TW201924673A - Cholesterol synthesis promoter - Google Patents

Abstract

Provided is a new cholesterol synthesis promoter. This cholesterol synthesis promoter uses, as an effective component, at least one component selected from the group consisting of Uncaria gambir extracts, copolymers having a 2-methacryloyloxyethyl phosphorylcholine group, retinol acetate, and Psophocarpus tetragonolobus extracts.

Description

Cholesterol synthesis accelerator

The present invention relates to a novel cholesterol synthesis promoter, a composition comprising the cholesterol synthesis promoter for promoting differentiation of epidermal cells, and a method for screening for an agent for promoting differentiation of epidermal cells.

Aging is carried out in the organs of the whole body. For the skin that is visible to the naked eye, especially the face that is easy to concentrate, the wrinkles and fine lines that occur at the same time as aging plague most of the elderly, especially women. Recently, not only women, but also the desire of men to look young has become strong, and it is particularly desirable to alleviate wrinkles in the skin that accompany aging.

In response to this requirement, there has been an injection of botulinum toxin or hyaluronic acid, but in this treatment, it is respectful in terms of the injection of foreign matter, and also in terms of the sustainability of the effect. dissatisfied. In recent years, in place of such treatments, a treatment called "Platelet Rich Plasma (PRP) therapy" in which platelet components in the blood itself are concentrated and injected into the affected part of the body is being promoted in the aesthetic medical field. PRP therapy has been the research or clinical application technology since around 1985. It was mainly used in bone regeneration induction therapy for dental implants, but it is gradually applied to cosmetic medicine, mainly for wrinkles or relaxation. Elimination. Since PRP therapy returns to its own blood, it is not far from being respectful in terms of foreign body injection, and it is known to be long-lasting in terms of effects. It is considered that PRP contains various factors derived from platelets, which cooperate with serum fibrin to activate fibroblasts in the dermis and promote collagen production, which is beneficial to moisturized and elastic skin. At present, PRP is not limited to the dermatological region, and it is also applicable to liver regeneration treatment or myocardial infarction treatment for patients who have extracted most of the liver due to liver cancer (Non-Patent Document 1, Non-Patent Document 2, Non-Patent Document 3). Further, PRP (W-PRP) containing a large amount of white blood cells has been reported to be more effective than the prior PRP technology (Non-Patent Document 1 and Non-Patent Document 2), and it is useful for improvement of skin aging (Non-Patent Literature) 4. Non-Patent Document 5, Non-Patent Document 6).

However, the therapeutic effect of W-PRP is quite different from individual, and the treatment mechanism of W-PRP is still not well understood.
[Previous Technical Literature]
[Non-patent literature]

[Non-Patent Document 1] Classification of platelet concentrates: from pure platelet-rich plasma (P-PRP) to leucocyte- and platelet-rich fibrin (L-PRF). Dohan Ehrenfest DM, Rasmusson L, Albrektsson T. Trends Biotechnol. 2009 Mar; 27 (3): 158 - 67. Epub 2009 Jan 31. Review. PMID: 19187989
[Non-Patent Document 2] The basis of autologous W-PRP injection therapy for the purpose of wrinkle removal to the clinic Chuan Tiangang, Jin Xueqa, Journal of the National Society of Clinical Dermatology, 17, 1-26, 2008
[Non-Patent Document 3] Treatment of Depression: About Multiplatelet Injection, et al., MB Derma, 168: 29 - 35, 2010
[Non-Patent Document 4] Study on the effect of skin aging improvement of white blood cell-containing platelet-rich plasma (W-PRP), et al., 28th Cosmetic Skin Science Society, August 2010
[Non-Patent Document 5] Study on the Mechanism of Skin Aging Improvement of White Blood Cell-containing Platelet Plasma (W-PRP), et al., The 29th Cosmetic Dermatology Society, September 2011
[Non-Patent Document 6] Study on the mechanism of skin aging improvement including white blood cell platelet-rich plasma (W-PRP), The 31st Cosmetic Dermatology Society, August 2013
[Non-Patent Document 7] Free Radic Biol Med. 2006 Jul 15; 41 (2): 339 - 46

[The problem that the invention wants to solve]

An object of the present invention is to provide a novel cholesterol synthesis accelerator, a composition comprising the cholesterol synthesis promoter for promoting differentiation of epidermal cells, and a method for screening for a drug for promoting differentiation of epidermal cells.
[Technical means to solve the problem]

The inventors of the present invention investigated the distribution of gene expression in the skin before and after treatment of W-PRP using a microarray, and found that the expression of the DHCR7 gene was significantly increased by W-PRP treatment. Based on this finding, the inventors of the present invention conducted intensive studies, and as a result, it was surprisingly found that the expression of the DHCR7 gene in skin cells is closely related to the differentiation of epidermal cells, and further, the catechin extract has 2-methylpropene. The copolymer of methoxyethylphosphocholine, the retinol acetate, and the Winged Bean Extract promote the expression of the DHCR7 gene, thereby completing the present invention.

Therefore, this case contains the following inventions:
[1] A cholesterol synthesis promoter comprising a copolymer selected from the group consisting of catechin extract, 2-methylpropenyloxyethylphosphocholine, retinyl acetate, and Winged Bean Extract At least one component of the group consisting of.
[2] A composition for promoting differentiation of epidermal cells, comprising the cholesterol synthesis promoter according to 1.
[3] A method for screening for a drug for promoting differentiation of epidermal cells, wherein when a candidate agent is applied to a cultured cell, a drug which promotes the expression of the DHCR7 gene of the cell is selected as a drug for promoting differentiation of epidermal cells.
[4] The method according to 3, wherein the cultured cells are human epidermal keratinocytes.
[5] In the preparation of a composition for promoting differentiation of epidermal cells, selected from the group consisting of catechin extract, a copolymer having 2-methylpropenyloxyethylphosphocholine, and retinyl acetate And use of at least one component of the group consisting of the Winged Bean Extract.
[6] A composition for promoting differentiation of epidermal cells, which comprises an agent which promotes the expression of the DHCR7 gene.
[7] A cosmetic or therapeutic method for promoting differentiation of epidermal cells, which comprises applying a medicament for promoting the expression of the DHCR7 gene to a subject.
[8] Use of a medicament for promoting the expression of the DHCR7 gene in the preparation of a composition for promoting differentiation of epidermal cells.
[Effects of the Invention]

According to the present invention, differentiation of epidermal cells can be promoted, thereby preventing, ameliorating or treating a state or disease caused by inhibition of differentiation of epidermal cells.

The DHCR7 gene is a gene encoding a 7-dehydrocholesterol reductase (EC 1.3.1.21), which is mainly known as an endoplasmic reticulum membrane in cells, which is a well-known enzyme that converts 7-dehydrocholesterol into cholesterol. The DHCR7 gene is well known as the causative gene of Smith-Lemli-Opitz Syndrome (SOLS). In addition to symptoms such as mental retardation, facial deformity, toe and toe, and forebrain non-cracking deformity (severe cases), patients with SOLS also reported hypersensitivity to UVA (Ultraviolet A), which is considered to be The reason is that the 7-dehydrocholesterol in the blood is increased, and the metabolite of UVA absorption is increased to generate active oxygen (Non-Patent Document 7).

In one aspect of the present invention, there is provided a cholesterol synthesis accelerator comprising a copolymer selected from the group consisting of catechin extract, 2-methylpropenyloxyethylphosphocholine, and retinyl acetate. And at least one component of the group consisting of the Winged Bean Extract.

The catechin extract is derived from an extract of catechin (scientific name: Uncarina gambir) which is a plant belonging to the genus Rubiaceae in Southeast Asia, and examples of the extracting material, for example, a flower (bud) portion, a leaf portion, Branches, bark parts, roots, etc., but preferably leaf parts. The catechu extract can be obtained by a conventional method. For example, the portion used as the extraction raw material is dried as needed, and then immersed in an extraction solvent for a fixed period of time, or may be contacted with an extraction solvent heated under reflux, followed by filtration and concentration. The extraction solvent can be used arbitrarily as long as it is a solvent used in usual extraction, and examples thereof include alcohols such as water, methanol, ethanol, propylene glycol, 1,3-butylene glycol, and glycerin, chloroform, dichloroethane, and tetra. The organic solvents such as carbon chloride, acetone, and ethyl acetate are used singly or in combination as appropriate. The extract obtained by extracting with the above solvent may be used as it is, or the extract obtained by concentration may be used, or an extract may be used to remove impurities by an adsorption method such as an ion exchange resin; or adsorbed on After the column of the porous polymer (for example, Amberlite XAD-2) is dissolved in methanol or ethanol and concentrated. Further, an extract obtained by a distribution method such as water/ethyl acetate extraction or the like can also be used. The extract obtained in this manner can be directly used in the present invention, or further diluted with ethanol or the like, or dried, and the dried product can be directly used in the present invention, or the dried product can be used, for example, in ethanol to be used in the present invention.

The copolymer having 2-methylpropenyloxyethylphosphocholine is a copolymer of 2-methylpropenyloxyethylphosphocholine and a hydrophobic monomer having a weight average of 600,000 to 700,000. Molecular weight. For example, since the copolymer of 2-methacryloxyethylphosphoethylcholine and butyl methacrylate (MPC copolymer) has excellent moisture retention, it is used for hand cream or contact lens preservation solution, shampoo Fine, moisturizing milk, etc.

The retinyl acetate is an acetate of vitamin A and is used as a cosmetic ingredient such as a moisturizing cream.

The extract of the Winged Bean is an extract of the Winged Bean (scientific name: Psophocarpus tetragonolobus) which is also grown in Okinawa Prefecture or Ogasawara Islands in Japan, and is used as a raw material for extracting, for example, a seed. , pods, leaves, flowers, roots, whole plants, etc., but preferably seeds or pods. The Winged Bean Extract can also be obtained by the conventional method described above.

As a further aspect of the present invention, there is provided a composition comprising the above cholesterol synthesis promoter for promoting differentiation of epidermal cells. Such a composition can be used as a cosmetic composition or a pharmaceutical composition, and is effective for preventing, ameliorating or treating a state or disease caused by abnormal differentiation of epidermal cells caused by a decrease in local cholesterol synthesis, such as wrinkles, relaxation, etc. The age of the skin is reduced by cholesterol.

The composition of the present invention can be appropriately determined in amount, usage, and dosage form depending on the purpose of use. For example, the form of administration of the composition of the present invention is not particularly limited, and may be oral, parenteral, external or the like, and is preferably a composition for external use on skin. Examples of the dosage form include an external preparation such as an ointment, a cream, an emulsion, a lotion, a dressing, and a bath; a parenteral preparation such as an injection, a drip, or a suppository; or a lozenge, a powder, or a capsule. Oral administration agents such as granules, extractants, syrups and the like.

The compounding amount of the active ingredient of the composition of the present invention can be appropriately determined depending on the use, but in the total amount of the composition, the catechu extract is typically 0.0001% to 10%, preferably 0.001% to 1%, The optimum is 0.05% to 0.5%. The copolymer having 2-methylpropenyloxyethylphosphocholine is typically formulated in an amount of 0.0001% to 10%, preferably 0.001% to 1%, most preferably 0.05% to 0.5%. The retinyl acetate is usually formulated in an amount of 0.000001% to 1%, preferably 0.00001% to 0.01%, most preferably 0.00005% to 0.005%. The Winged Bean Extract is typically formulated at 0.0001% to 10%, preferably 0.001% to 1%, most preferably 0.05% to 0.5%.

Further, in the composition of the present invention, in addition to the above-mentioned active ingredients, for example, a whitening agent, a moisturizing agent, an oily component, an ultraviolet absorber, a surfactant, a tackifier, or the like which are usually used in cosmetics or pharmaceuticals may be appropriately formulated. Alcohols, powder components, colorants, aqueous components, water, various skin nutrients, and the like.

Further, when the composition of the present invention is used as a skin external preparation, an auxiliary agent conventionally used in an external preparation for skin, such as disodium edetate, trisodium ethylenediaminetetraacetate, citric acid, may be appropriately formulated. Metal blockers such as sodium, sodium polyphosphate, sodium metaphosphate, gluconic acid; hot water extracts of caffeine, tannin, verapamil, tranexamic acid and its derivatives, licorice, papaya fruit, Various natural medicines, vitamin E acetate, glycyrrhizic acid and its derivatives or their salts; vitamin C, magnesium ascorbyl phosphate, ascorbyl glucoside, arbutin, kojic acid and other whitening agents; glucose, fructose, mannose, Sugars such as sucrose and trehalose.

In another aspect of the present invention, there is provided a method for screening for an agent for promoting differentiation of epidermal cells using the hyperactivity of the DHCR7 gene as an indicator. The hyperactivity of the DHCR7 gene can be determined, for example, by directly measuring the amount of DHCR7 in the cultured cells. The cultured cells are typically skin cells, especially epidermal cells, containing keratinocytes, granulosa cells, spine cells, etc., and may be from humans, such as rats, mice, rabbits, and the like. In the case of directly measuring the amount of DHCR7, it is preferred to use an antibody specific for DHCR7, which can be obtained by methods well known in the art, such as immunostaining using fluorescent substances, pigments, enzymes, etc., Western blotting method. The immunoassay method is carried out by various methods such as an ELISA method and an RIA method. Alternatively, it may be determined by extracting RNA from the cultured cells and measuring the amount of mRNA encoding DHCR7. The extraction of mRNA and the determination of its amount are also well known in the art. For example, the quantification of RNA is carried out by quantitative polymerase chain reaction (PCR), such as real-time polymerase chain reaction (RT-PCR). Selection of primers suitable for RT-PCR can be carried out by methods well known in the art.

In the RT-PCR analysis of the DHCR7 gene, for example, the following primers can be used.
Forward primer: 5'-AGGGGAAGGTGGGCGCAGGAC-3' (sequence number 1)
Reverse primer: 5'-TTGGGCCCTCCAGCCTCTTGC-3' (sequence number 2)

In the above measurement, for example, if the expression of the DHCR7 gene in the cultured cells is significantly different from the control value, it is preferably 30% or more, more preferably 50% or more, still more preferably 70% or more, and most If it is more than 100%, it can be judged as "a drug that promotes differentiation of epidermal cells." The control value is not limited, and may be, for example, an average value of the expression amount of the DHCR7 gene of the cultured cells in the corresponding portion of a statistically significant number of healthy persons.

Using the above screening method, a novel agent that promotes differentiation of epidermal cells can be easily obtained.

The present invention will be further specifically described by the following examples. Furthermore, the present invention is not limited to this.
[Examples]

1. Microarray analysis of genes expressing hyperactivity by W-PRP treatment. W-PRP treatment was performed on the anterior side of the upper arm of two subjects, and 5 mm was used before treatment, after 2 weeks of treatment, and after 3 months of treatment. The puncher performs the collection of skin tissue. Each sample was disrupted in a liquid nitrogen by a freeze-shear crushing device, and RNA in the cells was extracted with Isogen (Nippon Gene), and a labeled probe was synthesized according to a guide (Agilent) to spot about 40,000 genes. Hybridization was performed on a DNA microarray (one-color, whole mouse 44K, Agilent). The resulting signal was detected using a scanner Agilent dual-laser Microarray scanner G2565AA and quantified using the analytical software Feature Extraction Software 9.1 (Agilent) as a gene performance indicator value.
As shown in Fig. 1, it was found that the performance of DHCR7 was improved by W-PRP processing.

2. Effect of differentiation of epidermal keratinocytes by DHCR7 inhibitor Human epidermal keratinocytes were subcultured in CnT-07 medium (CELLnTEC), and cells of the fourth passage were suspended in the same medium in collagen. The cells were seeded in a 6-well multi-well plate (Asahi Glass) at 2 × 10 ⁵ cells/well, and cultured at 37 ° C for 3 to 5 days in the presence of 5% CO ₂ until the cells became dense. Replace with CnT-02 medium (CELLnTEC) to initiate differentiation induction. At the same time, a DHCR7 inhibitor (AY-9944) (Sigmα-Aldrich) dissolved in dimethyl hydrazine (DMSO) was added at a final concentration of 10 μM, and cultured for 4 days. The state observation of the cells was carried out under a phase contrast microscope. As a control, a sample in which an equal amount (final concentration of 0.1%) of DMSO was added was used.
As shown in the photograph of Fig. 2, morphological changes accompanying differentiation of epidermal cells were inhibited by inhibition of DHCR7.

3. Changes in expression of epidermal keratinocyte differentiation markers caused by DHCR7 inhibitors from cultured cells using RNA extraction reagent MagNA Pure LC mRNA HS kit (Roche) and automatic nucleic acid extraction device MagNA Pure LC 1.0 Instruments (Roche Company), according to the instructions provided, the extraction and purification of mRNA. With respect to the above DHCR7 inhibitor (AY-9944) and a control, cDNA was synthesized using a reverse hexamer/initiator from RNA using reverse transcriptase Superscript II (Invitrogen) according to the instructions of Invitrogen. Using the synthesized cDNA as a template, keratin 10 as a differentiation marker was carried out using the primer pair of SEQ ID NOs: 1 and 2 described later, the reaction reagent QuantiFast SYBR Green RT-PCR kit (Qiagen), and the reaction device LightCycler (Roche). Quantitative real-time PCR of genes. The composition conditions are in accordance with Qiagen's operating instructions. Further, the conditions of the PCR were initially modified at 95 ° C for 10 minutes, modified at 95 ° C for 10 seconds, annealed at 60 ° C for 10 seconds, and elongated at 72 ° C for 12 seconds. Furthermore, G3PDH was used as an internal standard to correct the amount of mRNA in the control group.
Keratin 10:
Forward introduction: 5'-CCATCGATGACCTTAAAAATCAG-3' (sequence number 1)
Reverse primer: 5'-GCAGAGCTACCTCATTCTCATACTT-3' (sequence number 2)
G3PDH:
Forward introduction: 5'-GCACCGTCAAGGCTGAGAAC-3' (sequence number 3)
Reverse primer: 5'-ATGGTGGTGAAGACGCCAGT-3' (sequence number 4)
As shown in the graph of Fig. 3, keratin 10, which is a differentiation marker for epidermal cells, was significantly reduced by AY-9944.

4. Determination of Cellular Cholesterol Content The solvent was evaporated by 200 μl of chloroform:isopropanol:NP-40 (7:11:0.1) after the cells on the first and fourth days after the start of differentiation described above. It was suspended in a cholesterol assay buffer contained in a cholesterol/cholesterol ester quantitative kit (BioVision) as a measurement sample. A sample was added to a 96-well plate attached to the kit, 50 μl of the reaction mixture was added to each well, and after incubating at 37 ° C for 1 hour, the absorbance at a wavelength of 570 nm was measured using a plate reader. A calibration curve was prepared using the kit's attached standard to quantify the cholesterol concentration in the sample.
As shown in Fig. 4A, the amount of cholesterol contained in the epidermal cells was reduced by AY-9944.

5. Measurement of cell respiration activity The cell culture solution on the first day after the start of differentiation prepared above was replaced with CnT-02 medium (CELLnTEC) containing 10% of alamarBlue (R) Cell Viability Reagent (invitrogen). After incubation at 37 ° C for 3 hours in the presence of 5% CO ₂ , the measurement was carried out using a fluorescence plate reader at an excitation wavelength/fluorescence wavelength: 540/590 nm.
As shown in Fig. 4B, no cell proliferation was observed due to AY-9944.
From the results shown in Figs. 4A and 4B, it was confirmed that the decrease in keratin 10 in Experiment 2 was not caused by cell death, and the cholesterol content of the cells was surely reduced by inhibiting DHCR7.

6. Cholesterol production enzyme (DHCR7) production promoted human epidermal keratinocytes were subcultured in CnT-07 medium (CELLnTEC), and the passaged passage 3 cells were suspended in the same medium in collagen-coated 12-well multi-well plates. (Asahi Glass) was sown at a ratio of 50,000, and cultured at 37 ° C for 3 to 5 days in the presence of 5% CO ₂ until the cells became dense. 30 kinds of candidate chemicals were used as the sample to be evaluated, and the CnT-07 medium in which each sample was added or the same amount of DMSO as a solvent for dissolving each of the evaluation target samples was added was added to the sample. Day culture. From the cultured cells, mRNA extraction and purification were performed using an RNA extraction reagent MagNA Pure LC mRNA HS kit (Roche) and an automatic nucleic acid extraction device MagNA Pure LC 1.0 Instruments (Roche) according to the instructions provided. For each sample, a cDNA was synthesized using a reverse transcriptase Superscript II (Invitrogen) from RNA using a random hexamer/introduction according to the Invitrogen protocol. The synthesized cDNA was used as a template, and the quantitative real-time PCR of the DHCR7 gene was carried out using the primer pair of SEQ ID NOs: 1 and 2 described later, the reaction reagent QuantiFast SYBR Green RT-PCR kit (Qiagen), and the reaction apparatus LightCycler (Roche). The composition conditions are in accordance with Qiagen's operating instructions. Further, the conditions of the PCR were initially modified at 95 ° C for 10 minutes, modified at 95 ° C for 10 seconds, annealed at 60 ° C for 10 seconds, and elongated at 72 ° C for 12 seconds. Furthermore, G3PDH was used as an internal standard to correct the amount of mRNA in the control group.
DHCR7:
Forward primer: 5'-AGGGGAAGGTGGGCGCAGGAC-3' (sequence number 1)
Reverse primer: 5'-TTGGGCCCTCCAGCCTCTTGC-3' (sequence number 2)
G3PDH:
Forward introduction: 5'-GCACCGTCAAGGCTGAGAAC-3' (sequence number 3)
Reverse primer: 5'-ATGGTGGTGAAGACGCCAGT-3' (sequence number 4)
As a result, it was found that catechin extract (Maruzen Pharmaceutical Co., Ltd.), MPC copolymer (Japanese fat (stock)), retinyl acetate (BASF), and Winged bean extract (one pill Pharcos) The four agents have a DHCR7 production promoting effect (Fig. 5).

7. Epidermal differentiation marker production caused by catechin extract promotes subculture of human epidermal keratinocytes in CnT-07 medium (CELLnTEC), and suspends passage 3 cells in the same medium for collagen coating. The cloth was sown at a ratio of 50,000 in a 12-well multi-well plate (Asahi Glass), and cultured at 37 ° C for 3 to 5 days in the presence of 5% CO ₂ until the cells became dense. The CnT-07 medium supplemented with catechin extract (Maruzen Pharmaceutical Co., Ltd.) or an equal amount of DMSO was added as a control, and the culture was further carried out for 1 day. From the cultured cells, mRNA extraction and purification were performed using an RNA extraction reagent MagNA Pure LC mRNA HS kit (Roche) and an automatic nucleic acid extraction device MagNA Pure LC 1.0 Instruments (Roche) according to the instructions provided. For each sample, a cDNA was synthesized using a reverse transcriptase Superscript II (Invitrogen) from RNA using a random hexamer/introduction according to the Invitrogen protocol. The synthesized cDNA was used in the template, and the primer pair of SEQ ID NO: 1 and 2 described later, the reagent QuantiFast SYBR Green RT-PCR kit (Qiagen), and the reaction device LightCycler (Roche) were used as the set of differentiation markers. Membrane protein (involucrin) Genetic W-PRP treatment Quantitative real-time PCR for microarray analyzers that perform hyperactive genes. The composition conditions are in accordance with Qiagen's operating instructions. Further, the conditions of the PCR were initially modified at 95 ° C for 10 minutes, modified at 95 ° C for 10 seconds, annealed at 60 ° C for 10 seconds, and elongated at 72 ° C for 12 seconds. Furthermore, G3PDH was used as an internal standard to correct the amount of mRNA in the control group.
Envelope protein:
Forward introduction: 5'-CTGCCTCAGCCTTACTGTGA-3' (sequence number 5)
Reverse primer: 5'-TGGGTATTGACTGGAGGAGG-3' (sequence number 6)
G3PDH:
Forward introduction: 5'-GCACCGTCAAGGCTGAGAAC-3' (sequence number 3)
Reverse primer: 5'-ATGGTGGTGAAGACGCCAGT-3' (sequence number 4)
As shown in Fig. 6, it was confirmed that the production of the envelope protein was promoted by the catechin extract.

Fig. 1 is a graph showing changes in the expression of the DHCR7 gene before, after 2 weeks of treatment, and after 3 months of treatment of W-PRP.

Fig. 2 is a photograph showing changes in morphology accompanying differentiation of human epidermal keratinocytes by a DHCR7 inhibitor (AY-9944).

Fig. 3 is a graph showing changes in the expression of the epidermal cell molecular marker (keratin 10) caused by the DHCR7 inhibitor (AY-9944) in human epidermal keratinocytes cultured on the fourth day.

Fig. 4A is a graph showing changes in the concentration of cholesterol caused by a DHCR7 inhibitor (AY-9944) in human epidermal keratinocytes on the first and fourth days after the start of differentiation. Fig. 4B is a graph showing the cellular respiratory activity caused by the DHCR7 inhibitor (AY-9944) in human epidermal keratinocytes on the first day after the initiation of differentiation.

Fig. 5 is a graph showing the gene expression of DHCR7 caused by catechin extract, MPC copolymer, retinyl acetate, and Winged Bean extract in human epidermal keratinocytes.

Fig. 6 is a graph showing the gene expression of an epidermal differentiation marker (case protein) caused by catechin extract in human epidermal keratinocytes.

Claims (4)

A cholesterol synthesis promoter comprising a group selected from the group consisting of catechin extract, a copolymer having 2-methylpropenyloxyethylphosphocholine, a retinyl acetate, and a tetragonal bean extract. At least one ingredient in the middle. A composition for promoting differentiation of epidermal cells, comprising the cholesterol synthesis promoter according to claim 1. A method for screening for an agent for promoting differentiation of epidermal cells, wherein when a candidate agent is applied to a cultured cell, an agent which promotes the expression of the DHCR7 gene of the cell is selected as a drug for promoting differentiation of epidermal cells. The method of claim 3, wherein the cultured cells are human epidermal keratinocytes.