Peptide derived from scalp having antibacterial effect on hair loss-inducing harmful bacteria

Abstract

An aspect of the present application relates to a peptide having antibacterial activity against Malassezia, an antibacterial composition including the same, and an antibacterial preparation including the same. The peptide composition having a microorganism group modulating ability of the scalp of the present application shows antibacterial activity against Malassezia, and is helpful to effectively improve the microbial balance of the scalp. Such antibacterial activity can provide a positive effect of enhancing the health of the scalp environment, preventing hair loss, and relieving scalp-related problems.

Description

Technical Field

[0001] One aspect of the present invention relates to a peptide having antibacterial activity against scalp fungi, an antibacterial composition comprising the above-mentioned peptide, and an antibacterial preparation comprising the above-mentioned peptide. Background Technology

[0002] In recent years, with increasing attention to scalp health, research on the scalp microbiome, or scalp microbial ecosystem, has been actively conducted. The scalp microbiome is composed of various bacteria and fungi, which play a crucial role in maintaining the physiological balance of the scalp. A healthy scalp is maintained through a balance between various beneficial and harmful bacteria; however, the excessive proliferation of certain harmful bacteria can lead to various scalp problems such as dandruff, seborrheic dermatitis, and hair loss.

[0003] Some harmful scalp bacteria proliferate in environments with high sebum secretion, inducing inflammatory responses and leading to hair follicle damage. Furthermore, they break down sebum components, generating harmful metabolites that exacerbate scalp inflammation. This imbalance in the scalp microbiome is a major cause of various scalp abnormalities, including hair loss. Therefore, various methods for treating and preventing hair loss are being researched. In addition to existing drug treatments, new formulations of therapeutic agents, protein and antibody drugs, cell therapy agents, and other types of hair loss treatments are being actively developed. In particular, the demand for treatments with fewer side effects and greater patient convenience is increasingly prominent, and the hair loss treatment market is expanding from competition primarily driven by low-priced drugs to a higher-end, more expensive market.

[0004] Existing scalp management products and treatments primarily work by inhibiting microbial growth, but long-term use has limitations such as damaging beneficial bacteria or inducing drug resistance. Therefore, a new approach is needed that can selectively target specific harmful bacteria while maintaining the balance of the scalp microbiome. Summary of the Invention

[0005] Technical issues The purpose of this invention is to provide a peptide with antibacterial activity against scalp fungi, which consists of the amino acid sequence of SEQ ID NO. 1 or SEQ ID NO. 2.

[0006] Furthermore, the object of the present invention is to provide an antibacterial composition comprising the above-mentioned peptide.

[0007] Furthermore, the object of the present invention is to provide an antibacterial preparation comprising the above-mentioned antibacterial composition.

[0008] Solution to the problem To achieve the above objectives, the present invention provides a peptide having antibacterial activity against scalp fungi, which consists of the amino acid sequence of SEQ ID NO. 1 or SEQ ID NO. 2.

[0009] As exemplified by one embodiment of the present invention, the aforementioned scalp fungus may be Malassezia furfur (… Malassezia furfur ), Propionibacterium acnes ( Cutibacterium acnes ) or Staphylococcus epidermidis ( Staphylococcus epidermidis ).

[0010] Furthermore, the present invention provides an antibacterial composition comprising the aforementioned peptide.

[0011] As exemplified by one embodiment of the present invention, the above composition may have a specific effect against Malassezia furfur (a type of Malassezia furfur) originating from the scalp. Malassezia furfur ), Propionibacterium acnes ( Cutibacterium acnes ) or Staphylococcus epidermidis ( Staphylococcus epidermidis ( ) antibacterial activity.

[0012] As in another embodiment of the present invention, the above composition may have the effect of preventing, alleviating or treating hair loss.

[0013] As in another embodiment of the present invention, the aforementioned hair loss can be selected from the group consisting of androgenetic alopecia, alopecia areata, telogen effluvium, traumatic alopecia, trichotillomania, stress alopecia, anagen effluvium, pityriasis alopecia, syphilitic alopecia, seborrheic alopecia, symptomatic alopecia, and congenital alopecia.

[0014] Furthermore, the present invention provides an antibacterial preparation comprising the above-mentioned antibacterial composition.

[0015] As exemplified by one embodiment of the present invention, the above-mentioned formulation can be a topical skin formulation.

[0016] As in another embodiment of the present invention, the above-mentioned topical skin preparations may be selected from the group consisting of creams, gels, ointments, skin emulsions, skin suspensions, transdermal delivery patches, medicated bandages, lotions, hair growth solutions, hair creams, hair lotions, shampoos, conditioners, hair treatments, hair gels, aerosol hair gels, hair pomades, powder gels, hair masks, hair care products, eyebrow growth agents, eyelash growth agents, eyelash nutrients, and combinations thereof.

[0017] As in another embodiment of the invention, the above-described formulation may have a specific effect against Malassezia furfur (a type of Malassezia furfur) originating from the scalp. Malassezia furfur ), Propionibacterium acnes ( Cutibacterium acnes ) or Staphylococcus epidermidis ( Staphylococcus epidermidis ( ) antibacterial activity.

[0018] As in another embodiment of the present invention, the above-mentioned preparation may have the effect of preventing, alleviating or treating hair loss.

[0019] As in another embodiment of the present invention, the aforementioned hair loss can be selected from the group consisting of androgenetic alopecia, alopecia areata, telogen effluvium, traumatic alopecia, trichotillomania, stress alopecia, anagen effluvium, pityriasis alopecia, syphilitic alopecia, seborrheic alopecia, symptomatic alopecia, and congenital alopecia.

[0020] Furthermore, the object of the present invention is to provide a method for antibacterial treatment of scalp bacteria using the above-described composition.

[0021] Furthermore, the present invention aims to provide a method for preventing, alleviating, or treating hair loss using the above-described composition.

[0022] Furthermore, the present invention aims to provide a composition for use in antibacterial purposes against scalp fungi.

[0023] Furthermore, the present invention aims to provide a composition for the prevention, relief or treatment of hair loss.

[0024] The effects of the invention The peptides of this invention, which have scalp microbiome-regulating capabilities, exhibit antibacterial activity against harmful scalp bacteria, helping to effectively improve the scalp's microbial balance. This antibacterial activity can provide positive effects by improving the scalp environment to enhance scalp health, prevent hair loss, and alleviate scalp-related problems.

[0025] In addition, compared with synthetic compound therapeutic agents, peptide therapeutic agents have the advantages of high biocompatibility, high targeting specificity, low risk of side effects, and low possibility of drug resistance.

[0026] The effects of this invention are not limited to those mentioned above, but should be understood to include all effects that can be inferred from the inventive features described in the detailed description or claims of this invention. Attached Figure Description

[0027] Figure 1 To demonstrate based on LC-MS 2 A diagram illustrating the workflow for scalp peptide analysis.

[0028] Figure 2 This diagram illustrates the workflow for discovering scalp-derived peptide candidates with scalp microbiome regulatory capabilities.

[0029] Figure 3 This diagram illustrates a method for verifying the antibacterial activity of peptides using the well diffusion assay.

[0030] Figure 4 A diagram illustrating the microorganisms that make up the scalp microbiome.

[0031] Figure 5 A graph showing the screening results of peptides for antibacterial activity against resident bacteria and harmful bacteria on the scalp.

[0032] Figure 6 A graph showing the results of the determination of the inhibitory concentration (IC50) of peptides against resident bacteria and harmful bacteria on the scalp. Detailed Implementation

[0033] The present invention will now be described in detail.

[0034] To achieve the above objectives, the present invention provides a peptide having antibacterial activity against scalp fungi, which consists of the amino acid sequence of SEQ ID NO. 1 or SEQ ID NO. 2.

[0035] As used in this manual, the term "scalp flora" refers to the sum of all microorganisms found on the scalp. This includes "resident scalp flora," which are normally present on the scalp surface and help maintain a healthy state, as well as "harmful bacteria," which may proliferate excessively under specific conditions such as changes in the scalp environment or a weakened immune system, potentially causing scalp diseases. Scalp flora typically includes various microorganisms such as bacteria, fungi, and yeasts.

[0036] As used in this specification, the term "antimicrobial" refers to the action or activity of inhibiting or eliminating the proliferation of bacteria, fungi, and other microorganisms. Antimicrobial action can include direct bactericidal action and bacteriostatic action that hinders microbial proliferation. Antimicrobial activity is expressed through various chemical or biological mechanisms and can have selectivity or a broad antimicrobial spectrum against specific microorganisms. The peptides described in this specification and antimicrobial compositions including them can prevent or improve hair loss by regulating the microbial balance of the scalp.

[0037] As used in this specification, the term "peptide" refers to a polymer composed of two or more amino acids linked together by peptide bonds.

[0038] The peptides of the present invention can be obtained by various methods known in the art. For example, they can be prepared by using polynucleotide recombination and protein expression systems, or by in vitro synthesis methods such as chemical synthesis of peptides and cell-free protein synthesis, but are not limited to these preparation methods.

[0039] Furthermore, to achieve better chemical stability, enhanced pharmacological properties (half-life, absorption, potency, efficacy, etc.), altered specificity (e.g., broad bioactivity spectrum), and reduced antigenicity, a protecting group may be bound to the N-terminus or C-terminus of the peptide. For example, the protecting group may be acetyl, fluorenyl methoxycarbonyl, formyl, palmitoyl, myristyl, stearoyl, or polyethylene glycol (PEG), but any component capable of modifying the peptide, particularly enhancing its stability, may be included without limitation. The term "stability" refers not only to in vivo stability, which protects the peptide of the present invention from attack by proteolytic enzymes, but also to storage stability (e.g., room temperature storage stability).

[0040] The peptides of the present invention consist of the amino acid sequence of SEQ ID NO. 1 (RKLW) or the amino acid sequence of SEQ ID NO. 2 (RQLW), but are not limited thereto, and can be peptides composed of amino acid sequences having more than 70%, 75%, 80%, 90%, 95%, 96%, 97%, 98% or 99% homology or identity.

[0041] As exemplified by one embodiment of the present invention, the aforementioned scalp fungus may be Malassezia furfur (… Malassezia furfur ), Propionibacterium acnes ( Cutibacterium acnes ) or Staphylococcus epidermidis ( Staphylococcus epidermidis ).

[0042] Among the scalp-derived microorganisms used in this instruction manual, "Malassezia furfur" is mentioned. Malassezia furfur "Propionibacterium acnes" is a lipid-dependent yeast-like fungus found in the sebaceous gland secretions of the scalp and is a major known pathogen associated with dandruff. Cutibacterium acnes "Staphylococcus epidermidis" is a normally resident bacterium found in the sebaceous glands of the skin and scalp, and is a strain known to influence the microbial ecosystem of hair follicles and sebaceous glands. In addition, "Staphylococcus epidermidis" is also present in other bacteria. Staphylococcus epidermidis "It is a Gram-positive coccus that exists as a resident bacterium on the scalp and skin, and can affect the skin barrier function and microbial balance."

[0043] Furthermore, the present invention provides an antibacterial composition comprising the aforementioned peptide.

[0044] As exemplified by one embodiment of the present invention, the above composition may have a specific effect against Malassezia furfur (a type of Malassezia furfur) originating from the scalp. Malassezia furfur ), Propionibacterium acnes ( Cutibacterium acnes) or Staphylococcus epidermidis ( Staphylococcus epidermidis ( ) antibacterial activity.

[0045] As in another embodiment of the present invention, the above composition may have the effect of preventing, alleviating or treating hair loss.

[0046] As used in this specification, the term "hair loss" refers to the absence of hair in areas where hair should normally be present, including a reduction in the amount of hair compared to the normal state. Depending on whether the hair follicles are damaged, the aforementioned hair loss includes both non-scarring and scarring types.

[0047] In this invention, the term "prevention" refers to reducing the risk of developing a disease or disorder, and refers to all behaviors that prevent the progression of one or more clinical symptoms of a disease in an individual exposed to or susceptible to a disease but who has not yet developed the disease or shown symptoms of the disease, thereby inhibiting or delaying the onset of the disease.

[0048] In this invention, the term "relief" refers to the reduction of symptoms of a disease or disorder, including all behaviors that reduce the intensity of symptoms or decrease discomfort by stopping or reducing one or more clinical symptoms of the disease. It refers to all behaviors that improve a patient's quality of life by reducing the frequency, duration, or severity of symptoms, even without eliminating the root cause of the disease.

[0049] In this invention, the term "treatment" refers to alleviating a disease or disorder, including all actions that improve or change the symptoms of a disease in a favorable direction by stopping or reducing the progression of the disease or one or more of its clinical symptoms.

[0050] As in another embodiment of the present invention, the aforementioned hair loss can be selected from the group consisting of androgenetic alopecia, alopecia areata, telogen effluvium, traumatic alopecia, trichotillomania, stress alopecia, anagen effluvium, pityriasis alopecia, syphilitic alopecia, seborrheic alopecia, symptomatic alopecia, and congenital alopecia.

[0051] Furthermore, the present invention provides an antibacterial preparation comprising the above-mentioned antibacterial composition.

[0052] As used in this instruction manual, the term "preparation" refers to a formulation made by mixing one or more active ingredients with appropriate pharmaceutical or chemical components to achieve a specific functional purpose. Preparations include various types based on their physical form or mode of application. Preparations can be provided in physical states such as solid, liquid, and semi-solid, and are classified as oral, injectable, or for direct external application based on their intended use and site of application.

[0053] As exemplified by one embodiment of the present invention, the above-mentioned formulation can be a topical skin formulation.

[0054] In particular, as used in this specification, "topical skin preparation" refers to a preparation that provides specific functional effects by being applied to human skin. Such topical skin preparations are used for antibacterial, moisturizing, anti-inflammatory, or other therapeutic and cosmetic purposes, and can help maintain or improve scalp health by including compositions of the present invention.

[0055] As in another embodiment of the present invention, the above-mentioned topical skin preparations may be selected from the group consisting of creams, gels, ointments, skin emulsions, skin suspensions, transdermal delivery patches, medicated bandages, lotions, hair growth solutions, hair creams, hair lotions, shampoos, conditioners, hair treatments, hair gels, aerosol hair gels, hair pomades, powder gels, hair masks, hair care products, eyebrow growth agents, eyelash growth agents, eyelash nutrients, and combinations thereof.

[0056] As in another embodiment of the invention, the above-described formulation may have a specific effect against Malassezia furfur (a type of Malassezia furfur) originating from the scalp. Malassezia furfur ), Propionibacterium acnes ( Cutibacterium acnes ) or Staphylococcus epidermidis ( Staphylococcus epidermidis ( ) antibacterial activity.

[0057] As in another embodiment of the present invention, the above-mentioned preparation may have the effect of preventing, alleviating or treating hair loss.

[0058] As in another embodiment of the present invention, the aforementioned hair loss can be selected from the group consisting of androgenetic alopecia, alopecia areata, telogen effluvium, traumatic alopecia, trichotillomania, stress alopecia, anagen effluvium, pityriasis alopecia, syphilitic alopecia, seborrheic alopecia, symptomatic alopecia, and congenital alopecia.

[0059] In the case that the antibacterial composition of the present invention is a pharmaceutical composition, it can be used against Malassezia furfur (a type of Malassezia) originating from the scalp. Malassezia furfur), Propionibacterium acnes ( Cutibacterium acnes ) or Staphylococcus epidermidis ( Staphylococcus epidermidis The pharmaceutical composition of this invention is intended for antibacterial use. It may contain the aforementioned peptides as active ingredients, and may also contain pharmaceutically permissible carriers. These pharmaceutically permissible carriers are those conventionally used in formulation and include, but are not limited to, physiological saline, sterile water, Ringer's solution, buffered physiological saline, cyclodextrin, glucose solution, maltodextrin solution, glycerol, ethanol, liposomes, etc., and may also include antioxidants, buffer solutions, and other conventional additives as needed. Furthermore, diluents, dispersants, surfactants, binders, and lubricants may be added to formulate it into injections (e.g., aqueous solutions, suspensions, emulsions, etc.), pills, capsules, granules, or tablets. For suitable pharmaceutically permissible carriers and formulation, refer to the methods disclosed in Remington's literature, and formulate appropriately according to each component. The pharmaceutical composition of this invention is not particularly limited to a specific dosage form and may be formulated as an injection, inhalation, topical preparation, or oral preparation.

[0060] The pharmaceutical compositions of the present invention can be administered orally or parenterally according to the desired method (e.g., suitable for intravenous, subcutaneous, skin, nasal, and respiratory tract administration). The dosage varies depending on the patient's condition and weight, disease severity, drug form, route of administration, and time of administration, and can be appropriately selected by those skilled in the art.

[0061] The compositions of the present invention are administered in pharmaceutically effective amounts. In this invention, a "pharmaceutically effective amount" refers to an amount sufficient to treat a disease with a reasonable benefit / risk ratio suitable for medical treatment. The effective dose level can be determined based on the patient's disease type, disease severity, drug activity, drug sensitivity, timing of administration, route of administration, excretion rate, treatment duration, concurrent medications, and other factors well-known in the medical field. The compositions of the present invention can be administered as a single therapeutic agent or in combination with other therapeutic agents, sequentially or simultaneously with conventional therapeutic agents, and in single or multiple doses. Considering all the foregoing factors, it is important to administer the minimum amount necessary to achieve maximum effect without any side effects, and this can be readily determined by those skilled in the art.

[0062] In the case where the antibacterial composition of the present invention is a cosmetic composition, it can be used against Malassezia furfur (a type of bacteria) originating from the scalp. Malassezia furfur ), Propionibacterium acnes ( Cutibacterium acnes ) or Staphylococcus epidermidis ( Staphylococcus epidermidis The above-mentioned cosmetic composition may contain a cosmetically effective amount of the above-mentioned peptide and a cosmetically permissible carrier, wherein the cosmetically effective amount refers to an amount sufficient to achieve the aforementioned anti-hair loss or hair growth promotion effects.

[0063] The cosmetic compositions of the present invention may contain conventionally used adjuvants, such as hydrophilic or lipophilic gelling agents, hydrophilic or lipophilic active agents, preservatives, antioxidants, solvents, fragrances, fillers, blocking agents, pigments, deodorants, and dyes.

[0064] In one embodiment, the cosmetic composition of the present invention can be a topical skin preparation dosage form, which can be selected from the group consisting of hair growth lotion, hair cream, hair lotion, shampoo, hair conditioner, hair conditioner, hair gel, aerosol hair gel, hair balm, powder gel, hair mask, hair care agent, eyebrow growth agent, eyelash growth agent and eyelash nutrient agent.

[0065] In the case where the antibacterial composition of the present invention is a food composition, it can be used against Malassezia furfur from the scalp ( Malassezia furfur ), Propionibacterium acnes ( Cutibacterium acnes ) or Staphylococcus epidermidis ( Staphylococcus epidermidis The antibacterial uses of the present invention are described. The food compositions of the present invention include all forms such as functional foods, nutritional supplements, health foods, food additives, and animal feed, intended for consumption by animals, including humans and livestock. The above-mentioned types of food compositions can be prepared in various forms according to conventional methods known in the art.

[0066] The food compositions of the above types can be prepared in various forms according to conventional methods known in the art. As general foods, for example, they can be contained in beverages (including alcoholic beverages), fruits and their processed products (e.g., canned fruits, bottled and canned goods, jams, marmalade, etc.), fish, meat and their processed products (e.g., ham, sausages, corned beef, etc.), breads and noodles (e.g., udon noodles, soba noodles, ramen, pasta, macaroni, etc.), fruit juices, various beverages, biscuits, sugar, dairy products (e.g., butter, cheese, etc.), edible vegetable oils, margarine, plant-based proteins, soft canned foods, frozen foods, and various seasonings (e.g., miso, soy sauce, sauces, etc.), wherein the above-mentioned compositions contain the above-mentioned peptides as active ingredients.

[0067] The present invention will be described in more detail below through embodiments to facilitate understanding. However, the following embodiments are merely illustrative of the invention, and the scope of the invention is not limited to the following embodiments. The embodiments of the present invention are provided to provide a more complete explanation of the invention to those skilled in the art.

[0068] Example Example 1. Computer simulation of scalp-derived peptides with scalp microbiome regulatory capabilities ( in silico )explore Based on LC-MS 2 The workflow for scalp peptide analysis is as follows: Figure 1 First, tissue samples were collected from the scalp of both the non-hair loss group and the hair loss group; this was known as scalp surface tissue biopsy. The samples from the non-hair loss group and the hair loss group were stored in separate containers to prevent mixing. The collected samples underwent a concentration and desalting process, during which impurities were removed and peptide components were concentrated to make them readily analyzeable.

[0069] Then, via LC-MS 2 The analyzer analyzes the sample and acquires MS spectrum data. MS spectra measure the mass-to-charge ratio (m / z) of peptides in the sample, providing insights into the peptide's structure and properties. The acquired MS spectrum data is then subjected to de novo sequencing (…). De novo The sequencing process involves directly interpreting the amino acid sequence of a peptide from an MS spectrum. This is used to construct a scalp peptide library.

[0070] Based on the constructed peptide library, computer simulations were performed ( in silico The process involves analyzing and predicting the functionality of each peptide. During this process, an AI-based predictive model is used to assess the potential physiological activity of each peptide, with a particular focus on peptides capable of regulating the scalp microbiome. Based on the prediction results, bioactive peptides are screened. Finally, the screened functional peptides undergo direct activity verification and screening in the laboratory, thereby identifying effective scalp-derived peptides that can practically regulate the scalp microbiome.

[0071] The discovery process for scalp-derived peptide candidates with scalp microbiome regulation capabilities is carried out through the following steps ( Figure 2 ).

[0072] In the initial analysis step, de novo sequencing is performed. De novo Sequencing yielded a total of 198,384 peptides. During this process, strict baseline values ​​specified by each analytical software were applied; PEAKS software only screened de novo scoring peptides. De novo For peptides with a score exceeding 55, Novor software only filters those with a de novo score. De novo Peptides with a score exceeding 50 were included. This quality management ensured the reliability of 16,475 peptide sequences.

[0073] In subsequent steps, an in-house prediction model called pQSAR v0.1 was used to screen for anti-inflammatory peptides (AIPs). Only peptides with an AIP score greater than 0.5 were selected, excluding 14,694 non-AIPs in the process, ensuring that 1,781 scalp-derived functional peptides with anti-inflammatory activity were predicted.

[0074] In the final experimental data analysis step, the statistical significance between the non-hair loss group (NA) and the hair loss group (AA) was analyzed. A p-value less than 0.05 was considered statistically significant. After excluding 1778 peptides with no significant difference, two peptides showing significant differences were finally selected: RKLW (Arg-Lys-Leu-Trp; SEQ ID NO. 1) and RQLW (Arg-Gln-Leu-Trp; SEQ ID NO. 2).

[0075] The two peptides selected in this way were classified as potential bioactive peptides (BPs) for preventing hair loss (anti-alopecia), and they are expected to effectively regulate the scalp microbiome. Through this systematic and phased screening process, two peptides, RKLW (Arg-Lys-Leu-Trp) and RQLW (Arg-Gln-Leu-Trp), were identified from 198,384 initial peptides, which are predicted to have actual preventive or alleviating effects on hair loss.

[0076] Example 2. Validation of peptides that regulate the scalp microbiome in vitro ( in vitro )effect The antimicrobial activity of the two discovered peptides was verified using the well diffusion assay. The well diffusion assay is a standardized experimental method for quantitatively assessing the antimicrobial activity of peptides, and the strength of antimicrobial activity can be objectively determined by measuring the size of the inhibition zone.

[0077] The process for verifying the antibacterial activity of peptides using the well diffusion assay is as follows ( Figure 3 First, prepare hard agar (1.5%) and soft agar (0.75%) separately. Pour the hard agar into a petri dish to form a basal culture medium layer. Then, mix the soft agar with the target microorganism and overlay it on top of the hard agar. Make wells in the resulting bilayer culture medium and inject the peptide sample to be validated or the control group into each well.

[0078] After incubation, observe the zone of inhibition that forms around each well. If an inhibition zone forms, it means that there is an antimicrobial effect. If no inhibition zone forms, it is determined to be ineffective.

[0079] Referencing the composition of the scalp microbiome (microorganisms) Figure 4 , Suzuki et al Microorganisms 2021, 9(10), 2132), selected the main microorganisms of the scalp microbiome—two bacteria (Staphylococcus epidermidis (S. epidermidis)). Staphylococcus epidermidis ), Propionibacterium acnes ( Cutibacterium acnes )) and a fungus (Malassezia furfur) Malassezia furfur ()) were used as screening targets for antibacterial activity (Table 1).

[0080] [Table 1] Scalp Microbiome Information

[0081] Malassezia (a type of microorganism found on the scalp) Malassezia Malassezia is a lipid-dependent yeast found on the scalp, possessing a variety of genes for lipases that break down sebum. This is because Malassezia lacks genes for fatty acid synthesis and must rely on external lipid sources. Malassezia has 14 lipase and 9 phospholipase genes, most of which are predicted to be secreted enzymes. These enzymes break down lipids in the scalp to produce fatty acids, which can trigger skin irritation and inflammation. Furthermore, it promotes excessive keratinization, leading to dandruff, and may be accompanied by scalp itching and redness.

[0082] Staphylococcus spp. Staphylococcus ) and dermatobacteria ( Cutibacterium Scalp microbiota (SCM) is the main genus of microorganisms in the scalp. Under normal conditions, it plays an important role in maintaining scalp pH and preventing the invasion of other harmful bacteria, which are essential for skin health. However, if changes in the scalp environment disrupt the balance of these microorganisms, various problems can arise.

[0083] Malassezia furfur ( Malassezia furfur Yeast (Vulgodiospora) is a common yeast found on the scalp. It is a major pathogen of dandruff and seborrheic dermatitis. It normally resides on the scalp but can proliferate excessively due to excessive sebum secretion or environmental factors such as humidity. It produces fatty acids by breaking down lipids in the skin, which can cause skin irritation and inflammation. It causes dandruff by promoting excessive production of the stratum corneum, which may be accompanied by itching and redness of the scalp.

[0084] Staphylococcus epidermidis ( Staphylococcus epidermidisPropionibacterium acnes is a normal resident flora of the skin and scalp, usually non-pathogenic. It plays an important role in maintaining skin health, such as maintaining skin pH and preventing the invasion of other harmful bacteria. However, it can trigger opportunistic infections when immunity is weakened or the skin barrier is compromised. Due to its ability to form biofilms, it is also one of the main pathogens of medical device-related infections. On the scalp, it interacts with Propionibacterium acnes to help maintain skin homeostasis, but sometimes it can invade sebaceous glands and cause inflammation.

[0085] Propionibacterium acnes ( Cutibacterium acnes These are common, resident bacteria found in the sebaceous glands of the skin and scalp, playing an important role in skin health. They are usually non-pathogenic, but can trigger inflammatory skin conditions such as acne when there is excessive sebum production or a weakened immune system. They are also known to be found on the scalp as strains that influence the microbial ecosystem of hair follicles and sebaceous glands.

[0086] The aim was to verify the antibacterial effect of the peptide against the three types of harmful and resident bacteria on the scalp mentioned above. Figure 5 The selected bacteria that causes scalp inflammation is Propionibacterium acnes (…). Cutibacterium acnes ATCC 6919, Malassezia furfur, a fungus that causes dandruff and seborrheic dermatitis. Malassezia furfur ATCC 14521T and Staphylococcus epidermidis, a normal resident bacterium of the scalp ( Staphylococcus epidermidis ATCC 12228 was used as the experimental subject. The antibacterial activity of two peptides, RKLW (Arg-Lys-Leu-Trp) and RQLW (Arg-Gln-Leu-Trp), was evaluated for these strains.

[0087] The antibacterial activity of each peptide was determined using a well diffusion assay. The peptides were diluted and used at four concentrations: 50 mg / ml, 25 mg / ml, 12.5 mg / ml, and 6.25 mg / ml. To ensure the reliability of the experiment, an appropriate control group was established. This assay was targeted at the fungus *Malassezia furfur* (…). M. furfur Fluconazole (0.5 mg / ml) was used as a positive control group to target the bacteria Staphylococcus epidermidis (Staphylococcus epidermidis). S. epidermidis ) and Propionibacterium acnes ( C. acnes Nisin (2 mg / ml) was used as a positive control, and distilled water was used as a negative control. Six wells were formed in each culture medium, and each concentration of peptide and the control group were treated separately.

[0088] RKLW peptide against Malassezia furfur ( M. furfur It exhibits the strongest antibacterial activity, which is in Figure 5 It is identified by the distinct transparent ring (marked by a yellow arrow) that forms around the pore. For Staphylococcus epidermidis (… S. epidermidisWeak antibacterial activity was also observed (yellow arrow), but compared to Malassezia furfur ( M. furfur The size of the inhibition zone is relatively small.

[0089] However, at the same concentration, the RQLW peptides did not show significant antibacterial activity against any of the three strains. In particular, both peptides showed no activity against *Propionibacterium acnes* (…). C. acnes No inhibition ring was formed around the hole.

[0090] Moreover, for Malassezia furfur ( M. furfur ATCC 14521T, Staphylococcus epidermidis ( S. epidermidis ATCC 12228 and Propionibacterium acnes ( C. acnes According to ATCC 6919, the antibacterial activity of RKLW and RQLW peptides was evaluated by a broth microdilution assay. Figure 6 Each graph represents the peptide concentration used (log). 10 The antibacterial activity of the [compound] (μg / μL) and its corresponding reaction ratio (%) was analyzed by comparing IC50 values. The results confirmed that not only RKLW peptide, but also RQLW peptide showed activity against Malassezia furfur (…). M. furfur RKLW showed antibacterial activity against Staphylococcus epidermidis (Staphylococcus epidermidis). Furthermore, RKLW demonstrated antibacterial activity against Staphylococcus epidermidis (Staphylococcus epidermidis). S. epidermidis ) and Propionibacterium acnes ( C. acnes The antibacterial activity of the sample was weak, and the IC50 values ​​showed a tendency to be lower in RKLW than in RQLW.

[0091] That is, it was confirmed that RKLW has higher overall antibacterial activity against all three strains compared to RQLW, especially showing strong inhibitory effect with lower IC50 values ​​against strains that cause scalp inflammation.

[0092] Therefore, RKLW peptide (SEQ ID NO. 1) and RQLW peptide (SEQ ID NO. 2) were selected as the final candidate substances. These peptides are effective against the harmful scalp bacteria Malassezia furfur (Malassezia furfur). M. furfur ), Propionibacterium acnes ( C. acnes ) or Staphylococcus epidermidis, a resident bacterium of the scalp ( S. epidermidis This demonstrates selective antimicrobial properties, exhibiting antimicrobial activity. This selective antimicrobial spectrum suggests the potential of it as a therapeutic agent that can target specific pathogenic strains without significantly disrupting the normal microbial balance of the scalp.

[0093] The foregoing description of the present invention is illustrative, and those skilled in the art will understand that it can be easily modified into other specific forms without changing the technical concept or essential features of the invention. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not limiting.

Claims

1. A peptide having antimicrobial activity against scalp fungi, comprising the amino acid sequence of SEQ ID NO. 1 or SEQ ID NO.

2.

2. The peptide according to claim 1, wherein, The aforementioned scalp bacteria are Malassezia furfur, Propionibacterium acnes, or Staphylococcus epidermidis.

3. An antibacterial composition comprising the peptide according to any one of claims 1 or 2.

4. The antibacterial composition according to claim 3, wherein, The above composition has antibacterial activity against Malassezia furfur, Propionibacterium acnes, or Staphylococcus epidermidis originating from the scalp.

5. The antibacterial composition according to claim 3, wherein, The above composition has the effect of preventing, alleviating or treating hair loss.

6. The antibacterial composition according to claim 5, wherein, The hair loss mentioned above was selected from the group consisting of androgenetic alopecia, alopecia areata, telogen effluvium, traumatic alopecia, trichotillomania, stress-related alopecia, anagen effluvium, pityriasis alopecia, syphilitic alopecia, seborrheic alopecia, symptomatic alopecia, and congenital alopecia.

7. An antibacterial preparation comprising the antibacterial composition of claim 3.

8. The antibacterial agent according to claim 7, wherein, The above preparations are for external use on the skin.

9. The antibacterial agent according to claim 8, wherein, The above-mentioned topical skin preparations are selected from the group consisting of creams, gels, ointments, skin emulsions, skin suspensions, transdermal delivery patches, medicated bandages, lotions, hair growth tonics, hair creams, hair lotions, shampoos, conditioners, hair treatments, hair gels, aerosol hair gels, hair pomades, powder gels, hair masks, hair care products, eyebrow growth agents, eyelash growth agents, eyelash nutrients, and combinations thereof.

10. The antibacterial agent according to claim 7, wherein, It has antibacterial activity against Malassezia furfur, Propionibacterium acnes, or Staphylococcus epidermidis originating from the scalp.

11. The antibacterial agent according to claim 7, wherein, It has the effect of preventing, alleviating or treating hair loss.

12. The antibacterial agent according to claim 11, wherein, The hair loss mentioned above was selected from the group consisting of androgenetic alopecia, alopecia areata, telogen effluvium, traumatic alopecia, trichotillomania, stress-related alopecia, anagen effluvium, pityriasis alopecia, syphilitic alopecia, seborrheic alopecia, symptomatic alopecia, and congenital alopecia.

Images