Shampoo with effects of inhibiting bacteria, removing dandruff and relieving itching and preparation method thereof

Abstract

The invention belongs to the technical field of biology. The invention relates to shampoo, in particular to shampoo with antibacterial, dandruff-removing and itching-relieving effects and a preparation method thereof. The shampoo is prepared from the following components in percentage by mass: 5 to 25 percent of surfactant, 0.1 to 0.3 percent of guar gum hydroxypropyl trimethoxy ammonium chloride, 0.3 to 0.5 percent of polyquaternium-10, 0.1 to 0.3 percent of sodium chloride, 0.1 to 0.5 percent of PPG-3 octyl ether, 1 to 3 percent of alpinia katsumadai extract, 0.1 to 1.0 percent of piroctone olamine salt, 2 to 4 percent of humectant, 0.1 to 0.5 percent of chelating agent, 0.01 to 0.05 percent of pH regulator, 0.1 to 0.5 percent of essence and the balance of water. A human body use evaluation test result shows that the shampoo disclosed by the invention has a more remarkable effect on conditions such as red and swollen scalp, scurf and itching in the head compared with a market anti-dandruff shampoo, and the scurf and itching in the head are remarkably improved after a tested patient uses the shampoo for 28 days.

Description

Technical Field

[0001] The present invention belongs to the field of biotechnology, and more specifically, relates to a shampoo with antibacterial, anti-dandruff and antipruritic effects and a preparation method thereof. Background Art

[0002] Seborrheic dermatitis (SD) is a chronic, papular, scaly skin disease with vigorous sebum secretion and Malassezia infection. SD is common on the head, face, chest, and back. When it occurs on the scalp, it is called scalp seborrheic dermatitis / dandruff (SSD). Seborrheic dermatitis is a chronic, recurrent inflammatory scalp disease characterized by the formation of erythema or mild to moderate scales, causing greasy scalp and flaky dandruff accompanied by itching.

[0003] The external pathogens of SSD are mainly the abnormal colonization of Malassezia, whose occupational changes lead to the disorder of the scalp microbial ecosystem. The enzymes and corresponding metabolites produced by Malassezia can directly damage the stratum corneum of the skin, activate the scalp immune mechanism, and produce chronic inflammation. [1] Its mechanism may be related to the activation of pattern recognition receptors, inflammasomes and NF-κB signaling pathways; Malassezia can also block the effect of drugs by forming biofilms, thereby producing drug resistance. [2] Therefore, effectively inhibiting the reproduction of Malassezia in clinical practice is one of the key points in treating this disease. However, in the process of clinical treatment, chemical drugs have obvious side effects and drug resistance, especially the repeated and long-term use of antifungal drugs, which makes Malassezia lack sensitivity to them, often leading to treatment failure.

[0004] 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 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. Studies have found that the volatile oil of Alpinia katsumadai seeds has a certain inhibitory effect on Escherichia coli, Staphylococcus aureus and Bacillus subtilis, but there is no report on the inhibitory effect of Alpinia katsumadai extract on Malassezia.

[0005] [1]Tao R,Li R,Wang R.Skin microbiome alterations in seborrheic dermatitis and dandruff:Asystematic review[J].Exp Dermatol,2021,30(10):1546-1553.

[0006] [2]Adalsteinsson JA, Kaushik S, Muzumdar S, et al. An update on the microbiology, immunology and genetics of seborrheic dermatitis[J]. Experimentaldermatology, 2020, 29(5):481-489. Summary of the invention

[0007] The present invention firstly finds that the cardamom extract has the effect of inhibiting Malassezia, especially Malassezia globosa, and further experiments find that the cardamom extract and the traditional antibacterial agent piroctone ethanolamine salt have a satisfactory synergistic effect, thus obtaining the present invention.

[0008] The invention aims to provide a shampoo with antibacterial, anti-dandruff and antipruritic effects, comprising the following components in percentage by mass: 5-25% of surfactant, 0.1-0.3% of guar hydroxypropyltrimethoxyammonium chloride, 0.3-0.5% of polyquaternium-10, 0.1-0.3% of sodium chloride, 0.1-0.5% of PPG-3 octyl ether, 1-3% of cardamom extract, 0.1-1.0% of piroctone olamine salt, 2-4% of moisturizer, 0.1-0.5% of chelating agent, 0.01-0.05% of pH regulator, 0.1-0.5% of essence and the remainder of water.

[0009] Furthermore, the cardamom extract is the n-hexane extraction part of the cardamom alcohol extract.

[0010] Furthermore, the preparation method of the Alpinia oxyphylla extract is:

[0011] S1. Take dried cardamom seeds, crush them, sieve them, add ethanol, heat and reflux them for extraction, collect the extract, filter, concentrate the filtrate, recover the ethanol, and dry them to obtain a dry extract;

[0012] S2. Add n-hexane to the above dry extract for extraction, concentrate under reduced pressure to constant weight, and dry to obtain.

[0013] Furthermore, in step S1, the heating reflux extraction conditions are: solid-liquid ratio 1:10-30, reaction temperature 60-80°C; reaction time 1-3h; ethanol volume fraction 60-85%.

[0014] Furthermore, in step S2, the parameters of the n-hexane extraction process are: temperature 40-70° C.; extraction time 1.0-2.5 h; and solid-liquid ratio 1:5-10.

[0015] Furthermore, the surfactant is selected from one or a combination of two or more of sodium laureth sulfate, sodium cocoyl methyl taurate, cocamidopropyl betaine, cocamide MEA, sodium cocoyl glycinate, sodium lauroyl sarcosinate, sodium cocoyl glutamate and sodium myristoyl glutamate.

[0016] Furthermore, the moisturizing agent is selected from one or a combination of two or more of allantoin, propylene glycol, sodium hyaluronate, panthenol, glycerin and keratin hydrolyzate.

[0017] Furthermore, the chelating agent is selected from one or a combination of two or more of EDTA, disodium EDTA and tetrasodium glutamate diacetate; and the pH regulator is citric acid.

[0018] Another object of the present invention is to provide a shampoo with antibacterial, anti-dandruff and antipruritic effects, comprising the following components in percentage by mass:

[0019] Sodium laureth sulfate 5-10%;

[0020] Sodium cocoyl methyl taurate 1-5%;

[0021] Cocamidopropyl betaine 1-5%;

[0022] Cocoamide MEA 0.1~1%;

[0023] Guar gum hydroxypropyltrimethoxyammonium chloride 0.1-0.3%;

[0024] Polyquaternium salt - 100.3~0.5%;

[0025] Sodium chloride 0.1~0.3%;

[0026] PPG-3 octyl ether 0.1-0.5%;

[0027] Cardamom extract 1-3%;

[0028] Piroctone olamine salt 0.1-1.0%;

[0029] Panthenol 2-4%;

[0030] Disodium EDTA 0.1-0.5%;

[0031] Citric acid 0.01~0.05%;

[0032] Fragrance 0.1-0.5%; and

[0033] Remaining amount of water.

[0034] Another object of the present invention is to provide a method for preparing the shampoo, comprising the following steps:

[0035] After pre-dispersing guar gum hydroxypropyl trimethoxyammonium chloride in cold water, add disodium EDTA, stir evenly, and heat to 50-70°C; add sodium laureth sulfate, sodium cocoyl methyl taurate, cocamidopropyl betaine and cocamide MEA in sequence, keep warm for 20 minutes, add piroctone ethanolamine salt and stir evenly; after cooling to 30-45°C, add polyquaternium-10 and PPG-3 octyl ether and stir evenly, then add cardamom extract and stir for 30 minutes; cool to room temperature, add panthenol and essence, stir for 30-40 minutes, add citric acid and sodium chloride respectively, and the finished product is discharged.

[0036] The present invention has the following beneficial effects:

[0037] (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.

[0038] (2) The results of the combined drug sensitivity test showed that the combination of the cardamom extract of the present invention and the 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, but showed no effect when used in combination with ZPT and HD-100.

[0039] (3) The results of the human evaluation test showed that the shampoo of the present invention has a more significant effect on dandruff and scalp itching than the anti-dandruff shampoo on the market. After the test patients used it for 28 days, the dandruff and scalp itching phenomena were significantly improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 The dandruff changes of patient No. 10 before and after 28 days of use of the shampoo of Example 1;

[0041] Figure 2 The average changes in scalp oiliness of the two groups of patients before and after 28 days of use;

[0042] Compared with the control group, * P<0.05. DETAILED DESCRIPTION

[0043] 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.

[0044] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0045] 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.

[0046] Examples 1 to 3, antibacterial anti-dandruff shampoo (mass fraction %)

[0047]

[0048]

[0049] Preparation method:

[0050] After pre-dispersing guar gum hydroxypropyl trimethoxyammonium chloride in cold water, add disodium EDTA, stir evenly, and heat to 70°C; add sodium laureth sulfate, sodium cocoyl methyl taurate, cocamidopropyl betaine and cocamide MEA in sequence, keep warm for 20 minutes, add piroctone ethanolamine salt and stir evenly; after cooling to 40°C, add polyquaternium-10 and PPG-3 octyl ether and stir evenly, then add cardamom extract and stir for 30 minutes; cool to room temperature, add panthenol and essence, stir for 30 minutes, add citric acid and sodium chloride respectively, and the finished product is discharged.

[0051] Test Example 1: Comparison of antibacterial activity of different extraction parts of cardamom

[0052] 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.

[0053] 1.2 Test methods

[0054] 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 1×106 CFU / mL of bacterial solution was used as the experimental bacterial suspension.

[0055] 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.

[0056] 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.

[0057] 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.

[0058] 2 Results and discussion

[0059] 2.1 Inhibition zone test results

[0060] 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.

[0061] Table 1 Comparison of the diameters of the inhibition zones of different extraction parts against Malassezia sphaeroides

[0062] 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 <![CDATA[- *** ]]>

[0063] Compared with the n-hexane fraction of cardamom, ** P<0.01, *** P<0.001.

[0064] 2.2MIC test results

[0065] Table 2 MIC test results of n-hexane fraction of cardamom against Malassezia sphaerocephala

[0066] Liquid MIC (μg / mL) Cardamom Hexane 125 OCT 62.5

[0067] Test Example 2: Combined drug sensitivity test

[0068] 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.

[0069] The drug solution was diluted 10 times by double dilution 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°C for 48 hours 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 cypress phenol and other drugs. The calculation is shown in the following formula.

[0070]

[0071] 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.

[0072] Table 3 MIC values ​​of different drugs against Malassezia sphaeroides

[0073] Liquid MIC (μg / mL) Alpinia officinalis Extract 125 OCT 62.5 Ketoconazole 3.91 ZPT 15.63 Climbazole 7.81 Diisothionate 31.25

[0074] Table 4 Test results of combined drug sensitivity test

[0075]

[0076] 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.

[0077] Experiment 3: Human Trial Evaluation

[0078] 1.1 Inclusion criteria: 22 patients with seborrheic dermatitis of the scalp were selected, with an average age of 30±1 years, and randomly divided into 2 groups, with 11 patients in each group. Each patient had moderate or above scalp itching, dandruff, scalp redness and swelling for at least 3 months; hair length was ≥3cm, no severe hair loss, hair root growth covered at least 70% of the scalp; and was healthy and free of disease.

[0079] 1.2 Test method: Use the shampoo of Example 1 and the control group (commercially available Head & Shoulders Icy Cool Anti-dandruff and Antipruritic Shampoo, Guangdong G Makeup Network No. 2022100003), 3 times a week, massage the scalp for at least 1 minute each time and rinse it off, and use it continuously for 28 days. Before the test and on the 28th day, evaluate the degree of dandruff, scalp redness and swelling, and scalp oiliness of the patients, which are recorded as T0D and T28D, respectively.

[0080] Evaluation method: Use scalp image analyzer to observe the changes of dandruff; use skin sebum tester SebumScale to detect oil content; patients use 0-5 points to evaluate the degree of scalp redness and itching. The results are as follows: Figures 1-2 And as shown in Table 5.

[0081] 2 Results

[0082] Table 5 Scalp redness and swelling, scalp itching degree scores

[0083]

[0084] Compared with the control group, * P < 0.05;** P<0.01.

[0085] from Figures 1-2 It can be seen from Table 5 that although the shampoo of the present invention and the commercially available Head & Shoulders Icy Cool Anti-Dandruff and Antipruritic Shampoo both contain piroctone ethanolamine salt, the shampoo of the present invention can better improve the itching, redness, swelling, dandruff and oiliness of the scalp in patients with seborrheic dermatitis.

[0086] 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. A shampoo with antibacterial, anti-dandruff and antipruritic effects, characterized in that: The invention comprises the following components in percentage by mass: 5-25% of surfactant, 0.1-0.3% of guar gum hydroxypropyltrimethoxyammonium chloride, 0.3-0.5% of polyquaternium-10, 0.1-0.3% of sodium chloride, 0.1-0.5% of PPG-3 octyl ether, 1-3% of cardamom extract, 0.1-1.0% of piroctone olamine salt, 2-4% of moisturizing agent, 0.1-0.5% of chelating agent, 0.01-0.05% of pH adjusting agent, 0.1-0.5% of essence and the remainder of water.

2. The shampoo according to claim 1, characterized in that The cardamom extract is the n-hexane extraction part of the cardamom alcohol extract.

3. The shampoo according to claim 2, characterized in that The preparation method of the cardamom extract is: S1. Take dried cardamom seeds, crush them, sieve them, add ethanol, heat and reflux them for extraction, collect the extract, filter, concentrate the filtrate, recover the ethanol, and dry them to obtain a dry extract; S2. Add n-hexane to the above dry extract for extraction, concentrate under reduced pressure to constant weight, and dry to obtain.

4. The shampoo according to claim 3, characterized in that In the step S1, the heating reflux extraction conditions are: solid-liquid ratio 1:10-30, reaction temperature 60-80°C; reaction time 1-3h; ethanol volume fraction 60-85%.

5. The shampoo according to claim 3, characterized in that In step S2, the parameters of the n-hexane extraction process are: temperature 40-70° C.; extraction time 1.0-2.5 h; and solid-liquid ratio 1:5-10.

6. The shampoo according to any one of claims 1 to 5, characterized in that The surfactant is selected from one or a combination of two or more of sodium laureth sulfate, sodium cocoyl methyl taurate, cocamidopropyl betaine, cocamide MEA, sodium cocoyl glycinate, sodium lauroyl sarcosinate, sodium cocoyl glutamate and sodium myristoyl glutamate.

7. The shampoo according to any one of claims 1 to 5, characterized in that The moisturizing agent is selected from one or a combination of two or more of allantoin, propylene glycol, sodium hyaluronate, panthenol, glycerin and keratin hydrolyzate.

8. The shampoo according to any one of claims 1 to 5, characterized in that The chelating agent is selected from one or a combination of two or more of EDTA, disodium EDTA and tetrasodium glutamate diacetate; and the pH regulator is citric acid.

9. A shampoo with antibacterial, anti-dandruff and antipruritic effects, characterized in that: Contains the following components in percentage by weight: Sodium laureth sulfate 5-10%; Sodium cocoyl methyl taurate 1-5%; Cocamidopropyl betaine 1-5%; Cocoamide MEA 0.1~1%; Guar gum hydroxypropyltrimethoxyammonium chloride 0.1-0.3%; Polyquaternium salt - 100.3~0.5%; Sodium chloride 0.1~0.3%; PPG-3 octyl ether 0.1-0.5%; Cardamom extract 1-3%; Piroctone olamine salt 0.1-1.0%; Panthenol 2-4%; Disodium EDTA 0.1-0.5%; Citric acid 0.01~0.05%; Fragrance 0.1-0.5%; and Remaining amount of water.

10. A method for preparing the shampoo according to claim 9, characterized in that: The following steps are involved: After pre-dispersing guar gum hydroxypropyl trimethoxyammonium chloride in cold water, add disodium EDTA, stir evenly, and heat to 50-70°C; add sodium laureth sulfate, sodium cocoyl methyl taurate, cocamidopropyl betaine and cocamide MEA in sequence, keep warm for 20 minutes, add piroctone ethanolamine salt and stir evenly; after cooling to 30-45°C, add polyquaternium-10 and PPG-3 octyl ether and stir evenly, then add cardamom extract and stir for 30 minutes; cool to room temperature, add panthenol and essence, stir for 30-40 minutes, add citric acid and sodium chloride respectively, and the finished product is discharged.

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