A composition with nerve-soothing effect and its application

Abstract

This invention discloses a composition with a neuro-soothing effect and its application, wherein the composition comprises acetyl hexapeptide-1, and further comprises capsaicin or calvanolactone. This invention also discloses the application of the composition with a neuro-soothing effect in the preparation of cosmetics or skin care products. This invention features enhanced neuro-soothing effects and reduced material costs.

Description

Technical Field

[0001] This invention relates to a composition containing acetyl hexapeptide-1, and more particularly to a composition having a neuroleptic effect and its application. Background Technology

[0002] With the continuous improvement of living standards, how to alleviate wrinkles has always been a key research topic in the cosmetics industry. Skin is affected by changes in the external environment and internal factors, leading to aging and the appearance of wrinkles, sagging, and changes in skin tone, with wrinkles being one of the most obvious signs. The prolonged and frequent contraction of facial muscles gradually forms skin lines, eventually resulting in dynamic wrinkles. The contraction of facial muscles mainly relies on the transmission of nerve signals, a complex electrochemical reaction process.

[0003] Based on current research into the mechanisms of dynamic wrinkles, existing technologies primarily work by blocking neurotransmitter transmission, thereby relaxing muscles and achieving a soothing and anti-wrinkle effect. Currently, commonly used anti-wrinkle substances on the market include neurotransmitter peptides, plant-based active ingredients, and enzymes. However, peptide raw materials suffer from high production costs and difficulty in mass production. Furthermore, high concentrations make them difficult to combine with other ingredients, increasing R&D costs for companies. In addition, synaptic signal transmission is a complex dynamic process; a single ingredient has limited target sites and pathways. Using a small amount of a single ingredient yields poor results, while using a large amount increases costs. Summary of the Invention

[0004] The purpose of this invention is to provide a composition with a nerve-relieving effect and its application. This invention features enhanced nerve-relieving effect and reduced material costs.

[0005] The technical solution of the present invention is a composition with a nerve-soothing effect, comprising acetyl hexapeptide-1, and further comprising capsaicin or calvanolide.

[0006] In the aforementioned composition with a neuro-soothing effect, the composition comprises acetyl hexapeptide-1 and capsaicin, wherein the mass ratio of acetyl hexapeptide-1 to capsaicin is (1-4):(1.25-5).

[0007] In the aforementioned composition with a neuroleptic effect, the composition comprises acetyl hexapeptide-1 and capsaicin, wherein the concentration of acetyl hexapeptide-1 is 1 ppm to 4 ppm and the concentration of capsaicin is 1.25 ppm to 5 ppm.

[0008] In the aforementioned composition with a neuroleptic effect, the composition comprises acetyl hexapeptide-1 and capsaicin, wherein the concentration of acetyl hexapeptide-1 is 4 ppm and the concentration of capsaicin is 1.25 ppm to 5 ppm.

[0009] In the aforementioned composition with a neuroleptic effect, the composition comprises acetyl hexapeptide-1 and capsaicin, wherein the concentration of acetyl hexapeptide-1 is 1 ppm and the concentration of capsaicin is 1.25 ppm.

[0010] In the aforementioned composition with a neuroleptic effect, the composition comprises acetyl hexapeptide-1 and calvaline, wherein the mass ratio of acetyl hexapeptide-1 to calvaline is (0.5–10):(3.13–6.25).

[0011] In the aforementioned composition with a neuroleptic effect, the composition comprises acetyl hexapeptide-1 and calvanolide, wherein the concentration of acetyl hexapeptide-1 is 0.5 ppm to 10 ppm and the concentration of calvanolide is 3.13 ppm to 6.25 ppm.

[0012] In the aforementioned composition with a neuroleptic effect, the composition comprises acetyl hexapeptide-1 and calvanolide, wherein the concentration of acetyl hexapeptide-1 is 10 ppm and the concentration of calvanolide is 3.13 ppm to 6.25 ppm.

[0013] In the aforementioned composition with a neuroleptic effect, the composition comprises acetyl hexapeptide-1 and calvanolide, wherein the concentration of acetyl hexapeptide-1 is 0.5 ppm and the concentration of calvanolide is 3.13 ppm.

[0014] The present invention also discloses the application of the above-mentioned composition with nerve-soothing effect in the preparation of cosmetics or skin care products.

[0015] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0016] This invention combines acetyl hexapeptide-1, which is costly to produce and difficult to obtain, with calvanolide or capsaicin, an active monomer, which is cost-effective and easier to obtain. Acetyl hexapeptide-1 acts on the presynaptic membrane to soothe nerves by inhibiting the release of neurotransmitters, while calvanolide or capsaicin acts on the postsynaptic membrane to soothe nerves by blocking ion channels and inhibiting the action potentials generated by neurons.

[0017] The compositions of this invention exert a neuro-soothing effect through different mechanisms of action, which not only overcomes the deficiency of single-function bioactive substances, but also achieves multi-target action and synergistic effect. Moreover, the combined effect is far greater than the neuro-soothing effect of a single component at the same concentration, giving full play to the efficacy of peptides and improving the bioavailability of single components. It can reduce the amount of active ingredients while ensuring the same effect, which greatly reduces material costs. Detailed Implementation

[0018] A composition having a neuroleptic effect includes acetyl hexapeptide-1, and also includes capsaicin or calvanolide.

[0019] When the composition includes acetyl hexapeptide-1 and capsaicin, the mass ratio of acetyl hexapeptide-1 to capsaicin is (1-4):(1.25-5).

[0020] When the composition includes acetyl hexapeptide-1 and calvalactone, the mass ratio of acetyl hexapeptide-1 to calvalactone is (0.5–10):(3.13–6.25).

[0021] The present invention will be further described below with reference to embodiments, but these embodiments are not intended to limit the scope of the invention.

[0022] Example 1:

[0023] A composition having a neuroleptic effect comprises acetyl hexapeptide-1 at a concentration of 4 ppm and capsaicin at a concentration of 5 ppm.

[0024] Example 2:

[0025] A composition having a neuroleptic effect comprises acetyl hexapeptide-1 at a concentration of 4 ppm and capsaicin at a concentration of 2.5 ppm.

[0026] Example 3:

[0027] A composition having a neuroleptic effect comprises acetyl hexapeptide-1 at a concentration of 4 ppm and capsaicin at a concentration of 1.25 ppm.

[0028] Example 4:

[0029] A composition having a neuroleptic effect comprises acetyl hexapeptide-1 at a concentration of 1 ppm and capsaicin at a concentration of 1.25 ppm.

[0030] Example 5:

[0031] A composition having a neuroleptic effect comprises acetyl hexapeptide-1 at a concentration of 10 ppm and calvalactone at a concentration of 6.25 ppm.

[0032] Example 6:

[0033] A composition having a neuroleptic effect comprises acetyl hexapeptide-1 at a concentration of 0.5 ppm and calvalactone at a concentration of 3.13 ppm.

[0034] Comparative Example 1: The component with a neuroleptic effect is acetyl hexapeptide-1. In Comparative Examples 1-1 to 1-5, the concentrations of acetyl hexapeptide-1 were 10 ppm, 5 ppm, 2.5 ppm and 1.25 ppm, respectively.

[0035] Comparative Example 2: The component with a nerve-soothing effect is capsaicin. In Comparative Examples 2-1 to 2-3, the concentrations of capsaicin were 5 ppm, 2.5 ppm, and 1.25 ppm, respectively.

[0036] Comparative Example 3: The component with a neuroleptic effect is calvanolide. In Comparative Examples 3-1 and 3-2, the concentrations of calvanolide were 6.25 ppm and 3.13 ppm, respectively.

[0037] The specific components of each embodiment and comparative example are shown in Table 1.

[0038] Table 1. Specific ingredient list for each embodiment and comparative example

[0039]

[0040]

[0041] Zebrafish embryo seizure relief test:

[0042] 1.1 Experimental Principle:

[0043] Zebrafish possess neuroregulatory mechanisms similar to those in humans and have been widely used in the research and testing of neuroactive drugs. Through zebrafish embryonic behavior, instruments were used to record and software to measure the activity levels of zebrafish embryos. Changes in embryonic activity levels in the test drug-treated group and the control group were compared in a PTZ-induced seizure model, and the zebrafish neurorelaxation rate was calculated.

[0044] 1.2 Experimental Instruments and Equipment:

[0045] Zebrafish farming equipment, pH meter, dissolved oxygen meter, salinity meter, conductivity meter, analytical balance, microscope (with photographic system, stereo microscope with maximum magnification greater than 80x), 96-well plate, petri dishes, incubator, zebrafish motion recorder, brine shrimp incubator, pipettes, centrifuge tubes, beakers, volumetric flasks, vortex mixer, ultrasonic water bath, centrifuge, low temperature freezer, adjustable pipettes and pipette tips.

[0046] 1.3 Experimental Reagents:

[0047] Water (compliant with GB / T6682), methanol, dimethyl sulfoxide, ethanol and other cosolvents (analytical grade), water for broodstock aquaculture, Artemia salina eggs, fish embryo culture medium (prepared by dissolving 2940 mg anhydrous calcium chloride, 1233 mg magnesium sulfate heptahydrate, 630 mg sodium bicarbonate, and 55 mg potassium chloride in 10 L of water, pH 6.5–8.5, all chemicals are analytical grade), pentylenetetrazole (PTZ) stock solution (362 mM PTZ stock solution prepared from fish embryo culture medium and frozen).

[0048] 1.4 Experimental subjects:

[0049] Spawning tests were conducted using wild-type AB strain zebrafish (Daniorerio) from reliable sources (such as the China National Zebrafish Resource Center). The broodstock exhibited good reproductive capacity and were 6–12 months old. Lateral crosses were used to maintain genetic diversity, and purebred broodstock were replaced with a new batch after five generations. The broodstock showed no obvious signs of infection or disease and had not undergone drug treatment within the past two months. Prior to the spawning tests, the broodstock were acclimatized to the laboratory for at least 14 days after being introduced.

[0050] 1.5 Cultivation and treatment of experimental subjects:

[0051] The ideal water temperature for aquaculture is between 26℃ and 28.5℃, and the indoor temperature should be between 20℃ and 25℃. The stocking density should be controlled at 1-2 fish per liter of water. Provide a fixed 12-16 hours of light daily and maintain a good filtration system. Feed at least twice a day, including at least one feeding of Artemiasalina larvae, with feeding intervals of at least 3 hours to avoid overfeeding, which could negatively impact water quality and cleanliness.

[0052] Male and female zebrafish were placed in the spawning tank at a 2:1 ratio, 1-2 hours before the lights were turned off the day before spawning. Since zebrafish occasionally do not spawn, multiple spawning tanks were prepared as backups. To avoid genetic bias, eggs collected from at least three spawning tanks were mixed and then selected for later use. After the eggs were removed from the collection box, the embryos were cleaned with embryo culture medium. Healthy zebrafish embryos were selected and cultured at 28℃±1℃ at a density of no more than one embryo per 200μL of embryo culture medium until 5 days post-fertilization.

[0053] 1.6 Experimental Methods:

[0054] Experimental materials: Healthy zebrafish embryos, 5 days after fertilization, were selected.

[0055] Blank control group: 16 fish embryos were randomly selected and placed into a 30 mm culture dish, and 3.6 mL of fish embryo culture medium was added;

[0056] Model control group: 16 fish embryos were randomly selected and placed in a 30 mm culture dish, and 3.6 mL of fish embryo culture medium was added.

[0057] Test substance treatment group: 16 fish embryos were randomly selected and placed in a 30 mm culture dish, and 3.6 mL of test substance solution (i.e., the composition of the example or comparative example) was added.

[0058] The blank control group, model control group, and test substance treatment group were placed in a constant temperature incubator at 28℃±1℃ for 2 hours. Then, 76 μL of fish embryo culture medium was added to the blank control group, and 76 μL of LPTZ stock solution was added to the model control group and the test substance treatment group.

[0059] Embryo activity detection: Zebrafish embryos from the blank control group, model control group, and test substance treatment group were transferred into 96-well plates, with one embryo and 0.2 mL of the corresponding culture medium in each well. After sample addition, the 96 wells were transferred to a zebrafish motion recorder and allowed to acclimatize for 40 min. The activity of each zebrafish embryo was then recorded over 20 min using infrared tracking software.

[0060] Replace 180 μL of the solution in each well with fish embryo culture medium, and then place the wells in a constant temperature incubator at 28℃±1℃ for 24h±2h before counting the fish embryo survival rate.

[0061] 1.7 Data and Result Calculation:

[0062] Zebrafish neurorelaxation rate = zebrafish embryonic activity reduction rate (S / MB) * 100%; where S is the average embryonic activity in the test substance treatment group; B is the average embryonic activity in the blank control group; and M is the average embryonic activity in the model control group.

[0063] The nerve-soothing effect was evaluated by the reduction rate of swimming distance in zebrafish; the greater the reduction rate, the better the nerve-soothing effect.

[0064] If the swimming distance shortening rate in the calculation results is negative, it indicates that at the added concentration of the substance, there is no nerve-soothing effect, but rather a certain degree of nerve stimulation.

[0065] The neuro-soothing improvement rate of the composition relative to the single component is equal to Sab-Sa or Sab-Sb; where Sab is the neuro-soothing rate of the example; and Sa and Sb are the neuro-soothing rates of the single components of the same concentration in the comparative example.

[0066] In a PTZ-induced zebrafish embryonic seizure model, samples from various embodiments and comparative examples were added. The activity levels of the zebrafish embryos were recorded using detection instruments and measured using software. Changes in embryonic activity levels were compared between the model control group and the test substance treatment group. The activity-induced neuroleptic rate was calculated to evaluate the anticonvulsant and neuroleptic efficacy of the samples. The experimental results are shown in Table 2.

[0067] Table 2. Nerve Relaxation Rate of Various Embodiments and Comparative Examples

[0068]

[0069] As shown in Comparative Examples 1-1 to 1-5, in the PTZ-induced zebrafish embryonic seizure model, when the concentration of acetyl hexapeptide-1 was 10 ppm (Comparative Example 1-1), it reduced the activity of zebrafish embryos and had a good neurosoothing effect. When the concentration of acetyl hexapeptide-1 was 0.5–4 ppm (Comparative Examples 1-2–1-5), it had no neurosoothing effect on zebrafish and instead had a stimulating effect.

[0070] As shown in Comparative Examples 2-1 to 2-3, in the PTZ-induced zebrafish embryonic convulsion model, when the concentration of capsaicin was 1.25 ppm, 2.5 ppm, and 5 ppm, the reduction in zebrafish embryonic activity was not significant, and the effect on soothing the zebrafish nerves was poor.

[0071] As shown in Comparative Examples 3-1 to 3-2, in the PTZ-induced zebrafish embryonic convulsion model, the addition concentrations of calvaline at 6.25 ppm and 3.13 ppm did not significantly reduce the activity of zebrafish embryos and had a poor effect on soothing the nerves of zebrafish.

[0072] As shown in Examples 1-4, in the PTZ-induced zebrafish embryonic convulsion model, the addition of acetyl hexapeptide-1 at concentrations of 0.5 ppm, 1 ppm, 2 ppm, and 4 ppm, respectively, to capsaicin at concentrations of 1.25 ppm, 2.5 ppm, and 5 ppm, effectively reduced the activity of zebrafish embryos and significantly increased the neurosoothing rate. This was far superior to the neurosoothing effects of single components at corresponding concentrations and single components with similar total concentrations, indicating that acetyl hexapeptide-1 and capsaicin can have a synergistic effect in neurosoothing.

[0073] As shown in Examples 5 and 6, in the PTZ-induced zebrafish embryonic seizure model, the formulations of acetyl hexapeptide-1 at concentrations of 0.5 ppm and 10 ppm with calvanolide at concentrations of 3.13 ppm and 6.25 ppm, respectively, in Examples 5 and 6, effectively reduced zebrafish embryonic activity and increased the neurosoothing effect. These effects were significantly superior to those of single components at the corresponding concentrations and single components with similar total concentrations, indicating that acetyl hexapeptide-1 and calvanolide have a synergistic effect in neurosoothing.

[0074] In summary, the combination of acetyl hexapeptide-1, capsaicin, and calvanolactone at certain concentrations effectively reduces zebrafish embryonic activity and enhances the neuro-soothing effect in zebrafish, significantly outperforming the effects of the individual components and demonstrating a synergistic effect. Even when the total concentration of the individual components and the combined composition is approximately equal, it is difficult to achieve the same soothing effect as the combined composition. Therefore, the synergistic effect of the composition presented in this application is significant; furthermore, it saves materials and greatly reduces costs while achieving the same effect.

Claims

1. A composition with a nerve-soothing effect for use in the preparation of cosmetics, characterized in that: The composition includes acetyl hexapeptide-1, and also includes capsaicin or calvanolide; the composition includes acetyl hexapeptide-1 and capsaicin, with a mass ratio of acetyl hexapeptide-1 to capsaicin of (1~4):(1.25~5); the composition includes acetyl hexapeptide-1 and calvanolide, with a mass ratio of acetyl hexapeptide-1 to calvanolide of (0.5~10):(3.13~6.25).

2. The composition with a nerve-soothing effect for preparing cosmetics according to claim 1, characterized in that: The composition comprises acetyl hexapeptide-1 and capsaicin, wherein the concentration of acetyl hexapeptide-1 is 1 ppm to 4 ppm and the concentration of capsaicin is 1.25 ppm to 5 ppm.

3. A composition with a nerve-soothing effect for preparing cosmetics according to claim 2, characterized in that: The composition comprises acetyl hexapeptide-1 and capsaicin, wherein the concentration of acetyl hexapeptide-1 is 4 ppm and the concentration of capsaicin is 1.25 ppm to 5 ppm.

4. A composition with a nerve-soothing effect for preparing cosmetics according to claim 2, characterized in that: The composition comprises acetyl hexapeptide-1 and capsaicin, wherein the concentration of acetyl hexapeptide-1 is 1 ppm and the concentration of capsaicin is 1.25 ppm.

5. A composition with a nerve-soothing effect for preparing cosmetics according to claim 1, characterized in that: The composition comprises acetyl hexapeptide-1 and calvalactone, wherein the concentration of acetyl hexapeptide-1 is 0.5 ppm to 10 ppm and the concentration of calvalactone is 3.13 ppm to 6.25 ppm.

6. A composition with a nerve-soothing effect for preparing cosmetics according to claim 5, characterized in that: The composition comprises acetyl hexapeptide-1 and calvalactone, wherein the concentration of acetyl hexapeptide-1 is 10 ppm and the concentration of calvalactone is 3.13 ppm to 6.25 ppm.

7. A composition with a nerve-soothing effect for preparing cosmetics according to claim 5, characterized in that: The composition comprises acetyl hexapeptide-1 and calvalactone, wherein the concentration of acetyl hexapeptide-1 is 0.5 ppm and the concentration of calvalactone is 3.13 ppm.

8. The use of the composition with a neuro-soothing effect according to any one of claims 1-7 in the preparation of cosmetics.

9. The application according to claim 8, characterized in that: The cosmetics mentioned are skincare products.

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