JP4216173B2 - Composition effective in preventing and improving pigmentation caused by stress - Google Patents

Description

The present invention relates to an inhibitor containing an extract of elcampuri and capable of specifically antagonizing the tyrosinase activity of melanocytes promoted via adrenocorticotropic hormone (ACTH).

  Conventionally, for the purpose of preventing and improving skin pigmentation caused by ultraviolet rays, substances that inhibit the activity of tyrosinase, which catalyzes the production of melanin, and substances that have the effect of reducing the produced melanin pigment have been studied. Has been applied to drugs. In addition, for the purpose of reducing the amount of ultraviolet irradiation to the skin, an external preparation for skin containing an ultraviolet absorbent such as a paraaminobenzoic acid compound or a benzophenone derivative or a powder having an ultraviolet shielding effect such as titanium oxide or zinc oxide is applied. I came. Furthermore, it became clear that cytokines such as α-melanocyte stimulating hormone (α-MSH), endothelin, stem cell growth factor (SCF), histamine, and active oxygen are involved in the activation of melanocytes by ultraviolet rays. Prevention and improvement of pigmentation symptoms by controlling these factors have been studied.

However, in recent years, it has been clarified that pigmentation symptoms are caused not only by ultraviolet rays but also by stress (see Non-Patent Document 1). In general, aggravation of a stain due to stress is different from a stain caused by ultraviolet rays, and is different from that caused by anterior pituitary gland (Adrenocorticotropic hormone).
(Hormone: ACTH) is considered to be involved. ACTH is a hormone that promotes the production of steroid hormones in the adrenal cortex, but it is a precursor of α-MSH and also has the property of promoting the production of melanin, and its secretion increases due to stress and the like. There is a feature to do.
Hironori Kaminaga, Tatsuro Shinomiya, Journal of the Japanese Society of Skin, 107 (5): 615-622, 1997.

ACTH in blood increased by stress is considered to be activated by binding to melanocortin receptor-1 on the cell membrane surface of melanocytes scattered in the epidermis. In activated melanocytes, tyrosinase synthesis is promoted and melanin production is increased, which is thought to lead to pigmentation. In addition, addison's disease with systemic pigmentation is also caused by excessively secreted ACTH in the blood due to decreased adrenal function, and endogenous ACTH may be involved in skin pigmentation. It is done. As described above, the activation of melanocytes induced by stress is mainly the activation of melanocytes via ACTH, and conventional skin external preparations intended to prevent and improve skin pigmentation due to ultraviolet rays are sufficient. In some cases, the effect cannot be expected. Therefore, it is considered important to prevent activation of melanocytes by ACTH in order to prevent and improve pigmentation symptoms due to stress. Corticostatin has been reported as a component that prevents activation of melanocytes by ACTH (see Non-Patent Document 2), but there are problems such as being unstable and being a biological component and difficult to purify.
Inoue K. , J. et al. Invest. Dermatol. 121: 165-171, 2003.

The Elcampuri used in the present invention is a plant belonging to the Gentianaceae family, and the scientific name is Gentiana prostrata (Gentiana).
There is also a theory that it is called alborose). It grows naturally in the Amazon region of South America and is generally called Hercampuri in Peru. It is a medicinal herb with a strong bitter taste, and the whole herb has an effect of promoting bile excretion, and is used as a thinning drug or a therapeutic drug for hepatitis.

In addition, about the extract of elcampuri, having tyrosinase inhibitory action and suppressing melanin production | generation are disclosed (refer patent document 1, 2). However, the extract of Elcampuri has an inhibitory action on ACTH, and it has not been reported so far to suppress melanin production activated by stress through the action.
Japanese Patent Laid-Open No. 08-175963 JP 2001-106619 A

An object of the present invention is to provide a tyrosinase activity inhibitor that suppresses the tyrosinase activity of melanocytes promoted through ACTH and , as a result, effectively prevents and improves pigmentation caused by stress. To do.

Under these circumstances, as a result of intensive studies, the present inventors have found that the extract of Elcampuri has an excellent inhibitory effect on tyrosinase activity of melanocytes promoted by ACTH, and completed the present invention. It came to do.

An inhibitor of melanocyte tyrosinase activity promoted by adrenocorticotropic hormone containing an extract of elcampuri of the present invention specifically antagonizes ACTH produced by stress and promotes melanocyte tyrosinase via ACTH Inhibiting the activity is effective in preventing and improving pigmentation symptoms caused by stress.

  The extract of Elcampuri used in the present invention is obtained by extracting from a part of whole plant such as leaves, stems, flowers, seeds, fruits, rhizomes, roots, etc. or whole plants. Preferably, one obtained by extraction from one of leaves or stems or a mixture of both is preferable.

  In addition, the extract of elcampri used in the present invention is obtained by immersing or heating elcampuri with an extraction solvent, followed by filtration and, if necessary, concentration. Examples of the extraction solvent include water, lower monohydric alcohols (methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, etc.), liquid polyhydric alcohols (1,3-butylene glycol, propylene). Glycol, etc.), hydrocarbons (hexane, pentane, etc.), ketones (acetone, methyl ethyl ketone, etc.), ethers (ethyl ether, tetrahydrofuran, propyl ether, etc.), acetonitrile and the like. These solvents may be used alone or in combination of two or more. Preferably, one or two or more of water or a water-soluble solvent (solvent that can be mixed with water in an arbitrary ratio. For example, ethanol, 1,3-butylene glycol, propylene glycol, etc.) may be used. . The extract may be used as it is, or a part or all of the solvent may be distilled off.

  The Elcampri extract used in the present invention can be used in any of cosmetics, quasi-drugs, and pharmaceuticals. Examples of the dosage form include lotions, creams, emulsions, gels, aerosols, and poultices. , Essence, pack, cleaning agent, bath preparation, foundation, dusting powder, lipstick, ointment, shampoo, rinse, treatment, tonic and the like.

The compounding amount of the extract of Elcampri used in the present invention is preferably 0.0001 to 0.001% by weight as a dry product. The addition method may be added in advance or during the production, and may be appropriately selected in consideration of workability.

  Next, in order to describe the present invention in detail, examples of production of the extract used in the present invention, formulation examples and experimental examples will be given as examples, but the present invention is not limited thereto. The compounding amount shown in the examples indicates% by weight.

Production Example 1 Elcampri hot water extract 400 mL of water was added to 20 g of Elcampuri stems and leaves, extracted at 95-100 ° C. for 2 hours, the insoluble matter was filtered, the filtrate was concentrated, dried and dried. 4.3 g of water extract was obtained.

Production Example 2 Elcampriethanol extract 900 ml of 80% ethanol was added to 100 g of Elcampuri leaves, extracted at room temperature for 7 days, insoluble matter was filtered, and the filtrate was concentrated to dryness to obtain 33 g of Elcampriethanol extract. Obtained.

Production Example 3 Elcamppri 1,3-butylene glycol extract 1000 ml of a mixture of 1,3-butyleneglycol and water (1: 1) was added to 30 g of a dry product of whole Elcamppri, and extracted at room temperature for 10 days. Insoluble matter was filtered to obtain 950 g of Elcamppri 1,3-butylene glycol extract.

  Next, experimental examples will be given to explain the effects of the present invention in detail.

Experimental Example 1 Inhibition test on activation of melanocytes by ACTH The effect of inhibiting the activation of melanocytes by ACTH was measured under the following conditions. That is, B-16 mouse melanoma cells were used as melanocytes, and activation of melanocytes promoted by addition of ACTH was measured using the amount of tyrosinase synthesized in melanocytes as an index. A sample was added to this reaction system, and the production inhibition rate of the amount of tyrosinase synthesis was calculated to obtain the effect of inhibiting the activation of melanocytes by ACTH.

  B-16 mouse melanoma cells were seeded in 96-well micro-plates at 3.2 × 10 4 per well, and cultured in Eagle's MEM culture medium containing 1% FCS at 37 ° C. under 5% CO 2 for 2 days. Next, the medium was changed to Eagle's MEM culture medium containing 1% FCS supplemented with 1 or 10 μg / mL of Elcampuri extract and 40 nM ACTH, and further cultured for 2 days. Thereafter, the activity of tyrosinase produced in the cells was colorimetrically determined. That is, after removing the medium, the plate was washed with PBS, 50 μL of 1% Triton X-100 PBS solution was added, and the cells were lysed by stirring for 30 minutes, and the tyrosinase activity and protein amount of the lysate were measured. Tyrosinase activity was determined by adding 100 μL of 0.25% L-DOPA solution to 25 μL of 1% Triton X-100 PBS cell lysate, incubating at 37 ° C. for 3 hours, measuring the absorbance at 450 nm, and separately measuring the protein. Corrected by amount. About the obtained tyrosinase production amount, the activation suppression rate of the melanocyte was computed from the decreasing rate of the tyrosinase production amount at the time of the sample addition with respect to control.

The test results are shown in Table 1. As a result, an excellent inhibitory effect on the activation of melanocytes by ACTH was observed in the Elcampuri extract. Moreover, as a result of conducting the same test without adding ACTH, the Elcampuri extract did not affect the amount of tyrosinase synthesized in melanocytes at sample concentrations of 1 and 10 μg / mL. Therefore, the effect of the ercampuri extract directly on the melanocytes without ACTH is very small, and it can be said that this test shows the inhibitory effect of the ercampli extract on the activation of melanocytes by ACTH.

  As an application example of the present invention, it can be used for any of cosmetics, quasi drugs and pharmaceuticals. As the dosage form, for example, lotion, cream, milky lotion and the like can be mentioned, and when applied externally, the effect of preventing and improving pigmentation caused by stress is expected.

Claims (1)

An inhibitor of tyrosinase activity of melanocytes promoted via adrenocorticotropic hormone, characterized by containing 0.0001 to 0.001% by weight of a hot water extract of Elcampuri or 80% ethanol extract as a dried product.