Polypeptides for resisting alopecia and dermatitis, related substances and application thereof
Through the method of binding of peptides to GPR1 receptors, the side effects and insufficient application scope of existing hair loss treatment drugs have been solved, and effective treatment of hair loss, acne and dermatitis caused by stress and androgens has been achieved. It is suitable for a wide variety of people, especially women and middle-aged women, with low cost and wide application prospects.
Patent Information
- Application Number
- CN202310793536.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2023-06-30
- Publication Date
- 2024-12-31
AI Technical Summary
The existing hair loss treatment drugs have side effects and are not suitable for a wide variety of people or multiple types of hair loss, especially women and middle-aged women. The mechanism of androgenic hair loss is unclear in women and lacks effective interventions.
Provides a polypeptide and its derivatives that bind to GPR1 receptors and act on the hair follicles, skin and endocrine systems for the treatment and prevention of hair loss, acne and dermatitis caused by stress, androgens and seborrheicity, including through The modified and labeled peptide binds to the receptor, triggering downstream signaling pathways to inhibit inflammatory responses and promote hair growth.
The polypeptide and its derivatives are effective in combating hair follicle decline, hair loss, acne and dermatitis caused by stress and androgens. They are suitable for a wide variety of people, including women and middle-aged women, and are easy to synthesize and modify, and are inexpensive. They are suitable for a variety of dosage forms and Application scenario.
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Abstract
Description
Technical Field
[0001] The 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 application. Background Art
[0002] Hair loss (Hair Loss, Loss of hair) refers to the phenomenon of hair loss, which is divided into physiological hair loss and pathological hair loss. Among them, physiological hair loss refers to the normal loss of hair during the regression and resting phases to maintain a normal amount of hair. Pathological hair loss refers to abnormal or excessive hair loss.
[0003] The data from the 2017 "China Hair Loss Population Survey Report" shows that there are currently about 200 million people with hair loss in my country, and the number is increasing at a rate of at least 15% per year. The majority of people with hair loss are men, about 130 million.
[0004] There are many reasons for hair loss, including genetic factors, immune factors, excessive mental stress, staying up late, drugs (such as chemotherapy-induced hair loss), environmental factors, endocrine disorders, etc.
[0005] According to the cause, hair loss can be divided into stress alopecia, androgenic / seborrheic alopecia, pathological alopecia, physical / chemical alopecia, nutritional alopecia, obesity-related alopecia, hereditary alopecia, and scarring alopecia. 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 for scalp application, which has a "crazy shedding period", takes a long time to take effect, and has certain side effects; finasteride is an oral drug, but it has major hidden dangers to female fertility and is ineffective for postmenopausal women, so it is not recommended for women to take it. Clinically, it is necessary to develop safer, more effective, and intervention drugs suitable for a variety of populations and a variety of hair loss types.
[0006] Androgenic alopecia (AGA) is divided into male pattern (MPHL) and female pattern (FPHL), which is caused by genetic factors and the effects of androgens. The difference in sex hormones between men and women makes male and female hair loss very different, and there are differences in etiology, diagnosis and treatment. Men have higher levels of androgen in their bodies. After testosterone reaches the hair follicles, it will be converted into dihydrotestosterone DHT by 5α-reductase, causing hair follicle atrophy and hair gradually becoming thinner. In men with genetic factors, it causes hair loss, mainly manifested as a receding hairline, M-shaped hair loss, and finally baldness. For women, the level of androgen in the body is low, and estrogen can inhibit androgen. However, when the hormone balance in the body is broken or estrogen decreases after middle age, hair loss will occur due to insufficient estrogen secretion / excessive androgen, mainly showing progressive hair loss, with progressive hair follicle miniaturization as the clinical feature, and the hair in the center of the top of the head gradually becoming thinner and sparse.
[0007] 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 Diagnosis and Treatment of Female Androgenic Alopecia (2022 Edition)"]. Summary of the invention
[0008] With regard to the bottleneck problem of hair loss treatment mentioned in the above background technology, the purpose of the present invention is to provide a class of polypeptides and their related applications for combating hair loss and other problems.
[0009] On one hand, 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 drug for treating hair follicle decline and / or hair loss caused by stress, seborrheic hair follicle decline and / or hair loss, acne and / or dermatitis caused by stress, and seborrheic acne and / or dermatitis;
[0010] The polypeptide is selected from the polypeptide sequence shown in any one of the following SEQ ID NO.1-9:
[0011]
[0012] The polypeptide-derived peptide is a polypeptide comprising the above sequence, or a sequence having a homology higher than 50% with the above sequence.
[0013] The derivatives are products obtained by modifying the terminal or side chain of the above-mentioned polypeptide or polypeptide-derived peptide, or products obtained by connecting a tag for polypeptide or protein detection or purification, or products obtained by isotope labeling modification;
[0014] 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, a product modified by a small molecule compound, or a product modified by aminoation, amidation, hydroxylation, carboxylation, carbonylation, alkylation, acetylation, esterification, or glycosylation.
[0015] Furthermore, the terminal modification is selected from the group consisting of an acetylation modification at the N-terminus of the polypeptide and an amidation modification at the C-terminus.
[0016] Furthermore, the modification of the side chain is selected from the modification of the R group of the amino acid side chain in the polypeptide.
[0017] 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.
[0018] Furthermore, the phosphorylation modification is selected from a combination of one or more of p-Ser, p-Thr, and p-Tyr.
[0019] Furthermore, the glycosylation modification is selected from a combination of one or more of Ser, Asn, Thr, and Tyr.
[0020] Furthermore, the nitration modification is selected from one or more combinations of Tyr.
[0021] Furthermore, the biotin label is selected from D-biotin, biotin hydrazide, photosensitive biotin and biotin-dUTP.
[0022] Furthermore, the photosensitizer modification can be used to prepare photosensitizers.
[0023] Furthermore, the azide modification can be used for a secondary ligation reaction.
[0024] Furthermore, the PEG modification can be used to prepare drug carriers.
[0025] Furthermore, the isotopes used in the isotope labeling are selected from one or more combinations of 13C, 14C, 14N, 15N, 2H, 3H, 18O, 32P, 32S, 34S, 35S, 36S, 35Cl, 37Cl, 125I, and 131I.
[0026] The polypeptide and its derivatives with the backbone can be synthesized independently under general chemical laboratory conditions, through gene expression of recombinant proteins containing the polypeptide, or industrially synthesized by commercial reagent companies, using solid phase method to synthesize polypeptides, and directing the synthesis of amino acid chains through condensation reaction of different amino acids on the resin, or tissue extracts from different species. The derivatives of polypeptides are marked with modified groups after the amino acids are connected.
[0027] Further, the hair follicle decline and / or hair loss caused by stress is hair follicle decline and / or hair loss caused by stress in female or male subjects;
[0028] Seborrheic follicular atrophy and / or alopecia is seborrheic follicular atrophy and / or alopecia in male or female subjects;
[0029] Stress-induced acne and / or dermatitis is stress-induced acne and / or dermatitis in male or female subjects;
[0030] Seborrheic acne and / or dermatitis is seborrheic acne and / or dermatitis in male or female subjects;
[0031] Another aspect of the present invention provides the above polypeptide binding to its receptor and exerting its function.
[0032] Furthermore, the receptor mainly refers to GPR1.
[0033] Another aspect of the present invention provides the use of a substance capable of binding to and exerting its function on a GPR1 receptor in the preparation of a medicament for treating and / or preventing hair follicle decline and / or hair loss caused by stress, seborrheic hair follicle decline and / or hair loss, acne and / or dermatitis caused by stress, and seborrheic acne and / or dermatitis.
[0034] Furthermore, the substance capable of binding to the GPR1 receptor and exerting its function is a substance capable of binding to the GPR1 receptor and triggering its downstream signaling pathway;
[0035] Furthermore, the substance capable of binding to the GPR1 receptor and exerting its function is the above-mentioned polypeptide.
[0036] Furthermore, substances that can bind to the GPR1 receptor and exert their functions include, in addition to the above-mentioned polypeptides, inorganic ions such as cations / metal ions, anions, and hydrogen ions; organic molecules such as polypeptides and small molecule compounds; biological macromolecules such as proteins, such as enzymes, and screened receptor-binding peptides; antibodies, siRNAs, shRNAs, antisense RNAs, and DNA enzymes or combinations thereof, as well as expression vectors containing siRNAs, shRNAs, and antisense molecules.
[0037] Another aspect of the present invention provides the use of the above-mentioned polypeptides, polypeptide-derived peptides or polypeptides and polypeptide-derived peptide derivatives or substances that can bind to and function with the GPR1 receptor in the preparation of medical and beauty products, cosmetics, health products, foods, and additives for combating, preventing and / or treating hair follicle decline, hair loss, acne, and dermatitis.
[0038] Further, the hair loss includes stress-induced hair follicle decline and / or hair loss in males and females, androgenic hair follicle decline and / or hair loss;
[0039] Furthermore, the prevention and / or treatment of hair loss is promoting hair growth;
[0040] Further, the acne includes male and female stress acne and androgenic acne;
[0041] Furthermore, the dermatitis includes male and female stress dermatitis and androgenic dermatitis.
[0042] Another aspect of the present invention provides the use of the above polypeptide, polypeptide-derived peptide or polypeptide and polypeptide-derived peptide derivatives or substances capable of binding to and functioning with the GPR1 receptor in the preparation of detection reagents targeting the brain, skin, adrenal glands and gonadal systems.
[0043] Further, the brain is a female or male brain;
[0044] Further, the skin is female or male skin;
[0045] Further, the adrenal gland is a female or male adrenal gland;
[0046] Further, the male gonad system is the testis and epididymis;
[0047] Furthermore, the female gonadal system is ovaries.
[0048] In another aspect, the present invention provides a use of a substance capable of binding to a GPR1 receptor in the preparation of medicines, medical and cosmetic products, cosmetics, health products, foods, and additives for inhibiting the increase of corticosteroids, or inhibiting the inflammatory response and / or hair follicle decline and / or hair loss caused by the increase of corticosteroids;
[0049] Preferably, the substance capable of binding to the GPR1 receptor is the above-mentioned polypeptide, polypeptide-derived peptide or a derivative of the polypeptide and polypeptide-derived peptide.
[0050] In another aspect, the present invention provides a pharmaceutical composition for combating, preventing and / or treating symptoms related to hair follicle decline, hair loss, acne, dermatitis, etc., wherein the pharmaceutical composition comprises the above-mentioned polypeptide, polypeptide-derived peptide or polypeptide and polypeptide-derived peptide derivatives, and conjugates of various forms of agonists and antagonists of receptor binding and active ingredients.
[0051] The drug may contain one or more pharmaceutically acceptable carriers;
[0052] The carrier is preferably a diluent, excipient, filler, binder, wetting agent, disintegrant, absorption promoter, adsorption carrier, surfactant or lubricant;
[0053] The drug can be further prepared into various forms such as drops, tablets, granules, capsules, oral liquids, inhalation liquids, smears, sprays, microneedles or injections, and the drugs in various dosage forms can be prepared according to conventional methods in the pharmaceutical field;
[0054] The drug administration methods include oral administration, topical contact, administration as a suppository, intravenous, intraperitoneal, intramuscular, intralesional, intrathecal, intranasal or subcutaneous administration to the subject. The administration routes include parenteral administration such as intravenous, intramuscular, intraarterial, intradermal, subcutaneous, intraperitoneal, intraventricular and intracranial; transmucosal administration such as buccal, sublingual, palatal, gingival, nasal, vaginal, rectal or transdermal. Other delivery modes include, but are not limited to, the use of liposome preparations, intravenous infusion, transdermal patches, etc.
[0055] Beneficial effects of the present invention:
[0056] (1) The candidate polypeptides provided by the present invention and their derivatives with the backbones can combat problems such as hair follicle decline, hair loss, acne, and / or dermatitis caused by stress, and / or androgens, and / or seborrheic dermatitis.
[0057] (2) The candidate polypeptides and derivatives with the candidate polypeptides as the backbone provided by the present invention are easy to synthesize and modify, and are low in cost, and therefore have broad application prospects.
[0058] (3) The candidate polypeptides and derivatives based on the candidate polypeptides provided by the present invention provide a wide range of strategies for preparing diagnostic and therapeutic compounds or drugs for their practical applications.
[0059] (4) The candidate polypeptides provided by the present invention and their derivatives with the backbones are suitable for use in the treatment of hair loss and related symptoms, and are also suitable for use as research and observation tools in experimental animals for hair loss disease modeling. BRIEF DESCRIPTION OF THE DRAWINGS
[0060] Figure 1 This is a peptide-receptor binding experiment. Figure 1 A is the molecular docking simulation results of LRH-G3 with hGPR1 and mGPR1; Figure 1 B is the identification result of HEK293 cell lines overexpressing human GPR1 and mouse GPR1; Figure 1 C is the immunofluorescence staining result of LRH-G3 binding to HEK293-hGPR1 and HEK293-mGPR1 cells.
[0061] Figure 2 It is the effect of polypeptide after binding to receptor. Figure 2 A: Peptides LRH-G3 and LRH-G5 alleviate the inhibitory effect of chemerin on the cAMP signaling pathway; Figure 2 B is a peptide that inhibits the calcium (Ca) induced by chemerin 2+ )Inward flow effect.
[0062] Figure 3The expression and cellular localization results of the receptors of peptides and their derivatives in the human brain, testis and ovary are summarized through the single-cell sequencing results of the human brain, testis and ovary in The human Protein Atlas database.
[0063] Figure 4 The figure shows the effect of Cy5-labeled peptide (Cy5-G5 / G3) targeting the ovaries, adrenal glands, brain and skin of female mice in vivo. The figure shows the distribution and results of fluorescence signals of Cy5-labeled irrelevant peptides and LRH-G5 and LRH-G3 in the ovaries, adrenal glands, brain and skin of three groups of female mice.
[0064] Figure 5 The figure shows the effect of Cy5-labeled peptide (Cy5-G5 / G3) targeting the brain, gonads, adrenal glands and skin of male mice in vivo. The figure shows the distribution and results of fluorescence signals of Cy5-labeled irrelevant peptides and LRH-G5 and LRH-G3 in the brain, testis, epididymis, adrenal glands and skin of male mice.
[0065] Figure 6 The results of immunofluorescence staining of GPR1 in the hypothalamus of female mice and the co-localization of GPR1 with GnRH and CRF. Figure 6 A shows the expression of GPR1 in the mouse hypothalamus. The brown part is GPR1 and the blue part is the cell nucleus. Figure 6 B is a representative immunostaining image of CRF and GPR1 in the hypothalamus of female mice. Figure 6 C is a representative immunostaining image of GnRH and GPR1 in the hypothalamus of female mice. GPR1 can be seen to be co-localized with GnRH and CRF.
[0066] Figure 7 The results of immunofluorescence staining of GPR1 in the pituitary of female mice and the co-localization of GPR1 with pituitary hormones FSH / LH, GH, TSH and ACTH. The red part in the figure is the pituitary hormone, the green part is GPR1, and the blue part is the cell nucleus. It can be seen that the expression of GPR1 and pituitary hormones is co-localized.
[0067] Figure 8 The therapeutic effect of the peptide in the male rat stress-induced hair loss model. Figure 8 A is the hair growth on the back of the neck of mice in the control group, the steroid corticosteroid dexamethasone group, and the dexamethasone + peptide treatment group; Figure 8 B is the scoring standard for mouse skin color; Figure 8 C is Figure 8 The result of A is Figure 8 The results of the standard quantification of B.
[0068] Fig. 9The therapeutic effect of the peptide in the androgenic (seborrheic) alopecia model in female mice. Fig. 9 A is the hair growth on the back of the neck of mice in the control group, androgen (DHT) group and androgen + polypeptide treatment group; Fig. 9 B is the scoring standard for mouse skin color; Fig. 9 C is Fig. 9 The result of A is Fig. 9 The results of the standard quantification of B.
[0069] Fig.10 The therapeutic effect of the peptide in the male rat androgenic (seborrheic) alopecia model. Fig.10 A is the hair growth on the back of the neck of mice in the androgen (DHT) group and the androgen + polypeptide treatment group; Fig.10 B is the scoring standard for mouse skin color; Fig.10 C is Fig.10 The result of A is Fig.10 The results of the standard quantification of B.
[0070] Fig.11 The growth of hair on the back of GPR1 knockout (GPR1 KO) male mice after application of the androgen DHT was compared with that of wild-type male mice. Fig.11 A is the back hair growth of wild-type and GPR1 KO male mice when the androgen DHT was applied on different days; Fig.11 B is the scoring standard for mouse skin color; Fig.11 C is Fig.11 The result of A is Fig.11 The results of the standard quantification of B.
[0071] Fig.12 The following are the gene expressions of androgen receptor AR and related enzymes, 5α-reductase (the enzyme that converts testosterone into dihydrotestosterone DHT), testosterone production-related enzymes 3βHSD and 17βHSD in the back skin tissue of wild-type and GPR1 KO male mice after 14 days of DHT application; gene expressions of hair follicle cell apoptosis factor TGFβ, inflammatory factors TNFα and IL-6; gene expression of hair follicle degeneration marker MMP-9; gene expression of Wnt signaling pathway-related factor Wnt7a and its antagonist DKK1; gene expression of fat differentiation factor PPARγ and fatty acid binding protein FABP3. In the figure, each group of bar graphs represents WT-ctl, WT-DHT, GPR1 KO-ctl, and GPR1 KO-DHT, respectively.
[0072] Fig.13The G1-G6 sequence peptides alleviated the changes of cell-related molecular markers after the androgen / seborrheic model was induced by DHT treatment using human primary dermal hair papilla cells in vitro. Among them, AR is the androgen receptor, 5α-reductase promotes the conversion of testosterone into dihydrotestosterone DHT, 3βHSD and 17βHSD are hormone synthesis-related enzymes; Caspase-3, Bax, Bcl-2 are markers of hair follicle cell apoptosis; CD200 and CD133 are markers of hair follicle stem cells, MMP-9 is a marker of hair follicle degeneration; Wnt7a is a Wnt signaling pathway regulatory protein, DKK1 is a Wnt signaling pathway antagonist; TGF-β is a factor that promotes hair follicle cell apoptosis; TNFα and IL-6 are inflammatory factors; FABP3 is a fatty acid binding protein, and PPARγ promotes adipose differentiation. In the figure, each group of bar charts represents Control, DHT, DHT+G1, DHT+G2, DHT+G3, DHT+G4, DHT+G5, and DHT+G6 respectively.
[0073] Fig.14 The G7-G9 sequence peptides alleviated the changes of cell-related molecular markers after the androgen / seborrheic model was induced by DHT treatment using human primary dermal hair papilla cells in vitro. Among them, AR is the androgen receptor, 5α-reductase promotes the conversion of testosterone into dihydrotestosterone DHT, 3βHSD and 17βHSD are hormone synthesis-related enzymes; Caspase-3, Bax, Bcl-2 are markers of hair follicle cell apoptosis; CD200 and CD133 are markers of hair follicle stem cells, MMP-9 is a marker of hair follicle degeneration; Wnt7a is a Wnt signaling pathway regulatory protein, DKK1 is a Wnt signaling pathway antagonist; TGF-β is a factor that promotes hair follicle cell apoptosis; TNFα and IL-6 are inflammatory factors; FABP3 is a fatty acid binding protein, and PPARγ promotes adipose differentiation. In the figure, each group of bar graphs represents control, DHT, DHT+G7, DHT+G8, and DHT+G9, respectively.
[0074] Fig.15The G1-G6 sequence peptides alleviated the changes of cell-related molecular markers after the stress model was induced by cortisol treatment in vitro using human primary dermal hair papilla cells. Among them, GR is the glucocorticoid receptor, AR is the androgen receptor, Cyp21 is the gene for the enzyme related to cortisol production; STIP1 is the phosphorylated protein triggered by stress, GAS is a hair growth marker, CD200 is a hair follicle stem cell marker; Caspase-3 and Bcl-2 are markers of hair follicle cell apoptosis; Wnt7a is a Wnt signaling pathway regulatory protein, and DKK1 is a Wnt signaling pathway antagonist; TGF-β is a factor that promotes hair follicle cell apoptosis; TNF and IL-6 are inflammatory factors. In the figure, each group of bar graphs represents control, Cortisol, Cortisol+G1, Cortisol+G2, Cortisol+G3, Cortisol+G4, Cortisol+G5, and Cortisol+G6.
[0075] Fig.16 The G7-G9 sequence peptides alleviated the changes of cell-related molecular markers after the stress model was induced by cortisol treatment using human primary dermal hair papilla cells in vitro. Among them, GR is the glucocorticoid receptor, AR is the androgen receptor, Cyp21 is the gene for the enzyme related to cortisol production; STIP1 is a phosphorylated protein triggered by stress, GAS is a hair growth marker, CD200 is a hair follicle stem cell marker, MMP-9 is a hair follicle degeneration marker; Caspase-3 and Bcl-2 are markers of hair follicle cell apoptosis; Wnt7a is a Wnt signaling pathway regulatory protein, and DKK1 is a Wnt signaling pathway antagonist; TGF-β is a factor that promotes hair follicle cell apoptosis; TNF and IL-6 are inflammatory factors. In the figure, each group of bar graphs represents control, Cortisol, Cortisol+G7, Cortisol+G8, and Cortisol+G9, respectively. DETAILED DESCRIPTION
[0076] In order to better understand the content of the present invention, the content of the present invention is further explained below in conjunction with specific implementation cases, but the protection content of the present invention is not limited to the following embodiments.
[0077] the term
[0078] Hair follicle decline and / or hair loss caused by stress refers to the release of stress hormones, namely corticosteroids, by the adrenal glands due to chronic stress, which can prolong the resting phase of hair follicles, affect the activation of hair follicle stem cells, and cause hair regeneration disorders.
[0079] Seborrheic follicle decline and / or hair loss refers to the fact that androgens promote the sebaceous glands to secrete excessive oil, which accumulates around the hair follicles, causing the hair follicles to become smaller and ultimately leading to hair loss.
[0080] Stress-induced acne and / or dermatitis refers to the fact that stress acts on the hypothalamic-pituitary-adrenal axis, promoting the release of stress hormones and pro-inflammatory mediators, stimulating the sebaceous glands and hair follicles, and leading to increased skin inflammation.
[0081] Seborrheic acne and / or dermatitis refers to acne and / or dermatitis caused by excessive accumulation of oil in the hair follicles, which triggers an inflammatory response.
[0082] The polypeptide of the present invention is selected from the polypeptide sequence shown in any one of the following SEQ ID NOs. 1-9:
[0083]
[0084] The polypeptide-derived peptide is a polypeptide comprising the above sequence, or a sequence having a homology higher than 50% with the above sequence;
[0085] Derivatives of polypeptides and polypeptide-derived peptides are products obtained by conventional modification of the amino acid side chain groups, amino terminus or carboxyl terminus of the above-mentioned polypeptides or polypeptide-derived peptides, or products obtained by connecting tags for polypeptide or protein detection or purification, or products obtained by isotope labeling modification.
[0086] Example 1: Preparation of polypeptide sequences
[0087] The peptide sequence was synthesized by artificial synthesis.
[0088] The polypeptide sequence is as follows:
[0089]
[0090]
[0091] The above polypeptides are synthesized by conventional solid phase synthesis or liquid phase synthesis methods. The solid phase peptide synthesis method is to react from the amino acid at the C-terminus to the N-terminus, and then complete the amino acid linkage one by one through the steps of resin activation, amino acid linkage, elution protection, detection, etc., and then use excess ether precipitation and centrifugation, and the crude peptide is purified by HPLC, mass spectrometry analysis, and freeze-dried in liquid nitrogen for use.
[0092] Example 2: Peptide binding to receptor
[0093] The amino acid sequence identity of human and mouse GPR1 is 80%. The applicant has successfully constructed a cell line overexpressing human GPR1 and mouse GPR1 in HEK293 cells, co-cultured the cells with a fluorescently labeled polypeptide (Rhodamine-labeled LRH-G3), and observed the binding of the polypeptide to cells overexpressing human GPR1 and mouse GPR1 by immunofluorescence staining.
[0094] The experimental results are shown in Figure 1 Molecular docking simulation showed that LRH-G3 could bind to the extracellular domain of human GPR1 and mouse GPR1 (Figure A). The cell lines overexpressing human GPR1 and mouse GPR1 were successfully constructed by qPCR identification (Figure B). Fluorescence staining results showed that LRH-G3 could bind to HEK293-hGPR1 and HEK293-mGPR1 cells (Figure C).
[0095] Example 3: Peptides alleviate the inhibitory effect of chemerin on the cAMP signaling pathway by interacting with receptors
[0096] (1) Cyclic adenosine monophosphate (cAMP) ELISA:
[0097] ① Cell plating: 293T cells with high expression of GPR1 (293T GPR1 + / + ), 5 x 10 5 Each well was inoculated with 1 mL of culture medium. After 24 h of incubation in an incubator, the cells were starved overnight and different concentrations of LRH-G3 and LRH-G5 peptides (3 μM, 0.3 μM, 0.03 μM), Fosklin (25 μM) and chemerin (30 nM) were added for 6 h.
[0098] ②Sample preparation: Add 300 μL of cell lysis buffer to each well, place at 4°C for 20 minutes, scrape and collect cells with a cell scraper, mix by inverting, centrifuge at 12,000 rpm for 10 minutes, and collect the supernatant;
[0099] ③ Sample concentration determination: Sample concentration was determined by BCA method;
[0100] ④Cyclic adenosine monophosphate (cAMP) enzyme-linked immunosorbent assay:
[0101] a. Prepare the required reagents and prepare 3 replicate wells for each sample;
[0102] b, add 50 μL of sample or standard to a 96-well plate coated with antibodies; then add 25 μL of cAMP peroxidase tracer conjugate to each well;
[0103] c, add 50 μL Anti-cAMP antibody to each well and incubate slowly on a shaker for 30 minutes at room temperature;
[0104] d, wash five times with elution buffer, add 100 μL of chemical reflector to each well, and incubate at room temperature for 5 minutes;
[0105] e. Read the plate with an ELISA reader and record the luminescence value.
[0106] Figure 2 A shows that in 293T cells that highly express GPR1 (293T CMKLR1 + / + ) added with Forskolin can stimulate the cAMP level, while chemerin can inhibit the effect of Forskolin on cAMP. Adding different concentrations of LRH-G3 and LRH-G5 (3μM, 0.3μM, 0.03μM) can inhibit the effect of chemerin.
[0107] In summary, it can be inferred that LRH-G3 and LRH-G5 peptides can further inhibit the effect of chemerin on reducing cAMP concentration by specifically interacting with GPR1.
[0108] Example 4: Peptides can effectively inhibit the calcium (Ca) induced by chemerin 2+ ) Inflow
[0109] ① Cell plating: Wild-type 293T cells and 293T cells with high expression of GPR1 (293T GPR1 + / + ), 5 x 10 3 Each well was inoculated into a 96-well cell culture plate, with a culture medium volume of 200 μL per well, and placed in an incubator for 24 h before starvation overnight;
[0110] ② Reagent preparation: Dissolve probenecid in 1 mL of buffer solution to prepare a probenecid concentration of 250 nM. After shaking, add it to the fluorescent reagent and set aside for use;
[0111] ③ Remove the cell culture medium, add different concentration gradients (30 μM, 3 μM, 0.3 μM, 0.03 μM, 0.003 μM) of LRH-G3 and LRH-G5 peptides and chemerin (0.3 nM) for 30 minutes, and then add 100 μL of the above fluorescent reagent to each well;
[0112] ④ Place at 37℃ for 30 minutes, then place at room temperature for 30 minutes;
[0113] ⑤Measure the fluorescence absorbance at 494nm of excitation light and 516nm of emission light.
[0114] Figure 2 B shows that in 293T cells that highly express GPR1 (293T GPR1 + / + ) in the presence of chemerin at a concentration of 0.3 nM, which can promote the calcium 2+ ) flow signaling pathway, increasing calcium ions (Ca 2+ ) concentration. After adding different concentrations of LRH-G3 and LRH-G5 peptides, the calcium ion (Ca 2+ ) concentration, inhibiting the action of chemerin on calcium (Ca 2+ ) signaling pathway. However, in wild-type 293T cells that do not express GPR1, chemerin does not activate the calcium (Ca 2+ ) signaling pathway, LRH-G3 and LRH-G5 peptides have no effect on calcium (Ca 2+ ) signaling pathway, it can be inferred from this experiment that chemerin can activate calcium (Ca) by binding to the receptor GPR1 2+ ) signaling pathway, while LRH-G3 and LRH-G5 peptides can specifically inhibit the chemerin / GPR1 signaling pathway to reduce calcium ions (Ca 2+ )concentration.
[0115] Example 5: Expression and distribution of receptors of polypeptides and their derivatives
[0116] The human Protein Atlas was used to search for the expression and localization of the receptors of peptides and their derivatives in human brain, testis and ovarian tissue cells.
[0117] Search results are available at Figure 3 Through single-cell sequencing results, it can be seen that chemerin and its receptor GPR1 are expressed in various cells and neurons in human brain, testis and ovarian tissues.
[0118] Example 6: Detection of the ability of Cy5-labeled peptides to target brain, adrenal glands, ovaries and skin tissues of female mice
[0119] Adult female mice were injected with Cy5-labeled scrambled unrelated peptides, Cy5-labeled G3 and G5 peptides through the tail vein, 100 μL per mouse, and detected under a small animal imager 24 hours later.
[0120] The experimental results are shown in Figure 4 By comparing the results, it can be seen that the brain tissue, ovary, adrenal gland and skin tissue have obvious fluorescent signals compared with the control group, indicating that the G3 and G5 polypeptides have specific positioning and binding capabilities in the brain, ovary, adrenal gland and skin tissues.
[0121] Example 7: Detection of the ability of Cy5-labeled peptides to target brain, adrenal glands and skin tissues in male mice
[0122] Adult male mice were injected with Cy5-labeled scrambled unrelated peptides, Cy5-labeled G3 and G5 peptides through the tail vein, 100 μL per mouse, and detected under a small animal imager 24 hours later.
[0123] The experimental results are shown in Figure 5 By comparing the results, it can be seen that the brain, adrenal glands, testicles, epididymis and skin tissues all have obvious fluorescent signals compared with the control group, indicating that the polypeptide has specific positioning and binding capabilities in male brain, adrenal glands, gonads and skin tissues.
[0124] In summary, the results of Examples 6 and 7 demonstrate that the polypeptide can target the brain, adrenal glands, gonads and skin tissues of female and male mice in vivo. For the stress-induced alopecia model and the androgenic alopecia model, the brain (cerebral cortical neurons) and adrenal glands (cortical hormones) are related to stress regulation, and the male and female gonads are related to hormone regulation. The polypeptide may regulate the hair loss process by acting on target tissues regulated by stress and hormones.
[0125] Example 8: Expression and localization of receptor GPR1 in mouse hypothalamus
[0126] Adult C57 female mice were perfused with 4% paraformaldehyde, and then the hypothalamic tissues were dissected and isolated. After frozen sections, immunofluorescence staining was performed using GPR1, CRF and GnRH antibodies.
[0127] The experimental results are shown in Figure 6 It can be seen that GPR1 is specifically expressed in the hypothalamus and co-localizes with corticotropin-releasing factor (CRF) and gonadotropin-releasing hormone (GnRH).
[0128] Example 9: Expression and localization of receptor GPR1 in mouse pituitary
[0129] Adult C57 female mice were perfused with 4% paraformaldehyde, and then the pituitary tissues were dissected and separated. After frozen sections, immunofluorescence staining was performed using GPR1, FSH / LH, GH, TSH, and ACTH antibodies.
[0130] The experimental results are shown in Figure 7 It can be seen that GPR1 is specifically expressed in the pituitary gland and co-localized with pituitary hormones (follicle-stimulating hormone FSH, luteinizing hormone LH, growth hormone GH, thyroid-stimulating hormone TSH, and adrenocorticotropic hormone ACTH).
[0131] Example 10: Mouse stress-induced hair loss model
[0132] The stress-induced alopecia model was established in 7-week-old male C57BL / 6 mice using the steroid corticosteroid dexamethasone (modeling period 23 days), and the treatment group was topically applied with polypeptide G5 on the skin at the same time as the modeling.
[0133] The hair in the resting phase of the back was removed and depilatory cream was used. The model group mice were topically smeared with dexamethasone solution (100nM, 0.1mL, dissolved in propylene glycol) on the depilatory area once a day for 23 consecutive days. The treatment group mice were gavaged with peptide at a concentration of 0.5mg / kg / day while applying the drug every day.
[0134] The experimental results are shown in Figure 8 ,Through the comparison results, it can be seen that G polypeptide can alleviate the hair loss phenomenon in mice induced by local skin steroid corticosteroid dexamethasone.
[0135] Example 11: Mouse androgenic (seborrheic) alopecia model
[0136] 7-week-old C57BL / 6 mice were treated with DHT (modeling and experimental period totaled 28 days)
[0137] Remove 2*2cm of hair in the resting phase on the back and use a depilatory cream to remove hair. DHT solution (0.2%, w / v) was prepared in ethanol solution (50%, v / v). DHT solution (0.1mL / cm 2 ) for 28 consecutive days, while another group of mice was topically applied with peptide G5 at a concentration of 1 mg / kg / day while DHT was applied daily.
[0138] The experimental results are shown in Fig. 9 (female mice) and Fig.10 (Male mice). Through comparison results, it can be seen that G polypeptide can alleviate the hair loss phenomenon in mice caused by androgen.
[0139] Example 12: GPR1KO mouse androgenic (seborrheic) alopecia model
[0140] The androgenic alopecia model was established in wild-type male mice and GPR1KO male mice, and the modeling method was the same as Example 11.
[0141] The experimental results are shown in Fig.11 By comparing the modeling process of the two types of mice, it can be seen that on the 14th day of modeling, the degree of relief of androgen-induced hair loss on the back of GPR1KO mice was significantly better than that of wild-type mice.
[0142] Example 13: Detection of skin-related gene expression in wild-type and GPR1KO male mice on day 14 of DHT modeling
[0143] The results of related gene detection on the neck and back skin of wild-type mice and GPR1KO mice on the 14th day of modeling in Example 12 ( Fig.12 ) It can be seen that after 14 days of DHT modeling, the gene expressions of androgen receptor AR, 5α-reductase (promoting the conversion of testosterone into dihydrotestosterone DHT), testosterone production-related enzyme genes 3βHSD and 17βHSD, hair follicle cell apoptosis factor TGFβ, inflammatory factors TNFα and IL-6, hair follicle degeneration marker MMP-9, Wnt signaling pathway antagonist DKK1, adipose differentiation factor PPARγ and fatty acid binding protein FABP3 in the skin of male mice were significantly increased, and the expression of Wnt signaling pathway-related factor Wnt7a was significantly reduced. The change trend of related genes in GPR1KO male mice was significantly alleviated at 14 days of DHT modeling.
[0144] Example 14: DHT-induced androgen / seborrheic alopecia cell model
[0145] After removing serum-starved cells from primary human dermal papilla cells for 24 hours, 10 nM DHT was added to the culture medium for high androgen treatment, and the expression of cell molecular markers (5α-dehydrogenase, AR, CD200, IL-6, BCL2, BAX, FABP3, etc.) was detected 24 hours later. At the same time, different sequence peptides (G1-G9) were added for co-culture, and the expression of cell molecular markers was detected 24 hours later.
[0146] like Fig.13 and 14 The results showed that after DHT treatment, the expression of cellular androgen receptor AR, 5α reductase and androgen synthesis-related enzymes (3βHSD, 17βHSD) increased significantly, while hair follicle cell apoptosis (Caspase-3, BCL2, BAX) increased, the expression of hair follicle cell apoptosis factor TGF-β and inflammatory factors (TNF-α, IL-6) increased, the expression of hair follicle degeneration marker MMP-9 increased, the expression of hair follicle stem cell markers CD133 and CD200 decreased, the expression of fatty acid binding protein FABP3 and adipose differentiation factor PPARγ increased, the expression of Wnt signaling pathway antagonist DKK1 increased, and the expression of Wnt signaling pathway-related factor Wnt7a significantly decreased. G1-G9 sequence peptides can alleviate these changes to a certain extent.
[0147] It is shown that the polypeptide of the present invention can be used to alleviate androgenic alopecia, especially androgenic / seborrheic follicle decline or alopecia, and androgenic / seborrheic acne or dermatitis caused by significantly increased expression of androgen receptor AR5α reductase and androgen synthesis-related enzymes (3βHSD, 17βHSD3), hair follicle cell apoptosis, abnormally increased expression of inflammatory factors, and hair follicle degeneration.
[0148] Example 15: Cortisol-induced stress-induced hair loss cell model
[0149] After removing serum-starved cells for 24 hours, human primary dermal papilla cells were cultured in a culture medium containing 1 μM cortisol for 24 hours. The expression of cell molecular markers (5α-dehydrogenase, GR, AR, CD200, IL-6, BCL2, BAX, etc.) was detected after 24 hours. At the same time, different sequence peptides (G1-G9) were added for co-culture, and the expression of cell molecular markers was detected after 24 hours.
[0150] like Fig.15 and 16 The results showed that after cortisol treatment, the expression of cellular androgen receptor AR, glucocorticoid receptor GR and cortisol production-related enzyme Cyp21 increased significantly, while hair follicle cell apoptosis (Caspase-3, Bcl2) increased, pressure-induced phosphorylated protein STIP1 and hair follicle apoptosis-promoting factor TGFβ, inflammatory factors (TNF, IL-6) increased, hair follicle degeneration marker MMP-9 increased, hair growth marker GAS and hair follicle stem cell marker CD200 decreased, Wnt signaling pathway antagonist DKK1 increased, and Wnt signaling pathway-related factor Wnt7a expression significantly decreased. G1-G9 sequence peptides can alleviate these changes to a certain extent.
[0151] It is shown that the polypeptide of the present invention can be used to alleviate stress-induced hair loss, especially hair follicle decline or hair loss caused by stress due to hair follicle cell apoptosis, abnormally increased expression of phosphorylated protein STIP1, abnormally increased expression of inflammatory factors, and reduced hair follicle growth, as well as acne or dermatitis caused by stress.
[0152] In summary, the polypeptide and its related substances provided by the present invention can be used to prepare therapeutic drugs for combating hair follicle decline and / or hair loss and other related diseases caused by stress, and / or androgen, and / or seborrheic dermatitis.
[0153] The above description is only a specific implementation mode of the present invention, not all implementation modes. Any equivalent changes made to the technical solution of the present invention by ordinary technicians in this field after reading the specification of the present invention are 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 drug for treating hair follicle decline and / or alopecia caused by stress, seborrheic hair follicle decline and / or alopecia, acne and / or dermatitis caused by stress, and seborrheic acne and / or dermatitis; The polypeptide is selected from the polypeptide sequence shown in any one of the following SEQ ID NO.1-9: The polypeptide-derived peptide is a polypeptide comprising the above sequence, or a sequence having a homology higher than 50% with the above sequence; The derivative is a product obtained by conventionally modifying the amino acid side chain groups, the amino terminus or the carboxyl terminus of the polypeptide or polypeptide-derived peptide, or a product obtained by connecting a tag for polypeptide or protein detection or purification, or a product obtained by isotope labeling modification; Preferably, the product obtained by conventional modification is selected from the product obtained by fluorescent group modification, or the product obtained by phosphorylation modification, or the product obtained by disulfide bond-based cyclization modification, or the product obtained by biotin labeling modification, or the product obtained by photosensitizer modification, or the product obtained by azide modification, or the product obtained by PEG modification, or the product obtained by methylation modification, or the product obtained by fluorescence quenching group modification, or the product obtained by protein coupling modification, or the product obtained by small molecule compound modification, or the product obtained by amino modification, amidation modification, hydroxylation modification, carboxylation modification, carbonylation modification, alkylation modification, acetylation modification, esterification modification, glycosylation modification; Preferably, the modification of the amino terminus is selected from amino terminus acetylation modification; Preferably, the modification of the carboxyl terminus is selected from carboxyl terminus acylation modification; Preferably, the modification of the side chain 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 modification can be used to prepare a photosensitizer; 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.
2. The use according to claim 1, characterized in that The hair follicle decline and / or hair loss caused by stress is hair follicle decline and / or hair loss caused by stress in male or female subjects; The seborrheic follicle decline and / or hair loss is seborrheic follicle decline and / or hair loss of a male or female subject; The acne and / or dermatitis caused by stress is acne and / or dermatitis caused by stress in a male or female subject; The seborrheic acne and / or dermatitis is seborrheic acne and / or dermatitis of a male or female subject.
3. Use of a substance capable of binding to and exerting its function on a GPR1 receptor in the preparation of a medicament for treating and / or preventing hair follicle decline and / or hair loss caused by stress, seborrheic hair follicle decline and / or hair loss, acne and / or dermatitis caused by stress, and seborrheic acne and / or dermatitis; Preferably, the substance capable of binding to the GPR1 receptor and exerting its function is a substance capable of binding to the GPR1 receptor and triggering its downstream signaling pathway; Preferably, the substance capable of binding to the GPR1 receptor and exerting its function is selected from inhibitory enzymes, receptor-binding polypeptides, antibodies, siRNAs, shRNAs, antisense RNA and DNA enzymes or a combination thereof, and expression vectors containing siRNAs, shRNAs and antisense molecules.
4. The use according to claim 3, characterized in that The substance capable of binding to the GPR1 receptor and exerting its function is a polypeptide, a polypeptide-derived peptide, or a derivative of a polypeptide and a polypeptide-derived peptide, The polypeptide is selected from the polypeptide sequence shown in any one of the following SEQ ID NO.1-9: The polypeptide-derived peptide is a polypeptide comprising the above sequence, or a sequence having a homology higher than 50% with the above sequence; The derivative is a product obtained by conventionally modifying the amino acid side chain groups, the amino terminus or the carboxyl terminus of the polypeptide or polypeptide-derived peptide, or a product obtained by connecting a tag for polypeptide or protein detection or purification, or a product obtained by isotope labeling modification; Preferably, the product obtained by conventional modification is selected from the product obtained by fluorescent group modification, or the product obtained by phosphorylation modification, or the product obtained by disulfide bond-based cyclization modification, or the product obtained by biotin labeling modification, or the product obtained by photosensitizer modification, or the product obtained by azide modification, or the product obtained by PEG modification, or the product obtained by methylation modification, or the product obtained by fluorescence quenching group modification, or the product obtained by protein coupling modification, or the product obtained by small molecule compound modification, or the product obtained by amino modification, amidation modification, hydroxylation modification, carboxylation modification, carbonylation modification, alkylation modification, acetylation modification, esterification modification, glycosylation modification; Preferably, the modification of the amino terminus is selected from amino terminus acetylation modification; Preferably, the modification of the carboxyl terminus is selected from amidation modification of the carboxyl terminus; Preferably, the modification of the side chain 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 modification can be used to prepare a photosensitizer; 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.
5. The use according to claim 3, characterized in that: The hair follicle decline and / or hair loss caused by stress is hair follicle decline and / or hair loss caused by stress in male or female subjects; The seborrheic follicle decline and / or hair loss is seborrheic follicle decline and / or hair loss of a male or female subject; The acne and / or dermatitis caused by stress is acne and / or dermatitis caused by stress in a male or female subject; The seborrheic acne and / or dermatitis is seborrheic acne and / or dermatitis of a male or female subject.
6. Use of a polypeptide, a polypeptide-derived peptide, a polypeptide and a polypeptide-derived peptide derivative, or a substance capable of binding to and functioning with a GPR1 receptor 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, and dermatitis; The polypeptide is selected from the polypeptide sequence shown in any one of the following SEQ ID NO.1-9: The polypeptide-derived peptide is a polypeptide comprising the above sequence, or a sequence having a homology higher than 50% with the above sequence; The derivative is a product obtained by conventionally modifying the amino acid side chain groups, the amino terminus or the carboxyl terminus of the polypeptide or polypeptide-derived peptide, or a product obtained by connecting a tag for polypeptide or protein detection or purification, or a product obtained by isotope labeling modification; Preferably, the product obtained by conventional modification is selected from the product obtained by fluorescent group modification, or the product obtained by phosphorylation modification, or the product obtained by disulfide bond-based cyclization modification, or the product obtained by biotin labeling modification, or the product obtained by photosensitizer modification, or the product obtained by azide modification, or the product obtained by PEG modification, or the product obtained by methylation modification, or the product obtained by fluorescence quenching group modification, or the product obtained by protein coupling modification, or the product obtained by small molecule compound modification, or the product obtained by amino modification, amidation modification, hydroxylation modification, carboxylation modification, carbonylation modification, alkylation modification, acetylation modification, esterification modification, glycosylation modification; Preferably, the modification of the amino terminus is selected from amino terminus acetylation modification; Preferably, the modification of the carboxyl terminus is selected from amidation modification of the carboxyl terminus; Preferably, the modification of the side chain 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 modification can be used to prepare a photosensitizer; 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; Preferably, the hair loss includes stress-induced hair follicle decline and / or hair loss in males and females, androgenic 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 stress acne, androgenic acne; Preferably, the dermatitis includes male and female stress dermatitis and androgenic dermatitis.
7. Use of a substance capable of binding to a GPR1 receptor in the preparation of a detection reagent targeting the brain, skin, adrenal glands, and gonadal system; Preferably, the brain is a female or male brain; Preferably, the skin is female or male skin; Preferably, the adrenal gland is a female or male adrenal gland; Preferably, the male gonadal system is the testicles and epididymis; Preferably, the female gonadal system is the ovary.
8. The use according to claim 7, characterized in that The substance capable of binding to the GPR1 receptor is a polypeptide, a polypeptide-derived peptide, or a derivative of a polypeptide and a polypeptide-derived peptide; Preferably, the polypeptide is selected from the polypeptide sequence shown in any one of the following SEQ ID NOs. 1-9: The polypeptide-derived peptide is a polypeptide comprising the above sequence, or a sequence having a homology higher than 50% with the above sequence; The derivative is a product obtained by conventionally modifying the amino acid side chain groups, the amino terminus or the carboxyl terminus of the polypeptide or polypeptide-derived peptide, or a product obtained by connecting a tag for polypeptide or protein detection or purification, or a product obtained by isotope labeling modification; Preferably, the product obtained by conventional modification is selected from the product obtained by fluorescent group modification, or the product obtained by phosphorylation modification, or the product obtained by disulfide bond-based cyclization modification, or the product obtained by biotin labeling modification, or the product obtained by photosensitizer modification, or the product obtained by azide modification, or the product obtained by PEG modification, or the product obtained by methylation modification, or the product obtained by fluorescence quenching group modification, or the product obtained by protein coupling modification, or the product obtained by small molecule compound modification, or the product obtained by amino modification, amidation modification, hydroxylation modification, carboxylation modification, carbonylation modification, alkylation modification, acetylation modification, esterification modification, glycosylation modification; Preferably, the modification of the amino terminus is selected from amino terminus acetylation modification; Preferably, the modification of the carboxyl terminus is selected from amidation modification of the carboxyl terminus; Preferably, the modification of the side chain 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 modification can be used to prepare a photosensitizer; 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.
9. Use of a substance capable of binding to a GPR1 receptor in the preparation of medicines, medical and cosmetic products, cosmetics, health products, foods, and additives for inhibiting the increase of corticosteroids, or inhibiting the inflammatory response and / or hair follicle decline and / or hair loss caused by the increase of corticosteroids; Preferably, the substance capable of binding to the GPR1 receptor is a polypeptide, a polypeptide-derived peptide, or a derivative of a polypeptide and a polypeptide-derived peptide; Preferably, the polypeptide is selected from the polypeptide sequence shown in any one of the following SEQ ID NOs. 1-9: The polypeptide-derived peptide is a polypeptide comprising the above sequence, or a sequence having a homology higher than 50% with the above sequence; The derivative is a product obtained by conventionally modifying the amino acid side chain groups, the amino terminus or the carboxyl terminus of the polypeptide or polypeptide-derived peptide, or a product obtained by connecting a tag for polypeptide or protein detection or purification, or a product obtained by isotope labeling modification; Preferably, the product obtained by conventional modification is selected from the product obtained by fluorescent group modification, or the product obtained by phosphorylation modification, or the product obtained by disulfide bond-based cyclization modification, or the product obtained by biotin labeling modification, or the product obtained by photosensitizer modification, or the product obtained by azide modification, or the product obtained by PEG modification, or the product obtained by methylation modification, or the product obtained by fluorescence quenching group modification, or the product obtained by protein coupling modification, or the product obtained by small molecule compound modification, or the product obtained by amino modification, amidation modification, hydroxylation modification, carboxylation modification, carbonylation modification, alkylation modification, acetylation modification, esterification modification, glycosylation modification; Preferably, the modification of the amino terminus is selected from amino terminus acetylation modification; Preferably, the modification of the carboxyl terminus is selected from amidation modification of the carboxyl terminus; Preferably, the modification of the side chain 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 modification can be used to prepare a photosensitizer; 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.
10. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises a therapeutically effective amount of a polypeptide, a polypeptide-derived peptide, or a polypeptide and a polypeptide-derived peptide derivative, and / or a substance capable of binding to a GPR1 receptor and exerting a function; The polypeptide is selected from the polypeptide sequence shown in any one of the following SEQ ID NO.1-9: The polypeptide-derived peptide is a polypeptide comprising the above sequence, or a sequence having a homology higher than 50% with the above sequence; The derivative is a product obtained by conventionally modifying the amino acid side chain groups, the amino terminus or the carboxyl terminus of the polypeptide or polypeptide-derived peptide, or a product obtained by connecting a tag for polypeptide or protein detection or purification, or a product obtained by isotope labeling modification; Preferably, the product obtained by conventional modification is selected from the product obtained by fluorescent group modification, or the product obtained by phosphorylation modification, or the product obtained by disulfide bond-based cyclization modification, or the product obtained by biotin labeling modification, or the product obtained by photosensitizer modification, or the product obtained by azide modification, or the product obtained by PEG modification, or the product obtained by methylation modification, or the product obtained by fluorescence quenching group modification, or the product obtained by protein coupling modification, or the product obtained by small molecule compound modification, or the product obtained by amino modification, amidation modification, hydroxylation modification, carboxylation modification, carbonylation modification, alkylation modification, acetylation modification, esterification modification, glycosylation modification; Preferably, the modification of the amino terminus is selected from amino terminus acetylation modification; Preferably, the modification of the carboxyl terminus is selected from amidation modification of the carboxyl terminus; Preferably, the modification of the side chain 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 modification can be used to prepare a photosensitizer; 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; Preferably, the pharmaceutical composition further contains one or more pharmaceutically acceptable carriers; Preferably, the carrier is selected from diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, adsorption carriers, surfactants or lubricants; Preferably, the pharmaceutical composition is prepared into tablets, granules, capsules, oral liquids, inhalation liquids, smears, sprays, drops, microneedles or injections; Preferably, the pharmaceutical composition is administered to a subject orally, topically, as a suppository, intravenously, intraperitoneally, intramuscularly, intralesionally, intrathecally, intranasally or subcutaneously; Preferably, the administration routes of the pharmaceutical composition include parenteral administration such as intravenous, intramuscular, intraarterial, intradermal, subcutaneous, intraperitoneal, intraventricular and intracranial; transmucosal administration such as buccal, sublingual, palatal, transgingival, nasal, vaginal, rectal or transdermal, and other delivery modes include but are not limited to the use of liposome preparations, intravenous infusion, and transdermal patches.