Methods for the evaluation or selection of sebaceous gland or hair follicle selective androgen receptor activity controllers
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一种雄激素受体、皮脂腺的技术，应用在活性进行控制的物质领域，能够解决关系未知等问题
Active Publication Date: 2021-06-25
KAO CORP
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However, the relationship between androgen receptors in sebaceous glands and hair follicles, HIF1α or glycolytic enzymes is unknown
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[0192] Hereinafter, an Example is shown and this invention is demonstrated more concretely.
[0193]
reference example 1
[0194] Reference Example 1. Cells
[0195] Human sebocyte cell line (SZ95) was cultured in Sebomed containing 10% FBS, 5ng / mL epidermal growth factor (EpidermalGrowth Factor) TM Cultured in basal medium (Biochrom). A human prostate cancer cell line (LNCaP) was cultured in RPMI1640 (Life technologies) containing 10% FBS. Usually, cultivation under oxygen concentration is carried out in air (oxygen concentration 20.9%). For culture under hypoxic conditions, AnaeroPack-Anaero 5% (Mitsubishi Gas Chemical Co., Ltd.) and an angled wide-mouth thermos flask (SUGIYAMA-GEN CO., LTD) were used in an oxygen concentration of 0.1%, 5% CO 2 and cultured at 37°C. SZ95 was supplied by Prof. Dr. Prof. h.c. Dr. h.c. Christos C. Zouboulis. LNCaP was purchased by ATCC.
reference example 2
[0196] Reference Example 2. Preparation of Plasmid DNA
[0197] (1) Androgen receptor gene
[0198] Using PrimeSTAR (registered trademark) GXL DNA Polymerase (Takara bio), the open reading frame (ORF) region of the gene encoding human androgen receptor (androgen receptor, AR) was amplified by PCR, followed by In-FusionHD Cloning Kit ( Takara Bio) was inserted into pcDNA3.1 (Lifetechnologies) treated with EcoRI. From Escherichia coli transformed with the plasmid DNA of interest, the plasmid DNA was purified using the EndoFree Plasmid Purification Kit (Qiagen). The purified plasmid DNA was named pcDNA3.1-AR.
[0199] (2) Glycolytic enzyme gene
[0200] Enolase 1, lactate dehydrogenase A, glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase 1, aldolase A, Insertion of the open reading frame (ORF) region of the genes for hexokinase 1, glucose-6-phosphate isomerase, and triosephosphate isomerase (ENO1, LDHA, GAPDH, PGK1, ALDOA, HK1, GPI, and TPI, respectively) to ...
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Abstract
The present invention provides a method for searching for a substance for rapidly, accurately, and selectively controlling the activity of androgen receptors in sebaceous glands and hair follicles. A method for evaluating or selecting a sebaceous gland or hair follicle selective androgen receptor activity controller, comprising: (A) applying a substance to be tested to cells derived from sebaceous glands or cells derived from hair follicles under hypoxic conditions; B) measuring the activity of HIF1α in the cell; (C) comparing the activity determined in (B) with the activity of HIF1α in the control group; and (D) evaluating the substance to be tested based on the result of (C) The effect of controlling the activity of HIF1α.
Description
technical field [0001] The present invention relates to a method for finding substances for selectively controlling the activity of androgen receptors of sebaceous glands or hair follicles. Background technique [0002] The androgen receptor (AR) is a receptor for steroid hormones including the mineralocorticoid receptor (MR), progesterone receptor (PR), estrogen receptor (ER), and glucocorticoid receptor (GR). Part of the body subfamily. Endogenous steroidal androgens (eg, testosterone and 5α-dihydrotestosterone (DHT)) are the major circulating sex hormones that play important roles in the regulation of various physiological processes in addition to promoting secondary sexual characteristics. Regarding the effect of androgens on the skin, oral administration of dehydroepiandrosterone (DHEA) has been reported to improve skin condition in the elderly; and topical administration of DHEA to the skin, usually in postmenopausal women with reduced sebum volume The amount of midd...
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