JP2016216365A - Pharmaceutical composition for repairing and activating skin and method for producing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an agent that repairs and activates the skin damaged due to ultraviolet rays and the like and has few adverse effects.SOLUTION: A pharmaceutical composition is extract from lotus young leaves, mature leaves, stems, roots or seeds, and is used to activate the skin by the redifferentiation to a normal cell and the redifferentiation of a damaged cell to the normal cell.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a composition for repairing and activating damaged skin and a method for producing the same, and more particularly, to a skin repair activator based on a substance contained in a plant and the production thereof. It is about the method.

Currently, methods for activating human skin cells include 1) physical methods such as skin massage and thermotherapy, 2) chemical methods such as plant extracts and artificial drugs applied to the skin, 3 ) Oral administration of vitamins and the like is common.

In recent years, a plant-derived natural component has attracted attention as a substance having a high skin activity and few side effects (Non-patent Document 1). For example, henna and prune have been found to have anti-inflammatory and anti-allergic effects (Patent Documents 1 and 2).

Mulberry (mulberry) leaves have been pointed out to have a skin whitening effect and ultraviolet blocking effect, and their use is expected (Patent Documents 2 and 3).

Lotus has a lot of catechins, which are polyphenols, and by taking a certain amount of plant-derived flavonoids, it has been pointed out that it has antioxidant, anti-inflammatory and antibacterial effects (Patent Document 4).

It is said that lotus also has an antiallergic effect by suppressing histamine release from mast cells (Patent Document 5).

Living herbs and herbal medicines Mizuo Mizuno, Mizuo Ota New Japan Law Publication KK. August 2014

JP 2001-335496 A

JP 2003-267882 A

JP 2005-298511

JP 2001-158742 A

JP2006-151811

However, in Patent Documents 4 and 5, there is no scientific evidence such as cell proliferation effect of lotus extract on skin epithelial cells or improvement of resistance to ultraviolet rays, and effective concentration and administration on skin epithelial cells. The quantity has not been optimized.

Furthermore, the development of a drug that repairs and activates skin damaged by ultraviolet rays or the like has not progressed sufficiently.

The present invention has been made in view of the above problems, and its purpose is to establish a method for extracting an active ingredient from a plant and use the extract to repair damaged skin and to activate it. It is to realize a drug with less side effects.

In order to solve the above-mentioned problems, the pharmaceutical composition of the present invention is a pharmaceutical composition for activating skin containing an extract extracted from lotus.

In the pharmaceutical composition of the present invention, the extract is preferably extracted from the young leaves, mature leaves, stems, roots or seeds of the lotus.

In the pharmaceutical composition of the present invention, the extraction liquid is preferably water-soluble.

The pharmaceutical composition of the present invention is preferably in the form of skin application.

In order to solve the above-mentioned problem, the method for producing a medicament of the present invention is a method for producing a medicament for activating skin, including a step of obtaining an extract extracted from a lotus using a polar solvent.

In order to perform extraction effectively, the inside of the extraction furnace is negatively ionized and also includes a low pressure state.

In the pharmaceutical production method of the present invention, the extract is preferably extracted from the young leaves, mature leaves, stems, roots or seeds of the lotus.

In the pharmaceutical production method of the present invention, the polar solvent is preferably water.

In order to solve the above-mentioned problems, the pharmaceutical composition of the present invention is characterized by containing an extract extracted from the young leaves, mature leaves, stems, roots or seeds of the lotus.

By using the present invention, the skin can be activated by redifferentiation of normal cells and damaged cells into normal cells without causing side effects, and the amount of ultraviolet irradiation is increasing. It is thought that it will greatly help with countermeasures.

(1. Pharmaceutical composition and method for producing the same) The pharmaceutical composition of the present embodiment contains an extract extracted from lotus, and redifferentiates normal cells and damaged cells into normal cells. Is used to activate the skin. Below, the pharmaceutical composition of this Embodiment and its manufacturing method are demonstrated.

The lotus used in the present invention is not particularly limited, and a known lotus can be used as appropriate. The lotus may be a wild type, but an artificially cultured lotus may be used. This time, Iwakuni lotus was used.

Further, the lotus site for extracting the extract is not particularly limited. For example, it is preferable to use young lotus leaves, mature leaves, stems, roots or seeds as the lotus site.

The polar solvent used for extraction is not particularly limited. For example, water, an aqueous alcohol solution (for example, methanol, ethanol, propanol, etc.), or a mixed solution thereof is used. In addition, it is desirable that the best extraction method is to extract the inside of the extraction furnace with an extract extraction structure (apparatus) under a negative ion atmosphere at a low pressure. Therefore, the extract is extracted from 100% plants. From the viewpoint of effectively extracting a target substance and easily preparing a pharmaceutical composition, it is also considered to use water among the polar solvents.

Although the temperature of the said polar solvent is not specifically limited, It is although it does not specifically limit as said extraction furnace surface temperature, For example, it is preferable that it is 50 to 100 degreeC, It is more preferable that it is 70 to 100 degreeC, 90 degreeC Most preferably, it is -100 degreeC. According to the said structure, the extraction effect of the target substance can be raised, without reducing a medicinal effect. In view of the various matters described above, it is most preferable to use hot water (for example, boiling water) as the polar solvent.

The specific operation for extracting the extract from the lotus is not particularly limited, and may be appropriately performed based on a known method. For example, extraction can be performed using an ultrasonic irradiator, a stirrer, a heater, a pressurizer, a vacuum device, or the like. For example, when water is used as the solvent, a target substance may be extracted by boiling lotus with boiling water. In addition, although it does not specifically limit as boiling time, For example, it is preferable that it is 10 minutes-30 minutes, It is more preferable that it is 15 minutes-25 minutes, It is most preferable that it is 20 minutes. According to the said structure, the said substance can be extracted effectively, without applying useless heat to the target substance. During the extraction operation, it is possible to perform ultrasonic treatment on the extract or to stir the extract.

The pharmaceutical composition of the present embodiment can be used to activate skin by redifferentiation of normal cells and damaged cells into normal cells.

The pharmaceutical composition of the present embodiment may contain a pharmaceutically acceptable carrier in addition to the lotus extract. The pharmaceutically acceptable carrier used in the pharmaceutical composition can be selected depending on the dosage form and dosage form of the pharmaceutical composition.

As used herein, as a pharmaceutically acceptable carrier, various organic or inorganic carrier materials that can be used as a formulation material are used. It can be incorporated into a pharmaceutical composition as a bulking agent, binder, or disintegrant, or as a solvent, solubilizer, suspension, isotonic agent, buffer, or soothing agent in a liquid preparation.

Although it does not specifically limit as said excipient | filler, For example, lactose, sucrose, D-mannitol, xylitol, sorbitol, erythritol, starch, crystalline cellulose etc. can be used.

Although it does not specifically limit as said lubricant agent, For example, a magnesium stearate, a calcium stearate, a talc, colloidal silica, etc. can be used.

The binder is not particularly limited, and for example, pregelatinized starch, methylcellulose, crystalline cellulose, sucrose, D-mannitol, trehalose, dextrin, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone and the like can be used.

Although it does not specifically limit as said disintegrating agent, For example, starch, carboxymethylcellulose, low substituted hydroxypropylcellulose, carboxymethylcellulose calcium, croscarmellose sodium, carboxymethyl starch sodium, etc. can be used.

Although it does not specifically limit as said solvent, For example, water for injection, alcohol, propylene glycol, macrogol, sesame oil, corn oil, tricaprylin etc. can be used.

The solubilizing agent is not particularly limited, and for example, polyethylene glycol, propylene glycol, D-mannitol, trehalose, benzyl benzoate, ethanol, trisaminomethane, cholesterol, triethanolamine, sodium carbonate, sodium citrate and the like are used. obtain.

The suspending agent is not particularly limited. For example, a surfactant such as stearyltriethanolamine, sodium lauryl sulfate, laurylaminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, glyceryl monostearate, or polyvinyl Hydrophilic polymers such as alcohol, polyvinylpyrrolidone, sodium carboxymethylcellulose, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose and the like can be used.

Although it does not specifically limit as an isotonizing agent, For example, sodium chloride, glycerol, D-mannitol etc. can be used.

Although it does not specifically limit as a buffering agent, For example, buffer solutions, such as a phosphate, acetate, carbonate, a citrate, etc. can be used.

Although it does not specifically limit as a soothing agent, For example, benzyl alcohol etc. can be used.

Although it does not specifically limit as antiseptic | preservative, For example, paraoxybenzoic acid ester, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, sorbic acid etc. can be used.

Although it does not specifically limit as an antioxidant, For example, a sulfite, ascorbic acid, etc. can be used.

The pharmaceutical composition of the present embodiment can be produced by a known method in the pharmaceutical field.

The lotus extract content in the pharmaceutical composition of the present embodiment can be set in consideration of the administration form, administration method and the like. For example, there is no particular limitation as long as the lotus extract can be administered within the dosage range described below when the pharmaceutical composition is used.

When the pharmaceutical composition of the present embodiment is in the form of an oral preparation, for example, starch, lactose, sucrose, mannitol, carboxymethylcellulose, corn starch, inorganic salts and the like can be used as a pharmaceutical carrier. Moreover, when preparing an oral preparation, you may mix | blend a binder, a disintegrating agent, surfactant, a lubricant, a fluidity promoter, a corrigent, a coloring agent, a fragrance | flavor, etc. further.

When the pharmaceutical composition of the present embodiment takes the form of a parenteral agent, according to a method known in the art, distilled water for injection, physiological saline, aqueous glucose solution, vegetable oil for injection, sesame oil, By dissolving or suspending the active ingredient of the present invention in peanut oil, soybean oil, corn oil, propylene glycol, polyethylene glycol, etc., and adding a disinfectant, a stabilizer, an isotonic agent, a soothing agent, etc., if desired. The pharmaceutical composition of the present embodiment can be prepared.

The pharmaceutical composition of the present embodiment can be administered by an appropriate administration route according to the preparation form. There is no particular limitation on the administration method, and it can be administered internally, externally, or by injection. If the pharmaceutical composition of the present embodiment has a form as an injection, it can be easily administered, for example, intravenously, intramuscularly, subcutaneously, or intradermally. Therefore, the pharmaceutical composition of the present embodiment may be a fluid, a solid, or an aerosol.

The dosage of the pharmaceutical composition of the present embodiment can be appropriately set depending on the form, administration method, purpose of use, and age, weight, and symptom of the patient to whom the pharmaceutical is administered. In general, the dose of the active ingredient contained in the preparation is preferably 0.1 to 2000 mg / kg per day for an adult. Of course, the dose may vary depending on various conditions, and an amount smaller than the above dose may be sufficient, or an amount larger than the above dose may be required.

Administration may be performed once within a day within a desired dose range, or may be performed in several divided portions. Further, the pharmaceutical composition of the present embodiment can be applied to the skin as it is, or added to any food or drink and taken on a daily basis.

Pharmaceutically acceptable excipients that may be used in the pharmaceutical composition of the present embodiment generally constitute 5% to 99.9%, preferably 25% to 80% of the weight of the composition, and In the absence of other adjuvants, it may constitute the remainder of the composition. Preferably, at least 80% by weight of the excipient is water. Preferably, water constitutes at least 50%, most preferably 60-80% by weight of the pharmaceutical composition.

[Example 1: Effect of lotus extract on human epithelial cells]
The lotus was extracted in a low-pressure atmosphere with a negative ion atmosphere inside the extraction furnace. The obtained extract was about 50 percent by weight of the lotus, and this extract was passed through a sterilized filter to obtain a sterilized extract.

Next, the cells to be examined for skin activation of the lotus extract will be described.

The cells used were NHSF46 Japanese skin-derived normal diploid fibroblast cell lines provided by RIKEN Cell Bank.

Subsequently, NHSF46 Japanese skin-derived normal diploid fibroblast cell line was cultured in MEMα + 10% FBS medium at a CO ₂ concentration of 5% at 37 ° C. for 3 days.

The cultured cells were trypsinized, and the collected cells were added to MEMα + 10% FBS medium to adjust the number of cells to 5 × 10 ⁴ cells / ml. 100 μl of this cell-containing medium was added to each well of two 96-well plates and cultured at 37 ° C. for 1 day. One of the two 96-well plates was irradiated with a distance of 60 cm from a 15 W ultraviolet lamp for 30 minutes to prepare two groups, an ultraviolet irradiation group and an ultraviolet non-irradiation group. In each of the two groups, the lotus extract was diluted at the time of use and replaced with medium adjusted to 0% (no lotus solution added) and lotus concentrations of 5, 10, 15, and 20%, and the carbonic acid at 37 ° C was replaced. After culturing in a gas incubator for one day, cell morphological observation and cell proliferation rate were determined.

The cell growth rate was calculated by measuring the number of living cells using the Cell Counting Kit of Dojindo Laboratories. The experiment was performed three times and statistical processing was performed.

As a result, FIG. 1 and FIG. 2 are morphologically observed. Morphologically, the spindle-shaped morphology was slightly swollen by UV irradiation, but showed little change. FIG. 3 shows the result of an epithelial cell proliferation test using a lotus solution of epithelial cells affected by this degree of ultraviolet light.

Non-irradiated cells were found to have an increased cell growth rate at 5% in the lotus extract-added medium. However, if the concentration exceeds 5%, the proliferation rate decreases as the concentration increases, and it has been shown that excessive administration reduces cell proliferation ability. In UV irradiated cells, the growth rate of cells without lotus extract is reduced to 40% or less, but when 5% of lotus extract is added, the growth rate is improved to about 60%, but the lotus extract concentration is 10%. Above%, the growth rate is constant. It has been found that the addition of 5% concentration of lotus extract has a great effect on restoring the growth ability of UV-irradiated cells and improving the growth ability of non-irradiated cells.

[Example 2: Effect of lotus extract on type 1 collagen mRNA and estrogen α receptor mRNA expression in epithelial cells]
Experiments were performed using the lotus extract used in Example 1 and the NHSF46 Japanese skin-derived normal diploid fibroblast cell line.

⁴ ml of MEMα + 10% FBS medium adjusted to 5 × 10 ⁴ cells / ml was seeded in a 6 cm dish and cultured at 37 ° C. for 1 day. The lotus extract is diluted at the time of use and replaced with a medium adjusted to 0% (no lotus solution added) and lotus concentration of 5, 10, 15, 20%, and cultured for one day in a 37 ° C carbon dioxide incubator. did.

The collected RNA was examined for the expression state of type I collagen mRNA and estrogen α receptor mRNA by RT-PCR. The results are shown in FIGS.

Type I collagen is a fibrous collagen that is contained in large amounts in bones, making the bones elastic, and also in the dermis of the skin, resulting in strength of the skin. Increase collagen production. Moreover, the collagen production of skin cells is greatly influenced by female hormones (estrogens), and the increase in estrogen α receptor is involved in the increase in type 1 collagen production. Thus, an increase in estrogen alpha receptor mRNA increases type 1 collagen production, leading to skin activation and elasticity.

As shown in FIG. 4, the expression of type I collagen mRNA increases about 2.5 times in the medium containing lotus supplemented solution 5% and 10% compared to the non-added solution, and decreases when added at a concentration higher than that. Yes.

Moreover, as shown in FIG. 5, estrogen α receptor mRNA is expressed 3.5 times as much in the 5% concentration medium with lotus added solution and 2.5 times when 10% concentration is added as compared with no addition, but decreases when the concentration is higher than that. It was shown that.

From the above results, epithelial cells increase the expression of type 1 collagen mRNA and estrogen α receptor mRNA by adding 5% concentration of lotus extract, and increase the production of collagen in the skin, resulting in increased skin activity and elasticity. It has been shown to improve.

From Example 1 and Example 2, it was found that the extract extracted from lotus promotes the growth of normal human epithelial cells, and further helps to recover epithelial cells damaged by ultraviolet rays. It was found to be 5%.

Note that the present invention is not limited to the configurations described above, and various modifications are possible within the scope of the claims, and technical means disclosed in different embodiments and examples respectively. Embodiments and examples obtained by appropriately combining them are also included in the technical scope of the present invention.

By using the present invention, normal skin cells can be activated and the elasticity of the skin can be increased, and at the same time, damaged skin can be repaired and activated, and a drug with less side effects can be provided, thus greatly contributing to the development of the pharmaceutical field. can do.

It is a figure of the cell when ultraviolet rays are irradiated to the cell derived from MKN45 human epithelium. It is a figure of the cell of which MKN45 human epithelial origin cell is not irradiated with ultraviolet rays. It is a figure which shows the proliferation effect with respect to the MKN45 human epithelial origin cell by a lotus extract. It is a graph which shows collagen 1 mRNA expression with respect to the MKN45 human epithelial origin cell by a lotus extract. It is a graph which shows the estrogen receptor alpha mRNA expression with respect to the MKN45 human epithelial origin cell by a lotus extract.

Claims (6)

A pharmaceutical composition for activating skin by redifferentiation of normal cells and damaged cells into normal cells, which is an extract extracted from a lotus. The pharmaceutical composition according to claim 1, wherein the extract is extracted from young leaves, mature leaves, stems, roots or seeds of the lotus. The pharmaceutical composition according to claim 1 or 2, wherein the extract is a liquid.   The pharmaceutical composition according to claim 1, 2 or 3, wherein the pharmaceutical composition is in the form of skin application. A method for producing a medicament, wherein the extract activates skin by redifferentiation of normal cells and damaged cells into normal cells. The method for producing a medicine according to claim 5, wherein the extract is extracted from young leaves, mature leaves, stems, roots or seeds of the lotus.

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