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Oligonucleotide-based nano carrier for co-delivering drug and gene and preparation method of nano carrier

An oligonucleotide and nanocarrier technology is applied in the field of nanocarriers and preparation of co-delivery drugs and genes based on oligonucleotides, which can solve the problems of low drug loading and improve the anti-tumor effect. Reduce toxic side effects and improve accumulation effect

Active Publication Date: 2015-01-07
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the simple aptamer loading drug has a low drug loading capacity. In order to improve the drug loading ability, Sanyong Jon's research group modified the end of the aptamer A10

Method used

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  • Oligonucleotide-based nano carrier for co-delivering drug and gene and preparation method of nano carrier
  • Oligonucleotide-based nano carrier for co-delivering drug and gene and preparation method of nano carrier
  • Oligonucleotide-based nano carrier for co-delivering drug and gene and preparation method of nano carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The oligonucleotides used in this example are formed by the annealing of two complementary single-stranded nucleotides consisting of (CGA) repeat sequences (CGA) 7 , strand 1: 5'-CGACGACGACGACGACGACGA-3'; strand 2: 5'-TCGTCGTCGTCGTCGTCGTCG-3', as shown in SEQ ID NO.1, SEQ ID NO.2; siRNA specifically targets vascular endothelial growth factor (VEGF ) siRNA, the sequence is: sense strand 5'-ACAUCACCAUGCAGAUUAUdTdT-3'; antisense strand 5'-dTdTUGUAGUGGUACGUCUAAUA-3' as shown in SEQ ID NO.3 and SEQ ID NO.4.

[0037] Mix 100 μL of doxorubicin aqueous solution with a concentration of 50 μM and 120 μL of an oligonucleotide solution with a concentration of 10 μM, vortex for 20 s, and after standing at room temperature for 5 min, add specific For siRNA targeting VEGF, vortex for 20s, and mix well to obtain a mixed solution.

[0038] Add 200 μL of the above-prepared mixed solution drop by drop into the polyetherimide (PEI) solution with a volume of 200 μL and a concentration of 8...

Embodiment 2

[0041] The aptamer used in this example is aptamer A10, the sequence is: 5'-GGGAGGACGAUGCGGAUCAGC CAUGUUUACGUCACUCCUUGUCAAUCCUCAUCGGC-3', as shown in SEQ ID NO.5; siRNA is siRNA specifically targeting VEGF, and the sequence is as shown in SEQ ID NO.3, shown in SEQ ID NO.4.

[0042] Mix 100 μL of doxorubicin aqueous solution with a concentration of 20 μM and 80 μL of aptamer A10 with a concentration of 10 μM, vortex for 20 s, and after standing at room temperature for 5 min, add a specific target with a mass ratio of 1:2 to doxorubicin. Add siRNA to VEGF, vortex for 20s, mix well, take 200μL of the mixed solution drop by drop into the PLL solution with a concentration of 200μg / mL and a volume of 150μL under the vortex condition, continue to vortex for 20s, and incubate at room temperature for 30min. A polycation complex co-loaded with doxorubicin and siRNA. The polycation complex co-loaded with doxorubicin and siRNA prepared above has a particle size of 107.9 nm, a polydispersi...

Embodiment 3

[0045] The oligonucleotides used in this example are formed by the annealing of two complementary single-stranded nucleotides consisting of (CGA) repeat sequences (CGA) 5 , strand 1: 5'-CGACGACGACGACGA-3'; strand 2: 5'-TCGTCGTCGTCGTCG-3', the sequence is shown in SEQ ID NO.6 and SEQ ID NO.7, and siRNA specifically targets multidrug resistance genes The sequence of siRNA for MDR-1 is: positive sense strand: 5'-GGAUAUUAGGACCAUAAAUdTdT-3', antisense strand 5'-AUUUAUGGUCCUAAUAUCCdTdT-3', as shown in SEQ ID NO.8 and SEQ ID NO.9.

[0046] Mix 100 μL daunorubicin aqueous solution with a concentration of 40 μM and 100 μL oligonucleotide solution with a concentration of 10 μM, vortex for 20 s, and after standing at room temperature for 5 min, add specific For siRNA targeting multidrug resistance gene 1 (MDR-1), vortex for 20s, mix well, add 200 μL of the above mixed solution drop by drop into the PPI solution with a concentration of 100 μg / mL and a volume of 300 μL under vortex conditi...

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Abstract

The invention discloses an oligonucleotide-based nano carrier for co-delivering drug and gene and a preparation method of the nano carrier. The method comprises the following steps: 1, mixing an oligonucleotide solution with a drug solution, standing at room temperature for 1-100 minutes, and then mixing with siRNA to obtain a mixed solution; 2, adding a cationic polymer solution to the mixed solution obtained in the step 1), standing and incubating at room temperature for 1-100 minutes, so as to obtain a polycation compound co-loading drug and gene; 3, adding an anionic polymer solution to the polycation compound obtained in the step 2), standing and incubating at room temperature for 1-100 minutes, so as to obtain an oligonucleotide-based nano carrier for co-delivering drug and gene. The preparation process of the oligonucleotide-based nano carrier for co-delivering drug and gene is simple and feasible and the oligonucleotide-based nano carrier for co-delivering drug and gene can be applied to treatment of a plurality of cancers such as liver cancer, lung cancer, breast cancer and ovarian cancer.

Description

technical field [0001] The invention belongs to the field of pharmaceutical preparations, and relates to a co-delivery drug delivery system, in particular to a nano-carrier and a preparation method for co-delivering drugs and genes based on oligonucleotides. Background technique [0002] Cancer is a common and frequently-occurring disease that seriously threatens human health. The global mortality rate is as high as 13%, and the incidence rate is increasing year by year. Therefore, the task of cancer prevention and treatment is very arduous. There are three main methods of cancer treatment: drug therapy, gene therapy and radiation therapy. Among them, drug therapy, also known as chemotherapy, is currently a widely used treatment method in clinical practice. Most of the antitumor drugs used in it can directly and quickly kill tumor cells. However, the non-specificity of antitumor drugs also enhances the effects of chemotherapy. Nausea, vomiting, hair loss and other toxic sid...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K47/36A61K47/34A61K47/32A61K47/18A61K45/00A61K31/704A61P35/00
Inventor 张娜刘婷先刘永军刘春喜王明芳
Owner SHANDONG UNIV
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