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Antisense oligodeoxynucleotide and application thereof

An antisense oligonucleotide and immune adjuvant technology, which can be applied to medical preparations containing active ingredients, biochemical equipment and methods, bacterial antigen components, etc., can solve the problem of no antisense oligonucleotides, etc. To achieve the effect of enhancing the inoculation effect, improving the application effect, and easy to adjust the adjuvant concentration

Pending Publication Date: 2017-01-18
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Antisense oligonucleotides can be artificially synthesized in large quantities. At present, there is no application of antisense oligonucleotides as immune adjuvants

Method used

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  • Antisense oligodeoxynucleotide and application thereof
  • Antisense oligodeoxynucleotide and application thereof
  • Antisense oligodeoxynucleotide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Embodiment 1: Effect test of immune adjuvant

[0061] Experimental method: select inbred strain Balb / c female mice without specific pathogens, aged 6 to 8 weeks, with a weight of 17g±0.5g, 30 were randomly divided into 5 groups, according to body weight, 10 μL of pentobar per gram Bital sodium (10mg·mL -1 ) into the anesthetized mice by intraperitoneal injection, and injected the vaccine between the skins of the thighs of the mice in each group for a total of three inoculations with a time interval of three weeks. After the first vaccination, blood was collected from the mice one day before each vaccination, and the mice were sacrificed two weeks after the last vaccination and blood was collected to prepare cell-free serum and detect the titer of antigen-specific IgG by ELISA. The formulations of vaccines and adjuvants used in each group are shown in Table 1.

[0062] Table 1 Grouping and formulation of vaccine adjuvant dosage forms

[0063]

[0064] The IgG titer...

Embodiment 2

[0074] Embodiment 2: dose-dependent test

[0075] Test method: Select inbred strain Balb / c female mice without specific pathogens, aged 6-8 weeks, with a weight of 17g±0.5g, 36 were randomly divided into 6 groups, and the inoculation method and the determination of antibody IgG titer The method is the same as above. The vaccine adjuvant concentration groups and formulations of each group are shown in Table 2 below.

[0076] Table 2 Concentration grouping and formula of ASO adjuvant

[0077]

[0078]

[0079] The results of dose-dependent animal experiments such as image 3 As shown, when the dose of ASO was 5 μg per mouse, the highest antibody titer was produced, and the final concentration of ASO dose was 0.25 μg g -1 , that is, 0.25 μg g -1 In order to enhance the optimal adjuvant dose of vaccine immunogenicity, continuing to increase the adjuvant concentration cannot further improve the immune protection effect of the vaccine.

Embodiment 3

[0080] Embodiment 3: Cytotoxicity test of ASO

[0081] Mouse BMDC and human HeLa cell lines were selected for culture, and different concentrations of ASO, ASO control and PEI-B25 (a cationic polymer newly confirmed to have the function of an immune adjuvant, which has relatively low cytotoxicity) were added to the cell culture fluid. Strong, as a positive control group), cultured for 24h, 48h, and 72h, respectively, and carried out CCK-8 experiment to detect the cell survival rate of ASO, ASO control and PEI groups.

[0082] Experimental results such as Figure 4 with Figure 5 As indicated, the adjuvant concentration was 0.6 μg·mL -1 When , PEI, ASO, and ASO control did not show obvious toxicity, and the concentration was 4.8 μg·mL -1 , PEI began to exhibit cytotoxicity, and the cell survival rate began to decrease, and with the increase of the concentration, the stronger the toxicity of PEI, the lower the cell survival rate. However, the ASO and ASO control groups did n...

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Abstract

The invention provides an antisense oligodeoxynucleotide, and the serial number is 5' AGGTCCTGGAGTCCAGCA 3'. The antisense oligodeoxynucleotide has the beneficial advantages that as an immunologic adjuvant, the antisense oligodeoxynucleotide adopts immunomodulatory mechanism different from prior adjuvants, and sensitivity of the response of the immune systems is increased through temporarily suppressing the expression of cytokines interleukin 10 on part of vaccine delivery, thus promoting inoculation effect of the vaccine. Besides, as a safe, stable, efficient, and inexpensive immunologic adjuvant with water solubility, the antisense oligodeoxynucleotide can be used as multiple channels of vaccine delivery, can easily adjust the concentration, and can be used through the concerted application, and is capable of changing application effects of prior immunologic adjuvants; meanwhile, through joint utilization with other adjuvants, the dose of thenecessary adjuvants with large side effect is lowered, hereby optimizing the functions of the adjuvants.

Description

technical field [0001] The invention relates to the field of immunology, in particular to an antisense oligonucleotide and its application. Background technique [0002] An immune adjuvant is a non-specific immune enhancer that can effectively enhance the intensity of the immune response or change the type of immune response when it is injected into the body together with the antigen or in advance. [0003] An ideal vaccine immune adjuvant can enhance the immunogenicity of vaccine antigens, prolong the response time, or change the type of immune response, and can induce a more comprehensive and durable immune response in immunocompromised individuals (the elderly, children, etc.). The use of immune adjuvants can reduce the dose of antigen required for effective vaccination, thereby reducing costs and making limited vaccine antigens benefit more people. However, the types of adjuvants currently approved for clinical use are very limited. Among them, aluminum salt adjuvant i...

Claims

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Application Information

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IPC IPC(8): C12N15/113A61K39/39A61K39/08A61K39/05A61K39/29A61P31/04A61P31/20
CPCA61K39/05A61K39/08A61K39/12A61K39/39C12N15/1136A61K2039/55561C12N2320/31C12N2320/53C12N2310/11C12N2730/10034A61K2300/00
Inventor 丁智张小影刘宁华董磊张峻峰
Owner NANJING UNIV
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