Preparation of a hyaluronic acid matrix that can be sterilized by irradiation

Abstract

The present invention provides a hyaluronic acid matrix that can be sterilized by irradiation, which is composed of the following ingredients: 20-30 parts of sodium hyaluronate, 20-30 parts of glycerol, 8-15 parts of carbomer, 3-8 parts of olive oil, 1-5 parts of dipotassium glycyrrhizinate, and 900+n parts of water; wherein n is 10 times the number of dipotassium glycyrrhizinate. The hyaluronic acid matrix provided by the present invention is adjusted by adjusting the formula, and the preparation method is optimized accordingly, and the irradiation sterilization method is adjusted, and the hyaluronic acid matrix obtained is a gel matrix, and the stability meets the requirements of 2-year quality assurance, and the salt tolerance and stability of the gel matrix containing carbomer are improved, and a good dynamic viscosity can be maintained under certain salt concentration conditions, and it can be better applied to production and clinical.

Description

Technical Field

[0001] The invention belongs to the field of gel matrix materials, and in particular relates to the preparation of a hyaluronic acid matrix that can be sterilized by radiation. Background Art

[0002] Hyaluronic acid is the main component of the extracellular matrix and is mainly distributed in the joint cavity, skin, and blood. Hyaluronic acid has high viscoelasticity, plasticity, strong water absorption and moisturizing properties, and good biocompatibility. Based on the above characteristics, hyaluronic acid is widely used as a basic ingredient in medicines, medical devices, and cosmetics.

[0003] Hyaluronic acid matrix products have certain requirements for sterility assurance level (SAL) during actual production, which can be achieved through moist heat sterilization. For invasive surface application products and in vivo application medical products, the SAL of products sterilized by moist heat shall not be greater than 10 -6 . However, moist heat sterilization will affect the performance of hyaluronic acid-based products due to thermal effects, and since hyaluronic acid is a macromolecular material, filtration sterilization cannot be adopted. Cobalt source and electron beam irradiation are common sterilization methods in medicines, medical devices, and cosmetics due to their rapidity, high efficiency, and no residue. When irradiation is used to sterilize hyaluronic acid-based medical products, the β-1,3 and β-1,4 glycosidic bonds in the hyaluronic acid molecules will be broken, resulting in a decrease in its molecular weight and affecting its performance, and this effect increases with increasing dose.

[0004] Based on the above situation, although the commonly used radiation dose can sterilize hyaluronic acid matrix products to a sterility assurance level, it also affects their application because radiation reduces product performance.

[0005] Chinese patent 201710289385.1 discloses a method for aseptic production of hyaluronic acid products, in which 0.01%-0.5%wt of preservatives are added to the hyaluronic acid products and then irradiated, with an irradiation dose of 6-10kGy. By controlling the production of raw materials, water, environment, and packaging materials, adding an appropriate amount of preservatives to the product, and using a lower dose of cobalt 60 irradiation to achieve the sterility standard, it can prevent the degradation of high molecular weight hyaluronic acid, keep the molecular weight of hyaluronic acid within a reasonable ratio, and improve the product quality level. However, no further research has been conducted on other properties of the product, and the addition of preservatives has a certain impact on the addition of subsequent ingredients, and it also has certain harm to the human body.

[0006] Chinese patent 201811540735.8 discloses an irradiated cross-linked hyaluronic acid hydrogel medical cold compress, which, calculated by mass percentage, includes the following components: 8%-10% palm petiole extract, 5%-8% cross-linking agent, 3%-5% cellulose gum, 0.1%-7% hyaluronic acid, and water sufficient to fully dissolve the other components. In this way, the use time of the hydrogel product can be extended from the original 45 minutes to about 2 hours, which can quickly enhance skin moisture, increase skin elasticity, enhance skin smoothness and reduce skin wrinkles, and can achieve better results in postoperative repair of various cosmetic surgeries and daily skin care. However, it is a specific product that mainly focuses on efficacy research and has little effect on improving the gel matrix. Summary of the invention

[0007] In order to solve the above problems, the present invention provides a hyaluronic acid matrix that can be sterilized by radiation and a preparation method thereof. The method reduces the radiation sterilization dose and the effect of radiation on the molecules of hyaluronic acid by adjusting the component formula of hyaluronic acid and controlling the bioload of the hyaluronic acid matrix, thereby ensuring that its performance meets production and clinical needs.

[0008] The technical solution adopted by the present invention is to achieve the expected sterilization and aseptic assurance level of sterile medical device products by combining heat treatment with radiation sterilization, while at the same time ensuring that the matrix of the device product will not undergo quality changes.

[0009] The microbial level of a medical product is its bioburden. According to "Radiation Sterilization of Healthcare Products - Part 2 - Establishing the Sterilization Dose" (GB18280.2-2015 / ISO11173.2-2006), it can be seen that the bioburden and sterilization dose are directly proportional. When the bioburden is 0.1, the sterility assurance level (SAL) = 10 is achieved. -6 When the irradiation dose is 11.0 kGy, the sterility assurance level (SAL) is 10 when the bioburden is 1.0. -6 When the bioburden is 10, the sterility assurance level (SAL) is 10. -6 When the bioburden is 100, the sterility assurance level (SAL) is 10. -6 When the irradiation dose is 21.2 kGy, the sterility assurance level (SAL) is 10 when the bioburden is 1000. -6 The irradiation dose is 24.9 kGy; that is, by controlling the bioburden level of medical products, the dose of irradiation sterilization can be reduced, the impact of irradiation on the hyaluronic acid matrix can be reduced, and the performance of the product can be ensured to meet clinical needs.

[0010] Studies have shown that the effect of temperature on hyaluronic acid is in direct proportion. When heated at 60°C for 90 minutes, the dynamic viscosity of the hyaluronic acid solution does not change; when heated at 80°C for 90 minutes, the dynamic viscosity of the hyaluronic acid solution does not change; when heated at 100°C for 90 minutes, the dynamic viscosity of the hyaluronic acid solution decreases with time; when heated at 121°C for 3.5 minutes, the dynamic viscosity of the hyaluronic acid solution drops sharply.

[0011] Pasteurization shows that heating milk at 62°C-65°C for 30 minutes can achieve a sterilization efficiency of 97.3-99.9%. Therefore, the present invention reduces the bioburden of the hyaluronic acid matrix through appropriate temperature treatment to achieve a lower irradiation dose to ensure no effect on the matrix performance, which is an effective method to achieve a product sterility assurance level.

[0012] The hyaluronic acid matrix has a molecular weight of 1000kDa-5000kDa and a concentration of 0.5%-10%.

[0013] Irradiation sterilization dose: the irradiation dose is 8kGy-11kGy.

[0014] By measuring the sterility assurance level and dynamic viscosity of the hyaluronic acid matrix after sterilization, it is confirmed whether it meets clinical needs.

[0015] In addition, the present invention also found that adding olive oil and dipotassium glycyrrhizinate to the hyaluronic acid matrix of the present invention can effectively improve stability and extend shelf life.

[0016] In one aspect, the present invention provides a hyaluronic acid matrix that is sterilizable by radiation.

[0017] Specifically, the hyaluronic acid matrix is ​​composed of the following ingredients: 20-30 parts of sodium hyaluronate, 20-30 parts of glycerol, 8-15 parts of carbomer, 3-8 parts of olive oil, 1-5 parts of dipotassium glycyrrhizinate, and 900+n parts of water; wherein n is 10 times the number of dipotassium glycyrrhizinate.

[0018] Preferably, the hyaluronic acid matrix is ​​composed of the following ingredients: 20-25 parts of sodium hyaluronate, 25-30 parts of glycerol, 10-12 parts of carbomer, 4-6 parts of olive oil, 3-5 parts of dipotassium glycyrrhizinate, and 900+n parts of water; wherein n is 10 times the number of parts of dipotassium glycyrrhizinate.

[0019] Further preferably, the hyaluronic acid matrix is ​​composed of the following ingredients: 20 parts of sodium hyaluronate, 30 parts of glycerol, 10 parts of carbomer, 5 parts of olive oil, 3 parts of dipotassium glycyrrhizinate, and 930 parts of water.

[0020] The hyaluronic acid matrix is ​​in a gel state and has a certain product dynamic viscosity, such as a dynamic viscosity of 120-140 Pa.s, or a dynamic viscosity of 120-132 Pa.s.

[0021] The bioburden of the hyaluronic acid matrix is ​​approximately 0.1, and is suitable for irradiation doses close to that when the bioburden is 0.1, such as irradiation doses of 8.0-11.0 kGy.

[0022] In another aspect, the present invention provides a method for preparing the aforementioned hyaluronic acid matrix.

[0023] Specifically, the preparation method includes the following steps: heating 900 parts of purified water to 60-80°C, adding sodium hyaluronate, glycerin, and carbomer in sequence, stirring at a constant temperature, cooling to 40-45°C after stirring, adding olive oil, stirring at a constant temperature, and after leaving at room temperature, adding a dipotassium glycyrrhizinate solution dissolved in the remaining n parts of water to prepare a semi-finished product.

[0024] Preferably, the heating temperature is 70°C-75°C.

[0025] Preferably, the heating time is 15-50 min, preferably 40-50 min.

[0026] Preferably, the temperature is lowered to 45°C.

[0027] Preferably, the preparation method comprises the following steps: heating 900 parts of purified water to 75°C, adding sodium hyaluronate, glycerin and carbomer in sequence, stirring at this temperature for 30 minutes, cooling to 45°C after stirring, adding olive oil, stirring at this temperature for 10 minutes, leaving at room temperature, and adding dipotassium glycyrrhizinate solution dissolved in the remaining 30 parts of water to prepare a semi-finished product.

[0028] Specifically, the preparation method also includes irradiation sterilization.

[0029] Preferably, the irradiation sterilization condition is 8.0-11.0 kGy.

[0030] In yet another aspect, the present invention provides use of the aforementioned hyaluronic acid matrix in preparing a gel product.

[0031] Specifically, the gel product can be a medical device, a cosmetic, a medicine or a daily necessity.

[0032] When the gel product is a medical device:

[0033] The medical devices include but are not limited to medical external dressings and medical spreadable gels.

[0034] When the gel product is a cosmetic:

[0035] The cosmetics may also include fucoidan.

[0036] The cosmetics include, but are not limited to: one or more of: facial cream, facial mask, hand cream, body lotion, lip balm, eye cream, eye mask, hand mask, and foot mask.

[0037] All types of gel cosmetics applicable in the art are within the protection scope of the present invention, and are not limited to the types listed.

[0038] The cosmetics can be color cosmetics or skin care products.

[0039] The application parts of the cosmetics include but are not limited to the face or the body.

[0040] When the gel product is a medicine:

[0041] The medicine is generally an external medicine, and may also include other medically and pharmaceutically acceptable carriers or excipients, and may also include medicinal active ingredients.

[0042] When the gel product is a daily necessity:

[0043] The daily necessities can be daily care products, such as private parts gel.

[0044] In yet another aspect, the present invention provides a gel product.

[0045] The gel product can be a medical device, a cosmetic, a medicine or a daily necessity.

[0046] The gel product comprises the aforementioned hyaluronic acid matrix.

[0047] When the gel product is a medical device, it can be a medical dressing or a medical smearable gel, and can also include other medically and pharmaceutically acceptable carriers or excipients, and can also include medicinal active ingredients.

[0048] When the gel product is a cosmetic, it may also include other ingredients commonly used in the cosmetics field, such as one or more of whitening ingredients, anti-wrinkle ingredients, repairing ingredients, moisturizing ingredients, and oil-controlling ingredients.

[0049] When the gel product is a medicine, it is generally an external medicine, and may also include other medically and pharmaceutically acceptable carriers or excipients, and may also include medicinal active ingredients.

[0050] When the gel product is a daily necessity, it can be a product used for daily care, such as private parts gel.

[0051] Beneficial effects of the present invention:

[0052] The hyaluronic acid matrix provided by the present invention is a gel matrix obtained by adjusting the formula, optimizing the preparation method accordingly, and adjusting the irradiation sterilization method. The stability meets the requirements of 2-year quality assurance, and the salt resistance and stability of the gel matrix containing carbomer are improved. It can maintain a good dynamic viscosity under certain salt concentration conditions and can be better applied to production and clinical practice. DETAILED DESCRIPTION

[0053] The present invention will be further described in detail below in conjunction with specific examples. The following examples are not intended to limit the present invention, but are only intended to illustrate the present invention. The experimental methods used in the following examples are generally conventional, unless otherwise specified, and the materials, reagents, etc. used in the following examples are commercially available, unless otherwise specified.

[0054] In the following examples, the sterility test, dynamic viscosity, and pH value detection methods are all referred to the 2020 edition of the "Chinese Pharmacopoeia".

[0055] Example 1

[0056] Formula: 20 parts of sodium hyaluronate, 30 parts of glycerin, 10 parts of carbomer, 5 parts of olive oil, 3 parts of dipotassium glycyrrhizinate, and 930 parts of purified water.

[0057] Preparation method: Heat 900 parts of purified water to 75°C, add sodium hyaluronate, glycerin, and carbomer in turn, keep warm and stir for 30 minutes, cool to 45°C after stirring, add olive oil, keep warm and stir for 10 minutes, leave at room temperature, and add dipotassium glycyrrhizinate solution dissolved in the remaining 30 parts of water. Control the environmental conditions of the production workshop so that the bioburden of the semi-finished product is close to 0.1 when sampling.

[0058] After packaging, the product is sterilized by irradiation at 11.0 kGy. The product undergoes sterility inspection and meets the sterility requirements. The dynamic viscosity of the product is 120 Pa.s, which meets the requirements of gel agents.

[0059] Example 2

[0060] Formula: 25 parts of sodium hyaluronate, 25 parts of glycerin, 12 parts of carbomer, 4 parts of olive oil, 5 parts of dipotassium glycyrrhizinate, and 950 parts of purified water.

[0061] Preparation method: Heat 900 parts of purified water to 70°C, add sodium hyaluronate, glycerin, and carbomer in sequence, keep warm and stir for 35 minutes, cool to 45°C after stirring, add olive oil, keep warm and stir for 10 minutes; after leaving at room temperature, add dipotassium glycyrrhizinate solution dissolved in the remaining 50 parts of water. Control the environmental conditions of the production workshop so that the bioburden of the semi-finished product is close to 0.1 when sampling.

[0062] After packaging, the product is sterilized by irradiation at 11.0 kGy. The product undergoes sterility inspection and meets the sterility requirements. The dynamic viscosity of the product is 132 Pa.s, which meets the requirements of gel agents.

[0063] Example 3

[0064] Formula: 30 parts of sodium hyaluronate, 20 parts of glycerin, 10 parts of carbomer, 6 parts of olive oil, 4 parts of dipotassium glycyrrhizinate, and 940 parts of purified water.

[0065] Preparation method: Heat 900 parts of purified water to 75°C, add sodium hyaluronate, glycerin, and carbomer in sequence, keep warm and stir for 30 minutes, cool to 45°C after stirring, add olive oil, keep warm and stir for 10 minutes; after leaving at room temperature, add dipotassium glycyrrhizinate solution dissolved in the remaining 40 parts of water. Control the environmental conditions of the production workshop so that the bioburden of the semi-finished product is close to 0.1 when sampling.

[0066] After packaging, the product is sterilized by irradiation at 11.0 kGy. The product undergoes sterility inspection and meets the sterility requirements. The dynamic viscosity of the product is 126 Pa.s, which meets the requirements of gel agents.

[0067] Comparative Example 1-2

[0068] A comparative experiment was set up based on the sterilization conditions in Example 1-2 and product testing was performed. The specific conditions and results are as follows:

[0069] the difference Dynamic viscosity Pa.s Comparative Example 1 (Comparative Example 1) Irradiation sterilization 25kGy 1.67 Comparative Example 2 Irradiation sterilization 25kGy 1.16

[0070] The product meets the sterility requirements after sterility testing, but the dynamic viscosity matrix performance cannot meet clinical requirements.

[0071] Example 4 Stability Test Comparison

[0072] The stability of the gels prepared in Examples 1-3 was investigated.

[0073] Example 1 Accelerated aging study, temperature 50 ° C, humidity 65%, accelerated inspection for 15 weeks (equivalent to 104 weeks), proved that the performance stability of the matrix of the present invention meets the requirements of 2-year quality assurance:

[0074]

[0075] Example 2 Accelerated aging study, temperature 50 ° C, humidity 65%, accelerated inspection for 15 weeks (equivalent to 104 weeks), proved that the performance stability of the matrix sterilized by the process of the present invention meets the requirement of 2 years:

[0076]

[0077] Example 3 Accelerated aging study, temperature 50 ° C, humidity 65%, accelerated inspection for 15 weeks (equivalent to 104 weeks), proved that the performance stability of the matrix sterilized by the process of the present invention meets the requirement of 2 years:

[0078]

[0079] Example 5 Gel Matrix Performance

[0080] The gel matrix of Examples 1-3 includes carbomer, which mainly serves to adjust the viscosity, and the salt content has a great influence on the viscosity of carbomer. An increase in the amount of salt will cause the viscosity of carbomer to decrease and the solution to become turbid, affecting the product experience. The salt content of the components in the examples of the present application is not sufficient to affect the state and properties of the gel matrix.

[0081] However, the gel in the example is used as a matrix and needs to be combined with other active ingredients when used. Various plant extracts may be added in the prior art. These extracts contain salts from the plants themselves and may have residual salts in the extraction method, which often affect the stability of the gel, especially plant extracts from the ocean. Clinical trial results show that 0.3% (mass fraction) of fucoidan has a significant effect on skin whitening, indicating that fucoidan has a good whitening effect. This example aims to detect the salt tolerance of the gel matrix in Example 2 and its stability when compounded with marine plant extracts (fucoidan).

[0082] In this example, dipotassium glycyrrhizinate and sodium chloride in the original formula were selected as agents for salt tolerance. Fucoidan was purchased from Qingdao Mingyue Seaweed Bio-Health Technology Group Co., Ltd., with a CAS number of 9072-19-9.

[0083] (1) Salt tolerance

[0084] 0.1 g, 0.5 g, 1 g, 1.5 g, 2 g, and 3 g of dipotassium glycyrrhizinate were added to the gel matrix (100 mL) of Examples 1-3, respectively. The dynamic viscosity was tested after 30 min. The results were as follows:

[0085]

[0086] It can be seen that the gel matrix of Examples 1-3 can maintain a good dynamic viscosity when an additional 15 g / L (about 1.5% V / V) of dipotassium glycyrrhizinate is added.

[0087] Considering that the addition amount of conventional dipotassium glycyrrhizinate is not too high, this embodiment also performs a dynamic viscosity test using sodium chloride as an indicator:

[0088] 0.1 g, 0.5 g, 1 g, 1.5 g, 2 g, and 3 g of sodium chloride were added to the gel matrix (100 mL) of Examples 1-3, respectively. The dynamic viscosity was measured after 30 min. The results were as follows:

[0089]

[0090] It can be seen that the gel matrix of Examples 1-3 can maintain good dynamic viscosity when an additional 15 g / L (about 1.5% V / V) of sodium chloride is added.

[0091] (2) Effect of fucoidan dosage

[0092] 0.1 g, 0.5 g, 1 g, 1.5 g, 2 g, and 3 g of fucoidan were added to the gel matrix (100 mL) of Examples 1-3, respectively. The dynamic viscosity was tested after 30 min. The results were as follows:

[0093]

[0094]

[0095] Since fucoidan itself has a certain function of regulating the dynamic viscosity of the gel, the dynamic viscosity of the gel matrix increased slightly when a small amount of fucoidan was added, but as the amount of fucoidan added increased, the dynamic viscosity of the gel matrix gradually decreased. The results showed that the gel matrix of Examples 1-3 could maintain a dynamic viscosity basically consistent with that of the original gel when the amount of fucoidan added was 20 g / L (about 2% V / V), and could still maintain a qualified gel dynamic viscosity when the amount of fucoidan added was 30 g / L (about 3% V / V).

[0096] (3) Stability study after adding fucoidan

[0097] According to the above results, 20 g / L (about 2% V / V) of fucoidan was added to the gel matrix of Examples 1-3, and an accelerated aging study was performed according to the method of Example 4 to observe the dynamic viscosity stability after the addition of fucoidan. The results are as follows:

[0098] 0 Weeks 3 weeks 6 weeks 9 weeks 15 weeks Example 1 Dynamic viscosity / Pa.s 132 132 133 131 131 Example 2 Dynamic viscosity / Pa.s 144 142 140 145 140 Example 3 Dynamic viscosity / Pa.s 151 150 152 154 150

[0099] The results showed that the gel matrix with fucoidan added could still maintain a good dynamic viscosity for a certain period of time.

[0100] Comparative Examples 3-5

[0101] A comparative example was set up with reference to the preparation method of Example 1, with the following specific differences:

[0102] Differences from Example 1 Comparative Example 3 No olive oil added Comparative Example 4 No added dipotassium glycyrrhizinate Comparative Example 5 No added olive oil and dipotassium glycyrrhizinate

[0103] Referring to the method of Example 4, the salt tolerance of the gel matrix prepared in Comparative Examples 3-5 and the stability when compounded with brown algae polysaccharide were tested, and the results are as follows:

[0104]

[0105]

[0106] Note: The salt tolerance (sodium chloride) concentration V / V refers to the highest salt concentration at which the dynamic viscosity is consistent with the gel matrix prepared in Example 1 (±5); The number of stable days after the addition of fucoidan refers to the longest number of weeks at which the dynamic viscosity is consistent with the gel matrix prepared in Example 1 (±5) when the concentration of fucoidan added is 2% V / V.

Claims

1. A hyaluronic acid matrix that can be sterilized by irradiation, characterized in that It is composed of the following ingredients: 20-30 parts of sodium hyaluronate, 20-30 parts of glycerin, 8-15 parts of carbomer, 3-8 parts of olive oil, 1-5 parts of dipotassium glycyrrhizinate, and 900+n parts of water; wherein n is 10 times the number of parts of dipotassium glycyrrhizinate; The preparation method of the hyaluronic acid matrix comprises the following steps: heating 900 parts of purified water to 70-75° C., adding sodium hyaluronate, glycerin, and carbomer in sequence, keeping warm and stirring, cooling to 40-45° C. after stirring, adding olive oil, keeping warm and stirring, leaving at room temperature, and adding a dipotassium glycyrrhizinate solution dissolved in the remaining n parts of water to prepare a semi-finished product; The preparation method also includes irradiation sterilization, and the irradiation sterilization condition is 11.0 kGy.

2. The hyaluronic acid matrix according to claim 1, characterized in that It is composed of the following ingredients: 20-25 parts of sodium hyaluronate, 25-30 parts of glycerol, 10-12 parts of carbomer, 4-6 parts of olive oil, 3-5 parts of dipotassium glycyrrhizinate, and 900+n parts of water; wherein n is 10 times the number of parts of dipotassium glycyrrhizinate.

3. The method for preparing the hyaluronic acid matrix according to claim 1, characterized in that: The method comprises the following steps: heating 900 parts of purified water to 70-75°C, sequentially adding sodium hyaluronate, glycerin and carbomer, keeping warm and stirring, cooling to 40-45°C after stirring, adding olive oil, keeping warm and stirring, placing at room temperature, and adding dipotassium glycyrrhizinate solution dissolved in the remaining n parts of water to prepare a semi-finished product; the preparation method also comprises irradiation sterilization, and the irradiation sterilization condition is 11.0 kGy.

4. The preparation method according to claim 3, characterized in that: The method comprises the following steps: heating 900 parts of purified water to 75°C, adding sodium hyaluronate, glycerin and carbomer in sequence, stirring at the temperature for 30 minutes, cooling to 45°C after stirring, adding olive oil, stirring at the temperature for 10 minutes, placing at room temperature, and adding a dipotassium glycyrrhizinate solution dissolved in the remaining 30 parts of water to prepare a semi-finished product.

5. Use of the hyaluronic acid matrix according to any one of claims 1 to 2 in the preparation of a gel.

6. The use according to claim 5, characterized in that: The gel is used for medical devices, cosmetics or medicines.

7. The use according to claim 6, characterized in that: The gel also includes fucoidan.

8. The use according to claim 6, characterized in that: The medical device is a medical external dressing.

9. The use according to claim 6, characterized in that: The medical device is a medical spreadable gel.

10. A gelling agent, characterized in that: The invention comprises the hyaluronic acid matrix according to any one of claims 1 to 2.

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