Application of cajanin to treatment of acne

Abstract

The invention provides an application of cajanin in treatment of acne. Specifically, the inventor discovers the effect of the cajanin in the aspect of killing the propionibacterium acnes, the cajanin has the effect of killing the propionibacterium acnes under the condition that the concentration of the cajanin is very low, and it is proved that the cajanin has high activity of killing the propionibacterium acnes and has application prospects in the aspects of treating acnes and removing the acnes.

Description

Technical Field

[0001] The invention relates to the field of skin medicine, and in particular to the application of cajanin in treating acne. Background Art

[0002] Acne is one of the most common skin diseases worldwide, specifically targeting the sebaceous follicles on the face, chest and back, affecting most people, especially adolescents. The occurrence of acne involves multiple factors, including hormonal stimulation, increased sebum levels and bacterial colonization. From a microbiological perspective, Propionibacterium acnes can cause acne when it overgrows. Antimicrobial drugs including clindamycin, minocycline and doxycycline are standard treatments for acne, however, the number of antibiotic-resistant Propionibacterium acnes strains has gradually increased over the past decade. Therefore, there is an urgent need to develop new methods for treating acne in this field.

[0003] Cajanus cajan (L.) Millsp. belongs to the genus Cajanus DC. of the family Leguminosae. In addition to its valuable applications in the food and agricultural industries, Cajanus cajan can also be used for medical treatment. Traditionally, the seeds, leaves and roots of Cajanus cajan and their extracts have been used to treat diabetes, jaundice, measles, anti-fungal, avascular necrosis of the femoral head, osteoporosis and antioxidant. In particular, Cajanin, an important active ingredient isolated from Cajanus cajan leaves, can be antioxidant, protect against deoxyribonucleic acid (DNA) damage, relax renal arteries and fight tumors. However, there is currently no application of Cajanin in the treatment of acne. Summary of the invention

[0004] The purpose of the present invention is to provide an application of cajanin in treating acne.

[0005] In the first aspect of the present invention, there is provided a use of pigeon argan oil for

[0006] (1) Inhibit or kill Propionibacterium acnes;

[0007] (2) preparing a medicament for treating and / or preventing acne; and / or

[0008] (3) Preparing cosmetics for anti-acne.

[0009] In another preferred embodiment, the cajanin is obtained by a method selected from the following group: extracted and separated from pigeon pea (Cajanus cajan (L.) Millsp.) or other plants containing cajanin; obtained by chemical synthesis.

[0010] In another preferred embodiment, the concentration of cajanin used to inhibit or kill Propionibacterium acnes is > 4 μg / mL.

[0011] In another preferred embodiment, the concentration is ≥6 μg / mL.

[0012] In another preferred embodiment, the concentration is ≥8 μg / mL.

[0013] In another preferred embodiment, the drug is a pharmaceutical composition, and the pharmaceutical composition contains:

[0014] (1) Active ingredient: caesalpinia; and

[0015] (2) Pharmaceutically acceptable carriers or excipients.

[0016] In another preferred embodiment, the concentration of cajanin in the pharmaceutical composition is >4 μg / mL.

[0017] In another preferred embodiment, the concentration of cajanin in the pharmaceutical composition is ≥6 μg / mL.

[0018] In another preferred embodiment, the concentration of cajanin in the pharmaceutical composition is ≥8 μg / mL.

[0019] In another preferred embodiment, the cosmetic is a cosmetic composition, and the cosmetic composition contains:

[0020] (1) Active ingredient: caesalpinia; and

[0021] (2) A cosmetically acceptable carrier or excipient.

[0022] In another preferred embodiment, the cosmetic composition further contains a second active ingredient, and the second active ingredient is selected from the following group: vitamin A, vitamin C, vitamin E, collagen peptide, hyaluronic acid, apigenin, chlorogenic acid, ferulic acid, kojic acid, niacinamide, arbutin, or a combination thereof.

[0023] In another preferred embodiment, the cosmetically acceptable carrier or excipient is selected from the following group: moisturizers, antioxidants, UV inhibitors, preservatives, film formers, oil-soluble gelling agents, organic modified clay minerals, resins, antibacterial agents, flavors, salts, antioxidants, pH regulators, chelating agents, cooling agents, anti-inflammatory agents, skin beautifying ingredients, vitamins, amino acids, nucleic acids, hormones, inclusion compounds or combinations thereof.

[0024] In another preferred embodiment, the pharmaceutical composition or cosmetic composition is one or more of an emulsion, a liquid, an ointment, a cream, a paste, a cake, and a powder.

[0025] In the second aspect of the present invention, a method for killing Propionibacterium acnes is provided, which comprises administering cajuna or a drug containing cajuna.

[0026] In another preferred embodiment, the method is an in vitro method.

[0027] In another preferred embodiment, the method is non-diagnostic and non-therapeutic.

[0028] In another preferred embodiment, the concentration of cajanin administered in the method is >4 μg / mL.

[0029] In another preferred embodiment, the concentration of cajanin administered in the method is ≥6 μg / mL.

[0030] In another preferred embodiment, the concentration of cajanin administered in the method is ≥8 μg / mL.

[0031] In a third aspect of the present invention, a method for treating and / or preventing acne is provided, wherein cajuna or a medicament containing cajuna is administered to a subject.

[0032] In another preferred embodiment, the subject is a human.

[0033] In another preferred embodiment, the subject suffers from acne.

[0034] In another preferred embodiment, the subject is a person prone to acne.

[0035] In a fourth aspect of the present invention, a cosmetic method is provided, wherein the cosmetic comprises administering caesalpinia or a cosmetic containing caesalpinia to a subject.

[0036] In another preferred embodiment, the subject is a human.

[0037] In another preferred embodiment, the subject suffers from acne.

[0038] In another preferred embodiment, the subject is a person prone to acne.

[0039] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 The results show that the 10 dilution after 48 h of liquid culture -2 (left) and 10 -4 (Right) Plate counting results and colony morphology at 72h.

[0041] Figure 2 The figure shows the culture medium for 48 hours of liquid culture. The left picture shows the culture medium in a static state, the culture medium is clear and transparent, and white tea stain-like precipitation can be seen at the bottom; the right picture shows that the culture medium is a semi-permeable emulsion after shaking.

[0042] Figure 3 Shown are 96-well microplate cultures after incubation with P. acnes and different concentrations of Cajunin for 48 hours.

[0043] Figure 4 The results show the inhibitory effect of different concentrations of caesarin on Propionibacterium acnes. The results show that caesarin can effectively kill Propionibacterium acnes at 8μg / mL. In the figure, A shows the plate culture results of the control group without drug addition; the upper figure of B shows the plate culture results when the drug concentration is 8μg / mL, and the lower figure shows the plate culture results when the drug concentration is 4μg / mL. DETAILED DESCRIPTION

[0044] After extensive and in-depth research, the inventors discovered for the first time the effect of caesin on killing Propionibacterium acnes. By measuring the minimum bactericidal concentration experimentally, it was found that caesin had the effect of killing Propionibacterium acnes at a very low concentration, proving that caesin has a strong activity of killing Propionibacterium acnes and has application prospects in the treatment of acne. On this basis, the present invention was completed.

[0045] Propionibacterium acnes

[0046] Propionibacterium acnes is a Gram-positive anaerobic or facultative anaerobic pathogen that causes acne.

[0047] P. acnes is usually a commensal of sebaceous glands and can cause acne when it grows excessively. The main virulence factor of P. acnes is secretory lipase, which acts on triglycerides to release free fatty acids. Among them, palmitic acid stimulates Toll-like receptor 2-mediated inflammasomes, which is associated with the release of interleukin 1, Th17 differentiation, and interleukin 17-mediated keratinocyte proliferation; while oleic acid also stimulates the adhesion of P. acnes, keratinocyte proliferation, and acne occurrence through the release of interleukin 1.

[0048] pigeondarin

[0049] Pigeon pea is an active ingredient isolated from pigeon pea leaves. Its chemical structure is shown below:

[0050]

[0051] The pigeon pea extract mentioned in the present invention can be derived from pigeon pea extract. The pigeon pea extract contains pigeon pea extract. The pigeon pea extract can be obtained by conventional methods in the art such as extraction, concentration, extraction, silica gel column chromatography, gel column separation, etc.

[0052] In some embodiments, the pigeon arganin is from a pigeon arganin-containing extract.

[0053] In some embodiments, the pigeon pea extract is extracted from the root, stem, leaf, or fruit of pigeon pea, preferably, the leaf.

[0054] In some embodiments, the pigeon pea is produced in Yunnan, Sichuan, Jiangxi, Hunan, Guangxi, Guangdong, Hainan, Zhejiang, Fujian, Taiwan, Jiangsu, preferably, Yunnan.

[0055] In some embodiments, the cajanin is artificially synthesized.

[0056] Pigeon argan extract kills Propionibacterium acnes

[0057] In the present invention, the inhibitory ability of caesarin on Propionibacterium acnes was determined experimentally, the minimum bactericidal concentration (MBC) of caesarin on Propionibacterium acnes was measured, and the minimum inhibitory concentration (MIC) was inferred.

[0058] Minimum bactericidal concentration (MBC) refers to the minimum concentration that can kill bacteria in the culture medium (i.e., kill 99.9% of the test microorganisms).

[0059] Minimum inhibitory concentration (MIC) refers to the minimum drug concentration that can inhibit the growth of pathogens in the culture medium. Generally, MBC is greater than or approximately equal to MIC.

[0060] In order to determine the inhibitory ability of caesarin on Propionibacterium acnes, the minimum bactericidal concentration of caesarin on Propionibacterium acnes was measured as follows: equal amounts of Propionibacterium acnes were incubated with different concentrations of caesarin, and the minimum bactericidal concentration was obtained when the killing level of Propionibacterium acnes reached 99.9%. It is inferred that the minimum inhibitory concentration is less than the minimum bactericidal concentration.

[0061] The experiment of the present invention proves that the minimum bactericidal concentration of caesarin against Propionibacterium acnes is 8 μg / mL, and the minimum inhibitory concentration of caesarin against Propionibacterium acnes is less than 8 μg / mL and should be greater than 4 μg / mL, indicating that caesarin has a strong killing effect on Propionibacterium acnes.

[0062] application

[0063] The inventors found that pigeon argan has the function of killing Propionibacterium acnes and can be used to prepare a pharmaceutical composition for treating acne.

[0064] Pigeonol can be used to prepare pharmaceutical compositions, such as tablets, capsules, powders, microgranules, solutions, lozenges, jellies, cream preparations, spirits, suspensions, tinctures, poultices, liniments, lotions, and aerosols. The medicine can be prepared by commonly known preparation techniques, and suitable pharmaceutical additives can be added to the medicine.

[0065] The pharmaceutical composition of the present invention contains a safe and effective amount of the active ingredient of the present invention and a pharmaceutically acceptable carrier. Such carriers include (but are not limited to): saline, buffer, glucose, water, glycerol, ethanol, and combinations thereof. Usually, the pharmaceutical preparation should match the mode of administration. The dosage form of the pharmaceutical composition of the present invention is injection, oral preparation (tablet, capsule, oral liquid), transdermal agent, sustained release agent. For example, it is prepared by conventional methods using physiological saline or an aqueous solution containing glucose and other adjuvants. The pharmaceutical composition is preferably manufactured under sterile conditions.

[0066] In addition, caesalpinol can also be used to prepare cosmetic compositions for removing acne. The present invention also provides a cosmetic composition containing caesalpinol.

[0067] The cosmetic composition of the present invention comprises:

[0068] (a) cajanin as the first active ingredient; and / or

[0069] (b) other ingredients with skin care effects selected from the group consisting of vitamin A, vitamin C, vitamin E, collagen peptides, hyaluronic acid, apigenin, chlorogenic acid, ferulic acid, kojic acid, niacinamide, arbutin, or a combination thereof; and

[0070] (c) a pharmaceutically acceptable carrier or excipient.

[0071] The main advantages of the present invention include:

[0072] (1) The present invention provides for the first time a new use of cajanin in treating acne and / or removing acne.

[0073] (2) The present invention found that even at very low concentrations, pigeon argan oil has an excellent effect in killing Propionibacterium acnes.

[0074] (3) The present invention provides for the first time a drug for treating acne and / or removing acne containing caesalpinol.

[0075] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended only to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods for the following examples without specifying specific conditions are usually based on the conditions described in the normal conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and weight parts.

[0076] Example 1: Propionibacterium acnes and its cultivation method

[0077] Culture conditions: 36℃-37℃, anaerobic.

[0078] The solid culture medium used for the culture is a nutrient agar medium, which contains: 10 g of tryptone, 10 g of beef extract, 5 g of glucose, 5 g of sodium chloride, 3 g of yeast extract, 3 g of sodium acetate, 1 g of soluble starch, 0.5 g of L-cysteine ​​hydrochloride, and 15 g of agar; pH value is 6.8. The liquid culture medium is a nutrient agar medium without agar.

[0079] Growth rate: Visible colonies appear on nutrient agar medium after 48 hours, and clearly visible colonies appear after 72 hours.

[0080] Culture results: The colony morphology is milky yellow, translucent, rounded protrusions ( Figure 1 ); the culture medium was statically cultured in liquid culture medium, the culture medium was clear and transparent, white tea stain-like precipitation was visible at the bottom, and after shaking, the culture medium was semi-permeable emulsion ( Figure 2 ), cultured for 48-72h, colony concentration (>10 7 cfu / mL).

[0081] Example 2: Obtaining pigeon argan

[0082] 2.1 Obtaining Pigeon Pea Extract from Pigeon Pea Leaves

[0083] Pigeon pea source: Honghe County Taifu Agriculture and Forestry Technology Development Co., Ltd.;

[0084] Product name: Medicinal pigeon pea;

[0085] Origin: Honghe County, Yunnan Province.

[0086] (1) Extraction

[0087] Take 100 g of dried pigeon pea leaves and cut them into pieces. Add 6 times the amount of 80% ethanol-water solution (based on the total volume of the solution) by mass volume ratio, reflux extraction for 1 hour, filter, and collect the filtrate. Repeat the extraction three times.

[0088] (2) Concentration

[0089] Concentrate the filtrate below 50°C until there is no alcohol taste.

[0090] (3) Extraction

[0091] The residue after concentration is suspended in water, extracted three times with an equal volume of petroleum ether, and the petroleum ether layer is concentrated to obtain 20-25 g of petroleum ether extract (pigeon pea extract).

[0092] 2.2 Preparation of pigeon argan

[0093] (1) Sample mixing

[0094] The petroleum ether extract (pigeon pea extract) prepared in Example 1 was dissolved in an appropriate amount of ethyl acetate, and an appropriate amount of 100-200 mesh silica gel was added, and dried under reduced pressure.

[0095] (2) Silica gel column chromatography

[0096] Soak an appropriate amount of 100-200 mesh silica gel with n-hexane, pack it into the column, with a diameter-to-height ratio of 1:6-10, and load the sample by dry method. First rinse with 2 times the column volume of n-hexane, then change to n-hexane: ethyl acetate = 25-4:1 gradient elution, with each 50 ml as one fraction.

[0097] (3) Thin layer chromatography detection

[0098] The developing solvent was n-hexane:ethyl acetate = 3:1.

[0099] (4) LH-20 gel separation

[0100] Combine the target fractions, concentrate to dryness, dissolve with a small amount of CH2Cl2, and apply to LH-20 gel column. The eluent is petroleum ether: CH2Cl2: methanol = 1:2:1, and HPLC tracking detection is performed.

[0101] HPLC conditions:

[0102] Mobile phase A: 0.1% formic acid water, C: methanol; A:B = 16:84.

[0103] Chromatographic column: 250×4.6nm, 5μm, flow rate 1ml / min, detection wavelength: 302nm.

[0104] The retention time of caesalpin is: 10.5min.

[0105] The purity of 99% of cajanin was isolated.

[0106] Example 3: Determination of the minimum bactericidal concentration (MBC) of pigeon argan against Propionibacterium acnes

[0107] This example determines the minimum bactericidal concentration of pigeon argan, and the method is as follows:

[0108] PBS was prepared with different concentrations of cajanin (μg / mL). Propionibacterium acnes (90 μl; 10 5 CFU / ml (PBS solution) and different concentrations of caenorhain (10 μl; 4–1000 μg / mL in PBS), and only 10 μl PBS was added as a control. Incubate overnight at 30°C.

[0109] MBC was defined as a killing level of 99.9% and was determined by spotting the culture fluid (5 μl) on agar plates supplemented with medium for CFU counting.

[0110] The concentration of cajanin in each well of the 96-well microplate is shown in the following table (μg / mL). K refers to sterile culture medium, C refers to the control without drug addition, and the values ​​represent the final concentration of the drug in each well. The initial concentration of colonies in all wells is about 10 5 cfu / mL.

[0111]

[0112]

[0113] The results showed that after 48 hours, the transparency of the control wells decreased and obvious bacterial film formed on the bottom. All wells with a concentration of caenorhain ≥ 8 μg / mL were transparent and no bacterial film was formed. Bacterial film was visible in the wells with a concentration of 4 μg / mL. Figure 3 shown.

[0114] Take the culture medium of each concentration and transfer it to a plate for culture. Figure 4 As shown, when the concentration is 4 μg / mL, there are colonies on the plate; when the concentration is 8 μg / mL, there are no colonies. The above results prove that the minimum bactericidal concentration (MBC) of caesalpin is 8 μg / mL, and the predicted minimum inhibitory concentration (MIC) is less than 8 μg / mL, proving that caesalpin has a strong activity of killing Propionibacterium acnes.

[0115] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A use of pigeon arone, characterized in that, Used for (1) Inhibit or kill Propionibacterium acnes; (2) preparing a medicament for treating and / or preventing acne; and / or (3) Preparing cosmetics for anti-acne.

2. The use according to claim 1, characterized in that The cajanin is obtained by a method selected from the following group: extracted and separated from pigeon pea (Cajanus cajan (L.) Millsp.) or other plants containing cajanin; obtained by chemical synthesis.

3. The use according to claim 1, characterized in that The concentration of cajanin used to inhibit or kill Propionibacterium acnes was > 4 μg / mL.

4. The use according to claim 1, characterized in that The concentration of cajanin used to inhibit or kill Propionibacterium acnes is ≥ 8 μg / mL.

5. The use according to claim 1, characterized in that The medicine is a pharmaceutical composition, which contains: (1) Active ingredient: caesalpinia; and (2) Pharmaceutically acceptable carriers or excipients.

6. The use according to claim 1, characterized in that The cosmetic is a cosmetic composition, and the cosmetic composition contains: (1) Active ingredient: caesalpinia; and (2) A cosmetically acceptable carrier or excipient.

7. The use according to claim 5 or 6, characterized in that The pharmaceutical composition or cosmetic composition is one or more of an emulsion, a liquid, an ointment, a cream, a paste, a cake, and a powder.

8. A method for killing Propionibacterium acnes, characterized in that: Administer cenobamate or medications containing cenobamate.

9. A method for treating and / or preventing acne, characterized in that: The subject is administered with caesarin or a drug containing caesarin.

10. A cosmetic method, characterized in that: The subjects were administered with caesarin or cosmetics containing caesarin.

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