Recombinant human elastin coffee peptide composition and application thereof

Abstract

The invention relates to the field of daily chemistry, is suitable for the field of scalp, hair and skin care, and provides a composition of recombinant human elastin peptide and Yunnan Arabic Coffee polypeptide and application of the composition. The innovative combination aims to radically relieve scalp discomfort and deeply relieve the problems of scalp redness, sensitivity and the like, meanwhile, disulfide bonds of hair are protected through a unique action mechanism, the condition of hair frizziness is remarkably relieved, the hair is recovered and kept in an ideal smooth state, and a brand new solution is brought to the field of scalp and hair care.

Description

Technical Field

[0001] The invention relates to the field of daily chemicals, is applicable to the field of scalp, hair and skin care, and particularly relates to a recombinant human elastin coffee peptide composition and application thereof. Background Art

[0002] With the continuous changes in people's lifestyles and the increasing complexity of environmental factors, scalp and hair care has become an important area of ​​concern. The significantly accelerated pace of life has put people under a long-term high-pressure state, which directly affects the body's normal metabolism and endocrine balance, which in turn is reflected in the health of the scalp and hair. The continuous increase in environmental pressure, such as air pollution and ultraviolet radiation, has caused direct damage to the scalp and hair. Frequent hair treatments, including perming, dyeing, straightening and other chemical operations, although satisfying people's pursuit of beauty, have inevitably caused damage to the structure of the hair and the ecological environment of the scalp.

[0003] In terms of the scalp, many people face a variety of problems due to the combined effects of the above factors. Scalp discomfort, such as itching, tightness, pain, inflammation, sensitive skin and other symptoms frequently occur, seriously affecting the quality of life. Redness and sensitivity are even more common. These are not only superficial symptoms, but may also be external manifestations of inflammation or damage to the deep tissues of the scalp. If not treated and cared for in time, it is likely to cause more serious scalp diseases, such as seborrheic dermatitis, scalp eczema, etc., which will affect the health of hair follicles and lead to serious consequences such as hair loss.

[0004] In terms of hair, the proteins that make up hair are mainly keratin and keratin-related proteins, which interact to form a complex network structure, giving hair good rigidity and toughness. Cysteine ​​accounts for 11-18% of keratin. The disulfide bonds formed between cysteine ​​molecules can enhance the strength of hair and give hair tough physical properties. Factors such as chemical treatment, heat damage, and mechanical friction during daily combing frequently damage the disulfide bonds in hair. Disulfide bonds play a vital role in maintaining the structure and morphology of hair. Damage to disulfide bonds can lead to structural instability of hair fibers, making hair frizzy, dry, and easy to break, losing its original smooth texture and luster. Hair care products can help consumers keep their hair healthy and clean and protect their hair from damage.

[0005] In terms of hair and scalp care, most current technologies can only solve some skin and scalp hair problems in a targeted manner. It is difficult to effectively and fundamentally reduce stubborn symptoms such as scalp redness and skin sensitivity, while at the same time protecting the hair's disulfide bonds, improving hair frizziness and enhancing its smoothness. It is difficult to achieve ideal care effects to meet consumer needs. Summary of the invention

[0006] The purpose of the present invention is to use the recombinant human elastin peptide and Yunnan small-grain coffee polypeptide composition in the field of scalp and hair care for the first time, and verify that the composition has anti-inflammatory biological activity by cell biological methods.

[0007] Another object of the present invention is to provide a Yunnan Arabica coffee polypeptide prepared by an enzymatic method.

[0008] Another object of the present invention is to provide a recombinant human elastin peptide and Yunnan Arabica coffee polypeptide composition having the effect of improving hair combing.

[0009] Another object of the present invention is to provide a recombinant human elastin peptide and Yunnan Arabica coffee polypeptide composition having anti-inflammatory effects.

[0010] Another object of the present invention is to predict the disulfide bond protection effect of the recombinant human elastin peptide and Yunnan Arabica coffee polypeptide composition by computer simulation.

[0011] In order to solve the above technical problems, the present invention adopts the following technical solutions: A composition comprises recombinant human elastin peptide and polypeptide derived from Arabica coffee (Coffea arabica L.) residues.

[0012] The recombinant human elastin and the polypeptide derived from Arabica coffee grounds are mixed to obtain a recombinant human elastin coffee peptide composition, wherein the elastin peptide and the polypeptide derived from Arabica coffee grounds are combined at a mass ratio of 1:100.

[0013] Preferably, the elastin peptide comprises a recombinant human elastin peptide; and the polypeptide derived from Arabica coffee grounds comprises a Yunnan Arabica coffee polypeptide.

[0014] Preferably, the recombinant human elastin peptide may be the recombinant human elastin peptide described in patent CN118290568A.

[0015] Preferably, the preparation of the Yunnan arabica coffee polypeptide comprises: using Yunnan arabica coffee grounds as raw material, reacting with protease, and obtaining the Yunnan arabica coffee polypeptide.

[0016] Preferably, the protease comprises ficin.

[0017] Preferably, the amino acid sequence of the Yunnan Arabica coffee polypeptide is shown in the sequence listing as SEQ ID NO: 1.

[0018] An application of the composition is used for one or more of hair care and scalp care.

[0019] Preferably, it is used to prepare a preparation for improving the combing performance of hair strands.

[0020] Preferably, the improving combing performance of the hair bundle includes one or more of improving the dry combing property of the hair bundle and improving the friction property of the hair bundle.

[0021] Preferably, it is used for preparing anti-inflammatory preparations.

[0022] Preferably, it is used for preparing a preparation for inhibiting the inflammatory factor IL-1α.

[0023] A preparation obtained by the above application.

[0024] Preferably, the preparation comprises one or more of shampoo and hair serum.

[0025] Preferably, in the preparation, the mass percentage of the composition includes 0.05% to 10%, preferably 0.05% to 2%.

[0026] The implementation of the present invention has the following beneficial effects: The present invention provides a composition of recombinant human elastin peptide and Yunnan small-grain coffee polypeptide and its application. This innovative combination fundamentally relieves scalp discomfort, deeply alleviates problems such as scalp redness and sensitivity, and at the same time protects the disulfide bonds of the hair through a unique mechanism of action, significantly reduces the frizziness of the hair, restores the hair to an ideal state of smoothness, and brings a new solution to the field of scalp and hair care. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 The results of the dry combing work difference test between the control group and the sample group are shown, where "**" indicates a significant difference, 0.001≤P<0.01.

[0028] Figure 2 The difference test results of friction work between the control group and the sample group are shown, where "*" indicates a significant difference, 0.01≤P<0.05.

[0029] Figure 3 The effects of different concentrations of LFT02 samples on the expression of inflammatory factor IL-1α induced by SLS are shown.

[0030] Figure 4 Schematic diagram of the covering effect of elastin fragments on keratin disulfide bond fragments.

[0031] Figure 5 Schematic diagram of the covering effect of KKAGNQGFEYVA in Arabica coffee peptide on the disulfide bond region in keratin. DETAILED DESCRIPTION

[0032] In order to make the objectives, technical solutions and advantages of the present invention more clear, the present invention will be further described in detail below with reference to the accompanying drawings.

[0033] Example 1 Enzymatic preparation of Yunnan Arabica coffee peptides The materials described in the experiment are shown in Table 1: Table 1

[0034] The crushed Yunnan Arabica coffee grounds (Latin name: Coffea Arabica) were selected as raw materials, and water was added to mix the ingredients so that the concentration reached 20% (weight ratio). The temperature was adjusted to 65°C and then 0.008% of ficin was added. The experimental reaction lasted for 2 hours, and low-speed stirring was maintained during the reaction. After the reaction, the mixture was boiled and kept warm for 10 minutes to inactivate the enzyme. After the inactivation of the enzyme, the mixture was centrifuged at 8000r / min for 10 minutes, and the supernatant was collected to obtain the Yunnan Arabica coffee polypeptide prepared by the enzymatic method.

[0035] Example 2 Yunnan Arabica coffee peptide improves hair combing effect Test materials and instruments: Slightly damaged cylindrical (Asian) hair tresses (3g, 17cmx3g, free 15cm).

[0036] 14% sodium lauryl sulfate (SLS) solution in water.

[0037] The formula of the control group is shown in Table 2, and the formula of the sample group is shown in Table 3.

[0038] Flat narrow-tooth comb: The tooth density is moderate (tooth spacing 0.9~1.1mm), the tooth length is 2.0~3.0cm, and the comb length is not less than 10cm.

[0039] Hair detangling device: fibra.one (Dia-Stron Limited).

[0040] Electronic balance: FA2004 (0.0001g).

[0041] Table 2 - Control Group Formulation

[0042] Table 3 - Sample set formulation

[0043] COFFEA ARABICA seed extract is a source of Coffea arabica L. grounds.

[0044] Hair strand pretreatment: Wet and rinse the hair with clean water, take 1mL of 14% sodium dodecyl sulfate (SLS) aqueous solution and evenly apply it on the surface of the hair, gently rub until rich foam lasts for 60s, rinse at a water temperature of 37.0±1.0℃ and a water flow rate of 4L / min, wash the hair twice with 14% SLS aqueous solution, comb the hair with a flat narrow-tooth comb for 10 times on each side after rinsing, and hang the hair in a constant temperature and humidity chamber at a temperature of 21±1℃ and a relative humidity of 50±10%RH for 16-24 hours.

[0045] Dry hair combability: 1. Select 10 hair bundles after pretreatment and randomly divide them into 2 groups, 5 bundles in the sample group and 5 bundles in the control group; 2. Initial value test: Use fibra.one to test the combability of hair bundles. Test each hair bundle 10 times and take the average value as the initial value (before treatment); 3. Hair bundle treatment of sample group (each hair bundle is treated separately): Wet the hair bundle (control the moisture content of the hair bundle to 50% to 65% of the hair weight), apply the test sample evenly on the hair bundle at a dosage of 0.1g / g hair, rub for 1min, rinse with water at a temperature of 37℃ and a flow rate of 4L / min for 1min, and comb the hair every 20 seconds. After rinsing, comb it dry and comb it with a narrow-tooth comb 10 times.

[0046] 4. Treatment of the control group hair bundles (each hair bundle is treated separately): Wet the hair bundles (control the moisture content of the hair bundles to 50% to 65% of the hair weight), apply the test control product evenly on the hair bundles at a dosage of 0.1g / g hair, rub for 1 minute, rinse with water at a temperature of 37℃ and a flow rate of 4L / min for 1 minute, and comb the hair every 20 seconds. After rinsing, comb the hair dry and comb it with a narrow-tooth comb 10 times.

[0047] 5. After the treatment, hang the hair bundle in a constant temperature and humidity chamber at 21±1°C and 50±10%RH for 16-24 hours; 6. Test after using the sample: Use fibra.one to test the combability of the hair bundles. Test each hair bundle 10 times and take the average value as the test value after using the sample (after treatment). Use a combing machine to simulate the scene of people combing their hair. Use a precise force sensor to test the combing force of the hair in the dry state before and after using the sample. The effectiveness of the sample in improving the combability and smoothness of the hair is measured by the significant reduction in the work done by the combing force.

[0048] Hair strand friction work test: 1. Select 10 hair bundles after pretreatment and randomly divide them into 2 groups, 5 bundles in the sample group and 5 bundles in the control group; 2. Initial value test: Use fibra.one to measure the friction work parameter of the hair bundle (pass along the hair scales once for each hair bundle); 3. Hair bundle treatment of sample group (each hair bundle is treated separately): Wet the hair bundle (control the moisture content of the hair bundle to 50% to 65% of the hair weight), apply the test sample evenly on the hair bundle at a dosage of 0.1g / g hair, rub for 1min, rinse with water at a temperature of 37℃ and a flow rate of 4L / min for 1min, and comb the hair every 20 seconds. After rinsing, comb it dry and comb it with a narrow-tooth comb 10 times.

[0049] 4. Treatment of the control group hair bundles (each hair bundle is treated separately): Wet the hair bundles (control the moisture content of the hair bundles to 50% to 65% of the hair weight), apply the test control product evenly on the hair bundles at a dosage of 0.1g / g hair, rub for 1 minute, rinse with water at a temperature of 37℃ and a flow rate of 4L / min for 1 minute, and comb the hair every 20 seconds. After rinsing, comb the hair dry and comb it with a narrow-tooth comb 10 times.

[0050] 5. After the treatment, hang the hair bundle in a constant temperature and humidity chamber at 21±1°C and 50±10%RH for 16-24 hours; 6. Test after using the sample: Use fibra.one to measure the friction work parameter value of the hair strand (pass along the hair scales once for each hair strand).

[0051] Experimental results: Dry combing work difference Figure 1 As shown, the smaller the measured value of dry combing work, the easier it is to comb the hair. The results showed that compared with the control group, the difference in dry combing work in the sample group was significantly reduced (**P<0.01), indicating that the formula with added Yunnan small-grain coffee peptide can significantly improve the dry combing of hair.

[0052] The difference in friction work is Figure 2 As shown, the smaller the measured value of friction work, the easier the hair is to comb. The results showed that compared with the control group, the difference in friction work in the sample group was significantly reduced (*P<0.05), indicating that the formula with added Yunnan small-grain coffee peptide can significantly improve the friction effect of hair.

[0053] Example 3 Anti-inflammatory effect of recombinant human elastin peptide and Yunnan Arabica coffee peptide combination The preparation of recombinant human elastin peptide refers to patent CN118290568A.

[0054] Cytotoxicity Screening Cultured keratinocytes were seeded in a 96-well plate at 0.8-1.0×104 / 100 μL / well, and then placed in an incubator for 18-24 h. The original culture medium was removed, and 100 μL of sample dilutions of different concentrations were added to each well, and exposed in the incubator for 24±0.5 h. 20 μL of thiazolyl blue (MTT) solution was added to each well and incubated at 37°C for 3±0.5 h. Then the MTT solution was removed, and 100 μL of dimethyl sulfoxide (DMSO) was added to each well. After shaking in the dark for 10-15 min, the absorbance value was measured at a wavelength of 570 nm. Anti-inflammatory test Keratinocytes were routinely cultured. Keratinocytes were seeded in 12-well plates at a density of 2.0-3.0×105 / mL / well and returned to the incubator for culture for 18-24 hours. The culture plates were removed and the original culture medium in the wells was discarded. Minimum essential medium (MEM) was added to the negative control (NC), MEM culture medium containing sodium dodecyl sulfate (SLS) was added to the model control group (PC), SLS and MEM culture medium containing different concentrations of test samples were added to each well of the sample group (TA), and SLS and MEM culture medium containing dexamethasone were added to each well of the positive control group (PC). Each group had 3 replicates and was cultured for 24±1 hours. The supernatant was collected and stored at -80℃. Cytokines were determined using ELISA kits.

[0055] The experimental groups are shown in Table 4.

[0056] Table 4

[0057] Experimental results: Effects of different concentrations of LFT02 (weight ratio of 10% Yunnan coffee peptide + 1ppm recombinant human elastin + water to 100%) on the expression of inflammatory factor IL-1α induced by SLS Figure 3 The results showed that both 0.01% LFT02 sample and 0.1% LFT02 sample showed significant inhibitory effects on the inflammatory factor IL-1α.

[0058] Example 4 Bioinformatics predicts the binding of peptide fragments to disulfide bond fragments and predicts the protective effect of products on disulfide bonds.

[0059] Elastin peptide fragments: AAKAGAGLGGVPGVGGLGVSAGAVVPQPGAGVKPGKVPGVGLPGVYPGGVLPGARFPGVGVLPGVPTGAGVKPKAPGVGG (as shown in SEQ ID NO: 2 in the sequence listing) Bioinformatics sequence library analysis: Using bioinformatics to search for hair fragments with high keratin content, keratin sites 140-165 (disulfide bond region): YCMQLSQIQGLIGSVEEQLAQLRCEM (as shown in the sequence list SEQ ID NO: 3) AI predicts protein and short peptide interactions: Alphafold3 is used to predict the Keratin hair fragment and short peptide fragment, and AI and bioinformatics are used to predict the possibility of docking between hair fragments and short peptides. The results are as follows Figure 4 , Figure 5 shown.

[0060] from Figure 4 It can be seen that the above elastin peptide fragment can cover Keratin sites 140-165 (disulfide bond region). from Figure 5 It can be seen that the Arabica coffea peptide KKAGNQGFEYVA (as shown in SEQ ID NO: 1 in the sequence listing) has a certain probability of covering Keratin sites 140-165 (disulfide bond region).

[0061] The above disclosure is only the preferred embodiment of the present invention, which certainly cannot be used to limit the scope of the present invention. Therefore, equivalent changes made according to the claims of the present invention are still within the scope of the present invention.

Claims

1. A composition, characterized in that The invention comprises a recombinant human elastin peptide and a polypeptide derived from Arabica coffee grounds, wherein the elastin peptide and the polypeptide derived from Arabica coffee grounds are combined in a ratio of 1:

100.

2. The composition according to claim 1, characterized in that The polypeptide derived from Arabica coffee grounds includes Yunnan Arabica coffee polypeptide.

3. The composition according to claim 2, characterized in that The preparation of the Yunnan small-grain coffee polypeptide comprises: using Yunnan small-grain coffee grounds as raw materials, reacting with protease, and obtaining the Yunnan small-grain coffee polypeptide.

4. The composition according to claim 3, characterized in that The proteases include ficin.

5. The composition according to claim 2, characterized in that The amino acid sequence of the Yunnan small-grain coffee polypeptide is shown in the sequence listing as SEQ ID NO:

1.

6. A use of the composition according to claim 1, characterized in that: Used for one or more of hair care and scalp care.

7. The use according to claim 6, characterized in that: Used to prepare formulations for improving the combing properties of hair strands.

8. The use according to claim 7, characterized in that: The improving combing performance of the hair bundle includes one or more of improving the dry combing property of the hair bundle and improving the friction property of the hair bundle.

9. The use according to claim 6, characterized in that: Used for preparing anti-inflammatory preparations.

10. The use according to claim 9, characterized in that: Used to prepare preparations for inhibiting the inflammatory factor IL-1α.

11. A preparation obtained according to the use of claim 6.

12. The preparation according to claim 11, characterized in that The preparation comprises one or more of shampoo and hair care essence.

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