KR101040477B1 - Composition for anti-androgenic effect - Google Patents

Abstract

The present invention provides an anti-androgen composition and a method for producing Ikaritin containing Ikaritin as an active ingredient.

The composition of the present invention exhibits anti-androgen activity and is effective for preventing, treating or ameliorating androgen-related diseases or conditions. In addition, it is possible to produce a high purity of Icarritin by the production method of the present invention.

Ikaritin, Antiandrogen

Description

Anti-androgen composition {Composition for anti-androgenic effect}

The present invention relates to an anti-androgen composition and to a method for producing ikaritin.

Androgens are male hormones and are known to be associated with diseases or conditions such as tumors of male hormone-dependent cells, androgenetic alopecia, acne (acne), adhesions (especially after abdominal surgery), and keloids.

Androgen's mechanism of action is known to pass through androgenic receptors (ARs), protein transcription factors that interact with specific regions of DNA. Thus, the mode of action of testosterone and its stronger homologue, 5-alpha dihydrotestosterone (DHT), relies on binding to the androgen receptor, and only after the androgen binds to the androgen receptor, transcription by the RNA polymerase II occurs. To show androgen action.

The anti-androgen drug refers to a drug that inhibits androgen action by blocking the receptor site activated by androgen so that androgen binding does not occur in the androgen receptor.

Anti-androgen drugs can be used to treat or alleviate androgen-related diseases or conditions.

For example, U. S. Patent No. 5,264, 619 discloses anti-androgenic monocyclic and morbid alkenes, which are caused by anti-androgen action, acne, androgenetic alopecia, infections of fungi with androgen receptors, Alzheimer's disease, osteoarthritis, breast capsules. It has been disclosed to have the effect of preventing and treating dysplasia (especially after breast formation), keloids, and adhesions.

In addition, Korean Patent No. 10-0508681 discloses an androgen receptor inhibitor, and has been shown to exhibit the effect of treating androgen syndrome, such as androgen-dependent tumors such as prostate cancer, acne, androgenetic alopecia by anti-androgen action. have.

In addition, the androgen receptor as putative therapeutic target in hormone-refractory prostate cancer. Recent Patents Anticancer Drug Discov. 2007 Nov; 2 (3): 203-11), and androgen-dependent tumors such as acne (Dermatology of androgen-related Clin Dermatol. 2006 Jul-Aug; 24 (4): 289-98), Androgen Alopecia (The importance of dual 5alpha-reductase inhibition in the treatment of male pattern hair loss: results of a randomized placebo-controlled study of dutasteride versus finasteride.J Am Acad Dermatol. 2006 Dec; 55 (6): 1014-23), and hirsutism; Treatment of hirsutism with combined pill containing drospirenone, Gynecol Endocrinol. 2008 Apr; 24 (4): 220-3 ), The antiandrogenic effect of flutamide improves uterine perfusion in women with polycystic ovary syndrome, Fertil Steril. 2002 Jun; 77 (6): 1136-40). Is disclosed.

Icarritin (icaritin) can be obtained by hydrolyzing Icarin in a form in which sugar is released from the glycoside Icarin (icariin). Ikariin is contained in the plant of the genus Epimedia, and when the epimedium is dried and used as a herbal medicine, it is known as yinyangquak as the herbal name.

Yin and Yang are the Epimedium brevicornum ), Epimedium sagittatum ), Epimedium pubescens , Epimedium wushanense or Epimedium koreanum , which is a dry ground area that strengthens the kidneys, strengthens tendons and bones, and relieves rheumatic symptoms. Although it is known, there is little research on the relationship with androgens.

In addition, Icarin, which is a glycoside component of Yin-Kak, has a problem in that it is difficult to obtain Icarritin by separating sugars by simple acid hydrolysis.

The technical problem to be solved by the present invention is to provide an anti-androgen composition.

In addition, the technical problem to be solved by the present invention is to provide a method for producing icaritin.

The present invention provides an anti-androgen composition containing icaritin as an active ingredient.

The present invention also provides a method for inhibiting androgen receptors comprising administering icaritin to a subject, and a method for inhibiting androgen receptors comprising administering a composition of the present invention to a subject.

The subject may be a mammal, including a human.

The present invention also provides anti-androgen uses of icaritin and anti-androgen uses of the compositions of the present invention.

Icarritin has anti-androgen activity, which is why Icartin is used for acne, androgenetic alopecia, prostate cancer, male hirsutism, polycystic ovary syndrome, infections of fungi with androgen receptors, Alzheimer's disease, osteoarthritis, breast encapsulation (especially breast plastic surgery). Post-occurring), keloids, and / or adhesions, and the like, for the treatment, improvement, and / or prevention of androgen-dependent diseases.

The composition of the present invention may be a pharmaceutical composition, a food composition or a cosmetic composition.

Ikaritin is a compound represented by the following Chemical Formula 1, and may be obtained by hydrolysis of Icarine, a compound represented by the following Chemical Formula 2, or synthesized. Icarine is Epimedium brevicornum ), Epimedium sagittatum ), Epimedium pubescens , Epimedium wushanense ) or Epimedium Correanum koreanum ) and the like can be used to extract from the plant of the epimediadium, etc. It can be used to extract from the medicinal herb with the name of Yinyangkak.

Ikaritin may be present in both solvated and unsolvated forms with pharmaceutically acceptable solvents such as water, ethanol and the like. In addition, the compound of Formula 1 may exist in different polymorphic forms, and the claims encompass all such forms.

The composition of the present invention may be a composition for the prevention and treatment of androgen-dependent diseases or conditions containing Ikaritin as an active ingredient.

The androgen-dependent diseases or conditions include acne, androgenetic alopecia, prostate cancer, male hirsutism, polycystic ovary syndrome, infections of fungi with androgen receptors, Alzheimer's disease, osteoarthritis, breast encapsulation (especially after breast formation), keloids (Keloids), and / or adhesions and the like.

The composition of the present invention can be formulated to include one or more pharmaceutical, food or / and cosmetically acceptable carriers in addition to the active ingredient for administration.

Pharmaceutical, food and / or cosmetically acceptable carriers may be used in combination with saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol and one or more of these components, If desired, other conventional additives such as antioxidants, buffers, bacteriostatics, and the like may be added. Diluents, dispersants, surfactants, solvents, disintegrants, sweeteners, binders, coatings, swelling agents, lubricants, lubricants or flavoring agents may additionally be added. Furthermore, it may be preferably formulated according to each disease or component by any suitable method in the art or using methods disclosed in Remington's Pharmaceutical Science (Recent Edition), Mack Publishing Company, Easton PA.

Formulation forms of the pharmaceutical compositions of the present invention include granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops or injectable solutions and sustained release formulations of the active compounds, and the like. This can be

The pharmaceutical composition of the present invention may be administered in a conventional manner through intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, nasal, inhalation, topical, rectal, oral, intraocular or intradermal routes, etc. May be administered.

In the pharmaceutical composition of the present invention, the dosage of the active ingredient may be adjusted within the range of 0.1 mg / kg to 500 mg / kg of icaritin based on an adult weight of 60 kg. However, the optimum dosage to be administered can be easily determined by those skilled in the art, and the type of disease, the severity of the disease, the amount of active ingredients and other ingredients contained in the composition, the type of formulation, and the age, weight, general health of the patient It may be adjusted according to various factors including the condition, sex and diet, time of administration, route of administration and the rate of secretion of the composition, the duration of treatment, and the drugs used simultaneously.

In addition, the food composition of the present invention may be for the prevention or improvement of androgen-dependent diseases or symptoms.

The androgen-dependent disease or condition may include acne, androgenetic alopecia, prostate cancer, male polymorphism, polycystic ovary syndrome, infections of fungi with androgen receptors, Alzheimer's disease, osteoarthritis, breast encapsulation (especially after breast plastic surgery), Keloids, and / or adhesions and the like.

The formulation of the food composition of the present invention may be prepared according to a conventional method, and may be encapsulated after drying with a carrier, or may be formulated in the form of other tablets, granules, powders, beverages, porridges, etc. It is possible to manufacture.

In addition, the cosmetic composition of the present invention may be for the prevention or improvement of androgen-dependent diseases or symptoms.

The androgen-dependent diseases or conditions include acne, androgenetic alopecia, prostate cancer, male hirsutism, polycystic ovary syndrome, infections of fungi with androgen receptors, Alzheimer's disease, osteoarthritis, breast encapsulation (especially after breast formation), keloids (Keloids), and / or adhesions and the like.

Formulation of the cosmetic composition of the present invention is prepared according to a conventional method, can be formulated in the form of a lotion, cream, etc. using a suitable carrier, in addition to the above-described can be prepared in all cosmetic forms.

The composition of the present invention may be administered topically. Topical administration refers to the direct application of icaritin (and the composition) to skin and / or hair. The topical compositions of the present invention may be in the form of solutions, lotions, waxes, creams, ointments, liposomes, sprays, gels, foams, roller sticks, or any other formulation commonly used in dermatology. have.

The composition of the present invention may also be composed of a solid preparation constituting a cleansing soap or bar, and may be prepared according to conventional manufacturing methods.

The compositions of the present invention can be used in the form of aqueous, alcoholic or aqueous-alcoholic solutions, hair, creams, gels, emulsions, mousses, aerosols for the hair. The compositions of the present invention may also be hair protection compositions, in particular shampoos, hair-setting lotions, treating lotions, styling creams or gels, dyeing compositions, lotions or gels for preventing hair loss, and the like.

The content of Ikaritin in the composition of the present invention may be contained in an amount of 0.001 to 99% by weight based on the total weight of the composition.

Ikaritin can increase the antiandrogen effect alone or in combination with topical minoxidil and its derivatives, which have been found to have an antiandrogen effect, and oral finasteride.

In addition, the present invention comprises the steps of (a) preparing an extract containing Icarin from a compound of the genus Epicardium containing Icarin using a solvent; (b) acid hydrolyzing the icaline containing extract; And (c) enzymatic hydrolysis of the acid hydrolyzate.

The step (b) may be to hydrolyze the kariin-containing extract with 3% sulfuric acid in 90% ethanol, the enzyme of step (c) may be beta-glucosidase.

Specifically, the method for producing Ikaritin of the present invention is as follows.

step (a) to obtain after filtration was repeated three times to extract heat the drying EK powder in 70% methanol was concentrated under reduced pressure methanol extract, suspending the methanol extract in water and CHCl _3, ethyl acetate, n-butanol (n-BuOH Fractions) sequentially. Ethyl acetate extract was used to obtain five small fractions (F1-F5) using silica gel column chromatography (ethyl acetate: methanol: water = 10: 2: 1). Separate phosphorus. Icarine-containing fractions were subjected to silica gel column chromatography (ethyl acetate: methanol: water = 100: 16.5: 13.5) to increase the purity of Icarine, followed by Sephadex LH-20 column chromatography. (MeOH)) can be used to obtain purified Ikariin.

Icarinine can be prepared from Icarin through steps (b) and (c), and the material obtained by mild acidic hydrolysis can be subjected to enzymatic hydrolysis, thereby making it. . The enzyme hydrolyzate was extracted and concentrated again with ethyl acetate to obtain an ethyl acetate extract, which was then subjected to silica gel column chromatography to obtain purified icaritin.

Icarine is difficult to obtain Ikaritin by separating sugars by simple acid hydrolysis method, and it is possible to prepare Ikaritin by mild acid hydrolysis and enzymatic hydrolysis of the present invention, and high purity by purification Preparation of tin is possible.

The composition containing the icaritin of the present invention exhibits anti-androgen action and is effective in preventing, treating or ameliorating androgen-related diseases or conditions. In addition, it is possible to produce a high purity of Icarritin by the production method of the present invention.

Examples are provided to help understand the present invention. The following examples are merely provided to more easily understand the present invention, but the contents of the present invention are not limited by the examples.

Example 1 Preparation of Icarritin

1) material

In May 2005, he visited the plantation in Cheorwon, Gangwon-do, and purchased 11 kg of the ground part of Epimedium koreanum Nakai (pharmaceutical name: Eumyanggok).

2) Icarine Separation from Yin Yang

end. Extraction and Solvent Fraction

-3.8kg of dry Yin-Yang powder was filtered through 70% methanol heating extraction three times, and concentrated under reduced pressure to obtain 970g of methanol extract.

The methanol extract was suspended in water and partitioned sequentially with CHCl ₃ , ethyl acetate, normal-butanol (n-BuOH).

I. Icarine Separation by Column Chromatography

60 g of ethyl acetate extract (61.3 g) was taken and five small fractions (F1-F5) were obtained using silica gel column chromatography (ethyl acetate: methanol: water = 10: 2: 1). Among them, fraction F3 (35.47 g) containing icarine was confirmed by silica gel column chromatography (ethyl acetate: methanol: water = 100: 16.5: 13.5) to increase the purity of Icarine, followed by Sephadex column. Chromatography (Sephadex LH-20 column chromatography (MeOH)) to obtain 4.8 g (yield 0.126%) of purified Ikari.

All. Identification and Purity Test

-Icarine was identified by direct comparison of standard with HPLC and ¹ H-NMR, ¹³ C-NMR, FAB-MS. HPLC was carried out under the following conditions to confirm the identification and purity by co-spotting with Ikariin standard (ChromaDex Inc.). HPLC chromatogram is shown in FIG.

HPLC conditions:

HPLC Pumps: Jasco 980 Series

Column: Atlantis C ₁₈ (4.6 × 150 mm)

Mobile phase: MeOH: H ₂ O (2: 1)

Flow rate: 1 mL / min

Injection volume: 10 μL

Detection: 230 nm

Purified Icarine is a yellow powder, and the formula is C ₃₃ H ₄₀ O ₁₅ , the molecular weight measured by LC-MS and FAB-MS is 676. HPLC showed 94.8% purity. (Preparative isolation and purification of three flavonoids from the Chinese medicinal plant Epimedium koreamum Nakai by high-speed counter-current chromatography. J Chromatogr A. 2005 Jan 28; 1064 (1): 53-7)

^-1 H-NMR (300 MHz, DMSO- d ₆ ) δ: 12.57 (1H, s , OH-5), 7.88 (2H, d , J = 8.7, H-2 ', 6'), 7.11 (2H, d , J = 8.7, H-3 ', 5'), 6.64 (1H, s , H-6), 5.14 (1H, d , J = 7.0, anomeric proton), 3.85 (3H, s , OCH ₃ ), 1.69 (3H, s , H-14), 1.60 (3H, s , H-15), 0.78 (3H, d , J = 5.7, H-6 ")

¹³ C-NMR (75 MHz, DMSO- d ₆ ) δ: 178.6 (C-4), 161.7 (C-7), 160.8 (C-5), 159.4 (C-4 '), 157.6 (C-9) , 153.3 (C-2), 134.9 (C-3), 131.4 (C-13), 130.9 (C-2 ', 6'), 122.5 (C-12), 122.4 (C-1 '), 114.4 ( C-3 ', 5'), 108.6 (C-8), 105.9 (C-10), 102.3 (C-1 "), 100.8 (C-1"'), 98.4 (C-6), 77.5 (C -3 "'), 76.9 (C-5"'), 73.7 (C-2 "'), 71.4 (C-4"'), 71.0 (C-3 "), 70.6 (C-2"), 70.4 (C-5 "), 60.9 (C-6"'), 55.8 (OCH ₃ ), 25.8 (C-14), 21.7 (C-11), 18.2 (C-6 "), 17.7 (C-15)

FAB-MS m / z : 677 [M + H] ⁺ , 531 [M + H-Rha] ⁺ , 369 [M + H-Rha-Glc] ⁺

4) Icarritin Isolation by Hydrolysis

The material obtained by mild acidic hydrolysis (3% sulfuric acid in 90% ethanol) was further subjected to enzymatic hydrolysis (beta-glucosidase, pH 5.0, overnight), followed by extraction and concentration with ethyl acetate. The ethyl acetate extract was obtained, and silica gel column chromatography (ethyl acetate: methanol = 5: 1) was used to obtain icaritin.

end. Mild Acid Hydrolysis

3% H ₂ SO ₄ in 90% ethanol in a round flask 100 mg of Ikari was added to 100 ml and refluxed in an oil bath for 90 minutes.

Add ice to stop the reaction immediately and evaporate under reduced pressure.

-Ethanol was blown out in a vacuum evaporator and water was extracted with the same amount of ethyl acetate.

The obtained ethyl acetate layer was washed 3-4 times with water to identify neutral pH paper.

The ethyl acetate layer was evaporated to give an acid hydrolyzate.

I. Enzyme hydrolysis

A and B solutions were prepared as follows, and A solution was slowly added while stirring B solution, and reacted for 24 hours at 37 ° C. in a shaking incubator.

Solution A: 50 mg of acid hydrolysate was dissolved in 45 ml of methanol.

B solution: 170 mg of beta-glucosidase was dissolved in 120 ml of 0.1 M acetic acid buffer (pH 5.0).

All. refine

Silica gel column chromatography (ethyl acetate: methanol = 5: 1) was carried out to obtain 1.3 g (yield 0.034%) of purified icaritin.

la. Identification and Purity Test

-Purified Ikaritin obtained through hydrolysis was identified through ¹ H, ¹³ C-NMR, EI-MS and purity confirmed by HPLC. HPLC chromatogram is shown in FIG. 2.

HPLC conditions:

HPLC Pumps: Jasco 980 Series

Column: Atlantis C ₁₈ (4.6 × 150 mm)

Mobile phase: acetonitrile (100%)

Flow rate: 1 mL / min

Injection volume: 10 μL

Detection: 230 nm

The chemical formula of the purified icaritin is C ₂₁ H ₂₀ O ₆ , the molecular weight measured by EI-MS was 368, and it was confirmed from the spectral data that the compound of formula 1 was icaritin, and it was confirmed that the purity was 97% by HPLC.

^-1 H-NMR (500 MHz, DMSO- d ₆ ) δ: 2.437, 2.557 (each 3H, s , C ₁₄ , ₁₅ -Me), 4.236 (2H, m , H-11), 4.652 (3H, s , OMe ), 5.985 (1H, brt, H-12), 6.29 (1H, s, H-6), 7.93 (2H, d, J = 11Hz, H-3 ', 5'), 8.93 (2H, d, J = 8.5 Hz, H-2 ', 6'), 10.25 (1H, s , C ₇ -OH), 11.53 (1H, s , C ₃ -OH), 13.16 (1H, s , C ₅ -OH)

¹³ C-NMR (500 MHz, DMSO- d ₆ ) δ: 185.878 (C-4), 170.972 (C-7), 170.129 (C-5), 167.951 (C-4 '), 163.179 (C-9) , 155.822 (C-2), 145.569 (C-3), 140.668 (C-13), 138.822 (C-2 ', 6'), 133.223 (C-1 '), 132.143 (C-12), 123.748 ( C-3 ', 5'), 115.284 (C-8), 112.691 (C-10), 107.492 (C-6), 65.036 (OCH ₃ ), 35.079 (C-14), 30.849 (C-11), 27.482 (C-15)

EI-MS ( m / z) 368 (100), 353 (78), 313 (59), 300 (44), 165 (11), 135 (33)

Example 2 Confirmation of Anti-androgen Effect of Ikari Tin Using PC3 / AR Cells

PC3 / AR cells transiently transfected androgen receptor (AR) plasmids (prepared by adding AR sequences to the pCMV plasmids) in PC3 cells (ATCC, USA) contain an androgen responsive element PMMTV-Luc plasmid (prepared by inserting MMTV promoter into pGL3 basic vector) and plasmid expressing Renilla luciferase to measure transfection efficiency (pRL vector, promega, PC3 / AR cells were prepared by co-transfection (using Exgene 500 kit, MBI Fermentas).

In the prepared PC3 / AR cells, 5-alpha dihydrotestosterone (DHT) (Wako, Japan) or / and the icaritin of Example 1 were treated for 48 hours at the concentrations of Table 1, followed by dual activity of luciferase expressed. Measured with Luciferase Assay Kit (Promega, USA).

division DHT Icarine DHT treatment 1 × 10 ^-8 M - DHT / icaritin treatment (1 × 10 ^-6 M) 1 × 10 ^-8 M 1 × 10 ^-6 M DHT / icaritin treatment (5 × 10 ^-6 M) 1 × 10 ^-8 M 5 × 10 ^-6 M DHT / icaritin treatment (10 × 10 ^-6 M) 1 × 10 ^-8 M 10 × 10 ^-6 M DHT / icaritin treatment (20 × 10 ^-6 M) 1 × 10 ^-8 M 20 × 10 ^-6 M DHT / icaritin treatment (50 × 10 ^-6 M) 1 × 10 ^-8 M 50 × 10 ^-6 M

Relative luciferase activity results are shown in FIG. 3. NT in Figure 3 refers to the control group that did not have any treatment. As a result, it was confirmed that the icaritin shows anti-androgen activity in PC3 / AR cells in a concentration-dependent manner, compared to the case where the ikaritin 50 × 10 ⁻⁶ M treated PC3 / AR cells of Example 1 were not treated with icaritin. It has been shown to inhibit the action of 5-alpha dihydrotestosterone (DHT) by 53.8%.

Example 3 Confirmation of Anti-androgen Effect of Icarritin-Experimental Inhibition of Androgen and Androgen Receptor Binding by Ikaritin

Effect of Icartin on androgen and androgen receptor binding was confirmed by Kim et al. (Effects of 2,4-D and DCP on the DHT-Induced Androgenic Action in Human Prostate Cancer Cells, 2005, TOXICOLOGICAL SCIENCES 88 (1), 52-59) It was carried out according to the method.

CHO / AR cells (prepared by transfection of pCMV-AR in cell lines purchased from ATCC) were prepared.

5-alpha dehydrotestosterone (DHT) (Wako, Japan) and Ikaritin, each labeled [ ³ H], were treated at the concentrations shown in the following table and left for 2 hours, and the cells were treated with phosphate buffered saline. After washing twice, cells were harvested with buffer containing pH 2, SDS, 10% glycerol and 10 mM Tris (pH 6.8), followed by measurement of radiation dose to determine 5-alpha dehydrotestosterone (DHT) labeled [ ³ H]. ) And the androgen receptor binding rate was measured. The results are shown in Fig. 4 (▲). Increasing the concentration of Ikaritin was shown to decrease the binding rate with androgen receptor of 5-alpha dihydrotestosterone (DHT) labeled [ ³ H].

division DHT Icarine Ikaritin untreated group 5 nM - Ikaritin treatment group (5nM) 5 nM 5 nM Ikaritin treatment group (50 nM) 5 nM 50 nM Icarritin Treatment Group (500nM) 5 nM 500 nM

In addition, 5-alpha dihydrotestosterone (DHT) (Wako, Japan) and unlabeled 5-alpha dihydrotestosterone (DHT) labeled [ ³ H] were prepared at the concentrations described in the following table, respectively. In the same manner as in the case of the measurement, the radiation dose was measured to measure the binding rate of 5-alpha dehydrotestosterone (DHT) and androgen receptor labeled with [ ³ H]. The results are shown in Figure 4 (◆). Increasing concentrations of unlabeled 5-alpha dihydrotestosterone (DHT) have been shown to decrease the binding rate with androgen receptors of 5-alpha dehydrotestosterone (DHT) labeled [ ³ H].

division DHT ^[3 H] labeled DHT [ ³ H] no cover DHT 5 nM - ^[3 H] labeled DHT treatment (5nM) 5 nM 5 nM ^[3 H] labeled DHT treatment (50nM) 5 nM 50 nM ^[3 H] labeled DHT treatment (500nM) 5 nM 500 nM

It can be seen from the above results that the binding of androgen and androgen receptors is inhibited by icaritin, and these results indicate that the mechanism of anti-androgen action of icaritin may be due to inhibition of the binding of androgen and androgen receptors.

Example 4 Confirmation of Anti-androgen Effect of Icarritin-Inhibition of Nuclear Migration of Androgen Receptor Bound to Androgen of Ikaritin

PC3 cells transiently expressing pAR-GFP (PC3 cells (ATCC) were transfected with a plasmid cloned with AR in a GFP vector (Invitrogen)).

GFP-AR migrates to the nucleus after 30 minutes after treatment with the pary-GFP-expressing PC3 cells with the karitin and / or 5-alpha dihydrotestosterone (DHT) of Example 1 at the concentrations shown in the following table. Aspects were observed by confocal microscopy (LSM 510 META, ZEISS). The results are shown in Fig. The control of FIG. 5 is an observation result for pAR-GFP-expressing PC3 cells not treated with icaritin and 5-alpha dihydrotestosterone (DHT).

Ikaritin concentration DHT concentration Icarritin Treatment 1 × 10 ^-6 M - Icarine / DHT Treatment 1 × 10 ^-6 M 1 × 10 ^-9 M DHT treatment - 1 × 10 ^-9 M

AR-GFP did not move into the nucleus when treated with Ikaritin, and AR-GFP did not move into the nucleus when co-treated with Ikaritin and 5-alpha dihydrotestosterone (DHT).

In contrast, the treatment of 5-alpha dehydrotestosterone (DHT) caused the AR-GPF of many cells to migrate into the nucleus.

From the above results, the treatment of andrigen does not move to the nucleus and thus no androgen action by the treatment of icaritin alone, and Ikaritin inhibits the transfer of the androgen receptor bound to 5-alpha dihydrotestosterone (DHT) into the nucleus. see.

Androgens bind to androgen receptors, move into the nucleus, and then act as an androgen responsive element to show androgen action, thus moving to the nucleus of 5-alpha dehydrotestosterone (DHT) bound androgen receptors. This inhibition results in anti-androgen action.

From the results of Examples 2 to 4, icaritin seems to exhibit anti-androgen action by mechanisms such as inhibition of androgen binding to the receptor and inhibition of transfer of androgen bound to the receptor into the nucleus.

Acne, androgenetic alopecia, prostate cancer, male hirsutism, polycystic ovary syndrome, infection of fungi with androgen receptors, Alzheimer's disease, osteoarthritis, breast encapsulation (especially after breast formation) by the anti-androgen action of the kariritin Androgen-dependent diseases such as, keloids, or adhesions can be prevented and treated.

Figure 1 shows an HPLC chromatogram of Icarine prepared according to one embodiment of the production method of the present invention.

Figure 2 shows the HPLC chromatogram of Icarritin prepared according to one embodiment of the production method of the present invention.

Figure 3 is a graph showing the results of luciferase activity measurement for measuring the anti-androgen effect of kariritin in PC3 / AR cells.

Figure 4 is a graph showing the results of measuring the effect of Ikari on the binding of 5-alpha dihydrotestosterone (DHT) and androgen receptor.

Figure 5 is a diagram showing the results confirming the nuclear transfer inhibitory effect of the androgen-bound androgen receptor of the kariritin.

Claims (16)

A pharmaceutical composition for anti-androgens containing Ikaritin as an active ingredient. According to claim 1, wherein the composition is a composition for the treatment or prevention of androgen-dependent diseases or conditions. According to claim 2, The androgen-dependent disease or condition is acne, androgenetic alopecia, prostate cancer, male hirsutism, polycystic ovary syndrome, infection of fungi with androgen receptor, Alzheimer's disease, osteoarthritis, breast encapsulation, keloids (Keloids) ), And adhesions. The composition according to any one of claims 1 to 3, further comprising a pharmaceutically acceptable carrier. The composition according to any one of claims 1 to 3, wherein the icaritin is obtained by hydrolyzing icaline extracted from Yin and Yang. The composition according to any one of claims 1 to 3, wherein the icaritin is obtained by hydrolyzing icaline extracted from an epimedium plant. The method of claim 6, wherein the genus Epimedium plants are Epimedium brevicornum, Epimedium sagittatum, Epimedium pubescens, Epimedium wushanense and At least one composition selected from Epimedium koreanum. delete delete delete delete An anti-androgen food composition for the prevention or amelioration of androgen-dependent diseases or symptoms containing icartin as an active ingredient. An anti-androgen cosmetic composition for the prevention or amelioration of androgen-dependent diseases or conditions containing Ikaritin as an active ingredient. delete The method of claim 12, wherein the androgen-dependent disease or condition is acne, androgenetic alopecia, prostate cancer, male hirsutism, polycystic ovary syndrome, infection of fungi with androgen receptor, Alzheimer's disease, osteoarthritis, breast encapsulation, keloids (Keloids) ), And adhesions. According to claim 13, The androgen-dependent disease or condition is acne, androgenetic alopecia, prostate cancer, male hirsutism, polycystic ovary syndrome, infection of fungi with androgen receptor, Alzheimer's disease, osteoarthritis, breast encapsulation, keloids (Keloids) ), And adhesions.

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