Cpg Single Strand Deoxynucleotides for Use as Adjuvant

a single-strand deoxynucleotide and adjuvant technology, applied in the field of single-strand deoxynucleotide containing cpg dinucleotides, can solve the problems of not thoroughly eliminating latent hbvs inside the infected cells, no effective treatment of rabies infections, and severe effects on the health of people around the globe, so as to improve the immune response of the body, enhance the immunogenicity of the hbv vaccin

Inactive Publication Date: 2008-01-03
CHANGCHUN HUAPU BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0024] As examples, the combination use of a rabies vaccine with the adjuvant of the present invention not only significantly improved the immune response of the body to the vaccine, but also reduced the total administrations needed for the immunization; when CpG ODN is combined with HbsAg, it enhanced the immu...

Problems solved by technology

Currently, there is no effective treatment for rabies infections.
Hepatitis B is a liver disease caused by hepatitis B virus (HBV), which has severely affected the health of people around the globe.
Although the anti...

Method used

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  • Cpg Single Strand Deoxynucleotides for Use as Adjuvant
  • Cpg Single Strand Deoxynucleotides for Use as Adjuvant
  • Cpg Single Strand Deoxynucleotides for Use as Adjuvant

Examples

Experimental program
Comparison scheme
Effect test

example 1

Design of the Single Strand Deoxynucleotide Containing CpG

[0041] The sequences were designed as follows:

(1) (G)n(L)n X1X2CGY1Y2(M)n (G)nX1 = A, T, G; X2 = A, T; Y1 = A, T; Y2 = A, T, C; L, M = A, T,C, G; n is 0-6.5′-ggggTCgTTCgTCgTTgggggg-3′(SEQ ID NO: 1)[121]5′-ggggATAACgTTgCgggggg-3′(SEQ ID NO: 2)[143]5′-ggggTgCAACgTTCAgggggg-3′(SEQ ID NO: 3)[402]5′-ggggTCCTACgTAggAgggggg-3′(SEQ ID NO: 4)[123]5′-ggggTCCATgACgTTCCTgAAgggggg-3′(SEQ ID NO: 5)[603]5′-gggggACgTCgCCggggggg-3′(SEQ ID NO: 6)[118]5′-ggATCCgTACgCATgggggg-3′(SEQ ID NO: 7)[320]5′-gggggAATCgATTCgggggg-3′(SEQ ID NO: 8)[154]5′-gggATgCATCgATgCATCgggggg-3′(SEQ ID NO: 9)[464]5′-ggTgCgACgTCgCAgggggg-3′(SEQ ID NO: 10)[471]5′-gggACgTACgTCgggggg-3′(SEQ ID NO: 11)[390]5′-gggggATCgACgTCgATCgggggg-3′(SEQ ID NO: 12)[322]5′-ggCgATCgATCgATCggggggg-3′(SEQ ID NO: 13)[333]5′-ggggTCgATCgATCgAgggggg-3′(SEQ ID NO: 14)[113]5′-ggTCgCgATCgCgAgggggg-3′(SEQ ID NO: 15)[307]5′-ggGGTCAACGTTGAgggggG-3′(SEQ ID NO: 16)[156]5′-gTCgTTTTCgTCgACgAATTgggggggg-...

example 2

Synthesis of the Single Strand Deoxynucleotide Containing CpG

[0044] DNA fragment was synthesized by solid phasephosphoramidite triestermethod. The method has been widely used in chemical synthesis of DNA due to its advantages of fast, high efficiency, etc.

[0045] Chemical synthesis of DNA is different from enzymatic synthesis of DNA. The latter is synthesized from 5′ end to 3′ end, while the former starts from 3′ end. The synthesis steps are as follows:

[0046] 1. De-Protection

[0047] Trichloroacetic acid was used to remove protective group DMT conjugated to the nucleotides on Controlled Pore Glass (CPG) to obtain free 5′ hydroxyl ends for being used in the following condensation reaction.

[0048] 2. Activation

[0049] The nucleotides units protected by phosphoramidite was mixed with tetrazolium activator and then loaded into a synthesis column to form an active intermediate of phosphoramiditetetrazole (the 3′ end of which is activated, while the 5′ end of which is still under DMT pro...

example 3

Effects of Different Dosages of a CpG ODN on Antibody Production Stimulated by Rabies Vaccine

[0059] 1. Experimental animals: 200 white mice, with half males and half females, weighted from 18 to 22 g, aged from 6 to 8 weeks, and purchased from Beijing Weitonglihua Experimental Animal Ltd.

[0060] 2. Rabies vaccine: 1 ml / vial (containing 2.5 IU), purchased from Changchun Institute of Biological Product.

[0061] 3. CpGODN: synthesized by Shanghai Shenggong Biotechnology Service Ltd.

[0062] 4. Experimental groups: 8 mice for each group, with half males and half females.

[0063] rabies vaccine

[0064] rabies vaccine+1.25 μg CpG667

[0065] rabies vaccine+1.25 μg CpG705

[0066] rabies vaccine+1.25 μg CpG309

[0067] rabies vaccine+1.25 μg CpG205

[0068] rabies vaccine+5 μg CpG667

[0069] rabies vaccine+5 μg CpG705

[0070] rabies vaccine+5 μg CpG309

[0071] rabies vaccine+5 μg CpG205

[0072] rabies vaccine+20 μg CpG667

[0073] rabies vaccine+20 μg CpG705

[0074] rabies vaccine+20 μg CpG309

[0075] rabi...

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Abstract

The present invention provides an adjuvant, which includes at least one single strand deoxynucleotide containing a CpG dinucleotide. The single strand deoxynucleotide comprises one or more CpG dinucleotides. When used in combination with rabies vaccine, HBV vaccine or other vaccines, the adjuvant can significantly improve the immune effect of the vaccine.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a single strand deoxynucleotide containing CpG dinucleotides, and particularly relates to a single strand deoxynucleotide containing CpG dinucleotides as an adjuvant in hepatitis B virus (HBV) vaccine or rabies vaccine. The present invention further relates to sequences of the single strand deoxynucleotide containing CpG dinucleotides. BACKGROUND OF THE INVENTION [0002] CpG ODN is a type of oligodeoxynucleotides centered with non-methylated cytosine and guanine nucleotide (CpG). Typically, the CpG is flanked by base sequences in the following manner: 5′PurPurCGPyrPyr 3′, i.e., 2 purines at its 5's end and 2 pyrimidines at its 3′ end. (G Mutwiri, R. Pontarollo, S. BaBIUK. Biological activity of immunostimulatory CpG ODN motif in domestic animals. Veterinary Immunopathology, 2003, 91: 89-103). Studies indicate CpG ODN can activate a variety of immune effector cells, in which non-methylated CpG dinucleotides is believed to ...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K31/7088A61K45/00A61P31/12A61P31/18C07H21/00C12N15/117
CPCA61K39/39C12N2310/16C12N15/117A61K2039/55561A61P1/16A61P31/12A61P31/18A61P31/20A61P37/02A61P43/00
Inventor WANG, LIYINGBAO, MUSHENGYU, YONGLI
Owner CHANGCHUN HUAPU BIOTECHNOLOGY CO LTD
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