Product containing flexible glucan, preparation method, composition and application

Abstract

The invention relates to the technical field of cosmetics, in particular to a product containing flexible glucan, which can be prepared by treating materials containing glucan by adopting high-pressure microjet. According to the product containing the flexible dextran, the flexible dextran has a special intramolecular and intermolecular arrangement mode, has excellent effects of promoting wound healing and resisting inflammation, has an outstanding effect of promoting skin repair after being added into cosmetics, can effectively improve the after-healing effect of dot matrix laser, acid brushing, microneedle and other medical beauty projects, and has a good application prospect. Meanwhile, the repair of sensitive skin can be promoted. According to the preparation method of the product containing the flexible glucan, the intramolecular and intermolecular arrangement form of the glucan can be adjusted, the flexibility of the product is improved, meanwhile, the room-temperature stability of the product is improved, and the product can be massively added into cosmetics.

Description

Technical Field

[0001] This invention relates to the field of skincare technology, and more particularly to a product containing flexible dextran and its preparation and application. Background Art

[0002] Due to its polyhydroxy structure and long linear chain, dextran undergoes intramolecular and intermolecular bonding during storage, leading to aggregation and clustering of dextran molecules. On one hand, this aggregation blocks numerous active sites, reducing biological activity; on the other hand, intermolecular aggregation causes product instability.

[0003] In order to improve the stability of dextran, related technologies have adopted methods such as changing functional groups and removing macromolecular dextran in the preparation process. These methods essentially change the composition of dextran products, while also making the preparation process more complex and increasing production costs.

[0004] In view of this, the present invention is hereby proposed. Summary of the Invention

[0005] One object of the present invention is to provide a product containing flexible dextran that can solve the problems of decreased efficacy and stability of such products during long-term storage.

[0006] Another object of the present invention is to provide a method for preparing a product containing flexible dextran.

[0007] Another object of the present invention is to provide a composition comprising the above-mentioned product containing flexible dextran.

[0008] Another object of the present invention is to provide the use of the above-mentioned product / composition containing flexible dextran in the preparation of pharmaceutical products and / or cosmetics.

[0009] To achieve the above-mentioned objectives of the present invention, one aspect of the present invention provides a product containing flexible dextran, wherein the flexible dextran molecules in the product have a globular chain structure and a molecular conformation parameter of 0.5-0.8.

[0010] This invention contains a flexible dextran product, in which the dextran has a special intramolecular and intermolecular arrangement, exhibiting excellent wound healing and anti-inflammatory effects. When added to cosmetics, it has a prominent effect on promoting skin repair and can effectively improve the post-treatment effects of medical aesthetic procedures such as fractional laser, chemical peels, and microneedling, while also promoting the repair of sensitive skin.

[0011] Another aspect of the present invention provides a method for preparing a product containing flexible dextran, comprising treating the material containing dextran with a high-pressure microjet.

[0012] The preparation method of the product containing flexible dextran of the present invention can adjust the arrangement of dextran molecules within and between molecules, improve the flexibility of the product and increase its room temperature stability, so that it can be added in large quantities in cosmetics.

[0013] In one specific embodiment of the present invention, the dextran includes β-glucan.

[0014] In one specific embodiment of the present invention, the dextran includes cereal β-glucan.

[0015] In one specific embodiment of the present invention, the dextran includes one or more of oat β-glucan, barley β-glucan, and barley β-glucan.

[0016] In one specific embodiment of the present invention, the material containing dextran includes an extract containing dextran.

[0017] In one specific embodiment of the present invention, during the process of treating the material containing dextran with high-pressure microjet, the mass ratio of the material containing dextran to the fluid is 1:(12-70), and more preferably 1:14.

[0018] In one specific embodiment of the present invention, the fluid includes water.

[0019] In one specific embodiment of the present invention, the high-pressure microjet pressure is a gauge pressure of 80-140 MPa, and more preferably a gauge pressure of 120 MPa.

[0020] In one specific embodiment of the present invention, the high-pressure microjet is applied 1-5 times, more preferably 2 times.

[0021] In one specific embodiment of the present invention, the high-pressure microjet feed temperature is 0-45°C; more preferably 25°C.

[0022] In one specific embodiment of the present invention, the material containing dextran is heated and dissolved, and then subjected to high-pressure microjet treatment.

[0023] In one specific embodiment of the present invention, the heating and dissolving temperature is 70-90°C; more preferably, the heating and dissolving temperature is 80-85°C.

[0024] In one specific embodiment of the present invention, the preparation method further includes at least one of decolorization, desalting, and protein removal.

[0025] In one specific embodiment of the present invention, the decolorization is performed 1-3 times, preferably 2 times.

[0026] In one specific embodiment of the present invention, the total amount of decolorizing agent added is 2%-5% of the total mass of the material to be decolorized, more preferably 2-3%.

[0027] In one specific embodiment of the present invention, the decolorizing agent includes activated carbon.

[0028] In one specific embodiment of the present invention, the decolorization time for each decolorization is 30-60 minutes.

[0029] In one specific embodiment of the present invention, the temperature for each decolorization is 75-85°C.

[0030] In one specific embodiment of the present invention, the decolorization is performed twice. The first time, 3% of the total mass of the material to be decolorized is added as a decolorizing agent, and the decolorization is carried out at 80-85°C for 30 minutes. The second time, 2% of the total mass of the material to be decolorized is added as a decolorizing agent, activated carbon, and the decolorization is carried out at 80-85°C for 30 minutes.

[0031] In one specific embodiment of the present invention, anion and cation exchange resins are used for desalination.

[0032] In one specific embodiment of the present invention, the material flow rate during the desalination process is 1-3 t / h, and more preferably, the material flow rate during the desalination process is 2-3 t / h.

[0033] In one specific embodiment of the present invention, the obtained product is filtered and then compounded to obtain a product containing flexible dextran.

[0034] In one specific embodiment of the present invention, one or more of hexanediol, pentanediol, glycerol and PHEG are used as compounding solvents for compounding.

[0035] In one specific embodiment of the present invention, the compounding process is followed by sterilization.

[0036] In another aspect, the present invention provides a composition containing flexible dextran, comprising the above-mentioned product containing flexible dextran, and / or comprising a product containing flexible dextran prepared by the above-mentioned method for preparing the product containing flexible dextran.

[0037] In another aspect, the present invention provides the use of the above-mentioned product containing flexible dextran, the product containing flexible dextran prepared by the above-mentioned method for preparing the above-mentioned product containing flexible dextran, or the above-mentioned composition containing flexible dextran in the preparation of pharmaceutical products and / or cosmetics.

[0038] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0039] This invention relates to a product containing flexible dextran, in which the dextran has a special intramolecular and intermolecular arrangement, exhibiting excellent wound healing and anti-inflammatory effects. When added to cosmetics, it has a prominent effect on promoting skin repair and can effectively improve the post-treatment effects of medical aesthetic procedures such as fractional laser, chemical peels, and microneedling, while also promoting the repair of sensitive skin.

[0040] The preparation method of the product containing flexible dextran of the present invention can adjust the arrangement of dextran molecules within and between molecules, improve the flexibility of the product and increase its room temperature stability, so that it can be added in large quantities in cosmetics. Attached Figure Description

[0041] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the specific embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0042] Figure 1 This is a scanning electron microscope image of a flexible dextran sample from Example 3 of the present invention, wherein... Figure 1 a and Figure 1 b are scanning electron microscope images of flexible dextran samples from Example 3 of the present invention at different proportions;

[0043] Figure 2 This is a scanning electron microscope image of the flexible dextran sample from Example 7 of the present invention, wherein... Figure 2 a and Figure 2 b are scanning electron microscope images of flexible dextran samples from Example 7 of the present invention at different proportions;

[0044] Figure 3 This is a scanning electron microscope image of the dextran sample from Comparative Example 1 of this invention, wherein... Figure 3 a and Figure 3 b are scanning electron microscope images of the dextran samples of Comparative Example 1 of the present invention at different proportions;

[0045] Figure 4 This is a scanning electron microscope image of the dextran sample from Comparative Example 2 of this invention, wherein... Figure 4 a and Figure 4 b are scanning electron microscope images of the dextran samples of Comparative Example 2 of the present invention at different proportions;

[0046] Figure 5 This is a scanning electron microscope image of the dextran sample from Comparative Example 3 of this invention, wherein... Figure 5 a and Figure 5 b are scanning electron microscope images of the dextran samples of Comparative Example 3 of the present invention at different proportions;

[0047] Figure 6 This is a scanning electron microscope image of the dextran sample from Comparative Example 4 of this invention, wherein... Figure 6 a and Figure 6 b are scanning electron microscope images of the dextran samples of Comparative Example 4 of the present invention at different proportions;

[0048] Figure 7 This is an atomic force microscope image of a flexible dextran sample from Example 3 of the present invention, wherein... Figure 7 a and Figure 7 b are atomic force microscope images of the flexible dextran samples of Example 3 of the present invention at different proportions;

[0049] Figure 8 This is an atomic force microscope image of the dextran sample in Comparative Example 2 of this invention, wherein... Figure 8 a and Figure 8 b are atomic force microscope images of the dextran samples of Comparative Example 2 of the present invention at different proportions;

[0050] Figure 9 Bright field image (100X) of scratch healing of keratinocytes (HaCaT) in a specific embodiment of the present invention;

[0051] Figure 10 This is a comparison chart of scratch healing rates of flexible dextran samples in a specific embodiment of the present invention;

[0052] Figure 11 This is a comparative data graph showing the characterization of the anti-inflammatory efficacy of flexible dextran samples in specific embodiments of the present invention. Detailed Implementation

[0053] The technical solution of the present invention will be clearly and completely described below with reference to the accompanying drawings and specific embodiments. However, those skilled in the art will understand that the embodiments described below are some embodiments of the present invention, but not all embodiments, and are only used to illustrate the present invention, and should not be regarded as limiting the scope of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall be followed. Where the manufacturers of reagents or instruments are not specified, they are all conventional products that can be purchased commercially.

[0054] The present invention provides a product containing flexible dextran, wherein the flexible dextran molecules in the product have a globular chain structure and a molecular conformation parameter of 0.5-0.8.

[0055] This product contains flexible dextran, which has a unique intramolecular and intermolecular arrangement that provides excellent wound healing and anti-inflammatory effects. When added to cosmetics, it significantly improves skin repair and can effectively enhance the post-treatment results of cosmetic procedures such as fractional laser, chemical peels, and microneedling. It can also promote the repair of sensitive skin.

[0056] Another aspect of the present invention provides a method for preparing a product containing flexible dextran, comprising treating the material containing dextran with a high-pressure microjet.

[0057] High-pressure microjets exert a series of combined effects on fluid mixtures, including intense shearing and high-speed impact, and are often used to break down / crush materials or to homogenize them. They primarily utilize a hydraulic pump to generate high pressure (typically above 80 MPa), dispersing the fluid within the impact chamber into two or more fine streams that then collide intensely at high speeds within a very small flow channel. During the impact, most of the energy is instantly converted, generating a huge pressure drop, thereby causing the liquid particles to be highly fragmented. This invention creatively employs high-pressure microjets for the flexible modification of dextran, and various high-pressure microjets can be selected for operation.

[0058] This method can adjust the intramolecular and intermolecular arrangement of dextran molecules, improving the product's flexibility while increasing its room temperature stability, allowing it to be added in large quantities to cosmetics.

[0059] In one specific embodiment of the present invention, the dextran includes β-glucan, a natural product that is a polysaccharide composed of linked glucose molecules and is found in various plants and fungi. β-glucan possesses many important biological activities and pharmacological effects and is widely used in medicine, health products, food, and other fields.

[0060] In one specific embodiment of the present invention, the dextran suitable for use in the present invention can be derived from different sources, including cereal β-glucan. All cereal β-glucans are unbranched, linear, non-starch polysaccharides composed of glucose linked by β-1,3 and β-1,4 glycosidic bonds. Therefore, different cereal β-glucans have similar properties, namely, they all exhibit instability due to hydrogen bonding.

[0061] In one specific embodiment of the present invention, the dextran includes one or more of oat β-glucan, barley β-glucan, and barley β-glucan.

[0062] In one specific embodiment of the present invention, the material containing dextran includes a dextran extract. The dextran can be obtained from various sources through conventional extraction methods, or it can be purchased directly.

[0063] In one specific embodiment of the present invention, during the process of treating the material containing dextran with high-pressure microjet, the mass ratio of the material containing dextran to the fluid is 1:(12-70), and more preferably 1:14.

[0064] High-pressure microjet processing can be applied to materials containing dextran at different concentrations, depending on the actual equipment and the required concentration of the target product. Within the aforementioned mass ratio range of dextran-containing material to fluid, the requirements for flexible modification of dextran can be met. For example, in different embodiments, the mass ratio of dextran-containing material to fluid can be exemplarily 1:12, 1:12.5, 1:14, 1:18, 1:20, 1:26, 1:34, 1:37, 1:42, 1:50, 1:53, 1:62, 1:69, 1:70, etc.

[0065] In one specific embodiment of the present invention, the fluid includes water.

[0066] In one specific embodiment of the present invention, the high-pressure microjet pressure is a gauge pressure of 80-140 MPa, and more preferably a gauge pressure of 120 MPa.

[0067] In different embodiments, the high-pressure microjet pressure can be, for example, 80MPa, 85MPa, 90MPa, 102MPa, 114MPa, 120MPa, 131MPa, 140MPa, etc.

[0068] In one specific embodiment of the present invention, the high-pressure microjet is applied 1-6 times, more preferably 2 times.

[0069] In different implementations, the number of high-pressure microjet flows can be 1, 2, 3, 4, 5, or 6.

[0070] In one specific embodiment of the present invention, the high-pressure microjet feed temperature is 0-45°C; more preferably 25°C.

[0071] The feed temperature of the high-pressure microjets can be adjusted according to the usage requirements of the relevant equipment or the design needs of the actual production line. There are no strict temperature control requirements; generally, room temperature is sufficient. The flexible modification requirements for dextran can be met within the aforementioned high-pressure microjets feed temperature range. For example, in different embodiments, the high-pressure microjets feed temperature can be 0℃, 4℃, 8℃, 15℃, 20℃, 26℃, 32℃, 37℃, 41℃, 45℃, etc. In specific embodiments of this invention, the absence of a specific feed temperature indicates that the material temperature in the previous step or room temperature is sufficient for feeding.

[0072] In one specific embodiment of the present invention, the material containing dextran is heated and dissolved before being subjected to high-pressure microjet treatment. For general dextran materials, heating is a convenient operation to dissolve them, which facilitates the full utilization of the subsequent high-pressure microjet treatment.

[0073] In one specific embodiment of the present invention, the heating and dissolving temperature is 70-90°C; more preferably, the heating and dissolving temperature is 80-85°C.

[0074] Heating can improve the solubility of dextran. The ideal temperature is one that allows the dextran to dissolve completely; there are no strictly defined temperature requirements. The flexible modification needs of dextran can be met within the aforementioned temperature range. For example, in different embodiments, the heating and dissolving temperature can be exemplarily 70-73℃, 74-76℃, 78-80℃, 81-83℃, 88-90℃, etc.

[0075] In one specific embodiment of the present invention, the preparation method further includes at least one of decolorization, desalting, and protein removal. Other appropriate processing methods can be selected according to actual needs, such as the requirements of subsequent formulation applications.

[0076] In one specific embodiment of the present invention, the decolorization is performed 1-3 times, preferably 2 times.

[0077] In one specific embodiment of the present invention, the total amount of decolorizing agent added is 2%-5% of the total mass of the material to be decolorized, more preferably 2-3%.

[0078] The total amount of decolorizing agent added can be determined according to the color of the obtained product and the decolorization requirements. The use of conventional decolorizing agents does not affect the flexible properties of the dextran that has been treated. For example, in different embodiments, the total amount of decolorizing agent added can be 2%, 2.1%, 2.4%, 2.9%, 3.15%, 3.6%, 4.2%, 4.74%, 5% of the total mass of the material to be decolorized.

[0079] In one specific embodiment of the present invention, the decolorizing agent includes activated carbon.

[0080] In one specific embodiment of the present invention, the decolorization time for each decolorization is 30-60 minutes.

[0081] In different implementations, the time for each decolorization cycle can be exemplarily 30 min, 34 min, 39 min, 43 min, 46 min, 52 min, 57 min, 60 min, etc.

[0082] In one specific embodiment of the present invention, the temperature for each decolorization is 75-85°C.

[0083] The decolorization temperature can be set according to the requirements of equipment, production line, decolorization efficiency, etc. The decolorization temperature does not affect the flexibility of the dextran that has been processed. For example, in different embodiments, the decolorization temperature for each decolorization can be 75-77℃, 76-78℃, 78-80℃, 78.5-81℃, 83-85℃, etc.

[0084] In one specific embodiment of the present invention, the decolorization is performed twice. The first time, 3% of the total mass of the material to be decolorized is added as a decolorizing agent, and the decolorization is carried out at 80-85°C for 30 minutes. The second time, 2% of the total mass of the material to be decolorized is added as a decolorizing agent, activated carbon, and the decolorization is carried out at 80-85°C for 30 minutes.

[0085] In one specific embodiment of the present invention, anion and cation exchange resins are used for desalination.

[0086] In one specific embodiment of the present invention, the material flow rate during the desalination process is 1-3 t / h, and more preferably, the material flow rate during the desalination process is 2-3 t / h.

[0087] The material flow rate during the desalination process can be set according to the requirements of equipment, production line, desalination efficiency, etc. The material flow rate does not affect the flexible properties of the dextran that has been processed. For example, in different embodiments, the material flow rate during the desalination process can be 1-1.3t / h, 1.2-1.6t / h, 1.9-2.4t / h, 2-2.5t / h, 2.3-2.7t / h, 2.8-3t / h, etc.

[0088] By combining isoelectric point treatment with heat treatment, trace amounts of protein in the sample can be destroyed, denatured, and precipitated, then removed by subsequent filtration steps, thereby improving the appearance and stability of the product as needed. This invention can also use other protein removal processes as alternatives, such as the Sevage method, TCA method, and protease method, depending on product and process requirements.

[0089] In one specific embodiment of the present invention, the obtained product is filtered and then compounded to obtain a product containing flexible dextran.

[0090] In one specific embodiment of the present invention, one or more of hexanediol, pentanediol, glycerol and PHEG are used as compounding solvents for compounding.

[0091] In one specific embodiment of the present invention, the amount of the compound solvent can be adjusted as needed. In a preferred specific embodiment of the present invention, the obtained flexible dextran is added to glycerol, hexanediol and pentanediol, and the glycerol content in the obtained product is 10% of the total mass, the hexanediol content is 2% of the total mass, and the pentanediol content is 2% of the total mass.

[0092] In one specific embodiment of the present invention, the compounding process is followed by sterilization.

[0093] In another aspect, the present invention provides a composition containing flexible dextran, comprising the above-mentioned product containing flexible dextran, and / or comprising a product containing flexible dextran prepared by the above-mentioned method for preparing the product containing flexible dextran.

[0094] In another aspect, the present invention provides the use of the above-mentioned product containing flexible dextran, the product containing flexible dextran prepared by the above-mentioned method for preparing the above-mentioned product containing flexible dextran, or the above-mentioned composition containing flexible dextran in the preparation of pharmaceutical products and / or cosmetics.

[0095] Example 1

[0096] The specific steps for preparing flexible oat β-glucan are as follows:

[0097] Dissolve 7 kg of oat β-glucan powder in 98 kg of water (material-to-liquid ratio of 1:14), heat to 83°C and keep warm for 30 min, then cool to 25°C to obtain solution A;

[0098] Solution A was subjected to high-pressure microjet flow at a gauge pressure of 120 MPa, and the cycle was repeated twice to obtain 60 kg of solution B.

[0099] Solution B was diluted with deionized water at a mass ratio of solution B:water = 1:2 to obtain solution C;

[0100] Solution C was stirred and heated to 75°C. 3% of activated carbon by mass of solution C was added. The mixture was heated to 83°C and kept at that temperature for 30 minutes. After settling for 12-18 hours, the supernatant was extracted to obtain solution D.

[0101] Solution D was heated to 75°C, and 2% (by mass) of activated carbon was added. The mixture was kept at 84.8°C for 30 minutes, then cooled to 62°C and filtered to obtain solution E.

[0102] Solution E was desalted by passing it through an anion and cation exchange column at a flow rate of 3.0 TPH, and then the pH was adjusted to 4.64 to obtain solution F;

[0103] Add 1% (by mass) of activated carbon to solution F, then heat to 92 degrees Celsius and keep warm for 20 minutes, then filter to obtain flexible oat β-glucan;

[0104] Flexible oat β-glucan was added to glycerol, hexanediol and pentanediol. The resulting product contained 10% glycerol, 2% hexanediol and 2% pentanediol by mass. It was then sterilized at 100°C for 30 min.

[0105] Example 2

[0106] The specific steps for preparing flexible oat β-glucan are as follows:

[0107] 20 kg of commercially available oat β-glucan aqueous solution with a mass fraction of 1% (purchased from Beijing Dongfang Miaosen Biotechnology Co., Ltd., which also contains 12% glycerol and 1% PEHG) was subjected to high-pressure micro-jet at a gauge pressure of 120 MPa and circulated once to obtain 20 kg of flexible oat β-glucan.

[0108] Example 3

[0109] The specific steps for preparing flexible oat β-glucan are as follows:

[0110] 20 kg of commercially available oat β-glucan aqueous solution with a mass fraction of 1% (purchased from Beijing Dongfang Miaosen Biotechnology Co., Ltd., which also contains 12% glycerol and 1% PHEG) was subjected to high-pressure micro-jet at a gauge pressure of 120 MPa and circulated twice to obtain 20 kg of flexible oat β-glucan.

[0111] Example 4

[0112] The specific steps for preparing flexible oat β-glucan are as follows:

[0113] 20 kg of commercially available oat β-glucan aqueous solution with a mass fraction of 1% (purchased from Beijing Dongfang Miaosen Biotechnology Co., Ltd., which also contains 12% glycerol and 1% PHEG) was subjected to high-pressure micro-jet at a gauge pressure of 120 MPa and circulated 3 times to obtain 20 kg of flexible oat β-glucan.

[0114] Example 5

[0115] The specific steps for preparing flexible oat β-glucan are as follows:

[0116] 20 kg of commercially available oat β-glucan aqueous solution with a mass fraction of 1% (purchased from Beijing Dongfang Miaosen Biotechnology Co., Ltd., which also contains 12% glycerol and 1% PHEG) was subjected to high-pressure micro-jet at a gauge pressure of 120 MPa and circulated 5 times to obtain 20 kg of flexible oat β-glucan.

[0117] Example 6

[0118] The specific steps for preparing flexible oat β-glucan are as follows:

[0119] 20 kg of commercially available oat β-glucan aqueous solution with a mass fraction of 1% (purchased from Beijing Dongfang Miaosen Biotechnology Co., Ltd., which also contains 12% glycerol and 1% PHEG) was subjected to high-pressure micro-jet at a gauge pressure of 120 MPa and circulated 5 times to obtain 20 kg of flexible oat β-glucan.

[0120] Example 7

[0121] The specific steps for preparing flexible oat β-glucan are as follows:

[0122] 20 kg of commercially available oat β-glucan aqueous solution with a mass fraction of 1% (purchased from Beijing Dongfang Miaosen Biotechnology Co., Ltd., which also contains a mass fraction of 12% glycerol and 1% PHEG) was subjected to high-pressure micro-jet at a gauge pressure of 120 MPa and circulated 6 times to obtain 20 kg of flexible oat β-glucan.

[0123] Example 8

[0124] The specific steps for preparing flexible oat β-glucan are as follows:

[0125] 5.1 kg of oat β-glucan powder (purchased from Beijing Dongfang Miaosen Biotechnology Co., Ltd.) was dissolved in 71.4 kg of water, heated to 85°C and kept at that temperature for 30 min, then cooled to 25°C to obtain solution A;

[0126] Solution A was subjected to high-pressure microjet flow at a gauge pressure of 120 MPa, and the cycle was repeated twice to obtain 60 kg of solution B.

[0127] Solution B was diluted with deionized water at a mass ratio of solution B:water = 1:2 to obtain solution C;

[0128] Then add 5% (by mass) activated carbon to solution C, decolorize at 82℃ for 30 min, and cool and filter to obtain solution D;

[0129] Solution D was desalted by passing it through an anion and cation exchange column at a flow rate of 3.0 TPH, and then the pH was adjusted to 5 to obtain solution E;

[0130] Solution E was further cooled to 30°C and filtered to obtain flexible oat β-glucan;

[0131] Flexible oat β-glucan was added to glycerol, hexanediol and pentanediol. The resulting product contained 10% glycerol, 2% hexanediol and 2% pentanediol by mass. It was then sterilized at 100°C for 30 min.

[0132] Example 9

[0133] The specific steps for preparing flexible oat β-glucan are as follows:

[0134] 5.1 kg of oat β-glucan powder (purchased from Beijing Dongfang Miaosen Biotechnology Co., Ltd.) was dissolved in 71.4 kg of water, heated to 85°C and kept warm for 30 min, and then cooled to 25°C to obtain solution A;

[0135] Solution A was subjected to high-pressure microjet flow at a gauge pressure of 120 MPa, and the cycle was repeated twice to obtain 60 kg of solution B.

[0136] Solution B was diluted with deionized water at a mass ratio of solution B:water = 1:2 to obtain solution C;

[0137] Then add 3% (by mass) activated carbon to solution C, decolorize at 82℃ for 30 min, and cool and filter to obtain solution D;

[0138] Add 2% (by mass) of activated carbon to solution D, decolorize at 85°C for 30 min, then cool and filter to obtain solution E;

[0139] Solution E was desalted by passing it through an anion and cation exchange column at a flow rate of 3.0 TPH, and then the pH was adjusted to 5 to obtain solution F;

[0140] Solution F was further cooled to 30°C and filtered to obtain flexible oat β-glucan;

[0141] Flexible oat β-glucan was added to glycerol, hexanediol and pentanediol. The resulting product contained 10% glycerol, 2% hexanediol and 2% pentanediol by mass. It was then sterilized at 100°C for 30 min.

[0142] Example 10

[0143] The specific steps for preparing flexible oat β-glucan from oat β-glucan powder are as follows:

[0144] Oat β-glucan powder (purchased from Beijing Dongfang Miaosen Biotechnology Co., Ltd.) was dissolved in water at a mass ratio of oat β-glucan powder: water = 1:12. The solution was heated to 83°C and kept at that temperature for 30 minutes. The solution was then cooled to 25°C to obtain solution A.

[0145] Solution A was subjected to high-pressure microjet flow at a gauge pressure of 80 MPa, and the cycle was repeated twice to obtain solution B;

[0146] Solution B was diluted with water to a mass fraction of 1% for oat β-glucan to obtain solution C;

[0147] Solution C was stirred and heated to 75°C. 3% of activated carbon by mass of solution C was added. The mixture was heated to 83°C and kept at that temperature for 30 minutes. After settling for 12-18 hours, the supernatant was extracted to obtain solution D.

[0148] Solution D was heated to 75°C, and 2% (by mass) of activated carbon was added. The mixture was kept at 84.8°C for 30 minutes, then cooled to 62°C and filtered to obtain solution E.

[0149] Solution E was desalted by passing it through an anion and cation exchange column at a flow rate of 3.0 TPH, and then the pH was adjusted to 4.64 to obtain solution F;

[0150] Add 1% (by mass) of activated carbon to solution F, then heat to 92 degrees Celsius and keep warm for 20 minutes, then filter to obtain flexible oat β-glucan;

[0151] Flexible oat β-glucan was added to glycerol, hexanediol and pentanediol. The resulting product contained 10% glycerol, 2% hexanediol and 2% pentanediol by mass. It was then sterilized at 100°C for 30 min.

[0152] Example 11

[0153] The specific steps for preparing flexible oat β-glucan from oat β-glucan powder are as follows:

[0154] Oat β-glucan powder (purchased from Beijing Dongfang Miaosen Biotechnology Co., Ltd.) was dissolved in water at a mass ratio of oat β-glucan powder: water = 1:20. The solution was heated to 83°C and kept at that temperature for 30 minutes. The solution was then cooled to 25°C to obtain solution A.

[0155] Solution A was subjected to high-pressure microjet flow at a gauge pressure of 100 MPa, and the cycle was repeated twice to obtain solution B;

[0156] Solution B was diluted with water to a mass fraction of 1% for oat β-glucan to obtain solution C;

[0157] Solution C was stirred and heated to 75°C. 3% of activated carbon by mass of solution C was added. The mixture was heated to 83°C and kept at that temperature for 30 minutes. After settling for 12-18 hours, the supernatant was extracted to obtain solution D.

[0158] Solution D was heated to 75°C, and 2% (by mass) of activated carbon was added. The mixture was kept at 84.8°C for 30 minutes, then cooled to 62°C and filtered to obtain solution E.

[0159] Solution E was desalted by passing it through an anion and cation exchange column at a flow rate of 3.0 TPH, and then the pH was adjusted to 4.64 to obtain solution F;

[0160] Add 1% activated carbon by mass of solution F to solution F, then heat to 92 degrees Celsius and keep warm for 20 minutes, then filter to obtain flexible oat β-glucan;

[0161] Flexible oat β-glucan was added to glycerol, hexanediol and pentanediol. The resulting product contained 10% glycerol, 2% hexanediol and 2% pentanediol by mass. It was then sterilized at 100°C for 30 min.

[0162] Comparative Example 1

[0163] The specific steps for preparing oat β-glucan from oat β-glucan powder are as follows:

[0164] 7 kg of oat β-glucan powder (purchased from Beijing Dongfang Miaosen Biotechnology Co., Ltd.) was dissolved in 98 kg of water (material-to-liquid ratio of 1:14), heated to 83°C and kept warm for 30 min, and then cooled to 25°C to obtain solution A;

[0165] Solution A was diluted with deionized water at a mass ratio of solution A:water = 1:2 to obtain solution B;

[0166] Stir and heat solution B to 75°C, add 3% (by mass) of activated carbon from solution B, continue heating to 83°C and hold for 30 minutes, allow to settle for 12-18 hours, then extract the supernatant to obtain solution C.

[0167] Solution C was heated to 75°C, and 2% (by mass) of activated carbon was added. The mixture was kept at 84.8°C for 30 minutes, then cooled to 62°C and filtered to obtain solution D.

[0168] Solution D was desalted by passing it through an anion and cation exchange column at a flow rate of 3.0 TPH, and then the pH was adjusted to 4.64 to obtain solution E;

[0169] Add 1% (by weight) of activated carbon to solution E, then heat to 92 degrees Celsius and keep warm for 20 minutes, then filter to obtain oat β-glucan;

[0170] Flexible oat β-glucan was added to glycerol, hexanediol and pentanediol. The resulting product contained 10% glycerol, 2% hexanediol and 2% pentanediol by mass. It was then sterilized at 100°C for 30 min.

[0171] Comparative Example 2

[0172] The specific steps for preparing oat β-glucan are as follows:

[0173] Comparative Example 2 was a 1% oat β-glucan aqueous solution (purchased from Beijing Dongfang Miaosen Biotechnology Co., Ltd., which also contained 12% glycerol and 1% PHEG) of cosmetic grade.

[0174] Comparative Example 3

[0175] The specific steps for preparing high-pressure homogenized oat β-glucan are as follows:

[0176] The 1% oat β-glucan aqueous solution obtained in Comparative Example 2 was subjected to high-pressure homogenization once at a gauge pressure of 20 MPa.

[0177] Comparative Example 4

[0178] The specific steps for preparing high-pressure homogenized oat β-glucan are as follows:

[0179] The 1% oat β-glucan aqueous solution obtained in Comparative Example 2 was subjected to high-pressure homogenization, and homogenized twice at a gauge pressure of 40 MPa.

[0180] Comparative Example 5

[0181] The specific steps for preparing high-pressure homogenized oat β-glucan are as follows:

[0182] The 1% oat β-glucan aqueous solution obtained in Comparative Example 2 was subjected to high-pressure shear homogenization at 15000 r / min for 20 min.

[0183] The physicochemical properties of the samples prepared in Examples 1-11 and Comparative Examples 1-5 were tested, and the results are shown in Table 1:

[0184] Physicochemical index testing methods:

[0185] Colorimetry: Colorimetry was measured using a platinum-cobalt colorimeter (HI96727). First, ultrapure water was used for zeroing, and then samples of each example / comparative product were added for colorimetry testing.

[0186] Turbidity: Turbidity was measured using an ISO portable turbidimeter (HI98713-01). Before use, the turbidity was calibrated using a standard turbidity sample. Then, samples of each example / comparative example product were added for turbidity testing.

[0187] Table 1 Physicochemical properties of the embodiments and comparative samples of the present invention

[0188]

[0189]

[0190] According to the requirements of the cosmetics industry, it is preferred that the raw materials be clear and colorless or nearly colorless, so as to minimize the impact on the appearance of the finished product in the formulation application. Therefore, a color of less than 90 and a turbidity of less than 5 are preferred. Therefore, some of the comparative samples in the above groups can be selected if only appearance is considered. However, even if the appearance of different dextran samples meets the standards or is almost the same, their stability and efficacy may still show significant differences in subsequent formulation applications.

[0191] The stability of the samples prepared in Examples 1-11 and Comparative Examples 1-5 was observed, and the results are shown in Table 2:

[0192] Physicochemical index testing methods:

[0193] The samples were placed in a dark environment at room temperature (25℃±2℃, 60%RH±10%RH) and observed under white light in a small high-definition photography studio (Chunying) every 30 days. If the samples showed precipitation or gelation, they were recorded as unstable. The number of stable days was then counted and recorded.

[0194] Table 2. Stability observation results of the embodiments and comparative sample of the present invention.

[0195] Serial Number Sample Name Stable number of days 1 Example 1 420 2 Example 2 360 3 Example 3 390 4 Example 4 360 5 Example 5 360 6 Example 6 360 7 Example 7 330 8 Example 8 300 9 Example 9 280 10 Example 10 200 11 Example 11 200 12 Comparative Example 1 30 13 Comparative Example 2 60 14 Comparative Example 3 60 15 Comparative Example 4 60 16 Comparative Example 5 60

[0196] As shown in Table 2, the stability observation results of the embodiments of the present invention are significantly better than those of the comparative examples. The stability observation results of the embodiments of the present invention refer to the actual number of days observed up to the application date. No unstable phenomena such as stratification, precipitation, turbidity, or discoloration occurred within the above-mentioned time range. This does not mean that the stability can only be maintained for the above-mentioned time; those skilled in the art should be able to infer that the flexible dextran product of the present application can maintain stability for a longer period. In contrast, the stability observation results of comparative examples 1-5 indicate that unstable phenomena such as stratification, precipitation, turbidity, and discoloration occurred within the above-mentioned time, which would affect their final efficacy and application. Therefore, the flexible dextran product of the present invention has better stability and has broader application prospects in formulations, such as its use in combination with other raw materials and increasing its dosage.

[0197] Scanning microscopy methods:

[0198] Samples from each embodiment and comparative example were freeze-dried (freeze dryer, Boyikang, FD-1A-80). The freeze-dried samples were then cut open, and the cut surfaces were adhered to the stage using conductive tape. Gold sputtering was performed every 60 seconds using an ion sputtering instrument (Hitachi, E-1010) at a vacuum of 5 Pa and a current of 15 mA. Each sample was sputtered twice to increase conductivity. Finally, the morphology of the samples was photographed and recorded at 300X and 1000X magnification using a scanning microscope (Hitachi, S-3400N) at 10 kW.

[0199] The morphology of the flexible dextran sample of this invention was characterized by scanning electron microscopy, and the results are as follows: Figure 1-6 As shown:

[0200] according to Figure 1-6It can be seen that the samples of Comparative Examples 1-4 have obvious sheet-like structures in their microstructure, while the samples of Examples 3 and 7 of the present invention show obvious chain-like structures and form flexible spatial structures. The adjacent dextran chains extend in an orderly manner and do not stack and accumulate into sheet-like structures.

[0201] Atomic force microscopy method:

[0202] Each example / comparative example was sampled and lyophilized, then prepared into a 1 mg / mL solution, and then diluted to 5 μg / mL. 10 μL of the diluted test solution was taken and deposited onto freshly cleaved mica, and dried in air at room temperature for 1.5 hours. Measurement parameters: Covered with a petri dish before AFM imaging, measurements were performed using a Picoscan atomic force microscope in magnetic alternating current (MAC) mode.

[0203] The solution conformation of the flexible dextran sample of this invention was characterized by atomic force microscopy, and the results are as follows: Figures 7-8 As shown:

[0204] according to Figures 7-8 As can be seen, the microscopic three-dimensional structure of the sample in Comparative Example 2 is a dispersed point-like structure, which forms a sheet-like structure after being stacked. In contrast, the microscopic three-dimensional structure of the sample in Example 3 of this invention has a richer three-dimensional structure, in which the dextran molecules have obvious spherical chain-like structures, including some spatial network structures. This may be due to the weak binding effect between the flexible dextran chains under some weak forces, such as intermolecular forces and hydrogen bonds. This further promotes the excellent stability of the flexible dextran of this invention. At the same time, such a spatial network structure can better maintain the flexibility of the dextran chains, making it easy to use and better exert its effects.

[0205] Cell scratch assay method:

[0206] HaCaT cells were seeded at the appropriate density of 5*102 5 Cells were seeded per well into 6-well plates and incubated overnight in a CO2 incubator. The next day, the cells were scratched in the center of each well using a pipette tip. After scratching, the cells were washed with PBS and photographed under a microscope. The medium was then replaced with high-glucose DMEM (gibio, C11965500BT) containing 0.25% (w / w) of the sample from the test example / comparative example, and incubated overnight in a CO2 cell culture incubator. After 24 hours, the medium was discarded, replaced with PBS, and photographed under a microscope. Finally, the scratch healing rate was calculated based on the area of ​​the cell-free region before and after 24 hours from the microscopic images, using the following formula:

[0207] Scratch healing rate = (Area before scratch - Area after scratch in 24 hours) / Area before scratch * 100%

[0208] The flexible dextran samples obtained in Examples 1-11 and Comparative Examples 1-5 of this invention were characterized by cell scratch assay efficacy, and the results are as follows: Figures 9-10 As shown:

[0209] according to Figures 9-10 It can be seen that the samples of Examples 1-11 of this invention have a comprehensive improvement in cell repair compared with Comparative Examples 1-5, and some of the sample examples have reached the level of EGF (epidermal growth factor) in cell repair. EGF has a variety of biological activities, which can induce a series of biochemical activities in cells, stimulate epidermal and epithelial cells, promote epidermal proliferation and keratinization, strongly promote cell division, enhance cell activity, promote metabolism, and promote the synthesis of hyaluronic acid and glycoproteins. EGF can also promote the healing of skin and mucous membrane wounds, prevent and treat ulcers, and has anti-inflammatory and analgesic effects. It is widely used in the treatment of burns, scalds, surgical wounds, mechanical injuries, skin ulcers, laser cosmetic procedures, etc., and can effectively inhibit the growth of acne and pimples, and protect the skin and mucous membranes from or minimize mechanical and chemical damage. Due to its excellent efficacy, EGF has a wide range of applications in wound care (such as burns, scalds, abrasions, etc.). However, the injection of EGF in the medical aesthetics industry has huge safety risks, which can lead to uncontrollable local skin tissue growth, resulting in the formation of nodules and lumps, and in severe cases, disfigurement. In response, China's National Medical Products Administration has listed EGF as a prohibited ingredient. The flexible dextran product provided in this application can be considered a safe, mild, and effective plant-derived alternative to EGF.

[0210] Cellular anti-inflammatory experimental methods:

[0211] RAW264.7 cells were stored at the appropriate cell density of 3*102 5 Cells were seeded per well in 24-well plates and incubated overnight in a CO2 cell culture incubator. The next day, samples of the test examples / comparative examples were dissolved in high-glucose DMEM medium (gibio, C11965500BT) to prepare test solutions containing 0.25% (w / v) of the sampled products from different examples and comparative examples, and incubated for 24 h. On the third day, 10 ng / mL of LPS was added to induce inflammation and incubated for 24 h. On the fourth day, the supernatant of each group was analyzed for inflammatory factors according to the TNF-α ELISA kit.

[0212] The cellular anti-inflammatory efficacy of the flexible dextran samples obtained in Examples 1-11 and Comparative Examples 1-5 of this invention was characterized, and the results are as follows: Figure 11 As shown:

[0213] according to Figure 11It can be seen that the samples of Examples 1-11 of the present invention have a comprehensive improvement in anti-inflammatory properties compared with Comparative Examples 1-5, and even reach / exceed the level of dexamethasone at the general dosage (mass fraction of 0.001%, higher dosages may cause cytotoxicity). The flexible dextran product of the present invention has significant advantages in anti-inflammatory properties.

[0214] Samples from each embodiment and comparative example were lyophilized, and the lyophilized powder was analyzed for monosaccharide composition. The remaining samples were prepared into 1 mg / mL solutions for flexible structure characterization. The specific methods are as follows:

[0215] Monosaccharide composition determination: Take a clean chromatographic vial, weigh an appropriate amount of sample, add 1 ml of 2M TFA acid solution, and heat at 121℃ for 2 hours. Purge with nitrogen and dry. Wash with 99.99% methanol, then dry again, repeating the methanol washing 2-3 times. Dissolve in sterile water, transfer to a chromatographic vial for analysis.

[0216] The chromatographic system used was the Thermo ICS 5000+ ion chromatography system (ICS 5000+, Thermo Fisher Scientific, USA), which used an electrochemical detector to analyze and detect monosaccharide components.

[0217] Using Dionex TM CarboPac TM PA20 (150*3.0mm, 10μm) liquid chromatography column; injection volume: 5μL. Mobile phase A (H2O), mobile phase B (0.1M NaOH), mobile phase C (0.1M NaOH, 0.2M NaAc), flow rate 0.5 mL / min; column temperature 30℃; elution gradient: 0 min A phase / B phase / C phase (95:5:0, V / V), 26 min A phase / B phase / C phase (85:5:10, V / V), 42 min A phase / B phase / C phase (85:5:10, V / V), 42.1 min A phase / B phase / C phase (60:0:40, V / V), 52 min A phase / B phase / C phase (60:40:0, V / V), 52.1 min A phase / B phase / C phase (95:5:0, V / V), 60 min A phase / B phase / C phase (95:5:0, V / V).

[0218] The monosaccharide composition of each group of samples was obtained by comparing the spectrum with that of the standard sample.

[0219] Characterization of flexible structures:

[0220] The prepared 1 mg / mL sample solutions of each example / comparative example were filtered through a 0.45 μm filter membrane for analysis. The chromatographic system used was a gel permeation chromatography-differential chromatography-multi-angle laser light scattering system. The liquid chromatography system was a U3000 (Thermo, USA), the differential detector was an Optilab T-rEX (Wyatttechnology, CA, USA), and the laser light scattering detector was a DAWN HELEOS II (Wyatttechnology, CA, USA). The chromatographic conditions were as follows: Ohpak SB-805HQ (300×8 mm) and Ohpak SB-803HQ (300×8 mm) gel size exclusion columns were used in series. The column temperature was 45 °C, the injection volume was 100 μL, the mobile phase was A (mass fraction of 0.02% NaN3, 0.1 M NaNO3), the flow rate was 0.6 mL / min, and the elution gradient was isocratic for 75 min. The chromatographic data were processed using ASTRA 6.1 software. Molecular configuration diagrams were obtained with molar mass (g / mol) as the abscissa and root mean square radius (nm) as the ordinate. The slope of the slope was used as a molecular conformation parameter. Molecules with a slope < 0.5 are mostly spherical, and those with a slope > 0.8 are mostly rigid rod-like structures. In the specific embodiments of this application, molecules with a slope ≥ 0.8 and ≥ 0.5 are mostly flexible random coil configurations.

[0221] The monosaccharide composition and flexible structure of the flexible dextran prepared in the embodiments and comparative examples of the present invention were characterized, and the results are shown in Tables 3 and 4:

[0222] Table 3. Results of monosaccharide composition detection in the embodiments and comparative examples of the present invention. As can be seen from Table 3, the monosaccharide composition results of each comparative example and each embodiment of this application are similar, proving that the modification method of this patent will not cause changes in the monosaccharide composition of the samples and has little impact on the primary structure of polysaccharides.

[0223] Table 4. Molecular conformation detection results of embodiments and comparative examples of the present invention.

[0224]

[0225]

[0226] As shown in Table 4, the molecular configuration parameter in the comparative example of this application is 0.27 < 0.5, indicating a spherical configuration. In contrast, the molecular configuration parameter in the examples of this application is 0.62, falling between 0.5 and 0.8, indicating a flexible random coil configuration. Furthermore, the molecular weights of the comparative example and the examples are not significantly different, demonstrating that this application effectively regulates the structure of the dextran molecule, transforming it from its original spherical configuration to a flexible random coil configuration, thus obtaining flexible dextran.

[0227] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the above embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A product containing flexible dextran, characterized in that, The product containing flexible dextran has a globular chain structure with a molecular conformation parameter of 0.5-0.

8.

2. The product containing flexible dextran according to claim 1, characterized in that, The dextran includes β-glucan; Preferably, the dextran comprises cereal β-glucan; Preferably, the dextran includes one or more of oat β-glucan, barley β-glucan, and barley β-glucan.

3. A method for preparing a product containing flexible dextran, characterized in that, This includes using high-pressure microjet to process materials containing dextran.

4. The method for preparing a product containing flexible dextran according to claim 3, characterized in that, The dextran includes β-glucan; Preferably, the dextran comprises cereal β-glucan; Preferably, the dextran includes one or more of oat β-glucan, barley β-glucan, and barley β-glucan; Preferably, the material containing dextran includes a dextran extract; Preferably, in the process of treating the material containing dextran with high-pressure microjet, the mass ratio of the material containing dextran to the fluid is 1:(12-70), and more preferably 1:14; Preferably, the fluid comprises water; Preferably, the high-pressure microjet pressure is 80-140 MPa, and more preferably 120 MPa. Preferably, the high-pressure microjet is applied 1-6 times, more preferably 2 times; Preferably, the high-pressure microjet feed temperature is 0-45℃; more preferably, it is 25℃.

5. The method for preparing a product containing flexible dextran according to claim 3, characterized in that, The material containing dextran is heated and dissolved, and then subjected to high-pressure microjet treatment. Preferably, the heating and melting temperature is 70-90℃; more preferably, the heating and melting temperature is 80-85℃.

6. The method for preparing a product containing flexible dextran according to claim 3, characterized in that, The preparation method further includes at least one of decolorization, desalting, and protein removal.

7. The method for preparing a product containing flexible dextran according to claim 6, characterized in that, The decolorization process is performed 1-3 times, preferably 2 times; Preferably, the total amount of decolorizing agent added is 2%-5% of the total mass of the material to be decolorized, and more preferably 2-3%. Preferably, the decolorizing agent comprises activated carbon; Preferably, the decolorization time for each decolorization is 30-60 minutes; Preferably, the temperature for each decolorization step is 75-85℃; More preferably, the decolorization is performed twice. The first time, 3% of the total mass of the material to be decolorized is added as a decolorizing agent, and the decolorization is carried out at 80-85℃ for 30 minutes. The second time, 2% of the total mass of the material to be decolorized is added as a decolorizing agent (activated carbon), and the decolorization is carried out at 80-85℃ for 30 minutes. Preferably, anion and cation exchange resins are used for desalination; Preferably, the material flow rate during the desalination process is 1-3 t / h, and more preferably, the material flow rate during the desalination process is 2-3 t / h.

8. A method for preparing a product containing flexible dextran according to claim 5 or 6, characterized in that, The obtained product was filtered and then compounded to obtain a product containing flexible dextran. Preferably, one or more of hexanediol, pentanediol, glycerol, and PHEG are used as the compounding solvent; Preferably, the mixture is sterilized after compounding.

9. A composition containing flexible dextran, characterized in that, The product contains a flexible dextran as described in any one of claims 1-2, and / or contains a product containing flexible dextran prepared by the preparation method of a flexible dextran product as described in any one of claims 3-8.

10. The use of a product containing flexible dextran as described in any one of claims 1-2, a product containing flexible dextran prepared by the preparation method of a product containing flexible dextran as described in any one of claims 3-8, or a composition containing flexible dextran as described in claim 9 in the preparation of pharmaceutical products and / or cosmetics.

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