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Vectors for the treatment of Alzheimer's disease

A carrier and carrier technology, applied in gene therapy, virus/phage, medical raw materials derived from virus/phage, etc., can solve the problems of unavailable vaccine technology and insufficient effect, so as to improve nursing care and improve the quality of life , the effect of reducing medical expenses

Inactive Publication Date: 2011-12-07
DNAVEC CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its effect is still insufficient
[0009] As mentioned above, although vaccine therapy itself is recognized as effective, there is no example of vaccine technology that can provide a safer and more effective vaccine, and the current situation is that its development is still strongly desired

Method used

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  • Vectors for the treatment of Alzheimer's disease
  • Vectors for the treatment of Alzheimer's disease
  • Vectors for the treatment of Alzheimer's disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0204] [Example 1] Construction of SeV vector carrying Aβ42 gene

[0205] (1) Construction of the Not I fragment of the Aβ42 gene

[0206] For the Aβ42 gene, assembly was performed by PCR using multiple primers covering the entire length of the human amyloid β peptide sequence (1-42) (SEQ ID NO: 1). The nucleotide sequence of Aβ42 was optimized considering human codon usage. The obtained sequence has the following structure: the Igκ secretion signal is bound to the N-terminal side, and the transcription signal of Sendai virus is added to the C-terminal side ( figure 1 , SEQ ID NO: 2).

[0207] Build methods such as figure 2 shown. First, six long primers F1 (SEQ ID NO: 4), F2 (SEQ ID NO: 5), R1 (SEQ ID NO: 6), R2 (SEQ ID NO: 7 ), R3 (SEQ ID NO: 8), R4 (SEQ ID NO: 9) mixed. Using this primer compound, PCR was performed without adding a template. Then, using the PCR product as a template, PCR was further performed using two kinds of primers F1-1 (SEQ ID NO: 10) and R4-1 ...

Embodiment 2

[0211] [Example 2] Construction of a SeV vector carrying a fusion gene of Aβ42 and CTB (CTB-Aβ42)

[0212] (1) Construction of NotI fragment of CTB-Aβ42 gene

[0213] The NotI fragment of CTB-Aβ42 has the following structure: the N-terminal side of the sequence of the human amyloid β sequence (1-42) is linked to the cholera toxin B subunit sequence (SEQ ID NO: 14 ) connection, and the transcription signal of Sendai virus was added at its C-terminal side ( image 3 , SEQ ID NO: 15). Moreover, in order to increase expression efficiency, the nucleotide sequences of CTB and Aβ were changed according to human codon usage, but the amino acid sequences were not changed.

[0214] The gene was constructed by synthesizing the full length of the gene by performing PCR using multiple long primers covering the full length of the gene. Specifically, 8 kinds of long primers [CTB-AβF-1 (SEQ ID NO: 17), F-2 (SEQ ID NO: 18), F-3 (SEQ ID NO : 19), F-4 (SEQ ID NO: 20), R-1 (SEQ ID NO: 21), R-...

Embodiment 3

[0217] [Example 3] Construction of the SeV vector carrying the fusion gene of Aβ42 and IL-4

[0218] (1) Construction of the NotI fragment of the fusion gene of Aβ42 and IL-4

[0219] The fusion of Aβ42 gene and mouse IL-4 was carried out by partially overlapping and assembled by PCR.

[0220] Aβ42 gene utilization contains Aβ42 EcoRI fragment (embodiment 1: figure 2 ) plasmid preparation. Meanwhile, mouse IL-4 gene (SEQ ID NO: 27) was prepared as cDNA by the following procedure. mRNA was extracted from the spleen of mice (BALB / cA), reverse-transcribed using IL-4-specific primers, amplified by PCR, and subcloned into cloning plasmids. The resulting plasmid with mouse IL-4 cDNA as an insert was used for construction.

[0221]Specifically, using the mouse IL-4 plasmid as a template, two primers NotI-IL4-F (SEQ ID NO: 29) and IL4-R (SEQ ID NO: 30) were used for PCR. On the other hand, using the Aβ42 plasmid as a template, PCR was performed with primers Aβ42-F (SEQ ID NO: 31...

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Abstract

The present invention provides methods for efficiently inducing anti-Aβ antibody and methods for preventing and treating Alzheimer's disease. The present inventors successfully induced anti-Aβ antibody in a highly efficient manner by administering an RNA viral vector that expresses a fusion protein between an AB5 toxin B subunit and an Aβ antigen peptide. Administration of the vector resulted in a significant increase of anti-Aβ antibody in plasma, and decrease in the Aβ level in brain tissues and decrease in the anti-Aβ antibody-positive area. The present invention enables more efficient vaccine gene therapy for preventing and treating Alzheimer's disease.

Description

technical field [0001] The present invention relates to a novel gene transfer vector for efficiently inducing anti-Aβ antibody and preventing and treating Alzheimer's disease, a vaccine for active immunization containing the vector, and the like. Background technique [0002] With the entry into the so-called aging society, the number of patients with neurodegenerative diseases is gradually increasing. For example, the number of people in Japan is 1 million, and the number of people in the United States is 4.5 million. It is estimated that there are more than 15 million Alzheimer's disease patients in the world, and it is expected that the number of patients will more than double the above-mentioned number in the next 20 years. Although there are several therapeutic drugs, it is still desired to develop a treatment method that can be applied to the progressive stage of the disease, or a treatment method that can effectively block its progression at an early stage, and a meth...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09A61K35/76A61K39/00A61K48/00A61P25/28
CPCC07K14/4711A61K2039/53A61K48/00A61K2039/545C12N15/09C12N2760/18843C07K2319/55C12N2799/04C12N2799/021A61K39/0007A61K38/00A61P25/28
Inventor 井上诚佐伯晃一游军田畑寿晃岩崎仁朱亚峰长谷川护
Owner DNAVEC CORP
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