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Novel application of hyaluronic acid fragment and stable making method

A technology of hyaluronic acid and hyaluronidase, applied in the field of biomedicine, can solve the problems such as the stable manufacturing method of bioactive hyaluronic acid fragment B-HA, the limited research on the new clinical application of the manufacturing principle, etc.

Inactive Publication Date: 2020-06-09
QINGDAO AGRI UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In summary, the above 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 is needed. Research and Development

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  • Novel application of hyaluronic acid fragment and stable making method
  • Novel application of hyaluronic acid fragment and stable making method
  • Novel application of hyaluronic acid fragment and stable making method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Objective: To study the stable manufacturing method and principle of using different hyaluronidases to fully or slightly excessively enzymatically hydrolyze high or medium molecular hyaluronic acid raw materials to produce low molecular hyaluronic acid fragments.

[0053] method:

[0054]1. The methods for measuring the molecular weight of hyaluronic acid and hyaluronic acid fragments used in the present invention include electrophoretic measurement methods and 18-angle laser measurement methods. Manufacturers of high or medium molecular weight hyaluronic acid raw materials and the present invention adopt viscometer and 18-angle laser to measure.

[0055] 2. Hyaluronic acid raw materials: hyaluronic acid raw materials with an average molecular weight of 300kDa (medium molecular weight) and 1600kDa (high molecular weight) (Huaxi Freda Biotechnology Co., Ltd.).

[0056] 3. Hyaluronidase: recombinant human hyaluronidase PH20 (Shaoxing Huihui Technology Co., Ltd.), bovine ...

Embodiment 2

[0103] Objective: To study the effect of using recombinant human hyaluronidase PH20 in sufficient or slight excess to fully enzymatically hydrolyze high-molecular-weight and medium-molecular-weight hyaluronic acid raw materials to stably produce hyaluronic acid fragments (hereinafter referred to as B-HA) with an average molecular weight of 35±8KDa. Effects of extracted human leukocytes (neutrophils and mononuclear cells) on removal, phagocytosis, apoptosis, TNFɑ secretion, IL-6 secretion and beta-defensin secretion.

[0104] result:

[0105] 1. The effect of B-HA on the secretion of inflammatory factor TNFɑ from neutrophils

[0106] Table 1. B-HA did not significantly promote or inhibit the secretion of TNFɑ from human neutrophils in the presence or absence of erythrocytes (p>0.05; p>0.05). Similar results were obtained using freshly extracted human neutrophils from different healthy volunteers.

[0107] Table 1. Effect of B-HA on secretion of TNF by human neutrophils in the...

Embodiment 3

[0176] Objective: To use commercial 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 period of time to treat the high-sensitivity inflammatory response of local large-area redness, swelling and hard pain caused by mosquito bites .

[0177] method:

[0178] 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 injections to inject into the affected area outside the instructions, and then observe the changes in the large area of ​​red, swollen and hard pain in the bite area after treatment.

[0179] Before injection, the recombinant human hyaluronidase PH20 injection (Hylenex, 150unit / mL injectionsolution, Hylozyme Inc) and Durolane 20mg / ml, 3ml / Vial (Genvrier) with an average molecular weight of 1600kDa were prepared according to the ra...

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Abstract

The invention discloses application of a hyaluronic acid fragment as a human neutrophil migration inhibitor, a human mononuclear cell migration inhibitor, a human mononuclear cell apoptosis promoter and / or a human neutrophil beta sozin secretion promoter. The hyaluronic acid fragment is hyaluronic acid fragment which is of a use structure made through sufficient or slightly excessive sufficient enzymolysis of a high or medium molecular weight hyaluronic acid raw material by using hyaluronidase PH20 with two binding zones, and the hyaluronic acid fragment has an average molecular weight of 35+ / -8KDa. The invention further discloses application of the hyaluronic acid fragment in preparing a medicine for treating neutrophil and mononuclear cell mediated inflammation diseases. The invention further discloses a stable making method of the hyaluronic acid fragment and a combined application method of a hyaluronidase injection and a hyaluronic acid injection made on the basis of the stable making method. Through research, the invention finds the stable making method, making principles and anti-inflammation mechanisms of the hyaluronic acid fragment, and bases are made for medicinal research on the hyaluronic acid fragment.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a new application and stable production method of hyaluronic acid fragments. Background technique [0002] At present, the scientific society still does not have a unified view on the physiological function and pharmaceutical efficacy of 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 research shows that the physiological and pharmaceutical effects of hyaluronic acid are related to its molecular weight (1). Literature research also shows that hyaluronic acid fragments with an average molecular weight of 35kDa extracted from human colostrum have various physiological functions and pharmaceutical effects (2, 3, 4, 5, 6, 7). The hyaluronic acid fragment with an average molecular weight of 35kDa extracted from hum...

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 王凤舞惠鑫瑶贾潇潇郭田田王家麒吴书音薄乐崔家友张之辉丛振昱王起飞李鑫荣宋琳惠觅宙
Owner QINGDAO AGRI UNIV
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