Use of cinnamon bark extract for treating amyloid-associated diseases
A technology of amyloid, extract, applied in the application field of Alzheimer's disease
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[0102] A. Preparation of Extract
[0103] The extract was isolated using the following three steps:
[0104] a) Buy cinnamon bark in the market, grind it into powder, and then make it stir overnight in 0.01M-0.02M aqueous phosphate buffer (pH 7.0). The supernatant was separated by centrifugation and used as the crude neutralization extract;
[0105] b) Use 0.15M~0.3M KCl or 0.08M~0.12M MgCl 2 to precipitate the active substance in the crude extract, and dissolve the precipitate in water or 0.01M-0.02M phosphate buffer (pH 7.0);
[0106] c) The extract precipitated solution is optionally loaded into a Sepharose 4B column and eluted as detailed below.
[0107] B. Elution of active fraction
[0108] Use 0.08M~0.12M MgCl 2 Or 0.15M ~ 0.3M KCl to precipitate 60ml of crude extract. The precipitate was dissolved in water or 0.01M-0.02M phosphate buffer, and then loaded onto a 10 ml column of Sepharose 4B washed in advance with 0.01M phosphate buffer (pH 7.0). After loading, th...
Embodiment 1
[0145] Example 1: Determination of the molecular weight of the extract by neutralizing avian influenza H9N2 with the extract obtained as disclosed herein
[0146] A 10 g sample of cinnamon powder (Cinnamon Viet Nam 1696) was extracted in 200 ml of 0.02M, pH 7 phosphate buffer (PB) overnight with stirring. The slurry was centrifuged and the supernatant (168ml in each sample) was dialyzed against water for 4 days in five different bags with different cut-offs from 1KD to 50KD. The material dialyzed out of the bag is concentrated to half of the original volume of the sample in the bag by airflow. Aliquots for hemolysis assays were taken from the fluid inside and outside each bag (twice outside the bag). The results are summarized in Table 1.
[0147] Table 1: Determination of Molecular Weight of Extracts
[0148]
[0149] As shown by the results, approximately 25%-30% of the antiviral activity was lost after dialysis in bags with a cut-off of 25KD. After dialysis in bags w...
Embodiment 2
[0150] Embodiment 2: adopt multiple amount of extracts to inhibit fibril aggregation (EM observation)
[0151] Beta-amyloid (1-40) was mixed with various amounts of extract or with purified extract as described herein above. After 9 days, each mixture was observed using TEM as described.
[0152] Figure 2A β-amyloid (Aβ) derived peptides (Aβ 1-40 ) accumulate into fibrils. The extract concentration was 1 μg / ml ( Figure 2B ) and 10μg / ml ( Figure 2C ) Partial inhibition of fibril formation has been observed. Larger amounts of extract were sufficient to achieve complete inhibition of the fibril formation of the tested β-amyloid ( Figure 2D and 2E ).
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