Composition for promoting collagen generation based on sophora flower powder as well as preparation method and application of composition

Abstract

The invention relates to a composition for promoting collagen generation based on sophora flower powder as well as a preparation method and application of the composition, and belongs to the technical field of sophora flower extracts. A preparation method of the sophora flower powder extract comprises the following steps: taking sophora flower powder, adding water, boiling, cooling, adding a carbon source and aspergillus oryzae CICC 2066, fermenting, sterilizing, filtering, concentrating and freeze-drying to obtain the sophora flower powder extract. The sophora flower powder extract can be applied to food, can also be applied to cosmetics, and can effectively promote the generation of collagen and improve the skin elasticity and tightening effect.

Description

Technical Field

[0001] This invention relates to a composition based on Sophora japonica flower powder that promotes collagen production, its preparation method, and its application, belonging to the technical field of Sophora japonica flower extract. Background Technology

[0002] The main active components of Sophora japonica flowers are flavonoids (such as rutin and quercetin), phenolic acids (such as chlorogenic acid), and a small amount of triterpenoid saponins. These components all have strong antioxidant activity: flavonoids (especially quercetin) can scavenge reactive oxygen species (ROS) in the skin or tissues, reducing oxidative damage to fibroblasts—fibroblasts are the core cells for collagen synthesis, and impaired function directly leads to reduced collagen synthesis. The phenolic hydroxyl structure can chelate metal ions (such as Fe). 2+ Cu 2+ It inhibits lipid peroxidation mediated by metal ions, prevents abnormal peptide chain breakage or cross-linking of collagen, and maintains its structural stability.

[0003] The active ingredients in Sophora japonica flower extract can enhance the synthetic function of fibroblasts by intervening in cell signaling: Quercetin can activate the MAPK / ERK pathway and the TGF-β / Smad pathway; the former promotes fibroblast proliferation and increases the "cellular base" for collagen synthesis; the latter is a key regulatory pathway for collagen synthesis, which can upregulate the gene expression of type I and type III collagen (COL1A1, COL3A1) and accelerate peptide chain synthesis. Some triterpenoid saponins can increase the activity of proline hydroxylase: collagen maturation depends on the hydroxylation of proline (forming a stable triple helix structure), and enhanced enzyme activity can promote the hydroxylation reaction and reduce the accumulation of immature collagen. The content of collagen in the human body depends not only on synthesis but also on the degradation rate. Sophora japonica flower extract can protect synthesized collagen by inhibiting degradation enzymes. Among them, ultraviolet radiation, inflammation, and other stimuli can induce the overexpression of matrix metalloproteinases (MMPs, such as MMP-1 and MMP-3), which can specifically hydrolyze the peptide bonds of collagen. Flavonoids in Sophora japonica flowers can downregulate the gene transcription of MMPs, reduce their activity, and decrease collagen breakdown. Phenolic acids (such as chlorogenic acid) can inhibit the activity of elastase. Although this enzyme mainly degrades elastin, excessive activation can indirectly damage the collagen fiber network. Inhibiting its activity can maintain the stability of collagen tissue distribution.

[0004] The polysaccharides and amino acids in Sophora japonica flower extract can serve as nutritional precursors, supporting collagen synthesis. Polysaccharides can regulate the structure of the extracellular matrix (ECM) surrounding fibroblasts, providing a suitable spatial environment for collagen assembly (forming fiber bundles). Some free amino acids (such as glycine and proline) are basic building blocks of collagen and can directly participate in peptide chain synthesis, indirectly improving synthesis efficiency.

[0005] Fermentation can enhance the variety and abundance of active ingredients in plant extracts, thereby increasing their functional strength. Currently, there are relatively few studies applying fermentation to the extraction of Sophora japonica flowers to enhance collagen production; therefore, further research is needed to validate the effectiveness of this technology. Simultaneously, further research is required to improve the collagen production efficiency of Sophora japonica flower extract complexes. Summary of the Invention

[0006] To address the aforementioned technical problems, this invention proposes a composition based on Sophora japonica pollen that promotes collagen production, its preparation method, and its application.

[0007] On one hand, this invention provides a method for preparing Sophora japonica flower powder extract, comprising: taking Sophora japonica flower powder, boiling it with water and then cooling it, adding a carbon source and Aspergillus oryzae for fermentation, sterilizing, filtering, concentrating, and freeze-drying. The use of Aspergillus oryzae to ferment Sophora japonica flowers, compared to other bacteria, can effectively promote the effect of Sophora japonica flower extract on enhancing collagen synthesis, resulting in plump and vibrant skin.

[0008] Furthermore, the sophora flower pollen extract can be used in food and cosmetics.

[0009] Furthermore, the sophora flower pollen extract can effectively promote collagen production, increasing skin elasticity and firmness.

[0010] Furthermore, in the preparation method of Sophora japonica flower powder extract, the carbon source includes substances such as sucrose, glucose, and fructose that can provide energy for microbial fermentation. Aspergillus oryzae (CICC 2066) can be selected as the *Aspergillus oryzae*, which can effectively and significantly enhance the effect of Sophora japonica flower extract in promoting the synthesis of type I and III collagen. The amount of carbon source can be 0.01-3% of the weight of the Sophora japonica flower powder, and the amount of *Aspergillus oryzae* used for inoculation fermentation is 10... 6 -10 11 One bacterial cell / g of Sophora japonica pollen. Fermentation temperature and time can be selected at 25-40℃ for 3-5 days.

[0011] The present invention also provides a composition based on Sophora japonica powder to promote collagen production, the raw materials of which are: Sophora japonica powder extract, polypeptide, Cynara scolymus leaf extract, Nephelium lapaceum leaf extract and excipients.

[0012] Furthermore, the excipients are water and polyols.

[0013] Furthermore, the polypeptide includes at least one of palmitoyl tripeptide-1, palmitoyl pentapeptide-4, palmitoyl tetrapeptide-7, tripeptide-1, acetyl hexapeptide-8, blue copper peptide, oligopeptide-1, oligopeptide-2, hexapeptide-3, and hexapeptide-9.

[0014] Furthermore, the polyol includes at least one of ethylene glycol, propylene glycol, glycerol, butylene glycol, hexanediol, and pentanediol.

[0015] Further, the composition, by mass percentage, comprises: 0.01-2% Sophora japonica pollen extract, 0.001-1% polypeptide, 0.01-1.5% artichoke leaf extract, 0.01-1.5% rambutan leaf extract, 15-40% polyol and the balance water.

[0016] Furthermore, the preparation method of artichoke leaf extract includes: pulverizing artichoke leaves, extracting with 50-70 wt% ethanol, filtering, concentrating, and freeze-drying. The above-mentioned 50-70 wt% ethanol extraction is beneficial in enhancing the effect of artichoke leaf extract on promoting collagen production.

[0017] Furthermore, the preparation method of rambutan leaf extract includes: pulverizing rambutan leaves, adding 35-60wt% ethanol for extraction, filtering, concentrating, and freeze-drying.

[0018] As mentioned above, the combination of artichoke leaf extract and rambutan leaf extract can produce a complementary effect, synergistically promoting the endogenous production of collagen, effectively increasing skin elasticity, and giving the skin a plump and youthful appearance. A more preferable weight ratio is 1:0.8-1.5 for artichoke leaf extract and rambutan leaf extract.

[0019] The present invention also provides a method for preparing the above composition, comprising heating a polyol, adding a polypeptide and stirring, then adding water and continuing to stir, adding sophora japonica pollen extract, artichoke leaf extract and rambutan leaf extract and stirring, sterilizing, and obtaining the composition.

[0020] The present invention also provides the use of the above composition in the preparation of cosmetics for firming and increasing skin elasticity. The composition has a weight concentration of 0.001-40% in the cosmetic.

[0021] The advantages of this invention compared to the prior art are: This invention provides a Sophora japonica flower powder extract for use in the preparation of compositions. The preparation method of the Sophora japonica flower powder extract includes: taking Sophora japonica flower powder, boiling it in water and then cooling it; adding a carbon source and Aspergillus oryzae CICC 2066 for fermentation; sterilization; filtration; concentration; and freeze-drying. It can effectively promote collagen production, increasing skin elasticity and firmness.

[0022] The Sophora japonica flower extract described in this invention can be used in food and cosmetics, and has good application prospects.

[0023] This invention uses Sophora japonica flower pollen extract to prepare a composition that effectively enhances the composition's effect on promoting collagen production. Furthermore, the fermentation method used to prepare the Sophora japonica flower pollen extract, compared to other extraction methods, further enhances the composition's collagen-promoting effect.

[0024] The present invention uses Aspergillus oryzae for fermentation, which can effectively improve the composition effect compared with fermentation by other bacteria. Furthermore, the use of a specific Aspergillus oryzae with the code CICC 2066 can further enhance the composition's ability to promote the synthesis of type I and type III collagen.

[0025] This invention uses Sophora japonica flower powder extract, polypeptides, artichoke leaf extract, rambutan leaf extract and excipients to prepare a composition that can effectively promote collagen production and increase skin elasticity and firmness.

[0026] This invention utilizes both artichoke leaf extract prepared with 50-70 wt% ethanol and rambutan leaf extract prepared with 35-60 wt% ethanol to effectively promote the endogenous production of type I and III collagen. The combination of these two extracts is more effective than using either one alone in promoting the production of type I and III collagen, as they work synergistically to enhance collagen production. The optimal ratio of 1:0.5-1.5 yields the best results, providing the skin with the firmest and plumpest texture.

[0027] The composition prepared by this invention can effectively promote collagen production at low concentrations, thereby enhancing skin elasticity and firmness. Detailed Implementation

[0028] To enable those skilled in the art to gain a clearer and more intuitive understanding of the present invention, the present invention will be further described below in conjunction with specific embodiments. Experimental methods in the following examples, unless otherwise specified, should be performed according to conventional methods and conditions, or according to the product instructions.

[0029] Example 1 Preparation method of artichoke (CYNARA SCOLYMUS) leaf extract: Artichoke leaves were crushed, and 60wt% ethanol was added at a weight ratio of 1:15. The mixture was stirred at 80rpm for 5 hours at 55℃, filtered through a 0.22μm filter membrane, concentrated under reduced pressure at 50℃ to 10.4% of the filtrate weight, and then freeze-dried at -50℃ to a water content of 1.42wt%.

[0030] Preparation method of rambutan (NEPHELIUM LAPPACEUM) leaf extract: Rambutan leaves were crushed, and 50wt% ethanol was added at a material-to-liquid ratio of 1:18. The mixture was stirred at 90rpm for 4 hours at 52℃, filtered through a 0.22μm filter membrane, concentrated under reduced pressure at 50℃ to 11.2% by weight of the filtrate, and then freeze-dried at -50℃ to a water content of 1.35wt%.

[0031] This study investigated the efficiency of the above-mentioned individual extracts and their combinations in promoting the production of type I and III collagen. The sample combinations are shown in Table 1. In Table 1, Q represents the weight ratio of artichoke leaf extract to rambutan leaf extract.

[0032] Table 1: Sample Composition The specific testing method is as follows: HSF cells in the logarithmic growth phase were taken, digested with trypsin, and a suspension was obtained. The suspension was then prepared at a ratio of 2 × 10⁶ cells per well. 5 Cells were seeded into 12-well plates, and culture medium was added to 1 mL. The plates were incubated at 37°C with 5% CO2 for 24 hours. The culture medium was then aspirated, and the plates were washed three times with PBS. The PBS wash buffer was then aspirated, and subsequent experiments were performed. For the control group, 150 μL of PBS was added to each well, followed by culture medium to 1 mL. For the experimental group, 150 μL of PBS was added to each well, followed by the sample (samples 1-11, with a final concentration of 100 ppm) and culture medium to 1 mL. All wells were incubated at 37°C with 5% CO2 for 72 hours. Each group was repeated four times. After incubation, the culture medium was aspirated, the plates were washed three times with PBS, and lysis buffer (RIPA to 1 mL) was added and shaken well. The plates were then centrifuged at 8000 rpm for 10 minutes. The supernatant was collected, and the content of type I and III collagen was tested according to the ELISA kit instructions. The results are shown in Table 2.

[0033] Table 2: Type I and Type III Collagen Content

[0034] Based on Table 2, calculate the increase in collagen content (Δ) in each group compared to the control group. The results are shown in Table 3.

[0035] Table 3: Δ values ​​of collagen in each group compared to the control group

[0036] Based on the test results in Tables 2-3, both the artichoke leaf extract and rambutan leaf extract prepared by the extraction method of this invention can effectively promote the endogenous production of type I and III collagen. A comparison of samples 1-2 and 3-11 shows that the combination of the two is more effective in promoting the production of type I and III collagen than using either one alone; they can synergistically enhance collagen production. Furthermore, data from the samples indicate that a ratio of 1:0.5-1.5 yields the best results, providing the optimal firming and plumping effect to the skin.

[0037] Example 2 The extract from Example 1 was used to prepare the composition and related tests were performed.

[0038] 1. Composition 1 The raw material composition, by weight percentage, is as follows: 1.5% Sophora japonica pollen extract, 0.08% polypeptide, 0.5% Artichoke leaf extract, 0.6% Rambutan leaf extract, 30% polyol, and the balance being water; the polypeptide is composed of palmitoyl tripeptide-1, palmitoyl pentapeptide-4, and palmitoyl tetrapeptide-7 in a weight ratio of 1:1:2; the polyol is composed of ethylene glycol, glycerol, and 1,3-butanediol in a weight ratio of 1:1:4.

[0039] The preparation method of Sophora japonica flower powder extract is as follows: Take Sophora japonica flower powder, add 20 times its weight of water, boil, and cool to room temperature. Add sucrose at 0.18% of the weight of the Sophora japonica flower powder and Aspergillus oryzae CICC 2066 (addition amount is 5×10). 7 Fermentation was carried out at 35℃ and 60 rpm for 4.5 days with sterile air introduced at a flow rate of 0.68 VVM. The mixture was then sterilized, filtered through a 0.22 μm pore size filter membrane, concentrated at 58℃ to 11.07% of the filtrate weight, and then freeze-dried at -52℃ to a water content of 1.95 wt%.

[0040] The composition is prepared as follows: a polyol is placed in a container and heated to 45°C. A polypeptide is added and stirred at 100 rpm for 10 minutes. Then water is added and stirring is continued at the same speed for 25 minutes. Sophora japonica pollen extract, artichoke leaf extract, and rambutan leaf extract are added and stirring is continued at the same speed for 15 minutes. The mixture is then sterilized to obtain the composition.

[0041] 2. Composition 2 The raw material composition, by weight percentage, is as follows: 1.3% Sophora japonica pollen extract, 0.095% polypeptide, 0.6% artichoke leaf extract, 0.62% rambutan leaf extract, 35% polyol, and the balance being water; the polypeptide consists of tripeptide-1, acetyl hexapeptide-8, and copper peptide in a weight ratio of 1:2:3; the polyol consists of ethylene glycol and 1,3-butanediol in a weight ratio of 1:3.

[0042] The preparation method of Sophora japonica flower powder extract is as follows: Take Sophora japonica flower powder, add 25 times its weight of water, boil, and cool to room temperature. Add 0.2% sucrose and Aspergillus oryzae CICC 2066 (8×10) based on the weight of the Sophora japonica flower powder. 7 Fermentation was carried out at 35℃ and 60 rpm for 3.5 days with sterile air introduced at a flow rate of 0.65 VVM. The mixture was then sterilized, filtered through a 0.22 μm pore size filter membrane, concentrated at 58℃ to 11.35% by weight of the filtrate, and then freeze-dried at -52℃ to a water content of 1.82 wt%.

[0043] The preparation method of composition 2 is the same as that of composition 1.

[0044] 3. Composition 3 The difference from Composition 1 is that it does not use Sophora japonica pollen extract: Raw material composition, by weight percentage: 0.08% polypeptide, 0.5% artichoke leaf extract, 0.6% rambutan leaf extract, 30% polyol and balance water; the polypeptide is composed of palmitoyl tripeptide-1, palmitoyl pentapeptide-4 and palmitoyl tetrapeptide-7 in a weight ratio of 1:1:2; the polyol is composed of ethylene glycol, glycerol and 1,3-butanediol in a weight ratio of 1:1:4.

[0045] The composition is prepared as follows: a polyol is placed in a container and heated to 45°C. A polypeptide is added and stirred at 100 rpm for 10 minutes. Then water is added and stirring is continued at the same speed for 25 minutes. Artichoke leaf extract and rambutan leaf extract are added and stirring is continued at the same speed for 15 minutes. The mixture is then sterilized to obtain the composition.

[0046] 4. Composition 4 The difference between this composition and composition 1 is that the Sophora japonica pollen extract is prepared by fermentation using a different Aspergillus oryzae number. Specifically, Aspergillus oryzae numbered GDMCC NO.3.31 is used to replace the Aspergillus oryzae in composition 1. All other aspects are the same.

[0047] 5. Composition 5 The difference between this composition and composition 1 is that the Sophora japonica pollen extract is prepared by fermentation using other bacteria. Specifically, Bacillus subtilis (GDMCC NO.1.258) is used to replace Aspergillus oryzae in composition 1. All other aspects are the same.

[0048] 6. Composition 6 The difference between this composition and composition 1 is that the Sophora japonica pollen extract is prepared by fermentation using other bacteria. Specifically, Acetobacter spp. with CICC 20441 replaces Aspergillus oryzae in composition 1. All other aspects are the same.

[0049] 7. Composition 7 The difference between this composition and composition 1 lies in the preparation method of the Sophora japonica pollen extract; all other aspects are the same.

[0050] The preparation method of Sophora japonica flower powder extract is as follows: Sophora japonica flower powder is taken, 15 times the mass of 25wt% ethanol is added, and the mixture is extracted at 45℃ with stirring at 80rpm for 4 hours. The mixture is filtered through a 0.22μm pore size filter membrane, concentrated at 54℃ to 11.35% by weight of filtrate, and then freeze-dried at -52℃ to a water content of 1.89wt%.

[0051] The above composition was subjected to performance testing to determine its efficiency in promoting collagen production.

[0052] HSF cells in the logarithmic growth phase were digested with trypsin to obtain a suspension, and then divided into two batches of 2 × 10⁶ cells per well. 5 Cells were seeded into 12-well plates, and culture medium was added to 1 mL. The plates were incubated at 37°C with 5% CO2 for 24 hours. The culture medium was then aspirated, and the cells were washed three times with PBS. The PBS wash buffer was then aspirated, and subsequent experiments were performed. For the control group, 150 μL of PBS was added to each well, followed by culture medium to 1 mL. For the experimental group, 150 μL of PBS was added to each well, followed by the combination (compositions 1-7, with a final total concentration of 0.45 wt%) and culture medium to 1 mL. All wells were incubated at 37°C with 5% CO2 for 72 hours. Each group was repeated four times. After incubation, the culture medium was aspirated, the cells were washed three times with PBS, and lysis buffer (RIPA to 1 mL) was added and shaken well. The plates were then centrifuged at 8000 rpm for 10 minutes. The supernatant was collected, and the content of type I and III collagen was measured according to the ELISA kit instructions. The results are shown in Table 4.

[0053] Table 4: Tests on the effect of promoting collagen production

[0054] According to the results of the 4-to-1 test, the composition prepared by the present invention can effectively promote collagen production at low concentrations, thereby enhancing skin elasticity and firmness.

[0055] A comparison of compositions 1, 3, and 7 shows that the composition prepared using Sophora japonica pollen extract in this invention can effectively enhance the composition's effect on promoting collagen production. Furthermore, the fermentation method used to prepare Sophora japonica pollen extract, compared to other extraction methods, further enhances the composition's effect on promoting collagen production.

[0056] Based on the comparison of compositions 1 and 4-6, it can be seen that the use of Aspergillus oryzae for fermentation in this invention can effectively improve the composition effect compared with fermentation by other bacteria. Furthermore, the use of Aspergillus oryzae with the specific number CICC 2066 can further enhance the composition's ability to promote the synthesis of type I and III collagen.

[0057] The above description of the embodiments is provided to enable those skilled in the art to understand and apply the present invention. It will be apparent to those skilled in the art that various modifications can be made to these embodiments, and the general principles described herein can be applied to other embodiments without inventive effort. Therefore, the present invention is not limited to the embodiments described herein, and any improvements and modifications made to the present invention by those skilled in the art based on simple variations or substitutions should be within the scope of protection of the present invention.

Claims

1. A sophora japonica pollen extract, characterized in that, The preparation method of Sophora japonica flower powder extract includes: taking Sophora japonica flower powder, adding water to boil and then cooling, adding carbon source and Aspergillus oryzae for fermentation, sterilization, filtration, concentration and freeze drying.

2. The sophora japonica pollen extract according to claim 1, characterized in that, Aspergillus oryzae is designated as CICC 2066.

3. The sophora japonica pollen extract according to claim 1, characterized in that, The amount of carbon source used is 0.01-3% of the weight of Sophora japonica flower powder, and the fermentation temperature and time are 25-40℃ for 3-5 days.

4. The sophora japonica pollen extract according to claim 1, characterized in that, The dosage of Aspergillus oryzae used for inoculation and fermentation is 10. 6 -10 11 One bacterium / g of Sophora japonica pollen.

5. A composition based on Sophora japonica pollen that promotes collagen production, characterized in that, The raw material composition is: Sophora japonica pollen extract, polypeptide, artichoke leaf extract, rambutan leaf extract and excipients as described in any one of claims 1-4; the excipients are water and polyol.

6. The composition according to claim 5, characterized in that, The polypeptide includes at least one of palmitoyl tripeptide-1, palmitoyl pentapeptide-4, palmitoyl tetrapeptide-7, tripeptide-1, acetyl hexapeptide-8, blue copper peptide, oligopeptide-1, oligopeptide-2, hexapeptide-3, and hexapeptide-9; And / or, the polyols include at least one of ethylene glycol, propylene glycol, glycerol, butylene glycol, hexanediol, and pentanediol.

7. The composition according to claim 5, characterized in that, The components, by weight percentage, are: 0.01-2% Sophora japonica pollen extract, 0.001-1% polypeptide, 0.01-1.5% artichoke leaf extract, 0.01-1.5% rambutan leaf extract, 15-40% polyol, and the balance being water.

8. The composition according to claim 7, characterized in that, The weight ratio of artichoke leaf extract to rambutan leaf extract is 1:0.8-1.5; And / or, the preparation method of artichoke leaf extract includes: crushing artichoke leaves, adding 50-70wt% ethanol for extraction, filtering, concentrating, and freeze-drying to obtain the extract; And / or, the preparation method of rambutan leaf extract includes: pulverizing rambutan leaves, adding ethanol with a concentration of 35-60wt% for extraction, filtering, concentrating, and freeze-drying.

9. A method for preparing the composition according to any one of claims 5-8, characterized in that, The process involves heating a polyol, adding a polypeptide and stirring, then adding water and continuing to stir, adding sophora japonica pollen extract, artichoke leaf extract, and rambutan leaf extract and stirring, followed by sterilization to obtain the composition.

10. The use of the composition according to any one of claims 5-8 in the preparation of a product that promotes collagen production.

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