also not fully aware of at present to the mechanism of action of tumor stem cell.Disulfide bond in DSF molecule
It is easy to be destroyed by gluathione peptides reducing agent during blood transportation, half-life period in blood less than 4 minutes, makes DSF
It has just been inactivated before tumor tissues have not yet been reached.Nanoparticle-loaded DSF can slow down its degradation inactivation.Pass through liposome
Package can extend the half-life period of DSF in vivo, however 18 minutes half-life period of DSF liposome from clinical application there are also it is very long away from
From.Distinctive parent-hydrophobic the core-shell structure of macromolecule micelle can be effectively protected during blood conveys be carried on it is hydrophobic
DSF in core.The DSF of poly lactide-glycolide acid (PLGA) nanoparticle package has the half-life period up to 4 hours.
Due to the limited space in inside of DSF strong hydrophobicity and nanoparticle, it is difficult to largely load DSF.Document report
Drugloading rate of the DSF in nanoparticle it is all very low, generally 3% or less.PLGA microballoon has hollow spherical shell structure, wherein
Empty internal structure is greatly improved the load capacity of DSF drug.The PLGA microsphere surface of conventional emulsification method preparation is adsorbed with largely
Surfactant, although the surfactants of partial adsorbates can be removed by repeatedly washing, remove and be not thorough.
Surfactant can adsorb the DSF not being wrapping to inside microballoon, cause the drugloading rate of DSF higher, error is up to 50% or more.
Surfactant can not be had to precipitation method preparation PLGA microballoon.The precipitation method need for PLGA to be dissolved in miscible with water
Organic solvent (such as DMF and DMSO) obtains PLGA microballoon using the quick diffusion of water-miscible organic solvent in water, however,
Water-miscible organic solvent is difficult to remove completely, and the residual quantity in microballoon is high, is needed through cumbersome, complicated, prolonged processing
Process can just thoroughly remove.
Summary of the invention
Goal of the invention: high and heavy for the residual of surfactant existing for existing emulsion process height, microsphere surface drugloading rate
The high problem of Determination of Residual Organic Solvents existing for the method for shallow lake, it is micro- that the present invention provides a kind of PLGA biodegrade for loading disulfiram drug
The preparation method of ball.
Technical solution: a kind of preparation method for the PLGA biodegradable microspheres loading disulfiram drug of the present invention,
Include the following steps:
(1) poly lactide-glycolide acid (abbreviation PLGA) and disulfiram drug are dissolved in methylene chloride, and by gained
Solution is transferred in syringe;
(2) syringe needle of syringe is submerged in the water being vigorously stirred, water temperature is higher than methylene chloride boiling point, then by double sulphur
Logical sequence/PLGA mixed solution is added drop-wise in water dropwise;
(3) stirring gained lotion, obtains the microballoon of different-grain diameter after being added dropwise to complete, and to thus obtained microsphere classification, takes micron order
Microballoon freeze-drying is to get the PLGA biodegradable microspheres for loading disulfiram drug.
Preferably, in step (1), the intrinsic viscosity of poly lactide-glycolide acid is 0.1~0.5dL/g;Wherein,
The molar ratio of lactic acid and hydroxyacetic acid is 10:1~1:1.Further, poly lactide-glycolide acid and disulfiram drug
Methylene chloride is dissolved according to the mass ratio of 50:1~10:1.
In above-mentioned steps (2), the stirring rate that the water being vigorously stirred generally refers to water is more than or equal to 1000r/min.Into one
Step, water temperature is preferably 40~70 DEG C.Wherein, preferably meet such as between the solvent volume used in the volume and step (1) of water
Lower proportionate relationship: the volume ratio of methylene chloride and water is 1:100~1:10.
Preferably, in step (3), gained lotion is continuously stirred 1~5 hour after being added dropwise to complete, and obtains different-grain diameter
Microballoon, including nanoscale microballoon and micron order microballoon.The mode that membrane filtration thus obtained microsphere can be used divides thus obtained microsphere
Grade, can filter off nanoscale microballoon by filter membrane, micron order microballoon is obtained on filter membrane, and the micron order microballoon is then taken to be freeze-dried
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The utility model has the advantages that the preparation method of the PLGA biodegradable microspheres of load disulfiram drug of the invention combines emulsification
The common advantage and characteristic of method and the precipitation method is dissolved in dichloro by the mixed solution of the PLGA Biodegradable high molecular and disulfiram
It in methane and is transferred in syringe, syringe needle submerges in the water of certain temperature, and disulfiram/PLGA mixed solution is added dropwise
Microballoon is directly formed into water, while low-boiling methylene chloride can quickly volatilize;On the one hand, compared with emulsion process, the present invention
Whole preparation process in stablize emulsion droplet without using surfactant, avoid the surfactant pair of microsphere surface absorption
The influence of drugloading rate;On the other hand, compared with the precipitation method, water-miscible organic solvent is not used in whole preparation process of the invention,
Later period cumbersome removing step is avoided, dissolvent residual is reduced.
Detailed description of the invention
Fig. 1 is the SEM photograph of the PLGA biodegradable microspheres of the load disulfiram drug obtained of embodiment 1.
Specific embodiment
Technical solution of the present invention is described further below with reference to embodiment.
Embodiment 1
By the PLGA Biodegradable high molecular (LA and GA molar ratio are 2:1) and disulfiram medicine that intrinsic viscosity is 0.15dL/g
Object is dissolved in methylene chloride according to the mass ratio of 15:1, and transfers the solution into syringe, and the syringe needle of syringe submerges 45 DEG C of plays
In the distilled water of strong stirring, distilled water stirring rate 1000r/min, under the control of metering pump, by disulfiram/PLGA mixing
Solution is added drop-wise in distilled water dropwise, and the volume ratio of methylene chloride and water is 1:45.It is small that lotion continuously stirred 4 after being added dropwise to complete
Shi Hou, the membrane filtration for being 1 micron with aperture can be obtained load disulfiram drug after taking the microballoon on filter membrane to be freeze-dried
PLGA biodegradable microspheres, SEM picture such as Fig. 1.
Embodiment 2
By the PLGA Biodegradable high molecular (LA and GA molar ratio are 1:1) and disulfiram medicine that intrinsic viscosity is 0.5dL/g
Object is dissolved in methylene chloride according to the mass ratio of 10:1, and transfers the solution into syringe, and the syringe needle of syringe submerges 70 DEG C of plays
In the distilled water of strong stirring, distilled water stirring rate 5000r/min, under the control of metering pump, by disulfiram/PLGA mixing
Solution is added drop-wise in distilled water dropwise, and the volume ratio of methylene chloride and water is 1:10.It is small that lotion continuously stirred 5 after being added dropwise to complete
Shi Hou, the membrane filtration for being 1 micron with aperture can be obtained load disulfiram drug after taking the microballoon on filter membrane to be freeze-dried
PLGA biodegradable microspheres.The SEM figure that gained loads the PLGA biodegradable microspheres of disulfiram drug is close with Fig. 1.
Embodiment 3
By the PLGA Biodegradable high molecular (LA and GA molar ratio are 5:1) and disulfiram medicine that intrinsic viscosity is 0.3dL/g
Object is dissolved in methylene chloride according to the mass ratio of 20:1, and transfers the solution into syringe, and the syringe needle of syringe submerges 50 DEG C of plays
In the distilled water of strong stirring, distilled water stirring rate 3000r/min, under the control of metering pump, by disulfiram/PLGA mixing
Solution is added drop-wise in distilled water dropwise, and the volume ratio of methylene chloride and water is 1:50.It is small that lotion continuously stirred 3 after being added dropwise to complete
Shi Hou, the membrane filtration for being 1 micron with aperture can be obtained load disulfiram drug after taking the microballoon on filter membrane to be freeze-dried
PLGA biodegradable microspheres.The SEM figure that gained loads the PLGA biodegradable microspheres of disulfiram drug is close with Fig. 1.
Embodiment 4
By the PLGA Biodegradable high molecular (LA and GA molar ratio are 3:1) and disulfiram medicine that intrinsic viscosity is 0.4dL/g
Object is dissolved in methylene chloride according to the mass ratio of 30:1, and transfers the solution into syringe, and the syringe needle of syringe submerges 55 DEG C of plays
In the distilled water of strong stirring, distilled water stirring rate 1500r/min, under the control of metering pump, by disulfiram/PLGA mixing
Solution is added drop-wise in distilled water dropwise, and the volume ratio of methylene chloride and water is 1:20.Lotion is continuously stirred 3.5 after being added dropwise to complete
After hour, the membrane filtration for being 1 micron with aperture can be obtained load disulfiram drug after taking the microballoon on filter membrane to be freeze-dried
PLGA biodegradable microspheres.The SEM figure that gained loads the PLGA biodegradable microspheres of disulfiram drug is close with Fig. 1.
Embodiment 5
By the PLGA Biodegradable high molecular (LA and GA molar ratio are 10:1) and disulfiram medicine that intrinsic viscosity is 0.1dL/g
Object is dissolved in methylene chloride according to the mass ratio of 50:1, and transfers the solution into syringe, and the syringe needle of syringe submerges 40 DEG C of plays
In the distilled water of strong stirring, distilled water stirring rate 2000r/min, under the control of metering pump, by disulfiram/PLGA mixing
Solution is added drop-wise in distilled water dropwise, and the volume ratio of methylene chloride and water is 1:100.Lotion is continuously stirred 1 after being added dropwise to complete
After hour, the membrane filtration for being 1 micron with aperture can be obtained load disulfiram drug after taking the microballoon on filter membrane to be freeze-dried
PLGA biodegradable microspheres.The SEM figure that gained loads the PLGA biodegradable microspheres of disulfiram drug is close with Fig. 1.
Comparative example 1
The PLGA biodegradable microspheres of load disulfiram drug are prepared using conventional emulsification method:
By the PLGA Biodegradable high molecular (LA and GA molar ratio are 2:1) and disulfiram medicine that intrinsic viscosity is 0.15dL/g
Object is dissolved in methylene chloride according to the mass ratio of 15:1, is slowly dropped into the water containing 2wt%PVA, the volume ratio of methylene chloride and water
For 1:45.After lotion is continuously stirred 4 hours after being added dropwise to complete, the membrane filtration for being 1 micron with aperture takes the microballoon on filter membrane
It can be obtained the PLGA biodegradable microspheres of load disulfiram drug after freeze-drying.
Comparative example 2
The PLGA biodegradable microspheres of load disulfiram drug are prepared using traditional precipitation method:
By the PLGA Biodegradable high molecular (LA and GA molar ratio are 2:1) and disulfiram medicine that intrinsic viscosity is 0.15dL/g
Object is dissolved in tetrahydrofuran according to the mass ratio of 15:1, disulfiram/PLGA mixed solution is added drop-wise in distilled water dropwise, tetrahydro
The volume ratio of furans and water is 1:45.After solution is continuously stirred 4 hours after being added dropwise to complete, the filter membrane mistake for being 1 micron with aperture
Filter can be obtained the PLGA biodegradable microspheres of load disulfiram drug after taking the microballoon on filter membrane to be freeze-dried.
The solvent on the PLGA biodegradable microspheres surface of measurement Examples 1 to 5 load disulfiram drug obtained is residual respectively
Allowance and the carrying drug ratio for being adsorbed on microsphere surface load disulfiram with made from 1~2 conventional emulsification method of comparative example and the precipitation method
The PLGA biodegradable microspheres of drug are compared, as a result such as the following table 1.
1 conventional emulsification method of table and the precipitation method with it is disclosed by the invention prepare micro-sphere method result compared with