Pharmaceutical composition for preventing and treating chloasma

Abstract

The invention discloses a composition for preventing and / or treating chloasma, which comprises ligusticum wallichii total alkaloids and leonurus total alkaloids in a mass ratio of (1: 4)-(5: 1). According to the pharmaceutical composition for preventing and treating chloasma, disclosed by the invention, the rhizoma chuanxiong alkaloid and the herba leonuri alkaloid are prepared according to a specific proportion to generate a synergistic effect in the aspect of inhibiting melanocyte proliferation. Animal experiments confirm that the composition can achieve whitening and freckle removing effects equivalent to those of hydroquinone cream, and has clinical popularization value.

Description

Technical Field

[0001] The invention belongs to the field of pharmaceutical preparations, and in particular relates to a pharmaceutical combination for preventing and treating chloasma. Background Art

[0002] Melasma is an acquired pigmentation characterized by bilateral irregular brown spots and plaques that appear on the sun-exposed areas of the face and rarely on the forearms. The etiology of melasma is mainly related to exposure to ultraviolet (UV) radiation and visible light, familial susceptibility, pregnancy and the use of exogenous hormones, which cause tyrosinase activity and increase melanin production. Studies have shown that the occurrence of melasma also involves basement membrane zone destruction, photoaging, oxidative stress, keratinocytes, mast cells, abnormal gene regulation, inflammatory response, neovascularization, and skin barrier damage. The purpose of treating melasma patients is to reduce facial spots, reduce the diameter of spots, and avoid recurrence. There are many methods for treating melasma in Western medicine, mainly including topical medications, chemical peels, laser treatments, and oral medications. However, the treatment measures have obvious disadvantages, and there are cases where the drugs have large side effects or poor efficacy.

[0003] Chloasma is classified as "facial dust" and "liver spots" in traditional Chinese medicine, and the advantages of traditional Chinese medicine in the treatment of chloasma are attracting more and more attention. Wang Yongheng et al. used DPPH free radical scavenging rate and mushroom tyrosinase inhibition rate as evaluation indicators in "Study on Dual-target Activity of Chuanxiong Extract for Whitening", screened the activity of Chuanxiong extracts extracted with different solvents and different polar parts, and found that the ethanol water extract of Chuanxiong can inhibit the activity of intracellular tyrosinase, thereby reducing the production of melanin. At the same time, it has an antioxidant effect, can scavenge free radicals, and reduce the oxidative stress response under ultraviolet stimulation, thereby achieving the effect of removing spots and whitening. Miao Chunping et al. pointed out in "A Brief Discussion on the Clinical Application of Motherwort in Dermatology" that motherwort can improve the blood circulation of the uterus and ovaries; regulate sex hormones and promote the secretion level of sex hormones, and have a very good effect in the treatment of chloasma.

[0004] However, there are no reports on the anti-freckle active ingredients of Chuanxiong and Leonurus japonicus, or the combined application of active ingredients in the treatment of chloasma. Summary of the invention

[0005] To solve the above problems, the present invention provides a composition for preventing and / or treating chloasma, which comprises total alkaloids of chuanxiong and total alkaloids of leonurus japonicus, and the mass ratio of total alkaloids of chuanxiong and total alkaloids of leonurus japonicus is between 1:4 and 5:1.

[0006] Furthermore, the mass ratio of the total alkaloids of Rhizoma Chuanxiong and the total alkaloids of Herba Leonuri is 5:1, 4:1, 3:1 or 1:3.

[0007] Furthermore, the external preparation is prepared from total alkaloids of Rhizoma Chuanxiong and total alkaloids of Herba Leonuri as active ingredients and auxiliary materials acceptable to medicines.

[0008] Furthermore, the external preparation is a solution, a spray, an aerosol, an ointment or an emulsion.

[0009] Furthermore, the total alkaloids of Chuanxiong are extracted by adding 80% ethanol to Chuanxiong, the extract is loaded on AB-8 macroporous adsorption resin, washed with water to remove impurities, and then the eluate after elution with 85% ethanol is dried to obtain the extract.

[0010] Furthermore, the total alkaloids of motherwort are extracted by adding 90% ethanol to motherwort, the extract is adjusted to pH 1-2, loaded on a strong acid cationic resin, eluted with water to neutrality, then eluted with sodium chloride solution, the eluate is collected and concentrated, and anhydrous ethanol is added to extract the concentrated extract.

[0011] The present invention also provides a method for preparing the aforementioned composition, which comprises the following steps:

[0012] The total alkaloids of Rhizoma Chuanxiong and Herba Leonuri are weighed according to the proportion, mixed, and added with pharmaceutically acceptable auxiliary materials to obtain the product.

[0013] The present invention also provides a use of the aforementioned composition in preparing a medicine for preventing and / or treating chloasma.

[0014] Furthermore, the drug has the effect of inhibiting melanocyte proliferation, tyrosinase activity, melanin production and / or oxidative stress.

[0015] The pharmaceutical composition for preventing and treating chloasma of the present invention produces a synergistic effect in inhibiting the proliferation of melanocytes by combining ligusticum chuanxiong alkaloids and leonurus alkaloids in a specific ratio. Animal experiments have shown that the composition of the present invention can achieve a whitening and freckle-removing effect comparable to that of hydroquinone cream, and has clinical promotion value.

[0016] Obviously, according to the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modification, replacement or change may be made.

[0017] The above contents of the present invention are further described in detail below through specific implementation methods in the form of embodiments. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1The combined use index (CI) of the total alkaloids of Chuanxiong and Leonurus japonicus in different proportions;

[0019] Figure 2 When the ratio of total alkaloids of Rhizoma Chuanxiong and total alkaloids of Leonurus japonicus = 1:2, the CI index and inhibition rate change;

[0020] Figure 3 Effects of the combination of Leonurus japonicus and Chuanxiong alkaloids on the auricle histopathology of chloasma mice;

[0021] Figure 4 Effect of the combination of Leonurus japonicus and Ligusticum chuanxiong alkaloids on the melanin content in the auricle of chloasma mice.

[0022] Figure 5 Effect of the combination of Leonurus japonicus and Ligusticum chuanxiong alkaloids on the distribution of melanin in the auricle of chloasma mice. DETAILED DESCRIPTION

[0023] The raw materials and equipment used in the specific embodiments of the present invention are all known products and can be obtained by purchasing commercial products. The preparation process of the solution, spray, aerosol, ointment or emulsion prepared from the composition of the present invention is mature and can be prepared by referring to the conventional method of the prior art.

[0024] Example 1 Preparation of the composition of the present invention

[0025] Take the total alkaloids of Rhizoma Chuanxiong and the total alkaloids of Herba Leonuri, mix them in a mass ratio of 5:1, add pharmaceutically acceptable excipients and mix well to obtain the product.

[0026] Example 2 Preparation of the composition of the present invention

[0027] Take the total alkaloids of Rhizoma Chuanxiong and the total alkaloids of Herba Leonuri, mix them in a mass ratio of 4:1, add pharmaceutically acceptable excipients and mix well to obtain the product.

[0028] Example 3 Preparation of the composition of the present invention

[0029] Take the total alkaloids of Rhizoma Chuanxiong and the total alkaloids of Herba Leonuri, mix them in a mass ratio of 3:1, add pharmaceutically acceptable excipients and mix well to obtain the product.

[0030] Example 4 Preparation of the composition of the present invention

[0031] Take the total alkaloids of Rhizoma Chuanxiong and the total alkaloids of Herba Leonuri, mix them in a mass ratio of 1:3, add pharmaceutically acceptable excipients and mix well to obtain the product.

[0032] The beneficial effects of the present invention are described below through test examples.

[0033] Experimental Example 1 Effect of the combination of effective components of Chuanxiong and Leonurus japonicus on melanoma cells

[0034] 1. Materials and Methods

[0035] 1.1 Trial Drugs

[0036] The total alkaloids of Ligusticum chuanxiong and Leonurus japonicus were provided by the Chemical Laboratory of Chengdu University of Traditional Chinese Medicine.

[0037] The preparation method of total alkaloids of Leonurus japonicus is as follows:

[0038] Take 5kg of dried medicinal materials of motherwort, extract twice with 90% ethanol in 9 times (v / w, mL / g) volume, 129min each time, combine the solvents, and concentrate to obtain the total alkaloid extract of motherwort. Adjust the pH of the motherwort extract to 1-2 with hydrochloric acid, filter, and then control the flow rate of the liquid to 4BV / h, pass through a strong acid cationic resin column (001×7 type) with a column diameter and column height ratio of 1:8 in the forward direction, wash with water to neutrality, select sodium chloride solution (3.0mol / L) as the eluent, collect 10 times the column volume of eluent (until the alkaloid reaction of the effluent is not positive), and concentrate to extract. Further use anhydrous ethanol for ultrasonic extraction until the extract alkaloid reaction is negative, concentrate the extract to prepare the total alkaloid of motherwort, and finally purify to obtain 40g of total alkaloid of motherwort.

[0039] The alkaloid reaction method used in preparing the total alkaloids of Leonurus japonicus can adopt conventional methods such as modified bismuth potassium iodide reaction or iodine-potassium iodide reaction. The present invention specifically adopts a modified Dragendorff reaction, that is, taking the sample to be tested and adding the modified Dragendorff reagent (bismuth nitrate and potassium iodide in an alkaline aqueous solution), if the alkaloid is present, an orange-red or brown precipitate will be formed, which is judged as positive, otherwise it is negative; iodine-potassium iodide reaction: after taking the sample to be tested and mixing it with the iodine solution and the potassium iodide solution, if a yellow precipitate appears, it is judged as positive, otherwise it is negative.

[0040] The preparation method of total alkaloids of Ligusticum chuanxiong is as follows:

[0041] Take 10kg of Chuanxiong medicinal materials, add 15 times the amount (v / w, mL / g) of 80% ethanol, and reflux extract for 2.5h. The total alkaloid extract of Chuanxiong was further purified by static adsorption separation. The resin used in the purification process was AB-8 macroporous adsorption resin. The specific purification process conditions of total alkaloids of Chuanxiong were as follows: the pH of the sample solution was 8.0, and the mass concentration of total alkaloids of Chuanxiong in the sample solution was 4.4799mg·mL -1 The sample volume was 8 BV. After removing impurities with 13 BV of deionized water, elution was performed with 14 BV of 85% ethanol by volume, and concentration was performed to obtain 120 g (extraction rate was 1.2%) of total alkaloids of Rhizoma Chuanxiong.

[0042] 1.2 Experimental Reagents

[0043] CCK-8 (Shanghai Shenggong Bioengineering Co., Ltd., batch number: K409FC0256), levodopa (Shanghai Yuanye Biotechnology Co., Ltd., batch number: J03HS173328), TritonX-100 (Shanghai Biyuntian Technology Co., Ltd., batch number: ST797), dimethyl sulfoxide (Chengdu Kelong Chemical Co., Ltd., batch number: 2022041502), 1640 culture medium (Gibco, batch number: 8123576), DMEM culture medium (Gibco, batch number: 8123576), fetal bovine serum (Hyclone, batch number: SF0007001), trypsin digestion solution (Solarbio, batch number: 242806001) penicillin-streptomycin solution (double antibody) (Biosharp, batch number: 20200617).

[0044] 2 Experimental methods

[0045] 2.1 Cell culture and passaging

[0046] Under sterile conditions, B16 mouse melanoma cells were inoculated in RPMI-1640 medium or DMEM medium containing 10% fetal bovine serum and 1% double antibody, and cultured in an incubator at 37°C, 5% CO2, and 98% relative humidity. When the cells grew to a confluence of 80% to 90%, they were digested with 0.25% trypsin (containing 0.05% EDTA), subcultured, and the logarithmic phase cells that had been subcultured for more than 3 generations after recovery were collected for subsequent experiments.

[0047] 2.2 Effects of the combination of Chuanxiong alkaloids and Leonurus japonicus alkaloids on B16 cell proliferation

[0048] According to the mass ratio of total alkaloids of Chuanxiong and total alkaloids of Leonurus japonicus of 1:0, 0:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 5:1, the drug-containing complete medium with a final total concentration of the two drugs added was 4, 2, 1.5, 1, 0.5, and 0.25 mg / mL. When the cells grew to 80% to 90% confluence, they were digested with 0.25% trypsin, collected and prepared into single cell suspension, and the cell density was adjusted to 5×10 with fresh culture medium. 4 / mL, inoculated into 96-well culture plates, 100μL per well, placed in an incubator and cultured overnight. After the cells adhered to the wall, the supernatant was discarded and replaced with a drug-containing complete medium with a final drug concentration of 4, 2, 1.5, 1, 0.5, and 0.25mg / mL. At the same time, a control group without drug addition and a blank group without drug addition or cells were set up, with 3 replicates in each group and 100μL per well. After 48h of culture, each well was washed 3 times with PBS and replaced with a fresh medium containing 10% CCK-8 solution. The wells were incubated in the dark for 3h and the absorbance (OD) value of each well was detected at a wavelength of 450nm. Cell inhibition rate = (test well OD450 nm - control well OD450nm) / (control well OD450 nm - blank well OD450nm).

[0049] 3 Statistical methods

[0050] GraphPad Preism software was used for data analysis. The quantitative data were expressed as mean ± standard deviation. One-way ANOVA was used, and P < 0.05 or P < 0.01 indicated that the difference between the two groups was statistically significant.

[0051] 4 Experimental results

[0052] The CCK-8 method is based on WST-8, a water-soluble tetrazolium salt, which can be reduced by dehydrogenases in cell mitochondria to a highly water-soluble yellow formazan product. The amount of formazan generated is proportional to the number of living cells. The microplate reader measures the light absorption value of the formazan product at a wavelength of 450nm, which can indirectly reflect the survival and growth of cells. The CCK8 results showed that total alkaloids from Chuanxiong and Leonurus japonicus can effectively inhibit the proliferation of B16 cells, and the half inhibition rate IC of total alkaloids from Chuanxiong and Leonurus japonicus on B16 cells was calculated. 50 They were 2.30±0.03mg / mL and 0.91±0.02mg / mL respectively.

[0053] The Chou-Talalay method, also known as the Median-Drug Effect Analysis or Combination Index Method, is a digital method for quantitatively calculating the combined effects of drugs. The CI index calculation formula is: Compusyn software is a software developed based on the Chou-Talalay method to calculate the combination index CI value. The CI value calculated by the software is used to draw the combination index diagram of each ratio ( Figure 1 ) and inhibition rate-combination index diagram ( Figure 2 ).

[0054] When the CI value is less than 1, it means that the combined use of the two drugs has produced a synergistic effect. From Figure 1 It can be seen that when the mass ratio of total alkaloids of Ligusticum chuanxiong Hort. to total alkaloids of Leonurus japonicus Houtt. is lower than 5:1 and higher than 1:4, a coordinated and synergistic effect is produced in inhibiting the proliferation of melanocytes. From Figure 2 It can be seen that as the coordination index CI value of the combination of total alkaloids of Ligusticum chuanxiong Hort. and total alkaloids of Leonurus japonicus Houtt. decreases, the inhibition rate of melanoma cells gradually increases.

[0055] From the above results, it can be seen that the compatibility of alkaloids of Ligusticum chuanxiong Hort. and alkaloids of Leonurus japonicus Houtt. can significantly inhibit the proliferation of melanocytes. When the mass percentage of total alkaloids of Ligusticum chuanxiong Hort. and total alkaloids of Leonurus japonicus Houtt. is between 1:4 and 5:1, specifically 5:1, 4:1, 3:1 or 1:3, the coordination index CI values of inhibiting melanocytes are all less than 1, and a synergistic effect is produced.

[0056] Experimental Example 2: Study on the effect of the combination of total alkaloids of Ligusticum chuanxiong Hort. and total alkaloids of Leonurus japonicus Houtt. on chloasma

[0057] 1 Experimental materials

[0058] 1.1 Animals

[0059] 48 SPF-grade C57BL / 6 mice, female, weighing (20±2) g, provided by Beijing Speifo Biotechnology Co., Ltd., with the experimental animal production license number SCXK (Chuan) 2020-030 and the use license number: SYXK (Chuan) 2020-0124. They were conventionally fed with standard mouse feed and freely ate and drank water in the same environment.

[0060] 1.2 Drugs and reagents

[0061] Total alkaloids of Ligusticum chuanxiong Hort. and total alkaloids of Leonurus japonicus Houtt. were both provided by the Chemical Laboratory of Chengdu University of Traditional Chinese Medicine; hydroquinone cream (Guangdong Renrenkang Pharmaceutical Co., Ltd., batch number: 20230203); progesterone injection (Zhejiang Xianju Pharmaceutical Co., Ltd., batch number: FB2229), sodium carboxymethylcellulose (Shanghai Yuanye Bio-Technology Co., Ltd., batch number: J07HS180136); hematoxylin staining solution (BASO, batch number: BA-4041); eosin staining solution (BASO, batch number: BA-4022); SOD and MDA kits (Nanjing Jiancheng, batch number of R0204: 20230628).

[0062] 1.3 Instruments

[0063] ME204E electronic balance (Mettler-Toledo Instrument Co., Ltd.); KQ5200DE CNC ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.); water purifier (Sichuan Youpu Ultrapure Technology Co., Ltd.); Allegra X-30R centrifuge (BeckMan Company), ultraviolet light therapy device (Shanghai Sigma High-Tech Co., Ltd.).

[0064] 2 Experimental methods

[0065] 2.1 Drug preparation method

[0066] Drug preparation: According to clinical medication, chuanxiong and motherwort alkaloids (1:2) were dissolved in 5% PEG-400 aqueous solution to prepare solutions with concentrations of 324 mg / mL, 162 mg / mL, and 81 mg / mL, and then the drugs were dissolved in 3% sodium carboxymethyl cellulose to prepare a 20 mL solution. The final concentrations were 16.2, 8.1, and 4.05 mg / mL. The low, medium, and high concentrations for mice were calculated to be 0.41 mg / (cm 2 ·d)、0.81mg / (cm 2 ·d)、1.62mg / (cm 2 ·d).

[0067] 2.2 Grouping, modeling and drug administration

[0068] The mice were randomly divided into a blank group, a model group, a positive control group (hydroquinone cream group), and a drug administration group, with 6 mice in each group.

[0069] The mouse auricle was modeled with ultraviolet light combined with progesterone. Except for the blank group, the other groups were irradiated with medium-wave ultraviolet light (UVB) with a wavelength of 280-320nm once a day for 30 minutes, and the light source was 20cm away from the mouse for 30 consecutive days. At the same time, except for the blank group, the other groups were intramuscularly injected with progesterone injection 20mg / (kg·d), while the blank group was intramuscularly injected with sterile injection water 2mL / (kg·d). All groups were injected into the thigh root of the mouse, once a day, alternating between the two legs, for 30 consecutive days. After the modeling, the blank group and the model group were smeared with blank matrix on the auricle, the positive drug group was smeared with hydroquinone cream, and the drug group was evenly smeared with alkaloid drugs.

[0070] 2.3 Specimen collection

[0071] After the last administration, the mice were fasted but not watered for 24 hours, and their weights were measured. Blood was collected from the eye sockets of the mice, and the blood was allowed to coagulate at room temperature. The blood was centrifuged at 4°C and 3500 r / min for 15 minutes, and the supernatant was collected and stored in a -80°C refrigerator for later use. The model tissues of the backs of the mice in each group were cut and fixed with 4% paraformaldehyde solution for later use.

[0072] 2.4 Detection indicators

[0073] 2.4.1 Detection of SOD and MDA content in mouse auricle tissue

[0074] The mouse auricle tissue was washed with ice-cold physiological saline, and the auricle tissue was accurately weighed and diluted to 10 mg / 100 μL. -1 The mixture was precooled to 4°C in a cryo-grinder and ground for 60 s × 6 times at 45 Hz, with an interval of 10 s between each grinding. The mixture was centrifuged for 15 min (4°C, 12000 r·min-1). The supernatant was separated and the SOD activity and MDA content of the mouse auricle tissue were detected according to the instructions of the kit.

[0075] 2.4.2 Observation of pathological changes in mouse auricle tissue and melanin distribution by H&E and Masson Fontana staining

[0076] HE staining: The auricle tissue fixed with 4% paraformaldehyde was routinely dehydrated, paraffin-embedded, sectioned, and HE-stained. The pathological morphological changes of the auricle tissue were observed under a microscope.

[0077] Masson Fontana staining: The fixed tissue was sliced, then placed in distilled water, and then Fontana ammonia silver solution was added, and the slices were stained in the dark for 12-24 hours. After the staining, the slices were washed with distilled water 5-6 times, each time for 1-2 minutes. Then, the slices were treated with Hypo solution for 1-5 minutes, followed by tap water for 3-5 minutes, dehydrated with 95% ethanol and anhydrous ethanol, transparentized with xylene, and sealed with neutral gum. Image-Pro Plus 6.0 was used to measure the integrated optical density (IOD) of tissue melanin.

[0078] 3 Statistical methods

[0079] SPSS26.0 statistical software was used for data analysis. The measurement data were expressed as mean ± standard deviation. One-way ANOVA was used, and P < 0.05 or P < 0.01 indicated that the difference between the two groups was statistically significant.

[0080] 4 Experimental results

[0081] 4.1 SOD activity and MDA content in mouse auricle

[0082] Compared with the blank group, the SOD activity in the auricle tissue of the mice in the model group was significantly downregulated (P<0.01), and the MDA level was significantly upregulated (P<0.01). Compared with the model group, the SOD activity in the hydroquinone cream group, the low, medium and high dose groups were significantly upregulated (P<0.01); compared with the model group, the MDA content in the hydroquinone cream group was significantly reduced (P<0.01), and the MDA content in the low, medium and high dose groups was significantly downregulated (P<0.05). This shows that the combination of alkaloids can effectively reduce the body's oxidative stress level. The specific experimental results are shown in Table 1.

[0083] Table 1 Effects of drug combination on SOD activity and MDA content in mouse auricle (n=8, )

[0084]

[0085] Note: Compared with the blank group, ** P<0.01; compared with the model group # P<0.05, ## P<0.01.

[0086] 4.2 Pathological changes of mouse auricle tissue

[0087] The auricle tissue of the blank group was basically normal. Compared with the blank group, the auricle of the mice in the model group, positive group, and drug-treated group showed no obvious pathological changes; compared with the model group, the auricle tissue of the mice in the positive group and drug-treated group showed no obvious changes. This shows that the alkaloid combination is safe and has no toxic side effects. For specific results, see Figure 3 .

[0088] 4.3 Changes in melanin content in mouse auricle tissue

[0089] Masson Fontana staining revealed that a small amount of melanin was distributed in the auricle tissue of the blank group mice; compared with the blank group, the distribution of melanin in the auricle tissue of the model group mice increased significantly, and the IOD value increased significantly, with statistical significance (P<0.01); compared with the model group, the distribution of melanin in the auricle of the positive group mice decreased significantly, and the IOD value decreased significantly, with statistical significance (P<0.01); compared with the model group, the distribution of melanin in the auricle of the low-dose mice showed a decreasing trend; compared with the model group, the distribution of melanin in the auricle of the medium- and high-dose groups mice decreased significantly, and the IOD value decreased significantly, with statistical significance (P<0.01). Figure 4 and Figure 5 This indicates that the alkaloid combination has the effect of significantly reducing melanin deposition, which is equivalent to the effect of hydroquinone cream and can effectively treat melasma.

[0090] Table 2 Melanin integrated optical density in mouse auricle tissue (n=6, )

[0091]

[0092] Note: Compared with the blank group, * P<0.01; compared with the model group # P<0.01.

[0093] In summary, the composition of the chuanxiong alkaloids and the motherwort alkaloids of the present invention produces a synergistic effect in inhibiting the proliferation of melanocytes. Under the strongest synergistic effect between the chuanxiong alkaloids and the motherwort alkaloids, the tyrosinase activity of melanocytes is inhibited, the melanin production is reduced, and the whitening and freckle removal effect comparable to that of the hydroquinone cream can be achieved.

Claims

1. A composition for preventing and / or treating chloasma, characterized in that: The composition includes total alkaloids of chuanxiong rhizome and total alkaloids of leonurus japonicus, and the mass ratio of total alkaloids of chuanxiong rhizome and total alkaloids of leonurus japonicus is between 1:4 and 5:

1.

2. The composition according to claim 1, characterized in that: The mass ratio of the total alkaloids of Rhizoma Chuanxiong and the total alkaloids of Herba Leonuri is 5:1, 4:1, 3:1 or 1:

3.

3. The composition according to claim 1 or 2, characterized in that: The external preparation is prepared from total alkaloids of Rhizoma Chuanxiong and total alkaloids of Herba Leonuri as active ingredients and auxiliary materials acceptable to medicines.

4. The composition according to claim 3, characterized in that: The external preparation is a solution, a spray, an aerosol, an ointment or an emulsion.

5. The composition according to any one of claims 1 to 3, characterized in that: The total alkaloids of Chuanxiong are extracted by adding 80% ethanol to Chuanxiong, the extract is loaded on AB-8 macroporous adsorption resin, washed with water to remove impurities, and then the eluate is eluted with 85% ethanol, and the extract is dried.

6. The composition according to any one of claims 1 to 3, characterized in that: The total alkaloids of motherwort are extracted by adding 90% ethanol to motherwort, adjusting the pH value of the extract to 1-2, loading it on a strong acidic cationic resin column, eluting it with water until it is neutral, then eluting it with a sodium chloride solution, collecting and concentrating the eluate, and adding anhydrous ethanol to extract the concentrated extract.

7. A method for preparing the composition according to any one of claims 1 to 6, characterized in that: It includes the following steps: The total alkaloids of Rhizoma Chuanxiong and Herba Leonuri are weighed according to the proportion, mixed, and added with pharmaceutically acceptable auxiliary materials to obtain the product.

8. Use of the composition according to any one of claims 1 to 6 in the preparation of a medicament for preventing and / or treating chloasma.

9. The use according to claim 8, characterized in that: The drug has the effect of inhibiting the proliferation of melanocytes.

10. The use according to claim 8, characterized in that: The drug has the effect of inhibiting tyrosinase activity, melanin production and / or oxidative stress.

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