Composite nano-carrier capable of loading nucleic acid drugs efficiently and preparation method of composite nano-carrier

A nano-carrier and nucleic acid technology, which is applied in the direction of medical preparations of non-active ingredients, pharmaceutical formulas, non-effective ingredients of polymer compounds, etc., can solve the problems of inability to reach the target site, increased thrombus formation, low loading efficiency, etc., to achieve guaranteed In vivo compatibility, easy removal, low cost effect

Active Publication Date: 2013-01-09
ARMY MEDICAL UNIV
3 Cites 2 Cited by

AI-Extracted Technical Summary

Problems solved by technology

Cationic polymer carrier, because it complexes with nucleic acid drugs mainly through electrostatic interaction, the final complex is easily decomplexed by the polyanions that exist in large quantities in blood and extracellular fluid, so that it cannot reach the target site to achieve ideal transfection
In addition, for low-molecular-weight nucleic acid drugs (such as antisense nucleotides, siRNA and microRNA), their rigid structure and low charge density often lead to incomplete aggr...
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Method used

The present invention takes degradable organic material as main carrier material to design the composite nano-carrier that can efficiently load nucleic acid class drug, improves composite nano-carrier to antisense nucleotide, siRNA and antisense nucleotide, siRNA and The loading or packaging efficiency of microRNA, etc., promotes the escape of endosomes/lysosomes; on the other hand, it will not increase the cytotoxicity or tissue damage of the carrier system. Different from the conventional double-emulsion method used in previous studies, in which the complex aqueous solution formed by polyamines of different molecular weights and nucleic acid drugs is used as the internal water phase, the method designed in the present invention uses nucleic acid drugs as the internal water phase, and the Polyamines of different molecular weights are dissolved in the o...
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Abstract

The invention relates to a composite nano-carrier capable of loading nucleic acid drugs efficiently and a preparation method of the composite nano-carrier. The composite nano-carrier is composed of a degradable organic material and polyamine with different molecular weights, wherein the mass ratio of the polyamine with different molecular weights to the degradable organic material is (0.005: 100)-(3:7), the composite nano-carrier is spherical, the particle size of the composite nano-carrier ranges from 80 nm to 950 nm, and the nucleic acid drugs are distributed in the composite nano-carrier uniformly. The preparation method of the composite nano-carrier includes steps of firstly, dissolving the nucleic acid drugs into water to obtain internal water phase, dissolving the degradable organic material and the polyamine with different molecular weights into organic solvent to obtain organic phase, taking polyving alcohol aqueous solution as external water phase; secondly, emulsifying the internal water phase into the organic phase under ultrasonic action of a probe so as to form water-in-oil colostrum, emulsifying the water-in-oil colostrum into the external water phase under ultrasonic action of the probe so as to obtain oil-in-water multiple emulsion, stirring the multiple emulsion by magnetic force at room temperature so as to remove the organic solvent in the organic phase by volatilizing, performing centrifugal separation after nano-particles are solidified, washing the nano-particles with double distilled water so as to obtain the composite nano-carrier capable of loading nucleic acid drugs.

Application Domain

Genetic material ingredientsMacromolecular non-active ingredients +1

Technology Topic

Water in oilPolyvinyl alcohol +13

Image

  • Composite nano-carrier capable of loading nucleic acid drugs efficiently and preparation method of composite nano-carrier
  • Composite nano-carrier capable of loading nucleic acid drugs efficiently and preparation method of composite nano-carrier
  • Composite nano-carrier capable of loading nucleic acid drugs efficiently and preparation method of composite nano-carrier

Examples

  • Experimental program(20)

Example Embodiment

[0033] Example 1
[0034] First, 1 nmol of pDNA encoding β-galactosidase was dissolved in 100 μl of water to obtain the internal aqueous phase; 50 mg of polylactic acid (PLA, molecular weight 15 kDa) and 0.5 mg of branched polyethyleneimine with molecular weight of 800 Da were dissolved in 1 The organic phase is obtained in ml of dichloromethane; then the internal water phase is emulsified in the organic phase under the ultrasound of the probe to form water-in-oil colostrum, which is emulsified in 10 ml of 0.5% by mass under ultrasound of the probe A water-in-oil-in-water double emulsion is obtained in an aqueous solution of polyvinyl alcohol (molecular weight 8.8 kDa, degree of hydrolysis 88%); the resulting double emulsion is magnetically stirred at room temperature to volatilize and remove the organic solvent from the organic phase. After the nanoparticles are solidified, they are centrifuged. After washing with double distilled water, a composite nanocarrier loaded with pDNA can be obtained, and the loading efficiency of pDNA is 92%.

Example Embodiment

[0035] Example 2
[0036] First, 0.5 nmol Bcl-2 antisense oligonucleotide was dissolved in 100 μl of water to obtain the internal aqueous phase; 50 mg of polylactide-glycolide (PLGA, lactic acid/glycolic acid molar ratio 50:50, molecular weight 20 kDa) and 2.5 mg of branched polyethyleneimine with a molecular weight of 1.8 kDa was dissolved in 1 ml of dichloromethane to obtain the organic phase; then the internal water phase was emulsified in the organic phase under the ultrasonic action of the probe to form water-in-oil colostrum. The probe was emulsified in 20 ml of 1% polyvinyl alcohol (molecular weight 8.8 kDa, hydrolysis degree 88%) aqueous solution under ultrasound to obtain a water-in-oil-in-water double emulsion; the resulting double emulsion was removed by magnetic stirring at room temperature The organic solvent of the organic phase, after the nanoparticles are solidified, centrifuged and washed with double distilled water, the composite nanocarrier loaded with Bcl-2 antisense oligonucleotides can be obtained, and the loading efficiency of the oligonucleotides is 95% .

Example Embodiment

[0037] Example 3
[0038] First, 1 nmol Bcl-2 antisense oligonucleotide was dissolved in 150 μl water to obtain the internal aqueous phase; 50 mg polylactide-glycolide (PLGA, lactic acid/glycolic acid molar ratio 75:25, molecular weight 25 kDa) and 5 mg of branched polyethyleneimine with a molecular weight of 10 kDa was dissolved in 1 ml of chloroform to obtain an organic phase; then the internal water phase was emulsified in the organic phase under ultrasound of the probe to form water-in-oil colostrum, which was used in the ultrasound of the probe Under emulsification in 50 ml of 0.8% polyvinyl alcohol (molecular weight 25 kDa, hydrolysis degree 80%) aqueous solution to obtain a water-in-oil-in-water double emulsion; the resulting double emulsion is magnetically stirred at room temperature to volatilize and remove the organic phase After the nanoparticles are solidified, they are centrifuged and washed with double distilled water to obtain composite nanocarriers loaded with Bcl-2 antisense oligonucleotides. The loading efficiency of the oligonucleotides is 94%.
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PUM

PropertyMeasurementUnit
Particle size80.0 ~ 950.0nm
Molecular weight1.0 ~ 100.0
Molecular weight15.0
tensileMPa
Particle sizePa
strength10

Description & Claims & Application Information

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