KR100310613B1 - Euphorbia Lathyris extract inhibiting the activity of 5a-reductase, and the cosmetic or skincare-pharmaceutical composition thereof for suppressing pimple or sebum - Google Patents

Abstract

The present invention relates to a 5α-reductase inhibitor, characterized in that it contains a water extract isolated from methanol or ethanol. In addition, the present invention relates to a method of using a fastener extracted with methanol or ethanol to suppress the side effects caused by dihydrotestosterone produced by 5α-reductase. Furthermore, the present invention relates to a sebum inhibiting or acne treatment cosmetics or skin coatings containing the extract from methanol or ethanol.

The water extract is not only excellent in 5α-reductase inhibitory effect, but also excellent in acne relief and sebum secretion when used as a cosmetic composition in humans. Therefore, the water extract according to the present invention has an acne laxative effect or sebum secretion inhibitory effect at a concentration that does not affect the safety of the product, so it can be used as an effective concentration in sebum inhibitory cosmetics or acne laxative cosmetics, and general dosage forms of medicines for skin application It can be used as an active ingredient in.

Description

Euphorbia Lathyris extract inhibiting the activity of 5a-reductase, and the cosmetic or skincare-pharmaceutical composition about for suppressing the acne extract that inhibits the activity of alpha-reductase, and acne or sebum inhibitor containing the same pimple or sebum}

The present invention relates to a 5α-reductase inhibitor, characterized in that it contains a water extract isolated from methanol or ethanol.

In addition, the present invention relates to a method of using a fastener extracted with methanol or ethanol to suppress the side effects caused by dihydrotestosterone produced by 5α-reductase.

Furthermore, the present invention relates to a sebum inhibiting or acne treatment cosmetics or skin coatings containing the extract from methanol or ethanol.

5α-reductase is present in testosterone-reactive tissues such as sebaceous glands, hair follicles, prostate, and epididymis, and reduces testosterone, one of the testosterones, to dihydrotestosterone (DHT). As an enzyme involved, its conversion requires NDPH (NADPH). Testosterone is involved in male sexual impulses, skeletal muscle growth, male external genitalia, scrotum growth, and spermatogenesis, while dihydrotestosterone is involved in acne, sebum and prostatic hyperplasia (Diane et al; JID 775-778, 1995. Bruchovsky, N et al; JBC 243, 2112-2121, 1968). Therefore, studies are actively underway to develop acne treatment and sebum secretion inhibitors using inhibitors of 5α-reductase enzyme.

In general, sebum in the skin, including the scalp and face, plays a role in moisturizing the skin and preventing invasion of microorganisms, but promotes acne after puberty. In particular, women in their 30s and older may cause skin bloating, lifting of makeup, and widening of pores due to excessive secretion of sebum. Sebaceous hypersecretion is caused by a number of causes, the most important of which is the sebaceous gland is activated by the amount of dihydrotestosterone, one of the hormones involved in promoting sebum secretion, sebum hypersecretion It is known to happen.

Acne is a complex cause of various causes, such as pubertal hyperactivity of the sebaceous glands secrete vigorously, epithelial cells of the hair follicles cause dyskerosis, clogging the hair follicles, which forms the scleroderma, which is the primary lesion of acne. Bacteria that reside in hair follicles produce free fatty acids, which stimulate hair follicles to cause inflammation. This progression to acne is exacerbated by sebum oversecretion.

For the treatment of acne caused by this testosterone suppression, female hormones such as estrogen or the like have been used or 5α-reductase inhibitors. However, such female hormones, in addition to the acne treatment effect, skin inflammation, side effects caused by the hormone has been reported, and the current situation is discontinued or use a very small amount. On the other hand, as an acne treatment agent that inhibits sebum oversecretion, it is commercially available as a formulation containing 15-20 excess of azelaic acid, an inhibitor of 5α-reductase.

On the other hand, even in the cosmetic field, sebum oversecretion is an important subject of interest as a factor that causes the lifting of the makeup and widening of the pores, the research is insignificant. Currently, sebum inhibitory cosmetics use a porous powder that temporarily adsorbs sebum or ivy extract, which is a herbal extract known to inhibit sebum, as a cosmetic raw material, but its effect is minimal.

The present invention, in view of this reality as part of the research to overcome the problems of the known sebum inhibitory substance was searched for an active substance among natural products, to find an extract of natural products exhibiting the inhibitory effect of 5α-reductase. To determine whether these natural extracts have an acne reduction or sebum secretion effect.

The present invention relates to a 5α-reductase inhibitor, characterized in that it contains a water extract isolated from methanol or ethanol.

In addition, the present invention relates to a method of using a fastener extracted with methanol or ethanol to suppress the side effects caused by dihydrotestosterone produced by 5α-reductase.

Furthermore, the present invention relates to a sebum inhibiting or acne treatment cosmetics or skin coatings containing the extract from methanol or ethanol.

The present inventors have tried to find a natural extract that inhibits the activity of 5α-reductase, an enzyme catalyzing the production of dihydrotestosterone, in order to control the amount of dihydrotestosterone known to cause sebum secretion and acne production.

Using 300 extracts of several natural plants, including Succulents, to determine whether they inhibit the activity of 5α-Reductase in vitro, we found that Succulent Extracts act as inhibitors of 5α-Reductase. Found (see Experiment 1).

On the basis of this, the present inventors have found that the water extract is showing an excellent effect as a result of acne reduction and sebum secretion using the water extracts [see Experiments 2 to 4].

Therefore, the present invention intends to use the extracts isolated from methanol or ethanol to inhibit the activity of 5α-reductase, and furthermore, the extracts are used to reduce acne and sebum secretion.

It will be described in detail with respect to the water extract according to the present invention.

The abbreviation (Euphorbiae Lathyridis Semen; botanical name: Euphorbia Lathyris L), also known as sperm, larva or white, is a biennial vegetation, grows about 1m straight, has a willow leaf shape, has many branches, and is succulent. The plant blossoms in April-July, the fruit is July-August, the seeds are elliptical, 5-6 mm long, 4 mm in diameter and glossy on the inside. Since ancient times, the deceased has the effect on the long bowel, hepatic pluripotency, dysplasia, menstruation, food poisoning, and poisonous insects. The main known ingredients include oils 40-48, coumarin-based daphrine and esculletin 0.6, bicoumarin-like substances eupol, α-euphorball and isoeupo, similar to Phadu's diarrhea. Triterpenoids such as betin and isoeuphorin (the ingredients and their use in herbal medicine, Jangwolseo, 1991).

According to the present invention, a method of extracting and concentrating a water extract of water separated by methanol or ethanol is described in detail as follows.

After crushing the natural product or the herbal medicine, the active ingredient is extracted from the natural water by using ethanol or methanol which is anhydrous lower alcohol as an extraction solvent.

In order to maximize the elution component, it is preferable to use methanol as the extraction solvent, which can destroy the cell membrane.

In order to prevent the effective components from evaporating, it is preferable to extract by heating in the appropriate temperature range of each extraction solvent in the state that the cooling capacitor is equipped.

The titration temperature of methanol is 50-80 캜, and the titration temperature of ethanol is 65 캜.

On the other hand, it is preferable to extract by heating for 2 hours in order to maximize the amount of elution without destroying the active ingredient.

It is preferable to remove the impurities by adding the extraction solvent to the inner water and leaving the separated extract at room temperature for one day to precipitate impurities, and then filtering with a wattman filter paper. At this time, the extract is clear and yellow and green in appearance.

To evaporate the solvent, the solvent is recovered by evaporation using a distillation apparatus. At this time, it is preferable to use a distillation apparatus with a cooling capacitor in order to prevent the active ingredients of the extract from evaporating.

In order to further concentrate the water extract, the water extract is mixed with a solvent consisting of water and ethyl acetate or a solvent consisting of water and butanol, and dissolving both water-soluble and non-aqueous components of the water eluted by the extraction solvent, After separating the layers, the ethyl acetate layer or the butanol layer is recovered.

In the fractionation, the excess of the active ingredient is detected in the ethyl acetate layer or the butanol layer of the supernatant rather than the water layer, and the supernatant is separated to recover the ethyl acetate or butanol that is evaporated using a distillation apparatus equipped with a cooling capacitor, and the concentrated liquid is concentrated under reduced pressure. Obtain an aqueous solution of. When applying water extract to cosmetics, ethyl acetate or butanol should be completely removed to avoid odor.

In order to remove the wax deposited on the extract, it is preferable to remove the wax by filtration in vacuo using an edbenten-like filter paper and to separate only the liquid phase by penetrating only the liquid phase. This is because the addition of water-soluble extract in the cosmetics may cause problems in the formulation if a lot of wax is left.

The water extract isolated or further concentrated in the above manner is dissolved in 501,3-butylene glycol and 10 ethanol, and can be used by adding to cosmetics such as softening water, nutrient cosmetics, massage cream, essence, pack, and the like. In addition, the extract may be used as an active ingredient in the general formulation of the drug for skin application.

Hereinafter, the present invention will be described in detail through examples and experiments. However, the scope of the present invention is not limited by the above examples and experiments.

[Example 1; Preparation of Succulent Extracts]

(1) Fastener natural product is pulverized, and added methanol to dry weight (kg) in a ratio of 1: 1 (w / v), and then heated at 65 ° C. for 2 hours with a cooling capacitor. By extracting the active ingredient of the recipient. The primary extract from this was stored.

(2) Methanol was added to the dry weight (kg) of the fastener natural product (1) from which the primary extract was removed in a ratio of 1: 1 (w / v), and in the state where the cooling capacitor was installed (1) ) Process was repeated, extracting the active ingredients of the beneficiary as much as possible (secondary extract).

(3) After mixing the primary extract and the secondary extract, it was left at room temperature for one day to precipitate impurities, and then filtered twice with No. 2 filter paper Whatman to completely remove impurities. At this time, the extract was shown to be a clear yellow to green intermediate.

(4) The extract obtained in (3) was evaporated using a distillation apparatus equipped with a cooling capacitor to recover the solvent from it, and the extract was completely concentrated under reduced pressure.

(5) To the depressurized extract, 3 volumes of a solvent mixed with water and ethyl acetate in a 1: 1 ratio were added to complete turbidity. This turbidity dissolved both the water-soluble and water-insoluble components eluted with methanol and left for one day to separate the layers. The ethyl acetate layer as a supernatant was separated, and the ethyl acetate extracted by evaporation was recovered using a distillation apparatus equipped with a cooling capacitor, to obtain a rapid depressurized liquid concentrate completely concentrated under reduced pressure.

(6) The fastener concentrate was filtered in vacuo using an adbenten-like filter paper to remove wax, and only the permeate liquid phase was separated.

Finally, 0.05 to 0.2 of the aqueous extract was obtained (Sample 1).

[Example 2; Preparation of Succulent Extracts]

70 ethanol was added to the dry weight (kg) of the fastener natural product in a ratio of 1: 1 (w / v), and the active ingredient of the fastener was extracted by heating at 65 ° C. for 2 hours while the cooling capacitor was installed. The fastener extract was evaporated using a distillation apparatus equipped with a cooling capacitor to recover the solvent from the fastener extract, thereby obtaining a concentrated extractor which was completely concentrated under reduced pressure.

The mixture is then mixed with ethyl acetate and water in a ratio of 1: 1, and the layers are separated, and only the portion of the ethyl acetate layer is taken out and concentrated under reduced pressure while recovering the ethyl acetate which is evaporated using a distillation apparatus equipped with a cooling capacitor. A water extract of the prepared liquid was obtained. The adjuvant concentrate was filtered in vacuo using an adbenten-like filter paper to remove the wax, and only the permeated liquid phase was separated.

This resulted in 0.3-0.5 fastener concentrate (Sample 2).

[Example 3; Preparation of Succulent Extracts]

50ethanol was added 1: 1 (w / v) to the dry weight of the fastener natural product (kg), and the active ingredient of the fastener was extracted by heating at 65 ° C. for 2 hours while the cooling capacitor was installed. The fastener extract was evaporated using a distillation apparatus equipped with a cooling capacitor to recover the solvent from the fastener extract, thereby obtaining a concentrated extractor which was completely concentrated under reduced pressure.

This was again mixed with non-polar butanol and water more than ethyl acetate in a ratio of 1: 1, and the layers were separated, and only the part of the butanol layer was taken out and recovered by evaporation of butanol using a distillation apparatus equipped with a cooling capacitor. A concentrated aqueous solution was obtained. The adjuvant concentrate was filtered in vacuo using an adbenten-like filter paper to remove the wax, and only the permeated liquid phase was separated.

From this, a water concentrate of 0.1-0.3 was obtained (sample 3).

[Example 4; Soft Cosmetics]

Sebum inhibitory nutrition lotion containing the extract of Example 1 (Sample 1) was prepared according to the formulation example of Table 1.

(Part by weight) Sucrose extract 0.5 beeswax 4.0 polysorbate 60 1.5 sorbitan sesquiolade 0.7 fluid paraffin 5.0 squalane 5.0 caprylic / capric triglyceride 5.0 glycerin 3.0 butylene glycol 3.0 propylene glycol 3.0 carboxyvinyl polymer 0.1 triethanolamine 0.2 Preservative proper pigment proper amount Total 100.0

Experiment 1; 5α-Reductase Inhibitory Effect Test of Sokja Extract>

In order to confirm whether the extracts are inhibitors that inhibit the activity of 5α-reductase, the following experiment was performed.

1) A male Sprogue-Dawley rat (7-8 weeks of age) was killed with a deciduous rat, and then 0.32M sucrose in which the abdominal prostate was removed to remove connective tissue. A buffer containing 0.1 mM dithiothreitol, 20 mM sodium acetate, was added, finely chopped and suspended in a grinder. After centrifugation of the suspension, the supernatant was taken. 5α-reductase is partially purified in this supernatant.

2) A portion of the supernatant obtained above was taken and placed in a buffer containing 0.2M monobasic and 0.2M dibasic.

3) pH 6.5, 1mM hit by a die thio tray tolyl, ADP 40mM sodium phosphate (Sodium Phosphate), 50μM yen ^{(NADPH), 2.2 × 10 9} M [1,2,6,7- 3 H] testosterone / testosterone 565 µl of the reaction solution was prepared.

4) The water extracts (samples 1 to 3) obtained in Examples 1 to 3 were dissolved in 10 ethanol to 0.1, respectively, and then 5.65 µl was added to the reaction solution of 3) to obtain a final concentration of 0.001.

As a control, 5.65 [mu] l of 10 ethanol without the addition of the extract was added to 565 [mu] l of the reaction solution of step 3).

5) As soon as 0.8 mg of the enzyme suspension of 2) partially purified in 1) was finally added, the reaction proceeded at 37 ° C. for 30 minutes.

6) Then, when 1 ml of ethyl acetate is added and clouded, the isotopic testosterone and dihydrotestosterone are separated from the supernatant. An ethyl acetate phase of 100 µl of the supernatant was developed on silica plastic sheet kieselgel 60 F254 using the developing solvent system as ethyl acetate-cyclonucleic acid (1: 1). Through this process, dihydrotestosterone converted from testosterone by 5-α reductase was separated from testosterone by the difference in distance.

After drying the plastic sheet in air, the bath system was used to measure the amount of isotopes. That is, the dried plastic sheet and the X-ray film were put together in the bath cassette and stored for one week, and then the amount of isotopes of testosterone and dihydrotestosterone remaining in the film was measured.

7) The experiment was repeated two times.

8) 5α- deokte Li order to measure the rise activity was measured generation amount of testosterone substrate ^[3 H] dihydro testosterone ^[3 H] from.

Based on the measurement data, the results for the inhibition rate of 5α-reductase activity in each of Samples 1 to 3 are shown in Table 2. In order to remove the errors caused by the natural degradation of impurities or substrates, the substrate control values without enzymes were subtracted from the results of the enzyme inhibition activity screening.

Experimental Results of 5α-Reductase Activity Inhibition by Influx Extracts T (dpm) DHT (dpm) Conversion rate Inhibition rate () Primary Sample 1 (Example 1) 10920.0 945.8 6.3 87.8 Sample 2 (Example 2) 12990.8 2612.1 15.1 70.8 Sample 3 (Example 3) 12762.0 2146.5 12.7 75.4 Control 7506.9 8572.3 53.8 - Secondary Sample 1 (Example 1) 12671.8 1230.0 7.4 70.5 Sample 2 (Example 2) 11820.0 1737.7 11.3 56.0 Sample 3 (Example 3) 12350.9 1733.2 10.8 58.6 Control 9708.0 3839.2 26.7 -

* T: ³ H radio activity in the testosterone region

DHT: ³ H radioactivity in the dihydrotestosterone region

* Conversion rate: Radioactivity / total radiation in DHT area

Inhibition Rate (): 100 × (Conversion Rate-Sample Conversion Rate) / Control Rate

As can be seen from the above results, the extracts (samples 1 to 3) extracted by Examples 1, 2, and 3 inhibit the activity of 5α-reductase, and are dies which are male hormones related to 5α-reductase. It can be seen that there is an effect of suppressing the amount of hydrotestosterone produced. In addition, it can be seen that the extract of Example 1 (sample 1) has the most inhibitory effect on the amount of production of dihydrotestosterone, a male hormone associated with 5α-reductase.

Experiment 2; Cotton Inhibition Test>

Treating a cotton-causing agent in the rabbit's ears will cause the cotton to grow after one week, but the larger cotton usually looks the same as before acne's inflammation. As the cotton cloth grows, there is an excessive amount of sebum in the pores, which causes the pores to be wider than normal. Therefore, the scleroderma experiment is somewhat different from the acne effect in humans, but at present it is the closest experimental method that can induce wide pores.

In order to determine whether the extract of Soukja extract (sample 1) extracted in Example 1 was effective in suppressing cotton cloth, the following experiment was performed.

1) 150 μl of 50-oleic acid / paraffin oil, which is a cotton-causing substance, is applied to the ears of white rabbits once a day to induce cotton cloth. 1, dissolved in 1.3-BG, 0.2 ml twice was applied in the afternoon.

2) As a control group, the group was treated with oleic acid, which was not treated with the oleic acid (control group 1), and treated with oleic acid, and the extract was treated with the group that was not applied (control group 2).

3) The number of cotton cloth was based on the size of the cotton cloth being 0.2 mm or more.

4) After exfoliation of the tissues of the scalp generating site by 0.8 mm, the keratin was removed, and the size of the scalp was measured by an optical microscope. # 1-6 is to measure the size of the cotton cloth by randomly selecting six of the well-developed cotton cloths from the sliced tissue. The results are shown in Table 3.

Measuring the Number and Size of Cotton Cloth Using the Ears of Earthenware Cotton catcher Cotton cloth size measurement (0.1mm) #One #2 # 3 #4 # 5 # 6 Average Not treated with oleic acid (control 1) N = 2 0 1.5 2 2 2 2 2 1.78 1.5 1.5 2 2 1.5 1.5 Oleic Acid Treatment (Control 2) N = 3 13 4 5 6 4 5 4 4.38 5 5 4 4 4 3 4 5 4 4 5 4 Oleic acid treatment + water extract application (Example 1) N = 3 5 3 3 2 4 2 3 2.83 3 3 3 3 3 2 3 3 3 2 3 3

N: number of animals

# 1-6: Serial number of six cotton cloths selected for measuring cotton size

In animal experiments with rabbits, the number and size of cotton cloth gradually decreased from 2 weeks after application of the extracts.

In addition, as can be seen in Table 3, it can be confirmed that the extracts according to the present invention has an effect of suppressing the generation of cotton cloth. In other words, the reduction in the number and size of cotton cloth can be inferred from the effects on acne, as well as the reduction in pore size in people with enlarged pores due to sebum oversecretion.

Experiment 3; Acne Effect>

Ten men and women who suffer from acne (Table 4) were selected and softened according to the prescription of Example 4, and the sebum secretion of the acne and facial lotion for 4 weeks in the morning and evening were corrected daily. Judgment of the effect was scored according to the degree felt by the person who participated in the test.

The experimental results are shown in Table 4.

Acne Relief Measure Pre-test status Effect (after 2 weeks) Effect (after 4 weeks) Person 1 One - + Person 2 3 ++ ++ Person 3 2 + ++ Person 4 One + + Person 5 2 - + Person 6 3 - ++ Person 7 2 ++ +++ Person 8 One + ++ Person 9 2 + + Person 10 2 + ++

Pre-test status: 1 = slight acne

2 = slightly

3 = severe acne

Effectiveness judgment:-: Almost no effect

+: Has some effect

++: eased a lot

+++: completely healed

Experiment 4; Sebum Secretion Suppression>

The following experiment was conducted on six men and women who felt sebum secretion on the face to determine whether and how the extracts inhibit sebum secretion.

1) After washing the face every evening, the flexible longevity of Example 4 was hoped for a month.

2) Sebum measurement was stabilized in a constant temperature and humidity room for 30 minutes after washing the face, and then sebum was measured at the three parts of the forehead, cheek, and chin by using a sebumeter.

The amount of sebum measured before use of the extract was set to the initial value. Sebum amount was measured once a week for 4 weeks while continuously using the softening water (Example 4) containing a water extract.

3) Compared with the initial value, it was determined whether sebum inhibiting efficacy of softener longevity containing water extract is shown in Table 5.

Sebum secretion suppression Sebum amount (mg / cm ² h) forehead cheek chin Initial value 92.17 ± 26.35 75.17 ± 12.94 87.33 ± 35.05 Week 1 81.67 ± 22.49 56.5 ± 9.75 77.17 ± 23.66 2nd week 83.67 ± 13.35 73.33 ± 12.91 84.17 ± 33.77 3rd week 81.00 ± 11.14 68.67 ± 14.98 88.67 ± 30.92 4 weeks 69.67 ± 22.55 65 ± 7.81 74.67 ± 19.14

As shown in the results shown in Tables 4 and 5, the softening water containing the extract of Sotoja reduced acne worsening and inhibited sebum secretion in each part of the face even in those suffering from sebum secretion. .

As can be seen in the examples and experiments according to the present invention, the water extracts provided by the present invention not only have an excellent 5α-reductase inhibitory effect but also reduce acne and reduce sebum secretion when used as a cosmetic composition in humans. The effect is excellent. Therefore, the water extract according to the present invention has an acne laxative effect or sebum secretion inhibitory effect at a concentration that does not affect the safety of the product, so it can be used as an effective concentration in sebum inhibitory cosmetics or acne laxative cosmetics, and general dosage forms of medicines for skin application It can be used as an active ingredient in.

Claims (3)

5α-reductase inhibitor, characterized in that it contains a water extract extracted with methanol or ethanol. The 5α-reductase inhibitor of claim 1, which reduces side effects due to dihydrotestosterone. A sebum inhibiting or acne treatment cosmetic or skin coating drug, comprising the extract from methanol or ethanol according to claim 1.