KR102193139B1 - A cosmetic composition comprising belutin - Google Patents

Abstract

The present invention relates to a cosmetic composition and the like containing beloutin, and by the present invention, it has been found that beloutin has an excellent whitening effect. Therefore, the beloutine of the present invention can be usefully used as a cosmetic composition for whitening in the field of cosmetic manufacturing.

Description

Cosmetic composition comprising belutin {A cosmetic composition comprising belutin}

The present invention relates to a cosmetic composition and the like containing veloutine.

Mistletoe (Mistletoe, Viscum album ) is a semi-parasitic plant that grows mainly by broadleaf trees such as oak and chestnut as a host, and has been used as a medicinal plant in East Asia and Europe, including Korea. In particular, mistletoe living in Europe is known to have excellent anticancer activity and is commercially marketed as an anticancer treatment/adjuvant, and in Korea, it is used as a medicine for toothache, hypertension, miscarriage, and low back pain in oriental medicine.

Regarding the whitening effect of mistletoe, Korean Patent Application Publication No. 2002-0080658 has reported the whitening effect of mistletoe extract, and Korean Patent Application Publication No. 10-2018-001232 has antioxidant and whitening activity of fractions fractionated from mistletoe extract. Has been disclosed.

However, up to now, a simple method for improving the whitening effect using the mistletoe extract has not been developed, and a method for converting the mistletoe extract into a non-glycoside for glycosides has not been developed. Therefore, there is a need to develop a simple and effective method for converting glycosides of mistletoe extract into non-glycosides.

Korean Patent Application Publication No. 10-2012-0085495 (2012.08.01.) Korean Patent Application Publication No. 10-2014-0147595 (2014.12.30.) Korean Patent Application Publication No. 10-2014-0087051 (2014.07.08)

The present invention provides a cosmetic composition comprising veloutine.

In order to achieve the object of the present invention, the present invention provides a cosmetic composition containing veloutine.

In one embodiment of the present invention, the cosmetic may be for whitening.

In one embodiment of the present invention, the composition may have an ability to inhibit tyrosinase activity.

In one embodiment of the present invention, the beloutine may be derived from mistletoe extract.

By the present invention, it was found that beloutine has an excellent whitening effect. Therefore, the beloutine of the present invention can be usefully used as a cosmetic composition for whitening in the field of cosmetic manufacturing.

1 is a result of analysis of bisquineoside III, which is a glycoside of raw mistletoe, by HPLC.
2 is a result of analyzing homoflavoyadolinin B, which is a glycoside of raw mistletoe, by HPLC.
3 is a result of analyzing homoeriodictiol, a non-glycoside of raw mistletoe, by HPLC.
4 is a result of analysis by HPLC analyzing beloutine (Comparative Example 6), which is a non-glycoside of raw mistletoe.
5 is a result of HPLC analysis of a lyophilisate (Comparative Example 1) in which the mistletoe extract was not heat-treated in 50% MeOH.
6 is a result of analyzing glycosides and non-glycosides components in a microwave irradiated product (Example 1) subjected to microwave processing of a mistletoe extract in 1% AcOH at 100° C. and 100 W for 1 hour by HPLC.
7 is a result of analyzing glycosides and non-glycosides components in a microwave irradiated product (Example 2) in which mistletoe extract was microwaved at 120° C. and 100 W for 1 hour in 1% AcOH by HPLC.
8 is a result of analyzing glycosides and non-glycosides components in a microwave irradiated product (Example 3) subjected to microwave processing of mistletoe extract in 1% AcOH at 120° C. and 100 W for 3 hours by HPLC.
9 is a result of analyzing glycosides and non-glycoside components in a microwave irradiated product (Example 4) subjected to microwave processing at 120° C. and 100 W in 10% AcOH of mistletoe extract for 1 hour by HPLC.
10 is a result of analyzing glycosides and non-glycoside components in a microwave irradiated product (Example 5) subjected to microwave processing at 120° C. and 200 W in 10% AcOH of mistletoe extract for 1 hour by HPLC.
11 is a result of analyzing glycosides and non-glycosides components in a microwave irradiated product (Example 6) subjected to microwave processing at 120° C. and 100 W in 50% AcOH of mistletoe extract for 1 hour by HPLC.
12 is a result of analysis of glycosides and non-glycoside components in a microwave irradiated product (Example 7) subjected to microwave processing at 120° C. and 100 W in 100% AcOH of mistletoe extract for 1 hour by HPLC.
13 is a result of analyzing glycosides and non-glycoside components in a microwave irradiated product (Example 8) subjected to microwave processing at 120° C. and 100 W in 100% AcOH of mistletoe extract for 30 minutes by HPLC.
14 is a result of analyzing a processed product (Comparative Example 2) obtained by heat treatment of a mistletoe extract in 100% AcOH at 120° C. for 12 hours by HPLC.
FIG. 15 is a result of analyzing a microwave irradiation product (Comparative Example 3) obtained by microwave processing at 150° C. and 100 W for 30 minutes in 1% AcOH of mistletoe extract by HPLC.
16 is a result of measuring the ability to inhibit tyrosinase activity, which is an evaluation of whitening activity, with respect to the resultant of the mistletoe extract according to the microwave irradiation conditions.
17 is a result of measuring the ability to inhibit tyrosinase activity, which is an evaluation of whitening activity, for each single component of a mistletoe extract.

The present invention provides a method for converting glycosides into non-glycosides comprising the step of irradiating microwaves in the presence of acetic acid in mistletoe extract. The mistletoe ( Viscum album L. var. coloratum) is naturally dried, and then 70% (v/v) ethanol is added to the mistletoe outpost, and extracted three times for 1 to 5 hours at room temperature. The obtained 70% ethanol extract is collected, filtered, and concentrated under reduced pressure to obtain a dried mistletoe extract.

In one embodiment, the acetic acid is 1 to 100% (v/v), preferably 10 to 100% (v/v), most preferably 50 to 100% (v/v). I can.

In one embodiment, it may include that the microwave irradiation is carried out under a temperature of 80 to 140 °C, preferably 95 to 125 °C.

In one embodiment, it may include that the irradiation of the microwave is performed under a power amount of 100 to 200 W (watt).

In one embodiment, it may include that the microwave irradiation is performed for 30 minutes to 3 hours, preferably 45 minutes to 2 hours.

In one embodiment, it may include that the microwave irradiation is performed under a pressure of 5 to 50, preferably 10 to 30 atmospheres.

In one embodiment, the mistletoe extract is irradiated with microwaves for 30 minutes to 3 hours under 1 to 100% (v/v) acetic acid, irradiation temperature 80 to 140 °C, 100 to 200 W (watt) power and 5 to 50 atmospheres It may include a method of converting a glycoside to a non-glycoside comprising the step of.

In one embodiment, in the mistletoe extract 100% (v / v) acetic acid, irradiation temperature 120 ℃, 100 W (watts) comprising the step of irradiating microwaves for 1 hour under 20 atmospheres, as a non-glycoside of glycoside It may include a conversion method of.

In addition, the present invention provides a mistletoe extract containing a non-glycoside obtained by the above conversion method.

In one embodiment, the non-glycoside may be velutin or homoeriodictyol. The non-glycosides, beloutin and homoeriodicthiol, are non-glycosides converted from Homoflavoyadorinin B and Viscumneoside III through deglycosylation.

In addition, the present invention provides a cosmetic composition for whitening comprising a mistletoe extract containing a non-glycoside obtained by the conversion method. As a result of measuring tyrosinase inhibitory activity, which is a whitening efficacy evaluation index, using the mistletoe extract, the inhibitory effect was best when 100% AcOH, which was all converted to non-glycoside, was used as a solvent (see FIG. 16). In addition, as for the tyrosinase inhibitory effect of each single component, mistletoe non-glycosides showed significantly increased efficacy than glycosides, and showed an inhibitory effect similar to that of ascorbic acid, a positive control (see FIG. 17).

Hereinafter, the present invention will be described in more detail through Preparation Examples, Examples and Test Examples. However, the following Preparation Examples, Examples, and Test Examples are for illustrating the present invention, and the scope of the present invention is not limited thereto.

Preparation Example 1. Preparation of mistletoe extract

To the outpost of naturally dried mistletoe ( Viscum album L. var. coloratum ) (30 g, purchased from Sunil Mulsan), 300 mL of 70% (v/v) ethanol was added, and extracted three times for 3 hours at room temperature. The obtained 70% ethanol extract was collected, filtered, and concentrated under reduced pressure to obtain 2.6 g of a dried mistletoe extract.

Examples 1 to 8. Preparation of microwave-irradiated processed mistletoe extract

The 70% ethanol extract obtained in Example 1 was subjected to microwave irradiation under the conditions of Table 1. Specifically, 1 mL of water to add 200 mg of dried mistletoe extract to a 10 mL container of a microwave irradiator (CEM, model number 908005) was adjusted according to the AcOH content (v/v) in Table 1 below, and the container was sealed. In one state, microwaves were irradiated according to the conditions of temperature, power amount, and irradiation time in Table 1 below, and then freeze-dried to obtain a microwave-irradiated workpiece. The pressure during microwave irradiation was 20 atmospheres.

Solvent (1 mL) Temperature (℃) Wattage (W) Irradiation time (h) Example 1 1% AcOH 100 100 One Example 2 1% AcOH 120 100 One Example 3 1% AcOH 120 100 3 Example 4 10% AcOH 120 100 One Example 5 10% AcOH 120 200 One Example 6 50% AcOH 120 100 One Example 7 100% AcOH 120 100 One Example 8 100% AcOH 120 100 0.5

Comparative Examples 1 and 2. Preparation of processed mistletoe extract without microwave irradiation

In order to compare the component differences between the microwave irradiated products prepared according to Examples 1 to 8 and the mistletoe extract heat-treated according to the conventional method, the mistletoe extract without heating, the simple heat-treated extract and the processed extract subjected to microwave irradiation at 150°C Was prepared. 200 mg of the 70% ethanol extract obtained in Preparation Example 1 was added to 1 mL of 50% methanol and not heat treated (Comparative Example 1), and 200 mg of the 70% ethanol extract was added to 1 mL of 100% AcOH, and then simply about 120 °C A simple heat treatment extract (Comparative Example 2) obtained by lyophilizing the processed mistletoe extract heated through reflux extraction for 12 hours. In addition, 200 mg of 70% ethanol extract was added to 1 mL of 1% AcOH to obtain a processed product (Comparative Example 3) irradiated with microwaves at 150° C. for 30 minutes.

<Test Example>

1. Preparation of Comparative Substance-Synthesis of single component contained in mistletoe extract

Bisquineoside III and Homoflavoyadorin B, known as active ingredients of mistletoe, used a compound isolated through Korean Patent Laid-Open Patent No. 10-2018-0001232, and homoeriodicthiol, a non-glycoside of each substance, and Belloutin was synthesized using a combination of previously known synthetic methods.

2. Analysis of non-glycosides in processed mistletoe extract

Using the comparative material synthesized above as a control, the method of the literature (Lee et al., Korean J. Plant Res. 2016, 29, 690) by HPLC, the glycosides and non-glycoside components of each of the processed mistletoe extracts obtained in Examples 1 to 8 and Comparative Examples 1 to 3 were analyzed. The analysis was repeated three times to confirm reproducibility.

device YL-9100 (Younglin instrument, Korea) column TC-C18 column (250 mm × 4.6 mm, particle size 5 μm, Agilent, USA) menstruum 0.1% formic acid (A), MeOH (B) Mobile phase concentration gradient 0-20 min, 40-52% B; 20 to 40 min, 52 to 80% B; 5 min, 40% B Mobile phase speed 0.8 mL/min Column temperature 30 ℃ Detector UV 270 nm

As a result of the experiment, as a result of analyzing each component separated from the column by HPLC analysis for the processed mistletoe extract, the chromatograms of FIGS. 1 to 14 were obtained.

In the results of FIGS. 6 to 8, bisquineoside III and homoflavoyadorin B, which are glycosides contained in mistletoe, are non-glycosides as the temperature increases during microwave treatment at 100°C or 120°C in 1% AcOH solvent. Each was converted to, but mainly obtained peaks, which were not completely non-glycosylated, but were expected to be partially departed from sugar. Although the microwave irradiation time was increased to 3 hours, complete conversion to non-glycosides did not occur.

In the results of FIGS. 9 to 13, it was confirmed that as the content of AcOH increased, the peak expected to be partially desorbed from the sugar decreased, and the non-glycoside peak increased. In particular, in the case of 100% AcOH, this tendency was best shown (see FIGS. 12 and 13). However, even if the amount of power was increased, the conversion of non-glycosides was not significantly affected. When the reaction time was reduced to 30 minutes, the conversion to non-glycoside was not completely achieved.

In the results of FIG. 14, when the mistletoe was simply heated, all of the glycoside peaks disappeared, but many other peaks were generated in addition to the non-glycoside peak, and it seemed that the non-glycoside was not produced as the main product.

In the results of FIG. 15, when mistletoe was added to 1 mL of 1% AcOH and irradiated with microwave for 30 minutes at 150° C., not only the glycoside peak but also the peak of the extract were hardly seen. It has been shown that most of the compounds are decomposed when done.

Therefore, when the mistletoe extract is irradiated with microwave, it is possible to more efficiently convert the main glycosides of mistletoe, bisquineoside III and homoflavoyadorin B, into homoeriodicthiol and veloutine, which are highly effective non-glycosides. there was.

3. Evaluation of whitening activity of processed mistletoe, mistletoe glycosides and non-glycosides

For the solvent fractions of the obtained mistletoe extract, the ability to inhibit tyrosinase enzyme activity, which is a representative method of a whitening efficacy evaluation test, was measured as follows. To a 96-well plate, 110 μL of 0.1 M phosphate buffer (pH 6.8) and 30 μL of samples prepared at concentrations of 0.2 mg/mL and 0.5 mg/mL were added, and 20 μL of tyrosinase (250 U/mL) was added. It was added and reacted at 35°C for 5 minutes. Thereafter, 40 μL of 5 mM L-DOPA (L-3,4-dihydroxy-L-phenylalanine) solution was added and reacted at 35° C. for 5 to 10 minutes, and then at 475 nm, an ELISA reader (enzymelinked immunosorbent assay, ELISA reader) Absorbance was measured by furnace. The ability to inhibit tyrosinase was calculated by Equation 1 below, and the results are shown in FIGS. 16 and 17. In this test, the mean ± standard deviation value was calculated using SPSS (v.24 for Windows) for the results obtained by measuring in three repetitions, and the significance was tested through the Bonferroni test. ^* p<0.05 was considered significant.

[Equation 1]

Tyrosinase inhibitory ability (%) = {1-(Sample absorbance/Control material absorbance)} × 100

As shown in Figure 16, the inhibitory effect of tyrosinase on the mistletoe and processed mistletoe extract was the best when 100% AcOH converted to non-glycoside was used as a solvent, and 1% AcOH was used, When only part of the sugar was released, the inhibitory effect was not better than when it was not.

FIG. 17 shows the tyrosinase inhibitory effect of each single component, and the tyrosinase inhibitory effect was increased by 10% or more in the case of non-glycosides than in the case of glycosides. In addition, in the case of the non-glycoside, it showed an inhibitory effect at a level similar to that of ascorbic acid as a control.

Claims (4)

Whitening cosmetic composition containing beloutine.
delete The cosmetic composition of claim 1, wherein the composition has an ability to inhibit tyrosinase activity.
The cosmetic composition according to claim 1, wherein the beloutine is derived from mistletoe extract.

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