Application of Ectoine as enorphin generation accelerant

Abstract

The invention provides application of Ectoin as an enkephalin generation accelerant, and relates to the technical field of cosmetics. According to the present invention, it is found for the first time that Ectoine can promote the generation of beta-enorphin in skin cells, such that Ectoine can be used as a nerve beautifying component, for example, the Ectoine can improve the pain threshold of skin, and has a positive alleviation effect on skin uncomfortable reactions such as aching feeling, itching feeling and the like. Furthermore, based on the promotion effect of Ectoine on the generation of beta-enorphin, the Ectoine can play multiple skin care effects in multiple aspects, including promoting skin barrier repair, relieving skin sensitivity, promoting skin wound healing, accelerating skin regeneration, maintaining skin steady state, relieving inflammatory response and the like, so that the skin care scene of Ectoine is expanded, and the skin care effect of Ectoine is improved. And a new application is developed for Ectoin.

Description

Technical Field

[0001] The present application relates to the field of neurocosmetic technology, and in particular to the application of ectoine as an endorphin production promoter. Background Art

[0002] Neurocosmetics is an innovative product developed recently in response to research progress on skin aging, which has brought revolutionary changes to the cosmetics industry. The concept of "neurocosmetics" was first proposed by Professor Misery in 2000, and then received great attention at the New York Society of Cosmetic Chemists in 2007. It is a non-toxic, bioactive topical product whose ingredients work at the neural level, that is, achieving neurocosmetics.

[0003] In order to improve the appearance and defects of the skin, current research on neurocosmetics is mainly focused on ingredients with similar structures to endogenous neurotransmitters. Neurocosmetic ingredients can exert their effects through the following mechanisms: (1) Directly acting on the endings of skin nerve fibers as regulators of neurotransmitter release. For example, botulinum toxin-like peptides can promote facial muscle relaxation to smooth wrinkles. In another example, neuron-inhibiting peptides are mainly used to reduce the reactivity of allergic skin to environmental stimuli (temperature changes, air humidity, smoke, etc.). (2) Acting as agonist / antagonist molecules of neuropeptide receptors or neurotransmitter effector modulators to regulate the function of non-neuronal cells.

[0004] The modern beauty industry is working on discovering new neurocosmetic functional ingredients to improve the interaction between the skin and the nervous system. Many cosmetic companies have begun formulating neurocosmetics that affect the skin's neurotransmitters through different mechanisms of action, thereby showing their activity on the skin's nervous system.

[0005] β-endorphin is an endogenous neurotransmitter peptide produced by the anterior pituitary gland and is a natural analgesic and pleasure hormone in the human body. β-endorphin receptors are expressed in skin cells, in which case β-endorphin has been shown to accelerate skin regeneration and wound healing. In human keratinocytes, the presence of the β-endorphin / μ-opioid receptor system has been shown to be associated with keratinocyte differentiation, wound healing processes, and migration (references: Carli, B. Feeling good about neuro-cosmetics. Househ. Pers. Care Today 2016, 11, 9-11.; Schmelz, M.; Paus, R. Opioids and the skin: “Itchy” perspectives beyond analgesia and abuse. J. Investig. Dermatol. 2007, 127, 1287-1289.; Bigliardi, PL; Tobin, DJ; Gaveriaux-Ruff, C.; Bigliardi-Qi, M. Opioids and the skin—Where do we stand? Exp.Dermatol.2009,18,424-430.; Bigliardi, PL; Dancik, Y.; Neumann, C.; Bigliardi-Qi, M. Opioids and skin homeostasis, regeneration and ageing—What's the evidence? Exp.Dermatol.2016,25,586-591.;Bigliardi-Qi,M.;Bigliardi,PL;Eberle,AN;Büchner,S.;Rufli,T.β-Endorphin Stimulates Cytokeratin 16Expression andDownregulatesμ-Opiate Receptor Expression in Human Epidermis. J. Investig. Dermatol. 2000, 114, 527-532.). β-endorphin upregulates the expression of transforming growth factor β type II receptor in human skin organ cultures, suggesting that β-endorphin plays a crucial role not only in pain control but also in wound healing (reference: PL Bigliardi, LT Sumanovski, S. T.Ru￡I,T.Ru￡I,DifferentExpression of l-Opiate Receptor in Chronic and Acute Wounds and the Eiect of b-Endorphin on Transforming Growth Factor b Type II Receptor and Cytokeratin16 Expression,THE JOURNAL OF INVESTIGATIVE DERMATOLOGY.2002,120,145-152.). PROVITAL found that Rhodiola rosea extract can stimulate the production of beta-endorphin in the skin, making the skin more radiant (References: Li, Y.; Pham, V.; Bui, M.; Song, L.; Wu, C.; Walia, A.; Uchio, E.; Smith-Liu, F.; Zi, X. Rhodiola rosea L.: An Herb with Anti-Stress, Anti-Aging, and Immunostimulating Properties for Cancer Chemoprevention. Curr. Pharmacol. Rep. 2017, 3, 384-395.; Talukdar, D.; Rhodiola Rosea, L. A critical review on biology, medicinal properties and pharmacological manifestations. J. Nat. Prod. Resour. 2015, 1, 4-9.; Dieamant, G.C.; Velazquez Pereda, MDC; Eberlin, S.; Nogueira, C.; Werka, RM; DeQueiroz, MLS Neuroi mmunomodulatory compound for sensitive skin care: Invitro and clinical ass essment. J. Cosmet. Dermatol. 2008, 7, 112-119.).β-endorphin reduces UVB-induced inflammatory responses by inactivating the NF-κB pathway and reverses UVB-induced skin damage, including the destruction of skin barrier function (reference: Hyung-Su Kim, Hyoung-June Kim, Yong Deog Hong, Eui DongSon, Si-Young Cho, β-endorphin suppresses ultraviolet Birradiation induced epidermal barrier damage by regulating inflammation-dependent mTORC1signaling, nature portfolio, 2023, 13: 22357). Immune cell-derived β-endorphin can activate peripheral opioid receptors on sensory neurons, thereby inhibiting pain in inflamed tissues. The transport of β-endorphin to damaged tissues is enhanced, which helps prevent excessive and / or chronic inflammatory pain (reference: Shaaban A. Mousa, Qin Zhang, Nicolle Sitte, Ru-Rong Ji, Christopher Stein. β-Endorphin-containing memory-cells and μ-opioid receptors undergo transport to peripheral inflamed tissue. JOURNAL OF NEUROIMMUNOLOGY, 2001. 115, 71-78.). It is also reported that β-endorphin has the effects of regulating itch and counteracting nociception (reference: Bigliardi, PL; Dancik, Y.; Neumann, C.; Bigliardi-Qi, M. Opioids and skin homeostasis, regeneration and ageing—What's the evidence? Exp. Dermatol. 2016, 25, 586-591.). Beta-endorphin has been shown to increase the skin’s pain threshold, meaning it can reduce the skin’s sensory sensitivity and may have a relieving effect on skin conditions that are painful or itchy, such as chronic skin diseases and inflammation.In addition, β-endorphin is involved in maintaining skin homeostasis by binding to μ-opioid receptors on the skin (references: Bigliardi, PL; Dancik, Y.; Neumann, C.; Bigliardi-Qi, M. Opioids and skin homeostasis, regeneration and ageing—What's the evidence? Exp. Dermatol. 2016, 25, 586-591.).

[0006] Therefore, the development of new neurocosmetic ingredients that can have beneficial effects on the skin remains a topic of active research in this field. Summary of the invention

[0007] The present application found that ectoine has the effect of promoting the production of endorphins in the skin, so it can be used as a neurocosmetic ingredient in skin care related to endorphin production.

[0008] On the one hand, the present application provides the use of ectoine as an endorphin production promoter.

[0009] Furthermore, the endorphin includes β-endorphin.

[0010] On the other hand, the present application also provides the use of ectoine in promoting the production of endorphins by skin cells.

[0011] Preferably, the skin cells include dermal fibroblasts and keratinocytes.

[0012] Preferably, ectoine is applied to the skin to produce endorphins in skin cells.

[0013] Furthermore, the endorphin includes β-endorphin.

[0014] This application proves for the first time that Ecdoin can promote the production of β-endorphin in skin cells, and can be used as an effective ingredient of endorphin production promoter, providing a new effective ingredient in the field of neurocosmetology.

[0015] On the other hand, the present application also provides the use of ectoine in the preparation of neurocosmetic products.

[0016] Furthermore, ectoine is used as an endorphin production promoter in the preparation of neurocosmetic products.

[0017] On the other hand, the present application also provides the use of ectoine in the preparation of a product for increasing the pain threshold of the skin.

[0018] Furthermore, ectoine is used as an endorphin production promoter in the preparation of products for increasing the pain threshold of the skin.

[0019] On the other hand, the present application also provides the use of ectoine in preparing a product for reducing and / or alleviating the sensory sensitivity of the skin.

[0020] Furthermore, ectoine is used as an endorphin production promoter in the preparation of products for reducing and / or alleviating the sensory sensitivity of the skin.

[0021] Furthermore, the product can reduce and / or relieve the pain or itching of the skin.

[0022] Furthermore, the endorphin includes β-endorphin.

[0023] Preferably, the application is achieved by promoting the production of β-endorphin and increasing the content of β-endorphin through ectoine.

[0024] Furthermore, the neurocosmetic product has one or more of the following effects: promoting skin barrier repair, soothing skin sensitivity, promoting skin wound healing, accelerating skin regeneration, maintaining skin homeostasis, and alleviating inflammatory response. More preferably, it has two, three, four, five, or six of the above effects.

[0025] Preferably, the application is achieved by promoting the production of β-endorphin and increasing the content of β-endorphin through ectoine.

[0026] On the other hand, the present application also provides an endorphin production promoter, which contains ectoine.

[0027] On the other hand, the present application also provides a neurocosmetic product, which contains ectoine.

[0028] The neurocosmetic product or endorphin production promoter described in the present application may also be added with excipients, which may be appropriate solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesive agents, integrators, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants and deflocculating agents, filter aids, release retardants, etc.

[0029] The neurocosmetic product or endorphin production promoter described in the present application can be prepared by a general method, wherein one or more diluents or carriers can be added, for example, in an external application form, such as an emulsion, a suspension, and the like.

[0030] On the other hand, the present application also provides the use of the neurocosmetic product in achieving at least two of the following effects: promoting skin barrier repair, soothing skin sensitivity, promoting skin wound healing, accelerating skin regeneration, maintaining skin homeostasis, and alleviating inflammatory response. More preferably, it has three, four, five, or six of the above effects.

[0031] The amount of ectoine to be used according to the present application can vary within a wide range to determine a suitable amount, depending on the desired cosmetic effect. For example, its content in the product can be 0.001%-5%, more preferably 0.01%-1%.

[0032] The present invention has the following beneficial effects:

[0033] The present invention discovers for the first time that ectoine can promote the production of β-endorphin in skin cells, so that ectoine can be used as a neurocosmetic ingredient. For example, it can increase the pain threshold of the skin and has a positive effect on alleviating skin discomfort reactions, such as pain, itching, etc.

[0034] Furthermore, based on the promoting effect of Ecdoine on the production of β-endorphin, it can play multiple skin care effects in many aspects, including promoting skin barrier repair, soothing skin sensitivity, promoting skin wound healing, accelerating skin regeneration, maintaining skin homeostasis, and alleviating inflammatory responses, etc., which expands the skin care scenarios of Ecdoine and develops new uses for Ecdoine. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] The drawings described herein are used to provide a further understanding of the present application and constitute a part of the present application. The illustrative embodiments of the present application and their descriptions are used to explain the present application and do not constitute an improper limitation on the present application. In the drawings:

[0036] Figure 1 These are fluorescent images of human dermal fibroblasts, including, from left to right, a photo of β-endorphin fluorescent staining, a photo of cell nuclei stained with DAPI, and a fusion photo of cell nuclei stained with β-endorphin fluorescent staining and DAPI staining;

[0037] Figure 2 This is a statistical diagram of the relative fluorescence intensity of β-endorphin. DETAILED DESCRIPTION

[0038] In order to more clearly explain the overall concept of the present application, the following is described in detail in conjunction with the accompanying drawings of the specification by way of embodiment. In the following description, a large number of specific details are provided to provide a more thorough understanding of the present invention. However, it is obvious to those skilled in the art that the present invention can be implemented without one or more of these details. In other examples, in order to avoid confusion with the present invention, some technical features well known in the art are not described.

[0039] Unless otherwise specified, in the following embodiments, the reagents or instruments used without indicating the manufacturer are all conventional products that can be purchased commercially.

[0040] Among them, the carbon dioxide incubator (160i) was purchased from Thermo, the biological safety cabinet was purchased from Thermo, and the fluorescence inverted microscope was purchased from Leica.

[0041] The cell models and experimental materials involved in the following examples are from the following Table 1:

[0042] Table 1

[0043] Cell models and experimental materials Suppliers Human dermal fibroblasts (NHDF) ATCC DMEM medium Gibco <![CDATA[Ectoin (Xi'anyan TM Ectoin)]]> Bloomage Biopharmaceuticals β-endorphin Primary antibody Invitrogen

[0044] Unless otherwise stated, the experimental methods, detection methods, and preparation methods disclosed in the present invention all adopt conventional techniques in the field of molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell culture, recombinant DNA technology, and related fields, and can be specifically carried out according to Molecular Cloning: A Laboratory Manual (Fourth Edition).

[0045] In the following examples, unless otherwise specified, % means wt %, i.e. weight percentage.

[0046] In addition, the "water" mentioned in the present invention includes any feasible water that can be used in the art, such as deionized water, distilled water, ion exchange water, double distilled water, high-purity water, and purified water.

[0047] Example 1 Test on the secretion of β-endorphin promoted by Ecodoxin

[0048] In this example, immunofluorescence was used to qualitatively and quantitatively determine the changes in the secretion level of β-endorphin in human dermal fibroblasts after treatment with icodoxin compared with the control group, and whether icodoxin could increase the secretion level of β-endorphin was evaluated based on the significant difference.

[0049] Specific experimental methods include:

[0050] 1. Preparation of experimental sample solution: dilute the icodone powder with DMEM culture medium to prepare a standby solution containing 0.1 wt% icodone.

[0051] 2. Treatment method: Human dermal fibroblast cell line was plated in a 24-well plate containing DMEM medium with cell slides at a density of 15,000 / well, and cultured in an incubator for 24 hours. The cells were grouped (as shown in Table 2) and treated with the following treatment conditions:

[0052] (1) Untreated group: The DMEM medium was aspirated and replaced with fresh DMEM medium, and the plate was cultured for 72 h.

[0053] (2) Experimental group: The DMEM medium was aspirated and replaced with the experimental sample solution, and the plate was cultured for a further 72 h.

[0054] 3. Immunofluorescence test: Collect cell slides separately, fix the cells with ice methanol for 15 minutes, add anti-β-endorphin antibody and incubate at 4°C overnight, discard the supernatant, add Goat-anti-Rb IgG secondary antibody and incubate at room temperature for 1.5 hours, and use DAPI mounting medium to seal the slides. Observe and take fluorescence images under a Leica inverted fluorescence microscope (the results are shown in Figure 1 ), and then the fluorescence intensity was quantified using Image J software. The fluorescence intensity of the wells in the untreated group was defined as N0 and set to 100% as a reference for the β-endorphin content under normal conditions. The fluorescence intensity of the experimental group was defined as Nt, and Nt / N0 was the relative fluorescence intensity between the treated group and the untreated group. GraphPad prism6 software was used for plotting, and the significance of the β-endorphin expression levels between samples was tested, *p<0.05 indicated a statistically significant difference. The grouping and detection conditions are shown in Table 2, and the fluorescence intensity analysis results are shown in Table 3. Table 3 plots the results. Figure 2 .

[0055] Table 2

[0056]

[0057] Table 3 Relative fluorescence intensity

[0058] Untreated group Experimental Group Corresponding fluorescence intensity 1±0.0133 1.17±0.0678 p - **

[0059] like Figure 1 , Figure 2 As shown in Table 3, compared with the untreated group, the expression level of β-endorphin in human dermal fibroblasts in the experimental group was significantly increased, which indicates that Ecdoin can increase the expression level of β-endorphin.

[0060] The raw material according to the embodiment of the present disclosure can be a raw material for a skin external preparation containing the active ingredient. More specifically, it can be a raw material for a skin coating formulation. In one embodiment, the coating site can include the skin, i.e., all tissues covering the body surface of the face or scalp, body, etc.

[0061] The above is only an embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various changes and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the scope of the claims of the present application.

Claims

1. The application of ectoine as an endorphin production promoter.

2. The application of Ecdoin in promoting the production of endorphins in skin cells.

3. Application of Ecdoin in the preparation of products that promote the production of endorphins.

4. Application of Ecdoin in the preparation of neurocosmetic products.

5. Application of Ecdoin in the preparation of products for increasing the pain threshold of the skin.

6. Use of Ekto in the preparation of products for reducing and / or alleviating the sensory sensitivity of the skin.

7. The use according to claim 5 or 6, characterized in that: The product can reduce and / or relieve the pain or itching sensation of the skin.

8. The use according to any one of claims 1 to 6, characterized in that: The endorphins include beta-endorphin.

9. The use according to any one of claims 3 to 6, characterized in that: The product also has one or more of the following effects: promoting skin barrier repair, soothing skin sensitivity, promoting skin wound healing, accelerating skin regeneration, maintaining skin homeostasis, and alleviating inflammatory responses.

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