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New application and preparation method of hyaluronic acid fragment

A technology of hyaluronic acid and a manufacturing method, which is applied in the field of biomedicine, can solve the problems of a stable manufacturing method of bioactive hyaluronic acid fragment B-HA, limited research on new clinical applications of manufacturing principles, etc., and achieves good tissue permeability and safety. The effect of good permeability and good tissue permeability

Pending Publication Date: 2022-08-05
QINGDAO AGRI UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The above-mentioned existing technologies have limited research on the stable manufacturing method, manufacturing principle, mechanism of biological activity, new biological activity and potential new clinical application of the bioactive hyaluronic acid fragment B-HA, and further research and development are needed

Method used

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  • New application and preparation method of hyaluronic acid fragment
  • New application and preparation method of hyaluronic acid fragment
  • New application and preparation method of hyaluronic acid fragment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] OBJECTIVE: To study the method of producing different low molecular weight hyaluronic acid fragments by fully enzymatically hydrolyzing high-molecular hyaluronic acid raw materials with different hyaluronidase enzymes or slightly excess.

[0043] method:

[0044] 1. Production of recombinant human hyaluronidase PH20

[0045] Based on the method described in reference [49], the cDNA of artificially synthesized recombinant human hyaluronidase PH20 was inserted into the GC-rich pMH3 vector to construct the pMH3-PH20 expression vector; then the pMH3-PH20 expression vector was transferred into CHO -S cell line, screened the CHO-S cell line with high expression of PH20, and carried out amplification and large-scale culture in the rapid flow animal cell reactor; the harvest liquid containing PH20 was filtered through a 0.22 μm filter membrane, and then passed through a QFF chromatography column, Phenyl HP hydrophobic chromatography column, CHT I type ceramic hydroxyapatite ch...

Embodiment 2

[0089] Objective: To study the rapid absorption pathway of hyaluronic acid fragment (hereinafter referred to as B-HA or HA35) with an average molecular weight of 35kDa (hereinafter referred to as B-HA or HA35) after subcutaneous and intravenous injection in mice after labeling with 125-I and 99mTc, and further use in vivo molecular imaging technology and human fresh extraction of human neutrophils and mononuclear cells (mainly lymphocytes and a small number of monocytes) to study their new therapeutic effects and new mechanisms of action.

[0090] Laboratory reagents, equipment, human and animal blood cell samples and laboratory animals:

[0091] 1. Experimental reagents: injection-grade hyaluronic acid raw materials with an average molecular weight of 1600kDa, common hyaluronic acid raw materials with an average molecular weight of 300kDa and hyaluronic acid fragments with an average molecular weight of 24kDa were purchased from Bloomage Freda Biomedical Co., Ltd., and the ave...

Embodiment 3

[0142] Objective: To use commercial bovine hyaluronidase PH20 injection and macromolecular hyaluronic acid injection to produce B-HA or HA35 with an average molecular weight of 35.4kDa in a short time to treat hypersensitivity of local large-area redness, swelling and hard pain caused by mosquito bites. Inflammation.

[0143] method:

[0144] Twelve cases of patients with hypersensitivity reaction to mosquito (mosquito or bumblebee) bites (with rapid localized large-area redness, swelling and pain in the bite area). Use the following 37 degrees for 20 minutes to mix the two injections and inject into the affected area outside the instructions, and then observe the change of the large area of ​​red, swollen and hard pain in the bite area after treatment.

[0145] Before injection, bovine testicular hyaluronidase PH20 (1500u / piece, H31022111, Shanghai No. 1 Biochemical Pharmaceutical Factory) and sodium hyaluronate (hyaluronic acid) injection (trade name Speite) with an average...

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Abstract

The invention discloses an application of a hyaluronic acid fragment as a human mononuclear cell migration accelerant, an application of the hyaluronic acid fragment as an immunomodulator of human lymphocytes and liquid reflux and lymphocytes through subcutaneous or intravenous injection, and an application of the hyaluronic acid fragment as a human neutrophil migration inhibitor. The hyaluronic acid fragment is prepared by sufficiently performing enzymolysis on a hyaluronic acid raw material with the pH value of 20, can pass through a 0.22 mu m filter membrane and has the average molecular weight of 35 + / -8KDa. The invention also discloses application of the hyaluronic acid fragment in preparation of drugs for treating inflammatory diseases related to human mononuclear cell and neutrophil migration, and application of the hyaluronic acid fragment in preparation of drugs for treating inflammatory diseases related to human lymphocyte and liquid backflow. The invention also discloses a preparation method of the hyaluronic acid fragment and a method for preparing hyaluronic acid fragments with different molecular weights. The invention finds a new preparation method, action mechanism and application of the hyaluronic acid fragment, and lays a foundation for the pharmaceutical research of the hyaluronic acid fragment.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a new application and manufacturing method of a hyaluronic acid fragment. Background technique [0002] At present, the scientific society still does not have a unified view on the physiological function and pharmaceutical efficacy of the human extracellular matrix hyaluronic acid. The earliest hyaluronic acid products were injections for the treatment of arthritis with an average molecular weight greater than 1200kDa, such as Durolane and Synvisc. Literature studies have shown that the physiological and pharmacological effects of hyaluronic acid are related to its molecular weight. Literature studies have also shown that the hyaluronic acid fragment with an average molecular weight of 35kDa extracted from human colostrum has various physiological functions and pharmacological effects. The hyaluronic acid fragment with an average molecular weight of 35 kDa extracted from human colost...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/728A61P29/00C12P19/14C12P19/04
CPCA61K31/728A61P29/00C12P19/14C12P19/04A61K9/0019
Inventor 郭田田王家麒李鑫荣贾潇潇吴书音惠鑫瑶桐辉崔家友张之辉丛振昱王起飞王凤舞宋琳惠觅宙
Owner QINGDAO AGRI UNIV
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