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Immunostimulatory composition and use thereof

A technology of immune stimulation and composition, which is applied in the field of biopharmaceuticals, can solve the problems of huge difference in effect, and achieve the effect of excellent effect, high antibody level, and enhanced immune effect

Pending Publication Date: 2021-06-18
JIANGSU THERAVAC BIO PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the structural diversity of CpG oligodeoxynucleotides, the effects of CpG adjuvants of different sequences vary greatly

Method used

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  • Immunostimulatory composition and use thereof
  • Immunostimulatory composition and use thereof
  • Immunostimulatory composition and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Embodiment 1 prepares immunostimulatory composition of the present invention and hepatitis B vaccine

[0073] 1. HBsAg stock solution: the amino acid sequence of HBsAg protein is shown in SEQ ID NO:1.

[0074] The HBsAg protein is prepared by HBsAg gene recombinant yeast cells, and the yeast cell types include Hansenula, Saccharomyces and Pichia, preferably Hansenula. For the specific preparation steps, refer to the Chinese patent application CN108330145A, the HBsAg gene recombinant Hansenula cells are fermented and cultured, and the cells are harvested. Purified by bacteria-breaking treatment, silica gel adsorption, column chromatography and TFF.

[0075] 2. HBcAg stock solution: the amino acid sequence of HBcAg protein is shown in SEQ ID NO:2.

[0076] The HBcAg protein is prepared by HBcAg gene recombinant yeast cells, and the yeast cell types include Hansenula, Saccharomyces cerevisiae and Pichia pastoris, preferably Hansenula. For the specific preparation step...

Embodiment 2

[0084] Example 2 Screening experiment of CPG oligodeoxynucleotides

[0085] 1. Experimental animals: C57BL / 6(N) mice, male, 4 weeks old, 135 mice, Shanghai Lingchang Experimental Animal Technology Co., Ltd.

[0086] 2. Experimental grouping: see Table 2, each injection volume is 100 μL / rat. Among them, group A is the negative control, and the PBS solution is 100 μL per mouse.

[0087] Table 2 Grouping of experimental animals

[0088]

[0089] 3. Experimental steps: Take the spleen on the 7th day after the mouse is immunized, and prepare spleen lymphocytes according to the conventional method, as follows: take the spleen aseptically: cut the spleen with sterile forceps and scissors, put it in a 70 μm cell strainer, Place in a plate containing 2ml of pre-cooled 2% FBS (purchased from GIBCO)-PBS; grind the spleen with a grinding rod, and the spleen cells enter the plate through a sieve to obtain a cell suspension, and put the suspension into the plate with a Pasteur pipett...

Embodiment 3

[0092] The screening experiment of embodiment 3 immunostimulatory composition

[0093] 1. Experimental animals: C57BL / 6(N) mice, male, 4 weeks old, 81, Shanghai Lingchang Experimental Animal Technology Co., Ltd.,

[0094] 2. Experimental grouping: see Table 3, each injection volume is 100 μL / rat. Among them, group A is the negative control, and the PBS solution is 100 μL per mouse.

[0095] Table 3 Grouping of experimental animals

[0096]

[0097]

[0098] 3. Experimental steps: the same as in Example 2.

[0099] 4. Experimental results: ELISPOT spot results see image 3 and Figure 4 , the results showed that the combined application of CpG T1~T3 and QS21 had a highly efficient synergistic effect, and the level of HBsAg and HBcAg-specific IFN-γ induced was significantly higher than that of other CpG adjuvants such as CpG1018 and CpG7909, which had unexpected immune effects .

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Abstract

The invention provides an immunostimulatory composition, the immunostimulatory composition comprises saponin and CpG oligodeoxynucleotide, or the immunostimulatory composition is composed of an adjuvant containing the saponin, and the CpG oligodeoxynucleotide, wherein the CpG oligodeoxynucleotide sequence is provided with two or more copies of a 5'-TTCGTT-3' sequence motif or a 5'-TCGTCGTCG-3 'sequence motif. The invention provides the use of the immunostimulatory composition in the preparation of a medicament for the treatment of diseases.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals. Specifically, the present invention relates to an immunostimulatory composition comprising saponin and CpG oligodeoxynucleotides, or the immunostimulatory composition is composed of an adjuvant comprising saponin and CpG oligodeoxynucleosides acid composition. Wherein, the CpG oligodeoxynucleotide sequence has two or more copies of the 5'-TTCGTT-3' motif or the 5'-TCGTCGTCG-3' motif. The invention also relates to the pharmaceutical use of said immunostimulatory composition. Background technique [0002] CpG oligodeoxynucleotides are a new class of immunostimulants discovered in recent years. Their chemical essence is deoxyoligonucleotides containing cytosine guanine dinucleotides, which have similar immunity to natural CpG pattern recognition receptors. It can bind to Toll-like receptors on the cell membrane and effectively trigger mammalian immune responses through the TLR9 signaling pathway...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/39A61K39/00A61P31/00A61P33/00A61P35/00A61P37/04
CPCA61K39/39A61K39/00A61P31/00A61P33/00A61P35/00A61P37/04A61K2039/55561A61K2039/55577Y02A50/30A61P31/20A61K39/12A61K2039/572A61K2039/575A61K2039/70C12N2710/16734A61P31/12A61P31/04A61K39/245A61K39/29A61P31/22A61K31/711C07K16/2803A61K39/292
Inventor 葛君李建强孙娇娇周童任苏林顾月黄红颖王世伟黄精俸
Owner JIANGSU THERAVAC BIO PHARMA CO LTD
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