Scalp care composition with effects of inhibiting bacteria, removing dandruff and relieving itching as well as preparation method and application of scalp care composition

Abstract

The invention belongs to the technical field of biology. The invention relates to the technical field of scalp care, in particular to a scalp care composition with the effects of inhibiting bacteria, removing dandruff and relieving itching as well as a preparation method and application of the scalp care composition, and particularly, the scalp care composition comprises an Alpinia katsumadai extract and piroctone olamine salt. The alpinia katsumadai extract disclosed by the invention has relatively good bacteriostatic activity on malassezia sphaeroides, and a combined drug sensitive test shows that the alpinia katsumadai extract and piroctone olamine salt are combined for use to achieve a synergistic effect in the aspect of inhibiting malassezia sphaeroides.

Description

Technical Field The present invention belongs to the field of biotechnology and more specifically relates to a scalp care composition with antibacterial, anti-dandruff and antipruritic effects and a preparation method and application thereof, in particular, the composition comprises an Alpinia katsu madai extract and piroctone olamine salt. Background Art Malassezia mainly grows in the stratum corneum of human skin and is related to the triglycerides and free fatty acids present in the sebaceous glands on the skin surface. It can cause a variety of skin diseases, such as tinea versicolor, Malassezia folliculitis, seborrheic dermatitis and dandruff. At present, the drugs that inhibit Malassezia are mainly chemical antifungal preparations, such as ketoconazole, climbazole, zinc pyrithione, etc. However, most of them are chemically synthesized compounds. Long-term use of such substances may cause patients to have dry scalp, a lot of desquamation, and are prone to drug resistance. Alpinia katsumadai is the dried nearly mature seeds of the perennial herb Alpinia katsumadai of the Zingiberaceae family. It has the effects of drying dampness, promoting the flow of qi, warming the middle and stopping vomiting. Studies have shown that the volatile oil in Alpinia katsumadai has the effects of protecting gastric mucosa, resisting gastric ulcers, anti-inflammatory, and antibacterial. [1-3] . Wei Jieshu et al. [4] Studies have found that the volatile oil from cardamom seeds has a certain inhibitory effect on Escherichia coli, Staphylococcus aureus and Bacillus subtilis. [5] Studies have shown that cardamom volatile oil can inhibit the pathogenic bacteria of dairy cow mastitis. [6] It was found that the microemulsion of volatile oil of cardamom had a good inhibitory effect on banana anthracnose and had a certain preservation effect on bananas. However, there was no report on the inhibitory effect of cardamom extract on Malassezia.

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[0002] Wu Zhen, Chen Yongshun, Du Shiming, et al. Effects of cardamom volatile oil on acetic acid-induced gastric ulcer in rats[J]. Chinese Journal of Hospital Pharmacy, 2010, 30(7): 560-563.

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[0004] Wei Jieshu, Hu Jinshan, Hong Qunhua, et al. Study on the antibacterial activity of volatile oils from Amomum villosum and Amomum villosum[J]. Biochemical Engineering, 2023, 9(3): 45-50.

[0005] Peng Fu, Dai Min, Wan Feng. Study on the antibacterial activity of the effective parts of Alpinia oxyphylla against the pathogens of bovine mastitis[J]. Chinese Journal of Veterinary Science, 2012, 42(10): 1073-1080.

[0006] Chu Zuochen. Preparation of cardamom essential oil microemulsion and its effect on banana preservation[D]. Wuhan: Huazhong Agricultural University, 2022. Summary of the invention The present invention has found through research that the specific solvent extraction part of the cardamom alcohol extract has a significant inhibitory effect on Malassezia spherica, and the effect is most obvious compared with other extraction parts. Through further research on its interaction with traditional anti-dandruff agents, it is found that the cardamom extract only shows a synergistic effect FICI≤0.5 with piroctone olamine salt, and shows an additive effect when used in combination with ketoconazole and climbazole, and shows an irrelevant effect when used in combination with ZPT and HD-100. The present invention aims to provide use of an extract of Alpinia fasciata in preparing a Malassezia inhibitor. Another object of the present invention is to provide a use of an extract of Alpinia oxyphylla in the preparation of a medicine for treating skin diseases caused by Malassezia infection. In one embodiment of the present invention, the skin diseases include tinea versicolor and seborrheic dermatitis. Another object of the present invention is to provide a use of the cardamom extract in preparing an antibacterial, anti-dandruff and antipruritic scalp care product. In one embodiment of the present invention, the Malassezia is Malassezia globosa. In one embodiment of the present invention, the cardamom extract is the n-hexane extraction portion of the cardamom alcohol extract. Another object of the present invention is to provide an antibacterial, anti-dandruff and antipruritic scalp care composition, which comprises an extract of cardamomum villosum with a concentration of about 25 to 150 μg / mL and piroctone olamine with a mass percentage of about 0.1 to 1.0%. In one embodiment of the present invention, the composition further comprises: a pharmaceutically and / or cosmetically acceptable carrier. In one embodiment of the present invention, the pharmaceutically and / or cosmetically acceptable carrier comprises: a combination of two or more of a surfactant, a humectant, a chelating agent, an emollient, a pH adjuster and a fragrance. In one embodiment of the present invention, the composition is prepared in the form of shampoo, conditioner, hair essence, hair cream or hair gel. The present invention has the following beneficial effects: (1) The results of the inhibition zone test and the MIC value test showed that the diameter of the inhibition zone of the cardamom extract of the present invention against Malassezia sphaerocephala was greater than 20 mm, and the antibacterial effect was the most obvious, which was extremely sensitive. (2) The results of the combined drug sensitivity test showed that the combination of cardamom extract and traditional anti-dandruff agents did not show a synergistic effect. It only showed a synergistic effect when used in combination with OCT, with FICI ≤ 0.5. It showed an additive effect when used in combination with ketoconazole and climbazole, and showed no effect when used in combination with ZPT and HD-100. DETAILED DESCRIPTION The present invention is further described below with reference to specific examples, but the examples do not limit the present invention in any form. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art. Unless otherwise specified, the reagents and materials used in the following examples are commercially available. Among them, the Malassezia solid culture medium: 1% peptone, 2% agar, 1% glucose, 0.2% yeast extract, 0.05% monostearate glyceride, 0.5% Tween-60, 1% glycerol, 0.8% ox bile salt and 2% olive oil, fixed to 1000mL, pH7.0-7.2; the liquid culture medium does not contain agar. Test Example 1: Comparison of antimicrobial activity of different extracts of cardamom 1.1. Preparation of different extraction parts of cardamom: Take dry cardamom seeds, crush them into coarse powder and sieve them, add 65% ethanol solution with a volume fraction of 1:10, soak for 30 minutes, heat and reflux at 70°C for 1.5 hours each time, collect the extract, filter, and remove the residue; concentrate the filtered liquid, refrigerate to precipitate ethanol, recover the precipitated ethanol, dry the supernatant to obtain a dry extract. Add 5 times the weight of n-hexane, petroleum ether, chloroform and ethyl acetate to the above dry extract, extract at 45°C for 2.0 hours, concentrate under reduced pressure to constant weight, dry, and obtain n-hexane, petroleum ether, chloroform and ethyl acetate extraction parts respectively for standby use. 1.2 Test methods 1.2.1 Activation of strains and preparation of bacterial suspension: The test strain (Malassezia globosa CBS9757, purchased from the Medical Fungus Collection Center, Institute of Dermatology, Chinese Academy of Medical Sciences) was subcultured twice on a solid medium of Malassezia, and a single colony was picked and inoculated into a liquid medium of Malassezia, and incubated at 32°C in a shaking incubator at 220 r / min for 48 h. The bacterial suspension was diluted to a final concentration of 1×10 6 CFU / mL of bacterial solution was used as the experimental bacterial suspension. 1.2.2 Preparation of drug solution: The n-hexane, petroleum ether, chloroform and ethyl acetate extraction parts were dissolved in dimethyl sulfoxide to prepare a drug solution with a mass concentration of 100 mg / mL, and stored at 4°C for use; the concentration of the positive drug piroctone ethanolamine (OCT) was 50 mg / mL. 1.2.3 Inhibition zone test: According to the procedure reported by the Clinical and Laboratory Standards Institute (CLSI), the antibacterial activity of each drug solution was tested by the Kirby-Bauer paper diffusion method. Use a pipette to draw 200 μL of bacterial suspension on the culture medium plate, evenly spread it and set aside. Add 20 μL of drug solution to a sterile round filter paper with a specification of (6.0 mm × 1.0 mm). Use tweezers to gently stick the filter paper to the surface of the agar plate. After the plate is placed at 37°C for 48 hours, observe and measure the diameter of the inhibition zone, and take the average value as the measurement result. The edge of the inhibition zone is limited to the naked eye without obvious bacterial growth. The results are determined according to the "Antimicrobial Susceptibility Test Implementation Standard" promulgated by the National Committee for Clinical Laboratory Standards (NCCLS): diameter > 20 mm is extremely sensitive, 15-20 mm is highly sensitive, 10-15 mm is moderately sensitive, and < 10 mm is low sensitive. 1.3 MIC value determination: The reference standard is the M27 standard developed by the Clinical and Laboratory Standards Institute (CLSI) - the reference method for antifungal susceptibility test of yeast broth dilution. Use distilled water to dilute the drug solution 10 times by two-fold dilution method, take a 96-well plate, add 96 μL of liquid culture medium to columns 1 to 11, add 4 μL of a single drug solution of 10 gradients to column 1, and the concentration of the drug solution is from high to low. Add 100 μL of bacterial solution to each well in columns 1 to 11, and make 3 parallels for each drug. Column 11 is used as a growth control, and an OCT positive control group and a negative control group are set. The inoculated 96-well plate is placed at 30°C for 48 hours and the results are observed. The positive well shows a color change from yellow to red. The well with the lowest drug concentration where the fungus is completely inhibited (no color change) is the MIC. 2 Results and discussion 2.1 Inhibition zone test results The results of the inhibition zone are shown in Table 1 below. The diameter of the inhibition zone of the n-hexane part of 50 mg / mL cardamomum against Malassezia globosa is >20 mm, and the antibacterial effect is the most obvious, which is extremely sensitive; the diameter of the inhibition zone of the chloroform part of the cardamomum at the same concentration against Malassezia is <10 mm; the petroleum ether extraction part has a moderately sensitive antibacterial effect on Malassezia, while the ethyl acetate part has no antibacterial activity. Table 1 Comparison of the diameters of the inhibition zones of different extraction parts against Malassezia sphaeroides Liquid Diameter of inhibition zone (mm) Cardamom Hexane 32.62±2.13 Cardamom petroleum ether fraction <![CDATA[13.52±1.03 ** ]]> Cardamom chloroform fraction <![CDATA[8.29±0.75 *** ]]> Cardamom ethyl acetate fraction _*** Compared with the n-hexane fraction of cardamom, ** P<0.01, *** P<0.001. 2.2 MIC test results Table 2 MIC test results of n-hexane fraction of cardamom against Malassezia sphaerocephala Liquid MIC (μg / mL) Cardamom Hexane 125 OCT 62.5 Test Example 2: Combined drug sensitivity test The checkerboard dilution method was used for the combined drug sensitivity test, and the partial inhibitory concentration index (FICI) was determined to evaluate the combined drug effect of the n-hexane fraction of cardamom (hereinafter referred to as cardamom extract) and other drugs. The maximum stock concentration of cardamom extract was 100 mg / mL, the maximum stock concentration of piroctone olamine (OCT) and dihydroxyethyl sulfonate (HD-100) was 50 mg / mL, and the maximum stock concentration of ketoconazole, zinc pyrithione (ZPT) and climbazole was 25 mg / mL. The drug solution was diluted 10 times by double dilution method with distilled water, and 96 μL of liquid culture medium was added to the 1st to 10th columns of the 96-well plate. 2 μL of the multiple concentration solutions of the two drugs were added horizontally and vertically, respectively. There were 10 concentration gradient solutions of cardamom extract from high to low in 10 rows horizontally, and 10 concentration gradient solutions of traditional antidandruff agents from high to low in 10 columns vertically, so that the two drugs formed different combined drug concentrations in 80 wells. Then 100 μL of bacterial solution was added to each well of the 1st to 11th columns. The 11th well was used as a positive control, and the 12th well was used as a negative control. The 96-well plate was placed at 30℃ for 48 h to observe the results. The color of the positive wells changed from yellow to red. The partial inhibitory concentration index (FICI) was used to evaluate the combined effect of cypermethrin and other drugs. The calculation formula is shown in the following formula. Among them, A and B are cardamom extract and traditional anti-dandruff agent, respectively, and MICA and MICB are the MIC values ​​of A and B when used alone, respectively. 联用 is the MIC value of A when A and B are used together, MICB 联用 It is the MIC value of B when A and B are used together. When FICI≤0.5, it is a synergistic effect; when 0.5<FICI≤1, it is an additive effect; when 1<FICI≤2, it is an irrelevant effect; when FICI>2, it is an antagonistic effect. Table 3 MIC values ​​of different drugs against Malassezia sphaeroides Liquid MIC (μg / mL) Alpinia officinalis Extract 125 OCT 62.5 Ketoconazole 3.91 ZPT 15.63 Climbazole 7.81 Diisothiocyanate 31.25 Table 4 Test results of combined drug sensitivity test It can be seen from the above table that the combination of cardamom extract and traditional anti-dandruff agents does not show a synergistic effect. It only shows a synergistic effect when used in combination with OCT, with FICI ≤ 0.5. It shows an additive effect when used in combination with ketoconazole and climbazole, and has no effect when used in combination with ZPT and HD-100. The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. Use of cardamom extract in preparing Malassezia inhibitors.

2. Use of cardamom extract in preparing medicines for treating skin diseases caused by Malassezia infection.

3. The use according to claim 2, characterized in that: The skin diseases include tinea versicolor and seborrheic dermatitis.

4. Use of cardamom extract in preparing antibacterial, anti-dandruff and antipruritic scalp care products.

5. The use according to any one of claims 1 to 4, characterized in that: The Malassezia is a globose Malassezia.

6. The use according to any one of claims 1 to 4, characterized in that: The cardamom extract is the n-hexane extraction part of the cardamom alcohol extract.

7. An antibacterial, anti-dandruff and antipruritic scalp care composition, characterized in that: The invention comprises a cardamom extract with a concentration of about 25 to 150 μg / mL and a piroctone ethanolamine salt with a mass percentage of about 0.1 to 1.0%.

8. The scalp care composition according to claim 7, characterized in that The composition further comprises: a pharmaceutically and / or cosmetically acceptable carrier.

9. The scalp care composition according to claim 8, characterized in that The pharmaceutically and / or cosmetically acceptable carrier comprises: a combination of two or more of a surfactant, a humectant, a chelating agent, an emollient, a pH adjuster and a fragrance.

10. The scalp care composition according to claim 9, characterized in that The composition is prepared in the form of shampoo, hair conditioner, hair essence, hair cream or hair gel.

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