TWI826254B - Whitening composition and use thereof - Google Patents

Abstract

A whitening composition is used to inhibit the production of melanin. The whitening composition includes an extract of Sarcandra glabra, an extract of Rosmarinus officinalisand an extract of Andrographis paniculata. All of the extract of Sarcandra glabra, the extract of Rosmarinus officinalisand the extract of Andrographis paniculateare manufactured by extracting a sample of Sarcandra glabra, a sample of Rosmarinus officinalisand a sample of Andrographis paniculate, respectively, at a pressure ranging from 1 Kgf/cm ²to 1.2 Kgf/cm ²and a temperature ranging from 121°C to 135°C using an aqueous phosphate buffer saline (PBS) solution as an extractant. The invention also relates to a use of the whitening composition.

Description

Whitening compositions and uses thereof

The present invention relates to a composition, especially a whitening composition. The present invention also relates to the use of the whitening composition.

It is human nature to love beauty, and this is true at all times and in all countries. However, for Orientals, the concept of whitening to cover up three ugliness has far-reaching influence, which leads Orientals to love whitening more than others. Therefore, the output value of whitening and skin care products remains high in the market, which has also led to the development of many new raw materials.

The human body turns black when exposed to sunlight, which is the body's defense mechanism. After a series of biochemical reactions, melanin is produced as a metabolic product, which is also the main substance that causes skin darkening. In short, melanocytes contain tyrosin. When tyrosinase in melanocytes is activated, it will promote the conversion of tyrosine into dopa (3,4-dihydroxyphenylalanine, referred to as DOPA). ), and dopa will be further converted into dopaquinone, eventually forming melanin such as eumelanin and pheomelanin. Therefore, this series of biochemical reactions can be attributed to the reaction of tyrosinase in melanocytes.

In other words, if we can find a way to inhibit the activity of tyrosinase in melanocytes, we can effectively reduce the production of melanin. Therefore, many researchers and developers focus on finding natural ingredients from land plants or marine organisms that can whiten, lighten skin tone, and inhibit melanin production, hoping to develop effective ingredients with better efficacy.

In order to solve the above problems, the main object of the present invention is a whitening composition that can effectively inhibit the activity of tyrosinase in melanocytes.

A secondary object of the present invention is to provide the use of a whitening composition, which is to apply the whitening composition of the present invention to prepare whitening preparations.

The use of the quantifier "a" or "an" in the elements and components described throughout the present invention is only for convenience of use and to provide a common sense of the scope of the present invention; in the present invention, it should be interpreted as including one or at least one, and single The concept of also includes the plural unless it is obvious that something else is meant.

The whitening composition of the present invention may include: a herbaceous coral extract; a rosemary extract; and an Andrographis paniculata extract, and the herbaceous coral extract, the rosemary extract and the Andrographis paniculata extract can be combined with one Phosphate buffered saline solution was used as ^the extraction agent, and a sample of Coral Coral, a sample of Rosemary and A sample of Andrographis paniculata was obtained.

Accordingly, the whitening composition of the present invention, composed of the grass coral extract, the rosemary extract and the Andrographis paniculata extract, can not only effectively inhibit the activity of tyrosinase, but also block the production of melanin. Relevant message transmission pathways can further reduce the production of melanin. Therefore, the whitening composition can be used to prepare whitening preparations for administration to a desired individual to inhibit the production of melanin in the melanocytes of the desired individual. Efficacy of the present invention.

The whitening composition of the present invention, wherein the phosphate buffered physiological saline solution can include sodium chloride with a concentration between 0.12 and 0.14 M and sodium dihydrogen phosphate with a concentration between 0.008 and 0.012 M, and the phosphoric acid The pH of the salt-buffered saline solution may be neutral. In this way, the grass coral extract, rosemary extract and Andrographis paniculata extract obtained by using phosphate buffered saline as the extraction agent have better inhibitory activity against tyrosinase.

In the whitening composition of the present invention, after obtaining the Herba Coral extract, the Rosemary extract and the Andrographis paniculata extract respectively, the Herba Coral extract, the Rosemary extract and the Andrographis paniculata extract can be mixed. to form the whitening composition. In this way, by separately extracting the grass coral sample, the rosemary sample and the andrographis paniculata sample, the extraction conditions can be adjusted respectively according to the status of the grass coral sample, the rosemary sample and the andrographis paniculata sample, thereby improving the obtained Quality of Grass Coral Extract, Rosemary Extract and Andrographis Paniculata Extract.

The whitening composition of the present invention, wherein the whitening composition may comprise 15 to 55% of Coral grass extract, 30 to 55% of Rosemary extract and 15 to 55% of Andrographis paniculata extract by weight percentage; Preferably, it may include 33.3% of Coral grass extract, 50% of Rosemary extract and 16.7% of Andrographis paniculata extract by weight. In this way, the whitening composition has better tyrosinase inhibitory activity through the specific ratio of the grass coral extract, the rosemary extract and the Andrographis paniculata extract.

The use of the whitening composition of the present invention is applied to the preparation of whitening preparations, wherein the whitening composition is the aforementioned whitening composition, and is administered to a desired individual to inhibit the production of melanocytes in the desired individual. The production of melanin.

Accordingly, the use of the whitening composition of the present invention can further achieve the effect of effectively inhibiting the production of melanin by using the aforementioned whitening composition.

The use of the whitening composition of the present invention, wherein the whitening composition can be administered to the desired individual by smearing or spraying. In this way, the whitening composition can be conveniently administered to the desired individual through the selected administration method.

The use of the whitening composition of the present invention, wherein the whitening composition can be administered to the desired individual at a dose of 200 to 500 μg/cm ² . In this way, by selecting the dosage, the whitening composition can exert a better whitening effect.

In order to make the above and other objects, features and advantages of the present invention more obvious and understandable, preferred embodiments of the present invention are illustrated in detail below and described in detail with reference to the accompanying drawings.

The whitening composition according to an embodiment of the present invention may include: a Coral Coral extract, a rosemary extract and an Andrographis paniculata extract, so that through the Coral Coral extract, the Rosemary extract and the The combined effect of Andrographis paniculata extract can effectively inhibit the activity of tyrosinase and block the signal transduction pathway related to melanin synthesis, thereby reducing the production of melanin and reducing the melanin content in melanocytes.

In detail, the method for manufacturing the whitening composition can be as shown in Figure 1, including: a step S1 of preparing a herbaceous extract, a step S2 of preparing a rosemary extract, and a step S2 of preparing an Andrographis paniculata extract. Step S3, to prepare the Coral grass extract, the rosemary extract and the Andrographis paniculata extract respectively. And after obtaining the Herba Coral extract, the Rosemary extract and the Andrographis paniculata extract respectively, the worker can then mix the Herba Coral extract, the rosemary extract and the Andrographis paniculata through a preparation step S4. extract to obtain the whitening composition.

In the preparation step S1 of the grass coral extract, the worker extracts a grass coral sample with an extraction agent. The grass coral sample can be a plant of the genus Sarcandra in the Chloranthaceae family (ie, Sarcandra glabra ), and preferably can be the whole plant of the aforementioned plant. For example, the grass coral extract can be prepared by a method including the following steps: providing a sample as a step S11, an extraction as a step S12, and a concentration as a step S13.

In the sample providing sub-step S11, the worker can directly perform the extraction sub-step S12 after obtaining the grass coral sample, or the worker can pre-dry the grass coral sample before performing the extraction sub-step S12, for example Drying reduces the water content of the grass coral sample to 3-10%; in addition, the grass coral sample can also be crushed in advance, for example, the particle size of the grass coral sample can be between 5-20 nm to increase the The contact surface area of the extraction agent used in the extraction sub-step S12 thereby improves the subsequent extraction efficiency.

In the extraction sub-step S12, the worker can use a phosphate buffer saline (PBS) solution as an extraction agent to extract the grass coral sample. For example, the phosphate buffered saline solution may include an aqueous solution of sodium chloride (NaCl, sodium chloride) and sodium dihydrogen phosphate (NaH ₂ PO ₄ , sodium dihydrogen phosphate), and the pH value of the aqueous solution is approximately For neutral. For example, the phosphate buffered saline solution may contain sodium chloride at a concentration of 0.12-0.14 M and sodium dihydrogen phosphate at a concentration of 0.008-0.012 M. Preferably, it may contain chloride at a concentration of 0.13 M. Sodium and sodium dihydrogen phosphate with a concentration of 0.01 M, and the pH value of the phosphate buffered saline solution is 7.0.

In this embodiment, 200 ml of the extraction agent can be mixed with every 200 grams of grass coral sample to obtain a mixture, and then the mixture is mixed with a mixture of 1 to 1.2 Kgf/cm ² (for example, 1.033 Kgf/cm ² ). Extraction is performed under pressure and at a temperature between 121 and 135°C (for example, 127°C), and the extraction time can be between 20 and 30 minutes (for example, 30 minutes), so that the grass coral sample is rich in active ingredients. It can be completely dissolved in the extractant, so that a crude coral extract can be obtained.

In the concentration sub-step S13, the worker can freeze-dry the crude coral extract after obtaining the crude coral extract to obtain the coral grass extract. Through this procedure, the system can The active ingredients of the grass coral extract are more concentrated, so only a small amount of the grass coral extract is needed to achieve the best effect. In this example, a high-speed centrifuge was used to centrifuge at 12,000 rpm for 20 minutes to precipitate the residues in the rough coral extract, and then the residues were removed, and then the mixture was centrifuged at a temperature between -50 and -80°C. The obtained supernatant liquid is freeze-dried at a temperature between 100% and 200% to obtain dry powder, and the grass coral extract can be obtained.

Furthermore, in the rosemary extract preparation step S2 and the Andrographis paniculata extract preparation step S3, the workers extracted a rosemary sample and an Andrographis paniculata sample with the extractant respectively. The rosemary sample can be a plant of the genus Rosmarinus (Lamiaceae) [i.e., rosemary ( Rosmarinus officinalis )], preferably the whole plant of the aforementioned plant, and the Andrographis paniculata sample can be For a plant of the genus Andrographis in the family Acanthaceae (ie, Andrographis paniculata ), it is preferably the whole plant of the aforementioned plant. For example, the rosemary extract can be prepared by a method including the following steps: providing a sample in step S21, an extraction in step S22, and a concentration in step S23. The Andrographis paniculata extract can be prepared by including It is prepared by the following steps: providing a sample in step S31, extracting in step S32, and concentrating in step S33.

In the sample providing sub-step S21 (or the sample providing sub-step S31), the worker may directly perform the extraction sub-step S22 after obtaining the rosemary sample (or the Andrographis paniculata sample), or may also perform the extraction sub-step S22 as described above. As described above, before performing the extraction sub-step S22, dry or pulverize in advance to reduce the water content of the rosemary sample (or the Andrographis paniculata sample) to 3-10%, or make the rosemary sample (or The particle size of the Andrographis paniculata sample) can be between 5 and 20 nm.

In the extraction sub-step S22 (or the extraction sub-step S32), the worker can use the phosphate buffered saline solution as an extraction agent to extract the rosemary sample (or the Andrographis paniculata sample). The phosphate buffered physiological saline solution can be the same as the formula in the extraction sub-step S12 of the grass coral extract preparation step S1. The craftsman can also use the rosemary sample (or the Andrographis paniculata sample) and the grass coral. Self-adjustment due to differences in the state of the sample is understandable to those with ordinary knowledge in the technical field to which the present invention belongs, and will not be described in detail here.

In this embodiment, every 100 grams of rosemary sample (or Andrographis paniculata sample) can be mixed with 200 ml of the extraction agent to obtain a mixture, and then in the extraction sub-step S12 of the grass coral extract preparation step S1 Extraction is carried out under the pressure and temperature range described in, and the extraction time can also be roughly the same as the extraction sub-step S12 of the grass coral extract preparation step S1, to obtain a rosemary crude extract (or an Andrographis paniculata crude extract). ).

In the concentration sub-step S23 (or the concentration sub-step S33), the worker can, after obtaining the rosemary crude extract (or the Andrographis paniculata crude extract), add the rosemary crude extract (or the Andrographis paniculata crude extract). Andrographis paniculata crude extract) is freeze-dried to obtain the rosemary extract (or the Andrographis paniculata extract). For example, a high-speed centrifuge can be used to centrifuge at 12,000 rpm for 20 minutes to obtain the rosemary crude extract. After the residues in (or the Andrographis paniculata crude extract) are precipitated, the residues are removed, and then the resulting supernatant is freeze-dried at a temperature between -50 and -80°C to obtain a dry powder, i.e. This rosemary extract (or this andrographis paniculata extract) is available.

Finally, in the preparation step S4, the worker can mix the aforementioned grass coral extract, rosemary extract and andrographis paniculata extract according to the required ratio to obtain the whitening composition. For example, the whitening composition may include 15% to 55% of Coral grass extract, 30% to 55% of rosemary extract, and 15% to 55% of Andrographis paniculata extract by weight. Preferably, it may include: The weight percentages are 33.3% Herba Coral extract, 50% Rosemary extract and 16.7% Andrographis paniculata extract.

It is worth noting that although workers can follow the method for manufacturing the whitening composition as shown in Figure 1, in the preparation step S1 of the grass coral extract and the preparation step S2 of the rosemary extract respectively And the Andrographis paniculata extract preparation step S3 is to respectively prepare the grass coral extract, the rosemary extract and the Andrographis paniculata extract, and then continue to obtain the whitening composition through the preparation step S4. However, when the various parameters used are roughly the same, workers can also first mix the grass coral sample, the rosemary sample and the andrographis paniculata sample, and then use the extractant to extract the grass coral sample, the rosemary sample sample and a mixed sample of the Andrographis paniculata sample (for example, when it is expected to obtain a whitening composition containing 33.3% Herba Coral extract, 50% Rosemary extract and 16.7% Andrographis paniculata extract by weight percentage, The mixed sample may include 33.3% of the grass coral sample, 50% of the rosemary sample and 16.7% of the Andrographis paniculata sample by weight), and the whitening composition can also be obtained by continuing to concentrate, which can improve the preparation of the whitening composition. The convenience of the product thereby reduces the preparation cost of the whitening composition.

The whitening composition of the present invention can inhibit the expression and activity of tyrosinase in melanocytes, thereby reducing the melanin content in melanocytes. Therefore, it can be used to prepare whitening preparations. The whitening composition can be combined with at least one medicine. A scientifically acceptable carrier is used to form the whitening preparation, or other active ingredients that reduce the melanin content in melanocytes are additionally added to the whitening preparation to be co-administered to a desired individual (for example, 200 to 500 μg can be used) /cm ² is applied to the skin of the desired individual), the carrier system is an excipient or an additive, preferably it can jointly form various types such as liquid, emulsion, cream or gel. The preparation is in a state that can be easily used by the desired individual.

In order to confirm that the whitening composition of the present invention can effectively inhibit the activity of tyrosinase, the following tests were conducted:

(A) Tyrosinase inhibition test (I)

This test is based on mixing the grass coral extract, the rosemary extract and the Andrographis paniculata extract according to the weight percentages shown in Table 1 to obtain whitening compositions of groups A6 to A8 (the grass coral extract , the total concentration of the rosemary extract and the Andrographis paniculata extract is 150 μg/mL); in addition, the grass coral extract, rosemary extract and Andrographis paniculata extract with a concentration of 150 μg/mL are respectively used as A3 ~Samples to be tested in group A5. In addition, Group A0 of this test is the blank group, that is, the group in which no extract and vitamin C are added (only tyrosinase and dopa-containing matrix solution are added), and Group A1 is the negative control group. , that is, the group with only dopa-containing matrix fluid. Group A2 is the positive control group. No extract is added, but vitamin C (40 μg/mL), tyrosinase and dopa-containing solution are added. matrix fluid.

Table 1 shows the samples to be tested in each group of this test. Group Weight percentage (%) Grass Coral Extract rosemary extract Andrographis paniculata extract Group A0 - - - Group A1 - - - Group A2 - - - Group A3 100% - - Group A4 - 100% - Group A5 - - 100% Group A6 50.0% 33.3% 16.7% Group A7 33.3% 50.0% 16.7% Group A8 16.7% 33.3% 50.0%

Mix the sample to be tested (60 μL) as described above with tyrosinase (3 μL). After reacting for 5 minutes, add matrix solution (DOPA 10 mM; 540 μL) and react at 37°C for 20 minutes. , allowing tyrosinase to convert dopa in the matrix solution into dopaquinone. Finally, measure the absorbance value of each group of reaction solutions at a wavelength of 450 nm, and calculate the relative absorbance value of each group of reaction solutions using the following formula 1, which can be converted to obtain the relative tyrosinase activity of each group of samples to be tested. (Formula 1)

Please refer to Table 1 and Figure 2. Adding alone the grass coral extract, the rosemary extract or the Andrographis paniculata extract can effectively inhibit the activity of tyrosinase (Groups A3 to A5; *: P < 0.05, compared with group A0), and adding the whitening composition containing the grass coral extract, the rosemary extract and the andrographis paniculata extract can also effectively inhibit the activity of tyrosinase (group A6~ Group A8; *: P <0.05, compared with group A0).

It is worth noting that, please refer to Table 1 and Figure 2 again, compared with groups A3 to A5 in which the grass coral extract, the rosemary extract or the Andrographis paniculata extract were added separately, the addition of The whitening composition of the grass coral extract, the rosemary extract and the Andrographis paniculata extract can more effectively inhibit the activity of tyrosinase (groups A6 to A8; @: P <0.05, compared with group A3 ; $: P < 0.05, compared with group A4; %: P < 0.05, compared with group A5), showing that the whitening composition uses the grass coral extract, the rosemary extract and the andrographis paniculata The combined effect of the extracts has better tyrosinase inhibitory activity.

In particular, please refer to Table 1 and Figure 2, which are groups A6 and A8 that also add the whitening composition containing the grass coral extract, the rosemary extract and the Andrographis paniculata extract. In comparison, Group A7, which added a whitening composition containing 33.3% of Coral grass extract, 50.0% of Rosemary extract and 16.7% of Andrographis paniculata extract by weight, was more effective in inhibiting tyrosinase. Activity (Groups A6, A8; &: P < 0.05, compared with Group A7), showing that the whitening composition containing the aforementioned specific ratio is composed of the grass coral extract, the rosemary extract and the Andrographis paniculata extract. The specific ratio has better tyrosinase inhibitory activity, so subsequent tests were conducted using the whitening composition in the specific ratio mentioned above.

(B) Tyrosinase inhibition test (II)

Please refer to Table 2. The whitening compositions of Groups B3 to B5 of this trial also include 33.3% by weight of grass coral extract, 50.0% of rosemary extract and 16.7% of Andrographis paniculata extract. , but it should be noted that the grass coral extract, rosemary extract and Andrographis paniculata extract in the whitening composition of Group B3 are all extracted using 95% ethanol aqueous solution as the extraction agent; the whitening composition of Group B4 The grass coral extract, rosemary extract and andrographis paniculata extract in the composition are all extracted using 99.9% methanol aqueous solution as the extraction agent; and the grass coral extract, rosemary extract in the whitening composition of Group B5 Both the aromatic extract and the Andrographis paniculata extract were extracted using the phosphate buffered saline solution as the extraction agent. In addition, Group B0 of this experiment is the blank group as mentioned above, Group B1 is the negative control group as mentioned above, and Group B2 is the positive control group as mentioned above, and each group is calculated according to the following formula 2 The relative absorbance value of the reaction solution can be converted into the relative tyrosinase activity of each group of samples to be tested. (Formula 2)

Table 2 shows the samples to be tested in each group of this test. Group Extractant Grass Coral Extract (33.3%) Rosemary extract (50.0%) Andrographis paniculata extract (16.7%) Group B0 - - - Group B1 - - - Group B2 - - - Group B3 ethanol aqueous solution ethanol aqueous solution ethanol aqueous solution Group B4 Methanol aqueous solution Methanol aqueous solution Methanol aqueous solution Group B5 Phosphate buffered saline solution Phosphate buffered saline solution Phosphate buffered saline solution

Please refer to Table 2 and Figure 3 for the whitening composition of Group B3 extracted with 95% ethanol aqueous solution as the extractant, or with the B4 whitening composition extracted with 99.9% methanol aqueous solution as the extractant. Compared with the whitening compositions of Group B5, the whitening compositions of Group B5 extracted using the phosphate buffered saline solution as the extraction agent can more effectively inhibit the activity of tyrosinase (Groups B3 and B4; $: P <0.05 , compared with Group B5), showing that the whitening composition can more easily extract the properties from the grass coral sample, the rosemary sample and the Andrographis paniculata sample by using the phosphate buffered saline solution as the extraction agent. Active ingredient with tyrosinase inhibitory activity.

(C)Melanin content measurement

Please refer to Table 3. The whitening composition of Group C3 of this trial contains 33.3% Coral grass extract, 50.0% Rosemary extract and 16.7% Andrographis paniculata extract by weight percentage, and the The grass coral extract, rosemary extract and Andrographis paniculata extract in the whitening composition are all extracted using the phosphate buffered physiological saline solution as the extraction agent. In addition, the C0 group of this experiment is the blank group as mentioned above, the C1 group is the negative control group as mentioned above, and the C2 group is the positive control group as mentioned above.

Table 3 shows the samples to be tested in each group of this test. Group Weight percentage (%) Grass Coral Extract rosemary extract Andrographis paniculata extract Group C0 - - - Group C1 - - - Group C2 - - - Group C3 33.3% 50.0% 16.7%

This test uses mouse melanoma B16-F10 cells as a model cell line. The test samples shown in Table 3 are added to the model cell line, and the model cell line is collected after 24 hours of reaction. Next, add sodium hydroxide (NaOH) aqueous solution (1 N, 0.2 mL) to the collected model cell line, and react at 80°C for 2 hours to dissolve the melanin in the model cell line. Centrifuge at 12,000 rpm for 30 minutes and collect the supernatant. Continue to measure the absorbance value of the supernatant in each group at a wavelength of 405 nm. Use the following formula 3 to calculate the relative absorbance value of the supernatant in each group, which can be converted. Know the relative melanin content of each group of samples to be tested. (Formula 3)

Please refer to Table 3 and Figure 4. The C3 whitening composition extracted using the phosphate buffered saline solution as the extraction agent can effectively reduce the melanin content in melanocytes, and the effect is the same as that of the positive control group. Vitamin C is similar.

(D) Effect on message transmission pathways related to melanin synthesis

Please refer to Table 4. The whitening composition of Group D1 of this trial contains 33.3% of Coral grass extract, 50.0% of Rosemary extract and 16.7% of Andrographis paniculata extract by weight, and the The grass coral extract, rosemary extract and Andrographis paniculata extract in the whitening composition are all extracted using the phosphate buffered physiological saline solution as the extraction agent. In addition, the D0 group in this test is the blank group as mentioned above.

Table 4 shows the samples to be tested in each group of this test. Group Weight percentage (%) Grass Coral Extract rosemary extract Andrographis paniculata extract Group D0 - - - Group D1 33.3% 50.0% 16.7%

This experiment also uses mouse melanoma B16-F10 cells as the model cell line. The test samples shown in Table 4 are added to the model cell line, and the model cell line is collected after 24 hours of reaction. Next, after lysing the model cell line, centrifuge at 12,000 rpm for 30 minutes and collect the supernatant. After quantification, the supernatant of each group was subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis ( sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE for short) was analyzed, and then the proteins on the sodium dodecyl sulfate polyacrylamide gel were transferred to polyvinylidene difluoride (PVDF for short) on, and then use drugs targeting melanocortin 1 receptor (MC1R), tyrosinase, tyrosinase-related protein 1 (TRP1) and more Antibodies against dopachrome tautomerase (DCT) were used to detect the protein content in the supernatants of each group, and the protein content of β-actin (β-actin) was used as the internal control group. (internal control), converted into the relative protein content of a protein relative to β-actin (β-actin). Among them, melanocortin receptor 1 (MC1R), after binding to α-melanocyte-stimulating hormone (α-MSH), causes intracellular cyclic adenosine monophosphate, The content of cAMP for short increases, thereby promoting the expression of proteins such as tyrosinase (tyrosinase), tyrosinase-related protein 1 (TRP1) and dopachrome tautomerase (DCT). Tyrosinase can convert DOPA in melanocytes is oxidized to form dopaquinone, which then forms dopachrome, and is finally converted into eumelanin; dopachrome undergoes intermutation of dopachrome The action of DCT can form 5,6-dihydroxyindole-2-carboxylic acid (DHICA) and then eumelanin, 5,6 -Dihydroxyindole-2-carboxylic acid (DHICA) can also form eumelanin through the action of tyrosinase-related protein 1 (TRP1), in other words, melanocortin receptor 1 (MC1R), Tyrosinase, tyrosinase-related protein 1 (TRP1) and dopachrome tautomerase (DCT) are important proteins in the message transmission pathway that promotes melanin production.

Please refer to Table 4 and Figure 5. In group D1, whether it is melanocortin receptor 1 (MC1R), tyrosinase (tyrosinase), tyrosinase-related protein 1 (TRP1) and The relative protein content of dopachrome tautomerase (DCT) has been significantly reduced, which can confirm that the whitening composition can indeed block the message transmission pathway that promotes melanin production, thereby effectively reducing melanin production.

In summary, the whitening composition of the present invention, composed of the grass coral extract, the rosemary extract and the Andrographis paniculata extract, can not only effectively inhibit the activity of tyrosinase, but also block the activity of tyrosinase. The message transmission pathway related to melanin production can thereby reduce the production of melanin. Therefore, the whitening composition can be used to prepare a whitening preparation for administration to a desired individual to inhibit the production of melanin in the melanocytes of the desired individual. , which is the effect of the present invention.

Furthermore, the use of the whitening composition of the present invention can further achieve the effect of effectively inhibiting the production of melanin by using the aforementioned whitening composition.

Although the present invention has been disclosed using the above-mentioned preferred embodiments, they are not intended to limit the invention. Anyone skilled in the art can make various changes and modifications to the above-described embodiments without departing from the spirit and scope of the invention. The technical scope protected by the invention, therefore, the protection scope of the invention shall include all changes within the literal and equivalent scope described in the appended patent application scope.

﹝This invention﹞ S1: Preparation steps of grass coral extract S11: Sample provision sub-step S12: Extraction sub-step S13: Concentration sub-step S2: Rosemary extract preparation steps S21: Sample provision sub-step S22: Extraction sub-step S23: Concentration sub-step S3: Preparation steps of Andrographis paniculata extract S31: Sample provision sub-step S32: Extraction sub-step S33: Concentration sub-step S4: Modulation step

[Figure 1] A flow chart of a method for manufacturing a whitening composition according to an embodiment of the present invention. [Picture 2] In test (A), the histogram of the relative tyrosinase activity of the samples to be tested in groups A0 to A8 (*: P <0.05, compared with group A0; #: P <0.01, Compared with group A2; @: P <0.05, compared with group A3; $: P <0.05, compared with group A4; %: P <0.05, compared with group A5; &: P < 0.05, compared with group A7). [Figure 3] In test (B), the bar graph of the relative tyrosinase activity of the samples to be tested in groups B0 to B5 (*: P <0.05, compared with group B0; #: P <0.01, Compared with group B2; $: P <0.05, compared with group B5). [Picture 4] In the test (C), the histogram of the relative intracellular melanin content of the samples to be tested in groups C0 to C3. [Figure 5] In test (D), the relative protein content histogram of the samples to be tested in groups D0 to D1.

S1: Preparation steps of grass coral extract

S11: Sample provision sub-step

S12: Extraction sub-step

S13: Concentration sub-step

S2: Rosemary extract preparation steps

S21: Sample provision sub-step

S22: Extraction sub-step

S23: Concentration sub-step

S3: Preparation steps of Andrographis paniculata extract

S31: Sample provision sub-step

S32: Extraction sub-step

S33: Concentration sub-step

S4: Modulation step

Claims (8)

A whitening composition, comprising: a herbaceous coral extract; a rosemary extract; and an andrographis paniculata extract, wherein the herbaceous coral extract, the rosemary extract and the andrographis paniculata extract are based on a phosphate Buffered physiological saline solution was used as the extraction agent to extract a Coral Coral sample, a Rosemary sample and an Andrographis paniculata sample at a pressure between 1 and 1.2 Kgf/cm ² and a temperature between 121 and 135°C. obtained. The whitening composition of claim 1, wherein the phosphate buffered physiological saline solution contains sodium chloride with a concentration between 0.12 and 0.14 M and sodium dihydrogen phosphate with a concentration between 0.008 and 0.012 M, and the Phosphate buffered saline solution has a neutral pH. The whitening composition of claim 1, wherein, after obtaining the Herba Coral extract, the Rosemary extract and the Andrographis Paniculata extract respectively, the Herba Coral extract, the Rosemary extract and the Andrographis paniculata are mixed. extract to form the whitening composition. The whitening composition as claimed in any one of claims 1 to 3, wherein the whitening composition includes 15 to 55% by weight of grass coral extract, 30 to 55% of rosemary extract and 15 to 15% by weight. 55% Andrographis paniculata extract. Such as the whitening composition of claim 4, wherein the whitening composition includes 33.3% of Coral grass extract, 50% of Rosemary extract and 16.7% of Andrographis paniculata extract by weight percentage. The use of a whitening composition is applied to the preparation of whitening preparations, wherein the whitening composition is the whitening composition described in any one of claims 1 to 5, and is administered to a desired individual to inhibit the The production of melanin in the melanocytes of the desired individual. The use of the whitening composition of claim 6, wherein the whitening composition is administered to the desired individual in the form of smearing or spraying. The use of the whitening composition of claim 6, wherein the whitening composition is administered to the desired individual at a dose of 200 to 500 μg/cm ² .

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