Application of desert algae extract in preparation of product for treating vitiligo
Through the study of desert algae extract, it was found that desert algae extract can effectively improve tyrosinase activity, promote melanin accumulation, solve the problem of insufficient existing technology in the treatment of vitiligo, provide new treatment ideas, and has wide application prospects due to its natural and safe nature.
Patent Information
- Application Number
- CN202411835058.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-13
- Publication Date
- 2025-03-14
AI Technical Summary
The prior art has not yet effectively utilized the application of desert algae extract in the treatment of vitiligo, and there is a lack of relevant reports.
Through research, it was found that desert algae water extract, desert algae extract, desert algae water extract ethyl acetate extract and desert algae ethyl acetate extract can effectively improve tyrosinase activity and promote melanin accumulation, thus becoming candidate drugs for the treatment of vitiligo.
These desert algae extracts showed that at lower concentrations, they could increase tyrosinase activity and promote melanin accumulation, providing new ideas for the treatment of vitiligo, and because desert algae is a natural and edible algae, they are highly safe.
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Figure CN119607031A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of biomedicine, and in particular relates to an application of a desert algae extract in preparing a product for treating vitiligo. Background Art
[0002] Vitiligo is an autoimmune disease and a depigmented skin disease, which is mainly manifested by white spots and gray / white hair caused by the selective reduction or disappearance of melanocytes in the skin, mucous membranes and hair. Research results show that vitiligo is a localized or generalized depigmented lesion caused by the progressive reduction or disappearance of melanin granule production due to the reduction or disappearance of tyrosinase activity in melanocytes in the skin and hair follicles.
[0003] Desert algae has biological activities such as enhancing immunity, regulating blood pressure, lowering blood lipids, anti-tumor, anti-infection, and anti-inflammation. However, there are no reports on the use of desert algae in the treatment of vitiligo. Summary of the invention
[0004] In order to solve the above technical problems, the present invention provides an application of a desert algae extract in the preparation of a product for treating vitiligo. The present invention has been found through research that desert algae water extract, desert algae alcohol extract, desert algae water-extracted ethyl acetate extract, and desert algae alcohol-extracted ethyl acetate extract can effectively improve tyrosinase activity in vitro and effectively promote melanin accumulation. Therefore, these desert algae extracts can become candidate drugs for treating vitiligo, including non-segmental vitiligo, segmental vitiligo, and uncertain vitiligo, and can also be made into functional products such as health products and cosmetics. The desert algae used in the present invention is natural, edible algae with high safety. The present invention has opened up new ideas for the treatment of vitiligo.
[0005] In order to achieve the above object, the present invention adopts the following technical solutions:
[0006] The invention provides an application of a desert algae extract in preparing a product for treating vitiligo.
[0007] Furthermore, the products for treating vitiligo include medicines for treating vitiligo, health products for treating vitiligo, and cosmetics for treating vitiligo.
[0008] Furthermore, the vitiligo includes non-segmental vitiligo, segmental vitiligo, and indeterminate vitiligo.
[0009] Furthermore, the desert algae extract includes desert algae water extract, desert algae alcohol extract, desert algae water-extracted ethyl acetate extract, and desert algae alcohol-extracted ethyl acetate extract.
[0010] The present invention also provides a product for treating vitiligo, which is a medicine for treating vitiligo, a health product for treating vitiligo, or a cosmetic for treating vitiligo, and the product contains the desert algae water extract, desert algae alcohol extract, desert algae water-extracted ethyl acetate extract, or desert algae alcohol-extracted ethyl acetate extract.
[0011] Preferably, the product also includes other auxiliary materials.
[0012] Further preferably, the excipient is any one or more of Qubai Babuqi tablets, Baishi pills, Vitiligo capsules, Qubai tincture, anthelmintic Vernonia injection, 8-methoxypsoralen, L-phenylalanine, Kailin, compound Kali cumin tincture, glucocorticoids, calcineurin inhibitors, and vitamin D3 derivatives.
[0013] Further preferably, the auxiliary material is any one or more of anthelmintic Vernonia, Psoralea corylifolia, Nigella sativa seeds, cloves, fennel, Anache, Elilesia salsa, Wedelia officinalis or their extracts.
[0014] Furthermore, the product also includes a pharmaceutically and cosmetically acceptable carrier, which is any one or more of a solubilizer, a cosolvent, an emulsifier, a colorant, a binder, a disintegrant, a filler, a lubricant, a wetting agent, an osmotic pressure regulator, a stabilizer, a glidant, a flavoring agent, a preservative, a suspending agent, a coating material, a fragrance, an anti-adhesive, an integrator, a penetration enhancer, a pH regulator, a buffer, a plasticizer, a surfactant, a thickener, a inclusion agent, a humectant, an absorbent, a diluent, a flocculant and a deflocculant, a filter aid, a release retardant, a polymer skeleton material and a film-forming material.
[0015] Furthermore, the dosage form of the product is any one of a solution, aromatic water, glycerin, tincture, spirit, mixture, emulsion, suspension, syrup, injection, powder, granule, capsule, tablet, pill, film, ointment, gel, liniment, plaster, paste, patch, sustained-release preparation, controlled-release preparation, microcapsule preparation, microsphere preparation, nano preparation, liposome, ointment, decoction, lotion, pill, aerosol, spray, powder aerosol, slurry, and extract.
[0016] Compared with the prior art, the present invention has the following technical effects:
[0017] (1) The present invention found through in vitro experiments that different extracts of desert algae can increase tyrosinase activity and promote melanoma accumulation in melanoma B16 cells at relatively low concentrations.
[0018] (2) The present invention finds that desert algae extract can be a candidate drug for treating vitiligo, including non-segmental vitiligo, segmental vitiligo and indeterminate vitiligo, and can also be made into functional products such as health care products and cosmetics.
[0019] (3) The desert algae used in the present invention are natural, edible algae and highly safe.
[0020] (4) The present invention opens up a new idea for the treatment of vitiligo. BRIEF DESCRIPTION OF THE DRAWINGS
[0021] Figure 1 The effect of each sample of a series of concentrations in Example 5 of the present invention on B16 cells for 24 hours;
[0022] Figure 2 The effects of each sample of a series of concentrations in Example 5 of the present invention on B16 cells for 48 hours;
[0023] Figure 3 The effects of the samples of the series of concentrations in Example 6 of the present invention on the tyrosinase activity of B16 cells;
[0024] Figure 4 The figure shows the effect of the samples of the series concentrations in Example 7 of the present invention on the melanin content of B16 cells. DETAILED DESCRIPTION
[0025] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention. Without departing from the spirit and essence of the present invention, modifications or substitutions made to the method, steps or conditions of the present invention are within the scope of the present invention. The reagents and instruments used in the following examples can all be obtained commercially, and the methods used in the examples are consistent with the conventional methods unless otherwise specified.
[0026] The technical solution of the present invention is further described in detail below in conjunction with embodiments.
[0027] The following examples are experiments for the preparation of different extracts of desert algae and the verification of the effectiveness of the extracts of desert algae. The experimental results were tested by statistical methods. All experiments were repeated at least 3 times independently, and the results were expressed as mean values and standard deviations, using one-way analysis of variance. All statistical analyses used P<0.05 as the test standard for significant statistical differences, and the analysis software was SPSS22.0 and GraphPad Prism5.
[0028] Example 1 Preparation of Desert Algae Water Extract
[0029] The instruments include centrifuge and oven.
[0030] Preparation method: 50g of Chlorella sorokinensis mud is crushed with an appropriate amount of water, the solid-liquid ratio is 1:5-1:15, centrifuged, the supernatant is collected, and then an appropriate amount of water is added, fully stirred, centrifuged, the supernatant is combined, the water extraction step is repeated, the supernatants are combined three times, concentrated, and dried to obtain the desert algae water extract.
[0031] Example 2 Preparation of desert algae ethanol extract
[0032] The reagent is ethanol.
[0033] The instruments include rotary evaporator, freeze dryer and Soxhlet extractor.
[0034] Preparation method: 50g of protein core chlorella mud is crushed with an appropriate amount of water, the solid-liquid ratio is 1:5-1:15, concentrated by rotary evaporation at 50°C, and freeze-dried. Weigh 5g of algae freeze-dried powder, soak it in 300mL of anhydrous ethanol overnight, extract it with Soxhlet (80-120°C), collect the extract, repeat the above extraction steps three times, combine the extracts, concentrate them, and dry them naturally to obtain the desert algae ethanol extract.
[0035] Example 3 Preparation of ethyl acetate extract from desert algae
[0036] The reagents include anhydrous ethanol and ethyl acetate.
[0037] The instruments include rotary evaporator, freeze dryer, Soxhlet extractor, and separatory funnel.
[0038] Preparation method: 50g of Scenedesmus mud is crushed with an appropriate amount of water, the solid-liquid ratio is 1:5-1:15, concentrated by rotary evaporation at 50℃, and freeze-dried. Weigh 5g of algae freeze-dried powder, add 300mL of anhydrous ethanol to soak overnight, Soxhlet extraction (80-120℃), collect the extract, repeat the above extraction steps three times, combine the extracts, concentrate, dry naturally, and obtain ethanol extract. Take 1g of ethanol extract, add 100mL of distilled water, obtain 100mL of alcohol-extracted water-soluble matter, take 50mL of alcohol-extracted water-soluble matter, add 50mL of ethyl acetate, extract, repeat the above extraction steps three times, combine the extracts, concentrate, and dry to obtain the desert algae alcohol-extracted ethyl acetate extract.
[0039] Example 4 Preparation of ethyl acetate extract from desert algae
[0040] The reagent is ethyl acetate.
[0041] The instruments include rotary evaporator, freeze dryer and separatory funnel.
[0042] Preparation method: 50g of Chlamydomonas mud is crushed with an appropriate amount of water, the solid-liquid ratio is 1:5-1:15, concentrated by rotary evaporation at 50°C, and freeze-dried. Weigh 1g of lyophilized algae powder and add 100mL of distilled water, soak overnight to obtain a water extract, take 50mL of the water extract and add 50mL of ethyl acetate to extract, repeat the above extraction steps three times, combine the extracts, concentrate, and dry to obtain the desert algae water-extracted ethyl acetate extract.
[0043] Example 5 Cytotoxicity test of different extracts of desert algae
[0044] 1. Materials and Reagents
[0045] The test samples are the ethanol extract of the desert algae in Example 2, the ethyl acetate extract of the desert algae extracted with alcohol in Example 3, and the ethyl acetate extract of the desert algae extracted with water in Example 4.
[0046] The reference substance was 8-methoxypsoralen (8-MOP).
[0047] The tumor cell line used for testing was melanoma B16 cell.
[0048] The reagents mainly include culture medium DMEM high glucose, fetal bovine serum, trypsin and EDTA digestion solution, MTT, and DMSO (dimethyl sulfoxide).
[0049] The test instruments include an optical inverted microscope, a centrifuge, a refrigerator, a culture bottle, a 96-well culture plate, an ultra-clean workbench, a CO2 incubator, an ELISA reader, and a high-pressure homogenizer.
[0050] 2. Experimental Methods
[0051] Melanoma B16 cells in the logarithmic growth phase were digested and seeded into a 96-well culture plate. 100 μl of cell suspension was added to each well, and the plate was placed in a 37° C., 5% CO 2 cell culture incubator for continued culture. After the cells adhered, the supernatant was discarded, and 200 μl of each sample solution with a series of concentrations (6.25 μg / ml, 2.5 μg / ml, 25 μg / ml, 50 μg / ml, 100 μg / ml, 200 μg / ml, 400 μg / ml, 800 μg / ml) was added as the sample group. The blank control group only added complete culture medium, and the positive control group added 8-MOP with a concentration of 10.81 μg / ml. After 48 hours of co-culture, 20 μl of MTT solution (with a concentration of 5 mg / ml) was added to each well, and the cells were placed in a cell culture incubator at 37°C and 5% CO2 for further culture for 4 hours. The culture plate was taken out, the supernatant was carefully discarded, 150 μl of dimethyl sulfoxide (DMSO) was added to each well, and the cells were oscillated on an oscillator for 5-10 minutes. The absorbance (optical density, OD) value was measured at 490 nm using an enzyme-labeled detector. The experiment should be repeated three times and the survival rate of B16 cells should be calculated using the formula.
[0052]
[0053] 3. Results
[0054] The results of the cell inhibition rate experiment are shown in Table 1. The effects of desert algae extract on B16 cells for 24 hours are shown in Table 1. Figure 1 As shown, the effect of 48h on B16 cells is as follows Figure 2 shown.
[0055] Table 1 IC50 of desert algae extract
[0056] sample IC50(24h) IC50(48h) Anhydrous ethanol 907.2 μg / mL 2841 μg / mL Water Extraction Ethyl Acetate Extraction 1481 μg / mL 4333.65μg / mL Anhydrous ethanol ethyl acetate extract 1152 μg / mL 804.2 μg / mL
[0057] Depend on Figure 1-2 It can be seen that the three desert algae extracts have no significant difference in cell proliferation. As shown in Table 1, the IC50 of different extracts in the cell inhibition rate experiment is above 800μg / mL. Therefore, the three desert algae extracts have no cytotoxicity.
[0058] Example 6 Effects of different extracts of desert algae on tyrosinase activity
[0059] 1. Materials and Reagents
[0060] The test samples are the ethanol extract of the desert algae in Example 2, the ethyl acetate extract of the desert algae extracted with alcohol in Example 3, and the ethyl acetate extract of the desert algae extracted with water in Example 4.
[0061] The reference substance was 8-methoxypsoralen (8-MOP).
[0062] The tumor cell line used for testing was melanoma B16 cell.
[0063] The reagents mainly include L-DOPA, DMEM high glucose culture medium, fetal bovine serum, trypsin and EDTA digestion solution, and RIPA cell lysis solution.
[0064] The test instruments include an optical inverted microscope, a centrifuge, a refrigerator, a culture bottle, a 6-well culture plate, an ultra-clean workbench, a CO2 incubator, an ELISA reader, and a high-pressure homogenizer.
[0065] 2. Experimental Methods
[0066] Take cells in the logarithmic growth phase and inoculate them in 6-well plates. After the cells are completely attached, add drugs, add different extracts of desert algae at a series of concentrations (12.5μg / ml, 25μg / ml, 50μg / ml, 100μg / ml, 200μg / ml), the blank control group only adds complete culture medium, and the positive control group adds 8-MOP at a concentration of 10.81μg / ml. After 48h, collect cells, lyse them, and determine protein content by Brandford method. Adjust the sample protein content to a uniform concentration, and after adding L-DOPA to each reaction system, measure the absorbance (Optical density, OD) value at 475nm.
[0067] 3. Results
[0068] The results are as follows Figure 3 shown.
[0069] Depend on Figure 3 It can be seen that there are extremely significant differences between high concentration of alcohol extract, high concentration of water-extracted ethyl acetate extract, low and high concentration of alcohol-extracted ethyl acetate extract (***P<0.001). It can be seen that the three desert algae extracts have a significant promoting effect on tyrosinase activity, and the high-quality concentration extract has a significant promoting effect on melanin synthesis.
[0070] Example 7 Effects of different extracts of desert algae on melanin content
[0071] 1. Materials and Reagents
[0072] The test samples are the ethanol extract of the desert algae in Example 2, the ethyl acetate extract of the desert algae extracted with alcohol in Example 3, and the ethyl acetate extract of the desert algae extracted with water in Example 4.
[0073] The reference substance was 8-methoxypsoralen (8-MOP).
[0074] The tumor cell line used for testing was melanoma B16 cell.
[0075] The reagents mainly include culture medium DMEM high glucose, fetal bovine serum, trypsin and EDTA digestion solution, and RIPA cell lysis solution.
[0076] The test instruments include an optical inverted microscope, a centrifuge, a refrigerator, a culture bottle, a 6-well culture plate, an ultra-clean workbench, a CO2 incubator, an ELISA reader, and a high-pressure homogenizer.
[0077] 2. Experimental Methods
[0078] Cells in the logarithmic growth phase were inoculated in 6-well plates. After the cells were completely attached to the wall, different extracts of desert algae were added with a series of concentrations (12.5μg / ml, 25μg / ml, 50μg / ml, 100μg / ml, 200μg / ml). The blank control group was only added with complete culture medium, and the positive control group was added with 8-MOP at a concentration of 10.81μg / ml. After 48 hours, the cells were collected and cell lysate was added, and the absorbance (Optical density, OD) value was measured at 470nm.
[0079] 3. Results
[0080] The results are as follows Figure 4 shown.
[0081] Depend on Figure 4 It can be seen that the melanin content of the anhydrous ethanol extracts of various concentrations is generally higher than that of other extracts. Except for the positive control, the melanin content of 200ug / ml anhydrous ethanol is the highest. The melanin content of the water-extracted ethyl acetate extract is higher than that of the alcohol-extracted ethyl acetate extract. The content of the alcohol-extracted ethyl acetate extract is less but still different. This shows that different extracts have a promoting effect on melanin synthesis.
[0082] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.
Claims
1. Application of desert algae extract in the preparation of products for treating vitiligo.
2. The use according to claim 1, characterized in that: The products for treating vitiligo include medicines for treating vitiligo, health products for treating vitiligo, and cosmetics for treating vitiligo.
3. The use according to claim 2, characterized in that: The vitiligo includes non-segmental vitiligo, segmental vitiligo, and indeterminate vitiligo.
4. The use according to claim 3, characterized in that: The desert algae extract includes desert algae water extract, desert algae alcohol extract, desert algae water-extracted ethyl acetate extract, and desert algae alcohol-extracted ethyl acetate extract.
5. A product for treating vitiligo, characterized in that: The product is a medicine for treating vitiligo, a health product for treating vitiligo or a cosmetic for treating vitiligo, and the product contains the desert algae water extract, desert algae alcohol extract, desert algae water extract ethyl acetate extract or desert algae alcohol extract ethyl acetate extract as described in claim 4.
6. The product according to claim 5, characterized in that The product also includes other excipients, which are any one or more of the following: anti-leukoplakia Babuqi tablets, Baishi pills, vitiligo capsules, anti-leukoplakia tincture, anthelmintic Vernonia injection, 8-methoxypsoralen, L-phenylalanine, kalin, compound kali cumin tincture, glucocorticoids, calcineurin inhibitors, and vitamin D3 derivatives.
7. The product according to claim 6, characterized in that The auxiliary materials are any one or more of anthelmintic Vernonia, Psoralea corylifolia, Nigella sativa seeds, cloves, fennel, Anache, Elilesia salsa, Wedelia officinalis or extracts thereof.
8. The product according to claim 5, characterized in that The product also includes pharmaceutically and cosmetically acceptable carriers, which are any one or more of solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrities, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants and deflocculating agents, filter aids, release retardants, polymer skeleton materials and film-forming materials.
9. The product according to claim 5, characterized in that The dosage form of the product is any one of solution, aromatic water, glycerin, tincture, spirit, mixture, emulsion, suspension, syrup, injection, powder, granule, capsule, tablet, pill, film, ointment, gel, liniment, plaster, paste, patch, sustained-release preparation, controlled-release preparation, microcapsule preparation, microsphere preparation, nano preparation, liposome, ointment, decoction, lotion, pill, aerosol, spray, powder aerosol, slurry and extract.