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A compound vaccine for preventing and treating senile dementia and its preparation method

A technology for senile dementia and vaccines, applied in drug combinations, gene therapy, pharmaceutical formulations, etc., can solve problems such as low antibody titers and side effects, achieve simple production methods, low cost, and suppress meningitis side effects Effect

Active Publication Date: 2014-10-29
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For this reason, researchers have proposed different solutions in continuous research and discussion, for example, adding non-steroidal anti-inflammatory drugs, antiviral drugs or hormones to prevent inflammation during vaccine treatment. In addition, There is also the application of Aβ subunit vaccines to remove the toxic fragments of Aβ, which can suppress adverse cellular reactions while producing antibodies; but the above methods have their disadvantages, such as hormones and other inhibitors also have side effects, subunit vaccines, The antibody titers produced are not very high, etc.

Method used

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  • A compound vaccine for preventing and treating senile dementia and its preparation method
  • A compound vaccine for preventing and treating senile dementia and its preparation method
  • A compound vaccine for preventing and treating senile dementia and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1 Preparation of Alzheimer's Disease Aβ42 Nucleic Acid Vaccine

[0067] 1) Construction of eukaryotic expression plasmid for Alzheimer's disease Aβ42

[0068] Using the plasmid Abeta 42-C3d3 (provided by Dr. McArgo Janie) as a template, primer P1: 5'-AAAGGATCCATGGATGCAGAATTCC-3' and primer P2: 5'-GCCTCTAGATTACGCTATGACAACA-3', (in primer 1 and primer 2, respectively The Aβ42 gene was amplified by PCR under the guidance of BamHI recognition site and XbaI recognition site). Reaction system: 1 μL plasmid template, 10 pmol each of primer 1 and primer 2, 500 mM KCl, 100 mM Tris-HCl (pH8.4), 1.5 mM MgCl 2 , 100μg / mL BSA, 1mM dNTPs, 2.5U Taq DNA polymerase, the total volume is 25μL; the reaction conditions are: denaturation at 94°C for 30 seconds, renaturation at 60°C for 30 seconds, extension at 72°C for 30 seconds, a total of 30 cycles; The amplified DNA fragments in 1.5% liposugar gel electrophoresis were recovered and connected to the pMD18-T cloning vector; the c...

Embodiment 2A

[0071] Example 2 Prokaryotic expression of Aβ42 protein

[0072] 1) Construction of prokaryotic expression plasmid for Alzheimer's disease Aβ42

[0073] Using Abeta 42-C3d3 as a template, in primer 1: 5'-AAAGGATCCATGGATGCAGAATTCC-3' and primer 2: 5'-GCCGTCGACTAACGCTATGACAACA-3' (in primer 1 and primer 2, BamHI recognition site and SalI recognition site were introduced respectively) The Aβ42 gene was amplified by PCR under the guidance of the guide; the recovered target fragment was connected to the pMD18-T vector, and after the correct identification by enzyme digestion, the target fragment was subcloned into the pET28a vector. same. The result is as image 3 As shown, A is the result of double digestion of pMD18T-Aβ42 plasmid with BamHI and SalI, B is the result of double digestion of pET28a-Aβ42 plasmid with BamHI and SalI, and the sequence analysis result is correct.

[0074] Using pMD18T-Aβ42 as a template, in primer 1: 5'-AAAGGATCCATGGATGCAGAATTCC-3' and primer 2: 5'-G...

Embodiment 3A

[0077] Example 3 ELISA detection and T cell response detection of antibody production after Aβ42 protein vaccine and DNA vaccine co-immunized with different mouse species

[0078] Select 6-8 weeks old mice of Balb / c and C57BL / 6 species for immunization, and detect whether the combined immunization of Aβ42 protein vaccine and DNA vaccine can cause immunosuppression by detecting antibody IgG and T cell proliferation responses, and At the same time, it does not affect the production of antibodies.

[0079] 1) ELISA detection of antibodies produced after Aβ42 protein vaccine and DNA vaccine were co-immunized with Balb / c and C57BL / 6 mice

[0080] Divide 16 6-8 week-old BALB / c or C57BL / 6 female mice into 4 groups, 4 in each group; the first group was intramuscularly injected with 50 microliters of PBS solution containing 50 micrograms of pVAX1-Aβ42 plasmid DNA; Group subcutaneous immunization contained 50 micrograms of one copy of Aβ42 protein, 1 / 2 volume of Freund's complete adjuv...

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Abstract

Disclosed herein are vaccines comprising an amyloid beta antigen and a nucleic acid encoding the amyloid beta antigen, or a variant thereof. T cell proliferation can be inhibited and iTreg cells can be stimulated in the subject administered the vaccine. Further provided are methods for treating Alzheimer's Disease, methods for reducing or slowing the rate of formation of amyloid beta plaques, methods for treating encephalitis, and methods for of reducing brain swelling. The methods can comprise administering the vaccine to a subject in need thereof.

Description

technical field [0001] The invention belongs to the field of biological products, and relates to a compound vaccine for preventing and treating senile dementia and a preparation method thereof. Background technique [0002] Dementia is a brain disorder that is caused by abnormal degeneration of the brain. According to statistics, most of the patients are elderly. The clinical manifestations are: due to the decline of brain function, the patients are affected in terms of memory, calculation, learning, understanding, and even language, judgment, and sense of direction. The disease not only suffers from the patient himself, but also has a very negative impact on his family and even the whole society. With the increase of the elderly, the number of patients with Alzheimer's disease is also increasing. Alzheimer's disease (AD) is a common form of degenerative dementia. At present, there are more than 25 million AD patients in the world. Due to the characteristics of its onset...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/00A61K48/00A61P25/28A61P37/02
CPCA61K39/0005A61K31/7088C07K14/4711A61K38/00C07K16/18A61K45/06A61P25/28A61P37/02
Inventor 王宾于杨朱贤主王爽
Owner FUDAN UNIV
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