Anti-alopecia polypeptide and related substance and application thereof

Abstract

The invention discloses an anti-alopecia and anti-dermatitis polypeptide and related substances and application thereof, and particularly discloses a polypeptide sequence shown as any one of SEQ ID NO.1-7, a polypeptide containing the sequence, and a preparation method of the anti-alopecia and anti-dermatitis polypeptide. The homology of the sequences is higher than 50%, and the derivatives of the sequences are used for preparing medicines for treating androgen / seborrheic hair follicle decline and / or alopecia, hair follicle decline and / or alopecia caused by medicines, seborrheic acne and / or dermatitis, and acne and / or dermatitis caused by medicines; the invention also discloses application of the polypeptide and the derivative thereof in preparation of medicines, medical and beauty supplies, cosmetics, health care products, food and additives for resisting, preventing and / or treating hair follicle decline, alopecia, acne and dermatitis.

Description

Technical Field

[0001] The present invention belongs to the field of medical biology and relates to a class of polypeptides and related substances for combating problems such as hair loss and their applications. Background Art

[0002] Hair loss (also known as hair loss) refers to the phenomenon of hair loss, which can be categorized as physiological or pathological. Physiological hair loss refers to the normal shedding of hair during the catagen and telogen phases to maintain a normal hair count. Pathological hair loss, on the other hand, refers to abnormal or excessive hair loss.

[0003] There are many reasons for hair loss, including genetic factors, immune factors, excessive mental stress, staying up late, drugs (such as hair loss caused by chemotherapy), environmental factors, endocrine disorders, etc.

[0004] According to the cause, hair loss can be divided into stress-induced alopecia, androgenic (seborrheic) alopecia, pathological alopecia, physical / chemical alopecia, nutritional alopecia, obesity-related alopecia, hereditary alopecia, etc. Currently, there are two drugs approved by the U.S. Food and Drug Administration (FDA) for the treatment of hair loss: minoxidil and finasteride. Minoxidil is a topical drug applied to the scalp, which has a "rapid shedding period", takes a long time to take effect, and has certain side effects; finasteride is an oral drug, but it has a major hidden danger to female fertility and is ineffective for postmenopausal women, so it is not recommended for women. Clinically, there is a need to develop safer, more effective, and suitable intervention drugs for a variety of populations and types of hair loss.

[0005] Androgenic alopecia (AGA) is divided into male pattern (MPHL) and female pattern (FPHL) and is caused by genetic factors and the effects of androgens. The differences in sex hormones between men and women lead to significant differences in the causes, diagnosis, and treatment of male and female hair loss. Men have higher levels of androgen. Once testosterone reaches the hair follicles, it is converted to dihydrotestosterone (DHT) by 5α-reductase, causing hair follicle atrophy and gradual thinning of the hair. In men with genetic predispositions, hair loss can manifest as a receding hairline, M-shaped hair loss, and ultimately, baldness. In women, androgen levels are lower, and estrogen can suppress androgen production. However, when hormonal balance is disrupted or estrogen levels decrease after middle age, hair loss due to insufficient estrogen secretion and excessive androgen production can occur. This typically presents as progressive hair loss, characterized by progressive follicle miniaturization and thinning of the hair at the center of the head.

[0006] The role of androgens in the pathogenesis of male-pattern androgenic alopecia has been basically clarified, while the mechanism of action in the pathogenesis of female-pattern androgenic alopecia is still unclear. In addition to androgens, non-androgen-dependent pathways may also be involved, such as chronic low-grade inflammation of the scalp, androgen insensitivity syndrome, estrogen deficiency, abnormal thyroid function, metabolic syndrome, etc. [Referenced from "Chinese Expert Consensus on the Diagnosis and Treatment of Female Androgenic Alopecia (2022 Edition)"].

[0007] Drug-induced alopecia occurs when drugs, taken orally or injected, damage the proliferating hair follicle stem cells, leading to atrophy of the follicles and brittle hair, which can break or fall out easily, causing hair loss. Common drugs that cause alopecia include anticancer drugs, anti-tumor drugs, chemical agents, and oral contraceptives.

[0008] Irisin is a hormone secreted by skeletal muscle. Existing studies have shown that irisin is associated with promoting the conversion of white adipose tissue to brown adipose tissue, energy expenditure, glucose and lipid metabolism, and insulin resistance. A study by Noha M. Dawoud MD et al. (Circulating and / or cutaneous irisin resistance: A novel link among androgenetic alopecia, comorbid metabolic syndrome and cardiovascular risks, Journal of Cosmetic Dermatology, 2584) found elevated irisin levels in the serum of patients with androgenetic alopecia and was associated with various diseases. However, no treatment for hair loss was disclosed. Summary of the Invention

[0009] With regard to the bottleneck problem in the treatment of hair loss mentioned in the above background technology, the present invention aims to provide a class of polypeptides and their related applications for combating problems such as hair loss.

[0010] In one aspect, the present invention provides a use of a polypeptide, a polypeptide-derived peptide, or a polypeptide and a polypeptide-derived peptide derivative in the preparation of a medicament for treating seborrheic follicle decline and / or hair loss, seborrheic acne and / or dermatitis, or drug-induced acne and / or dermatitis.

[0011] The polypeptide is irisin;

[0012] The polypeptide-derived peptide is selected from irisin-derived peptides, which are conserved sequences between adjacent invertase cleavage sites in irisin;

[0013] The polypeptide and the derivative of the polypeptide-derived peptide are products obtained by conventional modification of the amino acid side chain groups, amino terminal end, and carboxyl terminal end of the irisin-derived peptide or irisin, or are products obtained by attaching a tag for polypeptide or protein detection or purification to the polypeptide, or are products obtained by isotope-labeled modification.

[0014] Furthermore, the irisin-derived peptide structural formula is a sequence represented by AAs1-X-AAs2 or a sequence represented by Za, wherein:

[0015] AAs1 and AAs2 are each independently a single or multiple amino acid polypeptide segment or absent;

[0016] X is His-Val-Gln-X1-Ile-Ser-X2-X3-Gly-X4-Ser-Pro-X5-Ser-X6-Pro-Val-Leu

[0017] X1 is Ala or Ser,

[0018] X2 is Ile or Met,

[0019] X3 is Gln or Ser or Gly,

[0020] X4 is Gln or Thr or Ser,

[0021] X5 is Ala or Pro or Leu,

[0022] X6 is Glu or Gln.

[0023] Further, wherein X is selected from one of:

[0024] HVQAISIQGQSPASEPVL SEQ ID No.1

[0025] HVQAISIQGQSPASEPLL SEQ ID No.2

[0026] HVQAISIQGQSPASEPVT SEQ ID No.3

[0027] HVQSISIQGQSPASEPVL SEQ ID No.4

[0028] HVQSISIQGQSPASEPVH SEQ ID No.5

[0029] HVQSISMSGTSPLSEPLR SEQ ID No.6

[0030] HVQSISMSGMSPLSEPLR SEQ ID No.7

[0031] Furthermore, M1 does not exist, and M2 is F, or both M1 and M2 do not exist.

[0032] Among them, SEQ ID NO.1 is a human irisin-derived peptide, SEQ ID NO.2 is a mouse irisin-derived peptide, SEQ ID NO.3 is a marsupial irisin-derived peptide, SEQ ID NO.4 is an avian irisin-derived peptide, SEQ ID NO.5 is a reptilian irisin-derived peptide, SEQ ID NO.6 is a bony fish-small yellow croaker irisin-derived peptide, and SEQ ID NO.7 is a bony fish-eel irisin-derived peptide.

[0033] Furthermore, in view of the functional similarities of irisin-derived peptides from different species, the irisin-derived peptides include irisin-derived peptides from mammals, reptiles, amphibians, bony fish, poultry, and birds.

[0034] Furthermore, the polypeptide-derived peptide is a conserved fragment of irisin derived from different species of organisms, and the polypeptide-derived peptide is located at the same position as the conserved fragment of the sequence described in any one of SEQ ID NOs. 1-7, and the different species of organisms are selected from mammals, reptiles, amphibians, bony fish, poultry and avian species, and the polypeptide-derived peptide is a sequence of X1-X2-...X n , and the upstream sequence and downstream sequence immediately adjacent to the polypeptide-derived peptide in irisin are both invertase cleavage sites, X represents an amino acid, and n is any integer from 8 to 25;

[0035] Furthermore, the conventional modifications include fluorescent group modification, phosphorylation modification, disulfide bond cyclization modification, biotin labeling modification, photosensitizer, azide modification, PEG modification, methylation modification, fluorescence quenching group modification, protein coupling modification, small molecule compound modification, amino modification, amidation modification, hydroxylation modification, carboxylation modification, carbonylation modification, alkylation modification, acetylation modification, esterification modification, and glycosylation modification;

[0036] The tag is His6, GST, EGFP, MBP, Nus, HA, IgG, FLAG, c-Myc or ProfinityeXact;

[0037] The modified product is selected from a product modified by a fluorescent group, a product modified by phosphorylation, a product modified by disulfide bond-based cyclization, a product modified by biotin labeling, a product modified by a photosensitizer, a product modified by azide, a product modified by PEG, a product modified by methylation, a product modified by a fluorescence quenching group, a product modified by protein coupling, or a product modified by a small molecule compound; or a product modified by amino modification, amidation modification, hydroxylation modification, carboxylation modification, carbonylation modification, alkylation modification, acetylation modification, esterification modification, or glycosylation modification.

[0038] Furthermore, the modification of the amino acid terminus is selected from acetylation modification of the polypeptide N-terminus, modification of the carboxyl terminus and amination modification of the C-terminus.

[0039] Furthermore, the side chain modification is selected from the modification of the R group of the amino acid side chain in the polypeptide.

[0040] Furthermore, the fluorescent dye used in the modification of the fluorescent group is selected from AMCA, FITC, Rhodamine, Cy3, Cy5, Cy5.5, Cy7, AIE, and ICG, and the modification can be used for fluorescence detection.

[0041] Furthermore, the phosphorylation modification is selected from one or more combinations of p-Ser, p-Thr, and p-Tyr.

[0042] Furthermore, the glycosylation modification is selected from a combination of one or more of Ser, Asn, Thr, and Tyr.

[0043] Furthermore, the nitration modification is selected from one or more combinations of Tyr.

[0044] Furthermore, the biotin label is selected from D-biotin, biotin hydrazide, photosensitive biotin and biotin-dUTP.

[0045] Furthermore, the photosensitizer modification can be used to prepare photosensitizing preparations.

[0046] Furthermore, the azide modification can be used for a secondary ligation reaction.

[0047] Furthermore, the PEG modification can be used to prepare drug carriers.

[0048] Furthermore, the isotope used in the isotope labeling is selected from one or more combinations of 13C, 14C, 14N, 15N, 2H, 3H, 18O, 32P, 32S, 34S, 35S, 36S, 35Cl, 37Cl, 125I, and 131I.

[0049] The peptides and derivatives based on them can be synthesized independently under standard chemical laboratory conditions. These can be recombinant proteins containing the peptides expressed from genes, or they can be synthesized industrially by commercial reagent companies using solid-phase methods, where different amino acids are condensed on a resin to form amino acid chains, or extracted from tissues and organs of different species. Peptide derivatives are obtained by attaching amino acids and then labeling them with modifying groups.

[0050] Furthermore, the hair follicle regression and / or hair loss caused by the drug is hair follicle regression and / or hair loss caused by anti-tumor drugs, immunosuppressants, arsenic agents, thallium acetate, coumarin, daunorubicin, clofibrate, allopurinol, phenindione, carbimazole, thiouracil, phenytoin sodium, ethambutol, indomethacin, levodopa, methyldopa, nitrofurantoin, nitrofurantoin, iodine, ethionamide, and gentamicin.

[0051] Furthermore, the anti-tumor drug is selected from nitrogen mustard anti-tumor drugs, such as nitrogen mustard hydrochloride, nitrogen mustard oxychloride, cyclophosphamide, chlorambucil, and carmustine; sulfonate anti-tumor drugs, such as busulfan; antimetabolites, such as methotrexate and fluorouracil; anti-tumor antibiotics, such as bleomycin and actinomycin; plant alkaloids, such as vinblastine and colchicine;

[0052] Furthermore, the seborrheic follicle decline and / or hair loss is seborrheic follicle decline and / or hair loss in females or males;

[0053] Drug-induced hair follicle decline and / or hair loss is drug-induced hair follicle decline and / or hair loss in females and males;

[0054] Seborrheic acne and / or dermatitis refers to seborrheic acne and / or dermatitis in females and males.

[0055] Furthermore, acne and / or dermatitis include problems in which the diameter of hair follicles becomes smaller, the diameter of sebaceous glands becomes larger, and skin cells or tissues have an inflammatory response.

[0056] Another aspect of the present invention provides the use of the above-mentioned polypeptide and its derivatives in the preparation of medical and cosmetic products, cosmetics, health products, foods, and additives for combating, preventing and / or treating hair follicle decline, hair loss, acne, or dermatitis.

[0057] Furthermore, the hair loss includes androgenic hair follicle decline and / or hair loss in males and females, and drug-induced hair follicle decline and / or hair loss;

[0058] Furthermore, the prevention and / or treatment of hair loss is promoting hair growth;

[0059] Furthermore, the acne includes male and female androgenic acne;

[0060] Furthermore, the dermatitis includes male and female androgenic dermatitis.

[0061] In another aspect, the present invention provides a drug for combating, preventing and / or treating hair follicle decline and hair loss, wherein the drug comprises the above-mentioned polypeptide, polypeptide-derived peptide or derivatives of the polypeptide and polypeptide-derived peptide.

[0062] Furthermore, the drug contains one or more pharmaceutically acceptable carriers;

[0063] The carrier is preferably a diluent, excipient, filler, binder, wetting agent, disintegrant, absorption promoter, adsorption carrier, surfactant or lubricant;

[0064] The medicine is an external preparation, which is selected from a smear, an ointment, a spray, a patch, a microneedle or an injection. The medicines in various dosage forms can be prepared according to conventional methods in the pharmaceutical field.

[0065] Another aspect of the present invention provides a method for preventing and / or treating hair follicle decline and hair loss, which comprises applying the above-mentioned polypeptide, polypeptide-derived peptide or polypeptide and polypeptide-derived peptide derivatives to the skin surface containing hair follicles.

[0066] Furthermore, the method is for non-diagnostic and non-therapeutic purposes.

[0067] Beneficial effects of the present invention:

[0068] (1) The polypeptide provided by the present invention and the derived polypeptide with the polypeptide as the skeleton can combat problems such as hair follicle decline and / or hair loss caused by androgens and / or chemotherapy drugs.

[0069] (2) The candidate polypeptides provided by the present invention and the derived polypeptides based on them are easy to synthesize and modify, are low-cost, and can be directly administered to the skin, thus having broad application prospects.

[0070] (3) The candidate polypeptides provided by the present invention and the derived polypeptides using the candidate polypeptides as the backbone provide a wide range of strategies for the preparation of diagnostic and therapeutic compounds or drugs for their practical application.

[0071] (4) The candidate polypeptides provided by the present invention and the derived polypeptides using the candidate polypeptides as the backbone are suitable for use in the treatment of hair loss and related symptoms. BRIEF DESCRIPTION OF THE DRAWINGS

[0072] Figure 1 This is a diagram showing the effect of the mouse-derived peptide (CY5-I001) linked to the dye CY5 in targeting the skin of female mice in vivo. Figure 1A shows the result of in vivo imaging of CY5-I001 enrichment in skin tissue after injection into the mouse tail vein. Figure 1 B is the staining result of frozen sections of mouse skin tissue after in vivo imaging, where the red signal represents the I001 polypeptide and the blue represents the cell nucleus stained with DAPI.

[0073] Figure 2 The therapeutic effect of I001 polypeptide in the female mouse androgenic (seborrheic) alopecia model. Figure 2 A is the hair growth on the back of the neck of female mice in the control group, androgen group, and androgen + polypeptide treatment group; Figure 2 B is the skin color scoring result of female mice.

[0074] Figure 3 These are the quantitative results of the skin morphology and hair follicle and sebaceous gland diameters on the back of the neck of female mice with androgenic (seborrheic) alopecia model. Figure 3 A is the HE staining results of the skin sections on the back of the neck of female mice in the control group, DHT treatment group and DHT+I001 polypeptide treatment group; Figure 3 B and Figure 3 C is based on Figure 3 A. Statistical results of the diameters of hair follicles and sebaceous glands in the skin after morphological observation.

[0075] Figure 4 The RNA-seq sequencing results of skin tissue of female mice with androgenic (seborrheic) alopecia model after treatment with I001 polypeptide were compared with those of female mice with model alopecia. Figure 4 A is a heat map; Figure 4 B is the GO function enrichment result of TOP30 differentially expressed genes; Figure 4 C is the KEGG pathway enrichment result of the TOP20 differentially expressed genes.

[0076] Figure 5 The therapeutic effect of I001 polypeptide in the androgenic (seborrheic) alopecia model in male rats. Figure 5 A is the hair growth on the back of the neck of male mice in the control group, androgen group and androgen + polypeptide treatment group; Figure 5 B is the skin color scoring result of male mice.

[0077] Figure 6 These are the quantitative results of skin morphology and diameter of hair follicles and sebaceous glands on the back of the neck of male mice with androgenic (seborrheic) alopecia model. Figure 6 A is the HE staining results of the skin sections on the back of the neck of male mice in the control group, DHT treatment group and DHT+I001 polypeptide treatment group; Figure 6 B and Figure 6 C is based on Figure 6 A. Statistical results of the diameters of hair follicles and sebaceous glands in the skin after morphological observation.

[0078] Figure 7The RNA-seq sequencing results of skin tissue of male mice with androgenic (seborrheic) alopecia model after treatment with I001 polypeptide were compared with those of male mice with model alopecia. Figure 7 A is a heat map; Figure 7 B is the GO function enrichment result of TOP30 differentially expressed genes; Figure 7 C is the KEGG pathway enrichment result of the TOP20 differentially expressed genes.

[0079] Figure 8 The therapeutic effect of I001 polypeptide in the drug-induced (CTX) hair loss model in female mice. Figure 8 A is the hair growth on the back of the neck of female mice in the control group, CTX (LD, low concentration) group, CTX (HD, high concentration) group, CTX (LD, low concentration) + I001 polypeptide treatment group, and CTX (HD, high concentration) + I001 polypeptide treatment group; Figure 8 B is the skin color scoring results of female mice in each group.

[0080] Figure 9 These are the RNA-seq sequencing results of skin tissues of female mice in a drug-induced (CTX, LD, low concentration) hair loss model after treatment with I001 polypeptide compared with female mice in the model model. Figure 9 A is a heat map; Figure 9 B is the GO function enrichment result of TOP30 differentially expressed genes; Figure 9 C is the KEGG pathway enrichment result of the TOP20 differentially expressed genes.

[0081] Figure 10 The therapeutic effect of I001 polypeptide in the male rat drug-induced (CTX) hair loss model. Figure 10 A is the hair growth on the back of the neck of male mice in the CTX group and the CTX+I001 polypeptide treatment group; Figure 10 B is the skin color scoring results of the two groups of male mice.

[0082] Figure 11 The RNA-seq sequencing results of skin tissue of male mice with drug-induced (CTX) alopecia model after treatment with I001 peptide were compared with those of male mice with model alopecia. Figure 11 A is a heat map; Figure 11 B is the GO function enrichment result of TOP30 differentially expressed genes; Figure 11 C is the KEGG pathway enrichment result of the TOP20 differentially expressed genes.

[0083] Figure 12 Identification results of primary human dermal papilla cells. Figure 12 A is a schematic diagram of the dermal papilla cells and their location and bright field morphological observation of cell culture; Figure 12 B is the identification result of primary human dermal papilla cells, and the green color is vimentin.

[0084] Figure 13 This study investigates the alleviating effects of homologous sequence peptides from different species on the changes in cell-related molecular markers after human primary dermal papilla cells were treated with DHT to induce an androgen / seborrheic model in vitro. Figure 13 A represents the effects of different peptides on androgen receptor AR and transforming growth factor TGFβ; Figure 13 B shows the effects of different peptides on Noggin, Wnt signaling pathway regulatory protein Wnt5a, Wnt signaling pathway regulatory protein β-catenin, fibroblast growth factor FGF7, and hair follicle stem cell marker CD133. DETAILED DESCRIPTION

[0085] In order to better understand the content of the present invention, the content of the present invention is further described below in conjunction with specific implementation cases, but the protection content of the present invention is not limited to the following embodiments.

[0086] In the present invention, the polypeptides and polypeptide-derived peptides are all derived from different species and are located in a conserved sequence polypeptide in irisin, wherein the sources of SEQ ID NO.1-7 are as follows: SEQ ID NO.1 is a human irisin-derived peptide, SEQ ID NO.2 is a mouse irisin-derived peptide, SEQ ID NO.3 is a marsupial irisin-derived peptide, SEQ ID NO.4 is an avian irisin-derived peptide, SEQ ID NO.5 is a reptile irisin-derived peptide, SEQ ID NO.6 is a bony fish-small yellow croaker irisin-derived peptide, and SEQ ID NO.7 is a bony fish-eel irisin-derived peptide.

[0087] The polypeptide-derived peptide is a conserved fragment of irisin derived from different species of organisms other than the above SEQ ID NOs. 1-7. The polypeptide-derived peptide is located at the same position as the conserved fragment of any of the sequences described in SEQ ID NOs. 1-7. The different species of organisms are selected from mammals, reptiles, amphibians, bony fish, poultry and avian species. The polypeptide-derived peptide is a sequence of X1-X2-...X n , and the immediately upstream sequence and downstream sequence of the polypeptide-derived peptide in irisin are both invertase cleavage sites, X represents an amino acid, and n is any integer from 8 to 25.

[0088] There are two conserved sequences in irisin, and the polypeptides and polypeptide-derived peptides in the present invention are all conserved sequences close to the N segment.

[0089] Example 1: Preparation of polypeptide sequences

[0090] The peptide sequence was synthesized by artificial synthesis.

[0091] The polypeptide sequence is as follows:

[0092] HVQAISIQGQSPASEPVL SEQ ID No.1 (human)

[0093] HVQAISIQGQSPASEPLL SEQ ID No. 2 (mouse I001)

[0094] HVQAISIQGQSPASEPVT SEQ ID No. 3 (marsupial-polypeptide)

[0095] HVQSISIQGQSPASEPVL SEQ ID No. 4 (bird-polypeptide)

[0096] HVQSISIQGQSPASEPVH SEQ ID No. 5 (Reptiles-polypeptide)

[0097] HVQSISMSGTSPLSEPLR SEQ ID No.6 (bony fish - small yellow croaker)

[0098] HVQSISMSGMSPLSEPLR SEQ ID No.7 (bony fish - yellow eel)

[0099] The peptides were synthesized using conventional solid-phase or liquid-phase synthesis methods. The solid-phase peptide synthesis method involves reacting the amino acid from the C-terminus to the N-terminus, followed by resin activation, amino acid linking, elution and protection, and detection, completing the amino acid linking step by step. The peptides were then precipitated and centrifuged with excess ether. The crude peptides were purified by HPLC, analyzed by mass spectrometry, and then freeze-dried in liquid nitrogen for later use.

[0100] SEQ ID NO.1 is a human irisin-derived peptide, SEQ ID NO.2 is a mouse irisin-derived peptide, SEQ ID NO.3 is a marsupial irisin-derived peptide, SEQ ID NO.4 is an avian irisin-derived peptide, SEQ ID NO.5 is a reptilian irisin-derived peptide, SEQ ID NO.6 is a teleost-small yellow croaker irisin-derived peptide, and SEQ ID NO.7 is a teleost-eel irisin-derived peptide.

[0101] Many proteins and hormones share a high degree of homology between humans and animals. For example, bovine and porcine insulin can be used to treat diabetes; pregnant mare serum gonadotropin (PMSG) has also been used for superovulation in humans and animals; the chemical structure of oxytocin in humans and most mammals is the same; and since gonadotropin-releasing hormone (GnRH) was isolated from pig and sheep brains in 1971, the GnRH family has at least 28 types, 15 of which come from vertebrates and 13 from invertebrates. With the exception of octopus GnRH, all GnRH peptides are composed of 10 amino acids, and their molecular length and partial amino acid sequence are highly conserved.

[0102] Given the homology and functional similarity between polypeptides and hormones in different species, sequences from the same species or with higher homology to the polypeptide may play the same role.

[0103] Example 2: Preparation of polypeptide probes

[0104] This example provides a polypeptide probe, which uses the polypeptide prepared in Example 1 and combines it with Cy5 through an organic chemical reaction. The specific method is to entrust a company to synthesize it.

[0105] Example 3: Detection of the ability of peptide Cy5 probe to target female mouse skin

[0106] Adult female mice were injected with the Cy5-labeled polypeptide prepared in Example 2 and PBS via the tail vein, with each mouse receiving 100 μL of the injection. 24 hours later, the cells were detected using a small animal imaging device.

[0107] The experimental results are shown in Figure 1 ,pass Figure 1 A shows that the skin tissue has obvious fluorescence signals compared with the control group, indicating that I-001 (SEQ ID NO. 2) has a receptor that binds to it on the skin; further Figure 1 The staining results of the frozen section of skin tissue in B show that I-001 is enriched in multiple cell types of the skin, including epidermal cells, fibroblasts, hair follicle cells, muscle cells, etc., suggesting that I001 exerts biological functions on these cells.

[0108] Example 4: Mouse androgenic (seborrheic) alopecia model

[0109] Six-week-old male and female C57BL / 6 mice were divided into a control group, a DHT intraperitoneal injection group, and a DHT intraperitoneal injection + polypeptide application group. DHT (2 mg / d) was administered every other day.

[0110] Six days after the first DHT treatment, hair removal was performed (removing 2 x 2 cm of telogen hair on the back and using a depilatory cream) to induce synchronous hair regrowth. The peptide application concentration was 0.162232317M, and the dosage was 0.1 ml.

[0111] The experimental results are shown in Figure 2 and Figure 5 , Figure 2 This is a female mouse androgenic (seborrheic) alopecia model. Figure 5 This is a model of androgenic (seborrheic) alopecia in male mice. Comparison of the results shows that the I001 polypeptide can treat androgen-induced alopecia in both male and female mice, with significant effects seen starting on day 10 in female mice and on day 12 in male mice.

[0112] Furthermore, HE staining results of the skin sections of the back of the neck of each group of mice ( Figure 3 A, Figure 6 A) It can be seen that after DHT treatment, hair follicles atrophy and sebaceous glands hypertrophy, which were significantly alleviated after peptide treatment; quantitative results showed that after DHT treatment, the diameter of hair follicles was significantly reduced and the diameter of sebaceous glands was significantly increased, and the diameter of hair follicles and sebaceous glands was significantly restored after peptide treatment (Figure Figure 3 B / C, Figure 6 B / C).

[0113] Furthermore, the RNA-seq sequencing results of the neck and back skin of mice in the peptide treatment group and the DHT model group were analyzed ( Figure 4 Female mice, Figure 7 Male mice) showed that the peptide improved DHT-induced hair loss in female mice mainly by regulating immune and inflammatory responses, responses to bacteria, keratinized fibers, skin epidermal structure composition, immune and chemotaxis-related signaling pathways ( Figure 4 B, C), in male mice, it improves DHT-induced hair loss mainly by regulating steroid hormones, cell response to fatty acids, and the synthesis and secretion of growth hormone ( Figure 7 B, C).

[0114] Acne is a chronic inflammatory skin disease of the hair follicles and sebaceous glands that is common in adolescents and adults. Its currently recognized mechanisms are mainly four aspects: excessive keratinization of the epithelium of hair follicles and sebaceous gland ducts; excessive secretion of sebaceous glands; colonization of microorganisms (Propionibacterium acnes and Malassezia); and inflammation and immune response.

[0115] Sequencing results showed that I001 peptide treatment could improve the damage caused by DHT model to mouse skin in multiple aspects such as keratinization, fatty acids, immunity and bacterial response. It is inferred that I001 peptide has a certain therapeutic effect on hyperandrogen-induced dermatitis and acne.

[0116] Example 5: Mouse drug-induced alopecia model

[0117] Seven-week-old C57BL / 6 mice in the telogen phase were depilated on day 0 to induce hair synchronization. Nine days after depilation, all hair follicles were in anagen phase VI. At this point, mice were intraperitoneally injected with a low-dose 50 mg / kg or high-dose 150 mg / kg cyclophosphamide for induction. The peptide was topically administered starting on day 6, at the same concentration of 0.162232317 M, in a 0.1 ml volume.

[0118] On the 13th day, it can be seen that in the low-concentration group of female mice, the peptide effectively alleviated the CTX-induced hair loss phenomenon, while the high-concentration group had no significant difference ( Figure 8 Therefore, male mice were only induced with low concentrations of CTX. It can be seen that on day 13, the peptide was also able to alleviate the CTX-induced hair loss phenomenon in male mice ( Figure 10 ).

[0119] Furthermore, the RNA-seq sequencing results of the mouse neck and back skin were analyzed ( Figure 9 Female mice, Figure 11 Male mice) showed that the peptide improved CTX-induced hair loss in female mice mainly by regulating keratinization reaction, keratinized fibers, skin epidermal structure composition and oxidative phosphorylation ( Figure 9 B, C), in male mice, CTX-induced hair loss was improved mainly by regulating lipid metabolism, keratinization reaction, epithelial cell differentiation, keratinized fibers, skin epidermal structure composition, biosynthesis of unsaturated fatty acids and IL-17 inflammatory signaling pathway ( Figure 11 B, C).

[0120] Example 6: Purchase and identification of primary human dermal papilla cells

[0121] Human primary dermal papilla cells were purchased from Shanghai Xuanke Biotechnology Co., Ltd. The cells were derived from normal scalp tissue removed by surgery, and the immunofluorescence staining of vimentin was positive ( Figure 12 B) The cells are certified to be greater than 90% pure and free of HIV-1, HBV, HCV, mycoplasma, bacteria, yeast, and fungi. The culture medium is fibroblast culture medium.

[0122] Example 7: DHT-induced androgen / seborrheic alopecia cell model

[0123] After serum starvation for 24 hours, human primary dermal papilla cells were treated with high androgen levels using 10 nM DHT. The expression of cell markers (AR, TGF-β, Noggin, Wnt5a, β-Catenin, Fgf7, CD133, etc.) was measured 24 hours later. Homologous peptides from different species (the peptides described in Example 1) were also added to the culture medium, and the expression of cell markers was measured 24 hours later.

[0124] like Figure 13 The results showed that the expression of androgen receptor AR and the inflammatory factor TGF-β in hair follicle cells increased significantly after DHT treatment ( Figure 13 A), while the expression of Wnt signaling pathway regulatory proteins Wnt5a and β-Catenin decreased, as did the hair follicle stem cell marker CD133, Noggin, and Fgf7. Peptides with homologous sequences from different species (P1-P7 corresponding to SEQ ID NOs. 1-7, respectively) were able to significantly alleviate these changes. Activation of the Wnt / β-catenin signaling pathway plays a central role in hair follicle regeneration; Fgf7 is a dermal papilla signal that instructs hair germ cells to proliferate and initiate a new hair cycle; and Noggin is a gene that shortens the resting phase and initiates the growth phase of new hair.

[0125] It is shown that the polypeptide of the present invention can be used to alleviate androgenic alopecia, especially androgenic / seborrheic follicle decline or alopecia caused by significantly increased expression of androgen receptor AR, apoptosis of hair follicle cells, abnormally increased expression of inflammatory factors, and hair follicle degeneration, as well as androgenic / seborrheic acne or dermatitis.

[0126] In summary, the polypeptide and its related substances provided in this application can be used to prepare therapeutic drugs for combating diseases such as hair follicle decline and / or hair loss caused by androgens, and / or seborrheic dermatitis, and / or drugs.

[0127] The above description is only a specific embodiment of the present invention, not all embodiments. Any equivalent transformation of the technical solution of the present invention made by ordinary technicians in this field after reading the specification of the present invention is covered by the claims of the present invention.

Claims

1. Use of a polypeptide, a polypeptide-derived peptide, or a polypeptide and a polypeptide-derived peptide derivative in the preparation of a medicament for treating seborrheic follicular atrophy and / or alopecia, drug-induced follicular atrophy and / or alopecia, seborrheic acne, and / or dermatitis; The polypeptide is irisin; The polypeptide-derived peptide is selected from irisin-derived peptides, which are conserved sequences between adjacent invertase cleavage sites in irisin; The polypeptide and the derivative of the polypeptide-derived peptide are products obtained by conventional modification of the amino acid side chain groups, amino terminal end, and carboxyl terminal end of the irisin-derived peptide or irisin, or are products obtained by attaching a tag for polypeptide or protein detection or purification to the polypeptide, or are products obtained by isotope-labeled modification.

2. The use according to claim 1, characterized in that The irisin-derived peptide structural formula is a sequence represented by AAs1-X-AAs2 or a sequence represented by Za, wherein: AAs1 and AAs2 are each independently a single or multiple amino acid polypeptide segment or absent; X is His-Val-Gln-X1-Ile-Ser-X2-X3-Gly-X4-Ser-Pro-X5-Ser-X6-Pro-Val-Leu X1 is Ala or Ser, X2 is Ile or Met, X3 is Gln or Ser or Gly, X4 is Gln or Thr or Ser, X5 is Ala or Pro or Leu, X6 is Glu or Gln; Preferably, X is selected from one of: HVQAISIQGQSPASEPVL SEQ ID No.1 HVQAISIQGQSPASEPLL SEQ ID No.2 HVQAISIQGQSPASEPVT SEQ ID No.3 HVQSISIQGQSPASEPVL SEQ ID No.4 HVQSISIQGQSPASEPVH SEQ ID No.5 HVQSISMSGTSPLSEPLR SEQ ID No.6 HVQSISMSGMSPLSEPLR SEQ ID No.7 More preferably, M1 is absent, and M2 is F, or both M1 and M2 are absent.

3. The use according to claim 1, characterized in that The irisin-derived peptide is a conserved sequence between adjacent invertase cleavage sites in irisin from mammals, reptiles, amphibians, bony fish, birds, and avians; Preferably, the irisin-derived peptide is located at the same conserved fragment position as the sequence of any one of SEQ ID NOs. 1-7; Preferably, the irisin-derived peptide comprises 8-25 amino acids.

4. The use according to claim 1, characterized in that The hair follicle regression and / or hair loss caused by the drug is hair follicle regression and / or hair loss caused by anti-tumor drugs, immunosuppressants, arsenic agents, thallium acetate, coumarin, daunorubicin, clofibrate, allopurinol, phenindione, carbimazole, thiouracil, phenytoin sodium, ethambutol, indomethacin, levodopa, methyldopa, nitrofurantoin, nitrofurantoin, iodine, ethionamide, and gentamicin; Preferably, the antitumor drug is selected from nitrogen mustard antitumor drugs, sulfonate antitumor drugs, antimetabolites, antitumor antibiotics, and plant alkaloids.

5. The use according to claim 1, characterized in that The seborrheic follicle decline and / or hair loss is seborrheic follicle decline and / or hair loss in females or males; Drug-induced hair follicle decline and / or hair loss is drug-induced hair follicle decline and / or hair loss in females and males; Seborrheic acne and / or dermatitis refers to seborrheic acne and / or dermatitis in females and males.

6. The use according to claim 1, characterized in that The conventional modifications include fluorescent group modification, phosphorylation modification, disulfide bond cyclization modification, biotin labeling modification, photosensitizer, azide modification, PEG modification, methylation modification, fluorescence quenching group modification, protein coupling modification, small molecule compound modification, amino modification, amidation modification, hydroxylation modification, carboxylation modification, carbonylation modification, alkylation modification, acetylation modification, esterification modification, and glycosylation modification; The tag is His6, GST, EGFP, MBP, Nus, HA, IgG, FLAG, c-Myc or ProfinityeXact; The modified product is selected from a product modified by a fluorescent group, a product modified by phosphorylation, a product modified by disulfide bond cyclization, a product modified by biotin labeling, a product modified by a photosensitizer, a product modified by azide, a product modified by PEG, a product modified by methylation, a product modified by a fluorescence quenching group, a product modified by protein coupling, or a product modified by a small molecule compound; or a product modified by aminoation, amidation, hydroxylation, carboxylation, carbonylation, alkylation, acetylation, esterification, or glycosylation. Preferably, the modification of the amino acid terminus is selected from the group consisting of acetylation modification of the N-terminus of the polypeptide, modification of the carboxyl terminus and amination modification of the C-terminus; Preferably, the side chain modification is selected from the modification of the R group of the amino acid side chain in the polypeptide; Preferably, the fluorescent dye used in the modification of the fluorescent group is selected from AMCA, FITC, Rhodamine, Cy3, Cy5, Cy5.5, Cy7, AIE, and ICG, and the modification can be used for fluorescence detection; Preferably, the phosphorylation modification is selected from a combination of one or more of p-Ser, p-Thr, and p-Tyr; Preferably, the glycosylation modification is selected from a combination of one or more of Ser, Asn, Thr, and Tyr; Preferably, the nitration modification is selected from one or more combinations of Tyr; Preferably, the biotin label is selected from D-biotin, biotin hydrazide, photosensitive biotin and biotin-dUTP; Preferably, the photosensitizer is modified and can be used to prepare a photosensitizing preparation; Preferably, the azide modification can be used for a secondary ligation reaction; Preferably, the PEG modification can be used to prepare a drug carrier; Preferably, the isotope used in the isotope labeling is selected from one or more combinations of 13C, 14C, 14N, 15N, 2H, 3H, 18O, 32P, 32S, 34S, 35S, 36S, 35Cl, 37Cl, 125I, and 131I.

7. Use of the polypeptide, polypeptide-derived peptide, or derivatives of the polypeptide and polypeptide-derived peptide according to any one of claims 1 to 6 in the preparation of medical and cosmetic products, cosmetics, health products, foods, and additives for combating, preventing, and / or treating hair follicle decline, hair loss, acne, or dermatitis; The hair follicle decline and hair loss include male and female androgenic hair follicle decline and / or hair loss, and drug-induced hair follicle decline and / or hair loss; Preferably, the prevention and / or treatment of hair loss is promoting hair growth; Preferably, the acne includes male and female androgenic acne; Preferably, the dermatitis includes male and female androgenic dermatitis.

8. The use according to claim 7, characterized in that The hair follicle regression and / or hair loss caused by the drug is hair follicle regression and / or hair loss caused by anti-tumor drugs, immunosuppressants, arsenic agents, thallium acetate, coumarin, daunorubicin, clofibrate, allopurinol, phenindione, carbimazole, thiouracil, phenytoin sodium, ethambutol, indomethacin, levodopa, methyldopa, nitrofurantoin, nitrofurantoin, iodine, ethionamide, and gentamicin; Preferably, the antitumor drug is selected from nitrogen mustard antitumor drugs, sulfonate antitumor drugs, antimetabolites, antitumor antibiotics, and plant alkaloids.

9. A drug for combating, preventing and / or treating hair follicle decline and hair loss, comprising the polypeptide, polypeptide-derived peptide, or derivatives of the polypeptide and polypeptide-derived peptide according to any one of claims 1 to 6; Preferably, the drug is an external preparation. More preferably, the external preparation is selected from a smear, an ointment, a spray, a patch or a microneedle.

10. A method for preventing and / or treating hair follicle atrophy, hair loss, acne or dermatitis, the method comprising applying the polypeptide, polypeptide-derived peptide or polypeptide and polypeptide-derived peptide derivative according to any one of claims 1 to 6 to the surface of skin containing hair follicles; the hair follicle atrophy or hair loss is caused by androgen or drugs; Preferably, the method is non-diagnostic and non-therapeutic.

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