A composition and mixture for promoting endogenous collagen production and applications

Abstract

In order to solve the problems of the prior art, the application provides a composition for promoting endogenous collagen production, which comprises a main component and an auxiliary component, the auxiliary component is added in an amount of 5-7% of the mass of the main component, wherein the main component comprises silk fibroin and peptides; and the auxiliary component is Panax notoginseng extract.The peptides comprise at least one of carnosine, tetrapeptide, acetyl hexapeptide and arginine / lysine polypeptide.The application significantly promotes the production of type I collagen in cells, and through tests, the content of type I collagen contained in human dermal fibroblasts in the experimental group using the composition for promoting endogenous collagen production of the application is increased by more than 70% compared with the blank group in an experimental cycle.

Description

Technical Field

[0001] This invention relates to the field of daily chemical products technology, and in particular to a composition, mixture, and application that promotes the production of endogenous collagen. Background Technology

[0002] Collagen is the most abundant protein in the human body, found in bones, muscles, skin, and tendons, and is the substance that holds the body together. As we age, the rate of collagen loss accelerates. If collagen is not replenished in time, the skin will lose its elasticity, resulting in wrinkles and sagging. Therefore, how to more effectively replenish collagen in the skin has always been an important research direction in cosmetics.

[0003] There are two main ways to supplement collagen in the skin: endogenous and exogenous. Endogenous collagen supplementation mainly works by promoting the body's own collagen production, which is more compatible with the body and has almost no rejection reaction.

[0004] Current methods for supplementing endogenous collagen in the skin primarily involve selective dietary intake or direct copper supplementation to promote the body's own collagen production. However, existing technologies are relatively slow to take effect and require long-term adherence to a regulated diet or the use of copper ion supplements, resulting in low efficiency and inconvenience. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention provides a composition for promoting endogenous collagen production, comprising: a main component and an auxiliary component, wherein the amount of the auxiliary component added is 5-7% of the mass of the main component, wherein: the main component includes: silk fibroin and peptides; and the auxiliary component is: Panax notoginseng extract.

[0006] Furthermore, the peptide includes at least one of carnosine, tetrapeptide, acetyl hexapeptide, and arginine / lysine polypeptide.

[0007] Furthermore, by mass parts, the peptides comprise: 15-20 parts carnosine, 8-12 parts tetrapeptide-3, 10-15 parts acetyl hexapeptide-8, and 10-20 parts arginine / lysine polypeptide.

[0008] Furthermore, by mass, the main components include: 100 parts silk fibroin and 10-50 parts peptides.

[0009] Preferably, the main components, by mass parts, include: 100 parts silk fibroin and 18.75-42.85 parts peptides.

[0010] Furthermore, the Panax notoginseng extract is a substance obtained by extracting fresh Panax notoginseng using deionized water and / or ethanol as solvents.

[0011] Furthermore, the preparation method of the Panax notoginseng extract includes: (1) Place the crushed Panax notoginseng into an extraction tank and add 7-10 times the weight of the first extract of Panax notoginseng; (2) In an environment of 30-40℃, the liquid phase components were circulated for 5-7 hours to obtain the first extraction product liquid; (3) Add 10-12 times the weight of the second extract of Panax notoginseng and circulate the extract at 95-100℃ for 2-4 hours; drain the liquid phase components and cool to obtain the second extract product liquid; (4) The first extraction product liquid and the second extraction product liquid are mixed to obtain an extraction mixture; (5) The extract mixture is freeze-dried to obtain the Panax notoginseng extract.

[0012] Furthermore, by mass parts, the first extract comprises: 30-35 parts deionized water and 100 parts ethanol; the second extract is deionized water.

[0013] Furthermore, during step (2), an 800-1000MHz microwave field and a 345-400MHz microwave field are alternately applied to the outside of the first extract. The duration of the 800-1000MHz microwave field is 2-3 minutes, and the duration of the 345-400MHz microwave field is 3-5 minutes. One cycle consists of applying the 800-1000MHz microwave field and one cycle of applying the 345-400MHz microwave field, and this cycle is repeated 4-5 times to complete the extraction operation in step (2).

[0014] Furthermore, before freeze-drying the extract mixture, the process includes: first, placing the extract mixture into a stirring tank and continuously stirring at a low speed; then, using a vacuum pump to continuously extract the gas in the stirring tank until the liquid level in the stirring tank drops to 60-65% of the original liquid level; after restoring the gas pressure in the stirring tank to atmospheric pressure, the remaining liquid in the stirring tank is concentrated through a membrane concentration device to obtain the extract concentrate; the extract concentrate is used for freeze-drying powdering in step (5).

[0015] Furthermore, by weight percentage, the auxiliary ingredients include: 63-67% Panax notoginseng extract and the balance being saffron extract.

[0016] Furthermore, the saffron extract is obtained by ethanol extraction of dried saffron.

[0017] Furthermore, the preparation method of the saffron extract includes: (a) After pretreatment, saffron is placed in an extraction tank equipped with a crusher, and a third extract of 3-4 times the weight of saffron is added; the third extract comprises, by mass percentage, 50-55% water and the remainder ethanol; (ii) In an environment of 50-60℃, circulate and extract for 8-10 hours, and then discharge the liquid phase components to obtain the third extraction product liquid; (iii) The third extract product liquid is freeze-dried and powdered to obtain the saffron extract.

[0018] Furthermore, the pretreatment is as follows: the dried saffron is placed in an acidic aqueous solution, so that the dried saffron is completely submerged in the acidic aqueous solution, and after soaking for 20-30 hours, it is washed with clean water to complete the pretreatment; the acidic aqueous solution is an acidic aqueous solution with a pH of 4.0-5.0.

[0019] Preferably, the acidic aqueous solution is an acetic acid aqueous solution.

[0020] Furthermore, during step (ii), a microwave field of 575-615MHz is continuously applied, reducing the extraction time to 30-40min.

[0021] The second objective of this invention is to provide a mixture that promotes the production of endogenous collagen. The mixture is prepared by mixing 0.01-2‰ of the composition with the remainder of deionized water by weight; the composition is the same as the composition described above for promoting the production of endogenous collagen.

[0022] A third objective of this invention is to provide an application of a composition that promotes the production of endogenous collagen, specifically: applying the above-mentioned composition that promotes the production of endogenous collagen to skin care products.

[0023] The advantages of this invention are: This invention significantly promotes the production of type I collagen in cells. After testing, within one experimental cycle, the experimental group using the composition of this invention that promotes the production of endogenous collagen showed that the content of type I collagen in human dermal fibroblasts was increased by more than 70% compared with the blank group, which is a significant effect. Detailed Implementation

[0024] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. 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.

[0025] Example 1 A composition for promoting endogenous collagen production includes: a main component and an auxiliary component, wherein the auxiliary component is added at 6% of the mass of the main component, wherein: the main component includes: silk fibroin and peptides; and the auxiliary component is: Panax notoginseng extract.

[0026] By mass fractions, the peptides comprise: 18 parts carnosine, 10 parts tetrapeptide-3, 12 parts acetyl hexapeptide-8, and 15 parts arginine / lysine polypeptide.

[0027] By weight, the main components include: 100 parts silk fibroin and 30 parts peptide.

[0028] The Panax notoginseng extract is a substance obtained by extracting fresh Panax notoginseng using deionized water and / or ethanol as solvents.

[0029] Example 2 A composition for promoting endogenous collagen production includes: a main component and an auxiliary component, wherein the auxiliary component is added at 7% of the mass of the main component, wherein: the main component includes: silk fibroin and peptides; and the auxiliary component is: Panax notoginseng extract.

[0030] By mass fractions, the peptides comprise: 20 parts carnosine, 12 parts tetrapeptide-3, 15 parts acetyl hexapeptide-8, and 20 parts arginine / lysine polypeptide.

[0031] The main components, by mass, include: 100 parts silk fibroin and 50 parts peptide.

[0032] The Panax notoginseng extract is a substance obtained by extracting fresh Panax notoginseng using deionized water and / or ethanol as solvents.

[0033] Example 3 A composition for promoting endogenous collagen production includes: a main component and an auxiliary component, wherein the auxiliary component is added at 5% of the mass of the main component, wherein: the main component includes: silk fibroin and peptides; and the auxiliary component is: Panax notoginseng extract.

[0034] By mass fractions, the peptides comprise: 15 parts carnosine, 8 parts tetrapeptide-3, 10 parts acetyl hexapeptide-8, and 10 parts arginine / lysine polypeptide.

[0035] By weight, the main components include: 100 parts silk fibroin and 10 parts peptide.

[0036] The Panax notoginseng extract is a substance obtained by extracting fresh Panax notoginseng using deionized water and / or ethanol as solvents.

[0037] Comparative Example 1-1 A composition for promoting endogenous collagen production includes: a main component and an auxiliary component, wherein the auxiliary component is added at 6% of the mass of the main component, wherein: the main component includes: silk fibroin and peptides; and the auxiliary component is: Panax notoginseng extract.

[0038] The peptide comprises, by mass parts, 55 parts carnosine.

[0039] By weight, the main components include: 100 parts silk fibroin and 30 parts peptide.

[0040] The Panax notoginseng extract is a substance obtained by extracting fresh Panax notoginseng using deionized water and / or ethanol as solvents.

[0041] Comparative Examples 1-2 A composition for promoting endogenous collagen production includes: 100 parts silk fibroin and 30 parts peptides.

[0042] The peptide comprises, by mass parts, 55 parts carnosine.

[0043] Comparative Example 2-1 A composition for promoting endogenous collagen production includes: a main component and an auxiliary component, wherein the auxiliary component is added at 6% of the mass of the main component, wherein: the main component includes: silk fibroin and peptides; and the auxiliary component is: Panax notoginseng extract.

[0044] The peptide comprises, by mass parts, 55 parts tetrapeptide-3.

[0045] By weight, the main components include: 100 parts silk fibroin and 30 parts peptide.

[0046] The Panax notoginseng extract is a substance obtained by extracting fresh Panax notoginseng using deionized water and / or ethanol as solvents.

[0047] Comparative Example 2-2 A composition for promoting endogenous collagen production includes: 100 parts silk fibroin and 30 parts peptides.

[0048] The peptide comprises, by mass parts, 55 parts tetrapeptide-3.

[0049] Comparative Example 3-1 A composition for promoting endogenous collagen production includes: a main component and an auxiliary component, wherein the auxiliary component is added at 6% of the mass of the main component, wherein: the main component includes: silk fibroin and peptides; and the auxiliary component is: Panax notoginseng extract.

[0050] The peptide comprises, by mass parts, 55 parts of acetyl hexapeptide-8.

[0051] By weight, the main components include: 100 parts silk fibroin and 30 parts peptide.

[0052] The Panax notoginseng extract is a substance obtained by extracting fresh Panax notoginseng using deionized water and / or ethanol as solvents.

[0053] Comparative Example 3-2 A composition for promoting endogenous collagen production includes: 100 parts silk fibroin and 30 parts peptides.

[0054] The peptide comprises, by mass parts, 55 parts of acetyl hexapeptide-8.

[0055] Comparative Example 4-1 A composition for promoting endogenous collagen production includes: a main component and an auxiliary component, wherein the auxiliary component is added at 6% of the mass of the main component, wherein: the main component includes: silk fibroin and peptides; and the auxiliary component is: Panax notoginseng extract.

[0056] The peptide comprises, by mass parts, 55 parts of arginine / lysine polypeptide.

[0057] By weight, the main components include: 100 parts silk fibroin and 30 parts peptide.

[0058] The Panax notoginseng extract is a substance obtained by extracting fresh Panax notoginseng using deionized water and / or ethanol as solvents.

[0059] Comparative Example 4-2 A composition for promoting endogenous collagen production includes: 100 parts silk fibroin and 30 parts peptides.

[0060] The peptide comprises, by mass parts, 55 parts of arginine / lysine polypeptide.

[0061] Comparative Example 5-1 A composition for promoting endogenous collagen production includes: a main component and an auxiliary component, wherein the auxiliary component is added at 6% of the mass of the main component, wherein: the main component includes: silk fibroin and peptides; and the auxiliary component is: Panax notoginseng extract.

[0062] The peptides comprise, by mass parts, 16 parts tetrapeptide-3, 18 parts acetyl hexapeptide-8 and 21 parts arginine / lysine polypeptide.

[0063] By weight, the main components include: 100 parts silk fibroin and 30 parts peptide.

[0064] The Panax notoginseng extract is a substance obtained by extracting fresh Panax notoginseng using deionized water and / or ethanol as solvents.

[0065] Comparative Example 5-2 A composition for promoting endogenous collagen production includes: 100 parts silk fibroin and 30 parts peptides.

[0066] The peptides comprise, by mass parts, 16 parts tetrapeptide-3, 18 parts acetyl hexapeptide-8 and 21 parts arginine / lysine polypeptide.

[0067] Comparative Example 6-1 A composition for promoting endogenous collagen production includes: a main component and an auxiliary component, wherein the auxiliary component is added at 6% of the mass of the main component, wherein: the main component includes: silk fibroin and peptides; and the auxiliary component is: Panax notoginseng extract.

[0068] The peptides comprise, by mass parts, 21 parts carnosine, 15 parts acetyl hexapeptide-8 and 19 parts arginine / lysine polypeptide.

[0069] By weight, the main components include: 100 parts silk fibroin and 30 parts peptide.

[0070] The Panax notoginseng extract is a substance obtained by extracting fresh Panax notoginseng using deionized water and / or ethanol as solvents.

[0071] Comparative Example 6-2 A composition for promoting endogenous collagen production includes: 100 parts silk fibroin and 30 parts peptides.

[0072] The peptides comprise, by mass parts, 21 parts carnosine, 15 parts acetyl hexapeptide-8 and 19 parts arginine / lysine polypeptide.

[0073] Comparative Example 7-1 A composition for promoting endogenous collagen production includes: a main component and an auxiliary component, wherein the auxiliary component is added at 6% of the mass of the main component, wherein: the main component includes: silk fibroin and peptides; and the auxiliary component is: Panax notoginseng extract.

[0074] The peptide comprises, by mass parts, 25 parts acetyl hexapeptide-8 and 30 parts arginine / lysine polypeptide.

[0075] By weight, the main components include: 100 parts silk fibroin and 30 parts peptide.

[0076] The Panax notoginseng extract is a substance obtained by extracting fresh Panax notoginseng using deionized water and / or ethanol as solvents.

[0077] Comparative Example 7-2 A composition for promoting endogenous collagen production includes: 100 parts silk fibroin and 30 parts peptides.

[0078] The peptide comprises, by mass parts: 25 parts acetyl hexapeptide-8 and 30 parts arginine / lysine polypeptide.

[0079] Comparative Example 8 A composition for promoting endogenous collagen production includes: 100 parts silk fibroin and 30 parts peptides.

[0080] By mass fractions, the peptides comprise: 18 parts carnosine, 10 parts tetrapeptide-3, 12 parts acetyl hexapeptide-8, and 15 parts arginine / lysine polypeptide.

[0081] The compositions obtained in Example 1 and Comparative Examples 1-7 are labeled as Composition A (Example 1), Composition B1-1 (Comparative Example 1-1), Composition B1-2 (Comparative Example 1-2), Composition B2-1 (Comparative Example 2-1), Composition B2-2 (Comparative Example 2-2), Composition B3-1 (Comparative Example 3-1), Composition B3-2 (Comparative Example 3-2), Composition B4-1 (Comparative Example 4-1), Composition B4-2 (Comparative Example 4-2), Composition B5-1 (Comparative Example 5-1), Composition B5-2 (Comparative Example 5-2), Composition B6-1 (Comparative Example 6-1), Composition B6-2 (Comparative Example 6-2), Composition B7-1 (Comparative Example 7-1), Composition B7-2 (Comparative Example 7-2), and Composition B8 (Comparative Example 8), respectively. A type I collagen experimental group was constructed using the above compositions, specifically including: Control group: Human dermal fibroblasts (HSF) were selected as the cell model, and 1.8 × 10⁻⁶ cells were used. 5 Cells were seeded at a density of 1 cell per well in 6-well plates and incubated overnight at 37°C with 5% CO2. When the cell deposition rate in the 6-well plates reached 40%-60%, 2 mL of DMEM complete medium was added to each well to obtain the control group.

[0082] Experimental group: Human dermal fibroblasts (HSF) were selected as the cell model, and 1.8 × 10⁻⁶ cells were used. 5 Cells were seeded at a density of 1 cell per well in 6-well plates and incubated overnight at 37°C with 5% CO2. When the cell deposition rate in the 6-well plates reached 40%-60%, 2 mL of DMEM complete medium containing composition A (concentration of composition A was 0.01‰) was added to each well to obtain experimental group A.

[0083] Human dermal fibroblasts (HSF) were selected as the cell model, and 1.8 × 10⁻⁶ cells were used. 5 Cells were seeded at a density of 1 cell per well in 6-well plates and incubated overnight at 37°C with 5% CO2. When the cell deposition rate in the 6-well plates reached 40%-60%, 2 mL of DMEM complete medium containing composition B1-1 (concentration of composition B1-1 was 0.1‰) was added to each well to obtain experimental group B1-1.

[0084] The same method was used to obtain experimental groups B1-2, B2-1, B2-2, B3-1, B3-2, B4-1, B4-2, B5-1, B5-2, B6-1, B6-2, B7-1, B7-2, and B8.

[0085] After the culture medium was added to the control group and the sample group, all the 6-well plates were placed in an incubator and cultured at 37°C and 5% CO2 for 48 h. The culture medium was then aspirated, the cells were washed twice with PBS, digested with 0.25% trypsin, centrifuged at 3000 rpm for 5 min, and the supernatant was collected in EP tubes. The cells were washed once with PBS, centrifuged again, and the supernatant was collected in EP tubes to obtain the sample to be tested.

[0086] The ELISA kit was used to test the content of type I collagen in the sample to be tested.

[0087] The results of type I collagen content testing in the control group and experimental groups B1-1 to B8 are shown in the table below: Table 1. Results of Type I Collagen Content Test in Control Group and Experimental Groups B1-1 to B8 Type I collagen growth rate (%) = (Mn - M0) / M0 100%, where Mn is the type I collagen content in the experimental group and M0 is the type I collagen content in the control group.

[0088] The results of the type I collagen content test show that: 1. Pairwise comparisons of the test results from experimental groups A and B8, experimental groups B1-1 and B1-2, experimental groups B2-1 and B2-2, ..., experimental groups B6-1 and B6-2 show that the addition of the adjuvant Panax notoginseng has a very significant synergistic effect on the experimental groups containing tetrapeptide-3. For example, after adding Panax notoginseng, the type I collagen content between experimental groups A and B8 increased from 75.34 ng / mL in experimental group B8 to... The level of type I collagen in experimental groups A was 94.58 ng / mL. Between experimental groups B2-1 and B2-2, the addition of Panax notoginseng increased the type I collagen content from 65.37 ng / mL in group B2-2 to 79.52 ng / mL in group B2-1. Similarly, between experimental groups B5-1 and B5-2, the addition of Panax notoginseng increased the type I collagen content from 70.12 ng / mL in group B5-2 to 86.27 ng / mL in group B5-1, demonstrating a very significant effect. In the other experimental groups without tetrapeptide-3, the difference before and after adding Panax notoginseng was not significant.

[0089] 2. The comparison between the control group and the experimental group B1-2 shows that carnosine itself has little effect on the content of type I collagen. The main reason is that when carnosine is used alone, it mainly acts as a free radical scavenger. It mainly prevents the free radicals from binding with type I collagen and thus reducing the activity of type I collagen by capturing free radicals.

[0090] 3. Comparison of experimental groups B1-2, B2-2, B3-2, B4-2, B5-2, B6-2, B7-2, and B8 shows that the effects of tetrapeptide-3, acetyl hexapeptide-8, and arginine / lysine peptides, used alone or in combination, on the growth rate of type I collagen were relatively similar, without a significant increase. When carnosine was used in combination with acetyl hexapeptide-8 and arginine / lysine peptides (experimental group B6-2), its effect was weaker than that of the same composition of acetyl hexapeptide-8 and arginine / lysine peptides (experimental group B7-2). This is because carnosine's effect on the growth rate of type I collagen is weaker than that of acetyl hexapeptide-8 or arginine / lysine peptides alone. However, when carnosine was used in combination with tetrapeptide-3, acetyl hexapeptide-8, and arginine / lysine peptides, the growth rate of type I collagen actually increased, from 70.12 ng / mL in experimental group B5-2 to 75.34 ng / mL in experimental group B8. This may be due to a synergistic effect between carnosine and the combination of tetrapeptide-3, acetyl hexapeptide-8, and arginine / lysine peptides. Therefore, even though carnosine occupied a portion of the composition, the deteriorating effect of experimental group B6-2 was transformed into an optimizing effect in experimental group B8.

[0091] Furthermore, based on Example 1, the applicant used a single-factor experimental method, keeping the silk fibroin content of the main component constant at 100 parts, and proportionally changing the total amount of peptides added to: 0, 10, 20, 30, 40, 50, 60, 70, 80, 100, 200, and 300 parts. The results of the type I collagen content test are shown in the table below: Table 2. Test results of type I collagen content in products with different peptide contents. As shown in the table above, when the compound peptides of this invention are added to silk fibroin, the effect of the product of this invention on increasing the content of type I collagen can be significantly improved even at a low addition amount. When the addition amount exceeds 50 parts, the improvement effect declines and gradually tends to stabilize. It can be seen that only within a specific addition range can the product of this invention achieve a significant effect on increasing the content of type I collagen. Adding too much not only increases the cost, but also causes the effect to gradually decrease and then tend to stabilize.

[0092] Based on the above experiments, the applicant further determined that when the amount of silk fibroin added is 18.75-42.85 parts, the product of the present invention has the most significant effect on increasing the content of type I collagen, especially when the amount added is 30 parts, the effect is the best.

[0093] Regarding the amount of auxiliary ingredients added, the applicant adjusted the amount of Panax notoginseng extract added based on Example 1 and found that when the amount of Panax notoginseng extract added was less than 5% (less than or equal to 4%), the detection value of type I collagen content showed a significant decrease. The specific test results are shown in the table below: Table 3. Test results of type I collagen content in products with different amounts of Panax notoginseng extract. As shown in the table above, the amount of auxiliary ingredients added needs to be strictly controlled. The relationship between the amount added and the final effect is non-linear. The best effect is achieved when the amount of auxiliary ingredients added is 5-7%. When the amount added is too low, although there is still an improvement compared to the control group, the improvement is relatively small. When the amount added is too high, a deterioration phenomenon occurs.

[0094] Example 4 Based on any of the compositions in Examples 1-3 that promote endogenous collagen production, the method for preparing the Panax notoginseng extract includes: (1) Place the crushed Panax notoginseng into an extraction tank and add the first extract liquid, which is 8 times the weight of Panax notoginseng; (2) In an environment of 35℃, the liquid phase components were discharged to obtain the first extraction product liquid; (3) Add 11 times the mass of the second extract of Panax notoginseng and circulate the extract at 96°C for 3 hours; drain the liquid phase components and cool to obtain the second extract product liquid; (4) The first extraction product liquid and the second extraction product liquid are mixed to obtain an extraction mixture; (5) The extract mixture is freeze-dried to obtain the Panax notoginseng extract.

[0095] By mass fractions, the first extract comprises: 32 parts deionized water and 100 parts ethanol; the second extract is deionized water.

[0096] Example 5 Based on any of the compositions in Examples 1-3 that promote endogenous collagen production, the method for preparing the Panax notoginseng extract includes: (1) Place the crushed Panax notoginseng into an extraction tank and add the first extract liquid, which is 10 times the weight of Panax notoginseng; (2) In an environment of 40℃, the liquid phase components were discharged to obtain the first extraction product liquid; (3) Add 12 times the mass of the second extract of Panax notoginseng and circulate the extract at 100°C for 4 hours; drain the liquid phase components and cool to obtain the second extract product liquid; (4) The first extraction product liquid and the second extraction product liquid are mixed to obtain an extraction mixture; (5) The extract mixture is freeze-dried to obtain the Panax notoginseng extract.

[0097] By mass fractions, the first extract comprises: 35 parts deionized water and 100 parts ethanol; the second extract is deionized water.

[0098] Example 6 Based on any of the compositions in Examples 1-3 that promote endogenous collagen production, the method for preparing the Panax notoginseng extract includes: (1) Place the crushed Panax notoginseng into an extraction tank and add the first extract solution, which is 7 times the weight of Panax notoginseng; (2) In an environment of 30°C, the liquid phase components were circulated and extracted for 5 hours to obtain the first extraction product liquid; (3) Add 10 times the mass of the second extract of Panax notoginseng and circulate the extract at 95°C for 2 hours; drain the liquid phase components and cool to obtain the second extract product liquid; (4) The first extraction product liquid and the second extraction product liquid are mixed to obtain an extraction mixture; (5) The extract mixture is freeze-dried to obtain the Panax notoginseng extract.

[0099] By mass fractions, the first extract comprises: 30 parts deionized water and 100 parts ethanol; the second extract is deionized water.

[0100] Comparative Example 9 Based on any of the compositions in Examples 1-3 that promote endogenous collagen production, the method for preparing the Panax notoginseng extract includes: (1) Place the crushed Panax notoginseng into an extraction tank and add the first extract liquid, which is 11 times the weight of Panax notoginseng; (2) The mixture was extracted by circulating it at 35°C for 9 hours to obtain the extract mixture; (3) The extract mixture is freeze-dried to obtain the Panax notoginseng extract.

[0101] The first extract comprises, by mass parts, 32 parts of deionized water and 100 parts of ethanol.

[0102] Comparative Example 10 Based on any of the compositions in Examples 1-3 that promote endogenous collagen production, the method for preparing the Panax notoginseng extract includes: (1) Place the crushed Panax notoginseng into an extraction tank and add a second extract solution with a mass of 11 times that of Panax notoginseng; (2) The mixture was extracted by circulating it at 96°C for 9 hours to obtain the extract mixture; (3) The extract mixture is freeze-dried to obtain the Panax notoginseng extract.

[0103] The second extract is deionized water by mass fraction.

[0104] The Panax notoginseng extract described in Example 1 was obtained using the method described in Example 4, and was designated as experimental group C. The Panax notoginseng extracts obtained using the methods described in Comparative Examples 9 and 10 were used as auxiliary components. The products prepared using the proportions described in Example 1 were designated as control groups C-1 and C-2. The results of the type I collagen content test are as follows: Table 4. Results of Type I Collagen Content Test in Experimental Group C, Control Group C-1, and Control Group C-2 As shown in the table above, the synergistic effect of the Panax notoginseng extract obtained by using only a mixture of deionized water and ethanol, and the Panax notoginseng extract obtained entirely from deionized water, on tetrapeptide-3 was significantly weakened. In particular, the Panax notoginseng extract obtained from deionized water only showed a slightly higher effect than group B8, which did not contain any Panax notoginseng extract. Therefore, only the Panax notoginseng extract obtained using the extraction method of this invention can achieve a significant synergistic effect with tetrapeptide-3.

[0105] Based on Example 4, the applicant kept the amount of ethanol added to the first extract constant at 100 parts, and changed the amount of deionized water added to 0, 10, 20, 30, 40, 50, 60, 70, 80, 100, 200, and 300 parts. The Panax notoginseng extracts obtained from the first extracts with different ratios were used as experimental groups D-1, D-2, D-3, D-4, D-5, D-6, D-7, D-8, D-9, D-10, D-11, D-12, D-13, and D-14. The results of the type I collagen content test are as follows: Table 5. Results of Type I Collagen Content Test in Experimental Groups D-1 to D-18 As shown in the table above, with a constant ethanol content of 100 parts and the addition of 30-40 parts deionized water, the product exhibits a significant effect. With continued addition of deionized water, the product's effect gradually approaches that of pure deionized water. This demonstrates that only when the first extract is formulated in a specific ratio can a Panax notoginseng extract exhibiting good synergistic effects with tetrapeptide-3 be obtained.

[0106] The applicant confirmed through further experiments that the effect was most significant when the amount of ethanol was constant at 100 parts and the amount of deionized water added was 30-35 parts. In particular, when the amount of deionized water added was 32 parts, the content of type I collagen could reach 94.58 ng / mL.

[0107] Example 7 Based on the composition for promoting endogenous collagen production in Example 4, during step (2), a 927 MHz microwave field and a 382 MHz microwave field were alternately applied to the outside of the first extract. The duration of the 927 MHz microwave field was 3 minutes, and the duration of the 382 MHz microwave field was 4 minutes. One cycle of applying the 927 MHz microwave field and one cycle of applying the 382 MHz microwave field constituted one round, and this process was repeated four times to complete the extraction operation in step (2).

[0108] Example 8 Based on the composition for promoting endogenous collagen production in Example 4, during step (2), a 1000 MHz microwave field and a 400 MHz microwave field were alternately applied to the outside of the first extract. The duration of the 1000 MHz microwave field was 3 minutes, and the duration of the 400 MHz microwave field was 5 minutes. One cycle consisted of applying the 1000 MHz microwave field and one cycle of applying the 400 MHz microwave field, and this process was repeated 4-5 times to complete the extraction operation in step (2).

[0109] Example 9 Based on the composition for promoting endogenous collagen production in Example 4, during step (2), an 800 MHz microwave field and a 345 MHz microwave field were alternately applied to the outside of the first extract. The duration of the 800 MHz microwave field was 2 minutes, and the duration of the 345 MHz microwave field was 3 minutes. One cycle consisted of applying the 800 MHz microwave field and one cycle of applying the 345 MHz microwave field, and this process was repeated four times to complete the extraction operation in step (2).

[0110] Comparative Example 11 Based on the composition of Example 4 that promotes endogenous collagen production, when performing step (2), a microwave field of 927 MHz is applied to the outside of the first extract for a duration of 28 min.

[0111] Comparative Example 12 Based on the composition of Example 4 that promotes endogenous collagen production, when performing step (2), a microwave field of 382 MHz is applied to the outside of the first extract for a duration of 28 min.

[0112] Using Example 7 as experimental group D, Comparative Example 11 as experimental group D-1, and Comparative Example 12 as experimental group D-2, the type I collagen content was tested. The results are shown in the table below: Table 6. Results of Type I Collagen Content Test in Experimental Groups C, D, D-1, and D-2 According to general principles in the art, applying an external microwave field can significantly improve extraction efficiency. However, in this application, the applicant has accidentally discovered that, compared to the traditional constant-power microwave extraction method, the present invention, by regularly changing the frequency of the microwave field during the extraction process, can further enhance the synergistic effect between Panax notoginseng extract and the peptide composition of the present invention within the same extraction time.

[0113] Example 10 Based on any of the compositions for promoting endogenous collagen production in Examples 7-9, the extraction mixture is further prepared before freeze-drying by: firstly, placing the extraction mixture into a stirring tank and continuously stirring at a low speed; then continuously extracting the gas in the stirring tank using a vacuum pump until the liquid level in the stirring tank drops to 60-65% of the original liquid level; after restoring the gas pressure in the stirring tank to atmospheric pressure, concentrating the remaining liquid in the stirring tank through a membrane concentration device to obtain the extraction concentrate; the extraction concentrate is used for freeze-drying powdering in step (5).

[0114] This step, through pre-concentration, can significantly reduce the moisture content of freeze-dried products and freeze-drying time, thereby improving overall production efficiency.

[0115] Example 11 Based on the composition for promoting endogenous collagen production in Example 4, the auxiliary components, by weight percentage, include: 65% Panax notoginseng extract and the balance being saffron extract.

[0116] The preparation method of the saffron extract includes: (a) After pretreatment, saffron is placed in an extraction tank equipped with a crusher, and a third extract of 4 times the weight of saffron is added; the third extract comprises, by mass percentage, 53% water and the remainder ethanol; (ii) The third extract was obtained by circulating the extract at 55°C for 9 hours and then removing the liquid phase components. (iii) The third extract product liquid is freeze-dried and powdered to obtain the saffron extract.

[0117] Example 12 Based on the composition for promoting endogenous collagen production in Example 4, the auxiliary components, by weight percentage, include: 67% Panax notoginseng extract and the balance being saffron extract.

[0118] The preparation method of the saffron extract includes: (a) After pretreatment, saffron is placed in an extraction tank equipped with a crusher, and a third extract of 4 times the weight of saffron is added; the third extract comprises, by mass percentage, 55% water and the remainder ethanol; (ii) In an environment of 60℃, the mixture was circulated for 10 hours, and the liquid phase components were discharged to obtain the third extraction product liquid; (iii) The third extract product liquid is freeze-dried and powdered to obtain the saffron extract.

[0119] Example 13 Based on the composition for promoting endogenous collagen production in Example 4, the auxiliary components, by weight percentage, include: 63% Panax notoginseng extract and the balance being saffron extract.

[0120] The preparation method of the saffron extract includes: (a) After pretreatment, saffron is placed in an extraction tank equipped with a crusher, and a third extract solution with a mass ratio of 3 times that of saffron is added; the third extract solution comprises 50% water and the remainder ethanol by mass percentage. (ii) In an environment of 50℃, the mixture was circulated for 8 hours, and the liquid phase components were discharged to obtain the third extraction product liquid; (iii) The third extract product liquid is freeze-dried and powdered to obtain the saffron extract.

[0121] Comparative Example 13 The saffron extract extraction method of Example 12 was used, and the auxiliary component in the product was pure saffron extract of the same mass as the compound of Panax notoginseng extract and saffron extract in Example 12.

[0122] Using Example 11 as experimental group E and Comparative Example 13 as experimental group E-1, the results of the type I collagen content test are as follows: Table 7. Results of Type I Collagen Content Test in Experimental Groups C, E, and E-1 As can be seen from the table above, saffron extract alone does not have a significant synergistic effect on the silk fibroin and complex peptide composition of the present invention. However, when saffron extract and Panax notoginseng extract are used together, the efficacy of the product composition is further enhanced.

[0123] The applicant accidentally discovered that the combined use of saffron extract and Panax notoginseng extract had a better effect on promoting collagen production in the composition of this invention compared to using Panax notoginseng extract alone. The applicant conducted a comparative experiment (Comparative Example 13) and found that saffron alone did not produce a promoting effect and needed to be used in combination with Panax notoginseng extract. Therefore, the applicant conducted single-factor experiments and found that the effect was better when the amount of saffron extract added was 30-40% of the total amount of auxiliary ingredients, especially at 33-37%.

[0124] Furthermore, regarding the extract of saffron extract, the applicant attempted to use a segmented extraction method similar to that used for Panax notoginseng extract. They found that for saffron extract, the effects of using an ethanol + water extract followed by a water extract, an existing water extract followed by an ethanol + water extract, an ethanol extract followed by an ethanol + water extract, and an existing ethanol + water extract followed by an ethanol extract, all resulted in a less effective collagen-promoting effect than when using an ethanol + water extract continuously.

[0125] Therefore, the applicant kept the amount of ethanol added constant at 100 parts, adjusted the amount of water added, and, referring to the ratios of experimental groups D-1 to D-18, obtained experimental groups F1 to F-18. Type I collagen content was then tested. The results showed that with a constant ethanol content of 100 parts and an added amount of deionized water of 200-220 parts, the product's effect was significant, with type I collagen levels ranging from 107.89 to 108.23 ng / mL. The best effect was achieved when the added amount of deionized water was 113 parts (53% by mass). Further addition of deionized water resulted in a significant decrease in product effectiveness, even to the point of almost losing its effectiveness.

[0126] Example 14 Based on the composition for promoting endogenous collagen production in Example 11, the pretreatment is as follows: dried saffron is placed in an acetic acid aqueous solution so that the dried saffron is completely submerged in the acetic acid aqueous solution, soaked for 25 hours, and then rinsed with clean water to complete the pretreatment; the acetic acid aqueous solution is an acetic acid aqueous solution with pH=4.5.

[0127] Example 15 Based on the composition for promoting endogenous collagen production in Example 11, the pretreatment is as follows: dried saffron is placed in an acetic acid aqueous solution so that the dried saffron is completely submerged in the acetic acid aqueous solution, soaked for 30 hours, and then rinsed with clean water to complete the pretreatment; the acetic acid aqueous solution is an acetic acid aqueous solution with pH=5.0.

[0128] Example 16 Based on the composition for promoting endogenous collagen production in Example 11, the pretreatment is as follows: dried saffron is placed in an acetic acid aqueous solution so that the dried saffron is completely submerged in the acetic acid aqueous solution, soaked for 20 hours, and then rinsed with clean water to complete the pretreatment; the acetic acid aqueous solution is an acetic acid aqueous solution with pH=4.0.

[0129] The applicant discovered through research that pretreatment of saffron with a certain concentration of acetic acid aqueous solution can further enhance the efficacy of saffron extract. Taking Example 14 of the present invention as an example, the resulting composition can further increase the detection amount of type I collagen from 108.23 ng / mL in Example 11 to 109.67 ng / mL.

[0130] Comparative Example 14 Based on the composition for promoting endogenous collagen production in Example 11, the pretreatment is as follows: dried saffron is placed in a nitric acid aqueous solution so that the dried saffron is completely submerged in the nitric acid aqueous solution, soaked for 25 hours, and then rinsed with clean water to complete the pretreatment; the nitric acid aqueous solution is a nitric acid aqueous solution with pH=4.5.

[0131] Comparative Example 15 Based on the composition for promoting endogenous collagen production in Example 11, the pretreatment is as follows: dried saffron is placed in a hydrochloric acid aqueous solution so that the dried saffron is completely submerged in the hydrochloric acid aqueous solution, soaked for 25 hours, and then rinsed with clean water to complete the pretreatment; the hydrochloric acid aqueous solution is a hydrochloric acid aqueous solution with pH=4.5.

[0132] Comparative Example 16 Based on the composition for promoting endogenous collagen production in Example 11, the pretreatment is as follows: dried saffron is placed in a sulfuric acid aqueous solution so that the dried saffron is completely submerged in the sulfuric acid aqueous solution, soaked for 25 hours, and then rinsed with clean water to complete the pretreatment; the hydrochloric acid aqueous solution is a sulfuric acid aqueous solution with pH=4.5.

[0133] The applicant replaced the acetic acid aqueous solution with hydrochloric acid, nitric acid and sulfuric acid of the same pH, and found that the soaking effect was not good. The resulting composition did not increase the detection amount of type I collagen by less than 0.5 ng / mL compared with Example 1, and there was no significant increase.

[0134] Example 17 Based on the composition for promoting endogenous collagen production from Example 11, a microwave field of 575-615 MHz was continuously applied during step (II), reducing the extraction time to 30-40 min, preferably 35 min. For saffron extract, the application of a microwave field significantly improved extraction efficiency; however, no improvement in the efficacy of the composition was found by changing the microwave field.

[0135] Based on the above-described preferred embodiments of the present invention, and through the foregoing description, those skilled in the art can make various changes and modifications without departing from the inventive concept. The technical scope of this invention is not limited to the contents of the specification, but must be determined according to the scope of the claims.

Claims

1. A composition for promoting the production of endogenous collagen, characterized in that, include: The main components and auxiliary components, wherein the amount of auxiliary components added is 5-7% of the mass of the main components, wherein: the main components include: 100 parts silk fibroin and 10-50 parts peptides; the auxiliary components are: Panax notoginseng extract, or, by mass percentage, 63-67% Panax notoginseng extract and the balance being saffron extract. The peptides include: 15-20 parts carnosine, 8-12 parts tetrapeptide-3, 10-15 parts acetyl hexapeptide-8 and 10-20 parts arginine / lysine polypeptide; The preparation method of the Panax notoginseng extract includes: (1) Place the crushed Panax notoginseng into an extraction tank and add 7-10 times the weight of the first extract of Panax notoginseng; (2) In an environment of 30-40℃, the liquid phase components were circulated for 5-7 hours to obtain the first extraction product liquid; (3) Add 10-12 times the weight of the second extract of Panax notoginseng and circulate the extract at 95-100℃ for 2-4 hours; drain the liquid phase components and cool to obtain the second extract product liquid; (4) The first extraction product liquid and the second extraction product liquid are mixed to obtain an extraction mixture; (5) The extract mixture is freeze-dried to obtain the Panax notoginseng extract; By mass fractions, the first extract comprises: 30-35 parts deionized water and 100 parts ethanol; the second extract is deionized water.

2. The composition for promoting endogenous collagen production according to claim 1, characterized in that, The main components, by mass, include 100 parts silk fibroin and 18.75-42.85 parts peptides.

3. The composition for promoting endogenous collagen production according to claim 1, characterized in that, The saffron extract is obtained by ethanol extraction of dried saffron.

4. A mixture that promotes the production of endogenous collagen, characterized in that, The composition is prepared by mixing 0.01-2‰ of the composition with the balance of deionized water in a mass fraction of 1 / 3 to obtain the mixture that promotes the production of endogenous collagen; the composition is the composition that promotes the production of endogenous collagen according to any one of claims 1-3.

5. The application of a composition that promotes the production of endogenous collagen, characterized in that, The composition for promoting endogenous collagen production as described in any one of claims 1-3 may be used in the preparation of skin care products.

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