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Compound containing type I collagen and hyaluronic acid as well as preparation and application of compound

A hyaluronic acid and complex technology, applied in the field of biomedicine, can solve the problems of limiting the research and application of collagen-HA biomimetic composite repair materials, affecting the functional activity of collagen, and collagen instability, so as to promote repair and restore blood circulation , easy large-scale production, simple preparation method

Pending Publication Date: 2020-07-17
广州市红十字会医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] HA exists in the form of its acid in the body, and the existing extraction methods are usually obtained in the form of its salt due to technological limitations; collagen and HA in the form of salt have opposite charges, and it is very easy to form a polymer in a liquid state Therefore, this greatly limits the research and application of collagen-HA biomimetic composite repair materials.
In order to expand the use of collagen-HA biomimetic composite repair materials, the methods reported in the existing patent literature include designing the two to be used in isolation (201310099817.4); there are also physical or chemical methods to dissolve and mix the two, such as electrochemical treatment Remove the sodium salt in HA (200910041033.X), but the process is more complicated; thiol-based HA is modified to self-crosslink with collagen to form a three-dimensional hydrogel scaffold (201711462799.6), which will affect the functional activity of collagen
[0006] In addition, the isoelectric point of collagen is about pH 7.4. When the solution is close to neutral, collagen is unstable and easy to precipitate

Method used

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  • Compound containing type I collagen and hyaluronic acid as well as preparation and application of compound
  • Compound containing type I collagen and hyaluronic acid as well as preparation and application of compound
  • Compound containing type I collagen and hyaluronic acid as well as preparation and application of compound

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1: Preparation of collagen and detection of physical and chemical properties

[0064] Pre-treatment: chopped beef tendon, cleaned, washed with normal saline, encapsulated, and treated with Co 60 disinfect. The remaining steps are performed under sterile conditions.

[0065] Preparation of high-purity type Ⅰ collagen solution: add 0.5M acetic acid to the beef tendon, add pepsin (20g beef tendon, add 1g pepsin), enzymolyze at 6°C for 24-48 hours, stop with 15mmol / LEDTA enzymolysis. The supernatant was collected by centrifugation to obtain a type I collagen solution, and this process was carried out at 4°C.

[0066] Purification: Add collagen solution and 10% NaCl for salting out at a ratio of 1:1 by volume, and let stand overnight. The precipitate was obtained by centrifugation, dissolved in 0.5M acetic acid, and dialyzed in deionized water to pH 5 to obtain a high-purity type I collagen solution (≧10mg / ml).

[0067] Standard control product: Sigma type I col...

Embodiment 2

[0071] Embodiment 2: the preparation (pH 6) of this application compound

[0072] a. Preparation of balance substance (90ml): add NaH to deionized water 2 PO 4 0.643g, Na 2 HPO 4 0.189g; NaCl 0.47g, glucose 0.1g, 1,3 propylene glycol 0.5ml, Tween 80 0.1ml.

[0073] b. Measure 10ml of 10mg / ml type I collagen solution.

[0074]c. 800mg of HA is dissolved in 40ml of the balance substance prepared in a, and the concentration of HA is 20mg / ml.

[0075] d. Mix steps a, b, and c to obtain 100ml of a complex containing type I collagen and HA, the pH of the complex is 6, NaH 2 PO 4 The concentration is about 0.643% by weight, Na 2 HPO 4 The concentration of NaCl is about 0.189% by weight, the concentration of NaCl is about 0.47% by weight, the concentration of glucose is about 0.1% by weight, the concentration of 1,3 propanediol is 0.5ml / ml, the concentration of Tween 80 is 0.1ml / ml, I The concentration of type collagen was 1 mg / ml, and the concentration of HA was 8 mg / ml. ...

Embodiment 3

[0076] Example 3: Preparation of the complex of the present application (pH 7)

[0077] a. Prepare balance (70ml) substance: add NaH to deionized water 2 PO 4 0.322g, Na 2 HPO 4 0.566g, NaCl 0.45g, glucose 2g, 1,3 propylene glycol 1.5ml, Tween 80 0.5ml.

[0078] b. Measure 30ml of 10mg / ml type Ⅰ collagen solution.

[0079] c. Dissolve 200mg of HA in 20ml of the balance substance prepared in a, and the concentration of HA is 10mg / ml.

[0080] d. Mix steps a, b, and c to obtain 100ml of a complex containing type I collagen and hyaluronic acid. The pH of the complex is 7, NaH 2 PO 4 The concentration is about 0.322% by weight; Na 2 HPO 4 The concentration of NaCl is about 0.566% by weight, the concentration of NaCl is about 0.44% by weight, the concentration of glucose is about 5% by weight, the concentration of 1,3 propanediol is 1.5ml / ml, the concentration of Tween 80 is 0.5ml / ml, transparent The concentration of hyaluronic acid is 2 mg / ml, and the concentration of t...

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Abstract

The invention provides a compound containing type I collagen and hyaluronic acid. The compound is characterized in that the compound comprises the type I collagen, the hyaluronic acid and a balance substance, wherein the balance substance comprises a NaH2PO4-Na2HPO4 system, a Tris-HCl system, a Tris-maleic acid system or an HEPES system, and comprises NaCl and / or KCl; and the pH value of the compound is 6-8, and the concentration of the NaCl and / or KCl is 0.2-0.9 wt%. The invention also provides a preparation method of the compound. According to the compound disclosed by the invention, in a physiological pH value range, the type I collagen and the hyaluronic acid can be mutually dissolved in a relatively wide proportion, and the system is stable and does not precipitate or denature for a long time; and the osmotic pressure and tension of the compound are suitable for a physiological system. The compound provided by the invention can effectively promote epidermal cells to secrete collagenous fibers, inhibit inflammation and effectively promote skin tissue repair. The preparation method of the compound containing the type I collagen and the hyaluronic acid provided by the invention is simple, low in cost and easy for large-scale production.

Description

technical field [0001] The invention belongs to the field of biomedicine, and specifically relates to a compound containing type I collagen and hyaluronic acid, a preparation method of the compound, and an application of the compound in preparing medicine for repairing skin damage, or the compound Cosmetics used to repair skin damage. Background technique [0002] Skin damage is the defect and loss of skin tissue structure and function caused by internal and external factors. Skin damage can be divided into minor damage and severe damage. Under normal circumstances, minor injuries can heal on their own, but severe injuries cannot achieve good results by relying on self-healing. [0003] The metabolic balance of cells is the basis of skin tissue remodeling and repairing. Exogenous addition of active monomers can improve the microenvironment of damaged skin tissue, promote the synthesis and metabolism of nutrients required for skin tissue metabolism, and accelerate the proce...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/01A61K31/728A61K36/537A61P17/02A61P29/00A61P37/08A61P17/18A61P17/10A61K8/73A61K8/65A61K8/9789A61Q19/00A61Q19/08
CPCA61K38/014A61K31/728A61K36/537A61P17/02A61P29/00A61P37/08A61P17/18A61P17/10A61K8/735A61K8/65A61K8/9789A61Q19/00A61Q19/005A61Q19/08A61K2300/00
Inventor 杨小红朱伟聪刘少杰崔树良
Owner 广州市红十字会医院