Plant active peptide and its use in products for preventing or treating acne

Abstract

The application discloses a kind of plant active peptide and its application in the product for preventing or treating acne, it relates to the technical field of plant extract, the amino acid sequence of the plant active peptide is Pro-Gly-Lys-Glu (PGKE). In vitro antibacterial experiment shows that the plant active peptide has significant inhibitory effect on propionibacterium acnes, acne mouse model experiment confirms that after the external use of 0.5% plant active peptide PGKE-containing gel is administered, it can quickly reduce the volume of acne nodule, reduce redness and inflammation.The plant active peptide of the application is natural, high safety, simple preparation process, and can be used for preparing the product for preventing or treating acne, such as drug, medical device or cosmetic, and has wide clinical application and market development prospect.

Description

Technical Field

[0001] This invention belongs to the technical field of plant extracts, and particularly relates to a plant active peptide and its application in products for the prevention or treatment of acne. Background Technology

[0002] Acne is a common chronic inflammatory skin disease that frequently occurs in areas rich in sebaceous glands, such as the face, chest, and back. It mainly manifests as comedones, papules, pustules, nodules, and cysts. In severe cases, it can lead to permanent scarring and pigmentation, significantly impacting the physical and mental health of patients. Statistics show that approximately 85% of adolescents and young adults worldwide have experienced acne, and the incidence of adult acne has been on the rise in recent years.

[0003] The pathogenesis of acne is complex, mainly involving four aspects: excessive sebum secretion, abnormal keratinization of the hair follicle opening, excessive proliferation of Propionibacterium acnes, and inflammatory immune response. Among these, Propionibacterium acnes is considered one of the key factors inducing and aggravating acne inflammation. This bacterium breaks down triglycerides in sebum to produce free fatty acids, stimulating an inflammatory response in the hair follicle and surrounding tissues. Simultaneously, its own metabolites can also activate immune pathways such as Toll-like receptors, further exacerbating inflammation.

[0004] Currently, the commonly used anti-acne drugs in clinical practice mainly include the following categories: (1) Retinoic acid drugs, such as retinoic acid cream and adapalene gel, which work by regulating follicular keratinization and inhibiting inflammation, but often cause local adverse reactions such as dry skin, peeling, and stinging; (2) Antibiotic drugs, such as clindamycin and erythromycin, which improve symptoms by inhibiting the growth of Propionibacterium acnes, but long-term use has led to increasingly prominent problems of bacterial resistance; (3) Benzoyl peroxide, which has bactericidal and mild keratolytic effects, but some patients may experience allergic reactions and skin irritation; (4) Oral isotretinoin, which has a definite curative effect but many side effects, such as teratogenicity, liver damage, and dyslipidemia, which limit its widespread application.

[0005] Given the problems of drug resistance, local irritation, and systemic side effects associated with existing anti-acne drugs, developing novel anti-acne active substances that are naturally derived, highly safe, and have a clearly defined mechanism of action is of significant clinical importance and application value. Plant-derived bioactive peptides have received widespread attention in the fields of skin care and functional cosmetics in recent years due to their advantages such as small molecular weight, easy absorption by the skin, diverse biological activities, and low toxicity. Soy protein is abundant and inexpensive, and its enzymatic hydrolysis products contain a variety of bioactive small molecule peptides; however, there is currently a lack of research on its application in the prevention and treatment of acne.

[0006] Therefore, there is an urgent need to develop a plant-based bioactive peptide derived from soybean protein that has significant anti-acne activity and high safety to meet the growing demand for acne prevention and treatment. Summary of the Invention

[0007] In a first aspect, the present invention provides a plant bioactive peptide, wherein the amino acid sequence of the plant bioactive peptide is Pro-Gly-Lys-Glu.

[0008] In a second aspect, the present invention provides a composition comprising the plant-active peptides described in the first aspect.

[0009] Optionally, the composition is a topical preparation selected from at least one of gels, creams, ointments, lotions, sprays, or patches.

[0010] Optionally, the composition is a gelling agent, and the plant active peptides in the gelling agent have a mass percentage of 0.1% to 5%.

[0011] Thirdly, the present invention provides the use of the plant-based active peptides described in the first aspect or the compositions described in the second aspect in the preparation of products for the prevention or treatment of acne.

[0012] Optionally, the product may be a pharmaceutical, medical device, or cosmetic.

[0013] In summary, the present invention has the following advantages compared with the prior art: 1. Natural source and high safety: The plant active peptides of this invention are derived from soybean protein and obtained through enzymatic hydrolysis, ultrafiltration separation and liquid chromatography-mass spectrometry identification. The raw materials are widely available and inexpensive. Moreover, the peptides themselves are natural protein degradation products, with good biocompatibility and high safety, making them suitable for long-term use in skin care. 2. Significant anti-acne activity: In vitro antibacterial experiments showed that the minimum inhibitory concentration (MIC) of the plant active peptide PGKE of this invention against Propionibacterium acnes was only 25 μM, and it also had varying degrees of inhibitory effects on other common skin pathogens such as Staphylococcus aureus, showing good selectivity and broad-spectrum antibacterial potential. 3. Excellent in vivo therapeutic effect: Acne mouse model experiments have shown that after 4 days of topical application of gel containing 0.5% plant active peptide PGKE, the volume of acne nodules is reduced by about 80%, which is significantly better than commercially available 0.1% adapalene gel, and has a faster onset of action and more thorough reduction of redness and inflammation. Attached Figure Description

[0014] Figure 1 This is a graph showing the volume change of the plant-based bioactive peptides of this invention in a mouse model of acne treatment. Detailed Implementation

[0015] The present invention will be further described in detail below with reference to specific embodiments. These embodiments are only used to explain the present invention and do not constitute a limitation on the scope of protection of the present invention. Experimental methods not specified in these embodiments can be performed with reference to conventional production processes; unless otherwise specified, the reagents used are commercially available or obtainable through public channels.

[0016] Example 1: Soybean protein hydrolysate

[0017] 125g of soybean protein powder was mixed with deionized water to prepare a 1000mL solution, and extracted by stirring in a 55℃ water bath for 1 hour. First, 0.2kg of alkaline protease was added to adjust the pH to 8.0, and enzymatic hydrolysis was carried out at 55℃ for 5 hours, followed by enzyme inactivation at 85℃ for 15 minutes. Then, the temperature was lowered to 55℃, 0.15kg of papain was added, and the pH was adjusted to 6.0. Enzymatic hydrolysis was carried out at 55℃ for 3 hours, followed by enzyme inactivation at 85℃ for 15 minutes. The hydrolysate was centrifuged at 4℃ and 10000 r / min for 30 minutes, and the supernatant was collected. The supernatant was then filtered through 0.45 μm and 0.22 μm microfiltration membranes for sterilization and impurity removal. Subsequently, the filtrate was separated using an ultrafiltration membrane module with a molecular weight cutoff of 3 kDa, and the permeate with a molecular weight <3 kDa was collected to obtain the soybean protein hydrolysate.

[0018] Example 2 Plant bioactive peptides

[0019] The soybean protein hydrolysate from Example 1 was analyzed by LC-MS / MS.

[0020] The mass spectrometry data were analyzed using the built-in analysis functions of the BioChemia (https: / / biochemia.uwm.edu.pl / biopep / peptide_data.php) and InterPro (https: / / www.ebi.ac.uk / interpro / ) databases to confirm the potential biological activities of the candidate sequences, and to fully annotate their potential biological activities. Finally, the plant bioactive peptides with the amino acid sequence PGKE were identified and screened from the enzymatic digest.

[0021] Synthesis of target peptide: Based on the identification and screening results, the above-mentioned plant bioactive peptide PGKE was synthesized in solid phase and purified by preparative high performance liquid chromatography to obtain high-purity (>95%) peptide standards.

[0022] Example 3 Antibacterial assay of plant bioactive peptides

[0023] Bacterial culture: Propionibacterium acnes (ATCC6919) was grown anaerobically in BHI medium; Staphylococcus aureus (ATCC29213), Candida albicans (ATCC10231), Bacillus subtilis (ATCC63501), and Escherichia coli (ATCC25922) were grown aerobically in agar medium. All strains were cultured at 37°C.

[0024] Preparation of bacterial suspension: The concentration of bacteria was determined using a McFarland turbidity tube. The turbidity was adjusted to 0.5 McFarland turbidity, at which point the bacterial colony count was approximately 1 × 10⁻⁶. 8 The concentration was CFU / mL, and then diluted to a bacterial suspension of 10⁵-10⁶ CFU / mL.

[0025] Antibacterial test: The concentration of the polypeptide PGKE was 0-800 μM. During the logarithmic growth phase, bacterial suspensions were taken, and *Propionibacterium acnes* (ATCC6919) was diluted to a final concentration of 1×10⁻⁶. 6 CFU / mL, other strains diluted to 1×10 5 CFU / mL. Add 100 μL of bacterial suspension to each well. After static incubation of *Propionibacterium acnes* for 96 hours and other strains for 17-20 hours, measure the absorbance of the bacterial suspension at 600 nm using a microplate reader. The MIC value is defined as the lowest drug concentration at which no bacterial growth is observed. Repeat the test 4 to 6 times independently.

[0026] The results are shown in Table 1. The antimicrobial activity of plant bioactive peptides against Propionibacterium acnes was detected by MIC. The MIC of plant bioactive peptide PGKE against Propionibacterium acnes was 25 μM, which was lower than that of other bacterial strains tested.

[0027] Table 1

[0028] Example 4: Anti-acne efficacy of plant-based bioactive peptides in an acne mouse model

[0029] BALB / c mice (3-5 weeks old) were used as the model animals. Hair was removed from the backs of the mice using an animal shaver, and depilatory cream was applied. The model was established 48 hours after hair removal. Each mouse was intradermally injected with 25 μL of Propionibacterium acnes PBS buffer solution (1×10⁹ CFU / mL) on its back. Approximately two days later, noticeable swelling and redness were observed at the injection site. The mice were then randomly assigned to groups and administered the medication.

[0030] Twelve BALB / c mice were randomly divided into three groups (n=4): a model group, a control group (adapalene gel), and an experimental group (0.5% plant bioactive peptide PGKE gel). The control group received 50 mg of adapalene gel topically applied to the back of each BALB / c mouse, while the experimental group received 50 mg of gel containing 0.5% plant bioactive peptide PGKE topically applied to the back of each BALB / c mouse (except for the active ingredient, it was identical to the adapalene gel). The model group received no treatment. The volume of the skin bulge was measured daily during the experiment. At the end of the experiment, the mice were euthanized by cervical dislocation.

[0031] The results are as follows Figure 1 As shown, the plant-based active peptide PGKE of this invention can treat acne. After 4 days of administration, it can suppress acne to a smaller level and reduce redness and inflammation, while the acne volume in the non-administered group did not change significantly.

[0032] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A plant-based bioactive peptide, characterized in that, The amino acid sequence of the plant bioactive peptide is Pro-Gly-Lys-Glu.

2. A composition, characterized in that, It contains the plant-active peptides as described in claim 1.

3. The composition according to claim 2, characterized in that, The composition is a topical preparation selected from at least one of gels, creams, ointments, lotions, sprays, or patches.

4. The composition according to claim 3, characterized in that, The composition is a gelling agent, and the plant active peptides in the gelling agent have a mass percentage content of 0.1% to 5%.

5. The use of the plant-based active peptide of claim 1 or the composition of any one of claims 2-4 in the preparation of products for the prevention or treatment of acne.

6. The application according to claim 5, characterized in that, The product in question is a pharmaceutical, medical device, or cosmetic.

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