Nano plaster and preparation method thereof

Abstract

The invention relates to the field of medicine emplastrum, and discloses a nanometer emplastrum and a preparation method thereof.The nanometer emplastrum comprises 55%-60% of nanometer particles and 40%-45% of traditional Chinese medicine active ingredients; the nano-particles comprise 25%-30% of animal protein extract, 15%-20% of mineral protein extract and 50%-55% of a nano-carrier base material, the animal protein extract comprises fish scale collagen; the mineral protein extract comprises diatomite protein; the traditional Chinese medicine active components comprise 12%-15% of an agaric polysaccharide extract, 10%-15% of a panax notoginseng saponins extract, 5%-10% of a safflower flavone extract, 8%-12% of a salvianolic acid extract and 5%-10% of malic acid. According to the invention, a porous nano-carrier constructed by animal protein and mineral protein breaks through the bottleneck of transdermal efficiency of a traditional plaster, and based on the characteristic of high specific surface area of fish scale collagen and diatomite protein composite nanoparticles and in combination with the pH regulation effect of malic acid, the transdermal permeability of macromolecular active components such as auricularia auricula polysaccharide, notoginsenoside and the like is remarkably improved.

Description

Technical Field

[0001] The invention relates to the technical field of drug plasters, in particular to a nano plaster and a preparation method thereof. Background Art

[0002] As an important carrier of traditional external treatment, Chinese herbal transdermal patches belong to the field of external transdermal therapeutic preparations and are mainly used for local treatment of symptoms such as bruises, rheumatic pain and muscle strain.

[0003] Existing Chinese medicine plasters usually extract and concentrate the medicinal materials with ethanol, mix them with a matrix (such as vaseline, beeswax), and add a transdermal enhancer (such as azone) to make a paste, which is then applied to non-woven fabrics or stretch fabrics to fit the affected area. Its mechanism of action relies on the gradual diffusion of Chinese medicine ingredients (such as Panax notoginseng total saponins to promote microcirculation and salvianolic acid to inhibit inflammatory factors) through the stratum corneum of the skin to the subcutaneous tissue, combined with the warming effect of physical application to achieve local blood flow improvement and pain relief. Typical products include blood-activating patches with added resin ingredients such as frankincense and myrrh, as well as cooling analgesic patches containing borneol and menthol.

[0004] Although traditional Chinese medicine patches have the advantages of high safety and few side effects, their efficacy is limited by the following technical bottlenecks: Low transdermal efficiency: The active ingredients (such as polysaccharides and saponins) have a large molecular weight and strong polarity, making it difficult to effectively penetrate the skin's stratum corneum barrier, resulting in a transdermal absorption rate generally lower than 30% and insufficient blood drug concentration in deep tissues; Insufficient sustained-release performance: Traditional matrices (such as oils and colloids) have poor drug loading and controlled-release capabilities. Most patches release more than 80% of the active ingredients within 6-8 hours, and cannot achieve long-term sustained release, requiring frequent replacement; Irritation risk: Some penetration enhancers (such as high-concentration ethanol) are prone to cause skin allergic reactions, and residual plant fibers or particles of the drug may clog pores, exacerbating problems such as redness, swelling, and itching. Summary of the invention

[0005] In view of the shortcomings of the prior art, the present invention provides a nano plaster and a preparation method thereof, which solves the problem that the existing plasters for treating injuries are mainly made of Chinese medicinal materials. Its efficacy is to promote blood circulation and remove blood stasis, and eliminate swelling and pain. However, there are problems such as poor permeability to the skin, slow drug properties, and low efficacy.

[0006] To achieve the above objectives, the present invention is implemented through the following technical scheme: a nano patch, comprising 55%-60% nanoparticles and 40%-45% active ingredients of traditional Chinese medicine; the nanoparticles include: 25%-30% animal protein extract, 15%-20% mineral protein extract, and 50%-55% nano carrier substrate.

[0007] Preferably, the animal protein extract includes: fish scale collagen.

[0008] Preferably, the mineral protein extract comprises: diatomaceous earth protein.

[0009] Preferably, the active ingredients of traditional Chinese medicine include: 12%-15% of Auricularia auricula polysaccharide extract, 10%-15% of Panax notoginseng total saponin extract, 5%-10% of safflower flavonoids extract, 8%-12% of salvianolic acid extract, and 5%-10% of malic acid.

[0010] A method for preparing a nano plaster comprises the following steps: S1. Extraction of active ingredients from traditional Chinese medicine: Mix Auricularia auricula, Panax notoginseng, safflower, and Salvia miltiorrhiza in a mass ratio of 3:4:2:1, crush using a traditional Chinese medicine grinder for 6-8 minutes, add 10 times the amount of 70% ethanol after sieving, place in an ultrasonic extractor for extraction 3 times, each time for 30 minutes, combine the extracts and concentrate them by rotary evaporator until there is no alcohol taste, and finally dry them in a freeze dryer for 24-36 hours to obtain a composite extract powder; S2. Extraction of animal protein and mineral protein: Add decalcified fish scales to a constant temperature enzymatic hydrolysis tank, enzymatically hydrolyze with 5% pepsin at 45°C and 120rpm for 4 hours, centrifuge for 15 minutes, dialyze the supernatant and freeze-dry to obtain high-purity collagen powder; then enzymatically hydrolyze diatomaceous earth with neutral protease at 50°C and 150rpm for 6 hours, centrifuge and dry the filtrate with a spray dryer to obtain mineral protein powder with D50≤10μm S3. Preparation of porous nanoparticles: Mix animal protein and mineral protein powder and dissolve them in deionized water, add 1% ammonium bicarbonate foaming agent, freeze in liquid nitrogen and use vacuum freeze dryer to dry for 24 hours to form a skeleton. Put the skeleton into nano ball mill for 2 hours, pass through 200 mesh sieve to obtain 80-120nm porous nanoparticles; S4. Active ingredient loading: The composite extract and malic acid were dissolved in pH 5.5 citric acid buffer, added to porous nanoparticles, adsorbed in a constant temperature oscillator for 8 hours, centrifuged and dried in a vacuum drying oven for 6 hours, and the final water content was controlled to be ≤3%; S5. Paste molding and patch preparation: Mix sodium carboxymethyl cellulose, glycerol, azone and deionized water, homogenize with a high shear homogenizer for 10 minutes to form a gel, use a coater to coat the film, then cut into 5×7 cm patches and cover with release paper for packaging.

[0011] The present invention provides a nano plaster and a preparation method thereof, which has the following beneficial effects: 1. The porous nanocarrier constructed by animal protein and mineral protein in the present invention breaks through the bottleneck of transdermal efficiency of traditional plasters. Based on the high specific surface area characteristics of fish scale collagen and diatomaceous earth protein composite nanoparticles and the pH regulating effect of malic acid, the transdermal permeability of macromolecular active ingredients such as auricularia auricula polysaccharide and notoginseng saponin is significantly improved.

[0012] 2. The present invention uses a biocompatible material system to reduce the risk of skin irritation, and has both antibacterial and repair functions. It uses a nano-skeleton formed by high-purity collagen extracted by pepsin-directed enzymatic hydrolysis and diatomaceous earth mineral protein, which has no fiber residue and uniform pores to avoid pore blockage; at the same time, the natural antibacterial properties of the protein matrix reduce the use of additional preservatives.

[0013] 3. The present invention realizes efficient loading and stable drug release of complex active ingredients through freeze-drying-ball milling preparation process, combines the through-channels formed by liquid nitrogen quick freezing with nano-ball milling to precisely control the particle size, so that the loading rate of ingredients such as salvianolic acid and safflower flavonoids is increased to more than 95%; the directional adsorption of citric acid buffer further optimizes the uniformity of drug distribution. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 It is a flow chart of the preparation method of the present invention. DETAILED DESCRIPTION

[0015] The following will be combined with the drawings in the specification of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0016] Embodiment 1: Please refer to the attached Figure 1 , the embodiment of the present invention provides a nano plaster, including 55%-60% nanoparticles and 40%-45% active ingredients of traditional Chinese medicine; The nanoparticles include: 25%-30% animal protein extract, 15%-20% mineral protein extract, and 50%-55% nanocarrier substrate; The animal protein extract includes: fish scale collagen; The mineral protein extract comprises: diatomaceous earth protein; The active ingredients of the traditional Chinese medicine include: 12%-15% of auricularia auricula polysaccharide extract, 10%-15% of Panax notoginseng total saponin extract, 5%-10% of safflower flavonoids extract, 8%-12% of salvianolic acid extract, and 5%-10% of malic acid.

[0017] Beneficial effects of Example 1: In this Example 1, the nanoparticle substrate constructed by fish scale collagen and diatomaceous earth protein cooperates with active ingredients of traditional Chinese medicine such as black fungus polysaccharide and Panax notoginseng saponin to achieve efficient sustained release and transdermal absorption; the malic acid in the basic formula regulates the pH value of body fluids and enhances the stability of ingredients. It is suitable for long-term repair of chronic pain (such as neck and shoulder pain), has no obvious irritation, and has a skin adaptability of more than 95%.

[0018] A method for preparing a nano plaster comprises the following steps: S1. Extraction of active ingredients from traditional Chinese medicine: Auricularia auricula, Panax notoginseng, safflower and Salvia miltiorrhiza were mixed in a mass ratio of 3:4:2:1, and crushed for 6-8 minutes using a traditional Chinese medicine grinder (rotation speed 25000rpm, mesh size 0.25mm), and then sieved and added with 10 times the amount of 70% ethanol, and then placed in an ultrasonic extractor (40kHz, 300W, 50℃) for extraction for 3 times, each time for 30 minutes, and after combining the extracts, concentrated by a rotary evaporator (100rpm, 50℃ water bath, vacuum 0.098MPa) until there was no alcohol taste, and finally dried by a freeze dryer (-50℃ cold trap, vacuum 5Pa) for 24-36 hours to obtain a composite extract powder; S2. Extraction of animal protein and mineral protein: Add decalcified fish scales to a constant temperature enzymatic hydrolysis tank, enzymatically hydrolyze with 5% pepsin (pH=2.0) at 45℃, 120rpm for 4 hours, centrifuge (8000rpm, 4℃, 15 minutes), dialyze the supernatant and freeze-dry to obtain high-purity collagen powder; then enzymatically hydrolyze diatomaceous earth with neutral protease (pH=7.0) at 50℃, 150rpm for 6 hours, centrifuge (6000rpm), and dry the filtrate in a spray dryer (inlet air 180℃, atomizing disk 18000rpm) to obtain mineral protein powder with D50≤10μm S3. Preparation of porous nanoparticles: animal protein and mineral protein powder (3:2) were mixed and dissolved in deionized water, 1% ammonium bicarbonate foaming agent was added, and the skeleton was formed by quick freezing in liquid nitrogen and drying in a vacuum freeze dryer (-80℃ cold trap, vacuum 5Pa) for 24 hours. The skeleton was put into a nano ball mill (revolution 300rpm, rotation 600rpm) for 2 hours (zirconia balls, ball-to-material ratio 10:1), and passed through a 200 mesh sieve to obtain 80-120nm porous nanoparticles; S4. Active ingredient loading: The composite extract and malic acid were dissolved in pH 5.5 citric acid buffer (concentration 20%), added to porous nanoparticles, adsorbed in a constant temperature oscillator (45°C, 150 times / min) for 8 hours, centrifuged and placed in a vacuum drying oven (40°C, vacuum -0.09MPa) for 6 hours, and the final water content was controlled to be ≤3%; S5. Paste molding and patch preparation: Mix sodium carboxymethyl cellulose, glycerol, azone and deionized water, and homogenize them with a high shear homogenizer (5000rpm) for 10 minutes to form a gel. Use a coater (coating thickness 0.2mm, drying 50℃, speed 10m / min) to coat the paste into a film, then cut it into 5×7cm patches and cover them with release paper for packaging.

[0019] Embodiment 2: A nanopatch, comprising 52%-58% nanoparticles and 42%-48% active ingredients of traditional Chinese medicine; The nanoparticles include: 25%-30% animal protein extract, 15%-20% mineral protein extract, and 50%-55% nanocarrier substrate; The animal protein extract includes: chitin-modified shrimp shell collagen; The mineral protein extract comprises: volcanic ash silicate protein; The active ingredients of the traditional Chinese medicine include: 10%-13% of auricularia auricula polysaccharide extract, 5%-8% of Panax notoginseng total saponin extract, 6%-9% of safflower flavonoids extract, 7%-10% of salvianolic acid extract, 5%-10% of malic acid, and 8%-12% of asiatica glycoside extract; Beneficial effects of Example 2: In Example 2, chitin is used to modify shrimp shell collagen and volcanic ash silicate protein to enhance the adsorption and antibacterial properties of nanoparticles, and the release of inflammatory factors is inhibited by combining with Centella asiatica extract, which significantly improves the targeted repair efficiency of the patch for skin inflammation (such as eczema, redness and swelling); the porosity of the mineral protein is increased by 20%, the drug loading capacity is increased, and the duration of action is extended to 48 hours, with both antibacterial and epidermal regeneration functions.

[0020] Embodiment three: A nanopatch, comprising 58%-62% nanoparticles and 38%-42% active ingredients of traditional Chinese medicine; The nanoparticles include: 15%-18% animal protein extract, 25%-30% mineral protein extract, and 55%-60% nanocarrier substrate; The animal protein extract includes: fish scale collagen; The mineral protein extract comprises: diatomaceous earth protein; The active ingredients of the traditional Chinese medicine include: 3%-5% of borneol microencapsulated powder, 4%-6% of ligustrazine extract, 8%-10% of auricularia auricula polysaccharide extract, 12%-15% of Panax notoginseng total saponin extract, 10%-12% of salvianolic acid extract, and malic acid adjusted to 2%-5%; Beneficial effects of Example 3: This Example 3 takes zeolite molecular sieve mineral protein and borneol microencapsulated powder as the core to achieve directional controlled release of salvianolic acid and ligustrazine, and quickly reduce the burning sensation in the acute pain area; Panax notoginseng saponins and borneol work synergistically to enhance transdermal efficiency, and ligustrazine accelerates microcirculation, which is suitable for sports injuries or acute sprains. At the same time, after optimizing the malic acid ratio, the high temperature resistance of the paste is improved, and the shelf life at 40°C is extended to 18 months.

[0021] Comparative experiment: According to the functional emphasis of Example 1 (basic long-acting type), Example 2 (antibacterial and anti-inflammatory type), and Example 3 (fast-acting controlled-release type), the following comparative experimental scheme was designed (control group: commercially available traditional Chinese medicine plaster, without nanocarrier technology). The experimental data must be repeated at least 3 times and the average value is taken.

[0022] Experiment 1: Comparison of sustained-release performance and transdermal absorption efficiency Test objective: To verify the long-acting sustained-release characteristics of Example 1 and compare the drug release rate and skin permeation of each example. Steps and parameters: Franz Diffusion Cell Method Skin model: ex vivo pig skin (thickness 0.8 mm, maintained active after pretreatment).

[0023] Release medium: pH 7.4 phosphate buffer (37 ± 0.5 °C), rotation speed 300 rpm.

[0024] Sampling time points: 0.5h, 2h, 6h, 12h, 24h, 48h.

[0025] Test indicators: Cumulative release rate of salvianolic acid and notoginseng saponins (HPLC detection, chromatographic conditions: C18 column, mobile phase acetonitrile-0.1% phosphoric acid, flow rate 1mL / min, detection wavelength 230nm).

[0026] Control group treatment: The same dose of extract was directly applied on commercially available plaster.

[0027] Expected results: Example 1: The sustained release reaches more than 75% within 24 hours, and the release is maintained at 90% after 48 hours; Example 3: The release amount within 6 hours is ≥50% (significant rapid release effect); The release of traditional patches drops sharply after 12 hours, and only 30% to 40% remains after 48 hours.

[0028] Experiment 2: Verification of antibacterial and anti-inflammatory effects (core of Example 2) Test objective: To evaluate the ability of Example 2 to inhibit common skin pathogens and the regulation effect of inflammatory factors (IL-6, TNF-α). Steps and parameters: Antimicrobial test (disc diffusion method) Bacterial species: Staphylococcus aureus (ATCC 6538), Escherichia coli (ATCC 25922).

[0029] Culture medium: Mueller-Hinton agar, patch diameter 6 mm (containing 0.1 g of the medicinal material of the example), culture at 37°C for 24 h.

[0030] Parameters: inhibition zone diameter (mm) and minimum inhibitory concentration (MIC).

[0031] Cellular inflammation model Cell line: Human immortalized epidermal cells (HaCaT), LPS-induced inflammation.

[0032] Treatment group: intervention with the plaster extract (0.1 mg / mL) for 24 h.

[0033] Detection method: ELISA was used to determine the concentrations of IL-6 and TNF-α in the supernatant.

[0034] Expected results: Example 2 The diameter of the inhibition zone was ≥15 mm (control group <5 mm), and the MIC was reduced to 0.05 mg / mL; The IL-6 inhibition rate of Example 2 reached 70%, which was significantly higher than that of other groups (Example 1 50%, traditional plaster 30%).

[0035] Experiment 3: Acute analgesia and microcirculation improvement effects (core of Example 3) Test objective: To verify the rapid relief effect of Example 3 on the acute pain model and the ability to increase local blood flow. Steps and parameters: Rat plantar burning pain model Induction method: 40℃ hot plate (58±0.5℃), and the paw withdrawal latency (seconds) was recorded after plantar stimulation.

[0036] Drug administration group: The test was conducted 1 hour after the application of the patch, and the latency extension rate before and after drug administration was compared.

[0037] Laser Doppler blood flow imaging Model: Local muscle injury in rats (modeled by impact method).

[0038] Testing time points: 0.5h, 2h, and 6h after application, analyze the change rate of blood flow in the affected area.

[0039] Expected results: Example 3: The paw withdrawal latency is extended by ≥200% within 1 hour (Example 1: 120%, traditional plaster: 80%); In Example 3, blood flow increased to 160% of normal level 2 hours after application (in Example 2, it increased by 130%).

[0040] Experiment 4: Skin irritation and high temperature stability test Test objective: To evaluate the safety of each embodiment and the high temperature resistance of embodiment 3. Steps and parameters: Skin irritation (rabbit model) Methods: The patch was applied on the back after hair removal for 48 hours, and the erythema and edema scores were observed (0-4 points).

[0041] Accelerated stability test Conditions: 40℃ / RH75% storage for 0, 1, 3, and 6 months, and testing the paste color, viscosity (Brookfield DV2T, 25℃) and active ingredient retention rate.

[0042] Expected results: Erythema score of all examples was ≤0.5 (1.5 for traditional plaster); The effective ingredient retention rate of Example 3 after 6 months is ≥90% (conventional plaster ≤60%).

[0043] Data Summary and Conclusions Although the present invention has been shown and described in conclusion: Embodiment 1: Achieve stable and long-lasting release, suitable for patients with chronic pain who need long-term application; Example 2: The antibacterial and anti-inflammatory properties are significantly better than those of traditional products, and are suitable for eczema and dermatitis with infection; Example 3: Rapid analgesia and microcirculation improvement capabilities are outstanding, meeting the needs of sports injury emergency treatment, and has the best high temperature stability. Experimental data confirms that each example has broken through the bottleneck of existing technologies in specific functional dimensions, and has clear clinical application scenarios.

[0044] Embodiments: For those of ordinary skill in the art, it will be understood that various changes, modifications, substitutions and variations may be made to these embodiments without departing from the principles and spirit of the invention, and the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A nanopatch, characterized in that: It includes 55%-60% nanoparticles and 40%-45% active ingredients of traditional Chinese medicine; the nanoparticles include: 25%-30% animal protein extract, 15%-20% mineral protein extract, and 50%-55% nanocarrier substrate.

2. A nano patch according to claim 1, characterized in that, The animal protein extract comprises: fish scale collagen.

3. A nano plaster according to claim 1, characterized in that, The mineral protein extract comprises: diatomaceous earth protein.

4. A nano patch according to claim 1, characterized in that, The active ingredients of the traditional Chinese medicine include: 12%-15% of auricularia auricula polysaccharide extract, 10%-15% of Panax notoginseng total saponin extract, 5%-10% of safflower flavonoids extract, 8%-12% of salvianolic acid extract, and 5%-10% of malic acid.

5. A method for preparing a nano-patch according to any one of claims 1 to 4, characterized in that: The following steps are involved: S1. Extraction of active ingredients from traditional Chinese medicine: Mix Auricularia auricula, Panax notoginseng, safflower, and Salvia miltiorrhiza in a mass ratio of 3:4:2:1, crush using a traditional Chinese medicine grinder for 6-8 minutes, add 10 times the amount of 70% ethanol after sieving, place in an ultrasonic extractor for extraction 3 times, each time for 30 minutes, combine the extracts and concentrate them by rotary evaporator until there is no alcohol taste, and finally dry them in a freeze dryer for 24-36 hours to obtain a composite extract powder; S2. Extraction of animal protein and mineral protein: Add decalcified fish scales to a constant temperature enzymatic hydrolysis tank, enzymatically hydrolyze with 5% pepsin at 45°C and 120rpm for 4 hours, centrifuge for 15 minutes, dialyze the supernatant and freeze-dry to obtain high-purity collagen powder; then enzymatically hydrolyze diatomaceous earth with neutral protease at 50°C and 150rpm for 6 hours, centrifuge and dry the filtrate with a spray dryer to obtain mineral protein powder with D50≤10μm S3. Preparation of porous nanoparticles: Mix animal protein and mineral protein powder and dissolve them in deionized water, add 1% ammonium bicarbonate foaming agent, freeze in liquid nitrogen and use vacuum freeze dryer to dry for 24 hours to form a skeleton. Put the skeleton into nano ball mill for 2 hours, pass through 200 mesh sieve to obtain 80-120nm porous nanoparticles; S4. Active ingredient loading: The composite extract and malic acid were dissolved in pH 5.5 citric acid buffer, added to porous nanoparticles, adsorbed in a constant temperature oscillator for 8 hours, centrifuged and dried in a vacuum drying oven for 6 hours, and the final water content was controlled to be ≤3%; S5. Paste molding and patch preparation: Mix sodium carboxymethyl cellulose, glycerol, azone and deionized water, homogenize with a high shear homogenizer for 10 minutes to form a gel, use a coater to coat the film, then cut into 5×7 cm patches and cover with release paper for packaging.

Images