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Nanocarrier and preparation method for co-delivery of drugs and genes based on oligonucleotides

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 of loaded drugs, and achieve the improvement of anti-tumor effect, Reduce toxic side effects and improve accumulation effect

Active Publication Date: 2018-05-25
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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  • Nanocarrier and preparation method for co-delivery of drugs and genes based on oligonucleotides
  • Nanocarrier and preparation method for co-delivery of drugs and genes based on oligonucleotides
  • Nanocarrier and preparation method for co-delivery of drugs and genes based on oligonucleotides

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'-GGGAGGACGAUGCGGAUCAGCCAUGUUUACGUCACUCCUUGUCAAUCCUCAUCGGC-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 a nano-carrier based on oligonucleotides for co-delivery of drugs and genes and a preparation method thereof. The oligonucleotide solution is mixed with the drug solution, and after being placed at room temperature for 1-100 minutes, it is mixed with siRNA to obtain a mixed solution; Add the cationic polymer solution to the mixed solution obtained in step 1), and incubate at room temperature for 1 to 100 minutes to obtain the polycation complex that co-loads the drug and the gene; add the anionic polymer solution to the polycation complex obtained in step 2), Place and incubate at room temperature for 1-100 min to obtain the nanocarrier based on oligonucleotides for co-delivery of drugs and genes. The preparation process of the oligonucleotide-based nanocarrier for co-delivery of drugs and genes prepared by the invention is simple and feasible, and can be used for the treatment of various 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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Patent Type & Authority Patents(China)
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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