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Injection liquid containing small molecular hyaluronic acid and application thereof

A technology of hyaluronic acid and human hyaluronidase, applied in the field of disease treatment drugs, can solve the problems of complex, difficult to determine clinical indications, difficult to predict the success of clinical research and druggability of small molecule hyaluronic acid, etc. , to achieve the effect of convenient production and stable preparation

Inactive Publication Date: 2017-10-31
惠觅宙
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the recent patent WO / 2014 / 165713 of CEDARS-SINAI MEDICAL CENTER shows that the small molecule hyaluronic acid produced by bacterial streptococci hyaluronidase cutting less than 10KD does not cause inflammation, but the bacterial Streptomyces hyaluronidase produces more than 10KD However, the small molecule hyaluronic acid caused inflammation, suggesting that the function of small molecule hyaluronic acid may also be related to the type of cutting enzymes
[0022] The above contradictory research results suggest that the exact function of the small-molecule hyaluronic acid fragment is currently difficult to determine, and the production method, molecular weight, type of cutting enzyme, and use of the small-molecule hyaluronic acid fragment may affect its functional performance. In addition to these known possible factors, there are still many other unknown factors that researchers are not sure about.
Therefore, although people in the industry pin their hopes on the medical value of small molecule hyaluronic acid, it is still difficult to predict the success of clinical research and druggability of small molecule hyaluronic acid, especially to determine the clinical indications
Although our previous research obtained a feasible and effective small-molecule hyaluronic acid in vitro preparation, due to the complex internal environment of the human body, the in vivo injection preparation of small-molecule hyaluronic acid may be completely different from the in vitro preparation. The effects in humans and animals may not be the same, therefore, it needs to be verified by clinical research

Method used

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  • Injection liquid containing small molecular hyaluronic acid and application thereof
  • Injection liquid containing small molecular hyaluronic acid and application thereof
  • Injection liquid containing small molecular hyaluronic acid and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Objective: To study the production process of injections, and to manufacture injections with and without recombinant human hyaluronidase PH20 residual protein.

[0076] Method: Using animal cells (CHO cells) to produce glycosylated recombinant human hyaluronidase PH20, including: artificially synthesizing the gene cDNA of glycosylated recombinant human hyaluronidase PH20, inserting GC-rich pMH3, pMH4, pMH5 Empty expression vector, build up pMH3-PH20, pMH4-PH20, pMH5-PH20 expression vector (adopting the technology construction expression vector described in the patent document of CN102124019A here to express recombinant protein so that pMH3-PH20, pMH4- The cDNA expression vectors of PH20 and pMH5-PH20 were transferred into CHO-S cell line, and the CHO-S cell line with high expression of PH20 was screened. figure 1 The scale-up and large-scale cultivation of the torrent-type animal cell reactor is shown; the harvest liquid containing PH20 is filtered at 0.22um, and then s...

Embodiment 2

[0085] Objective: To study the anti-inflammatory and antibacterial effects of Hyaluronic Acid Injection 1 of Example 1 in vitro experiments, molecular cytology experiments, and small animal experiments.

[0086]Method: The Hyaluronic Acid Injection 1 of Example 1 diluted 1-fold with normal saline was used to conduct in vitro antibacterial experiments, molecular cytology experiments, and small animal experiments to detect the anti-inflammatory and antibacterial effects of Hyaluronic Acid Injection.

[0087] result: Figure 4 It shows that hyaluronic acid injection inhibits the growth of Porphyromonas gingivalis colony in vitro, suggesting that hyaluronic acid injection materials are not conducive to the growth of Porphyromonas gingivalis (Note: the negative control is normal saline, and the positive control is antibiotics Metronidazole); Figure 5 Show the anti-inflammatory and antibacterial effects of hyaluronic acid injection on skin wounds of small animals; Image 6 It is ...

Embodiment 3

[0090] Purpose: To study the beauty and anti-aging effects of two kinds of hyaluronic acid injections containing (experimental group 1) and not (experimental group 2) hyaluronic acid fragments of recombinant human hyaluronidase PH20 residual protein in the above embodiment 1 efficacy and safety.

[0091] Method: Use the injection solution containing hyaluronic acid fragments of recombinant human hyaluronidase PH20 residual protein (100mg of hyaluronic acid fragments with a molecular weight of 10KD-60KD, 115-125mM sodium chloride, 1mM magnesium ions, no degraded hyaluronic acid The active recombinant human hyaluronidase residual protein is less than 20 micrograms) is the experimental group 1, which is used for various diseases or subclinical problems frequently occurring or aging and aging-related diseases frequently occurring in China's current air pollution, drinking water pollution and work pressure environment 38 subjects (average age 61±26 years old, 20 males, 18 females) ...

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Abstract

The invention discloses an injection liquid containing small molecular hyaluronic acid and an application thereof. The injection liquid is prepared by the following steps: A, adopting animal cell or yeast or plant expression produced diastatic recombinant human hyaluronidase PH20; B, preparing a macromolecular hyaluronic acid solution by using a macromolecular hyaluronic acid raw material, introducing sodium chloride and magnesium ions, and adding the diastatic recombinant human hyaluronidase PH20 to cut to obtain a reaction liquid containing a hyaluronic acid fragment, the molecular weight of which is 10-60KD; C, deactivating the residual diastatic recombinant human hyaluronidase PH20; and D, performing bacterial filtration, virus filtration and deactivation and ultrafiltration and concentration. The injection liquid disclosed by the invention not only has the effects of beautifying face and being aging-resistant to the face, but also accidentally shows treatment effects of quickly reliving pain and itching, increasing the physical power and energy, reducing subcutaneous fat of head, face and neck and treating various diseases and disease states.

Description

technical field [0001] The invention relates to the field of medicines for treating diseases, in particular to an injection containing small molecule hyaluronic acid and its use for treating various diseases. Background technique [0002] Hyaluronic acid, also known as hyaluronic acid, uronic acid, hyaluronic acid, English name hyaluronic acid (referred to as HA), is a high-level polysaccharide composed of D-glucuronic acid and N-acetylglucosamine, and the number of disaccharide units can reach 25,000 you. Tissues such as human subcutaneous tissue, epidermis and oropharyngeal mucosa contain a large amount of macromolecular hyaluronic acid. In other words, macromolecular hyaluronic acid is the basic building material of various tissues such as human skin, mucous membranes, and subcutaneous tissue, and has the functions of water retention and moisturizing. The existing hyaluronic acid products in the market are mainly facial care or facial beauty products with the addition o...

Claims

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Application Information

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IPC IPC(8): A61K9/08A61K31/728A61P27/02A61P29/00A61P17/04A61P19/04A61P21/00A61P19/08A61P15/00A61P35/04A61P35/00A61P9/10A61P17/00A61P39/06A61P3/04A61P1/02A61P11/04A61P9/00
CPCA61K9/0019A61K9/08A61K31/728C12P19/26Y02P20/59
Inventor 惠觅宙
Owner 惠觅宙
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