A cell active peptide composition with firming and anti-aging effects and its preparation method and application

Abstract

The present invention relates to the field of medicine and cosmetology technology, and in particular to a cell active peptide composition with firming and anti-aging effects, and a preparation method and application thereof. The composition includes acetyl tetrapeptide-9, acetyl tetrapeptide-3, palmitoyl tetrapeptide-7, dipeptide-4, tripeptide-1, carnosine, snake venom peptide, acetyl hexapeptide-8, NAD+, ceramide, arnica flower extract, resveratrol, glutathione, dipeptide-1, panthenol, glabridin, and decarboxylated carnosine hydrochloride. The composition can activate cell energy; combat skin aging factors such as photoaging, reactive oxygen, inflammation, and saccharification; enhance skin barrier; promote extracellular matrix synthesis, strengthen basement membrane, and increase dermal density; strengthen subcutaneous adipose tissue; relax neuromuscular junctions, and smooth wrinkles. The composition is applied to skin medicine and cosmetology, reduces various wrinkles such as static lines and dynamic lines, improves the firmness and elasticity of the skin, and has multi-link firming and anti-aging effects such as wrinkle removal, lifting, and improvement of sagging.

Description

Technical Field

[0001] The present invention relates to the field of medicine and cosmetic technology, and in particular to a cell active peptide composition with firming and anti-aging effects, and a preparation method and application thereof. Background Art

[0002] Skin aging is a complex and multifactorial process, including decreased epidermal hydration capacity, photoaging, environmental pollution, bad living habits, and psychological stress such as negative emotions and stress. One of the most significant signs is the increase in facial wrinkles. This phenomenon is a natural manifestation of the passage of time. Our facial expressions and repetitive muscle movements, especially excessive muscle stretching, are one of the factors that cause wrinkles. By adjusting the contraction of facial muscles, we can effectively reduce the formation of wrinkles.

[0003] There is another mechanism for the formation of skin wrinkles: with age, the extracellular matrix in the skin, including collagen, elastin, fibronectin, hyaluronic acid, etc., will age and even degrade. The interaction between these molecules forms irregular bridges, which reduces the support of the skin structure, causing the skin to become loose and wrinkled.

[0004] The decline in skin metabolic capacity is another important cause of aging. The metabolic function of skin cells may be abnormal, such as mitochondrial dysfunction, increased levels of reactive oxygen species (ROS), etc. These metabolic abnormalities will accelerate the process of skin aging. Nicotinamide adenine dinucleotide (NAD+) is a key molecule in cellular energy metabolism. Its level decreases with age and is closely related to the aging process. NAD+ is also a cofactor or substrate for a variety of enzymes and plays an important role in regulating cellular processes and functions. Therefore, restoring NAD+ levels has become one of the effective strategies to combat cellular aging.

[0005] However, the existing technology has certain defects in combating skin aging, especially in terms of high-efficiency anti-aging and skin tightening effects. Summary of the invention

[0006] The present invention aims at the above technical problems and thus proposes the scheme of the present invention, which includes a cell active peptide composition with firming and anti-aging effects and a preparation method and application thereof.

[0007] The raw materials used in the preparation of the cell active peptide composition with firming and anti-aging effects of the present invention include: 0.0001-1 part of ceramide, 0.0001-0.5 part of glutathione, 0.0001-1 part of resveratrol, 0.0001-1 part of dipeptide-10.0001-1 part of panthenol 0.0001-1 part of tripeptide-10.0001-1 part of glabridin 0.0001-1 part of dipeptide-40.0001-1 part of carnosine 0. 0.0001-1 part, decarboxylated carnosine hydrochloride 0.0001-1 part, palmitoyl tetrapeptide-7 0.0001-0.1 part, NAD+ 0.0001-1 part, acetyl tetrapeptide-9 0.0001-1 part, acetyl tetrapeptide-3 0.0001-1 part, acetyl hexapeptide-8 0.0001-0.5 part, snake venom peptide 0.0001-0.5 part, arnica flower extract 0.001-2 parts; measured by mass.

[0008] Among them, ceramide: forms a network structure, associates water molecules, enhances barrier function, improves skin dryness, regulates water and oil balance, and promotes skin repair.

[0009] Glutathione: It can resist the damage of ultraviolet rays to the human body, maintain the elasticity of the skin and prevent photoaging.

[0010] Dipeptide-4: can increase the skin's antioxidant defense ability and reduce the oxidation of skin lipids.

[0011] Palmitoyl Tetrapeptide-7: Reduces IL-6 secretion and reduces inflammation after UVB exposure.

[0012] NAD+: Also known as nicotinamide adenine dinucleotide, coenzyme I or nicotinamide dipurine nucleotide, it promotes cellular energy metabolism, activates the protein activity of the longevity protein Stir, promotes the TCA cycle and ATP production, protects mitochondria, repairs ultraviolet damage, and inhibits oxidative stress TNF-α inflammatory factors.

[0013] Resveratrol: It is a compound with powerful antioxidant effects. It can remove free radicals and reduce the damage of oxidative stress to the body. Resveratrol can also inhibit lipid peroxidation and regulate the activity of antioxidant-related enzymes, further enhancing its antioxidant and anti-free radical effects.

[0014] Acetyl tetrapeptide-9: stimulates the synthesis of perlecan.

[0015] Acetyl Tetrapeptide-3: Acts on fibroblasts, accelerates the synthesis of laminin, collagen III and VII, and repairs and strengthens the epidermis-dermis junction (DEJ).

[0016] Tripeptide-1: Stimulates collagen production, has wrinkle-reducing and anti-aging effects.

[0017] Acetyl Hexapeptide-8: It competes with SNAP-25 for its position in the SNARE complex, preventing the vesicles from effectively binding to the membrane to release acetylcholine, blocking muscle electrical impulse signals, inhibiting the release of neurotransmitters, and relaxing facial muscles, achieving the effect of smoothing dynamic lines, static lines and fine lines.

[0018] Snake venom peptide: dipeptide diaminobutyryl benzylamide diacetate, can act on the postsynaptic membrane, reversibly bind to nicotine acetylcholine receptors (mnAchR), competitively inhibit the absorption of acetylcholine, and block the nerve signal transmission on the acetylcholine receptors. Because the nicotine acetylcholine receptors in the muscles are blocked, the ion channels remain closed, no sodium ions are absorbed, and the muscle cells remain relaxed, thereby improving pseudo-wrinkles.

[0019] Arnica flower extract: Arnica flower extract is rich in flavonoids, anthocyanins and other compounds, and has excellent free radical scavenging and antioxidant effects, protecting the skin from free radical damage such as ultraviolet radiation and pollutants. It can also accelerate the growth and differentiation of human subcutaneous pre-adipocytes and make the skin plump. It promotes the proliferation of pre-adipocytes, promotes the storage of fat droplets by adipocytes, and increases the volume of adipocytes.

[0020] Furthermore, the ceramide may be present in any amount of 0.0005, 0.001, 0.002, 0.005, 0.01, 0.05, 0.1, 0.2, 0.5, 0.8 or 1 part by weight.

[0021] Furthermore, the mass fraction of glutathione can also be any numerical amount among 0.0005 parts, 0.001 parts, 0.002 parts, 0.005 parts, 0.01 parts, 0.05 parts, 0.1 parts, 0.2 parts, and 0.5 parts.

[0022] Furthermore, the mass fraction of the resveratrol can be any numerical amount among 0.0005, 0.001, 0.002, 0.005, 0.01, 0.05, 0.1, 0.2, 0.5, 0.8, and 1.

[0023] Furthermore, the dipeptide-1 part by mass can be any numerical amount among 0.0005 part, 0.001 part, 0.002 part, 0.005 part, 0.01 part, 0.05 part, 0.1 part, 0.2 part, 0.5 part, 0.08 part, and 1 part.

[0024] Furthermore, the mass fraction of panthenol can be any numerical amount among 0.0005 part, 0.001 part, 0.002 part, 0.005 part, 0.01 part, 0.05 part, 0.1 part, 0.2 part, 0.5 part, 0.08 part, and 1 part.

[0025] Furthermore, the tripeptide-1 part by mass can also be any numerical amount among 0.0005 part, 0.001 part, 0.002 part, 0.005 part, 0.01 part, 0.05 part, 0.1 part, 0.2 part, 0.5 part, 0.08 part, and 1 part.

[0026] Furthermore, the mass portion of the glabridin may be any numerical amount of 0.0005, 0.001, 0.002, 0.005, 0.01, 0.05, 0.1, 0.2, 0.5, 0.08, or 1.

[0027] Furthermore, the mass fraction of the dipeptide-4 may be any numerical amount among 0.0005, 0.001, 0.002, 0.005, 0.01, 0.05, 0.1, 0.2, 0.5, 0.08, and 1.

[0028] Furthermore, the carnosine may be in any amount of 0.0005, 0.001, 0.002, 0.005, 0.01, 0.05, 0.1, 0.2, 0.5, 0.08 or 1 part by weight.

[0029] Furthermore, the mass fraction of the decarboxylated carnosine hydrochloride can also be any numerical amount among 0.0005 part, 0.001 part, 0.002 part, 0.005 part, 0.01 part, 0.05 part, 0.1 part, 0.2 part, 0.5 part, 0.08 part and 1 part.

[0030] Furthermore, the mass fraction of the palmitoyl tetrapeptide-7 can be any numerical amount among 0.0005 parts, 0.001 parts, 0.002 parts, 0.005 parts, 0.01 parts, 0.05 parts, and 0.1 parts.

[0031] Furthermore, the mass fraction of NAD+ can also be any numerical amount among 0.0005 parts, 0.001 parts, 0.002 parts, 0.005 parts, 0.01 parts, 0.05 parts, and 0.1 parts.

[0032] Furthermore, the mass fraction of the acetyl tetrapeptide-9 can be any numerical amount among 0.0005, 0.001, 0.002, 0.005, 0.01, 0.05, 0.1, 0.2, 0.5, 0.08, and 1.

[0033] Furthermore, the mass fraction of the acetyl tetrapeptide-3 can also be any numerical amount among 0.0005 parts, 0.001 parts, 0.002 parts, 0.005 parts, 0.01 parts, 0.05 parts, 0.1 parts, 0.2 parts, 0.5 parts, 0.08 parts, and 1 part.

[0034] Furthermore, the mass fraction of the acetyl hexapeptide-8 may be any numerical amount among 0.0005, 0.001, 0.002, 0.005, 0.01, 0.05, 0.1, 0.2, and 0.5.

[0035] Furthermore, the mass fraction of the snake venom-like peptide can be any numerical amount among 0.0005 parts, 0.001 parts, 0.002 parts, 0.005 parts, 0.01 parts, 0.05 parts, 0.1 parts, 0.2 parts, and 0.5 parts.

[0036] Furthermore, the mass fraction of the Arnica montana flower extract can be any numerical amount selected from 0.001, 0.002, 0.005, 0.01, 0.05, 0.1, 0.2, 0.5, 0.8, 1, 1.5, 1.8, and 2.

[0037] Furthermore, the raw materials of the cell active peptide composition with firming and anti-aging effects include, by mass, 0.01-0.5 parts of ceramide, 0.01-0.4 parts of glutathione, 0.08-0.9 parts of resveratrol, 0.01-0.3 parts of dipeptide-1, 0.02-0.2 parts of panthenol, 0.01-0.2 parts of tripeptide-1, 0.01-0.3 parts of glabridin, and 0.02-0.2 parts of dipeptide-4. , carnosine 0.01-0.2 parts, decarboxylated carnosine hydrochloride 0.01-0.1 parts, palmitoyl tetrapeptide-7 0.01-0.1 parts, NAD+ 0.005-0.1 parts, acetyl tetrapeptide-9 0.01-0.2 parts, acetyl tetrapeptide-3 0.002-0.4 parts, acetyl hexapeptide-8 0.01-0.4 parts, snake venom peptide 0.005-0.1 parts, and arnica flower extract 0.2-1.8 parts.

[0038] Furthermore, the mass ratio of resveratrol to arnica flower extract is 1:2-4, 1:2.3-4 or 1:2.5-3.2.

[0039] Furthermore, the preparation method of the arnica flower extract includes: crushing the dried arnica flower, adding water of 8-40 times the mass of the crushed material, heating to 50-70°C and stirring for 2-5 hours; sterilizing, then cooling to 10-30°C, adding sugar, and inoculating fermentation bacteria composed of Lactobacillus delbrueckii subspecies bulgaricus CICC 6099 and Bacillus subtilis CICC24713 in a ratio of 2-5:1 of live bacteria to obtain a material, then fermenting at 30-40°C for 40-70 hours, filtering and ultrafiltration, concentrating and drying the ultrafiltrate to obtain the arnica flower extract.

[0040] Furthermore, the sugar includes at least one of glucose, sucrose, brown sugar, white sugar and maltose.

[0041] Furthermore, the mass ratio of the sugar in the material is 2-6%.

[0042] Furthermore, in the material, the total amount of inoculated bacteria is 6.7×10 8 -2.5×10 9 cfu / mL.

[0043] The Lactobacillus delbrueckii subspecies bulgaricus CICC 6099: strain number: CICC 6099; purchased from China Industrial Microbiological Culture Collection Administration Center. The Bacillus subtilis CICC 24713: strain number: CICC 24713; purchased from China Industrial Microbiological Culture Collection Administration Center.

[0044] Furthermore, the ultrafiltration is performed using an ultrafiltration membrane with a molecular weight cutoff of 1500 Da.

[0045] Furthermore, the composition raw materials also include 20-50 parts by weight of water and 5-20 parts by weight of polyol.

[0046] Furthermore, the polyol includes at least one of ethylene glycol, propylene glycol, polyethylene glycol, and butylene glycol.

[0047] On the other hand, the present invention also provides a method for preparing the composition, comprising the step of mixing the above raw materials.

[0048] Furthermore, the preparation method also includes a sterilization step.

[0049] In another aspect, the present invention also provides use of the composition in medicine and beauty.

[0050] Furthermore, the present invention also provides use of the composition in preparing a skin external product.

[0051] In another aspect, the present invention also provides use of the composition in preparing cosmetics or medicines.

[0052] Furthermore, the cosmetics include at least one of lotion, cream, ointment, facial mask, foundation, essence and lotion.

[0053] Furthermore, the medicine includes a medicine for external use on the skin.

[0054] On the other hand, the present invention also provides use of the composition in preparing medical cosmetic products.

[0055] On the other hand, the present invention also provides use of the composition in medical cosmetology for non-therapeutic purposes.

[0056] Beneficial effects of the present invention:

[0057] The composition prepared by the present invention using acetyl tetrapeptide-9, acetyl tetrapeptide-3, palmitoyl tetrapeptide-7, dipeptide-4, tripeptide-1, carnosine, snake venom peptide, acetyl hexapeptide-8, NAD+, ceramide, arnica flower extract, resveratrol, glutathione, dipeptide-1, panthenol, glabridin, decarboxylated carnosine hydrochloride, etc. can significantly improve the firmness and elasticity of the skin and has efficient firming and anti-aging effects.

[0058] The cell active peptide composition of the present invention can increase the skin elasticity ΔR2 to 0.0571 after being used for 2 months. The cell active peptide composition of the present invention can effectively tighten the skin and give the skin high elasticity.

[0059] After dilution, the cell active peptide composition of the present invention can still effectively remove free radicals. The efficiency of the diluted composition in removing ABTS free radicals and hydroxyl free radicals is 92.786% and 90.157% respectively. The composition of the present invention has excellent anti-aging effects and gives the skin a healthy state.

[0060] The formula system of the present invention uses resveratrol and arnica flower extract, which can give the skin better firming and free radical scavenging effects.

[0061] The combination of resveratrol and arnica flower extract of the present invention has better free radical scavenging effects than any one of resveratrol and arnica flower extract, and the combination of the two has a mutually reinforcing compound relationship. When the ratio of the two is within the range of 1:2-4, the effect of the composition shows a better situation.

[0062] The fermentation of the specific bacteria of the present invention to prepare the arnica extract has a certain influence on the effect of the composition; the use of Lactobacillus delbrueckii subspecies bulgaricus CICC 6099 and Bacillus subtilis CICC 24713 in the fermentation process can synergistically promote the free radical scavenging effect of the extract, giving the composition a better free radical scavenging effect; and when the ratio of the two is within the range of 2-5:1, the fermentation effect is better, and the free radical scavenging active ingredient produced has better performance. The selection of the specific bacteria of the present invention is better than the fermentation of other bacteria to prepare the arnica extract, and the effect produced is better.

[0063] The composition of the invention has excellent effect, is easy to prepare and has good application prospect. DETAILED DESCRIPTION

[0064] The present invention will be described below in conjunction with specific embodiments, and various effects of the present invention will be more clearly presented. It should be understood by those skilled in the art that these specific embodiments are used to illustrate the present invention, rather than to limit the present invention.

[0065] Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or prepared by existing methods.

[0066] 1. Preparation of Arnica Montana Flower Extract

[0067] 1. Arnica Flower Extract A

[0068] The preparation method is as follows:

[0069] The dried arnica flower is crushed, 25 times the mass of water of the crushed material is added, and the mixture is heated to 65° C. and stirred for 4 hours; sterilized, then cooled to 25° C., glucose is added, and fermentation bacteria consisting of Lactobacillus delbrueckii subspecies bulgaricus CICC6099 and Bacillus subtilis CICC 24713 are inoculated in a ratio of 4:1 of live bacteria to obtain a material, which is then anaerobically fermented at 37° C. for 50 hours, filtered using a filter membrane with a pore size of 1 micron, and the filtrate is ultrafiltered using an ultrafiltration membrane with a molecular weight cutoff of 1500 Da, and the ultrafiltrate is concentrated under reduced pressure at 60° C. to 13.7% of the mass of the ultrafiltrate, and freeze-dried at -40° C. to a water content of 3.26wt%, to obtain the arnica flower extract.

[0070] Among them, the mass concentration of glucose in the material is 4.27%, and the total amount of inoculated bacteria is 1.8×10 9 cfu / mL.

[0071] 2. Arnica Flower Extract B

[0072] The preparation method is as follows:

[0073] The dried arnica flower is crushed, 30 times the mass of water of the crushed material is added, and the mixture is heated to 62° C. and stirred for 3.8 hours; sterilized, then cooled to 25° C., glucose is added, and fermentation bacteria consisting of Lactobacillus delbrueckii subspecies bulgaricus CICC 6099 and Bacillus subtilis CICC 24713 are inoculated in a ratio of 2:1 of live bacteria to obtain a material, which is then anaerobically fermented at 38° C. for 40 hours, filtered using a filter membrane with a pore size of 1 micron, and the filtrate is ultrafiltered using an ultrafiltration membrane with a molecular weight cutoff of 1500 Da, and the ultrafiltrate is concentrated under reduced pressure at 60° C. to 12.4% of the mass of the ultrafiltrate, and freeze-dried at -40° C. to a water content of 3.17wt%, to obtain the arnica flower extract.

[0074] The mass concentration of glucose in the material is 5%, and the total amount of inoculated bacteria is 2×10 9 cfu / mL.

[0075] 3. Arnica montana flower extract C

[0076] The preparation method is as follows:

[0077] The dried arnica flower is crushed, 38 times the mass of water of the crushed material is added, and the mixture is heated to 65° C. and stirred for 5 hours; sterilized, then cooled to 25° C., glucose is added, and fermentation bacteria consisting of Lactobacillus delbrueckii subspecies bulgaricus CICC6099 and Bacillus subtilis CICC 24713 are inoculated in a ratio of 4:1 of live bacteria to obtain a material, which is then anaerobically fermented at 38° C. for 62 hours, filtered using a filter membrane with a pore size of 1 micron, and the filtrate is ultrafiltered using an ultrafiltration membrane with a molecular weight cutoff of 1500 Da, and the ultrafiltrate is concentrated under reduced pressure at 60° C. to 17.3% of the mass of the ultrafiltrate, and freeze-dried at -40° C. to a water content of 3.09wt%, to obtain the arnica flower extract.

[0078] Among them, the mass concentration of glucose in the material is 5.8%, and the total amount of inoculated bacteria is 1.5×10 9 cfu / mL.

[0079] 4. Arnica montana flower extract D

[0080] The preparation method is as follows:

[0081] The dried arnica flower is crushed, 25 times the mass of water of the crushed material is added, and the mixture is heated to 65° C. and stirred for 4 hours; sterilized, then cooled to 25° C., glucose is added to obtain a material, and then allowed to stand at 37° C. for 50 hours, filtered using a filter membrane with a pore size of 1 micron, and the filtrate is ultrafiltered using an ultrafiltration membrane with a molecular weight cutoff of 1500 Da, the ultrafiltrate is concentrated under reduced pressure at 60° C. to 13.7% of the mass of the ultrafiltrate, and freeze-dried at -40° C. to a water content of 3.26wt%, to obtain the arnica flower extract.

[0082] Wherein, the mass concentration of glucose in the material is 4.27%.

[0083] 5. Arnica montana flower extract E

[0084] The preparation method is as follows:

[0085] The dried arnica flower is crushed, 25 times the mass of water of the crushed material is added, and the mixture is heated to 65° C. and stirred for 4 hours; sterilized, then cooled to 25° C., glucose is added, and fermentation bacteria Lactobacillus delbrueckii subspecies bulgaricus CICC 6099 is inoculated to obtain a material, which is then anaerobically fermented at 37° C. for 50 hours, filtered using a filter membrane with a pore size of 1 micron, and the filtrate is ultrafiltered using an ultrafiltration membrane with a molecular weight cutoff of 1500 Da, and the ultrafiltrate is concentrated under reduced pressure at 60° C. to 13.7% of the mass of the ultrafiltrate, and freeze-dried at -40° C. to a water content of 3.26wt%, to obtain the arnica flower extract.

[0086] Among them, the mass concentration of glucose in the material is 4.27%, and the total amount of inoculated bacteria is 1.8×10 9 cfu / mL.

[0087] 6. Arnica montana flower extract F

[0088] The preparation method is as follows:

[0089] The dried arnica flower is crushed, 25 times the mass of water of the crushed material is added, and the mixture is heated to 65° C. and stirred for 4 hours; sterilized, then cooled to 25° C., glucose is added, and fermentation bacteria Bacillus subtilis CICC 24713 is inoculated to obtain a material, which is then anaerobically fermented at 37° C. for 50 hours, filtered using a filter membrane with a pore size of 1 micron, and the filtrate is ultrafiltered using an ultrafiltration membrane with a molecular weight cutoff of 1500 Da, and the ultrafiltrate is concentrated under reduced pressure at 60° C. to 13.7% of the mass of the ultrafiltrate, and freeze-dried at -40° C. to a water content of 3.26wt%, to obtain the arnica flower extract.

[0090] Among them, the mass concentration of glucose in the material is 4.27%, and the total amount of inoculated bacteria is 1.8×10 9 cfu / mL.

[0091] 7. Arnica montana flower extract G

[0092] The preparation method is as follows:

[0093] The dried arnica flower is crushed, 25 times the mass of water of the crushed material is added, and the mixture is heated to 65° C. and stirred for 4 hours; sterilized, then cooled to 25° C., glucose is added, and fermentation bacteria consisting of Lactobacillus delbrueckii subspecies bulgaricus CICC 6099 and Bacillus subtilis CICC 24713 are inoculated at a ratio of 0.5:1 of live bacteria to obtain a material, which is then anaerobically fermented at 37° C. for 50 hours, filtered using a filter membrane with a pore size of 1 micron, and the filtrate is ultrafiltered using an ultrafiltration membrane with a molecular weight cutoff of 1500 Da, and the ultrafiltrate is concentrated under reduced pressure at 60° C. to 13.7% of the mass of the ultrafiltrate, and freeze-dried at -40° C. to a water content of 3.26wt%, to obtain the arnica flower extract.

[0094] Among them, the mass concentration of glucose in the material is 4.27%, and the total amount of inoculated bacteria is 1.8×10 9 cfu / mL.

[0095] 8. Arnica montana flower extract H

[0096] The preparation method is as follows:

[0097] The dried arnica flower is crushed, 25 times the mass of water of the crushed material is added, and the mixture is heated to 65° C. and stirred for 4 hours; sterilized, then cooled to 25° C., glucose is added, and fermentation bacteria consisting of Lactobacillus delbrueckii subspecies bulgaricus CICC6099 and Bacillus subtilis CICC 24713 are inoculated in a ratio of 8:1 of live bacteria to obtain a material, which is then anaerobically fermented at 37° C. for 50 hours, filtered using a filter membrane with a pore size of 1 micron, and the filtrate is ultrafiltered using an ultrafiltration membrane with a molecular weight cutoff of 1500 Da, and the ultrafiltrate is concentrated under reduced pressure at 60° C. to 13.7% of the mass of the ultrafiltrate, and freeze-dried at -40° C. to a water content of 3.26wt%, to obtain the arnica flower extract.

[0098] Among them, the mass concentration of glucose in the material is 4.27%, and the total amount of inoculated bacteria is 1.8×10 9 cfu / mL.

[0099] 9. Arnica montana flower extract I

[0100] The preparation method is as follows:

[0101] The dried arnica flower is crushed, 25 times the mass of water of the crushed material is added, and the mixture is heated to 65° C. and stirred for 4 hours; sterilized, then cooled to 25° C., glucose is added, and fermentation bacteria Lactobacillus reuteri CICC 6132 (strain number: CICC6132, purchased from China Industrial Microbiological Culture Collection Center) are inoculated to obtain a material, which is then anaerobically fermented at 37° C. for 50 hours, filtered using a filter membrane with a pore size of 1 micron, and the filtrate is ultrafiltered using an ultrafiltration membrane with a molecular weight cutoff of 1500 Da, and the ultrafiltrate is concentrated under reduced pressure at 60° C. to 13.7% of the mass of the ultrafiltrate, and freeze-dried at -40° C. to a water content of 3.26wt%, to obtain the arnica flower extract.

[0102] Among them, the mass concentration of glucose in the material is 4.27%, and the total amount of inoculated bacteria is 1.8×10 9 cfu / mL.

[0103] 2. Preparation of the composition

[0104] The raw materials and weight parts of the composition are shown in Table 1.

[0105] Table 1: Raw material composition of the composition

[0106]

[0107] Compositions 1-B to 1-I are compositions obtained by replacing the Arnica montana flower extract A in Composition 1 with the Arnica montana flower extract BI prepared above.

[0108] The preparation method of all the above compositions comprises: placing various raw materials in a mixing container, stirring at 60 rpm for 0.5 hour, sterilizing, and obtaining.

[0109] 3. Effect Test

[0110] 1. Skin firming effect test

[0111] 64 female volunteers aged 30-45 with healthy skin were recruited and randomly divided into groups according to age, with 4 people in each group; each group of volunteers applied the above composition to their faces after washing their faces every night, with a dosage of 0.5g each time, and continued to use it for 2 months. Before use and 2 months after use, the skin elasticity of the volunteers' cheeks was tested. The specific test was to use the skin elasticity tester Cutometer, MPA580 to test the parameter R2 (the ratio of the rebound amount without negative pressure to the maximum pull-up amount with negative pressure) at 5 random locations on the cheeks as the basis for judging skin elasticity; among them, the closer the value of parameter R2 is to 1, the better the skin elasticity. The specific test results are shown in Table 2.

[0112] Table 2 Skin tightening effect test

[0113]

[0114]

[0115] According to the contents of Table 2, the change in skin elasticity △R2 of each group of volunteers after 2 months of use compared with before use was calculated, and the results are shown in Table 3.

[0116] Table 3: Changes in skin elasticity △R2

[0117]

[0118]

[0119] 2. Anti-aging effect test

[0120] The free radical scavenging effect of the above composition was tested to verify the anti-aging performance. The specific tests are as follows:

[0121] ABTS free radical scavenging ability

[0122] ABTS solution: Prepare ABTS into a 2 mmol / L solution with distilled water, then take 50 mL and mix it with 200 mL of 70 mmol / L K2S2O8 aqueous solution, and place it in the dark for 10 hours to obtain ABTS solution.

[0123] A 0.2 mol / L NaH2PO4 aqueous solution and a 0.2 mol / L Na2HPO4 aqueous solution were mixed in a volume ratio of 31:9 to obtain a buffer solution, and then the ABTS solution was diluted to an absorbance of 1.0 using the buffer solution to obtain a diluted ABTS solution. 1.2 mL of the dilution of the above composition (diluted to 20 wt% with deionized water) was taken and added to 1.8 mL of the diluted ABTS solution, mixed, allowed to stand for 15 minutes, and the absorbance at 734 nm was measured using a spectrophotometer. The scavenging rate γ of the ABTS free radical was calculated, where γ = 100% × (A0-A) / A0, A0 is the absorbance of the diluted ABTS solution at 734 nm, and A is the absorbance of the solution obtained by mixing the diluted ABTS solution and the composition at 734 nm.

[0124] Hydroxyl free radical scavenging ability:

[0125] Specific determination process: prepare 8.8mmol / L H2O2 solution, 9.0mmol / L ferrous sulfate solution and 9.0mmol / L salicylic acid solution with deionized water. Add 1mL 8.8mmol / L H2O2 solution, 1mL 9.0mmol / L ferrous sulfate solution and 1mL 9.0mmol / L salicylic acid solution to the container to obtain a control test solution for standby use. Mix 1mL 8.8mmol / L H2O2 solution, 1mL 9.0mmol / L ferrous sulfate solution and 1mL 9.0mmol / L salicylic acid solution with 2mL of the dilution (diluted to 20wt% with deionized water) of the above composition to obtain a test sample solution, and leave it for half an hour. Measure the absorbance of the control test solution and the test sample with a spectrophotometer at a wavelength of 515nm. The hydroxyl radical scavenging rate β was calculated, where β = 100% × (A1-A2) / A1, A1 is the absorbance of the control test solution at 515 nm, and A2 is the absorbance of the test sample solution at 515 nm.

[0126] The specific test results are shown in Table 4.

[0127] Table 4: Anti-aging effect test

[0128]

[0129]

[0130] It can be seen from the test results in Tables 2-4 that the cell active peptide composition of the present invention has excellent skin tightening and anti-aging effects.

[0131] The cell active peptide composition of the present invention can increase the skin elasticity ΔR2 to 0.0571 after being used for 2 months. The cell active peptide composition of the present invention can effectively tighten the skin and give the skin high elasticity.

[0132] After dilution, the cell active peptide composition of the present invention can still effectively remove free radicals. The efficiency of the diluted composition in removing ABTS free radicals and hydroxyl free radicals is 92.786% and 90.157% respectively. The composition of the present invention has excellent anti-aging effects and gives the skin a healthy state.

[0133] It can be seen from the test results of compositions 1 and 4 in the table that the use of resveratrol and arnica flower extract in the formula system of the present invention can give the skin better firming and free radical scavenging effects.

[0134] The test results of compositions 1, 5-6 show that the combination of resveratrol and arnica extract has better free radical scavenging effects than any one of resveratrol and arnica extract, and the combination of the two has a mutually reinforcing relationship. When the ratio of the two is within the range of 1:2-4, the effect of the composition shows better results; the above content can be obtained by combining the analysis of compositions 1, 7-8.

[0135] It can be seen from compositions 1, 1-D to 1-H that the fermentation of the specific bacteria of the present invention to prepare the Arnica montana extract has a certain influence on the effect of the composition; the use of Lactobacillus delbrueckii subspecies bulgaricus CICC 6099 and Bacillus subtilis CICC 24713 in the fermentation process can synergistically promote the free radical scavenging effect of the extract, giving the composition a better free radical scavenging effect; and when the ratio of the two is within the range of 2-5:1, the fermentation effect is better, and the free radical scavenging active ingredient produced has better performance. And from the test results of composition 1-I, it can be seen that the selection of the specific bacteria of the present invention produces a better effect than fermentation of other bacteria to prepare Arnica montana extract.

[0136] The raw materials used in the present invention, such as acetyl tetrapeptide-9, acetyl tetrapeptide-3, palmitoyl tetrapeptide-7, dipeptide-4, tripeptide-1, carnosine, snake venom peptide, acetyl hexapeptide-8, NAD+, ceramide, arnica extract, resveratrol, glutathione, dipeptide-1, panthenol, glabridin, decarboxylated carnosine hydrochloride, etc., can activate cell energy, fight against skin aging factors such as photoaging, active oxygen, inflammation, and glycation, strengthen the skin barrier, promote the synthesis of extracellular matrix, strengthen the basement membrane, and increase the density of the dermis; they can also strengthen subcutaneous fat tissue, relax neuromuscular junctions, and smooth wrinkles. The composition is applied to skin medicine and cosmetology, reduces various wrinkles such as static lines and dynamic lines, improves the firmness and elasticity of the skin, and has multi-link firming and anti-aging effects such as wrinkle removal, lifting, and improvement of sagging.

[0137] The composition provided by the present invention belongs to a cell active peptide composition with multi-link firming and anti-aging effects. By compounding a specific ratio of cell active peptides and synergists, the problems of skin aging and wrinkles in the epidermis, basement membrane layer, dermis, subcutaneous muscle and fat are solved, the skin energy and firmness are comprehensively improved, and skin wrinkles are reduced; thus achieving a multi-link firming and anti-aging effect.

[0138] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A firming and anti-aging cell active peptide composition, characterized in that: The invention comprises the following raw materials in parts by mass: 0.0001-1 part of ceramide, 0.0001-0.5 part of glutathione, 0.0001-1 part of resveratrol, 0.0001-1 part of dipeptide-1, 0.0001-1 part of panthenol, 0.0001-1 part of tripeptide-1, 0.0001-1 part of glabridin, 0.0001-1 part of dipeptide-4, 0.0001-1 part of carnosine, 0.0001-1 part of decarboxylated carnosine hydrochloride, 0.0001-0.1 part of palmitoyl tetrapeptide-7, 0.0001-1 part of NAD+, 0.0001-1 part of acetyl tetrapeptide-9, 0.0001-1 part of acetyl tetrapeptide-3, 0.0001-1 part of acetyl hexapeptide-8 0.0001-0.5 parts, snake venom peptide 0.0001-0.5 parts, arnica flower extract 0.001-2 parts, water 20-50 parts and polyol 5-20 parts; The mass ratio of resveratrol to arnica extract is 1:2-4; The preparation method of the arnica flower extract comprises: crushing dried arnica flowers, adding water in an amount of 8-40 times the mass of the crushed material, heating to 50-70°C and stirring for 2-5 hours; sterilizing, then cooling to 10-30°C, adding sugar, and inoculating fermentation bacteria consisting of Lactobacillus delbrueckii subspecies bulgaricus CICC 6099 and Bacillus subtilis CICC 24713 in a ratio of 2-5:1 to obtain a material, then fermenting at 30-40°C for 40-70 hours, filtering and ultrafiltration, concentrating and drying the ultrafiltrate to obtain the arnica flower extract; the total amount of inoculated bacteria in the material is 6.7×10 8 -2.5×10 9 cfu / mL.

2. The cell active peptide composition according to claim 1, characterized in that: In parts by mass, it includes the following raw materials: Ceramide 0.01-0.5 parts, glutathione 0.01-0.4 parts, resveratrol 0.08-0.9 parts, dipeptide-1 0.01-0.3 parts, panthenol 0.02-0.2 parts, tripeptide-1 0.01-0.2 parts, glabridin 0.01-0.3 parts, dipeptide-4 0.02-0.2 parts, carnosine 0.01-0.2 parts, decarboxylated carnosine hydrochloride 0.01-0.1 parts, palmitoyl tetrapeptide-7 0.01-0.1 parts, NAD+ 0.005-0.1 parts, acetyl tetrapeptide-9 0.01-0.2 parts, acetyl tetrapeptide-3 0.002-0.4 parts, acetyl hexapeptide-8 0.01-0.4 parts, 0.005-0.1 parts of snake venom peptide, 0.2-1.8 parts of arnica flower extract, 20-50 parts of water and 5-20 parts of polyol.

3. The cell active peptide composition according to claim 1 or 2, characterized in that: The polyol includes at least one of ethylene glycol, propylene glycol, polyethylene glycol, and butylene glycol.

4. A method for preparing the cell active peptide composition according to any one of claims 1 to 3, characterized in that: The process comprises the steps of mixing the raw materials.

5. Use of the cell active peptide composition according to any one of claims 1 to 3 in the preparation of external skin products.

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