Grapefruit peel shower gel and its application

Abstract

This application provides a shower gel containing grapefruit peel polysaccharides and its use in treating skin diseases such as psoriasis. The shower gel of this application uses grapefruit peel polysaccharides modified with pectinase or polygalacturonase, as well as ingredients such as naringin and naringenin, to provide better therapeutic effects than unmodified grapefruit peel polysaccharides. The active ingredients in this shower gel product are natural ingredients treated with edible enzymes. It can be used frequently and conveniently in daily life, is safe and effective, and provides a novel option for controlling skin problems such as psoriasis.

Description

Technical Field

[0001] The present application belongs to the field of cosmetics and sugar engineering technology. Specifically, the present application provides a shower gel containing grapefruit peel polysaccharide and its application in treating skin diseases such as psoriasis. Background Art

[0002] As a subtropical fruit widely processed and consumed in my country, the consumption of pomelo and the processing of related products generate a large amount of inedible pomelo peel waste. Pomelo peel contains a variety of active substances, including polysaccharides, polyphenols, and flavonoids, such as naringin and naringenin, which can be used in the production of a variety of pharmaceuticals, foods, and health supplements. However, aside from pomelo tea and pastries, the development of other pomelo peel products is currently underdeveloped.

[0003] There is no specific treatment for various difficult-to-cure skin diseases, represented by psoriasis, in both traditional Chinese and Western medicine. Long-term conditioning using plant polysaccharides, polyphenols or peptide products with antioxidant, antibacterial and immune-regulating functions is a good choice for treating and controlling the disease, but the efficacy of such products needs to be improved.

[0004] Enzyme modification is a commonly used method to improve the molecular structure and properties of polysaccharides. Treatment with certain hydrolases can not only reduce the molecular weight, but also change the spatial structure of polysaccharide molecules, such as the triple helix structure, thereby affecting the physiological activity of polysaccharides. Summary of the Invention

[0005] To solve the above problems and further enhance the role of grapefruit peel polysaccharide in treating skin problems, especially psoriasis, the applicant has studied a method for enzymatically modifying grapefruit peel polysaccharide and used it in shower gel.

[0006] In one aspect, the present application provides a grapefruit peel shower gel, comprising grapefruit peel polysaccharide.

[0007] On the other hand, grapefruit peel polysaccharide is used in the preparation of a medicine for treating psoriasis, wherein the medicine is in the form of a shower gel.

[0008] Furthermore, the shower gel further comprises naringin and naringenin ingredients.

[0009] Furthermore, the grapefruit peel polysaccharide is enzyme-modified grapefruit peel polysaccharide.

[0010] Furthermore, the grapefruit peel polysaccharide is grapefruit peel polysaccharide modified by pectinase or polygalacturonase.

[0011] Furthermore, the enzyme-modified grapefruit peel polysaccharide is prepared according to the following method:

[0012] (1) Prepare grapefruit peel polysaccharide at a concentration of 1-10 mg / mL;

[0013] (2) adding pectinase to 50-200 U / mL, pH 5.0-7.0, and treating at 40-60 degrees Celsius for 1-4 hours; or adding polygalacturonase to 50-100 U / mL, pH 5.0-7.0, and treating at 40-60 degrees Celsius for 1-4 hours;

[0014] (3) Inactivate the enzyme; cool, concentrate and freeze-dry.

[0015] Furthermore, in step (1), the concentration of grapefruit peel polysaccharide is prepared to be 5 mg / mL;

[0016] Furthermore, in step (2), pectinase is added to 100 U / mL, pH 6.0, and treated at 40 degrees Celsius for 2 hours; or polygalacturonase is added to 80 U / mL, pH 6.0, and treated at 40 degrees Celsius for 2 hours.

[0017] Furthermore, the shower gel further comprises ingredients selected from surfactants, moisturizers, fragrances, and glossing agents.

[0018] Furthermore, the shower gel is prepared from the following raw materials: 50 parts of deionized water, 10 parts of sodium laureth sulfate, 10 parts of ammonium laureth sulfate, 5 parts of ammonium lauryl sulfate, 5 parts of cocamide, 10 parts of cocamidopropyl betaine, 2 parts of glycerin, 3 parts of sodium lauroyl sarcosinate, 2 parts of acrylic copolymer, 0.2 parts of fragrance, 2 parts of citric acid, 2 parts of glossing agent, and 4 parts of grapefruit peel polysaccharide.

[0019] Furthermore, the grapefruit peel polysaccharide is prepared according to the following method: the grapefruit peel is dried, crushed, and passed through a 100-mesh sieve; deionized water is added according to a material-liquid ratio of 1:30 mass / volume, and the pH is adjusted to 5.0; extraction time is 2 hours at 100 degrees Celsius; after cooling, filtration is performed; 3 times the volume of ethanol is added for precipitation; the precipitate is obtained by centrifugation, concentrated, and freeze-dried.

[0020] In addition, the present application also provides a method for preparing the above-mentioned grapefruit peel polysaccharide.

[0021] The raw material for preparing grapefruit peel polysaccharides in this application is not limited to the pomelo used in the examples; other grapefruit varieties, such as pomelo and Thai pomelo, can also be used. Grapefruit peel polysaccharides can be prepared using known methods or commercially available products. Those skilled in the art can appropriately adjust the extraction and enzyme modification methods based on the specific grapefruit peel polysaccharide.

[0022] The present application may also include other components in grapefruit peel, such as naringin and naringenin, which can be extracted and prepared in the same process as grapefruit peel polysaccharides, or can be extracted separately or purchased commercially; their functions such as anti-inflammatory, immunomodulatory, and antioxidant functions are well known to those skilled in the art.

[0023] The enzymes in this application do not have specific manufacturers or contents. Within the common content range, those skilled in the art can calculate and adjust the dosage accordingly.

[0024] The shower gel of the present application can be used for the treatment or daily conditioning of various skin diseases, including but not limited to psoriasis, dermatitis, acne, etc. Since its active ingredients are natural ingredients treated with edible enzymes, it can also be used for daily use for non-therapeutic purposes. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 Effects of pectinase (A), α-amylase (B), β-amylase (C), polygalacturonase (D), pectinesterase (E), pectin lyase (F), arabinofuranosidase (G), xylanase (H) and untreated grapefruit peel polysaccharide (I) on the survival of HaCat cells, a psoriasis cell model (J is a control with only culture medium added but no polysaccharide added, n=5).

[0026] Figure 2 The DPPH and superoxide anion radical scavenging effects of pectinase (A), α-amylase (B), β-amylase (C), polygalacturonase (D), pectinesterase (E), pectin lyase (F), arabinofuranosidase (G), xylanase (H) and untreated grapefruit peel polysaccharide (I). DETAILED DESCRIPTION

[0027] The following examples are intended solely to illustrate the present invention and are not intended to limit the scope of this application, which is defined solely by the claims. This application presents only representative experimental data. Due to space limitations, some data, particularly the initial exploration of parameters such as concentration, dosage, pH, and temperature, are not fully presented.

[0028] Example 1 Materials and Basic Experimental Methods

[0029] Extraction method of grapefruit peel polysaccharide

[0030] The grapefruit peel polysaccharide used in the experiments shown in the examples was homemade, and the raw materials were prepared using the peels of the same batch of commercially available pomelo from the same brand / distributor.

[0031] Based on preliminary research, the preparation method is as follows: grapefruit peel is dried, crushed, and passed through a 100-mesh sieve; deionized water is added at a material-liquid ratio of 1:30 (mass / volume), and the pH is adjusted to 5.0; extraction is performed at 100°C for 2 hours; after cooling, the extract is filtered; precipitated by adding 3 volumes of ethanol; the precipitate is centrifuged, concentrated, and freeze-dried. The yield of grapefruit peel polysaccharide by weight is approximately 12%.

[0032] Subsequently, the applicant purchased two commercially available grapefruit peel polysaccharides (Shaanxi Hana Biological and Lanzhou Waters Biological), and treated them using the preferred method in the examples, achieving similar results.

[0033] Preparation and use of psoriasis cell models

[0034] Methods modified from "Effects of QHJD on TNF-α-induced proliferation and apoptosis of HaCat cells." Chinese Journal of Experimental Traditional Chinese Medicine, Vol. 22, No. 4, 2016. Based on the rapid proliferation of epidermal cells induced by TNF-α, similar to psoriasis:

[0035] Immortalized human epidermal cells HaCat cells were used to prepare a psoriasis cell model (HaCat cells and special culture medium were purchased from Wuhan Sean Biotechnology, and the cell culture and passaging methods were carried out according to the operating instructions provided by the company).

[0036] HaCat cells in the logarithmic growth phase were taken, rinsed with PBS, and digested with 0.25% trypsin for 5 minutes. Culture medium was added to terminate the digestion and the cells were pipetted into a cell suspension. The cells were washed by centrifugation and resuspended in culture medium. The resuspended cells were inoculated into the wells of a 96-well plate, with 200 μl per well, approximately 5,000 cells.

[0037] The wells were divided into a blank group, a control group, and a test drug group; after culturing on a 96-well plate for 24 hours, the culture medium containing TNF-α (40 ng / mL) was replaced for the HaCat cells in the control group and the test drug group, and the blank group used a culture medium without TNF-α for 24 hours; the culture medium containing the test drug was replaced with the test drug at the desired concentration, and the culture medium without TNF-α and the test drug was replaced for the blank group and the control group; after culturing for 24 hours, a CCK-8 experiment was performed to detect cell activity / survival rate (CCK-8 kit from CLPBIO, performed according to the operating instructions provided by the kit), and the cell survival rate of the blank group was calculated as 100%, and the relative survival rate of the control group and the test drug group was calculated.

[0038] Preparation and use of psoriasis mouse models

[0039] This method was modified from "Hesperidin inhibits keratinocyte proliferation and imiquimod-induced psoriasis-like dermatitis via the IRS-1 / ERK1 / 2 pathway, LifeScience, 2019, Vol. 219"; based on imiquimod / IMQ-induced psoriasis-like skin problems:

[0040] A 2cm×2cm area of ​​skin was prepared from the backs of healthy BALB / C mice, especially those without obvious skin problems after examination. The mice were divided into a blank group and a model group. Vaseline cream and 5% IMO cream were applied once a day, respectively. The skin condition was observed after 1 week. A successful model was considered if wrinkles, keratinization, redness, swelling, and white scales were observed compared with those of the blank mice.

[0041] The following criteria were used for grouping to ensure that the initial scores within each group were essentially the same. Dosing was continued for 7 consecutive days. The prepared skin was cleansed (gently massaged) with body wash for 3 minutes once daily, and the body wash was thoroughly washed off with purified water 20 minutes after washing. Scoring was performed before dosing (day 0) and 24 hours after the last dose (day 7). Scoring was based on the PASI criteria for psoriasis. Since the model area was already determined, only four criteria were evaluated: skin lesions, scaling, plaque hypertrophy, and erythema.

[0042] Table 1 Scoring criteria for mouse psoriasis model (referenced to PASI)

[0043]

[0044] DPPH scavenging rate assay and superoxide anion radical scavenging rate assay

[0045] The DPPH scavenging rate and superoxide anion radical scavenging rate of 0.5 mg / mL grapefruit peel polysaccharide solution were determined using a commercially available DPPH scavenging rate assay kit (Shanghai Yanqi Biological) and a superoxide anion radical scavenging rate assay kit (Haisai Peisen Biological).

[0046] Formula and preparation method of shower gel

[0047] Shower gel formula (by weight): 50 parts of deionized water, 10 parts of sodium laureth sulfate, 10 parts of ammonium laureth sulfate, 5 parts of ammonium lauryl sulfate, 5 parts of cocamide, 10 parts of cocamidopropyl betaine, 2 parts of glycerin, 3 parts of sodium lauroyl sarcosinate, 2 parts of acrylic copolymer, 0.2 parts of fragrance, 2 parts of citric acid, 2 parts of glossing agent, 4 parts of grapefruit peel polysaccharide.

[0048] Preparation method: Heat to 80 degrees Celsius in an emulsifying pot and slowly stir and add deionized water, acrylic copolymer, sodium laureth sulfate, ammonium laureth sulfate, citric acid, glycerin, ammonium lauryl sulfate, and cocamide in sequence; cool to 50 degrees Celsius and add cocamidopropyl betaine and sodium lauroyl sarcosinate; cool to 35 degrees Celsius and add gloss agent, flavor, and grapefruit peel polysaccharide.

[0049] Example 2 Enzyme modification experiment on grapefruit peel polysaccharide

[0050] The following enzyme modification experiments were performed using the same batch of grapefruit peel polysaccharides prepared as described in Example 1 (condition exploration experiments are not shown): The enzymes used in Example 2 were all food grade and purchased from Suzhou Zhuoxin or Beijing Xiasheng.

[0051] The polysaccharide concentration was prepared to 5 mg / mL. Based on previous experiments, pectinase, α-amylase, and β-amylase were added to 100 U / mL, pH 6.0, and treated at 40 degrees Celsius for 2 hours, or polygalacturonase, pectinesterase, and pectin lyase were added to 80 U / mL, pH 6.0, and treated at 40 degrees Celsius for 2 hours, or arabinofuranosidase and xylanase were added to 200 U / mL, pH 7.0, and treated at 40 degrees Celsius for 3 hours. After treatment, the enzymes were inactivated at 80 degrees Celsius for 3 minutes. After cooling, the mixture was re-concentrated and freeze-dried.

[0052] Example 3 Effect of enzyme modification on the antioxidant effect of grapefruit peel polysaccharide and the effect of treating psoriasis cell model

[0053] The effect of grapefruit peel polysaccharide on psoriasis cell model was determined by the method described in Example 1. Combined with the observation of culture medium viscosity in the previous experiment, the polysaccharide concentration in the culture medium was selected to be 20 mg / mL. The results are as follows: Figure 1 As shown in the results, grapefruit peel polysaccharide can effectively inhibit the growth of HaCat cells. Among the enzyme-modified polysaccharides, except for arabinofuranosidase, which is basically ineffective, the modification of several other enzymes can improve the inhibitory effect compared with the unmodified polysaccharide, especially pectinase and polygalacturonase.

[0054] The antioxidant effects of grapefruit peel polysaccharide and enzyme-modified grapefruit peel polysaccharide were determined using the method described in Example 1. Figure 2 As shown, compared to unmodified polysaccharides, modifications with other enzymes all enhanced the antioxidant activity of polysaccharides, with polygalacturonase being particularly effective (although its efficacy in cell-based experiments was inferior to that of the more widely used pectinase). Considering antioxidant activity as a key indicator for topical medications for psoriasis, we selected pectinase- and polygalacturonase-modified grapefruit peel polysaccharides and unmodified grapefruit peel polysaccharides to prepare shower gels for the following experiments.

[0055] Example 4 Therapeutic Effect of Grapefruit Peel Polysaccharide on Psoriasis Mouse Model

[0056] Shower gel products containing no grapefruit peel polysaccharide, grapefruit peel polysaccharide modified with pectinase, grapefruit peel polysaccharide modified with polygalacturonase, and unmodified grapefruit peel polysaccharide were prepared according to the description in Example 1. Furthermore, experiments on a psoriasis mouse model were conducted according to the protocol described in Example 1. The results are shown in Table 2:

[0057] Table 2 Scores of the mouse psoriasis model after 7 days of treatment (n=10 per group, all survived)

[0058]

[0059]

[0060] The results preliminarily demonstrated the feasibility of using grapefruit peel polysaccharides in a shower gel for daily psoriasis treatment. Furthermore, grapefruit peel polysaccharides modified with pectinase and polygalacturonase enzymes demonstrated superior efficacy compared to unmodified grapefruit peel polysaccharides. In particular, modification with polygalacturonase enhanced both antioxidant activity and therapeutic efficacy. Subsequent trials with psoriasis patients also confirmed the effectiveness of this product.

Claims

1. Application of enzyme-modified grapefruit peel polysaccharide in the preparation of a medicament for treating psoriasis, wherein the medicament is in the form of a shower gel. The enzyme-modified grapefruit peel polysaccharide is prepared according to the following method: (1) Prepare grapefruit peel polysaccharide at a concentration of 5 mg / mL; (2) Add polygalacturonase to 80 U / mL, pH 6.0, and treat at 40-60 degrees Celsius for 2 hours; (3) inactivating the enzyme; cooling, concentrating and freeze-drying; The grapefruit peel polysaccharide is prepared according to the following method: the grapefruit peel is dried, crushed, and passed through a 100-mesh sieve; deionized water is added according to a material-liquid ratio of 1:30 by mass / volume, and the pH is adjusted to 5.0; extraction is performed at 100 degrees Celsius for 2 hours; after cooling, the polysaccharide is filtered; precipitated by adding 3 times the volume of ethanol; the precipitate is collected by centrifugation, concentrated, and freeze-dried.

2. The use according to claim 1, wherein the shower gel further comprises an ingredient selected from surfactants, moisturizers, fragrances, and glossing agents.

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