Application of mulberry extract in preparation of product for preventing and / or treating acne

Abstract

The invention relates to application of a mulberry extract in preparation of a product for preventing and / or treating acne. The mulberry extract comprises alkaloid with the weight content being 3% or above, and / or polysaccharide with the weight content being not higher than 70%, and / or flavone with the weight content being not higher than 10%, and / or amino acid with the weight content being not higher than 50%, and / or other components. The mulberry extract is good in acne treatment effect and small in side effect.

Description

Technical Field

[0001] The invention relates to the field of acne treatment, and in particular to application of a mulberry extract in preparing a product for preventing and / or treating acne. Background Art

[0002] Acne is an inflammatory disease of the pilosebaceous gland unit of the skin, mainly affecting the face and trunk, and mainly manifested as polymorphic inflammatory skin lesions such as comedones, papules, pustules, nodules, cysts, etc. Many factors such as excessive secretion of sebaceous glands, abnormal keratinization of the hair follicle duct opening, excessive proliferation of Propionibacterium acnes around the sebaceous gland tissue and the inflammatory response caused by it can lead to the occurrence of acne. Acne can cause permanent physical scars and have a negative impact on people's quality of life and self-image.

[0003] Currently, acne can be treated by topical administration (such as retinoids (retinoic acid, adapalene, etc.), antibiotics (minocycline, erythromycin, etc.), benzoyl peroxide, azelaic acid and salicylic acid, etc.) or systemic administration (such as oral antibiotics (ampicillin, azithromycin, doxycycline, etc.), hormone therapy (oral contraceptives COC or spironolactone), isotretinoin, etc.). The above topical or systemic therapies are accompanied by mild or severe adverse reactions, such as erythema, dryness, irritation, dermatitis, drug resistance, irregular menstruation, hyperkalemia, etc., which affect the efficacy of the drug or reduce the patient's compliance.

[0004] Therefore, it is of great significance to study drugs with fewer side effects and better acne treatment effects. Summary of the invention

[0005] In order to overcome the disadvantages of adverse reactions caused by the prior art for treating acne, the present invention provides a new use of mulberry extract.

[0006] The new use of the mulberry extract includes: use of the mulberry extract in preparing products for preventing and / or treating acne, and / or a method for preventing and / or treating acne using the mulberry extract.

[0007] To achieve the above object, the present invention is implemented through the following technical solutions:

[0008] Taking the weight percentage of the sum of the components of the mulberry extract as 100%, the mulberry extract contains alkaloids with a weight content of more than 3% (optionally containing alkaloids with a weight content of 3-99%, further optionally containing alkaloids with a weight content of 15-99%, further preferably, containing alkaloids with a weight content of 30-99%, further optionally containing alkaloids with a weight content of 40-99%, further optionally containing alkaloids with a weight content of 50-99%, further optionally containing alkaloids with a weight content of 60-99%),

[0009] and / or comprising a polysaccharide content of not more than 70% by weight (optionally comprising a polysaccharide content of 0.2-70% by weight, further optionally comprising a polysaccharide content of 0.2-50% by weight, further optionally comprising a polysaccharide content of 0.2-35% by weight, further optionally comprising a polysaccharide content of 0.2-25% by weight, further optionally comprising a polysaccharide content of 0.2-23% by weight, further optionally comprising a polysaccharide content of 20-25% by weight),

[0010] and / or contains flavonoids having a weight content of not more than 10% (optionally containing flavonoids having a weight content of 0.05-5%, further optionally containing flavonoids having a weight content of 0-2%, further optionally containing flavonoids having a weight content of 0.05-2%, further optionally containing flavonoids having a weight content of 0.5-1.5%, further optionally containing flavonoids having a weight content of 0-1%, further optionally containing flavonoids having a weight content of 0.05-1%),

[0011] and / or contains no more than 50% amino acids by weight (optionally contains 0-30% amino acids by weight, further optionally contains 0-25% amino acids by weight, further optionally contains 0-20% amino acids by weight, further optionally contains 0-5% amino acids by weight, further optionally contains 3-25% amino acids by weight or further optionally contains 5-20% amino acids by weight),

[0012] and / or other components (the weight content is optionally 0-25%, further optionally 0-20%, further optionally 0-15%, further optionally 0-11%, further optionally 2-20%, further optionally 4-8%).

[0013] Clinically, acne often occurs in areas rich in sebaceous glands, such as the face, neck, chest and / or back. According to the severity, acne is graded into mild blackheads and whiteheads, moderate second-level skin papules, moderate third-level skin inflammation aggravation with pustules, and severe fourth-level severe damage to the sebaceous gland structure, nodules and cysts in the dermis and epidermis, and eventually scars accompanied by pigmentation. The global acne grading system (GAGS) is a scoring method commonly used in clinical practice to assess the severity of acne patients. It is calculated based on the location and severity of acne and can reflect the number and severity of acne.

[0014] Optionally, the acne is selected from the group consisting of whiteheads, blackheads, papular acne, pustular acne, cystic acne, nodular acne, or a combination thereof.

[0015] Optionally, the causes of the occurrence and development of acne include, but are not limited to: excessive secretion of sebum from sebaceous glands, abnormal keratinization of the hair follicle duct opening, excessive proliferation of Propionibacterium acnes around sebaceous gland tissue and the resulting inflammatory response.

[0016] Optionally, preventing and / or treating acne includes: preventing the formation of acne, improving or alleviating acne symptoms, accelerating the regression or healing of acne, improving acne scars (acne marks), or accelerating the regression of acne scars (acne marks).

[0017] In one embodiment, the present invention provides a method for preventing and / or treating acne with a mulberry extract, comprising administering a product comprising a mulberry extract as an active ingredient to a subject in need thereof, the subject including a human being.

[0018] In one embodiment, the present invention provides a use of a mulberry extract in preparing a product for preventing and / or treating acne.

[0019] In one embodiment of the present invention, the prevention and / or treatment of acne comprises reducing the GAGS score of the patient.

[0020] In one embodiment of the present invention, the prevention and / or treatment of acne includes at least one of inhibiting lipid secretion and accumulation in sebaceous glands, inhibiting abnormal keratinization of hair follicle duct openings, inhibiting excessive proliferation of Propionibacterium acnes around sebaceous gland tissue, and inhibiting inflammatory response of sebaceous gland tissue.

[0021] In one embodiment of the present invention, the prevention and / or treatment of acne comprises any one or more of inhibiting lipid accumulation in sebaceous glands and / or inhibiting inflammatory response of sebaceous gland tissue by regulating sex hormone levels.

[0022] Preferably, the lipid accumulation comprises lipid accumulation in sebaceous gland tissue and / or sebaceous gland cells; preferably, the lipid comprises triglycerides (TG) and free fatty acids (FFA).

[0023] Preferably, the sex hormone level includes the sex hormone level of sebaceous gland tissue; preferably, the sex hormone includes any one of testosterone (T), dihydrotestosterone (DHT), and estradiol (E2).

[0024] Preferably, the inhibiting of the inflammatory response of sebaceous gland tissue comprises regulating the expression of inflammatory response-related proteins in serum and / or sebaceous gland tissue and / or increasing the antioxidant level in serum and / or sebaceous gland tissue.

[0025] Preferably, the inflammatory response-related proteins may include matrix metalloproteinases MMP, preferably MMP-2 and / or MMP-9; preferably, the regulating the expression of inflammatory response-related proteins in serum and / or sebaceous gland tissue includes reducing the expression of MMP-2 and / or MMP-9 in serum and / or sebaceous gland tissue.

[0026] Preferably, said increasing the antioxidant level in serum and / or sebaceous gland tissue comprises increasing the activity of SOD in serum and / or sebaceous gland tissue.

[0027] The inhibiting of acne pathogenic bacteria includes inhibiting excessive proliferation of acne pathogenic bacteria. Preferably, the acne pathogenic bacteria are Propionibacterium acnes.

[0028] Preferably, the prevention and / or treatment of acne comprises any one or more of the following:

[0029] (1) Reduce the patient's GAGS score;

[0030] (2) inhibiting sebum secretion and lipid accumulation of sebaceous glands; preferably, the lipid accumulation includes lipid accumulation in sebaceous gland tissue and / or sebaceous gland cells; preferably, the lipids include triglycerides (TG) and / or free fatty acids (FFA);

[0031] (3) regulating the sex hormone level of sebaceous gland tissue; preferably, any one or more of the following:

[0032] (i) Reduce testosterone (T) and dihydrotestosterone (DHT) levels;

[0033] (ii) reducing the free testosterone index (FAI); wherein FAI=free testosterone concentration (T) / sex hormone binding globulin concentration (SHBG);

[0034] (iii) increase estradiol (E2) content;

[0035] (4) inhibit abnormal keratinization of the hair follicle duct;

[0036] (5) Inhibit the excessive proliferation of Propionibacterium acnes around sebaceous gland tissue;

[0037] (6) Inhibiting the inflammatory response of sebaceous gland tissue, including regulating the expression of inflammatory response-related proteins in serum and / or sebaceous gland tissue and / or increasing the antioxidant level in serum and / or sebaceous gland tissue; preferably, including reducing the expression of MMP-2 and / or MMP-9 in serum and / or sebaceous gland tissue, and / or increasing the activity of SOD in serum and / or sebaceous gland tissue.

[0038] In one embodiment, the novel use of the mulberry extract includes at least one of the following (b1)-(b6):

[0039] (b1) Use of mulberry extract in the preparation of a product for reducing GAGS score in a patient;

[0040] (b2) Use of mulberry extract in the preparation of a product for inhibiting sebum secretion and lipid accumulation in sebaceous glands; preferably, the lipid accumulation includes lipid accumulation in sebaceous gland tissue and / or sebaceous gland cells; preferably, the lipid includes triglycerides (TG) and / or free fatty acids (FFA);

[0041] (b3) Use of mulberry extract in the preparation of a product for regulating sex hormone levels in sebaceous gland tissue; preferably, any one or more of the following:

[0042] (i) Use of mulberry extract in the preparation of a product for reducing the content of testosterone (T) and dihydrotestosterone (DHT) in sebaceous gland tissue;

[0043] (ii) Use of mulberry extract in the preparation of a product for reducing the free testosterone index (FAI) of sebaceous gland tissue;

[0044] (iii) use of mulberry extract in the preparation of a product for increasing the estradiol (E2) content in sebaceous gland tissue;

[0045] (b4) Use of mulberry extract in the preparation of a product for inhibiting abnormal keratinization of hair follicle ducts;

[0046] (b5) Use of mulberry extract in the preparation of a product for inhibiting excessive proliferation of Propionibacterium acnes;

[0047] (b6) Use of mulberry extract in the preparation of a product for inhibiting the inflammatory response of sebaceous gland tissue. Preferably, the inhibiting the inflammatory response of sebaceous gland tissue comprises regulating the expression of inflammatory response-related proteins in serum and / or sebaceous gland tissue and / or increasing the antioxidant level in serum and / or sebaceous gland tissue; preferably, comprises reducing the expression of MMP-2 and / or MMP-9 in serum and / or sebaceous gland tissue, and / or increasing the activity of SOD in serum and / or sebaceous gland tissue.

[0048] In one embodiment of the present invention, the acne is caused by at least one of excessive sebum secretion from sebaceous glands, abnormal keratinization of the hair follicle duct opening, excessive proliferation of Propionibacterium acnes around sebaceous gland tissue, and / or inflammatory response of sebaceous gland tissue. Preferably, the acne is acne caused by excessive sebum secretion from sebaceous glands. Preferably, the acne is acne caused by excessive androgen secretion. Preferably, the acne is acne caused by excessive proliferation of Propionibacterium acnes around sebaceous gland tissue. Preferably, the acne is acne caused by inflammatory response of sebaceous gland tissue.

[0049] Preferably, the acne is located on the face, neck, chest and / or back where sebaceous glands are abundant; preferably the face. Preferably, the face includes the forehead, left and right cheeks and nose; the back is the upper back.

[0050] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0051]

[0052] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0053]

[0054] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0055]

[0056] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0057]

[0058] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0059]

[0060] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0061]

[0062] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0063]

[0064] Further preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0065]

[0066] Further preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0067]

[0068] More preferably, based on the sum of the weight percentage of the components of the mulberry extract being 100%, the weight content of the components in the mulberry extract is:

[0069]

[0070] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0071]

[0072] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0073]

[0074]

[0075] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0076]

[0077] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0078]

[0079] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0080]

[0081] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0082]

[0083]

[0084] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0085]

[0086] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0087]

[0088] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0089]

[0090] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0091]

[0092] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0093]

[0094] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0095]

[0096] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0097]

[0098] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0099]

[0100] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0101]

[0102]

[0103] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0104]

[0105] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0106]

[0107] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0108]

[0109] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0110]

[0111]

[0112] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0113]

[0114] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0115]

[0116] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0117]

[0118] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0119]

[0120] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0121]

[0122] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0123]

[0124] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0125]

[0126] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0127]

[0128] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0129]

[0130] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0131]

[0132] Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0133]

[0134] Preferably, the alkaloids include 1-deoxynojirimycin (DNJ), N-methly-1-deoxynojirimycin, fagomine (FAG), 3-epi-fagomine, 1,4-dideoxy-1,4-imino-D-arabinitol (DAB), calystegin B2, calystegin C1, 2-oxo-(α-D- One or more of 2-O-(α-D-galactopyranosyl)-1-deoxynojirimycin, 6-O-(β-D-glucopyranosyl)-1-deoxynojirimycin, and 1,4-dideoxy-1,4-imino-(2-O-β-D-glucopyranosyl)-D-arabinitol;

[0135] Preferably, the weight percentage of DNJ is not less than 50% (optionally 60-99%) of the total alkaloids.

[0136] Preferably, the heavy metal content of the mulberry extract does not exceed 10 ppm.

[0137] Preferably, the mulberry extract is mulberry extract, mulberry branch extract, mulberry bark extract and / or mulberry leaf extract.

[0138] Preferably, the product for preventing and / or treating acne is selected from at least one of food, medicine, beverage, health product or cosmetics; preferably, the product is a functional additive.

[0139] Preferably, the mulberry extract is applied to humans or mammals.

[0140] Preferably, the product is an oral dosage form or an external dosage form.

[0141] Preferably, the product is a tablet, capsule, oral solution, oral emulsion, pill, granule, syrup or powder.

[0142] Preferably, the product is a lotion, ointment, cream, gel, tincture, liniment, spirit, powder, oil, paste, plaster, film or aerosol, etc., preferably a gel. Preferably, the lotion is a solution type lotion, a suspension type lotion or an emulsion type lotion.

[0143] In the present invention, the mulberry extract can be provided in the form of commercially available mulberry branch total alkaloid tablets (National Medicine Standard No. Z20200002).

[0144] Preferably, the mulberry extract can be prepared according to the method described in CN110393738A.

[0145] Preferably, in the present invention, the preparation of the mulberry extract comprises the following steps:

[0146] 1) preparing a crude extract of a moraceae plant; 2) separating the crude extract through a cationic resin and / or an optional anionic resin to obtain a resin effluent; optional step 3): subjecting the resin effluent of step 2) to alcohol precipitation and collecting the supernatant; 4) concentrating and / or drying the supernatant. Optionally, before subjecting the resin effluent of step 2) to alcohol precipitation, the resin effluent is concentrated and / or dried.

[0147] Preferably, the extraction method further comprises: before the separation in step 2), the crude plant extract is concentrated; preferably, the crude plant extract is concentrated to a mass concentration of solids in the solution of 1-15%, preferably 2-10%. The solids refer to the solid matter remaining after the water in the solution is removed.

[0148] Optionally, the crude extract after the concentration treatment can also be subjected to alcohol precipitation treatment before the resin separation treatment in step 2). During the alcohol precipitation treatment, ethanol is added to the crude extract, stirred and mixed, and the stirring is stopped and allowed to stand for a certain period of time to precipitate the insoluble matter therein. Preferably, in terms of L / kg, the volume mass ratio of the added ethanol to the plant raw material is 0.2-20 times, and 0.4-10 times can be selected. Further preferably, an alcohol precipitation tank is used for alcohol precipitation treatment. Preferably, the stirring speed in the alcohol precipitation treatment is 10-600rpm, optionally 40-500rpm, and further optionally 80-400rpm or 300rpm.

[0149] Preferably, the moraceae plant is Morus multicaulis Perrott., Morus alba L., Morus atropurpurea Roxb., Morus mizuho Hotta, Morus wittiorum Hand Mazz., Morus laevigata Wall, Morus nigra Linn., Morus cathayana Hemsi., Morus serrata Roxb., Morus mongolica Schneid., Morus bombycis Koidz., Morus notabilis Schneid., Morus nigriformis Koidz., Morus yunnanensis Koidz., Morus australis Poir., Morus mongolica (Bur.) Schneid var. diabolica Koidz.), Morus alba Var. Pendula Dippel, Morus alba Var. Pendula Dippel, Morus alba, and mulberry varieties bred from the above mulberry species, and a combination of one or more of the above mulberry species bred intraspecifically or interspecifically;

[0150] Preferably, the moraceae plant is one or more selected from Guangdong mulberry, Shandong mulberry, white mulberry, fine-toothed mulberry, mountain mulberry or hybrid mulberry, and the hybrid mulberry can be selected from Yuesang No. 11, Guisang You No. 62 or Sang Te You No. 2. The leaves, roots, branches, bark, buds, stems, fruits and other parts of the moraceae plant can be used, and mulberry branches, mulberry leaves or white mulberry bark can be used.

[0151] In the present invention, the mulberry extract can be selected from mulberry branch extract, white mulberry bark extract, mulberry leaf extract or a mixed extract thereof.

[0152] Preferably, the moraceae plant is crudely extracted using a solvent such as alcohol water, water, alkaline aqueous solution or acidic aqueous solution. During the extraction, the plant is preferably crushed and added to a solvent for hot extraction. Optionally, the extraction is repeated 1-3 times, and the extraction time is preferably 0.5-3 hours each time, and further preferably 1-3 hours each time.

[0153] In a preferred embodiment, the crushed plants can be added into an extraction tank for extraction.

[0154] Preferably, the extract is filtered to remove insoluble matter to obtain a crude plant extract.

[0155] In one embodiment, the mulberry extract is prepared according to the following steps: crushing the moraceae plant, heating and refluxing extraction with water and / or alcohol solution or acid water, the amount of solvent is 3-20 times (optionally 4-15 times) of the original medicinal material, repeating the extraction 1-3 times (the extraction time can be 0.5-3h each time, and further can be 1-3h each time), combining the extracts, concentrating, applying a cation exchange resin, eluting with 0.2-3N ammonia water, applying the eluate to an anion exchange resin, collecting the non-adsorbed part, adding ethanol, precipitating to remove impurities, concentrating and / or drying to obtain an extract.

[0156] In one embodiment, the mulberry extract is prepared according to the following steps: crushing the moraceae plant, heating and refluxing extraction with water and / or alcohol solution or acid water, the amount of solvent is 3-20 times (preferably 4-15 times) of the original medicinal material, repeating the extraction 1-3 times (the extraction time can be selected as 0.5-3h each time, and further can be selected as 1-3h each time), combining the extracts, concentrating, applying a cation exchange resin, eluting with 0.2-3N ammonia water, applying the eluate to an anion exchange resin, collecting the non-adsorbed part, concentrating and / or drying to obtain an extract.

[0157] In one embodiment, the mulberry extract is prepared according to the following steps: crushing the moraceae plant, heating and refluxing extraction with water and / or alcohol solution or acid water, the amount of solvent is 3-20 times (optionally 4-15 times, further optionally 4-12 times) of the original medicinal material, repeating the extraction 1-3 times (the extraction time can be optionally 0.5-3h each time, further optionally 1-3h each time), combining the extracts, concentrating, applying a cation exchange resin, eluting with 0.2-3N ammonia water, concentrating and / or drying the eluate to obtain an extract.

[0158] Preferably, after the cationic resin is loaded into the column, it is activated in the order of washing with an acidic solution, washing with an alkaline solution, and washing with an acidic solution. Preferably, the alkaline solution is washed until the pH of the eluate is 8.0-9.5, and can be 8.5-9.5; preferably, the alkaline solution is selected from an aqueous ammonia solution, a sodium hydroxide solution, a potassium hydroxide solution, or a sodium carbonate solution; preferably, the concentration of the alkaline solution is 0.5-4 mol / L, and can be 1-2 mol / L. Preferably, the acidic solution is washed until the pH of the eluate is 3.0-7.0, and can be 4.5-6.5. Preferably, the acidic solution is selected from a hydrochloric acid solution, a phosphoric acid solution, and a sodium hydrogen phosphate-citric acid buffer solution. Preferably, the concentration of the acidic solution is 0.5-4 mol / L, and can be 1.5-2 mol / L. Optionally, the cationic resin can also be rinsed with 3-5 column volumes of deionized water after the last acidic solution wash.

[0159] Preferably, the cationic resin is selected from a combination of one or more of a strongly acidic cation exchange resin, a weakly acidic cation exchange resin and a strongly basic quaternary ammonium cation exchange resin. Preferably, the cationic resin is a combination of one or more of a 732 type strongly acidic styrene cation exchange resin, a 002SC type strongly acidic styrene cation exchange resin, a 734 type strongly acidic styrene cation exchange resin, a D001 type macroporous strongly acidic styrene cation exchange resin or a D113 type macroporous weakly acidic cation exchange resin and a D254 type macroporous strongly basic quaternary ammonium cation exchange resin. Preferably, the cationic resin is a combination of one or more of a 732 type strongly acidic styrene cation exchange resin, a 734 type strongly acidic styrene cation exchange resin, a 002SC type strongly acidic styrene cation exchange resin, a D113 type macroporous weakly acidic cation exchange resin and a D001 type macroporous strongly acidic styrene cation exchange resin. Preferably, the cationic resin is a combination of one or more of 002SC type strongly acidic styrene-based cationic resin, D113 type macroporous weakly acidic cationic resin and D001 type macroporous strongly acidic styrene-based cation exchange resin.

[0160] Preferably, the weight ratio of the cationic resin to the plant raw material is 1:1-30 (preferably, 1:1-25, 1:2-20, 1:2-15, 1:2-10, 1:2-7, 1:2-3, 1:4-15).

[0161] After the plant crude extract is loaded onto the cationic resin, the loaded cationic resin is eluted with an eluent. Preferably, the eluent is a salt solution or alkaline solution containing cations, and can be selected from one or more of sodium chloride, ammonium chloride, ammonium sulfate, ammonium nitrate, ammonia water, potassium chloride and sodium hydroxide.

[0162] Preferably, the concentration of the cation in the eluent is 0.04-5 mol / L (optionally 0.5-2.5 mol / L, 0.2-3 mol / L, further optionally 0.5-2.5 mol / L).

[0163] Preferably, the eluent flow rate is 1-15 BV / h (optionally 5-10 BV / h, further optionally 5-6 BV / h).

[0164] Preferably, the weight of the eluent used for cationic resin separation is 0.1-30 times the weight of the plant raw material feed, preferably, the eluent is 0.5-10 times the weight of the plant raw material feed, and more preferably, 1-10 times.

[0165] The collection starting point can be determined according to the pH of the effluent of the cationic resin. For example, when an alkaline solution such as ammonia water is used for elution, the eluate is collected when the pH of the effluent of the cationic column is detected to be greater than 7, or the collection starting point of the effluent is determined according to the color development or precipitation reaction; preferably, when the volume of the collected liquid reaches 0.1-10 times (more preferably, 0.6-10 times, or 0.2-5 times) the weight of the plant raw material, the collection is stopped, and the collected liquid is optionally purified by an anion column. In order to improve the separation effect of the cationic resin, the cationic resin can also be used for multiple separations, for example, 2-5 times.

[0166] When purifying by anion column, preferably, after the anion resin is loaded into the column, it is activated in the order of washing with alkaline solution, washing with acidic solution, and washing with alkaline solution.

[0167] Preferably, the alkaline solution is washed until the pH of the eluate is 8.0-9.5, optionally 8.5-9.5;

[0168] Preferably, the alkaline solution is selected from aqueous ammonia solution, sodium hydroxide solution, potassium hydroxide solution or sodium carbonate solution; preferably, the concentration of the alkaline solution is 0.5-4 mol / L, optionally 1-2 mol / L.

[0169] Preferably, the acidic solution is washed until the pH of the eluate is 3.0-7.0, optionally 4.5-6.5. Preferably, the acidic solution is selected from hydrochloric acid solution, phosphoric acid solution, disodium hydrogen phosphate-citric acid buffer, and preferably, the concentration of the acidic solution is 0.5-4 mol / L, optionally 1-2 mol / L.

[0170] Preferably, the anion resin is one or more combinations selected from strongly basic anion exchange resins, weakly basic anion exchange resins or weakly acidic anion exchange resins. Preferably, the anion resin is a combination of one or more of 711-type strongly basic styrene anion resin, 717-type strongly basic styrene anion exchange resin, D201-type macroporous strongly basic styrene anion exchange resin or D218-type macroporous strongly basic acrylic anion exchange resin, D301-G-type macroporous weakly acidic styrene anion exchange resin and D301-type macroporous weakly basic styrene anion exchange resin. Preferably, the anion resin is a combination of one or more of 711-type strongly basic styrene anion resin, 717-type strongly basic styrene anion exchange resin, D201-type macroporous strongly basic styrene anion exchange resin and D218-type macroporous strongly basic acrylic anion exchange resin. Preferably, the anion resin is a combination of one or more of 711 type strong basic styrene type anion resin, D201 type macroporous strong basic styrene type anion exchange resin and D218 type macroporous strong basic acrylic acid type anion exchange resin.

[0171] Preferably, the weight ratio of the anion resin to the plant raw material is 1:1-80 (preferably, 1:1-64, 1:1-32, 1:1-24, 1:5-16, 1:3).

[0172] When the liquid flows out of the anion resin, collection begins. Preferably, when the volume of the collected liquid reaches 0.05-10 times (preferably 0.1-5 times) the weight of the plant raw material, collection is stopped. Optionally, in order to improve the separation effect of the anion resin, the anion resin can also be used for multiple separations, for example, 2-4 separations.

[0173] Preferably, the weight ratio of ethanol used for alcohol precipitation treatment to plant raw material is 1:4-600 (optionally, 1:20-300, further optionally 1:20-200, 1:20-50, 1:40, 1:80, 1:22, 1:200). During the alcohol precipitation treatment, the stirring speed is 10-600rpm (optionally 40-500rpm, 80-400rpm, 300-400rpm). The time of alcohol precipitation treatment is 12-24h.

[0174] Furthermore, before the alcohol precipitation treatment, the anion resin effluent is centrifuged or filtered through a microfiltration membrane to remove impurities, and then concentrated through a reverse ion osmosis membrane. The specific gravity of the concentrated liquid can be 1.0-1.3, and can be 1.1-1.25. The specific gravity refers to the mass ratio of the concentrated liquid to water under the same volume conditions.

[0175] The concentration treatment method of the present invention may include heat concentration, nanofiltration membrane, reverse ion permeation membrane concentration and a combination thereof. Preferably, centrifugation, ultrafiltration membrane filtration or microfiltration membrane filtration is performed to remove impurities before reverse ion permeation membrane and nanofiltration membrane concentration.

[0176] Preferably, the product further comprises a pharmaceutically, food and / or cosmetically acceptable carrier. The carrier is an inactive ingredient that is non-toxic to the human body and conforms to the route or mode of use. The carrier can be a solid or liquid excipient. Solid excipients, for example, include microcrystalline cellulose, mannitol, lactose, pregelatinized starch, low-substituted hydroxypropyl cellulose, cross-linked polyvinylpyrrolidone, sodium carboxymethyl starch, aspartame, calcium hydrogen phosphate, sodium lactate, poloxamer, sodium lauryl sulfate, sodium carboxymethyl cellulose, gelatin, xanthan gum, polyvinylpyrrolidone, starch, magnesium stearate, sodium carboxymethyl starch and talc; liquid excipients, for example, include water, ethanol, syrup and glycerol.

[0177] In the present invention, the mulberry extract is referred to as SZ-A.

[0178] Beneficial effects:

[0179] The present invention provides an application of a mulberry extract in preparing a product for preventing and / or treating acne, which significantly improves the therapeutic effect on acne, reduces the occurrence of adverse reactions, and provides a better solution for the treatment of acne. BRIEF DESCRIPTION OF THE DRAWINGS

[0180] In order to more clearly illustrate the specific embodiments of the present invention, the drawings used in the specific embodiments will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0181] Figure 1 The effects of oral and topical administration on the morphology of sebaceous gland tissue in golden hamsters in Experimental Example 1 of the present invention;

[0182] Figure 2 The effect of oral and topical administration on the growth rate of sebaceous gland spots in golden hamsters in Experimental Example 1 of the present invention;

[0183] Figure 3 and Figure 4 The effect of oral and topical administration on lipid accumulation in sebaceous gland tissue of golden hamsters in Experimental Example 1 of the present invention;

[0184] Figure 5 The effects of oral and topical administration on the content of triglyceride (TG) and free fatty acid (FFA) in the sebaceous gland plaque tissue of golden hamsters in Experimental Example 1 of the present invention;

[0185] Figure 6 The effect of oral and topical administration on the testosterone (T) content in the sebaceous gland plaque tissue of golden hamsters in Experimental Example 1 of the present invention;

[0186] Figure 7 The effect of oral and topical administration on the content of dihydrotestosterone (DHT) in the sebaceous gland plaque tissue of golden hamsters in Experimental Example 1 of the present invention;

[0187] Figure 8 The effects of oral and topical administration on sex hormone binding globulin (SHBG) and free testosterone index (FAI) in sebaceous gland plaque tissue of golden hamsters in Experimental Example 1 of the present invention;

[0188] Fig. 9 The effect of oral and topical administration on the content of estradiol (E2) in the sebaceous gland plaque tissue of golden hamsters in Experimental Example 1 of the present invention;

[0189] Fig.10The effect of oral and topical administration on the expression of matrix metalloenzyme protein 2 (MMP-2) and matrix metalloenzyme protein 9 (MMP-9) in the serum and sebaceous gland plaque tissue of golden hamsters in Experimental Example 1 of the present invention;

[0190] Fig.11 The effect of oral and topical administration on the activity of superoxide dismutase (SOD) in the serum and sebaceous gland plaque tissue of golden hamsters in Experimental Example 1 of the present invention;

[0191] Fig.12 The effects of mulberry extracts prepared in different examples on the growth rate of sebaceous gland spots in Experimental Example 2 of the present invention;

[0192] Fig.13 and Fig.14 This is the effect of the mulberry extract on lipid accumulation in SZ95 cells in Experimental Example 3 of the present invention.

[0193] Fig.15 It is a result diagram of the antibacterial experiment in Experimental Example 4 of the present invention.

[0194] In the figure, compared with the model group, *P<0.05, **P<0.01, ***P<0.001. DETAILED DESCRIPTION

[0195] 1. Preparation Example of Mulberry Extract

[0196] Preparation Example 1

[0197] Take 1000kg of fresh mulberry branches (fine-toothed mulberry branches, Yuesang 11), crush them, add 4000L of water, extract them by heating reflux for 2h, combine the extracts, filter and remove insoluble matter to obtain a crude extract. The crude extract is heat concentrated to a solid mass percentage of 4%, and kept at 50℃ as the loading liquid for the cationic resin column.

[0198] Use 150kg of D113 macroporous weakly acidic phenyl propylene cationic resin to load the column, wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; wash with 1mol / L sodium hydroxide solution until the pH of the eluate is 8.5; wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; then rinse with 5 times the column volume of deionized water to complete the activation. Load the concentrated extract, then elute with 1000L 2.5mol / L ammonia water at a rate of 6BV / h. Collect the eluate when the pH of the cation column effluent is >7. When the collected liquid reaches 900L, stop collecting and directly purify the collected liquid through the anion column.

[0199] Use 62.5 kg of D218 macroporous strongly alkaline acrylic anion resin to load the column, wash with 1.5 mol / L sodium hydroxide solution until the pH of the eluate is 9.0; wash with 1.5 mol / L hydrochloric acid solution until the pH of the eluate is 3.5; wash with 1.5 mol / L sodium hydroxide solution until the pH of the eluate is 9.0; complete activation. Load the collected cationic resin eluate onto the anion resin, and collect the effluent until the effluent reaches 920 L.

[0200] The collected liquid was centrifuged to remove impurities and then concentrated by reverse ion osmosis membrane. The liquid after concentration had a specific gravity of 1.25, and was transferred to an alcohol precipitation tank. 25L of anhydrous ethanol was added at 500rpm. After the addition of ethanol, stirring was stopped, and the alcohol precipitation was carried out for 24 hours. The supernatant was taken and concentrated under reduced pressure to obtain a mulberry branch extract (i.e., mulberry extract SZ-A).

[0201] The mulberry branch extract has 52% by weight of alkaloids, 22% by weight of polysaccharides, 0.8% by weight of flavonoids, and 20% by weight of amino acids. Among the alkaloids, the content of 1-DNJ is 60%, FAG is 17%, and DAB is 15%.

[0202] Preparation Example 2

[0203] Take 10kg of fresh mulberry branches (Sangteyou No. 2), crush them, add 150L of water, add them in 2 times, extract them by decoction for 3h each time, combine the extracts, and filter to remove insoluble matter. The extract is hot concentrated to 8% solid mass percentage, transferred to an alcohol precipitation tank, and 2367.9g of anhydrous ethanol (3L) is added under a stirring paddle of 300rpm. After the addition of ethanol, stop stirring, precipitate with alcohol for 24h, and take the supernatant as the sample liquid for the cationic resin column. Use 002SC type strong acid styrene cationic resin 5kg to load the column, and activate the cationic resin according to the method of Preparation Example 1. Load the extract treated with concentrated alcohol precipitation, then use 100L 5mol / L potassium chloride for elution, the elution rate is 5BV / h, and detect the effluent with 20% silicotungstic acid. Start collecting when a white precipitate is generated, and stop collecting when the collected liquid reaches 25L, and directly purify the collected liquid through an anion column.

[0204] Use 10 kg of 711 type strong basic styrene anion resin to load the column, and activate the anion resin according to the method of Preparation Example 1. Load the collected cationic resin eluate onto the anion resin, and collect the effluent until the effluent reaches 15 L. Load the collected liquid onto the cationic resin again, and separate twice more using the cationic resin and the anionic resin in turn according to the above method.

[0205] The collected liquid obtained after three column separations was centrifuged to remove impurities and then concentrated by reverse ion permeation membrane. The concentrated liquid had a specific gravity of 1.25 and was transferred to an alcohol precipitation tank. 125 g of anhydrous ethanol was added under a stirring paddle of 1000 rpm. After the addition of ethanol, stirring was stopped, and the alcohol was precipitated for 24 hours. The supernatant was taken and concentrated under reduced pressure to obtain an extract. In addition, fresh mulberry bark and mulberry leaf (Sangteyou No. 2) were extracted, and the extraction method and parameters were the same as those in the above method.

[0206] The obtained mulberry branch extract has a mass percentage of alkaloids of 98%, a mass percentage of polysaccharides of 0.2%, a mass percentage of flavonoids of 0.05%, and a mass percentage of amino acids of 0. Among the alkaloids, the content of 1-DNJ is 99%, FAG is 0.5%, and DAB is 0.4%.

[0207] The obtained Morus alba extract has 95% by weight of alkaloids, 2% by weight of polysaccharides, 0.1% by weight of flavonoids, and 1% by weight of amino acids. Among the alkaloids, the content of 1-DNJ is 96%, FAG is 1.5%, and DAB is 1.4%.

[0208] The obtained mulberry leaf extract has 90% by weight of alkaloids, 4% by weight of polysaccharides, 0.1% by weight of flavonoids, and 3% by weight of amino acids. Among the alkaloids, the content of 1-DNJ is 91%, FAG is 3.1%, and DAB is 2.8%.

[0209] Preparation Example 3

[0210] Take 1000kg of fresh mulberry branches (Guangdong mulberry), crush them, add 11500L of water, heat and reflux for 2h, combine the extracts, filter and remove insoluble matter to obtain a crude extract. The crude extract is first centrifuged to remove impurities, and then concentrated with a counter ion permeation membrane to a solid mass percentage of 1%, which is used as the loading solution for the cationic resin column.

[0211] 300 kg of D001 macroporous strong acid styrene cationic resin was loaded into the column, and the cationic resin was activated according to the method of Preparation Example 1. The concentrated crude extract was loaded, and eluted with 5000 L of 0.04 mol / L ammonium nitrate at a rate of 5 BV / h. The effluent was detected with 20% silicotungstic acid. When a white precipitate was generated, collection began, and when the collected liquid reached 1000 L, collection was stopped.

[0212] The collected liquid obtained after the cationic column separation is concentrated by nanofiltration membrane, and the extract is concentrated under reduced pressure to obtain an extract concentrate.

[0213] The obtained mulberry branch extract has 15% by weight of alkaloids, 20% by weight of polysaccharides, 7% by weight of flavonoids, and 45% by weight of amino acids. Among the alkaloids, the content of 1-DNJ is 55%, FAG is 23%, and DAB is 10%.

[0214] Preparation Example 4

[0215] Take 333 kg of dry mulberry branches (Yue Sang No. 11), crush them, add 4000 L of water, and extract them twice by heating reflux method, each time reflux for 1 hour, combine the extracts, filter, and concentrate the extracts to 1 kg of crude drug / L.

[0216] Use 150kg of D113 macroporous weakly acidic phenyl propylene cationic resin to load the column, wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; wash with 1mol / L sodium hydroxide solution until the pH of the eluate is 8.5; wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; then rinse with 5 times the column volume of deionized water to complete the activation. Load the concentrated extract, then elute with 1000L 2.5mol / L ammonia water at a rate of 6BV / h. Collect the eluate when the pH of the cation column effluent is >7. When the collected liquid reaches 900L, stop collecting and directly purify the collected liquid through the anion column.

[0217] Use 125kg of D218 macroporous strongly alkaline acrylic anion resin to load the column, wash with 1.5mol / L sodium hydroxide solution until the pH of the eluate is 9.0; wash with 1.5mol / L hydrochloric acid solution until the pH of the eluate is 3.5; wash with 1.5mol / L sodium hydroxide solution until the pH of the eluate is 9.0; complete activation. Load the collected cationic resin eluate onto the anion resin, and collect the effluent with a pH greater than 8 until the effluent reaches 870L.

[0218] The collected liquid obtained after the anion column separation is filtered through a microfiltration membrane to remove impurities and then concentrated with a counter ion permeation membrane. The concentrated liquid has a specific gravity of 1.1 and is transferred to an alcohol precipitation tank. 15 kg of anhydrous ethanol is added at a stirring paddle of 400 rpm. After the addition of ethanol, stirring is stopped, and the alcohol precipitation is carried out for 24 hours. The supernatant is taken and concentrated under reduced pressure to obtain a mulberry branch extract. Sample content: the mass percentage of alkaloids is 80%, the mass percentage of polysaccharides is 5%, the mass percentage of flavonoids is 0.1%, and the mass percentage of amino acids is 4%. Among the alkaloids, the content of 1-DNJ is 75%, FAG is 12%, and DAB is 10%.

[0219] Preparation Example 5

[0220] Take 400 kg of dry mulberry branches (Yue Sang No. 11), crush them, add 4000 L of water, and extract them twice by heating reflux method, each time reflux for 1 hour, combine the extracts, filter, and concentrate the extracts to 1 kg of crude drug / L.

[0221] Use 62.5 kg of D218 macroporous strong alkaline acrylic anion resin to load the column, wash with 1.5 mol / L sodium hydroxide solution until the pH of the eluate is 9.0; wash with 1.5 mol / L hydrochloric acid solution until the pH of the eluate is 3.5; wash with 1.5 mol / L sodium hydroxide solution until the pH of the eluate is 9.0; complete activation. Load the collected extract concentrate onto the anion resin and collect 40 L of the effluent.

[0222] The collected solution obtained after the anion column separation was filtered through a microfiltration membrane to remove impurities and then concentrated by a counter ion permeation membrane, and further concentrated and dried under reduced pressure to obtain a mulberry branch extract. Sample content: 3% by mass of alkaloids, 70% by mass of polysaccharides, 10% by mass of flavonoids, and 10% by mass of amino acids. Among the alkaloids, the content of 1-DNJ was 68%, FAG was 17%, and DAB was 8%.

[0223] Preparation Example 6

[0224] Take 1500kg of fresh mulberry branches (fine-toothed mulberry branches, Yuesang 11), crush them, add 6000L of water, extract them by heating reflux for 2h, combine the extracts, filter and remove insoluble matter to obtain a crude extract. The crude extract is heat concentrated to a solid mass percentage of 4%, and kept at 50℃ as the loading liquid for the cationic resin column.

[0225] Use 100kg of D113 macroporous weakly acidic phenyl propylene cationic resin to load the column, wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; wash with 1mol / L sodium hydroxide solution until the pH of the eluate is 8.5; wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; then rinse with 5 times the column volume of deionized water to complete the activation. Load the concentrated extract, then elute with 1000L 2.5mol / L ammonia water at a rate of 6BV / h. Collect the eluate when the pH of the cation column effluent is >7. Stop collecting when the collected liquid reaches 900L, and directly purify the collected liquid through the anion column.

[0226] Use 62.5kg of D218 macroporous strong alkaline acrylic anion resin to load the column, use 1.5mol / L sodium hydroxide solution to wash until the pH of the eluate is 9.0; use 1.5mol / L hydrochloric acid solution to wash until the pH of the eluate is 3.5; use 1.5mol / L sodium hydroxide solution to wash until the pH of the eluate is 9.0; complete activation. Load the collected cationic resin eluate onto the anion resin, and collect the effluent until the effluent reaches 870L. Concentrate the effluent under reduced pressure to obtain a mulberry branch extract, in which the mass percentage of alkaloids is 30%, the mass percentage of polysaccharides is 35%, the mass percentage of flavonoids is 2%, and the mass percentage of amino acids is 25%. Among the alkaloids, the content of 1-DNJ is 62%, FAG is 20%, and DAB is 13%.

[0227] Preparation Example 7

[0228] Take 1000kg of fresh mulberry branches (fine-toothed mulberry branches, Yuesang 11), crush them, add 4000L of water, extract them by heating reflux for 2h, combine the extracts, filter and remove insoluble matter to obtain a crude extract. The crude extract is heat concentrated to a solid mass percentage of 4%, and kept at 50℃ as the loading liquid for the cationic resin column.

[0229] Use 100kg of D113 macroporous weakly acidic phenyl propylene cationic resin to load the column, wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; wash with 1mol / L sodium hydroxide solution until the pH of the eluate is 8.5; wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; then rinse with 5 times the column volume of deionized water to complete the activation. Load the concentrated extract, then elute with 1000L 2.5mol / L ammonia water at a rate of 6BV / h. Collect the eluate when the pH of the cation column effluent is >7. Stop collecting when the collected liquid reaches 900L, and directly purify the collected liquid through the anion column.

[0230] Use 62.5kg of D218 macroporous strong alkaline acrylic anion resin to load the column, use 1.5mol / L sodium hydroxide solution to wash until the pH of the eluate is 9.0; use 1.5mol / L hydrochloric acid solution to wash until the pH of the eluate is 3.5; wash with 1.5mol / L sodium hydroxide solution until the pH of the eluate is 9.0; complete activation. Load the collected cationic resin eluate onto the anion resin, and collect the effluent until the effluent reaches 870L. Concentrate the effluent under reduced pressure to obtain a mulberry branch extract, in which the mass percentage of alkaloids is 40%, the mass percentage of polysaccharides is 25%, the mass percentage of flavonoids is 0.5%, and the mass percentage of amino acids is 25%. Among the alkaloids, the content of 1-DNJ is 57%, FAG is 24%, and DAB is 16%.

[0231] Preparation Example 8

[0232] Take 333 kg of dry mulberry branches (Yue Sang No. 11), crush them, add 4000 L of water, and extract them twice by heating reflux method, each time reflux for 1 hour, combine the extracts, filter, and concentrate the extracts to 1 kg of crude drug / L.

[0233] Use 150kg of D113 macroporous weakly acidic phenyl propylene cationic resin to load the column, wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; wash with 1mol / L sodium hydroxide solution until the pH of the eluate is 8.5; wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; then rinse with 5 times the column volume of deionized water to complete the activation. Load the concentrated extract, then elute with 1000L 2.5mol / L ammonia water at a rate of 6BV / h. Collect the eluate when the pH of the cation column effluent is >7. When the collected liquid reaches 900L, stop collecting and directly purify the collected liquid through the anion column.

[0234] Use 62.5kg of D218 macroporous strongly alkaline acrylic anion resin to load the column, wash with 1.5mol / L sodium hydroxide solution until the pH of the eluate is 9.0; wash with 1.5mol / L hydrochloric acid solution until the pH of the eluate is 3.5; wash with 1.5mol / L sodium hydroxide solution until the pH of the eluate is 9.0; complete activation. Load the collected cationic resin eluate onto the anion resin, and collect the effluent with a pH greater than 8 until the effluent reaches 870L.

[0235] The collected liquid obtained after the anion column separation is filtered through a microfiltration membrane to remove impurities and then concentrated with a counter ion permeation membrane. The concentrated liquid has a specific gravity of 1.1 and is transferred to an alcohol precipitation tank. 15 kg of anhydrous ethanol is added at a stirring paddle of 400 rpm. After the addition of ethanol, stirring is stopped, and the alcohol precipitation is carried out for 24 hours. The supernatant is taken and concentrated under reduced pressure to obtain a mulberry branch extract. Sample content: the mass percentage of alkaloids is 63%, the mass percentage of polysaccharides is 23%, the mass percentage of flavonoids is 1%, and the mass percentage of amino acids is 5%. Among the alkaloids, the content of 1-DNJ is 61.9%, FAG is 16.6%, and DAB is 11.1%.

[0236] Preparation Example 9

[0237] Take 1000kg of fresh mulberry branches (Yue Sang No. 11), crush them, add 4000L of water, extract them by heating reflux method for 2h, combine the extracts, filter and remove insoluble matter to obtain crude extract. The crude extract is heat concentrated to a solid content of 4%, and kept at 50℃ as the loading liquid for the cationic resin column.

[0238] Use 120kg of D113 macroporous weakly acidic phenyl propylene cationic resin to load the column, wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; wash with 1mol / L sodium hydroxide solution until the pH of the eluate is 8.5; wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; and then rinse with 5 times the column volume of deionized water to complete the activation. Load the concentrated extract, then elute with 1000L 2.5mol / L ammonia water at a rate of 6BV / h. Collect the eluate when the pH of the cation column effluent is >7. When the collected liquid reaches 900L, stop collecting and directly purify the collected liquid through the anion column.

[0239] Use 45kg of D218 macroporous strongly alkaline acrylic anion resin to load the column, wash with 1.5mol / L sodium hydroxide solution until the pH of the eluate is 9.0; wash with 1.5mol / L hydrochloric acid solution until the pH of the eluate is 3.5; wash with 1.5mol / L sodium hydroxide solution until the pH of the eluate is 9.0; complete activation. Load the collected cationic resin eluate onto the anion resin and collect the effluent until the effluent reaches 870L.

[0240] The collected liquid obtained after separation by anion column is filtered through a microfiltration membrane to remove impurities and then concentrated with a counter ion permeation membrane. The concentrated liquid has a specific gravity of 1.1 and is transferred to an alcohol precipitation tank. 15 kg of anhydrous ethanol is added at 300 rpm. After the addition of ethanol, stirring is stopped, and the alcohol precipitation is carried out for 24 hours. The supernatant is taken and concentrated under reduced pressure to obtain an extract. Sample content: the mass percentage of alkaloids is 70%, the mass percentage of polysaccharides is 20%, the mass percentage of flavonoids is 0.6%, and the mass percentage of amino acids is 5%. Among the alkaloids, the content of 1-DNJ is 70%, FAG is 13%, and DAB is 10%.

[0241] Preparation Example 10

[0242] Take 80kg of fresh mulberry branches (Guisangyou No. 62), crush them, add 1000L of water, extract them by heating reflux method for 2h, combine the extracts, filter and remove insoluble matter to obtain crude extract. The crude extract is heat concentrated to a solid content of 4%, and kept at 50℃ as the loading liquid for the cationic resin column.

[0243] Use 50kg of D113 macroporous weakly acidic phenyl propylene cationic resin to load the column, wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; wash with 1mol / L sodium hydroxide solution until the pH of the eluate is 8.5; wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; and then rinse with 5 times the column volume of deionized water to complete the activation. Load the concentrated extract, then elute with 600L2.5mol / L ammonia water at a rate of 6BV / h. Collect the eluate when the pH of the cation column effluent is >7. When the collected liquid reaches 800L, stop collecting and directly purify the collected liquid through the anion column.

[0244] Use 65kg of D218 macroporous strongly alkaline acrylic anion resin to load the column, wash with 1.5mol / L sodium hydroxide solution until the pH of the eluate is 9.0; wash with 1.5mol / L hydrochloric acid solution until the pH of the eluate is 3.5; wash with 1.5mol / L sodium hydroxide solution until the pH of the eluate is 9.0; complete activation. Load the collected cationic resin eluate onto the anion resin and collect the effluent until the effluent reaches 750L. Reload the collected liquid onto the cationic resin, and separate it again using the cationic resin and anionic resin in turn according to the above method.

[0245] The collected liquid obtained after two separations is filtered through a microfiltration membrane to remove impurities and then concentrated with a counter ion permeation membrane. The concentrated liquid has a specific gravity of 1.1 and is transferred to an alcohol precipitation tank. 400g of anhydrous ethanol is added at 350rpm. After the addition of ethanol, stirring is stopped, and the alcohol precipitation is carried out for 24 hours. The supernatant is taken and concentrated under reduced pressure to obtain an extract. Sample content: the mass percentage of alkaloids is 90%, the mass percentage of polysaccharides is 3%, the mass percentage of flavonoids is 0.5%, and the mass percentage of amino acids is 2%. Among the alkaloids, the content of 1-DNJ is 94%, FAG is 2%, and DAB is 1%.

[0246] 2. Efficacy test of mulberry extract

[0247] Experimental Example 1: Acne Treatment Effect of Mulberry Extract in Animal Models

[0248] Golden hamsters are an ideal animal model for verifying the therapeutic effects of acne. This experimental example investigated the therapeutic effect of mulberry extract in this model.

[0249] 1 Experimental plan

[0250] 1.1 Experimental animals

[0251] 6-week-old LVG Hamster male golden hamsters (weight 100±20g, SPF grade, purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., license number SCXK (Beijing) 2021-0011) were kept at 23±2℃, relative humidity 55±5%, diurnal change of 12h, and good ventilation conditions. After 7 days of adaptive feeding with complete nutritional pellet feed, those with no adverse reactions, normal diet, drinking water, and activities were included in the experiment, placed in separate cages, and allowed to drink water freely.

[0252] 1.2 Experimental drugs

[0253] This experiment used the mulberry branch extract (SZ-A) in Preparation Example 9

[0254] Oral medications:

[0255] Morus truncatula low-dose group (SZ-A 100 mg / kg): prepared daily, weighed 147 mg SZ-A, dissolved in 4.853 ml normal saline, and the drug was administered by intragastric administration after it was completely dissolved;

[0256] Morus alba high-dose group (SZ-A 200 mg / kg): daily preparation, weigh 294 mg SZ-A, dissolve in 4.706 ml normal saline, and after complete dissolution, the drug was administered by intragastric administration;

[0257] Topical medications:

[0258] SZ-A gel (SZ-A@gel), preparation method: 1) weigh 2.0g of carbomer and dissolve it in 98mL of normal saline, pre-swell for 24h to obtain 2% carbomer gel; 2) weigh an appropriate amount of SZ-A and dissolve it in 29mL of normal saline, add glacial acetic acid to adjust the pH to 6.6-6.8; mix the carbomer gel and SZ-A solution in a volume ratio of 1:1, and finally control the pH to 5.5-6.0 to obtain 60mg / ml (calculated as alkaloid) SZ-A gel.

[0259] 1.3 Experimental groups and drug administration

[0260] 49 golden hamsters were randomly divided into 7 groups, 7 in each group, and housed individually in single cages. The grouping and drug administration are shown in Table 1.

[0261] Table 1

[0262]

[0263] 1.4 Detection indicators and detection methods

[0264] 28 days after administration, serum and sebaceous gland plaque tissues of golden hamsters were collected for detection of relevant indicators.

[0265] (1) Observation of sebaceous gland tissue morphology and sebaceous gland plaque growth rate: The morphological changes of the sebaceous glands of golden hamsters were monitored during the continuous drug administration period. Photos were taken with a camera on days 0, 7, 14, 21, and 28 of drug administration, and the area of ​​the sebaceous gland plaques was calculated. ImageJ was used for data processing.

[0266] (2) Lipid accumulation in sebaceous gland tissue: After 28 consecutive days of drug administration, sebaceous gland tissues were collected from golden hamsters and stained with Oil Red O. The lipid accumulation in sebaceous gland tissues was observed under a microscope and statistical data were collected.

[0267] (3) Triglyceride (TG) and free fatty acid (FFA) content: The TG content in the sebaceous gland plaque tissue of golden hamsters was determined by the GPO-PAP method according to the instructions of the kit; the FFA content in the sebaceous gland plaque tissue of golden hamsters was determined by the enzymatic method according to the instructions of the kit.

[0268] (4) Sex hormone (testosterone (T), sex hormone binding globulin (SHBG), dihydrotestosterone (DHT), estradiol (E2)) levels: The levels of T, SHBG, DHT, and E2 in the sebaceous gland tissue of golden hamsters were determined by enzyme-linked immunosorbent assay according to the instructions of the kit, and the FAI was calculated (FAI = T / SHBG).

[0269] (5) Expression levels of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9): The levels of MMP-2 and MMP-9 in the serum and sebaceous gland tissues of golden hamsters were determined by enzyme-linked immunosorbent assay according to the instructions of the kit.

[0270] (6) Superoxide dismutase (SOD) activity: The activity of SOD in the serum and sebaceous gland tissue of golden hamsters was determined according to the instructions of the kit.

[0271] 2. Test results

[0272] (1) Observation of sebaceous gland tissue morphology and sebaceous gland spot growth rate

[0273] The area of ​​sebaceous gland spots in golden hamsters is the most intuitive reflection of drug treatment. The morphology of sebaceous gland spots was observed and the growth rate was calculated on days 0, 7, 14, 21 and 28 of drug administration:

[0274] 1) The results of sebaceous gland tissue morphology observation are as follows Figure 1 shown.

[0275] Oral administration: Compared with the Model group, after 28 days of administration, the area of ​​sebaceous gland spots in the positive group, SZ-A100mg and SZ-A200mg groups were reduced, and the color became lighter;

[0276] For external use: Compared with the Model group, after 28 days of administration, the sebaceous gland spots in the positive group and the SZ-A gel group became lighter and the raised area of ​​the spots was reduced.

[0277] 2) Sebaceous gland spot growth rate test results Figure 2 shown.

[0278] Oral administration: Compared with the Model group, SZ-A100mg and SZ-A200mg had a tendency to reduce the growth rate of sebaceous gland spots after 7 and 14 days of administration, and could significantly reduce the growth rate of sebaceous gland spots after 21 and 28 days of administration;

[0279] Topical application: Compared with the Model group, topical application of SZ-A@gel could significantly reduce the growth rate of sebaceous gland spots at all time points.

[0280] (2) Lipid accumulation in sebaceous gland tissue

[0281] Excessive sebum secretion can lead to excessive keratinization of the stratum corneum, which in turn induces inflammation and forms acne. After 28 days of administration, lipid accumulation was observed using Oil Red O staining. The results are as follows Figure 3 and Figure 4 shown.

[0282] After 28 days of oral administration: Compared with the Model group, both SZ-A100mg and 200mg showed a trend of reducing lipid accumulation in sebaceous gland tissue, and the therapeutic effect of SZ-A100mg was better than that of the positive drug;

[0283] After 28 days of topical administration: Compared with the Model group, SZ-A@gel could reduce the accumulation of lipids in the sebaceous gland tissue.

[0284] (3) Triglyceride (TG) and free fatty acid (FFA) content

[0285] The results of the detection of triglyceride (TG) and free fatty acid (FFA) content in sebaceous gland tissue are as follows Figure 5 shown.

[0286] After 28 days of oral administration: In the sebaceous glands, TG and FFA showed a decreasing trend compared with the Model group, and the therapeutic effect of FFA in the SZ-A200mg group was better than that of the positive drug, which was statistically significant.

[0287] After 28 days of topical administration: In the sebaceous glands, compared with the Model group, both SZ-A@gel and the positive drugs showed a trend of reducing TG and FFA, and the therapeutic effect of SZ-A@gel was better than that of the positive drugs, with a statistically significant difference.

[0288] (4) Sex hormone levels: testosterone (T), dihydrotestosterone (DHT), sex hormone binding globulin (SHBG), free testosterone index (FAI, FAI = T / SHBG), estradiol (E2)

[0289] Sex hormone imbalance is one of the causes of acne. The development of sebaceous glands and sebum secretion are regulated by hormones. Testosterone (T) is the main androgen. Under the action of 5α-reductase, it produces more active dihydrotestosterone (DHT), which can increase sebum secretion. However, when androgens bind to androgen binding globulin (SHBG), they lose their activity. Estradiol (E2) is a highly biologically active estrogen that can reduce sebum secretion.

[0290] 1) Testosterone (T) level test results Figure 6 shown.

[0291] Oral administration for 28 days: In sebaceous glands, compared with the Model group, both SZ-A100mg and SZ-A200mg showed a dose-dependent trend to reduce T. The positive control and SZ-A200mg groups had similar therapeutic effects.

[0292] Topical administration for 28 days: In sebaceous glands, SZ-A@gel showed a tendency to reduce T compared with the Model group.

[0293] 2) Dihydrotestosterone (DHT) level test results Figure 7 shown.

[0294] Oral administration for 28 days: In the sebaceous glands, compared with the Model group, both the positive drug and SZ-A200mg showed a trend of reducing DHT, and the therapeutic effect of SZ-A200mg was better than that of the positive drug, with a statistically significant difference.

[0295] Topical administration for 28 days: In sebaceous glands, SZ-A@gel showed a tendency to reduce DHT compared with the Model group.

[0296] 3) Sex hormone binding globulin (SHBG) and free testosterone index (FAI) test results Figure 8 shown.

[0297] Oral administration for 28 days: Compared with Model, SZ-A can increase SHBG in sebaceous glands and decrease FA1 in sebaceous glands.

[0298] Topical administration for 28 days: Compared with the Model, SZ-A@gel can increase SHBG in sebaceous glands and tend to reduce FAl in sebaceous glands.

[0299] 4) Estradiol (E2) level test results Fig. 9 shown.

[0300] Oral administration for 28 days: In sebaceous glands, compared with the Model group, SZ-A100mg showed a trend of significantly increasing E2 (better than the positive drug), with statistical difference;

[0301] Topical administration for 28 days: In sebaceous glands, SZ-A@gel showed a tendency to increase E2 compared with the Model group, with no significant changes.

[0302] 5) MMP-2 / MMP-9 expression levels

[0303] Matrix metalloproteinases are associated with acne inflammatory response. The expression levels of MMP-2 / MMP-9 are shown in Fig.10 shown.

[0304] Oral administration for 28 days: In serum, compared with the Model group, SZ-A reduced the expression of MMP-9, and SZ-A200mg significantly reduced it, with statistical significance; in sebaceous glands, compared with the Model group, SZ-A tended to reduce MMP-2; SZ-A200mg tended to significantly reduce MMP-9 (the therapeutic effect was equivalent to that of the positive drug), which was dose-dependent.

[0305] Topical administration for 28 days: In the sebaceous glands, compared with the Model group, MMP-2 was significantly reduced (slightly lower than the positive drug), and MMP-9 also showed a significant downward trend (equivalent to the positive drug), with statistical differences.

[0306] 6) SOD activity

[0307] Oxidative stress is one of the characteristics of acne patients. The total antioxidant level of patients is lower than that of normal people. After 28 days of administration, the results of SOD activity test in serum and sebaceous glands are as follows Fig.11 shown.

[0308] Oral administration: Compared with the Model group, SZ-A showed a tendency to increase SOD in serum and sebaceous glands, but there was no significant change.

[0309] External use: Compared with the Model group, SZ-A@gel has a tendency to increase SOD activity in the sebaceous glands, and the therapeutic effect is better than that of positive drugs.

[0310] The overall results showed that SZ-A had a good effect on acne-related indicators regardless of oral or topical administration.

[0311] Experimental Example 2: Acne treatment effects of mulberry extracts prepared in different examples in animal models

[0312] 1 Experimental plan

[0313] 1.1 Experimental animals

[0314] Same as Experimental Example 1.

[0315] 1.2 Experimental drugs

[0316] The preparation method of 2% carbomer gel is as follows: 2.0 g of carbomer is weighed and dissolved in 98 mL of normal saline, and pre-swelled for 24 hours to obtain 2% carbomer gel;

[0317] Blank gel group: Take the above 2% carbomer gel, dilute it by half with physiological saline, stir well to form a gel.

[0318] The mulberry extract gel SZ-A@gel was prepared using the mulberry branch extracts with different alkaloid contents in the preparation example as follows:

[0319] 1) 15% SZ-A@gel: Weigh 4.8 g SZ-A (mulberry extract in Preparation Example 3), dissolve it in 40 ml physiological saline, adjust the pH to 6.6-6.8 with glacial acetic acid, add 40 g 2% carbomer gel, mix well, and obtain.

[0320] 2) 30% SZ-A@gel: Weigh 4.8 g of SZ-A (the mulberry extract in Preparation Example 6), dissolve it in 40 ml of physiological saline, adjust the pH to 6.6-6.8 with glacial acetic acid, add 40 g of 2% carbomer gel, mix well, and obtain.

[0321] 3) 52% SZ-A@gel: Weigh 4.8 g of SZ-A (the mulberry extract in Preparation Example 1), dissolve it in 40 ml of physiological saline, adjust the pH to 6.6-6.8 with glacial acetic acid, add 40 g of 2% carbomer gel, mix well, and obtain.

[0322] 4) 90% SZ-A@gel: Weigh 4.8 g of SZ-A (the mulberry extract in Preparation Example 10), dissolve it in 40 ml of physiological saline, adjust the pH to 6.6-6.8 with glacial acetic acid, add 40 g of 2% carbomer gel, mix well, and obtain.

[0323] 1.3 Experimental groups and drug administration

[0324] 35 golden hamsters were randomly divided into 7 groups, 5 in each group, and housed individually in individual cages. The detailed grouping and drug administration are shown in Table 2 below.

[0325] Table 2

[0326]

[0327] 1.4 Detection indicators and detection methods

[0328] Observation of sebaceous gland plaque morphology and growth rate: The changes of sebaceous gland plaques of golden hamsters were monitored during the continuous administration period. Photos were taken with a camera on days 0, 7, 14, 21, and 28, and the area of ​​sebaceous gland plaques was calculated. ImageJ was used for data processing.

[0329] 2. Test results

[0330] Observation on the morphology and growth rate of sebaceous gland spots

[0331] The area of ​​sebaceous glands in golden hamsters is the most intuitive reflection of drug treatment. The morphological observation and growth rate results after 28 days of drug treatment are as follows: Fig.12 shown.

[0332] Topical administration: Compared with the blank group, after 28 days of administration, it can be seen that the higher the alkaloid content of mulberry extract, the more obvious the decrease in the growth rate of sebaceous gland spots, and 15%, 30%, 52%, and 90% mulberry extracts are statistically significant. Positive drug adapalene gel can reduce the growth rate of acne, and has statistical significance.

[0333] The results showed that the SZ-A gel groups prepared with the mulberry extracts in Preparation Examples 1, 3, 6 and 10 could inhibit the growth rate of sebaceous gland spots, and the SZ-A gel was more effective than the blank gel.

[0334] Experimental Example 3: Evaluation of the effect of mulberry extract in inhibiting lipid production in the SZ95 cell model

[0335] 1 Experimental plan

[0336] 1.1 Experimental Materials

[0337] SZ95 human sebaceous gland cells.

[0338] 1.2 Experimental groups and drug administration

[0339] SZ95 human sebaceous gland cells, after cell recovery, were cultured in a dedicated DMEM complete medium (BFB, 210B530051, 10% FBS, 90% high glucose basal medium and 1% triple antibody) in an incubator at 37°C, 5% CO2. Cells in the logarithmic growth phase were inoculated into a 24-well plate. After 24 hours of culture, the cells grew completely attached to the wall. Testosterone was added to induce the model, and the mulberry extract in Preparation Example 9 or the gel prepared with the extract was given for 24 hours (the gel preparation method used was the same as that in Experimental Example 1).

[0340] The specific grouping and dosing are shown in Table 3 below.

[0341] Table 3

[0342]

[0343] 1.3 Detection indicators and detection methods

[0344] Lipid accumulation in SZ95 cells: Nile red staining was performed according to the kit instructions to measure the fluorescence intensity in the cells.

[0345] 2. Test results

[0346] Lipid accumulation in SZ95 cells: Sebaceous glands are skin appendages that synthesize sebum, and sebaceous gland cells are used to secrete sebum, which is one of the key factors in the onset of acne. The results of the effect of SZ-A on lipid accumulation in SZ95 cells are as follows: Fig.13 and Fig.14 shown.

[0347] The results showed that compared with the Model, both SZ-A and SZ-A@gel could significantly reduce the accumulation of lipids.

[0348] Experimental Example 4: Antibacterial Effect of Mulberry Extract

[0349] Human skin is a living environment for a variety of microorganisms, and the skin microbiome plays an important role in controlling pathogen colonization and regulating the skin immune system. After the sebaceous glands are stimulated by androgens, they proliferate, and the amount of sebum synthesized and secreted increases and becomes thicker. At the same time, the excessive keratinization of the hair follicle duct narrows the channel for excreting sebum, and the increase in sebum but poor excretion promotes the reproduction of Propionibacterium acnes (P. acnes) in the anaerobic environment, forming an acne pathogenesis with sebum hyperplasia, obstructed sebum excretion, and bacterial infection as the axis. This experimental example investigated the inhibition of mulberry extract on Propionibacterium acnes.

[0350] 1. Antibacterial experiment types, materials and groups

[0351] Detailed antibacterial experimental types, materials and groups are shown in Table 4.

[0352] Table 4

[0353]

[0354] 2. Detection indicators

[0355] Inhibition zone test

[0356] 1) Take out the bacteria frozen in a -80℃ refrigerator, place it on ice, wait for it to thaw, then pipette 10μL into a shaking tube containing 5-6mL MH broth, and shake overnight at 37℃, 200rpm.

[0357] 2) Add 0.1 mL of overnight shaken bacterial solution to a sterile plate, spread evenly, take out a sterile disc filter paper (8 mm in diameter) soaked in the solution for more than 1 hour with tweezers, place it on the plate, cover it, and invert it in a 37°C incubator for 18-24 hours. The solution is the mulberry extract solution in Preparation Example 9. Observe the presence and size of the antibacterial zone around the filter paper disc, and take photos and record.

[0358] 3. Experimental results

[0359] The results of the inhibition zone test are as follows Fig.15 shown.

[0360] The results showed that SZ-A had a significant antagonistic effect on Propionibacterium acnes.

[0361] Experimental Example 5: Clinical Evaluation

[0362] 1. Experimental Protocol

[0363] 1.1 Experimental Grouping

[0364] A randomized, double-blind, double-dummy, placebo-controlled, multicenter study design was adopted. Women of childbearing age aged 18-45 years with acne were included, with a course of 2-8 years. 69 subjects with acne were randomly divided into the Morus truncatus total alkaloids tablets group (SZ-A, N=46) and the placebo group (CON, N=23). All of them signed the informed consent.

[0365] The main exclusion criteria are: 1) skin diseases or other wounds, such as herpes and warts; 2) receiving α-glucosidase inhibitors, or other estrogen, progesterone, lipid-lowering, and hypoglycemic drugs within 12 weeks before the first screening assessment; 3) women who are pregnant, lactating, or unable to take effective contraceptive measures; 4) those with congenital adrenal hyperplasia / hypercortisolism / androgen-secreting tumors / Cushing's syndrome / thyroid dysfunction / hypogonadotropin / hyperprolactinemia / premature ovarian insufficiency / functional hypothalamic-pituitary amenorrhea; 5) those with mental illness and unable to communicate normally.

[0366] 1.2 Administration

[0367] The experimental group took the tablets orally, with the initial dose of 1 tablet each time, 3 times a day; after 4 weeks, the dose was increased to 2 tablets each time, 3 times a day; the control group took the placebo orally, 1 tablet each time, 3 times a day.

[0368] Preparation method of tablets: Take the mulberry branch extract of Preparation Example 8, add appropriate amount of auxiliary materials, mix evenly, add water to make a soft material, granulate and dry, add magnesium stearate, mix evenly, and tablet to obtain a mulberry branch extract preparation, each tablet containing 50 mg of total alkaloids.

[0369] 1.3 Experimental data detection and processing

[0370] 1. At the 12th and 24th weeks of administration, the GAGS scoring standard was used to score 6 areas, including the forehead, left and right cheeks, nose, chest and upper back, with a score of 0-4: acne (1 point), papules (2 points), pustules (3 points) or nodules (4 points). The efficacy index = (GAGS score before treatment - GAGS score after treatment) / GAGS score before treatment.

[0371] 2. Statistical Analysis

[0372] SPSS 20.0 software was used for data processing, and T-test was used to analyze the differences among the groups.

[0373] 2. Experimental results

[0374] Table 5

[0375]

[0376] Remark: * Compared with before treatment, P<0.05; # Comparison between groups p<0.05

[0377] After 12 weeks and 24 weeks of administration, the data of the two groups of acne patients are shown in Table 5 above. The GAGS scores of the subjects in the treatment group SZ-A were significantly improved from 12 weeks to 24 weeks, and there was a significant statistical difference in the efficacy index between the treatment group SZ-A and the control group Con. The efficacy index of 8 of the 46 subjects exceeded 90%, which was considered cured, and the efficacy of the remaining 38 subjects was higher than 60%, which was considered significantly improved.

[0378] In summary, whether in animal experiments, cell experiments, antibacterial experiments or clinical patient experiments, it can be seen that mulberry extract has a good effect on the treatment of acne; at the same time, mulberry extract extracted from natural plants has low side effects and strong patient compliance, providing a better solution for the treatment of acne patients.

Claims

1. Use of mulberry extract in preparing products for preventing and / or treating acne, characterized in that: Taking the weight percentage of the sum of the components of the mulberry extract as 100%, the mulberry extract contains alkaloids with a weight content of more than 3% (optionally containing alkaloids with a weight content of 3-99%, further optionally containing alkaloids with a weight content of 15-99%, further preferably, containing alkaloids with a weight content of 30-99%, further optionally containing alkaloids with a weight content of 40-99%, further optionally containing alkaloids with a weight content of 50-99%, further optionally containing alkaloids with a weight content of 60-99%), and / or comprising a polysaccharide content of not more than 70% by weight (optionally comprising a polysaccharide content of 0.2-70% by weight, further optionally comprising a polysaccharide content of 0.2-50% by weight, further optionally comprising a polysaccharide content of 0.2-35% by weight, further optionally comprising a polysaccharide content of 0.2-25% by weight, further optionally comprising a polysaccharide content of 0.2-23% by weight, further optionally comprising a polysaccharide content of 20-25% by weight), and / or contains flavonoids having a weight content of not more than 10% (optionally containing flavonoids having a weight content of 0.05-5%, further optionally containing flavonoids having a weight content of 0-2%, further optionally containing flavonoids having a weight content of 0.05-2%, further optionally containing flavonoids having a weight content of 0.5-1.5%, further optionally containing flavonoids having a weight content of 0-1%, further optionally containing flavonoids having a weight content of 0.05-1%), and / or contains no more than 50% amino acids by weight (optionally contains 0-30% amino acids by weight, further optionally contains 0-25% amino acids by weight, further optionally contains 0-20% amino acids by weight, further optionally contains 0-5% amino acids by weight, further optionally contains 3-25% amino acids by weight or further optionally contains 5-20% amino acids by weight), and / or other components (the weight content is optionally 0-25%, further optionally 0-20%, further optionally 0-15%, further optionally 0-11%, further optionally 2-20%, further optionally 4-8%).

2. The use according to claim 1, characterized in that: The prevention and / or treatment of acne may include any one or more of the following (1)-(6): (1) Reduce the patient's GAGS score; (2) inhibiting sebum secretion and lipid accumulation of sebaceous glands; preferably, the lipid accumulation includes lipid accumulation in sebaceous gland tissue and / or sebaceous gland cells; preferably, the lipids include triglycerides (TG) and / or free fatty acids (FFA); (3) regulating the sex hormone level of sebaceous gland tissue; preferably, any one or more of the following (i)-(iii): (i) reducing the content of testosterone (T) and dihydrotestosterone (DHT); (ii) reducing the free testosterone index (FAI); wherein FAI = free testosterone concentration (T) / sex hormone binding globulin concentration (SHBG); (iii) increasing the content of estradiol (E2); (4) inhibit abnormal keratinization of the hair follicle duct; (5) Inhibit the excessive proliferation of Propionibacterium acnes around sebaceous gland tissue; (6) Inhibiting the inflammatory response of sebaceous gland tissue, including regulating the expression of inflammatory response-related proteins in serum and / or sebaceous gland tissue and / or increasing the antioxidant level in serum and / or sebaceous gland tissue; preferably, including reducing the expression of MMP-2 and / or MMP-9 in serum and / or sebaceous gland tissue, and / or increasing the activity of SOD in serum and / or sebaceous gland tissue.

3. The use according to claim 1, characterized in that: The application of the mulberry extract is at least one of the following (b1)-(b6): (b1) Use of mulberry extract in the preparation of a product for reducing GAGS score in a patient; (b2) Use of mulberry extract in the preparation of a product for inhibiting sebum secretion and lipid accumulation in sebaceous glands; preferably, the lipid accumulation includes lipid accumulation in sebaceous gland tissue and / or sebaceous gland cells; preferably, the lipid includes triglycerides (TG) and / or free fatty acids (FFA); (b3) Use of mulberry extract in preparing products for regulating sex hormone levels in sebaceous gland tissue; preferably, any one or more of the following (i)-(iii): (i) use of mulberry extract in preparing products for reducing testosterone (T) and dihydrotestosterone (DHT) content in sebaceous gland tissue; (ii) use of mulberry extract in preparing products for reducing free testosterone index (FAI) in sebaceous gland tissue; (iii) use of mulberry extract in preparing products for increasing estradiol (E2) content in sebaceous gland tissue; (b4) Use of mulberry extract in the preparation of a product for inhibiting abnormal keratinization of hair follicle ducts; (b5) Use of mulberry extract in the preparation of a product for inhibiting excessive proliferation of Propionibacterium acnes; (b6) Use of mulberry extract in the preparation of a product for inhibiting the inflammatory response of sebaceous gland tissue. Preferably, the inhibiting the inflammatory response of sebaceous gland tissue comprises regulating the expression of inflammatory response-related proteins in serum and / or sebaceous gland tissue and / or increasing the antioxidant level in serum and / or sebaceous gland tissue; preferably, comprises reducing the expression of MMP-2 and / or MMP-9 in serum and / or sebaceous gland tissue, and / or increasing the activity of SOD in serum and / or sebaceous gland tissue.

4. The use according to any one of claims 1 to 3, characterized in that: The acne is caused by at least one of excessive sebum secretion from sebaceous glands, abnormal keratinization of the hair follicle duct opening, excessive proliferation of Propionibacterium acnes around the sebaceous gland tissue and / or inflammatory response of the sebaceous gland tissue; preferably, the acne is caused by excessive sebum secretion from sebaceous glands; preferably, the acne is caused by excessive androgen secretion.

5. The use according to any one of claims 1 to 4, characterized in that: The acne is located on the face, neck, chest and / or back where the sebaceous glands are abundant; preferably on the face.

6. The use according to any one of claims 1 to 5, characterized in that: Taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is: Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is: Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is: Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is: Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is: Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is: Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is: Preferably, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is: More preferably, based on the sum of the weight percentage of the components of the mulberry extract being 100%, the weight content of the components in the mulberry extract is: More preferably, based on the sum of the weight percentage of the components of the mulberry extract being 100%, the weight content of the components in the mulberry extract is: Further preferably, the alkaloids include 1-deoxynojirimycin (DNJ), N-methyl-1-deoxynojirimycin (N-methly-1-deoxynojirimycin), fagomine (FAG), 3-epi-fagomine, 1,4-dideoxy-1,4-imino-D-arabinitol (DAB), calystegin B2, calystegin C1, 2-oxo-(α- One or more of 2-O-(α-D-galactopyranosyl)-1-deoxynojirimycin, 6-O-(β-D-glucopyranosyl)-1-deoxynojirimycin, and 1,4-dideoxy-1,4-imino-(2-O-β-D-glucopyranosyl)-D-arabinitol; Preferably, the weight percentage of DNJ is not less than 50% (optionally 60-99%) of the total alkaloids.

7. The use according to any one of claims 1 to 6, characterized in that: The preparation method of the mulberry extract comprises the following steps: 1) preparing a crude extract of a Moraceae plant; 2) separating the crude extract through a cationic resin and / or an optional anionic resin to obtain the mulberry extract; Preferably, the method further comprises the following steps: 3) performing alcohol precipitation on the resin effluent of step 2) and collecting the supernatant; 4) concentrating and / or drying the supernatant; Optionally, the method further comprises the following step: concentrating and / or drying the resin effluent from step 2).

8. The use according to any one of claims 1 to 7, characterized in that: The mulberry extract is mulberry extract, mulberry branch extract, mulberry bark extract and / or mulberry leaf extract.

9. The use according to any one of claims 1 to 8, characterized in that: The product for preventing and / or treating acne is selected from at least one of food, medicine, beverage, health product or cosmetics; preferably, the product is a functional additive.

10. The use according to any one of claims 1 to 9, characterized in that: The mulberry extract acts on humans or mammals.

11. The use according to claim 9, characterized in that: The product is in oral dosage form or external dosage form.

12. The use according to claim 11, characterized in that: The oral dosage forms include tablets, capsules, oral solutions, oral emulsions, pills, granules, syrups or powders.

13. The use according to claim 11, characterized in that: The external dosage forms include lotions, ointments, creams, gels, tinctures, liniments, spirits, powders, oils, pastes, plasters, coatings, and aerosols; preferably, gels.

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