Application of algin salicylate and application of algin salicylate in acne-removing medical apparatus or skin care product

Abstract

The invention relates to preparation and application of algin salicylate. The algin salicylate is prepared by reacting algin and salicylic acid under the catalysis of EDC and DMAP. The alginate salicylate disclosed by the invention firstly has an excellent antibacterial effect, and can inhibit the growth of propionibacterium acnes and staphylococcus aureus causing skin acnes; the traditional Chinese medicine composition has the advantages that the traditional Chinese medicine composition has good moisturizing and grease secretion inhibiting performance, can regulate water-oil balance of skin, can avoid water loss of the skin, can regulate excessive grease secretion, is low in cytotoxicity, irritation and inflammability, and can avoid irritation sensitivity caused by damage of a skin barrier. Algin salicylate is free of immunogenicity, good in air permeability, capable of keeping the moist environment of the skin for a long time, safe, non-toxic and convenient to apply. The acne-removing composition can be added into medical instruments or cosmetics with an acne-removing function, and has a good acne-removing effect.

Description

Technical Field

[0001] The invention relates to the field of preventing and treating acne skin diseases, and in particular to an application of alginate salicylate. Background Art

[0002] The skin is the largest organ in the human body. It has many functions, such as maintaining body temperature, preventing water loss, and acting as a barrier to protect the body from damaging pathogens, pollution, and ultraviolet (UV) radiation. Acne vulgaris, also known as acne vulgaris or commonly known as "acne", is a common chronic inflammatory skin disease that mainly occurs on the cheeks, forehead, and mandible, and can also extend to the trunk such as the front chest, back, and shoulder blades. The overall presentation is characterized by acne, papules, pustules, nodules, cysts, and scars, accompanied by sebum secretion. The high-risk population is mainly adolescent boys and girls, with an incidence rate of up to 54%. The causes of acne are complex, including abnormal androgen levels in the body, excessive secretion of sebaceous glands, abnormal keratinization of cells around hair follicles, and inflammatory reactions caused by Propionibacterium acnes. Acne care should first inhibit the excessive proliferation of Propionibacterium acnes, Staphylococcus aureus, and Staphylococcus epidermidis, and secondly inhibit the excessive secretion of skin oil that causes pore blockage. Attention should also be paid to skin water content and strengthening skin barrier function.

[0003] Compared with terrestrial plants, seaweed lives in intertidal areas with high osmotic and high salinity and large waves. Due to the particularity of the living environment, the cell walls of seaweed are rich in polysaccharides to resist environmental damage. Polysaccharides that protect the algae also play an important role in protecting human skin. Alginate is an acidic polysaccharide derived from brown algae, mainly composed of mannuronic acid and / or guluronic acid, and has multiple biological activities such as antioxidant, moisturizing, anti-inflammatory, and anti-aging. Alginate salicylate generated by the reaction of alginate and salicylic acid has the following repeating structural units:

[0004]

[0005] The alginate salicylate of the present invention first has an excellent antibacterial effect, and can inhibit the growth of Propionibacterium acnes and Staphylococcus aureus that cause skin acne; secondly, it has good moisturizing and oil secretion inhibition performance, can regulate the water-oil balance of the skin, can avoid skin water loss, and can regulate excessive oil secretion. In addition, it has low cytotoxicity, irritation and inflammatory properties, and can avoid irritation and sensitivity caused by skin barrier damage. Algin salicylate has no immunogenicity, good air permeability, can maintain the moist environment of the skin for a long time, is safe and non-toxic, and is easy to use.

[0006] The invention can be added into medical devices or cosmetics with acne-removing function and has good acne-removing effect. Summary of the invention

[0007] The present invention aims at the deficiencies of the prior art and provides an application of alginate salicylate in acne care, characterized in that the application refers to the application of alginate salicylate as an active ingredient in the preparation of medical devices or skin care products with acne removal function.

[0008] To achieve the above object, the technical solution adopted by the present invention is: an application of alginate salicylate, characterized in that the application refers to the application of alginate salicylate as an active ingredient in the preparation of medical devices or skin care products with acne removal function. The structure of the alginate salicylate is as follows:

[0009]

[0010] Among them, the medical device or skin care product with acne removal function is characterized in that alginate salicylate is used as the active ingredient.

[0011] Wherein, the alginate salicylate is characterized in that the alginate salicylate is prepared by the following method: 2-5 g of sodium alginate is added to a mixed solution of 30-80 mL of 0.6-0.8 M hydrochloric acid and 30-80 mL of 95% ethanol, and stirred continuously. Stand at 4°C overnight. After vacuum filtration, wash the solid with 95% ethanol 2-3 times, put it in an oven at 50-60°C to dry to obtain solid alginate. Disperse the dried alginate in 100-150 mL of water, and slowly add 5-12 mL of 40% tetrabutylammonium hydroxide until the alginate is completely dissolved. Adjust the pH to 7.0-8.0. The solution is dialyzed overnight in a 12000 Da dialysis bag, concentrated by rotary evaporation, and freeze-dried to obtain solid tetrabutylammonium alginate. 1-3 g of salicylic acid is dissolved in 100-300 mL of N,N-dimethylformamide (DMF), stirred until completely dissolved, 1-3 g of 1-ethyl-(3-dimethylaminopropyl)-carbodiimide (EDC) and 0.33-1 g of 4-dimethylaminopyridine (DMAP) are mixed and added and stirred for 15-30 minutes to activate the carboxyl group of salicylic acid. 1-3 g of tetrabutylammonium alginate is dissolved in DMF, and 5% tetrabutylammonium fluoride (TBAF) is added to the above mixed solution. The reaction system is protected by N2 and reacted at 45-50°C overnight. After the reaction is completed, water is added to the reaction system until the polysaccharide ester is precipitated, the upper layer product is collected, washed with water, centrifuged, and freeze-dried. The obtained product is ground into powder to obtain alginate salicylate SA-g-ALG.

[0012] The alginate salicylate is characterized in that the alginate salicylate is alginate salicylate of 1-4 linked mannuronic acid and / or guluronic acid with a degree of polymerization of ≥100, preferably 200-250, and the degree of substitution of salicylic acid is 15-35%.

[0013] The alginate is characterized in that the alginate is homopolymerized guluronic acid with a degree of polymerization of ≥100 or homopolymerized mannuronic acid with a degree of polymerization of ≥100, or a hybrid alginate containing both monosaccharides with a degree of polymerization of ≥50.

[0014] Among them, the medical device or skin care product for removing acne is characterized in that the medical device or skin care product refers to an external preparation for the skin surface with acne removal effect, such as cream, lotion, gel, spray, etc.

[0015] In the medical device or skin care product for removing acne, alginate salicylate is added as an active ingredient into a matrix such as water or glycerin to prepare a medical device or skin care product with acne removing function, and the added amount is 0.5-3%.

[0016] The present invention finds that the alginate salicylate has antibacterial, anti-inflammatory, moisture absorption and moisturizing, and oil secretion regulating activities, and can be used for acne removal. The application refers to the application of alginate salicylate as an active ingredient in the preparation of medical devices or skin care products with acne removal function.

[0017] The alginate salicylate of the present invention first has an excellent antibacterial effect, and can inhibit the growth of Propionibacterium acnes and Staphylococcus aureus that cause skin acne; secondly, it has good moisturizing and oil secretion inhibition performance, can regulate the water-oil balance of the skin, can avoid skin moisture loss, and can regulate excessive oil secretion. In addition, secondly, the cytotoxicity, irritation and inflammatory properties are low, and the irritation sensitivity caused by skin barrier damage can be avoided. Algin salicylate has no immunogenicity, good air permeability, and can maintain the moist environment of the skin for a long time, is safe and non-toxic, and is easy to use. The present invention is particularly suitable for removing acne.

[0018] Compared with the prior art, the present invention has the following advantages:

[0019] 1. The alginate salicylate prepared by the present invention combines the skin activity of alginate and salicylic acid, has excellent antibacterial, anti-inflammatory, moisture absorption and moisturizing and oil secretion regulating activities, can maintain the moist environment of the skin for a long time, and exert a long-lasting effect;

[0020] 2. It is hydrophilic and lipophilic, and can better penetrate and stay in the hair follicles and sebaceous glands of the skin, effectively reducing skin inflammation and better repairing the skin barrier.

[0021] 3. The method of the present invention is simple, the experimental materials are easily available, and it is convenient for mass preparation. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 UV spectrum of alginate salicylate

[0023] Figure 2IR spectrum of alginate salicylate.

[0024] Figure 3 Alginate Salicylate 1 H-NMR spectrum.

[0025] Figure 4 Moisturizing properties of alginate salicylate.

[0026] Figure 5 Antibacterial activity of alginate salicylate. (a) is the MIC of Propionibacterium acnes, (b) is the antibacterial curve of Propionibacterium acnes, (c) is the MIC of Staphylococcus aureus, and (d) is the antibacterial curve of Staphylococcus aureus.

[0027] Figure 6 Inhibitory effect of alginate salicylate on 5α-reductase.

[0028] Figure 7 Alginate salicylate is toxic to HaCaT cells.

[0029] Figure 8 Hemolytic effect of alginate salicylate on blood cells.

[0030] Fig. 9 Anti-inflammatory effects of alginate salicylate on zebrafish with LPS-induced inflammation. DETAILED DESCRIPTION

[0031] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention. In addition, it should be understood that after reading the content of the present invention, those skilled in the art may make various changes or modifications to the present invention, and these equivalent forms fall within the scope limited by the present invention equally.

[0032] Example 1, Preparation of Salicylic Grafted Alginate (SA-g-ALG)

[0033] (1) Preparation of tetrabutylammonium alginate (TBA-ALG, TBA-alginate)

[0034] 2g sodium alginate (Mw = 1000kDa, M / G = 3.73) was added to a mixed solution of 30mL 0.6M hydrochloric acid and 30mL 95% ethanol by volume, and stirred continuously. Let stand overnight at 4°C. After vacuum filtration, the solid was washed 3 times with 95% ethanol by volume, and dried in an oven at 50°C to obtain solid alginate. The dried alginate was dispersed in 100mL water, and 5mL of 40% tetrabutylammonium hydroxide aqueous solution was slowly added until the alginate was completely dissolved. The pH was adjusted to 7.0-8.0 with 20g / L dilute hydrochloric acid. The solution was placed in a 12000Da dialysis bag and dialyzed in water overnight for 20 hours. After rotary evaporation and freeze-drying, solid tetrabutylammonium alginate was obtained.

[0035] (2) Preparation of algin salicylate (SA-g-ALG)

[0036] 1g salicylic acid was dissolved in 100mL N,N-dimethylformamide (DMF), stirred until completely dissolved, 1g 1-ethyl-(3-dimethylaminopropyl)-carbodiimide (EDC) and 0.33g 4-dimethylaminopyridine (DMAP) were mixed and added and stirred for 20min to activate the carboxyl group of salicylic acid to obtain salicylic acid solution. 1, 2, and 3g tetrabutylammonium alginate were dissolved in 100mL DMF, and 1, 2, and 3mL of 5% tetrabutylammonium fluoride (TBAF) were added to the above salicylic acid solution. The reaction system was protected by N2 and reacted at 45-50℃ overnight. After the reaction was completed, water was added to the reaction system until the polysaccharide ester precipitated, the upper layer product was collected, washed with water, centrifuged, and freeze-dried. Alginate salicylate was obtained, and the obtained product was ground into powder to obtain algin salicylate, which were successively recorded as SA-g-ALG-1, SA-g-ALG-2 and SA-g-ALG-3. The yields were 28%, 33% and 40% respectively.

[0037] Example 2, Chemical Composition Analysis and Structural Characteristics of Algin Salicylate

[0038] Salicylic acid was prepared into 0.06, 0.07, 0.08, 0.09, 0.1 mg / mL standard solutions. After adding 0.5 mol / LFeCl3 solution, the phenolic hydroxyl group of salicylic acid was ionized to produce C6H5O - and Fe 3+The formed complex H3[Fe(OC6H5)6] is purple, and the absorbance at 520nm is measured, and a calibration curve is made (the horizontal axis is the salicylic acid concentration, and the vertical axis is the absorbance). SA-g-ALG-1, SA-g-ALG-2 and SA-g-ALG-3 prepared in Example 1 are respectively prepared with 0.5 mg / mL alginate salicylate aqueous solution, and the absorbance is measured in the same way (substituting into the above calibration curve to obtain the salicylic acid group concentration) and the grafting rate is calculated. The grafting rate represents the grafting density, that is, the weight of grafted salicylic acid groups in a unit weight of alginate salicylate. The grafting rate of alginate salicylate prepared in Example 1 is shown in Table 1.

[0039] Table 1 Grafting rate of alginate salicylate

[0040]

[0041] The grafting rates of SA-g-ALG-1, SA-g-ALG-2 and SA-g-ALG-3 increase in sequence. A high grafting rate means that the weight of grafted salicylic acid molecules per unit weight of polymer is greater, and its biological activity is higher. Therefore, the highly substituted alginate salicylate SA-g-ALG-3 prepared in Example 1 was characterized and its anti-acne activity was determined.

[0042] SA-g-ALG-3 was characterized by UV, IR and one-dimensional hydrogen spectra. Figure 1 , IR spectrum see Figure 2 , 1 H-NMR images are shown in Figure 3 .

[0043] The UV spectrum shows that compared with the raw material TBA-ALG, SA-g-ALG-3 has an absorption peak in the UV region, indicating the presence of a conjugated structure of the benzene ring; the infrared spectrum shows an increase in the ester group at 1745 cm -1 、1726cm -1 The absorption peak is 1745cm -1 、1726cm -1 It is the stretching vibration of the ester group C=O, indicating that the esterification is successful. 1 From the H NMR spectrum, we can see that alginate salicylate has more small peaks between 6.0-8.0 ppm than the raw material, and 6.0-8.0 ppm is the chemical shift of benzene ring hydrogen, which also shows that alginate salicylate is successfully prepared.

[0044] Example 3, Moisturizing properties of algin salicylate SA-g-ALG-3

[0045] Test samples: alginate salicylate SA-g-ALG-3 prepared in Example 1, salicylic acid.

[0046] Test method:

[0047] Moisture retention: The determination of moisture retention needs to be carried out under a constant temperature and humidity environment. Dry the volumetric flask at 115°C to constant weight in advance, and place the sample in a drying oven after freeze-drying for later use. Weigh 0.3g of dried SA-g-ALG-3 and salicylic acid respectively and place them in a dry (height × inner diameter) 50×30mm weighing bottle, add three times the mass of distilled water to mix thoroughly, weigh it as M1, place it in a desiccator, maintain the ambient temperature at 29°C, RH = 19%, weigh it at 0, 4, 8, 12, 24, 36, 48, 60, 72h and record it as Mt, and determine the amount of water reduction. Hyaluronic acid was selected as the positive control, and the formula for calculating the moisture retention rate is:

[0048] Moisture retention (g / g) = sample weight at time t Mt / initial sample weight M1.

[0049] result:

[0050] Alginate Salicylate Moisturizing Figure 4 .

[0051] The water retention of alginate salicylate under low humidity (19%) conditions is significantly better than that of salicylic acid. The moisturizing rates of salicylic acid and alginate salicylate at 4, 8, 12, 24, 36, 48, 60 and 72 hours are maintained at 86.41%, 89.86%, 74.90%, 79.42%, 63.92%, 70.35, 0, 18.15%, 0, 3.89%, 0, 0.99%, 0, 0.57%, respectively. The moisturizing rate of alginate salicylic acid is continuously better than that of salicylic acid during 72 hours. And after 24 hours, the moisturizing rate of salicylic acid is 0, while alginate salicylic acid still has moisturizing properties. This shows that alginate salicylate has good moisturizing properties, and its moisturizing time and moisturizing rate are better than those of salicylic acid.

[0052] Example 4, Antibacterial Activity of Algin Salicylate SA-g-ALG-3

[0053] Test sample: Algin salicylate SA-g-ALG-3 prepared in Example 1, solutions of different concentrations were prepared.

[0054] Test method:

[0055] The MIC of Propionibacterium acnes was determined by microplate dilution method. 6100 μL of CFU / mL Propionibacterium acnes suspension was added to a 96-well plate, and the final concentration of SA-g-ALG-3 was adjusted to 5, 4, 3, 2, and 1 mg / mL. Salicylic acid (instead of sample solution) with a final concentration of 2 mg / mL was used as a positive control, and sterile distilled water (instead of sample solution) was used as a negative control (representing the growth of bacteria without sample (salicylic acid or alginate salicylate) intervention). Each group was repeated 3 times. After 24 hours of culture, the lowest concentration at which the bacterial suspension did not become turbid was observed, which was the MIC.

[0056] The MIC of Staphylococcus aureus was measured by resazurin combined with microplate dilution method. 6 100L of CFU / mL Staphylococcus aureus suspension was added to a 96-well plate, and the final concentration of SA-g-ALG-3 was adjusted to 5, 4, 3, 2, and 1mg / mL. Salicylic acid (instead of the sample solution) with a final concentration of 2mg / mL was used as the positive control, and sterile distilled water (instead of the sample solution) was used as the negative control (representing the growth of bacteria without sample (salicylic acid or alginate salicylate) intervention). There were 3 replicates in each group. After culturing for 16 to 18 hours (18 hours in this case), 20L of 0.2% resazurin solution was added. Observe the color change of the bacterial suspension. The lowest concentration that does not change to pink is the minimum inhibitory concentration. Take the bacterial suspension in the logarithmic growth period and dilute the OD with culture medium. 600 =0.1, add samples to the final concentration of MIC, 1 / 2MIC and 1 / 4MIC, salicylic acid with a final concentration of 2mg / mL as the positive control, and distilled water as the negative control. After adding the samples, the bacterial solution was placed at 37°C for static culture. Propionibacterium acnes was sampled at 0, 24, 36, 48, 60, and 72h, and Staphylococcus aureus was sampled at 0, 2, 4, 6, 8, and 10h of culture. The absorbance at 600nm was measured by an ELISA instrument. With time as the horizontal axis, OD 600 The vertical axis is used to draw the inhibition growth curve.

[0057] result:

[0058] The minimum inhibitory concentration (MIC) and inhibition rate of alginate salicylate SA-g-ALG-3 against Staphylococcus aureus and Propionibacterium acnes are shown in Figure 5 .

[0059] The minimum inhibitory concentration MIC of alginate salicylate SA-g-ALG-3 against Staphylococcus aureus is 5 mg / mL, and the 10h inhibition rates of MIC, 1 / 2MIC and 1 / 4MIC are 100%, 71% and 51% respectively. The minimum inhibitory concentration MIC of alginate salicylate SA-g-ALG-3 against Propionibacterium acnes is 4 mg / mL, and the 24h inhibition rates of MIC, 1 / 2MIC and 1 / 4MIC are 98%, 89% and 87% respectively.

[0060] Example 5, Inhibitory effect of algin salicylate SA-g-ALG-3 on 5α-reductase

[0061] Test sample: Algin salicylate SA-g-ALG-3 prepared in Example 1.

[0062] Experimental methods:

[0063] The activity of 5α-reductase after the reaction with alginate salicylate SA-g-ALG-3 was determined by human (Human) 5α-reductase (5AR) ELISA kit (purchased from Shanghai ELISA Biotechnology Co., Ltd.) to determine its lipid metabolism intervention activity. SA-g-ALG-3 was dissolved in PBS (buffer) to prepare an 8 mg / mL solution. Standard wells, sample wells and blank wells were set up, and 50 μL of 5α-reductase with concentrations of 0, 12.5, 25, 50, 100, and 200 pg / mL was added to each standard well, 25 μL of 100 pg / mL of 5α-reductase was added to the sample well, and 25 L of alginate salicylate SA-g-ALG-3 and 2 mg / mL of salicylic acid were added to the blank well, and 50 μL of PBS buffer was added to the blank well. The strips were equilibrated at room temperature for 20 minutes before adding the sample. Except for the blank wells, add 100 μL of horseradish peroxidase (HRP)-labeled detection antibody to each well of the standard well and sample well, seal the reaction well with a sealing film, and incubate in a constant temperature oven for 60 minutes. Discard the liquid, pat dry on absorbent paper, fill each well with washing solution (provided by the kit), let stand for 1 minute, shake off the washing solution, pat dry on absorbent paper, and repeat the washing plate 5 times. Add 100 μL of substrate 3,3',5,5'-tetramethylbenzidine (TMB) to each well and incubate at 37°C in the dark for 15 minutes. Add 50 μL of stop solution to each well, and measure the OD value of each well at a wavelength of 450 nm within 15 minutes. Make a standard curve and calculate the inhibition rate. The inhibition rate calculation formula is:

[0064] Inhibition rate (%) = (OD standard well - OD sample well) / (OD standard well - OD blank well) × 100%.

[0065] result:

[0066] The inhibitory effect of SA-g-ALG-3 on 5α-reductase Figure 6 .

[0067] The inhibition rates of SA-g-ALG at 4, 2, and 1 mg / mL alginate salicylate SA-g-ALG-3 were 46.13%, 37.02%, and 30.56%, respectively, indicating that it has significant activity in inhibiting 5α-reductase and can inhibit the secretion of oil.

[0068] Example 6, HaCaT cell toxicity test of alginate salicylate SA-g-ALG-3

[0069] Test sample: Algin salicylate SA-g-ALG-3 in Example 1.

[0070] Test method:

[0071] Dilute the sample to 800, 600, 400 and 200 μg / mL with complete culture medium, for a total of 4 gradient concentrations. Take a 25 mL cell culture flask cultured to the logarithmic phase of HaCaT cells, wash twice with PBS, add 1 mL of trypsin for digestion, centrifuge at 11000 rpm for 4 min, discard the supernatant, add 1 mL of complete culture medium (DMEM high glucose medium) containing 10% fetal bovine serum, and pipette evenly. Take 10 μL and count with a hemocytometer, according to 1*10 4 Cells / well (100 μL per well) were inoculated in a 96-well plate. After incubation for 24 hours, the culture medium was discarded, and 100 μL of samples of different concentrations (800, 600, 400 and 200 μg / mL) diluted with the above culture medium were added to each well of the treatment group. Only 100 μL of complete culture medium was added to the blank group without any sample. Each experimental group was repeated for 3 wells and incubated for another 24 hours. After adding 10 μL of MTT solution (5 mg / mL) to each well, incubate at 37°C for another 4 hours. Observe the appearance of purple crystals at the bottom of the 96-well plate, remove the 96-well plate, gently aspirate and discard the supernatant, add 150 μL of DMSO to each well and shake on a shaker for 10 minutes until the crystals dissolve. The mixed solution was measured at 570 nm using an enzyme reader. The cell viability of the blank group was set as 100%, and the cell viability of the treatment group was calculated relative to the blank group.

[0072] Cell viability (%) = absorbance of cells in the treated group / absorbance of cells in the blank group*100%.

[0073] Results: MTT test results are shown in Figure 7 .

[0074] After 800, 600, 400 and 200 μg / mL of salicylic acid and alginate salicylate were applied to HaCaT cells, the cell survival rates were 48.84%, 68.57%, 71.82%, 90.74%, 83.47%, 98.33%, 123.11% and 114.60%, respectively. The cell survival rate of alginate salicylate was higher than that of salicylic acid. 800 μg / mL of salicylic acid and alginate salicylate were cytotoxic, but alginate salicylate mitigated the cytotoxicity. The cell survival rates of 600, 400 and 200 μg / mL of salicylic acid were all greater than 70%, with no significant cytotoxicity.

[0075] Example 7, Hemolytic Activity of Algin Salicylate SA-g-ALG-3 on Blood Cells

[0076] Test sample: Algin salicylate SA-g-ALG-3 in Example 1.

[0077] Test method:

[0078] 10 mL of blood was collected from the ear vein of New Zealand white rabbits and placed in an anticoagulant centrifuge tube. The blood was centrifuged at 2000-2500 rpm for 15 min, and washed with sterile saline for 3 times until there was no transparent supernatant to obtain blood cells. The red blood cells were prepared into a 4% red blood cell suspension with sterile saline. Algin salicylate SA-g-ALG-3 was prepared into a μg / mL solution with sterile saline. 100 μL of sample (salicylic acid and alginate SA-g-ALG-3) was added to 100 μL of 4% red blood cell suspension to a final concentration of 1200, 300, 150, and 75 μg / mL. 100 μL of sterile saline (instead of sample) was used as a negative control, and 100 μL of distilled water (instead of sample) was used as a positive control. After standing at 37°C for 3 hours, the supernatant was centrifuged at 1000 rpm. The absorbance was measured at 540 nm by an enzyme marker.

[0079] Hemolysis rate (%) = (OD sample - OD negative) / (OD positive - OD negative) × 100%

[0080] Results: Hemolysis test results are shown in Figure 8 .

[0081] The experimental results show that the red blood cell hemolysis rates of salicylic acid and alginate salicylate at 1200, 300, 150, and 75 μg / mL are 14.53%, 4.90%, 7.07%, 4.81%, 6.60%, 4.71%, 5.56%, and 3.92%, respectively. At 1200 μg / mL, the hemolysis rate of alginate salicylate is significantly reduced, and the hemolysis rates of alginate salicylate at each concentration are all lower than 5%, indicating that alginate salicylate has good biocompatibility.

[0082] Example 8, Anti-inflammatory Activity of Algin Salicylate SA-g-ALG-3

[0083] Test sample: Algin salicylate SA-g-ALG-3 in Example 1.

[0084] Experimental method: Zebrafish were raised in fish water at 28°C (water quality: 200mgNaCl was added to every 1L of reverse osmosis water). 2dpf transgenic neutrophil fluorescent zebrafish (MPX) (purchased from the National Zebrafish Resource Center of the National Aquatic Biological Germplasm Resource Bank) were randomly selected in a 6-well plate, and 30 zebrafish were treated in each well (experimental group). 15 fish per well. The water-soluble sample was given, the final concentration of salicylic acid was 50ug / mL, and the final concentration of alginate salicylate SA-g-ALG-3 was 25, 50, 100μg / mL. At the same time, a normal control group (no modeling, that is, no injection of lipopolysaccharide to establish a zebrafish inflammation model) and a model control group (modeling without drug administration, that is, injection of lipopolysaccharide-based zebrafish inflammation model, but no sample treatment) were set up, each well had a capacity of 3mL, and incubated in the dark at 28°C for 1h. After the incubation, lipopolysaccharide (LPS) was injected to establish the zebrafish inflammation model, and the cells were incubated in the dark for 2 hours at 28°C. Ten zebrafish were randomly selected from each experimental group and photographed under a fluorescence microscope. Advanced image processing software was used to analyze and collect data, and the number of zebrafish yolk sac neutrophils (N) was analyzed. The formula was used to calculate and determine whether the sample had anti-inflammatory effects.

[0085] Anti-inflammatory efficacy (%) = (N1-Ns) / (N1-N0) × 100%. N1 is the model control group, N0 is the normal control group, and Ns is the sample group.

[0086] Results: The experimental results of zebrafish inflammation model are shown in Fig. 9 .

[0087] Algin salicylate SA-g-ALG-3 can significantly inhibit inflammation in zebrafish at concentrations of 0.0025%, 0.005 and 0.01% (i.e. 25, 50, 100 μg / mL), with inhibition rates of 22%, 25% and 39% respectively, showing significant anti-inflammatory effects.

[0088] The above is a description of the embodiments of the present invention. Through the above description of the disclosed embodiments, professionals and technicians in the field can implement or use the present invention. Various modifications to these embodiments will be obvious to professionals and technicians in the field. The general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but will conform to the widest range consistent with the principles and novel features disclosed herein.

Claims

1. An application of alginate salicylate, characterized in that: The alginate salicylate can be used as an active ingredient in the preparation of medical devices or skin care products for removing acne.

2. A medical device or skin care product for removing acne, characterized in that: The acne-removing medical device or skin care product uses alginate salicylate as an active ingredient.

3. The acne-removing medical device or skin care product according to claim 2 or the use according to claim 1, characterized in that: The acne-removing medical device or skin care product uses alginate salicylate as an active ingredient; Algin salicylate is added as an active ingredient to one or more matrices such as water or glycerin to prepare a medical device or skin care product with acne removal function. The mass addition amount of algin salicylate in the medical device or skin care product with acne removal function is 0.5-3%, preferably 1.5-2.0%.

4. The acne-removing medical device or skin care product according to claim 2 or 3 or the use according to claim 1 or 3, characterized in that: The medical device refers to an external preparation for skin surface with acne removal function, such as one or more of creams, lotions, gels or sprays.

5. The use of the acne-removing medical device or skin care product or the alginate salicylate according to any one of claims 1 to 4, characterized in that: The structure of alginate salicylate is that the hydroxyl group (-OH) at the C-2 and / or C-3 position of alginate mannuronic acid or / and guluronic acid reacts with the carboxyl group (-COOH) of salicylic acid to form an ester bond, i.e., alginate salicylate substituted at the C-2 and / or C-3 position is generated; wherein the polymerization degree of alginate is ≥100, preferably 200-250; the grafting rate is 15% to 35%, and the grafting rate represents the grafting density, i.e., the weight of grafted salicylic acid groups in a unit weight of alginate salicylate, preferably 25-30%.

6. The use of the acne-removing medical device or skin care product or the alginate salicylate according to any one of claims 1 to 5, characterized in that: Alginate salicylate is prepared by the following method: 2-5 g of sodium alginate is added to a mixed solution of 30-80 mL of 0.6-0.8 M hydrochloric acid and 30-80 mL of 90%-95% ethanol by volume, and stirred continuously; 0-4℃ for 8-24 hours; After solid-liquid separation, the solid is washed with ethanol at a volume concentration of 90%-95%, and dried at 50-60°C to obtain solid alginate; The solid alginate obtained by drying is dispersed in 100-150 mL of water, and 5-12 mL of a 30%-40% tetrabutylammonium hydroxide aqueous solution is added dropwise until the alginate is completely dissolved; the pH is adjusted to 7.0-8.0 with 10-20 g / L dilute hydrochloric acid; the solution is placed in a 10000-12000 Da dialysis bag and dialyzed in water for 8-24 hours, and the material in the dialysis bag is dried to obtain solid tetrabutylammonium alginate; 1-3g salicylic acid is dissolved in 100-300mL N,N-dimethyl-formamide (DMF), stirred until completely dissolved, 1-3g 1-ethyl-(3-dimethylaminopropyl)-carbodiimide (EDC) and 0.33-1g 4-dimethylaminopyridine (DMAP) are added and stirred for 15-30min to activate the carboxyl group of salicylic acid to obtain salicylic acid solution; 1-3g tetrabutylammonium alginate is dissolved in 100-300mL DMF, and 1-3mL of 5-10% tetrabutylammonium fluoride (TBAF) is added to assist dissolution, and then added to the above salicylic acid solution, the reaction system is protected by N2, and reacted at 45-50°C for 8-24 hours; after the reaction is completed, water is added to the reaction system until the polysaccharide ester is precipitated, the upper layer product is collected, washed with water, and dried. Algin salicylic acid ester SA-g-ALG is obtained.

7. The use of the acne-removing medical device or skin care product or the alginate salicylate according to claim 6, characterized in that: The alginate salicylate is alginate salicylate of 1-4 linked mannuronic acid and / or guluronic acid with a polymerization degree of ≥100 (preferably 200-250), and the grafting rate of salicylic acid is 15-35%; the polymerization degree of alginate is ≥100 (preferably 200-250), and its monosaccharide composition is one or both of guluronic acid and mannuronic acid.

8. The use of the acne-removing medical device or skin care product or the alginate salicylate according to claim 6 or 7, characterized in that: The reaction formula is as follows:

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