KR20000024876A - Dermal composition for curing wound containing collagen synthesis promoter - Google Patents

Abstract

PURPOSE: A dermal composition for curing wound containing more than one of extract from magnolia floss, Angelicae Dahuricae Radix, Caryophylli floss as a collagen synthesis promoter is provided, which shows strong collagen synthesis promoting effect and excellent wound curing effect when applied on skin, and has prominent safety without side effect. CONSTITUTION: A preparing process of dried extract from magnolia floss, Angelicae Dahuricae Radix, Caryophylli floss comprises of: crushing the herb medicine; adding 5-20volume of water, primary alcohol, ethylacetate, acetone and chloroform; heating and extracting at 50-100°C for 1-5hrs; filtering extract; extracting more than one time using the same method; adding the extracts from each stage together; concentrating under pressure reducing state; freeze-drying or spray-drying. The extract can be used in preparing ointment or cosmetic material, of which content in the ointment or cosmetic material is 0.0001-10.0wt.%, desirably 0.0005-10wt.% based on dried weight.

Description

Skin composition for wound healing comprising collagen synthesis promoter

The present invention relates to a skin composition for wound healing, and more particularly, to a skin medicine and a cosmetic having an excellent wound healing effect including a shinto extract, white paper extract and / or cloves extract as a collagen synthesis promoter.

Collagen, a major component of the extracellular matrix, is a major matrix protein produced by skin fibroblasts and is an important protein in extracellular epilepsy and accounting for about 30% of the total weight of biological proteins. Have. Collagen forms most of the organic material of the skin, tendons, bones and teeth, especially in bones and skin (dermis). Most other sieve structures exist as fibrous inclusions. Collagen is a relatively weak immunogen, partly due to the shielding of latent antigenic determinants by the helix structure of collagen, which also makes collagen resistant to proteolysis. The main functions of collagen are known as mechanical firmness of skin, resistance of connective tissue and binding of tissue, support of cell adhesion, induction of cell division and differentiation (in organic growth or wound healing) (Van der Rest et al. 1990). As such, collagen, which has important functions in vivo, is biosynthesized in fibroblasts, and is reduced by aging and photoaging due to UV irradiation. In general, the amount of collagen present in the skin in normal people is 65% during 20 to 80 years. It is known to decrease the thickness to make the skin thinner (Shuster S., British Journal of Determatology, 93, 639 (1975)). The metabolic turnover rate of collagen decreases with age, causing hardening of the extracellular matrix. Collagen also plays an important role in wound healing, promoting the synthesis of collagen in the damaged epithelium so that wounds can be quickly recovered without scarring.

As a result of extensive research on collagen, important functions of collagen have been revealed in the skin, and if collagen metabolism is activated by promoting collagen synthesis in vivo, dermal matrix components are increased, resulting in wound healing, elasticity enhancement, skin strengthening, etc. It turns out that you can expect.

In order to obtain such effects, conventionally, products using collagen have been released in order to take advantage of collagen's skin moisturizing effect.However, these cosmetics, such as applying collagen to the surface of the skin, are difficult to absorb the percutaneous polymer of collagen. I couldn't expect it to be an essential skin function improvement.

In order to solve these problems, interest in collagen synthesis promoting substances has been increased, and conventional collagen synthesis promoting substances include vitamin C and its derivatives, vitamin A and its derivatives, insulin, growth hormone, estrogen, and beta-type tumor growth factors ( Tumor growth factor-beta, Dansam root extract (JP 2-290805), animal placental protein (JP 8-231370), mammalian whey (JP 3-20206), chlorella extract (JP 9-40523, JP 10 -36283, fibroblast proliferation promoting effect) and the like are known.

However, conventional collagen synthesis promoting substances are limited in use or have a weak effect due to problems such as biosafety, and thus cannot substantially expect an improvement in skin function by promoting collagen synthesis of the skin. Therefore, there is an urgent need for development of new collagen synthesis promoters which are safe for living bodies and are more effective than conventional collagen synthesis promoters.

The present invention is to solve the problems of the prior art as described above, and an object of the present invention is to provide a composition for skin that is safe for living beings and has a high effect of promoting collagen synthesis.

In order to achieve the above object, the present invention provides a skin composition for wound healing, comprising one or more extracts selected from the group consisting of shinyi extract, white paper extract and clove extract as a collagen synthesis promoter.

Hereinafter, the present invention will be described in detail.

The present inventors have conducted research to develop an excellent collagen synthesis promoter for natural native plants and animals, and found that the god extract, white paper extract and cloves extract have a very strong collagen synthesis promoting effect and completed the present invention. .

The skin composition of the present invention comprises one or more extracts selected from the group consisting of Sinyi extract, white paper extract and clove extract as a collagen synthesis promoter, and these extracts are available as medicinal herbs as a primary extract. The mixture was first extracted with water or an organic solvent such as anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms, ethyl acetate or acetone, and then concentrated under reduced pressure, followed by freeze drying or spray drying.

Singi, white paper and clove, which are herbal medicines used in the present invention, have the following characteristics.

The deciduous shrub is a deciduous shrub, Latin name is Flos Magnoliae, cultivated in gardens in southern China, and wild. It belongs to Magnoliaceae, and it is picked up and dried in the sun before spring buds open. The plant of origin is Magnolia denudata. Besides, the Korean herbal medicine standard contains magnolia kobus, Magnolia salicifolia Maxim.The Chinese Pharmacopoeia defines Magnolia biondii, Magnolia sprengeri, Magnolia liliflora, Magnolia faragesii. Shini is traditionally known to relieve soreness in the wind, to remove freckles on the face, to relieve noses and runny noses, to brighten the eyes, and to produce beards and hairs (herb wood), The effect of is distinguished from the collagen synthesis promoting effect of the present invention. In addition, research has been reported on cosmetic compositions (Japanese Patent Laid-Open Publication No. 5-320034), skin disease treatment agents (Japanese Patent Laid-Open Publication No. 5-246874), deodorant (Japanese Patent Laid-Open Publication No. 5-212099), etc., which have a skin whitening effect and an anti-inflammatory effect in relation to shinyi. However, it is completely distinct from the present invention. Existing god-identification studies have reported substances such as Magnolol, Syringin, Honokiol, Eugenol, and Thyramine.

The white paper originates from the copper stalk, Umbelliferae, whose scientific name is Angelica dahurica and Latin name is Radix Angelicae Dahuricae. Perennial herb, growing in grassy wetlands, shrubbery or riverside. Collect roots in autumn and dry them in the sun. Existing ingredient identification studies show that Kumarins, Byak-angelicin, Oscine, Xangthotoxin, (-)-Nokadekenetin, etc. The substance is known. White paper is traditionally known to cure head soreness due to heat or wind. Herbal medicine, pain relief, hemostatics, used as a sedative. In studies related to white paper, the treatment of dermatitis by promoting sebum secretion (Japanese Patent Laid-Open No. 9-176030), hair growth inhibitor with excellent human stability (Japanese Patent Laid-Open No. 10-139639), and bathing agents with increased warming effect (Japanese Patent Laid-Open No. 06-206814) And the like, but are completely distinguished from the collagen synthesis promoting effect of the present invention.

Cloves are Mytaceae, scientific name is Eugenia aromaticum or Eugenia caryophyllata, Syzygium aromaticum, and Latin name is Flos Caryophylli. When buds are safflower color, they are collected and dried in the sun. Cloves from the herbaceous wood are known to have hair growth effects, and they are used for abdominal pain and earth and sand. Clove ethanol extract has a contraction of the extracted uterus, local anesthesia and mild analgesic action is used for tooth decay. There is also a dipharytic and antiviral action (Yamahara, J., et al: J., pharm. Dyn., 6.281, 1998.3). Prior studies have shown that whitening action (Japanese Patent Laid-Open Publications 5-155735, 6-16530, 6-24955), antioxidants (Japanese Laid-Open Patent Publication 5-320037), and antiallergic agents (Japanese Patent Laid-Open Publications 7-215846, 10-36276 ), Antibacterial action (Japanese Patent Laid-Open No. 9-255517, 10-45562) and the like are known, but this is completely distinguished from the collagen synthesis promoting effect of the present invention. Conventional clotting component identification studies have reported substances such as eugenol, chavicol, higenamine, vanillin and the like.

The preparation method of the herbal medicine extract is as follows. First, a commercially available medicinal herb purchases white paper and cloves, and finely grinds them, and 5-20 volumes of water, 1-4 anhydrous or hydrous lower alcohols, ethyl acetate, acetone, chloroform or theirs based on the dry weight of the ground powder. The mixture was added and extracted with warming at 50-100 ° C. for 1-5 hours in an extractor with reflux condenser. The residue was filtered from the extract with a filter cloth and then extracted one or more times in the same manner. The extracts obtained in each step were concentrated under reduced pressure, and freeze-dried or spray-dried to obtain a dry extract.

Sinyi extract, white paper extract and clove extract produced by this method showed a very powerful collagen biosynthesis promoting effect in fibroblasts, and when applied to the wound site of the rat showed a very good wound healing effect.

In addition, it is possible to prepare an external skin ointment or cosmetic including such god extract, white paper extract and clove extract, the content of the god extract, white paper extract and / or clove extract included in the manufacture of external skin ointment or cosmetics in dry weight. 0.0001-10.0% by weight, preferably 0.0005-10% by weight. If the content of the extract is less than 0.0001% by weight, no obvious effect could be expected. If the content of the extract was more than 10% by weight, the effect of the extract was not increased.

Cosmetics such as skin external ointments or lotions containing these god extracts, white paper extracts, and / or clove extracts have a very good wound healing effect when applied to the wound site of a rat incision and have no side effects. The extract, white paper extract and clove extract were found to be a safe and effective collagen biosynthesis promoter.

EXAMPLE

The following presents preferred examples and comparative examples to aid in understanding the invention. However, the following examples are merely provided to more easily understand the present invention, and the present invention is not limited to the following examples.

Example 1-24

300 g of a commercially available medicinal herb obtained by pulverizing was added to 3 L of a solvent, extracted by boiling in a reflux extractor equipped with a cooling condenser for 3 hours, filtered through a 300 mesh filter cloth, and the residue was extracted once more in the same manner. The extracts obtained in each step were mixed and filtered with Whatman No. 2 filter paper at room temperature to remove insoluble matters, and then concentrated under reduced pressure to 60 ° C. in a distillation apparatus equipped with a cooling condenser and freeze-dried to obtain a dry extract. The dry weights of the herbs, solvents and dry extracts obtained in each case are shown in Table 1.

Example Herbal medicine menstruum Dry weight (g) One God 80% methanol 32 2 Purified water 37 3 85% ethanol 32 4 70% acetone 29 5 50% ethanol 34 6 60% methanol 25 7 Saturated normal butanol 27 8 chloroform 25 9 blank 80% methanol 50 10 Purified water 53 11 85% ethanol 55 12 70% acetone 60 13 50% ethanol 57 14 60% methanol 51 15 Saturated normal butanol 54 16 chloroform 52 17 cloves 80% methanol 35 18 Purified water 32 19 85% ethanol 34 20 70% acetone 34 21 50% ethanol 36 22 60% methanol 35 23 Saturated normal butanol 37 24 chloroform 32

Experimental Example 1 Collagen Biosynthesis Effect at the Cell Level of Sinyi Extract, White Paper Extract and Clove Extract

Each extract obtained in Example 1-24 was added to the fibroblast culture medium derived from human, and the collagen biosynthesis promoting effect at the cellular level was tested. In this case, the extracts obtained in Examples 1-24 were prepared to have a final concentration of 0.001w / v% to measure collagen biosynthesis promoting effect. Applied.

Detailed experimental method is as follows. First, human-derived fibroblasts were dispensed into a 24 well plate for cell culture, and cultured in a medium containing 10% FBS (fetal bovine serum) for 24 hours at 37 ° C. and 5% CO _2. Wash twice with phosphate buffer saline (PBS). After treating the herbal extract obtained in Example 1-24 to 0.001% in a medium containing fibroblasts and 1% FBS cultured according to this method, an amino acid (2,3- ³ H proline) labeled with radioisotopes ) 2 μCi and 30 μg / ml of ascorbic acid were added, followed by incubation for 24 hours at 37 ° C. and 5% CO ₂ . After the completion of the culture, the cells and the culture medium were divided into 1/2, and the collagenase was treated in only one fraction, and the protein was precipitated with trichloroacetic acid (TCA) to measure the amount of collagen biosynthesized from the difference in radioactivity of the two fractions. . Collagen biosynthesis promoting effect of each sample is shown in Table 2 as a collagen increase rate to 100% of the control group without the extract. In addition, the experiment was performed in the same manner using 80% methanol extract dried centella asiatica extract and vitamin A retinol (retinol) instead of the extract of the example and the results are shown in Table 2 as a collagen increase rate.

sample Amount of biosynthesized collagen (cpm) ^* Collagen Growth Rate (%) Control 1465 ± 228 100 Example 1 3861 ± 187 225 Example 2 2873 ± 236 173 Example 3 2493 ± 170 153 Example 4 2911 ± 168 175 Example 5 2607 ± 132 159 Example 6 2854 ± 211 172 Example 7 3310 ± 221 196 Example 8 2626 ± 193 160 Example 9 2398 ± 172 148 Example 10 3348 ± 232 198 Example 11 2664 ± 192 162 Example 12 2569 ± 255 157 Example 13 2341 ± 159 145 Example 14 2759 ± 168 167 Example 15 2094 ± 178 132 Example 16 3044 ± 182 180 Example 17 3066 ± 194 180 Example 18 3348 ± 168 196 Example 19 3671 ± 245 215 Example 20 2474 ± 257 152 Example 21 2702 ± 243 164 Example 22 2968 ± 198 178 Example 23 2759 ± 177 167 Example 24 3196 ± 196 190 Centella asiatica extract 1992 ± 168 124 Vitamin A (Retinol) 2837 ± 228 172

* Repeat count: 3

As can be seen from the results in Table 2, each of the samples obtained in Examples 1-24 showed a very strong collagen biosynthesis promoting effect despite the crude extract. In addition, the known effect of promoting collagen biosynthesis of fibroblastsCentella asiaticaof It was confirmed that there is an excellent effect when compared to the extract dried, and compared to the retinol, vitamin A showed a similar or superior effect even in the state of the extract was found to be excellent collagen biosynthesis.

Experimental Example 2 Collagen Biosynthesis Effect at Cell Level According to Concentration

For the fibroblasts derived from humans cultured in the same manner as in Experimental Example 1, the collagen biosynthesis promoting effect according to the concentrations of the extracts of Examples 1, 9 and 17 was investigated.

sample Application concentration (%) Amount of biosynthesized collagen (cpm) ^* Collagen Growth Rate (%) Control - 1465 ± 228 100 Example 1 0.00001 3342 ± 219 179 0.001 4438 ± 259 225 0.1 5129 ± 275 254 Example 9 0.00001 2484 ± 247 143 0.001 3416 ± 191 182 0.1 4200 ± 188 215 Example 17 0.00001 2746 ± 205 154 0.001 3580 ± 211 189 0.1 3652 ± 253 192 Centella asiatica extract 0.00001 1664 ± 416 108 0.001 1992 ± 168 124 0.1 2434 ± 153 140 Vitamin A (Retinol) 0.00001 2189 ± 147 138 0.001 2837 ± 228 172 0.1 929 ± 101 72

* Repeat number: 3

As shown in the results of Table 3, Sinyi extract, white paper extract and clove extract have very strong collagen biosynthesis on cultured human fibroblasts, even at low concentrations compared to the known Centella asiatica extract. It can be seen that the effect is very high. In addition, collagen biosynthesis showed similar or superior efficacy even when compared to known vitamin A extract. In particular, vitamin A showed toxicity to cultured cells at a concentration of 0.1%, resulting in low collagen biosynthesis by killing cells, but the extract of the present invention did not exhibit this phenomenon.

Preparation Example 1-5 and Comparative Example 1

Sinyi extract, white paper extract and clove extract and each component shown in Table 4 was mixed in the ratio shown in Table 4 to prepare an external skin ointment.

1, 2, 3: extracts of Examples 5, 13 and 21, respectively

4: amount to be 100% by weight in total

Preparation Example 6-10 and Comparative Example 2

Sinyi extract, white paper extract and clove extract and each component shown in Table 5 was mixed in the ratio shown in Table 5 to prepare a cream in cosmetics.

1, 2, 3: extracts of Examples 5, 13 and 21, respectively

4: amount to be 100% by weight in total

Preparation Example 11-15 and Comparative Example 3

Essence extract, white paper extract and clove extract and each component shown in Table 6 was mixed in the ratio shown in Table 6 to prepare an essence in the cosmetic.

1, 2, 3: extracts of Examples 5, 13 and 21, respectively

4: amount to be 100% by weight in total

Preparation Example 16-20 and Comparative Example 4

Sinyi extract, white paper extract and clove extract and each component shown in Table 7 was mixed in the ratio shown in Table 7 to prepare a flexible cosmetics in cosmetics.

1, 2, 3: extracts of Examples 5, 13 and 21, respectively

4: amount to be 100% by weight in total

Preparation Example 21-25 and Comparative Example 5

Xinyi extract, white paper extract and clove extract and each component shown in Table 8 was mixed in the ratio shown in Table 8 to prepare a nutrient lotion in cosmetics.

1, 2, 3: extracts of Examples 5, 13 and 21, respectively

4: amount to be 100% by weight in total

Experimental Example 3 Wound Healing Effect in Rats

Using the samples prepared in Preparation Examples 1-25 and Comparative Examples 1-5, the wound healing effect on rats was tested in the following manner.

Five-week-old male rats were tested for wound healing effects using a tensile strength method that reflects the quality and amount of wound regeneration in the incision window. The dorsal hair of the rat was removed and then cut with a scalpel and the incision site was closed. Samples prepared in Preparation Examples 1-25 and Comparative Examples 1-5 were administered to the incision site for 0.5 days / cm 2 for 5 days five times daily. Six days later, the rats were used for slaughter, the skin of the wound was extracted, three skin specimens of 1 cm width orthogonal to the incision were made for each subject, and the tearing tension was measured using a rheometer. Using the measured tensile strength (g / cm) as an index of the strength of the regenerated collagen fibers, the results are shown in Table 9-13, and the rate of increase in the tensile strength is shown in Comparative Example 1-5 without adding the sample. The tensile strength at the time of application was expressed as relative strength.

Wound healing effect on rats of samples prepared in Preparation Example 1-5 and Comparative Example 1 Sample ^* Tensile Strength (g / cm) % Increase Comparative Example 1 550 100 Preparation Example 1 709 129 Preparation Example 2 698 127 Preparation Example 3 682 124 Preparation Example 4 720 131 Preparation Example 5 710 129

* Repeat number: 3

Wound healing effect on rats of samples prepared in Preparation Example 6-10 and Comparative Example 2 Sample ^* Tensile Strength (g / cm) % Increase Comparative Example 2 563 100 Preparation Example 6 622 112 Preparation Example 7 647 117 Preparation Example 8 662 120 Preparation Example 9 672 122 Preparation Example 10 657 119

* Repeat number: 3

Wound healing effect on rats of samples prepared in Preparation Examples 11-15 and Comparative Example 3 Sample ^* Tensile Strength (g / cm) % Increase Comparative Example 3 530 100 Preparation Example 11 663 121 Preparation Example 12 638 117 Preparation Example 13 688 125 Preparation Example 14 707 128 Preparation Example 15 599 113

* Repeat number: 3

Wound healing effect on rats of samples prepared in Preparation Examples 16-20 and Comparative Example 4 Sample ^* Tensile Strength (g / cm) % Increase Comparative Example 4 545 100 Preparation Example 16 647 118 Preparation Example 17 625 114 Preparation Example 18 619 113 Preparation Example 19 642 117 Preparation Example 20 596 109

* Repeat number: 3

Wound healing effect on rats of samples prepared in Preparation Examples 21-25 and Comparative Example 5 Sample ^* Tensile Strength (g / cm) % Increase Comparative Example 5 563 100 Preparation Example 21 617 111 Preparation Example 22 598 107 Preparation Example 23 586 105 Preparation Example 24 629 113 Preparation Example 25 595 107

* Repeat number: 3

From the results of Table 9-13, the ointment and cosmetics containing Sinyi extract, Baidu extract and / or clove extract prepared according to Preparation Examples 1-25 according to the present invention were excellent in wound healing effect of rats. In particular, the comparative experiments of Preparation Example 1-25 and Comparative Example 1-5 to which a large amount of god extract, white paper extract and cloves extract were applied showed more than 120% of wound healing promoting effect. It also did not show any side effects in the skin.

As a result of the experiment, it was found that the extract of Sinyi, white paper and clove extract of the present invention has excellent wound healing effect as a safe collagen biosynthesis promoter.

The skin composition containing the extract of Sinyi, white paper and clove extract of the present invention exhibits a powerful collagen synthesis promoting effect upon fibroblast treatment, has excellent wound healing effect, and does not cause any side effects, and is excellent in safety.

Claims (3)

Wrinkle healing skin composition comprising one or more extracts selected from the group consisting of shinyi extract, white paper extract and clove extract as a collagen synthesis promoter. The method of claim 1, The extract is wound, white paper and cloves, respectively, water, a lower alcohol having 1 to 4 carbon atoms, ethyl acetate, acetone and a nucleic acid is extracted using at least one mixed solvent selected from the group consisting of nucleic acids. The method of claim 1, The amount of the extract is wound healing skin composition of 0.0001-10.0% by weight of the total composition.