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Application of tea polypeptides in the preparation of drugs to improve, alleviate or treat gout

A technology of polytea and medicine, which is applied in the pharmaceutical use of tea polypeptide, and the application field of tea polypeptide in the preparation, improvement, alleviation or treatment of gout drugs, which can solve the problems of easy drug resistance and other problems

Pending Publication Date: 2019-06-21
WUYI UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, compared with the existing therapeutic drugs that need to be taken for a long time, treat the symptoms but not the root cause, and are prone to drug resistance and many other problems, tea polypeptide can not only treat general gout caused by living habits, but also can target the treatment of gout caused by ABCG2Q141K. Gout disease caused by genetic mutation, wider range of treatment and better treatment effect

Method used

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  • Application of tea polypeptides in the preparation of drugs to improve, alleviate or treat gout
  • Application of tea polypeptides in the preparation of drugs to improve, alleviate or treat gout
  • Application of tea polypeptides in the preparation of drugs to improve, alleviate or treat gout

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Embodiment 1, the preparation of tea polypeptide sample

[0059] 1. Preparation of tea protein:

[0060] S1. Wall breaking: use enzymatic hydrolysis technology to enzymatically hydrolyze and break the green tea tea dregs;

[0061] S2. Alkaline extraction: continue to extract green tea protein from the green tea dregs after enzymatic hydrolysis in step S1 by an alkaline method to obtain a green tea protein extract;

[0062] S3. Acid precipitation: acid precipitation is carried out on the green tea protein extract obtained in S2 to obtain a precipitate;

[0063] S4. Desalting: desalting the precipitate obtained in S3;

[0064] S5. Decolorization: take the desalted green tea protein in S4, use 75% acetone to decolorize, and freeze-dry to obtain green tea protein;

[0065] S6. Enzymolysis: the green tea protein obtained in S5 is added to food protease for enzymolysis to obtain a green tea protein polypeptide with blood pressure lowering activity.

[0066] Wherein, the g...

Embodiment 2

[0078] Example 2. Preparation and cell transfection of ABCG2 Q141K

[0079] 1 site-directed mutagenesis

[0080] 1.1 Primer design

[0081] The design principles to be followed for site-directed mutagenesis are: change as few sites as possible, reduce mutation frequency, and be easy to verify. At the mutation site, a pair of reverse complementary primers was designed.

[0082] 1) Primer synthesis and desalting must be complete, and it is best to choose a purer purification scheme; 2) Primers generally need to be about 25-35bp, and the end of the primer ends with G or C; 3) Refer to the codon comparison table to minimize base changes; 4 ) There are at least 5-9 sites matching the template at both ends of the mutated site; 5) When designing primers, the restriction site on the mutation primer sequence can be considered, and the restriction site is preferably a vector and a gene fragment Restriction sites not found elsewhere. At the same time, the restriction site can be used...

Embodiment 3

[0119] Embodiment three, Western blot detection protein expression level

[0120] 1 Extraction of total cell protein

[0121] (1) Pretreatment of cells: by 37°C CO 2 Take out the 293T cells cultured in the cell culture plate in the incubator, discard the original culture medium, and wash the cell surface 3-4 times with 4°C pre-cooled 1×PBS to remove the serum and dead cells in the medium, and try as much as possible. Aspirate the remaining PBS in the cell culture dish. The washed cells were directly frozen and stored in a -80°C refrigerator for protein extraction. This method preserves cells for at least two weeks. (Note: If necessary, after washing the cells with PBS, it is not necessary to freeze them at -80°C, and you can directly proceed to step 2)

[0122] (2) Add PMSF-containing lysate: Take out the cell culture dish from the -80°C refrigerator, place it on ice, add PMSF-containing cell lysate to each of the above-mentioned culture dishes, and shake gently to make th...

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Abstract

The invention provides application of tea polypeptides in restoring mutation expression of ABCG2 Q141K to improve gout caused by AGCG2 Q141K mutation. The technique herein belongs to the field of teahealth and functionality; the invention particularly relates to researches on the health functions of tea active ingredients.

Description

technical field [0001] The invention relates to the field of medicines, in particular to the pharmaceutical use of tea polypeptides, especially the application of tea polypeptides in the preparation of medicines for improving, alleviating or treating gout. Background technique [0002] Gout is a long-term disorder of purine metabolism, and the blood uric acid level is maintained at a high level for a long time, which leads to a series of clinical syndromes of tissue damage or mutation. Its real-life manifestation is that after uric acid forms a crystal structure, it gradually accumulates in tissues, organs and joints, leading to gout deposition, intermittent attacks of acute or chronic arthritis, and even joint distortion, often accompanied by kidney damage, such as Chronic tubulointerstitial nephritis and uric acid kidney stones and other symptoms. The pathogenesis of gout is due to the excessive production of uric acid and the obstruction of uric acid excretion in the kid...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K1/14A61K38/01A61P19/06
Inventor 李冬利安然张文姬孙世利黎秋华赖幸菲孙伶俐张焜
Owner WUYI UNIV
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