Preparation method and permeation promotion application of high-purity sponge bone needle

Abstract

The invention discloses a preparation method and permeation promotion application of a high-purity sponge bone needle, and belongs to the technical field of sponge bone needles. Comprising the following steps: cutting a natural sponge into pieces, soaking in deionized water, carrying out ultrasonic oscillation, filtering, and cleaning to obtain a pretreated sponge; carrying out enzymolysis on the pretreated sponge with alkaline protease to obtain a primary zymolyte; and carrying out enzymolysis with ginger protease to obtain a secondary enzymolysis product, and carrying out vacuum freeze drying to obtain the high-purity sponge spicule. The natural sponge is subjected to enzymolysis treatment through alkaline protease and ginger protease, and the high-purity sponge spicule is obtained; wheat oligopeptide is grafted on the surface of a sponge spicule, and giant salamander skin collagen, a pomegranate seed extract, a lonicerae flos extract, a centella asiatica extract, an early celery extract, an iris tectorum extract, an oyster shell extract and calcium chloride mixed treatment materials are loaded, so that the repairing effect on skin inflammation acne is improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of sponge spicules, and particularly relates to a preparation method of high-purity sponge spicules and application of the high-purity sponge spicules in penetration promotion. BACKGROUND

[0002] In the field of skin care and beauty, transdermal absorption is a crucial concept, which refers to the process that active ingredients in daily cosmetics act on the surface layer or deeper tissue of the skin through a specific route. Unlike drug transdermal absorption, the transdermal absorption of functional ingredients in cosmetics focuses on effective accumulation and sustained action on the target skin layer, rather than systemic absorption. This process needs to rely on professional penetration enhancement technology, and the current mainstream penetration promotion methods mainly include physical penetration promotion and chemical penetration promotion. Among them, microneedle technology, as a representative method of physical penetration promotion, has been highly praised by the industry in recent years due to its remarkable effect. This technology forms a large number of micron-sized channels in the stratum corneum through a minimally invasive method, providing a direct delivery path for active ingredients and drug carriers to the deep layer of the skin.

[0003] Among all the penetration promotion technologies, microneedle technology is the most direct and safest. However, after the traditional solid microneedle opens the skin channel, the opened microchannels will close in a short time due to the elasticity of the skin, and the drug penetration time is limited. Although the new type of microneedle patch can be retained on the skin for a long time and continuously administer drugs, it is limited by the administration area and site due to its array type manufacturing process. The minimally invasive nature of sponge spicules makes it have little damage to the skin, almost no risk of bleeding or infection, which makes it a safe and efficient new choice for transdermal drug delivery.

[0004] Sponge spicules are usually extracted from natural sponges, but in addition to the spicule components we need, there are also a large amount of other impurities such as various organic matter, microorganisms and some minerals in natural sponges. It is not easy to completely remove these impurities during the extraction process.

[0005] Acne is a common skin disease that seriously affects the appearance and mental health of patients. The mainstream acne treatment skin care products on the market are mostly aimed at removing acne by adding components with oil control or bacteriostatic effects in skin care products. However, these components often have obvious drawbacks. The skin has limited absorption capacity, and most oil control or bacteriostatic ingredients cannot effectively penetrate, remaining on the skin surface and causing greater irritation to the skin. Long-term use can easily destroy the normal balance of skin surface flora, resulting in reduced treatment effect.

[0006] Therefore, it is urgent to develop a new type of microneedle penetration promotion technology to repair skin inflammation and acne problems. SUMMARY

[0007] The application aims to provide a preparation method of high-purity sponge bone needles and application thereof in penetration promotion, and provides a novel microneedle penetration promotion technology to repair skin inflammation and acne problems.

[0008] The application can be achieved by the following technical solutions.

[0009] A preparation method of high-purity sponge bone needles comprises the following steps.

[0010] S1, cut natural sponges, soak them in deionized water, and ultrasonically oscillate them at a frequency of 35-45 Hz for 1-2 h, filter, and wash the sponge fragments with deionized water for 4-5 times to obtain pretreated sponges;

[0011] S2, place the pretreated sponges in an extraction tank, add deionized water and alkaline protease, add a sodium hydroxide-sodium hexametaphosphate solution to adjust the pH to 10, and enzymatically hydrolyze at 40℃ for 4-6 h, then inactivate the enzyme at 110℃ for 10 min, filter, wash the solid phase to neutral, and dry to obtain a first enzymatic hydrolysate; the sodium hydroxide-sodium hexametaphosphate solution is prepared by uniformly mixing 4-8 g of sodium hydroxide, 1.8-2.2 g of sodium hexametaphosphate, and 100 mL of deionized water; the sodium hydroxide-sodium hexametaphosphate solution is used to adjust the pH suitable for alkaline protease, and at the same time, the tips of the sponge bone needles are dissolved by the alkali solution to form openings, and then hollow channels are formed, and at the same time, under the synergistic action of sodium hexametaphosphate, the OH - concentration is effectively increased, the double-layer potential is increased, and metal ion impurities in the sponge bone needles are removed;

[0012] S3, place the first enzymatic hydrolysate in an extraction tank, add deionized water and ginger protease, add a 1 mol / L hydrochloric acid solution to adjust the pH to 4.5, and enzymatically hydrolyze at 40℃ for 2-3 h, then inactivate the enzyme at 110℃ for 10 min, filter, and wash the solid phase to neutral to obtain a second enzymatic hydrolysate, and vacuum freeze-dry at -80℃ for 6-10 h to obtain a loose powder, which is the high-purity sponge bone needles.

[0013] As a further scheme of the application, in step S2, the mass ratio of the pretreated sponges, deionized water, and alkaline protease is 1-1.5:100:0.02-0.04.

[0014] As a further scheme of the application, in step S3, the mass ratio of the first enzymatic hydrolysate, deionized water, and ginger protease is 1-1.2:100:0.0075-0.014.

[0015] The high-purity sponge bone needles are prepared by the above preparation method.

[0016] The penetration-promoting application of the high-purity sponge spicules is specifically to prepare a penetration-promoting composition using the sponge spicules for repairing skin inflammation acne. The preparation method of the penetration-promoting composition comprises the following steps:

[0017] A1. Mix giant salamander skin collagen, pomegranate seed extract, honeysuckle extract, Centella asiatica extract, celery extract, iris extract, oyster shell extract, anhydrous ethanol, deionized water, and calcium chloride, and stir ultrasonically for 15-25 minutes to obtain material A;

[0018] A2, taking the high-purity sponge spicule, N,N-dimethylformamide, and deionized water, mixing, ultrasonically dispersing for 30-40 minutes, adding succinic anhydride and 3-aminopropyltriethoxysilane, heating in a water bath at 50-60°C with stirring for 5-7 hours, centrifuging, washing the solid phase, vacuum freeze-drying, and performing surface carboxylation treatment to obtain material B;

[0019] A3, taking the material B, wheat oligopeptide and deionized water, mixing them, ultrasonically treating them for 20-30 minutes, centrifuging them, washing the solid phase, drying it, and subjecting the wheat oligopeptide to an amidation reaction with the carboxyl groups on the surface of the sponge spicules to obtain material C;

[0020] A4. Mix the material C and the material A, perform ultrasonic dispersion for 30-40 min, maintain the pressure under 0.1 MPa vacuum for 30-40 min, filter, take the solid phase, and introduce the therapeutic agent containing multiple skin inflammation and acne into the hollow cavity of the sponge spicule by vacuum negative pressure treatment to obtain material D;

[0021] A5. Mix aluminum glycolate, tartaric acid, and deionized water, stir for 20-30 minutes, add sodium polyacrylate, continue stirring for 30-40 minutes, add material D, and continue stirring for 5-10 minutes to obtain the penetration-enhancing composition.

[0022] As a further scheme of the present invention, the preparation method of giant salamander skin collagen comprises the following steps: removing fat and pigment from the giant salamander skin, then taking 1g of giant salamander skin, adding 15mL of 0.5mol / L acetic acid, and then adding 2.5mg of pepsin to extract, filter and centrifuge, taking the supernatant, adding 5mol / L NaCl solution to the supernatant to make the NaCl concentration 0.9mol / L, standing for 24h to separate the layers, centrifuging to recover the flocs, placing them in ultrapure water for dialyzing for three days, changing the water every 6h, dialyzing out the acetic acid, and freeze-drying to obtain giant salamander skin collagen.

[0023] As a further scheme of the present application, in step A1, the mass ratio of the giant salamander skin collagen, pomegranate seed extract, honeysuckle extract, centella asiatica extract, early celery extract, iris extract, oyster shell extract, anhydrous ethanol, deionized water is 25-35:8-10:4-6:18-26:4-6:2-4:3-5:40-50:60-80:0.4-0.8.

[0024] As a further scheme of the present application, in step A2, the ratio of high-purity sponge bone needles, N,N-dimethylformamide, deionized water, succinic anhydride and 3-aminopropyl triethoxysilane is 0.6-0.8g:2-3mL:30mL:4-5g:8-10g.

[0025] As a further scheme of the present application, in step A3, the ratio of material B, wheat oligopeptide and deionized water is 0.4-0.8g:10-15mg:10-15mL.

[0026] As a further scheme of the present application, in step A4, the mass ratio of material C and material A is 1:10.

[0027] As a further scheme of the present application, in step A5, the ratio of ganoxylate, tartaric acid, deionized water, sodium polyacrylate and material D is 0.3-0.5g:0.4-0.6g:50mL:8-10g:1.5-2.2g.

[0028] The present application has the following advantages:

[0029] The present application uses alkaline protease and ginger protease to enzymatically hydrolyze natural sponges to obtain high-purity sponge bone needles;

[0030] Further, in terms of penetration, the hollow cavity structure of the sponge bone needle is used to press the giant salamander skin collagen, pomegranate seed extract, honeysuckle extract, centella asiatica extract, early celery extract, iris extract, oyster shell extract, calcium chloride mixed treatment into the sponge bone needle, and the wheat oligopeptide is grafted on the outside of the sponge bone needle to obtain a penetration composition:

[0031] Wheat oligopeptide can effectively inhibit the overexpression of inflammatory factors and reduce inflammatory reactions. Its small molecule feature can penetrate into the dermis layer of the skin, directly act on the cells at the inflammatory site, regulate immune response, reduce the sensitive state of the skin, protect cells from oxidative stress damage, promote collagen synthesis, and help repair damaged skin tissue.

[0032] When the sponge bone needle pierces into the skin, calcium ions are released, the concentration of calcium ions in the skin cells is increased, the macrophages at the acne inflammatory skin site are activated to synthesize low-concentration nitric oxide from arginine and other various amino acids contained in the andrias davidianus collagen, repair the inflammatory acne, and promote wound healing; meanwhile, the synergistic effect of the pomegranate seed extract, the honeysuckle extract, the gynura japonica extract, the early celery extract, the iris extract and the oyster shell extract further repairs the skin;

[0033] Further, the chlorogenic acid in the honeysuckle extract, the ellagic acid in the pomegranate seed extract and the flavonoid components in the iris extract inhibit the propionibacterium acnes from different angles: the chlorogenic acid destroys the bacterial cell membrane, the ellagic acid inhibits the bacterial metabolic enzyme activity, and the iris flavone interferes with the bacterial reproductive cycle, the three components synergistically enhance the antibacterial effect and reduce the inflammatory stimulation caused by bacteria; the zinc element in the oyster shell extract further assists in inhibiting the growth of bacteria and simultaneously regulates the skin microecological balance; the iris extract and the gynura japonica extract synergistically regulate keratin metabolism, promote normal keratin shedding, inhibit excessive proliferation of keratinocytes, avoid blockage of the hair follicle orifice, the early celery extract reduces sebum secretion, improves the oil environment in the hair follicle, and improves and repairs the skin inflammatory acne problem through the synergistic effect of the components. BRIEF DESCRIPTION OF DRAWINGS

[0034] The application will be further described below in conjunction with the drawings.

[0035] Figure 1 is a scanning electron microscope magnified view of the sponge bone needle morphology of preparation example 2 of the application;

[0036] Figure 2 is a Fourier transform infrared (FTIR) spectrum of the product obtained in different steps in the preparation process of example 2 of the application. DETAILED DESCRIPTION

[0037] The technical solutions in the embodiments of the application will be clearly and completely described below in conjunction with the drawings of the embodiments of the application. Obviously, the described embodiments are only part of the embodiments of the application, rather than all the embodiments of the application. Based on the embodiments in the application, all other embodiments obtained by those of ordinary skill in the art without creative labor fall within the protection scope of the application.

[0038] Preparation example 1

[0039] A preparation method of high-purity sponge bone needles, comprising the following steps:

[0040] S1, cut the natural sponge into pieces, soak in deionized water, and ultrasonically oscillate at a frequency of 35 Hz for 1 h, filter, and wash the sponge pieces with deionized water 4 times to obtain pretreated sponge;

[0041] S2, the pretreated sponge is taken into an extraction tank, deionized water and alkaline protease are added, a sodium hydroxide-sodium hexametaphosphate solution is added to adjust pH to 10, enzymolysis is carried out at 40 DEG C for 4 h, then enzyme is inactivated at 110 DEG C for 10 min, filtration is carried out, the solid phase is washed to neutral, drying is carried out, and a first enzymolysis product is obtained; the mass ratio of the pretreated sponge, deionized water and alkaline protease is 1:100:0.02; the sodium hydroxide-sodium hexametaphosphate solution is prepared by uniformly mixing 4 g of sodium hydroxide, 1.8 g of sodium hexametaphosphate and 100 mL of deionized water;

[0042] S3, the first enzymolysis product is taken into an extraction tank, deionized water and ginger protease are added, a 1 mol / L hydrochloric acid solution is added to adjust pH to 4.5, enzymolysis is carried out at 40 DEG C for 2 h, then enzyme is inactivated at 110 DEG C for 10 min, filtration is carried out, the solid phase is washed to neutral, a second enzymolysis product is obtained, vacuum freeze drying is carried out at-80 DEG C for 6 h to obtain a loose powder, and the high-purity sponge bone needle is obtained; the mass ratio of the first enzymolysis product, deionized water and ginger protease is 1:100:0.0075.

[0043] Preparation Example 2

[0044] A preparation method of a high-purity sponge bone needle, comprising the following steps:

[0045] S1, natural sponge is cut into pieces, soaked in deionized water, and subjected to ultrasonic oscillation at 40 Hz for 1.5 h, filtration is carried out, the sponge pieces are washed with deionized water for 5 times, and pretreated sponge is obtained;

[0046] S2, the pretreated sponge is taken into an extraction tank, deionized water and alkaline protease are added, a sodium hydroxide-sodium hexametaphosphate solution is added to adjust pH to 10, enzymolysis is carried out at 40 DEG C for 4 h, then enzyme is inactivated at 110 DEG C for 10 min, filtration is carried out, the solid phase is washed to neutral, drying is carried out, and a first enzymolysis product is obtained; the mass ratio of the pretreated sponge, deionized water and alkaline protease is 1:100:0.02; the sodium hydroxide-sodium hexametaphosphate solution is prepared by uniformly mixing 4 g of sodium hydroxide, 1.8 g of sodium hexametaphosphate and 100 mL of deionized water;

[0047] S3, the first enzymolysis product is taken into an extraction tank, deionized water and ginger protease are added, a 1 mol / L hydrochloric acid solution is added to adjust pH to 4.5, enzymolysis is carried out at 40 DEG C for 2 h, then enzyme is inactivated at 110 DEG C for 10 min, filtration is carried out, the solid phase is washed to neutral, a second enzymolysis product is obtained, vacuum freeze drying is carried out at-80 DEG C for 6 h to obtain a loose powder, and the high-purity sponge bone needle is obtained; the mass ratio of the first enzymolysis product, deionized water and ginger protease is 1:100:0.0075.

[0048] As shown in the accompanying Figure 1As shown, the scanning electron microscope of the sponge bone needle prepared in the embodiment is smooth in surface and complete in structure.

[0049] Preparation Example 3

[0050] A preparation method of high-purity sponge bone needles comprises the following steps:

[0051] S1, cut natural sponges, soak in deionized water, and ultrasonically oscillate at 45 Hz for 2 h, filter, and wash the sponge fragments with deionized water for 5 times to obtain pretreated sponges;

[0052] S2, place the pretreated sponges in an extraction tank, add deionized water and alkaline protease, add a sodium hydroxide-sodium hexametaphosphate solution to adjust the pH to 10, and enzymatically hydrolyze at 40℃ for 6 h, then inactivate the enzyme at 110℃ for 10 min, filter, wash the solid phase to neutral, and dry to obtain a first enzymatic hydrolysate; the mass ratio of the pretreated sponges, deionized water, and alkaline protease is 1.5:100:0.04; the sodium hydroxide-sodium hexametaphosphate solution is prepared by uniformly mixing and stirring 8 g of sodium hydroxide, 2.2 g of sodium hexametaphosphate, and 100 mL of deionized water;

[0053] S3, place the first enzymatic hydrolysate in an extraction tank, add deionized water and ginger protease, add a 1 mol / L hydrochloric acid solution to adjust the pH to 4.5, and enzymatically hydrolyze at 40℃ for 3 h, then inactivate the enzyme at 110℃ for 10 min, filter, wash the solid phase to neutral to obtain a second enzymatic hydrolysate, and vacuum freeze-dry at -80℃ for 10 h to obtain a loose powder, which is the high-purity sponge bone needles; the mass ratio of the first enzymatic hydrolysate, deionized water, and ginger protease is 1.2:100:0.014.

[0054] Preparation Example 4

[0055] A preparation method of high-purity sponge bone needles comprises the following steps:

[0056] S1, cut natural sponges, soak in deionized water, and ultrasonically oscillate at 45 Hz for 2 h, filter, and wash the sponge fragments with deionized water for 5 times to obtain pretreated sponges;

[0057] S2, the pretreated sponges are placed in an extraction tank, deionized water and alkaline protease are added, a sodium hydroxide-sodium hexametaphosphate solution is added to adjust the pH to 10, and enzymolysis is carried out at 40℃ for 5h, and then the enzyme is inactivated at 110℃ for 10min, filtration is performed, the solid phase is washed to neutral, and vacuum freeze-drying is performed at -80℃ for 8h to obtain loose powder, and the high-purity spongy bone needles are obtained; the mass ratio of the pretreated sponges, deionized water and alkaline protease is 1.25:100:0.03; the sodium hydroxide-sodium hexametaphosphate solution is prepared by uniformly mixing 6g of sodium hydroxide, 2.0g of sodium hexametaphosphate and 100mL of deionized water.

[0058] The spongy bone needles prepared in Preparation Examples 1-4 are taken to detect the content of silicon dioxide, as shown in Table 1 below.

[0059] Table 1

[0060]

[0061]

[0062] As can be seen from Table 1, the high-purity spongy bone needles are obtained by twice enzymolysis in Preparation Examples 1-3; and the purity is reduced due to only one enzymolysis in Preparation Example 4.

[0063] Example 1

[0064] The spongy bone needles prepared in Preparation Example 2 are used to prepare a penetration promoting composition for repairing skin inflammation and acne, and the preparation method of the penetration promoting composition comprises the following steps:

[0065] A1, the giant salamander skin collagen, pomegranate seed extract, honeysuckle extract, centella asiatica extract, early celery extract, iris extract, oyster shell extract, anhydrous ethanol, deionized water and calcium chloride are mixed, and ultrasonic stirring is performed for 15min to obtain material A; the mass ratio of the giant salamander skin collagen, pomegranate seed extract, honeysuckle extract, centella asiatica extract, early celery extract, iris extract, oyster shell extract, anhydrous ethanol, deionized water and calcium chloride is 25:8:4:18:4:2:3:40:60:0.4;

[0066] A2, the high-purity spongy bone needles, N,N-dimethylformamide and deionized water are mixed, and ultrasonic dispersion is performed for 30min, and then succinic anhydride and 3-aminopropyl triethoxysilane are added, and heating and stirring are performed in a water bath at 50℃ for 5h, and then centrifugal separation is performed, the solid phase is washed, and vacuum freeze-drying is performed to obtain material B; the mass ratio of the high-purity spongy bone needles, N,N-dimethylformamide, deionized water, succinic anhydride and 3-aminopropyl triethoxysilane is 0.6g:2mL:30mL:4g:8g;

[0067] A3, take the material B, wheat oligopeptide and deionized water mixed, ultrasonic treatment 20 min, centrifugal separation, take the solid phase washing, drying, get material C; the material B, wheat oligopeptide and deionized water's proportioning ratio is 0.4g:10mg:10mL;

[0068] A4, take the material C, material A mixed, ultrasonic dispersion 30 min, in 0.1MPa's vacuum condition pressure maintaining 30 min, filter, take solid phase and get material D; the material C, material A's mass ratio is 1:10;

[0069] A5, take the aluminum glycyrrhizinate, tartaric acid, deionized water mixed, stirring 20 min, join polyacrylic acid sodium, continue stirring 30 min, join the material D, continue stirring 5 min, get the penetration composition; the aluminum glycyryrrhizinate, tartaric acid, deionized water, polyacrylic acid sodium, material D's proportioning ratio is 0.3g:0.4g:50mL:8g:1.5g.

[0070] Example 2

[0071] The preparation method of the penetration composition prepared by the sponge bone needle obtained in preparation example 2 for repairing skin inflammation acne includes the following steps:

[0072] A1, take the giant salamander skin collagen, pomegranate seed extract, honeysuckle extract, gynura extract, early celery extract, iris extract, oyster shell extract, anhydrous ethanol, deionized water, calcium chloride mixed, ultrasonic stirring 20 min, get material A; the giant salamander skin collagen, pomegranate seed extract, honeysuckle extract, gynura extract, early celery extract, iris extract, oyster shell extract, anhydrous ethanol, deionized water's mass ratio is 30:9:5:22:5:3:4:45:70:0.6;

[0073] A2, take the high-purity sponge bone needle, N,N-dimethylformamide, deionized water mixed, ultrasonic dispersion 35 min, join succinic anhydride and 3-aminopropyl triethoxysilane, in 55 ℃ water bath heating stirring 6h, centrifugal separation, take the solid phase washing, vacuum freeze drying, get material B; the high-purity sponge bone needle, N,N-dimethylformamide, deionized water, succinic anhydride and 3-aminopropyl triethoxysilane's proportioning ratio is 0.7g:2.5mL:30mL:4.5g:9g;

[0074] A3, take the material B, wheat oligopeptide and deionized water mixed, ultrasonic treatment 25 min, centrifugal separation, take the solid phase washing, drying, get material C; the material B, wheat oligopeptide and deionized water's proportioning ratio is 0.6g:12mg:12mL;

[0075] A4, take the material C, material A mixed, ultrasonic dispersion 35 min, in the vacuum condition of 0.1 MPa pressure maintaining 35 min, filtration, take solid phase to obtain material D, the mass ratio of the material C, material A is 1:10;

[0076] A5, take the ganxu aluminum, tartaric acid, deionized water mixed, stirring 25 min, add polyacrylic acid sodium, continue to stir 35 min, add the material D, continue to stir 8 min, obtain the penetration enhancer composition, the ganxu aluminum, tartaric acid, deionized water, polyacrylic acid sodium, material D is 0.4 g:0.5 g:50 mL:9 g:1.8 g.

[0077] As shown in the preparation process of the embodiment, the Fourier transform infrared (FTIR) spectrum of the product obtained in different steps can indicate that the sponge bone needle surface is successfully grafted with wheat oligopeptide. Figure 2

[0078] Example 3

[0079] The sponge bone needle obtained in preparation example 2 is used to prepare a penetration enhancer composition for repairing skin inflammation acne, and the preparation method of the penetration enhancer composition comprises the following steps:

[0080] A1, take the giant salamander skin collagen, pomegranate seed extract, honeysuckle extract, gynura extract, early celery extract, iris extract, oyster shell extract, anhydrous ethanol, deionized water, calcium chloride mixed, ultrasonic stirring 25 min, obtain material A, the mass ratio of the giant salamander skin collagen, pomegranate seed extract, honeysuckle extract, gynura extract, early celery extract, iris extract, oyster shell extract, anhydrous ethanol, deionized water is 35:10:6:26:6:4:5:50:80:0.8;

[0081] A2, take the high-purity sponge bone needle, N,N-dimethylformamide, deionized water mixed, ultrasonic dispersion 40 min, add succinic anhydride and 3-aminopropyl triethoxysilane, heat stirring at 60°C water bath for 7h, centrifugal separation, take the solid phase washing, vacuum freeze drying, obtain material B, the high-purity sponge bone needle, N,N-dimethylformamide, deionized water, succinic anhydride and 3-aminopropyl triethoxysilane is 0.8 g:3 mL:30 mL:5 g:10 g;

[0082] A3, take the material B, wheat oligopeptide and deionized water mixed, ultrasonic treatment 30 min, centrifugal separation, take the solid phase washing, drying, obtain material C, the material B, wheat oligopeptide and deionized water is 0.8 g:15 mg:15 mL;

[0083] ​A4, taking the material C, material A mixed, ultrasonic dispersion 40 min, in the vacuum condition of 0.1 MPa pressure maintaining 40 min, filtration, take solid phase to obtain material D; the mass ratio of the material C, material A is 1:10;

[0084] A5, taking the material D, continue to stir 10 min, the penetration enhancer composition is obtained; the amount ratio of the ganhydroxyaluminum, tartaric acid, deionized water, polyacrylic acid sodium, material D is 0.5g:0.6g:50mL:10g:2.2g.

[0085] Comparative Example 1

[0086] The comparative example provides a penetration enhancer composition, which is only different from example 2 in that the component does not contain big skink skin collagen, and the reduced amount is distributed to pomegranate seed extract, honeysuckle extract, centella asiatica extract, early celery extract, iris extract, oyster shell extract, calcium chloride in the ratio of parts, and the rest of the components and the amount remain unchanged.

[0087] Comparative Example 2

[0088] The comparative example provides a penetration enhancer composition, which is only different from example 2 in that the component does not contain pomegranate seed extract, and the reduced amount is distributed to big skink skin collagen, honeysuckle extract, centella asiatica extract, early celery extract, iris extract, oyster shell extract, calcium chloride in the ratio of parts, and the rest of the components and the amount remain unchanged.

[0089] Comparative Example 3

[0090] The comparative example provides a penetration enhancer composition, which is only different from example 2 in that the component does not contain honeysuckle extract, and the reduced amount is distributed to big skink skin collagen, pomegranate seed extract, centella asiatica extract, early celery extract, iris extract, oyster shell extract, calcium chloride in the ratio of parts, and the rest of the components and the amount remain unchanged.

[0091] Comparative Example 4

[0092] The comparative example provides a penetration enhancer composition, which is only different from example 2 in that the component does not contain centella asiatica extract, and the reduced amount is distributed to big skink skin collagen, pomegranate seed extract, honeysuckle extract, early celery extract, iris extract, oyster shell extract, calcium chloride in the ratio of parts, and the rest of the components and the amount remain unchanged.

[0093] Comparative Example 5

[0094] The comparative example provides a penetration enhancing composition, which is only different from example 2 in that the component does not contain early celery extract, and the reduced amount is proportionally distributed to andrews skin collagen, pomegranate seed extract, honeysuckle extract, centella asiatica extract, iris extract, oyster shell extract, calcium chloride, and the rest of the components and the amount remain unchanged.

[0095] Comparative example 6

[0096] The comparative example provides a penetration enhancing composition, which is only different from example 2 in that the component does not contain early celery extract, and the reduced amount is proportionally distributed to andrews skin collagen, pomegranate seed extract, honeysuckle extract, centella asiatica extract, iris extract, oyster shell extract, calcium chloride, and the rest of the components and the amount remain unchanged.

[0097] Comparative example 7

[0098] The comparative example provides a penetration enhancing composition, which is only different from example 2 in that the component does not contain early celery extract, and the reduced amount is proportionally distributed to andrews skin collagen, pomegranate seed extract, honeysuckle extract, centella asiatica extract, iris extract, oyster shell extract, calcium chloride, and the rest of the components and the amount remain unchanged.

[0099] Comparative example 8

[0100] The comparative example provides a penetration enhancing composition, which is only different from example 2 in that the component does not contain early celery extract, and the reduced amount is proportionally distributed to andrews skin collagen, pomegranate seed extract, honeysuckle extract, centella asiatica extract, iris extract, oyster shell extract, calcium chloride, and the rest of the components and the amount remain unchanged.

[0101] Comparative example 9

[0102] The difference from example 2 is that the preparation method of the penetration enhancing composition comprises the following steps:

[0103] A1, take andrews skin collagen, pomegranate seed extract, honeysuckle extract, centella asiatica extract, early celery extract, iris extract, oyster shell extract, anhydrous ethanol, deionized water, calcium chloride, mix, ultrasonic stirring for 20 min, get material A;

[0104] A2, take the high-purity sponge bone needle, material A, mix, ultrasonic dispersion for 35 min, keep pressure under the condition of 0.1 MPa vacuum for 35 min, filter, take the solid phase to get material D; the mass ratio of the high-purity sponge bone needle and material A is 1:10;

[0105] A5, take gallium hydroxy aluminum, tartaric acid, deionized water mixed, stirring 25 min, add polyacrylic acid sodium, continue to stir 35 min, add the material D, continue to stir 8 min, the penetration composition is obtained; the gallium hydroxy aluminum, tartaric acid, deionized water, polyacrylic acid sodium, material D is 0.4g: 0.5g: 50ml: 9g: 1.8g.

[0106] Comparative Example 10

[0107] The difference from Example 2 is that the preparation method of the penetration composition comprises the following steps:

[0108] A1, take Megalobatrachus skin collagen, pomegranate seed extract, honeysuckle extract, Centella asiatica extract, early celery extract, iris extract, oyster shell extract, anhydrous ethanol, deionized water, calcium chloride mixed, ultrasonic stirring 20 min, to get material A;

[0109] A2, take gallium hydroxy aluminum, tartaric acid, deionized water mixed, stirring 25 min, add polyacrylic acid sodium, continue to stir 35 min, add the material A, continue to stir 8 min, the penetration composition is obtained; the gallium hydroxy aluminum, tartaric acid, deionized water, polyacrylic acid sodium, material A is 0.4g: 0.5g: 50ml: 9g: 10g.

[0110] Performance test

[0111] Test Example 1

[0112] Select 130 six-week-old ICR Mice mice, randomly divided into 13 groups, back skin shaving 2cm×2cm, the 2-4th day after shaving, inject Propionibacterium acnes (0.2×10 7 CFU / mL) on the back skin of mice, construct mouse acne model, start pressing and applying the penetration composition prepared by Example 1-Example 3 and Comparative Example 1-Comparative Example 10 on the 5th day, continue for 14 days, compare the acne area before and after application, as shown in Table 2 below.

[0113] Table 2

[0114]

[0115] As shown in Table 2, the penetration composition prepared by Example 1-Example 3 has good effect on treating skin acne; Comparative Example 1-Comparative Example 8 lacks the synergistic effect among Megalobatrachus skin collagen, pomegranate seed extract, honeysuckle extract, Centella asiatica extract, early celery extract, iris extract, oyster shell extract and calcium chloride, resulting in reduced treatment effect; Comparative Example 9 does not modify the surface of the sponge bone needle, resulting in reduced treatment effect; Comparative Example 10 does not add sponge bone needle, resulting in poor penetration effect and greatly reduced treatment effect.

[0116] Test Example 2 Volunteer Treatment Effect

[0117] Test Subjects: Randomly selected volunteers with different degrees of acne inflammation, 130 people, aged 26-48 years old, among them, 68 people with neutral skin, 41 people with oily skin, and 21 people with dry skin. 10 people were divided into 13 groups, and each group corresponded to one of the penetration enhancing compositions in the examples and comparative examples. The penetration enhancing compositions prepared in Examples 1-3 and Comparative Examples 1-10 were applied to the acne inflammation sites of the patients.

[0118] Test Method: 2g of the penetration enhancing compositions prepared in Examples 1-3 and Comparative Examples 1-10 were applied to the acne inflammation sites and within a radius of 3cm, and were applied twice a day, morning and evening. After two weeks of continuous use, the effect of the penetration enhancing compositions was evaluated, and the test results are shown in Table 3.

[0119] Evaluation Criteria for Effect of Use:

[0120] Grade I: Significant repair effect, significant improvement in acne sites;

[0121] Grade II: Better repair effect, more significant improvement in acne sites;

[0122] Grade III: General repair effect, no significant improvement in acne sites;

[0123] Grade IV: Poor repair effect, no improvement in acne sites.

[0124] Table 3

[0125]

[0126]

[0127] As shown in Table 3, the penetration enhancing compositions of Examples 1-3 are considered to have better repair effect; while the soothing and repair effects of the penetration enhancing compositions of Comparative Examples 1-9 are worse than those of Examples 1-3, and Comparative Example 10 is considered to have very poor effect.

[0128] The above describes some embodiments of the present application in detail, but the content described is only the preferred embodiments of the present application, and cannot be considered to limit the scope of the implementation of the present application. Any equivalent changes and improvements made in accordance with the scope of the present application should still be within the scope of the present patent.

Claims

1. A method for preparing high-purity sponge spicules, characterized in that: The following steps are involved: S1. Cut a natural sponge into pieces, soak it in deionized water, and ultrasonically vibrate it at a frequency of 35-45 Hz for 1-2 hours. Filter the pieces, and wash the sponge pieces with deionized water 4-5 times to obtain a pretreated sponge. S2. Place the pretreated sponge in an extraction tank, add deionized water and alkaline protease, adjust the pH to 10, perform enzymatic hydrolysis at 40°C for 4-6 hours, then inactivate the enzyme at 110°C for 10 minutes, filter, wash the solid phase, and dry to obtain a primary hydrolyzate; S3. Place the primary hydrolyzate in an extraction tank, add deionized water and ginger protease, adjust the pH to 4.5, hydrolyze at 40°C for 2-3 hours, inactivate the enzyme at 110°C for 10 minutes, filter, wash the solid phase, obtain a secondary hydrolyzate, and freeze-dry at -80°C in a vacuum freeze-drying process for 6-10 hours until the product becomes a loose powder, thereby obtaining the high-purity sponge spicule.

2. The method for preparing a high-purity sponge spicule and its penetration-promoting application according to claim 1, characterized in that: In step S2, the mass ratio of the pretreated sponge, deionized water and alkaline protease is 1-1.5:100:0.02-0.

04.

3. The method for preparing a high-purity sponge spicule and its penetration-promoting application according to claim 1, characterized in that: In step S3, the mass ratio of the primary enzymatic hydrolysate, deionized water and ginger protease is 1-1.2:100:0.0075-0.

014.

4. A high-purity sponge spicule prepared by the preparation method according to any one of claims 1 to 3.

5. The application of the high-purity sponge spicule for promoting penetration according to claim 4, characterized in that: A penetration-promoting composition is prepared using sponge spicules for repairing skin inflammation and acne. The preparation method of the penetration-promoting composition comprises the following steps: A1. Mix giant salamander skin collagen, pomegranate seed extract, honeysuckle extract, Centella asiatica extract, celery extract, iris extract, oyster shell extract, anhydrous ethanol, deionized water, and calcium chloride, and stir ultrasonically for 15-25 minutes to obtain material A; A2. Mix the high-purity sponge spicules, N,N-dimethylformamide, and deionized water, and ultrasonically disperse for 30-40 min. Add succinic anhydride and 3-aminopropyltriethoxysilane, heat in a water bath at 50-60° C. with stirring for 5-7 h, centrifuge, wash the solid phase, and freeze-dry in vacuo to obtain material B. A3, taking the material B, wheat oligopeptide and deionized water, mixing them, ultrasonically treating them for 20-30 minutes, centrifuging them, washing the solid phase, and drying them to obtain material C; A4, take the material C and material A, mix them, ultrasonically disperse them for 30-40 minutes, maintain the pressure under 0.1 MPa vacuum conditions for 30-40 minutes, filter, and take the solid phase to obtain material D; A5. Mix aluminum glycolate, tartaric acid, and deionized water, stir for 20-30 minutes, add sodium polyacrylate, continue stirring for 30-40 minutes, add material D, and continue stirring for 5-10 minutes to obtain the penetration-enhancing composition.

6. The application of the high-purity sponge spicule for promoting penetration according to claim 5, characterized in that: In step A1, the mass ratio of the giant salamander skin collagen, pomegranate seed extract, honeysuckle extract, Centella asiatica extract, celery extract, iris extract, oyster shell extract, anhydrous ethanol, and deionized water is 25-35:8-10:4-6:18-26:4-6:2-4:3-5:40-50:60-80:0.4-0.

8.

7. The application of the high-purity sponge spicule for promoting penetration according to claim 5, characterized in that: In step A2, the ratio of the high-purity sponge spicule, N,N-dimethylformamide, deionized water, succinic anhydride and 3-aminopropyltriethoxysilane is 0.6-0.8 g: 2-3 mL: 30 mL: 4-5 g: 8-10 g.

8. The application of the high-purity sponge spicule for promoting penetration according to claim 5, characterized in that: In step A3, the dosage ratio of material B, wheat oligopeptide and deionized water is 0.4-0.8 g:10-15 mg:10-15 mL.

9. The application of a high-purity sponge spicule for promoting penetration according to claim 5, characterized in that: In step A4, the mass ratio of material C to material A is 1:

10.

10. The application of the high-purity sponge spicule for promoting penetration according to claim 5, characterized in that: In step A5, the dosage ratio of aluminum glycolate, tartaric acid, deionized water, sodium polyacrylate, and material D is 0.3-0.5 g: 0.4-0.6 g: 50 mL: 8-10 g: 1.5-2.2 g.

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