Anti-aβ31-35 antibody for treating and preventing Alzheimer's disease and preparation method thereof

An Alzheimer's disease and antibody technology, applied in the field of anti-Aβ31-35 antibody and its preparation for the treatment and prevention of Alzheimer's disease, achieving the effect of small side effects and strong specificity

Inactive Publication Date: 2011-11-30
SHANXI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Therefore, clinical trials are currently suspended

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  • Anti-aβ31-35 antibody for treating and preventing Alzheimer's disease and preparation method thereof
  • Anti-aβ31-35 antibody for treating and preventing Alzheimer's disease and preparation method thereof
  • Anti-aβ31-35 antibody for treating and preventing Alzheimer's disease and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Preparation steps of anti-Aβ31-35 antibody for treating and preventing Alzheimer's disease:

[0016] (1) Antigen preparation

[0017] The Aβ31-35 fragment was synthesized by organic solid-phase synthesis method, the solid phase carrier was HMP resin, the amino acid protected by Fmoc (9-fluorenylmethyloxycarbony) was used to synthesize the amino acid from the carboxyl end of the polypeptide to the amino end, and the peptide resin was cleaved by TFA method. Crude Aβ31-35 fragments were obtained after pressure distillation and ether extraction. It was then purified by high-performance liquid chromatography (HPLC), and the purified sample was analyzed by HPLC. The purity was 95%, and the purified Aβ31-35 fragment was freeze-dried.

[0018] Take the purified Aβ31-35 fragment and couple with hemocyanin. The coupling method is to use the carbodiimide method [1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide, EDC] for condensation reaction to obtain Aβ31-35 Fragment-hemocyanin c...

Embodiment 2

[0024] We observed the protective effect of anti-Aβ31-35 antibody on brain function or cell damage caused by intracerebroventricular injection of Aβ or addition of Aβ to neuronal cell culture. The main results are as follows:

[0025] (1) Morris water maze experiment figure 2 The average latency (A) and typical swimming trajectories (B) of rats in each group to find the underwater platform are shown. Compared with normal rats, intracerebroventricular injection of 5 nmol Aβ1-42 can significantly prolong the platform-seeking latency; after treatment with different concentrations of anti-Aβ31-35 antibody, the latency and swimming distance are shortened, especially 0.5 and 5 nmol anti-Aβ31-35 antibody pre-treatment. The treatment group was significantly smaller than the Aβ1-42 group on each training day (P<0.01), indicating that the behavioral damage caused by Aβ can be reversed by the anti-Aβ31-35 antibody.

[0026] (2) In vivo hippocampal LTP pre-experiment such as image 3 ...

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Abstract

The invention provides an anti-Aβ31-35 antibody for treating and preventing Alzheimer's disease (AD). The preparation method: synthesize Aβ31 whose amino acid sequence is Ile-Ile-Gly-Leu-Met by an organic solid-phase synthesis method The -35 fragment was purified, analyzed, and conjugated with hemocyanin, and the conjugated product was used to obtain anti-Aβ31-35 antibodies after basic immunization and booster immunization of white rabbits. The antibody is used in in vivo AD animal experiments or clinical treatment of AD patients, which can effectively avoid the binding site of the transmembrane part of APP and selectively remove extracellular Aβ. Compared with traditional antibodies against Aβ N-terminus, anti-Aβ31-35 antibodies will have the characteristics of less side effects and stronger specificity in the prevention and treatment of AD.

Description

technical field [0001] The invention relates to an anti-Aβ antibody, in particular to an anti-Aβ31-35 antibody for treating and preventing Alzheimer's disease and a preparation method thereof. Background technique [0002] Alzheimer's disease (AD), also known as senile dementia, is a primary degenerative disease of the central nervous system, mainly manifested as progressive cognitive dysfunction, loss of learning and memory, and personality changes. Severe dementia develops later. The most typical pathological feature of AD is the appearance of a large number of high-density senile plaques in the brain, and its main component is amyloid β protein (Aβ). The neurotoxic effects of Aβ have been widely reported. Such as figure 1 As shown, Aβ comes from transmembrane amyloid precursor protein (Amyloid precursor protein, APP), but APP itself has a neurotrophic effect. Traditional AD immunotherapy mostly uses antibodies against the N-terminus of Aβ. Although it has a certain cu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18A61K39/395A61P25/28
Inventor 祁金顺蔡红艳杨威王昭君王晓晖武美娜
Owner SHANXI MEDICAL UNIV
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