Composition for reducing scalp grease and application thereof

Abstract

The invention discloses a composition for reducing scalp grease and application thereof. The invention discloses a functional composition which comprises an effective amount of betulin and glycyrrhizic acid, and the betulin and the glycyrrhizic acid achieve a remarkable effect of reducing scalp grease through mutual synergy and synergism. According to the technical scheme, a brand-new and efficient solution is provided for scalp grease regulation and control.

Description

Technical Field

[0001] This invention relates to the field of daily chemical products technology, and more specifically, to a composition for reducing scalp oil and its application. Background Technology

[0002] Excessive sebum secretion ( Seborrhea ) This can lead to excessive oil production on the skin, fostering the growth of pathogenic microorganisms and causing problems such as dandruff, seborrheic dermatitis, hair loss, and acne. Current mainstream oil-control strategies... ( Such as physical adsorption, surfactant cleaning, astringents, and limited pathway inhibitors. ) However, current methods often suffer from superficial effects, short-lived results, limited target selection, or potential side effects. Developing novel targeted interventions that address the underlying mechanisms of sebum synthesis is crucial to resolving unmet clinical needs.

[0003] terminally differentiated cells of sebaceous glands ( Sebocytes ) It's sebum. ( Sebum ) The main functional unit of synthesis, the sebum it secretes, contains four core components: triglycerides. ( Triglycerides, TAGs )( 40-60% ) wax esters ( Wax esters,WEs )( 20-30% ) squalene ( Squalene )( 10-15% ) and cholesterol esters ( Cholesteryl esters, CEs )( 3-6% ) .

[0004] The synthesis of these lipids is regulated by a highly complex signaling network, including androgen receptors. ( AR ) The insulin / IGF-1, PPARγ, and AMPK pathways form the core framework, while sterol regulatory element-binding protein 1... ( SREBP1 ) It is widely recognized as the central hub for regulating the network.

[0005] androgen receptor ( AR ) Pathway: Androgens ( Testosterone, dihydrotestosterone (DHT) ) Androgens are key stimulating factors for sebaceous gland development and sebum secretion.( Mainly DHT ) Binding androgen receptor ( AR ) This leads to conformational changes in the AR (alcoholic acid) receptor, which directly induces lipid synthesis enzymes in the sebaceous glands. ( Such as FASN, ACC ) The expression of [a substance] indirectly activates the PI3K / AKT-mTORC1 axis and enhances SREBP1 activity.

[0006] Insulin / IGF-1 pathway: Insulin and IGF-1 are potent lipid-promoting signaling agents, primarily exerting their effects by activating the PI3K-AKT signaling cascade. Activated PI3K-AKT signaling can inhibit GSK3β. ( Stabilize SREBP1 protein ) Inhibit FOXO1 ( Relieve SREBP1 transcriptional repression ) And activate mTORC1 to amplify the SREBP1 function.

[0007] PPARγ pathway: PPARγ is a member of the nuclear receptor superfamily, and it particularly promotes the activity of wax esters. ( WEs ) and triglycerides ( TAGs ) Synthesis; upon activation, it forms a heterodimer with RXR, binding to the PPAR reaction element. ( PPRE ) Inducing participation in fatty acid uptake ( such as FATP ) Fatty acid activation ( ACS ) Triglyceride synthesis ( such as DGAT ) and key enzymes for wax ester synthesis ( For example, Awat2 ) PPARγ can induce the expression of SCD1 and has cross-talk and synergistic effects with SREBP1 in regulating lipid synthesis genes.

[0008] AMPK pathway: AMPK is a cellular energy sensor that is activated when energy is scarce, inhibiting energy-consuming processes. ( Such as lipid synthesis ) Promote the production capacity process ( Such as fatty acid oxidation ) Phosphorylation and inhibition of SREBP1c processing, maturation, and transcriptional activity, blocking SREBP1 cleavage activation, and comprehensively inhibiting de novo lipid synthesis.

[0009] SREBP1 acts as a regulator of de novo synthesis of fatty acids and triglycerides. (DNL ) The core transcription factor is first synthesized on the endoplasmic reticulum (ER). The precursor is transported to the Golgi apparatus by SREBP cleavage of the activating protein (SCAP). Then, it is cleaved by two proteases (Site-1 protease (S1P) and Site-2 protease (S2P)) to release its N-terminal active domain and enter the nucleus to exert its transcription factor function.

[0010] In summary, sebum synthesis has a complex mechanism involving multiple pathways and genes, and finding specific regulatory molecules is technically challenging. There is an urgent need in this field to find novel active molecules to achieve source regulation of sebum synthesis and effectively solve seborrhea. Summary of the Invention

[0011] The purpose of this invention is to provide a composition for reducing scalp oil, its preparation method, and its application.

[0012] In a first aspect of the invention, there is provided an application of a composition for preparing an agent that reduces scalp oiliness; said composition comprising effective amounts of betulin and glycyrrhizic acid; wherein betulin: 0.2 to 10 parts by weight; glycyrrhizic acid: 10 to 60 parts by weight.

[0013] In one or more embodiments, the composition contains 0.2 to 8 parts by weight of betulinol, preferably 0.5 to 6 parts by weight of betulinol.

[0014] In one or more embodiments, the composition contains 10-50 parts by weight of glycyrrhizic acid, preferably 10-40 parts by weight of glycyrrhizic acid.

[0015] In one or more embodiments, for example, the composition contains 0.6, 0.8, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 7, 8 or 9 parts by weight of betulinol.

[0016] In one or more embodiments, for example, the composition contains 12, 15, 20, 25, 30, 35, 40, 45, 50 or 55 parts by weight of glycyrrhizic acid.

[0017] In one or more embodiments, the composition contains effective amounts of betulinol and glycyrrhizic acid as active components, or effective amounts of betulinol and glycyrrhizic acid as the main active components (contributing the major contribution).

[0018] In one or more embodiments, the composition comprises betulin and glycyrrhizic acid.

[0019] In another aspect of the invention, a composition for reducing scalp oiliness is provided, comprising effective amounts of betulin and glycyrrhizic acid; wherein betulin: 0.2 to 10 parts by weight; glycyrrhizic acid: 10 to 60 parts by weight.

[0020] In one or more embodiments, the components betulin and glycyrrhizic acid in the composition are present in a mixed system (e.g., a formulation) in a diluted (e.g., diluted 1 to 50 times, specifically 5, 10, 20, 25, 30, 40 times) or concentrated (e.g., concentrated 1 to 50 times, specifically 5, 10, 20, 25, 30, 40 times) form (forming a mixed system).

[0021] In one or more embodiments, the composition includes 2, 3, 4, 5, 6, 7, 8 or 9 parts by weight of betulin.

[0022] In one or more embodiments, the composition comprises 11, 12, 13, 15, 18, 20, 25, 30, 35, 40, or 45 parts by weight of glycyrrhizic acid.

[0023] In one or more embodiments, the betulin:glycyrrhizic acid ratio is 1:1 to 16 by weight percentage; preferably 1:1.2 to 14; preferably 1:1.5 to 12; preferably 1:5 to 10 (e.g., 1:5, 1:6 or 1:8).

[0024] In another aspect of the invention, a method for preparing a composition for reducing scalp oiliness is provided, comprising: mixing effective amounts of betulinol and glycyrrhizic acid; wherein betulinol: 0.2 to 10 parts by weight; glycyrrhizic acid: 10 to 60 parts by weight.

[0025] In one or more embodiments, the composition contains 0.2 to 8 parts by weight of betulinol, preferably 0.5 to 6 parts by weight of betulinol.

[0026] In one or more embodiments, the composition inhibits the expression of sterol regulatory element-binding protein 1 (SREBP1), inhibits sebum secretion from sebaceous gland cells, and inhibits scalp sebum secretion.

[0027] In another aspect of the invention, a scalp protection formulation is provided, comprising the aforementioned composition for reducing scalp oiliness, and a pharmaceutically acceptable hair care adjuvant.

[0028] In one or more embodiments, the scalp protection preparation includes: shampoo preparations (such as shampoo, hair cream, etc.) and scalp protection preparations (including hair serum, scalp serum, hair conditioner, etc.).

[0029] In one or more embodiments, the hair adjuvants include adjuvants for preparing shampoo formulations and adjuvants for preparing scalp protection formulations.

[0030] In one or more embodiments, the scalp protection preparation can be formulated as an ointment, cream, lotion, water, gel, powder, etc.

[0031] In another aspect of the invention, a method for reducing scalp oiliness is provided, comprising: administering the composition or the scalp protection preparation to a desired subject.

[0032] In one or more embodiments, the method for reducing scalp oil is a non-therapeutic method.

[0033] In one or more embodiments, the object includes a person.

[0034] In one or more embodiments, the objects include non-human mammals, preferably including (but not limited to): non-human primates (such as monkeys, orangutans, etc.), rodents (including rats, mice, hamsters, etc.).

[0035] In another aspect of the invention, a package (box) (or kit) for reducing scalp oiliness is provided, comprising any of the compositions described above.

[0036] In another aspect of the invention, a package (box) (or kit) for reducing scalp oiliness is provided, comprising the aforementioned scalp protection preparation.

[0037] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Attached Figure Description

[0038] Figure 1 In Example 1 of this invention, the results of the cytotoxicity test of the test sample are shown; a is betulin, and b is glycyrrhizic acid.

[0039] Figure 2 In Example 2 of this invention, the inhibitory effects of each group of substances on the SREBP1 target are shown. The top row, from left to right, consists of: NC group, 0.004% betulinol, and 0.008% betulinol; the bottom row, from left to right, consists of: 0.016% glycyrrhizic acid, 0.004% betulinol + 0.016% glycyrrhizic acid, 0.008% betulinol + 0.016% glycyrrhizic acid, and PC group.

[0040] Figure 3 In Example 2 of this invention, the quantitative analysis chart of the inhibitory effect of each group of substances on the SREBP1 target was compared with the significance analysis of NC and PC.

[0041] Figure 4 In Example 3 of this invention, the lipid secretion inhibition effects of each group of substances are shown. From left to right, they are: NC group, 0.004% betulinol + 0.016% glycyrrhizic acid group, 0.008% betulinol + 0.016% glycyrrhizic acid group, and PC group.

[0042] Figure 5 In Example 3 of this invention, the effects of each group of substances on inhibiting sebum secretion were analyzed, and the significance of these effects was compared with those of NC and PC.

[0043] Figure 6 In Example 4 of this invention, the scalp oil content test results of the shampoo containing the composition provided by this invention are presented.

[0044] Figure 7 In Example 5 of this invention, a Meibometer graph showing the change in scalp oil content of the serum containing the composition provided by this invention is shown. Detailed Implementation

[0045] Through in-depth research and screening, the inventors have revealed a functional composition comprising effective amounts of betulin and glycyrrhizic acid, which significantly reduce scalp oiliness through synergistic and enhanced effects. This invention provides a novel and highly efficient solution for scalp oil control.

[0046] the term

[0047] As used herein, the term "composition (formula) of the present invention" refers to a substance containing effective amounts of betulin and glycyrrhizic acid as major active ingredients (components) as described herein. The weight of the active ingredients may be 0.00001-20% of the total weight of the composition, preferably 0.0001-10%, more preferably 0.005-5%. Dilution or concentration may be performed if necessary.

[0048] As used herein, the terms “containing” or “including” include “comprising”, “substantially consisting of”, and “consisting of”.

[0049] As used herein, the term "consistently made of / component of" means that the composition may contain, in addition to the essential ingredients or components (betulin and glycyrrhizic acid), minor components and / or impurities that do not affect the active ingredient. For example, it may contain additives commonly used in the art or other similar art.

[0050] As used herein, the term “pharmaceuticalally acceptable” refers to a substance that is suitable for human and / or animal use without excessive adverse side effects (such as toxicity, irritation, and allergic reactions), i.e., a substance with a reasonable benefit / risk ratio.

[0051] As used herein, the term "effective amount" refers to an amount that is functional or active in humans and / or animals and is acceptable to humans and / or animals.

[0052] As used herein, the term "pharmaceuticalally acceptable hair adjuvant (carrier)" refers to a carrier used to deliver the active component (effective component) of the compositions of the present invention into the body, including various excipients and diluents. This term refers to adjuvants (carriers) that are not essential active ingredients themselves, do not exhibit excessive toxicity upon application, and are suitable for application on hair and scalp. Suitable adjuvants (carriers) are well known to those skilled in the art.

[0053] As used herein, the term "unit formulation" refers to a formulation of the composition of the present invention prepared for a single dose for ease of application, including but not limited to various gels, ointments, powders, or emulsions. The unit formulation contains an amount of the composition of the present invention suitable for single-dose, daily, or time-based application.

[0054] As used in this article, "isomer" includes: geometric isomers, enantiomers, and diastereomers (such as cis-trans isomers and conformational isomers).

[0055] As used herein, "solvent" refers to a compound that carries solvent molecules; for example, the solvate may be a hydrate.

[0056] As used herein, "parts by weight" or "number of parts by weight" are used interchangeably, and the parts by weight can be any fixed weight expressed in milligrams, grams, or kilograms (e.g., 1 mg, 1 g, 2 g, 5 g, or 1 kg). For example, a composition consisting of 1 part by weight of component a and 9 parts by weight of component b can be a composition consisting of 1 gram of component a + 9 grams of component b, or 10 grams of component a + 90 grams of component b, etc. In the composition, the percentage content of a certain component = (number of parts by weight of that component / sum of the number of parts by weight of all components) × 100%. Therefore, in a composition consisting of 1 part by weight of component a and 9 parts by weight of component b, the content of component a is 10%, and the content of component b is 90%.

[0057] compound components

[0058] This invention discloses for the first time a composition containing effective amounts of betulin and glycyrrhizic acid that has an inhibitory effect on sterol regulatory element binding protein 1 (SREBP1). The combination of the two has a synergistic and enhancing effect, thereby effectively reducing scalp oil.

[0059] Betulin, also known as betulin, betulin, or betulin, is a triterpenoid compound (a lupin-type pentacyclic triterpenoid) extracted from birch bark. It has a relative molecular mass of 442.72, appears as white needle-like crystals, is insoluble in water, slightly soluble in moderately polar solvents such as ethyl acetate, ethanol, and methanol, and readily soluble in low-polarity solvents such as chloroform. It is widely found in birch bark, jujube kernels, pomegranate bark, and other substances, with a higher concentration found in dried birch bark.

[0060] The molecular formula of betulin is: C 30 H 50 O2, CAS No.: 473-98-3, structural formula as follows: .

[0061] Technicians have attempted to use birch bark extract on the skin, but as a complex mixture, its core oil-controlling active ingredient is unclear and its content is unstable. Furthermore, it contains a large amount of plant matrix unrelated to the oil-controlling target, resulting in low efficiency and uncontrollable effects in targeting and inhibiting sebum secretion. Therefore, developing a single active ingredient with a clearly defined target, strong efficacy, and stable oil-controlling effect is a necessary condition for this invention.

[0062] glycyrrhizic acid ( Glycyrrhizic Acid The molecular weight is 822.93 g / mol, and the CAS number is 1405-86-3. ) It is a pentacyclic triterpenoid saponin ( Composed of glycyrrhetinic acid and two molecules of glucuronic acid. ) Its structural formula is as follows:

[0063] As a classic natural soothing ingredient, glycyrrhizic acid's multi-target mechanism of action, including anti-inflammatory and cortisol-regulating effects, has been confirmed in published studies. In this research patent, computer-aided drug design (CADD) technology was used to simulate the interaction between the molecule and SREBP1. The results showed that glycyrrhizic acid and SREBP1 have two hydrogen bond interactions, with a distance of 3.54 Å and a bond energy of -0.5 kcal / mol, and 3.16 Å and a bond energy of -4.7 kcal / mol, respectively. The above molecular docking evidence well predicted the direct effect of glycyrrhizic acid on SREBP1, and further experiments verified that glycyrrhizic acid does indeed have a significant inhibitory effect on SREBP1.

[0064] Bioavailability is an important indicator for evaluating the quality and efficacy of cosmetic raw materials. Different solid forms have different molecular spatial arrangements, resulting in differences in solubility and dissolution rate, which directly affect the absorption and bioavailability of active ingredients in the body. Glycyrrhizic acid can effectively improve the solubility of betulin, thereby enhancing its bioavailability.

[0065] This invention utilizes the combination of betulinol and glycyrrhizic acid to achieve oil control through multiple pathways of SREBP1 transport and direct action. Furthermore, it enhances the bioavailability of the composition without altering the structure of the raw materials, thereby achieving a synergistic effect. In contrast, dipotassium glycyrrhizate, having already formed a salt, does not possess this effect.

[0066] As a preferred embodiment of the present invention, the betulin is extracted by solvent extraction from plants and further purified and filtered to obtain a solid powder with a content >95%.

[0067] As a preferred embodiment of the present invention, the glycyrrhizic acid can be prepared by methods known in the art, such as extraction from the roots and rhizomes of plants of the genus Glycyrrhiza (e.g., licorice, glycyrrhiza inflata, glycyrrhiza glabra) by solvent (e.g., water, dilute ammonia or dilute ethanol), concentration, acid precipitation (to obtain free acid), conversion to salt (e.g., ammonium salt, sodium salt) and crystallization purification.

[0068] Those skilled in the art will understand that, after learning the structure of the compounds of the present invention, the compounds of the present invention can be obtained by methods well known in the art or by methods disclosed in the embodiments of the present invention, using known raw materials, such as chemical synthesis or extraction from biological sources (e.g., animals or plants), all of which are included in the present invention.

[0069] In this invention, isomers, solvates, precursors, or pharmaceutically acceptable salts of the compound (betulin or glycyrrhizic acid) may also be used, provided they have the same or substantially the same function as the compound. The term "pharmaceutically acceptable salt" can refer to salts formed by reactions with inorganic acids, organic acids, alkali metals, or alkaline earth metals. These salts include (but are not limited to): (1) salts formed with inorganic acids such as hydrochloric acid, sulfuric acid, carbonic acid, nitric acid, and phosphoric acid; and (2) salts formed with organic acids such as hydrobromic acid, citric acid, tartaric acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, or p-toluenesulfonic acid. Other salts include salts formed with alkali metals or alkaline earth metals (such as sodium, potassium, calcium, or magnesium) in the form of esters, carbamates, or other conventional "prodrugs."

[0070] The compound has one or more asymmetric centers. Therefore, these compounds can exist as racemic mixtures, individual enantiomers, individual diastereomers, mixtures of diastereomers, or cis or trans isomers.

[0071] The term "precursor of the compound" refers to the precursor of the compound that, when taken by an appropriate method, is metabolized or chemically reacted in the patient's body to become betulin or glycyrrhizic acid, or a salt or solution of betulin or glycyrrhizic acid.

[0072] Composition

[0073] This invention provides a composition with oil-controlling effects, comprising: effective amounts of betulin and glycyrrhizic acid as the main active ingredients. This invention uses a combination of glycyrrhizic acid and betulin, utilizing the fact that glycyrrhizic acid promotes the dissolution of betulin to further enhance their synergistic effect, thereby effectively reducing the production of lipids in the skin.

[0074] In this invention, the dosage / ratio of betulin and glycyrrhizic acid was also optimized to obtain a suitable ratio that can very effectively inhibit the production of scalp oil without causing significant side effects.

[0075] In the preferred embodiment, betulin: 1-10 parts by weight; glycyrrhizic acid: 10-50 parts by weight.

[0076] The composition of the present invention with oil-controlling effect inhibits the synthesis of lipids in sebaceous gland cells through the synergistic effect of the two main components, thereby enhancing the oil-controlling effect of the composition in cosmetic applications.

[0077] It should be understood that when used to develop and prepare compositions, the formulation of the compositions may vary depending on the actual application. For example, they may be prepared in a concentrated or diluted form, and these variations should also be included in this invention.

[0078] The present invention also includes equivalents of the active ingredients listed in the table above, and formulations.

[0079] The compositions described in this invention or preparations containing the compositions can be in various forms; the possible forms include solids or liquids, as well as other forms such as semi-solids.

[0080] In some preferred embodiments, the composition may be a unit formulation. When the composition is prepared as a unit formulation, for example, one dose of the unit formulation may be applied daily or every other day; or one to two doses may be applied daily, depending on the amount of active ingredient in the unit formulation.

[0081] The preparation method of the composition of the present invention is determined according to the type of formulation to be prepared and the route of application.

[0082] Scalp protectant

[0083] This invention combines glycyrrhizic acid with betulin to obtain a composition with oil-controlling effects, which is a natural, safe, efficient and multifunctional scalp care solution, and applies it to the field of scalp protection.

[0084] Based on the novel findings of this invention, a scalp protection formulation is also provided, comprising a personal care product. The scalp protection formulation comprises the composition provided by this invention as described above and pharmaceutically acceptable hair care adjuvants.

[0085] In a preferred embodiment of the present invention, a composition is formed using glycyrrhizic acid and betulinol, the composition comprising 10-50 parts by weight of glycyrrhizic acid and 1-10 parts by weight of betulinol. After adding the composition to the oil phase and surfactant, the mixture is sealed, vortexed, and then shaken in a constant-temperature shaker at 30-50°C and 80-120 r / min for 36-54 h. After removal, the mixture is centrifuged at 4000-6000 r / min for 4-6 min, and the supernatant is collected as the drug-loaded oil phase and surfactant. The drug-loaded surfactant and co-surfactant are precisely measured and mixed to form a mixed surfactant. The mixture is magnetically stirred at 30-60°C and 1000-1100 r / min, and distilled water is added dropwise until the solution changes from turbid to transparent. Alternatively, the mixture can be combined using other carrier methods. Furthermore, the oil phase is a polar oil, and in this embodiment, isopropyl myristate is used, with a weight of approximately 10-30 parts. Furthermore, the surfactant is a nonionic surfactant, and in this embodiment, Tween 60 is used, with a weight of approximately 15-25 parts. Furthermore, the co-surfactant is a polyol, and in this embodiment, glycerol is used, with a weight of approximately 30-50 parts. Furthermore, the addition of distilled water ranges from approximately 20 to 30 parts; Furthermore, the composition provided by this invention can be mixed with a cosmetically acceptable carrier to obtain various cosmetics or personal care products that can be applied to human skin, including but not limited to shampoos and conditioners, skin care and makeup lotions, serums, creams, and lotions.

[0086] More specifically, for example, in Examples 4 and 5 of the present invention, two specific sets of hair care adjuvants are listed, one of which is mixed with the composition of the present invention to form a shampoo, and the other of which is mixed with the composition of the present invention to form a scalp serum.

[0087] Furthermore, the compositions of the present invention can be added to conventional shampoos, conditioners, hair serums, and other products to reduce scalp oiliness.

[0088] Compared with the prior art, the present invention has the following beneficial effects: This invention achieves a synergistic effect by combining glycyrrhizic acid and betulin, thereby enhancing the composition's lipid inhibition effect on sebaceous gland cells.

[0089] The invention will be better understood from the following examples. However, those skilled in the art will understand that the specific methods and results are merely for illustrating the invention and not for limiting it. Experimental methods in the following examples, unless otherwise specified, are generally performed under conventional conditions or as recommended by the manufacturer.

[0090] Unless otherwise stated, percentages in the examples are weight percentages (w / w %).

[0091] Test Example 1: Effective Concentration Test

[0092] (1) Cell seeding and culture

[0093] Human sebaceous gland cells (SZ95) were cultured at 5 × 10⁶ cells per well. 3 Cells were seeded at a density of 1,000 cells per well in 96-well culture plates and incubated at 37°C and 5% C. Incubate in a constant temperature incubator for 12 hours until the cells are completely adhered to the wall.

[0094] (2) Drug intervention

[0095] The original culture medium was discarded. For the betulin test group, sample solutions of varying concentrations (0.000089%, 0.00022%, 0.00044%, 0.00089%, 0.0022%, 0.0044%, 0.0089%, 0.022%, w / v) were added. For the glycyrrhizic acid test group, sample solutions of varying concentrations (0.00016%, 0.00041%, 0.00082%, 0.0016%, 0.0041%, 0.0082%, 0.016%, 0.041%, w / v) were added. The blank control group was supplemented with an equal volume of complete culture medium, and the cultures were continued for 24 hours.

[0096] (3) Cell viability assay

[0097] Add 10 μL of CCK-8 reagent to each well, incubate in the dark for 2 hours, and then measure the absorbance at 450 nm using a microplate reader to assess the biocompatibility of the sample and screen for cell-safe concentrations of the substance. A cell viability rate greater than 90% is considered a suitable concentration.

[0098] according to Figure 1 The cell viability of betulin was >92% at a concentration <0.0089%, while the cell viability was only 63.46% at a concentration of 0.022%; therefore, its concentration was determined to be no more than 0.0089%.

[0099] according to Figure 1 The survival rate of glycyrrhizic acid is >96% in the 0.016% concentration range, but drops significantly to 46.90% at a concentration of 0.041%; therefore, it is determined that a concentration below 0.016% is preferred.

[0100] Test Example 2: In vitro verification of the inhibitory effect of the composition on SREBP1 protein expression.

[0101] Materials: Human sebaceous gland cells ( SZ95 ) SREBP1 specific antibody.

[0102] Instruments: Optical microscope, confocal microscope.

[0103] (1) Trial grouping and intervention

[0104] Based on test example 1 ( Sample safety concentration test ) As a result, betulin sample A was selected. ( 0.004%, 0.008% ) Glycyrrhizic acid sample B ( 0.016% ) A ( 0.004% ) +B ( 0.016% ) A ( 0.008% ) +B ( 0.016% ) As an experimental group, a positive control group was also set up. ( PC, 1μM Fatostatin, SREBP1 pathway inhibitor ) and negative control group ( NC, culture medium only ) Each group consisted of three independent biological replicates. The culture medium used was SZ95 dedicated medium (product number bio-121741, Eosurg Biotechnology). The drug was added to the cell culture medium at the final concentration described above.

[0105] (2) Immunofluorescence staining

[0106] Cell fixation and permeabilization: 24 hours after intervention, the culture medium was discarded, the cells were gently washed 3 times with PBS, fixed with 4% paraformaldehyde at room temperature for 15 minutes, and permeabilized with 0.5% Triton X-100 for 10 minutes.

[0107] Antibody incubation: Add 5% BSA to block nonspecific sites for 30 minutes, then incubate with SREBP1 primary antibody sequentially.( 1:200, 4°C overnight ) and fluorescently labeled secondary antibodies ( 1:500, in a dark, room-temperature environment for 1 hour ) After each antibody incubation, wash three times with TBST for five minutes each time.

[0108] Fluorescence images were acquired using a fluorescence microscope, and the fluorescence intensity of SREBP1 was quantified using ImageJ software.

[0109] Figure 2 and Figure 3 Immunofluorescence quantitative analysis showed that the negative control group ( NC, mean = 88.37 ) There were highly significant differences compared to all treatment groups. ( p<0.0001 ) This indicates that the SREBP1 fluorescence intensity in the NC group was significantly higher than that in other experimental groups and the positive control group. ( PC ) Compared with the negative control group, the fluorescence intensity of each treatment group was significantly reduced: 0.004% A group decreased by 48.13%; 0.008% A group decreased by 64.54%; 0.016% B group decreased by 24.92%; 0.004% A+ 0.016% B group decreased by 72.42%; 0.008% A+ 0.016% B group decreased by 90.06%; and PC group decreased by 89.54%.

[0110] Table 1 shows the inhibition data of each group of substances against the SREBP1 target.

[0111] Table 1. Inhibition data of each group of substances against the SREBP1 target

[0112] Based on the above, the combination of betulin and glycyrrhizic acid showed a significantly higher inhibition rate compared to the single-use group, indicating that the combined use of betulin and glycyrrhizic acid formed a very significant synergistic effect.

[0113] Test Example 3: Sebum Secretion Detection

[0114] (1) Samples of 0.004%A + 0.016%B and 0.008%A + 0.016%B were selected as experimental groups, and a positive control group, Finasteride, was set up at the same time. ( PC, 10μM ) and blank control group ( NC, culture medium only ) Each group had 3 independent biological replicates. Human sebaceous gland cells ( SZ95 )Cells were cultured in SZ95 special medium (product number bio-121741, Eosur Bio) and treated with drugs.

[0115] (2) Nile Red Staining and Quantification

[0116] Lipid staining: 24 hours after drug intervention, the culture medium was removed, cells were washed twice with PBS, and fixed with 4% paraformaldehyde for 15 minutes. Under light-protected conditions, 10 μg / mL Nile Red staining solution was added to each well, and the cells were incubated at 37°C for 30 minutes in the dark.

[0117] Fluorescence detection: Free dye was removed by PBS rinsing, lipid droplet images were captured under a fluorescence microscope, and the intensity of red fluorescence was analyzed using ImageJ software.

[0118] according to Figure 4 and Figure 5 The effect of inhibiting sebum secretion showed that there were extremely significant differences between the negative control group and all treatment groups (p<0.0001).

[0119] The results showed that the fluorescence intensity in the NC group was significantly higher than that in other experimental groups and the positive control group (PC). The negative control was used as a baseline. ( The mean is 92.1 ) Among them, low combination ratio ( 0.004%A + 0.016%B ) The inhibition rate of sebum secretion was 58.31%, with higher combination ratios... ( 0.008%A + 0.016%B ) The inhibition rate of sebum secretion was 87.92%, compared to the positive control. ( PC, Fatostatin ) The inhibition rate was 87.62%. The inhibition rate of the combination was significantly higher than that of the single-use groups.

[0120] Table 2. Data on the inhibition of lipid secretion in each group

[0121] Test Example 4: Shampoo containing the composition of the present invention and its scalp oil content test

[0122] Test samples: Shampoo products containing the compositions of the present invention (the compositions of the present invention were added to the blank products in Table 3), and blank products without the compositions of the present invention (Table 3).

[0123] Table 3

[0124] Blank product preparation: Aqueous phase preparation ( 70-75℃) First, premix carbomer, and simultaneously dissolve disodium EDTA, sodium chloride, acrylamide propyltrimethylammonium chloride / acrylamide copolymer, add sodium lauryl ether sulfate, sodium cocoyl methyl taurate, and cocamidopropyl betaine and stir until transparent.

[0125] Heating premixed oil phase ( Optional ) : Cocoyl oleate monoethanolamide, cetearyl alcohol, ethylene glycol stearate.

[0126] Emulsification stage ( 50-60℃ ) Slowly add the oil phase to the water phase, homogenize and emulsify, and then add PEG-120 methyl glucoside ester to thicken.

[0127] cooling phase ( ≤45℃ ) Add guar gum hydroxypropyltrimethylammonium chloride, allantoin, polydimethylsiloxane alcohol (and) triethanolamine dodecylbenzenesulfonate, PEG-90M, sodium benzoate, phenoxyethanol, and ethylhexylglycerin.

[0128] Final adjustment ( room temperature ) Adjust the pH to slightly acidic using citric acid / sodium hydroxide. ( 5.5-6.0 ) ,join in ( Daily use ) The flavoring is adjusted for viscosity with a small amount of sodium chloride.

[0129] Experimental product preparation: During the cooling stage, the combination of betulinol and glycyrrhizic acid of the present invention was added to achieve a final concentration of betulinol 0.01% and glycyrrhizic acid 0.04%.

[0130] Test instrument: Sebumeter ( SM815, Courage+Khazaka, Germany ) .

[0131] Test time points: before product use, immediately after a single product use, 24 hours after a single product use, 48 hours after a single product use, and [other times].

[0132] 72 hours after first use, 2 weeks after product use, and 4 weeks after product use.

[0133] Data processing: Statistical analysis was performed using STATISTIC 7.1 software with a significance level of α=0.05 and a two-tailed test. The mean, median, standard deviation, maximum and minimum values ​​of each test parameter were calculated. The t-test method was applied to analyze the changes in the product before and after use.

[0134] The results of the Sebumeter scalp oil change value test are shown in Table 4. The scalp oil content test results of the shampoo containing the composition provided by this invention are as follows: Figure 6 .

[0135] Table 4. Results of Sebumeter scalp oil change test (%)

[0136] The results showed that compared with before product use, the scalp oil content in the test area was significantly reduced immediately after a single use, 24 hours after a single use, 48 hours after a single use, 72 hours after a single use, 2 weeks after product use, and 4 weeks after product use. The percentage reductions were 86.23%, 52.26%, 35.19%, 10.98%, 28.93%, and 36.18%, respectively. The scalp oil content was 0.14 times, 0.48 times, 0.65 times, 0.89 times, 0.71 times, and 0.64 times that before product use, respectively.

[0137] Compared with before product use, the percentage change in scalp oil content for the individual subjects with the greatest change was 95.97%, 70.18%, 55.97%, 34.279%, 55.54%, and 74.03% immediately after a single product use, 24 hours after a single product use, 48 hours after a single product use, 72 hours after a single product use, 2 weeks after product use, and 4 weeks after product use, respectively. The scalp oil content of the individual subjects with the greatest change was 0.04 times, 0.30 times, 0.44 times, 0.65 times, 0.44 times, and 0.26 times that before use, respectively.

[0138] Test Example 5: Scalp serum containing the composition of the present invention and its scalp oil content test

[0139] Test samples: scalp serum products containing the compositions provided by the present invention (the compositions of the present invention are added to the blank products in Table 4), and blank products without the compositions of the present invention (Table 5).

[0140] Table 5

[0141] Blank product preparation: Phase A: Water, Glycerin, Butylene Glycol, 1,2-Pentanediol, Sodium Hyaluronate, 1,3-Propane Diol, Acrylic Acid ( ester ) Class / C10-30 Alkyl Acrylate Crosslinked Polymer.

[0142] Phase B: Polydimethylsiloxane, glycerol triglyceride ( Ethylhexanoic acid) Ester, pentaerythritol tetra ( Ethylhexanoic acid ) Ester, PEG-11 methyl ether polydimethylsiloxane, C13-14 isoparaffins, octanoic acid / decanoic acid triglycerides.

[0143] Phase C: Water, potassium hydroxide.

[0144] Phase D: Water, panthenol.

[0145] Phase E: Phenoxyethanol, Ethylhexylglycerol.

[0146] Heat phase B to 75-80℃ and disperse and stir until homogeneous, then keep warm. Heat phase A to 75-80℃ and disperse until homogeneous, then add phase B to phase A while stirring, and homogenize at 2000-4000 rpm for 5-10 minutes. Cool to 45℃, add phase C and stir for 5 minutes, then add phases D and E and stir until homogeneous. Add flavoring.

[0147] Experimental product preparation: During the cooling stage, the combination of betulin and glycyrrhizic acid of the present invention was added to make the final concentration of betulin 0.02% and glycyrrhizic acid 0.08%.

[0148] Test instrument: Meibometer MB560 (Courage+Khazaka, Germany).

[0149] Data processing: Statistical analysis was performed using STATISTIC 7.1 software with a significance level of α=0.05 and a two-tailed test. The mean, median, standard deviation, maximum and minimum values ​​of each test parameter were calculated. The t-test method was applied to analyze the changes in the product before and after use.

[0150] The results of the Meibometer scalp oil change test are shown in Table 6. The graph showing the change in Meibometer scalp oil values ​​of the serum containing the composition provided by this invention is shown below. Figure 7 .

[0151] Table 6. Meibometer test results of scalp oil change (transmittance, in relative value)

[0152] 1. vs {D0 ( 1 hour after washing hair ) -D0 ( Immediately after washing hair ) Rate of change ( % ) = ( Average of test values ​​for the nth hour / day - {D0} ( 1 hour after washing hair) -D0 ( Immediately after washing hair )} test value mean) / {D0 ( 1 hour after washing hair ) -D0 ( Immediately after washing hair ) Test value mean 100% ("vs") This refers to the comparison with immediately after washing hair on the same day. ) .

[0153] 2. Interpretation of significance markers: "Unmarked" indicates no statistically significant difference; p<0.05 indicates a significant difference; where, "This means 0.01 ≤ p < 0.05;" " indicates that 0.001 ≤ p < 0.01.

[0154] The results showed that compared with before product use, and with D0 ( 1 hour after washing hair ) In comparison, the scalp sebum secretion rate of the product group was higher on day 12. ( 1 hour after washing hair ) D14 ( 1 hour after washing hair ) D28 ( 1 hour after washing hair ) Significantly reduced ( p<0.001 ) .

[0155] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the scope of protection of the present invention. Therefore, the scope of protection of this patent should be determined by the appended claims. Furthermore, all documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference.

Claims

1. The application of a composition, characterized in that, This formulation is used to prepare an agent for reducing scalp oiliness; the composition comprises effective amounts of betulin and glycyrrhizic acid; wherein betulin: 0.2 to 10 parts by weight; glycyrrhizic acid: 10 to 60 parts by weight; and the composition uses betulin and glycyrrhizic acid as active ingredients.

2. The application as described in claim 1, characterized in that, The composition contains 0.2 to 8 parts by weight of betulin.

3. The application as described in claim 1, characterized in that, The composition contains 10-50 parts by weight of glycyrrhizic acid.

4. A composition for reducing scalp oiliness, comprising effective amounts of betulin and glycyrrhizic acid; wherein, Birch resin Alcohol: 0.2 to 10 parts by weight; glycyrrhizic acid: 10 to 60 parts by weight; and the composition uses betulin and glycyrrhizic acid as active ingredients.

5. The composition for reducing scalp oiliness as described in claim 4, characterized in that, The components betulinol and glycyrrhizic acid in the composition exist in the mixed system in a diluted or concentrated form.

6. A method for preparing a composition for reducing scalp oiliness, comprising: The effective amounts of betulinol and glycyrrhizic acid are mixed; wherein, betulinol: 0.2 to 10 parts by weight; glycyrrhizic acid: 10 to 60 parts by weight; and the composition uses betulinol and glycyrrhizic acid as active ingredients.

7. The composition according to any one of claims 4-5 or the method according to claim 6, characterized in that, The composition contains 0.2 to 8 parts by weight of betulin.

8. The application as described in claim 1, the composition as described in any one of claims 4-5, or the method as described in claim 6, characterized in that, The composition inhibits the expression of sterol regulatory element-binding protein 1, inhibits sebum secretion from sebaceous gland cells, and inhibits scalp sebum secretion.

9. A scalp protection preparation comprising: The composition for reducing scalp oil according to any one of claims 4 to 5; And pharmaceutically acceptable excipients.

10. A method for reducing scalp oiliness, comprising: The composition of any one of claims 4 to 5 or the scalp protection preparation of claim 9 is given to the desired subject.

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