Traditional Chinese medicine acupoint patch for treating peripheral facial paralysis and preparation method of traditional Chinese medicine acupoint patch

Abstract

The invention belongs to the field of biological medicine, and particularly discloses a traditional Chinese medicine acupoint patch for treating peripheral facial paralysis and a preparation method thereof. The traditional Chinese medicine composition is prepared from 14 raw materials including radix paeoniae alba, scorpio, pawpaw and the like according to a specific ratio, and is characterized in that firstly, supercritical CO2 is adopted for extracting fat-soluble components of snow lotus herb (40 DEG C / 25 MPa), enzymolysis is adopted for extracting radix paeoniae alba (albiflorin is larger than or equal to 90.3%), PLGA is adopted for wrapping hippocampus submicron powder (the encapsulation rate is larger than or equal to 85%), and active components are reserved to the maximum extent; (2) the acupoint patch is designed into a three-layer structure, middle-layer drug-loaded hydrogel is embedded into a soluble microneedle array (with the length of 800 microns) and pH response chitosan microspheres, and a composite penetration enhancer (azone + menthol) is added, so that temperature / pH dual intelligent drug release (the release speed is 2.8 times at the temperature of 35 DEG C, and the release speed is 3.1 times at the pH of 5.5) is realized; and (3) the nano pearl powder and the radix paeoniae alba extract are included by beta-cyclodextrin (the inclusion rate is greater than or equal to 91%), so that the stability is improved. Clinical verifications show that the total effective rate of combined western medicine treatment is 92%, which is significantly superior to that of a single western medicine group (76%, Plt; 0.05), and no new adverse reaction is generated.

Description

TECHNICAL FIELD

[0001] The application belongs to the field of biological medicine, and specifically discloses a traditional Chinese medicine acupoint patch for treating peripheral facial paralysis and a preparation method thereof. BACKGROUND

[0002] Peripheral facial paralysis (Bell's palsy) is an acute peripheral facial muscle paralysis caused by facial nerve non-specific inflammation, which belongs to the category of "mouth deviation" and "hanging line wind" in traditional Chinese medicine. At present, the treatment of western medicine mainly uses glucocorticoids and antiviral drugs, but long-term use can easily cause metabolic disorders. Traditional Chinese medicine often uses acupuncture combined with oral administration of traditional Chinese medicine, but oral administration has problems such as liver first-pass effect and gastrointestinal irritation. In the external treatment method, the traditional Chinese medicine acupoint patch becomes a research hotspot because it is directly applied to the disease and is easy to operate. However, the existing traditional Chinese medicine patches have three major defects: first, the transdermal efficiency of the compound medicine is low, the stratum corneum barrier makes it difficult for active ingredients (such as saponins and alkaloids) to penetrate into the deep layer of nerve and muscle, and the problem of transdermal penetration of macromolecules such as scorpion and cantharides has not been solved; second, the traditional patch lacks an intelligent release mechanism, and cannot respond to changes in facial temperature and pH fluctuations in the acupoint microenvironment, resulting in a drug utilization rate of less than 30%; third, the active ingredients of rare medicinal materials such as Saussurea involucrata and hippocampus are easily degraded in conventional extraction, such as the inactivation of Saussurea involucrata flavonoids when heated, which affects nerve repair efficacy.

[0003] The prior art attempts to improve the performance of the patch through a single technology, but none of them has achieved a systematic breakthrough, and has not been able to solve the three challenges of "transdermal depth-intelligent release-active retention" in a coordinated manner, and lacks hemodynamic adaptation design for facial paralysis acupoints (such as buccal car and ground warehouse), resulting in a long-term clinical effective rate of 40-60%. SUMMARY

[0004] In view of the above problems, the application provides a traditional Chinese medicine acupoint patch for treating peripheral facial paralysis and a preparation method thereof. By constructing an integrated technology system of "compound active retention-micro-needle deep delivery-intelligent response release", the following is adopted: supercritical extraction of fat-soluble components of Saussurea involucrata, PLGA nanoparticle encapsulation of hippocampus ultrafine powder, fusion of 800 μm soluble microneedles to penetrate the stratum corneum, 32 ℃ temperature-sensitive hydrogel to respond to body surface temperature, and pH 5.5 responsive chitosan microspheres to achieve three-level controlled release, combined with azaone-mint complex penetration enhancer and high inclusion rate technology of β-cyclodextrin (pearl powder / white peony glycoside ≥ 90%), which significantly improves the target enrichment efficiency of buccal car acupoint and ground warehouse acupoint, and breaks through the problems of limited efficacy and uncontrolled release of traditional patches.

[0005] The technical scheme of the application is as follows:

[0006] A traditional Chinese medicine composition for treating peripheral facial paralysis is prepared from the following raw materials by weight:

[0007] Radix Paeoniae Alba 40-50 parts, Scorpio 5-8 parts, Chaenomeles fruit 25-35 parts, Earthworm 10-15 parts, Salvia miltiorrhiza 25-35 parts, Chuanxiong 12-18 parts, Pueraria 25-35 parts, Bombyx Batryticatus 8-12 parts, Dendrobium officinale 8-10 parts, Angelica 8-12 parts, Spatholobus suberectus 12-18 parts, Saussurea involucrata extract 3-5 parts, Hippocampus super-fine powder 1-2 parts, nano pearl powder 2-3 parts.

[0008] The application further discloses a preparation method of the traditional Chinese medicine composition.

[0009] a) Saussurea involucrata extract preparation: supercritical CO2 extraction of Saussurea involucrata is carried out under the conditions of 40±2 ℃ and 25±1 MPa to obtain fat-soluble ingredients;

[0010] b) Radix Paeoniae Alba extract preparation: Radix Paeoniae Alba is extracted by using cellulase and pectinase complex enzyme under the conditions of pH 4.5±0.2 and 50±2 ℃, and after extraction, the Radix Paeoniae Alba is concentrated to a content of ≥92.7 %;

[0011] c) Hippocampus active component treatment: Hippocampus super-fine powder is coated with PLGA nanoparticles, and the encapsulation rate is ≥85 %;

[0012] d) Raw material treatment: Scorpio and Bombyx Batryticatus are separately ground into fine powder; and the remaining medicinal materials are crushed into fine powder with a particle size of ≤100 μm;

[0013] e) mixing the products obtained in a)-d) uniformly.

[0014] The application further discloses application of the traditional Chinese medicine composition in preparation of a peripheral facial paralysis acupoint patch.

[0015] The application further discloses a traditional Chinese medicine acupoint patch for treating peripheral facial paralysis, which comprises the traditional Chinese medicine composition and has the following structure:

[0016] Outer layer: medical polyurethane breathable film, thickness 50-100 μm;

[0017] Middle layer: drug-loaded hydrogel layer, containing a temperature-sensitive hydrogel matrix and the traditional Chinese medicine composition, and the LCST of the matrix is 32±1 ℃;

[0018] Inner layer: peelable silica gel anti-sticking layer.

[0019] Further, the traditional Chinese medicine acupoint patch,

[0020] The soluble microneedle array is embedded in the drug-loaded hydrogel of the middle layer, the microneedle length is 800±50 μm, and the drug loading amount is 0.8±0.1 mg / cm 2 ;

[0021] And the hydrogel further comprises a composite penetration enhancer: azone 1.0±0.2 wt %, menthol 3.0±0.5 wt %.

[0022] Further, the above-mentioned traditional Chinese medicine acupoint patch contains pH-responsive chitosan microspheres in the drug-loaded hydrogel, and the release rate of the microspheres in the skin pH 5.5 environment is increased by more than 2.5 times in the neutral environment.

[0023] The application further discloses a preparation method of the traditional Chinese medicine acupoint patch, which comprises the following steps:

[0024] S1, preparation of pH-responsive microspheres:

[0025] S11, dissolve chitosan in 1-2% acetic acid solution, and add white peony root extract, nano-level pearl powder and saussurea involucrata extract in the traditional Chinese medicine composition;

[0026] S12, continue to add 0.1-0.5 mol / L sodium citrate crosslinking agent, and solidify under the condition of pH 6-7 to obtain drug-loaded microspheres with a particle size of 50-200 μm;

[0027] S13, inject the drug-loaded microspheres into a microneedle mold, and solidify and form under the condition of vacuum at 25-30 DEG C to form a drug-loaded microneedle array;

[0028] S2, preparation of a drug-loaded hydrogel layer:

[0029] S21, add azone and menthol in the temperature-sensitive hydrogel matrix;

[0030] S22, further add other ingredients in the traditional Chinese medicine composition and mix uniformly;

[0031] S23, embed the drug-loaded microneedle array in step S13 into the drug-loaded hydrogel to prepare a drug-loaded hydrogel layer; the content of the traditional Chinese medicine composition accounts for 5-10 wt% of the mass of the final drug-loaded hydrogel layer, azone accounts for 1.0 ± 0.2 wt%, and menthol accounts for 3.0 ± 0.5 wt%;

[0032] S3, composite the drug-loaded hydrogel layer between a polyurethane breathable film and a silicone anti-adhesion layer, heat-press the edges, and the heat-press parameters are as follows: temperature 80 ± 5 DEG C, and pressure 0.1-0.2 MPa.

[0033] Further, in the above preparation method, the nano-level pearl powder and the white peony root extract are wrapped by beta-cyclodextrin, the wrapping ratio is 1:2-4, and the wrapping rate is greater than or equal to 90%.

[0034] The application discloses a facial paralysis treatment kit, comprising:

[0035] A traditional Chinese medicine acupoint patch for treating peripheral facial paralysis;

[0036] An acupoint positioning navigation patch, which is marked with the positions of buqicun points and dican points.

[0037] The application also discloses application of the saussurea involucrata extract in preparation of a traditional Chinese medicine acupoint patch for promoting facial nerve repair.

[0038] The extract is obtained by supercritical CO2 extraction, and the extraction conditions are 40+ / -2 DEG C and 25+ / -1 MPa.

[0039] Compared with the prior art, the application has the following beneficial effects:

[0040] The application discloses a traditional Chinese medicine acupoint patch for treating peripheral facial paralysis and a preparation method thereof, and makes progress in the field of traditional Chinese medicine external treatment for treating peripheral facial paralysis. In view of the problem that active ingredients of rare medicinal materials such as saussurea involucrata and hippocampus are easy to be inactivated, the application creatively adopts a triple protection strategy of'supercritical low-temperature extraction + enzymatic directional enrichment + nano wrapping', solves the problem of ingredient degradation caused by traditional heat extraction, and further creatively combines soluble microneedle penetration of the keratin barrier, temperature-sensitive hydrogel response to facial temperature intelligent dissolution, pH response microsphere targeted acupoint weak acid environment accelerated drug release and composite penetration enhancer, breaks through the technical bottleneck of 'difficult transdermal penetration and uncontrolled release' of traditional patches, realizes accurate enrichment of drugs in the facial nerve lesion area in theory, and proves the effect of the application through experiments. Further, the application reduces the irritation through the β-cyclodextrin inclusion technology, optimizes the structure of the patch by combining the anatomical characteristics of the buccal car and the dircang acupoint, and synchronously improves the patient compliance and the symptom relief speed, and clinical verification proves that the application can accelerate facial muscle function recovery and has better safety when combined with western medicine. Finally, the application constructs an integrated technology system of 'active preservation-delivery control-acupoint adaptation', and provides a solution with characteristics of traditional Chinese medicine syndrome differentiation and advantages of modern preparations for peripheral facial paralysis. BRIEF DESCRIPTION OF DRAWINGS

[0041] Figure 1 Comparison of nerve function score results in animal model efficacy verification test;

[0042] Figure 2 Comparison of NGF (pg / mL) content results in animal model efficacy verification test;

[0043] Figure 3 Comparison of axon regeneration density (%) results in animal model efficacy verification test. DETAILED DESCRIPTION

[0044] The technical solutions in the embodiments of the application will be described clearly and completely below. Apparently, the described embodiments are only part of the embodiments of the application, rather than all the embodiments. Based on the embodiments in the application, all other embodiments obtained by those skilled in the art without creative work fall within the protection scope of the application.

[0045] Table 1: raw material table

[0046] The application also discloses application of the saussurea involucrata extract in preparation of a traditional Chinese medicine acupoint patch for promoting facial nerve repair.

[0038] The extract is obtained by supercritical CO2 extraction, and the extraction conditions are 40+ / -2 DEG C and 25+ / -1 MPa.

[0039] Compared with the prior art, the application has the following beneficial effects:

[0040] The application discloses a traditional Chinese medicine acupoint patch for treating peripheral facial paralysis and a preparation method thereof, and makes progress in the field of traditional Chinese medicine external treatment for treating peripheral facial paralysis. In view of the problem that active ingredients of rare medicinal materials such as saussurea involucrata and hippocampus are easy to be inactivated, the application creatively adopts a triple protection strategy of'supercritical low-temperature extraction + enzymatic directional enrichment + nano wrapping', solves the problem of ingredient degradation caused by traditional heat extraction, and further creatively combines soluble microneedle penetration of the keratin barrier, temperature-sensitive hydrogel response to facial temperature intelligent dissolution, pH response microsphere targeted acupoint weak acid environment accelerated drug release and composite penetration enhancer, breaks through the technical bottleneck of 'difficult transdermal penetration and uncontrolled release' of traditional patches, realizes accurate enrichment of drugs in the facial nerve lesion area in theory, and proves the effect of the application through experiments. Further, the application reduces the irritation through the β-cyclodextrin inclusion technology, optimizes the structure of the patch by combining the anatomical characteristics of the buccal car and the dircang acupoint, and synchronously improves the patient compliance and the symptom relief speed, and clinical verification proves that the application can accelerate facial muscle function recovery and has better safety when combined with western medicine. Finally, the application constructs an integrated technology system of 'active preservation-delivery control-acupoint adaptation', and provides a solution with characteristics of traditional Chinese medicine syndrome differentiation and advantages of modern preparations for peripheral facial paralysis. BRIEF DESCRIPTION OF DRAWINGS

[0041] Figure 1 Comparison of nerve function score results in animal model efficacy verification test;

[0042] Figure 2 Comparison of NGF (pg / mL) content results in animal model efficacy verification test;

[0043] Figure 3 Comparison of axon regeneration density (%) results in animal model efficacy verification test. DETAILED DESCRIPTION

[0044] The technical solutions in the embodiments of the application will be described clearly and completely below. Apparently, the described embodiments are only part of the embodiments of the application, rather than all the embodiments. Based on the embodiments in the application, all other embodiments obtained by those skilled in the art without creative work fall within the protection scope of the application.

[0045] Table 1: raw material table

[0046]

[0047]

[0048] Note: The rest of the medicinal materials (papaya, earthworm, etc.) meet the standards of Chinese Pharmacopoeia 2020 edition, and the sources are GMP certified enterprises such as Tong Ren Tang Pharmaceutical Co., Ltd. and China Resources Sanjiu.

[0049] Table 2: Instrument list

[0050]

[0051] Example 1

[0052] Raw material ratio (weight parts): Radix Paeoniae Alba 40, Scorpio 5, Papaya 25, Earthworm 10, Salvia Miltiorrhiza 25, Chuanxiong 12, Pueraria 25, Bombyx Batryticatus 8, Dendrobium officinale 8, Angelica 8, Spatholobus suberectus 12, Saussurea involucrata extract 3, Hippocampus super-fine powder 1, nano pearl powder 2.

[0053] Preparation method:

[0054] 1. Saussurea involucrata extract:

[0055] Saussurea involucrata dry powder (passed through an 80-mesh sieve) was mixed with CO2 at a mass ratio of 1:15 into the extraction kettle;

[0056] Supercritical CO2 extraction: temperature 40±0.5℃, pressure 25±0.2MPa, CO2 flow rate 25L / h, extraction time 2.5h;

[0057] The fat-soluble ingredients were collected in the separation kettle, with a yield of 8.7%.

[0058] 2. Radix Paeoniae Alba extract:

[0059] Radix Paeoniae Alba slices were infiltrated with 10 times the amount of pH 4.5 acetic acid-sodium acetate buffer;

[0060] Compound enzyme (cellulase 1200U / g raw material + pectinase 800U / g raw material) was added, and enzyme hydrolysis was carried out at 50±0.5℃ for 3h;

[0061] Boiling water was used to inactivate the enzyme for 10min, then filtered, and the filtrate was concentrated under reduced pressure to a relative density of 1.25 (60℃);

[0062] Spray drying (inlet temperature 160℃, outlet temperature 85℃) was carried out to obtain Radix Paeoniae Alba extract (albiflorin content 90.3%).

[0063] 3. Hippocampus active component:

[0064] Hippocampus super-fine powder (particle size ≤10μm) was dispersed in dichloromethane at a mass ratio of 1:5 with PLGA (50:50, MW 15kDa);

[0065] Dropping into 1% PVA aqueous solution, homogenizing emulsification (10000 rpm, 10 min);

[0066] Volatile organic solvent, centrifugal collection of PLGA nanoparticles (encapsulation efficiency 86.2%, particle size 210 ± 30 nm).

[0067] 4, raw material processing:

[0068] Scorpion, silkworm powder alone (particle size ≤ 50 μm);

[0069] Papaya, earthworm and other remaining medicinal materials are mixed and crushed to a particle size of ≤ 100 μm.

[0070] Mix all components to obtain a traditional Chinese medicine composition.

[0071] Example 2-3

[0072] The raw material ratio is shown in Table 3, and the preparation method is the same as in Example 1.

[0073] Table 3: Raw material ratio (parts by weight) of Example 2-3,

[0074] Ingredients Example 2 Example 3 Radix Paeoniae Alba 45 50 Saussurea Involucrata Extract 4 5 Hippocampus Superfine Powder 1.5 2

[0075] The same as the scope of claim 1.

[0076] Example 4

[0077] The same as Example 2, with the addition of the following steps:

[0078] β-cyclodextrin inclusion: Nanometer pearl powder and white peony extract were mixed at a ratio of 1:3, β-cyclodextrin saturated aqueous solution (60°C) was added, magnetic stirring for 2 h, 4°C standing for 12 h, filtration and drying (inclusion rate 91.5%).

[0079] Comparative Example 1

[0080] The same as Example 2, but omitting the Saussurea extract.

[0081] Comparative Example 2

[0082] The same as Example 2, but the hippocampus ultrafine powder is not PLGA coated (directly using ultrafine powder).

[0083] Comparative Example 3

[0084] The same as Example 2, but the Saussurea extract is replaced by ethanol hot extraction: Saussurea powder is extracted with 70% ethanol by reflux (solid-liquid ratio 1:10, 80°C, 2 h), and after recovering the ethanol, freeze-drying.

[0085] Comparative Example 4

[0086] The same as Example 2, but the white peony extract is replaced by water extraction and alcohol precipitation:

[0087] White peony root decoction 2 times (1:8 water, 1 h / time), combined filtrate, concentrated to 1:1, ethanol to contain 70%, 70% alcohol, take the precipitate dry (baeocin content 78.4%).

[0088] Comparative example 5

[0089] The same as example 2, but replace Dendrobium candidum with equal amount of Astragalus (change the composition of the monarch, minister, and assistant).

[0090] Application example 1

[0091] S1. Preparation of pH-responsive microspheres:

[0092] S11: 2.5 g of chitosan (degree of deacetylation ≥95%) was dissolved in 150 ml of 1.5% acetic acid solution;

[0093] S12: Add 1.2 g of white peony root extract, 0.4 g of nano pearl powder, and 0.15 g of saussurea involucrata extract obtained in example 1, and homogenize and disperse (5000 rpm, 10 min);

[0094] S13: Add 0.3 mol / L sodium citrate solution dropwise (flow rate 2 ml / min), control temperature 25℃, magnetic stirring (300 rpm) until pH 6.5, solidify for 1 h;

[0095] S14: Centrifugal collection of microspheres (particle size 120±30 μm), distilled water washing 3 times, freeze-drying.

[0096] S2. Formation of drug-loaded microneedle array:

[0097] Disperse the microspheres in 20% PVP K90 aqueous solution and inject into a polydimethylsiloxane (PDMS) microneedle mold (needle height 800 μm, base diameter 200 μm);

[0098] Vacuum drying at 25℃ (-0.09 MPa, 2 h), demolding to obtain a drug-loaded microneedle array.

[0099] S3. Preparation of drug-loaded hydrogel layer:

[0100] Take 10 g of temperature-sensitive hydrogel matrix (PNIPAM, LCST 32℃), add 0.10 g of azone and 0.30 g of menthol;

[0101] Add the remaining traditional Chinese medicine composition (scorpion powder, silkworm chrysalis powder, etc.) 0.85 g, and homogenize and mix;

[0102] Embed the microneedle array obtained in S2 (drug loading 0.82 mg / cm 2 ), and the total content of traditional Chinese medicine accounts for 8.5 wt% of the hydrogel layer.

[0103] S4. Composite packaging:

[0104] Drug-loaded hydrogel layer was coated on polyurethane breathable film (80 μm);

[0105] Covered with silicone release layer (thickness 100 μm);

[0106] Hot-pressing edge (80℃±1℃, 0.15 MPa, 10 s holding pressure).

[0107] Application Example 2-3

[0108] Application Example 2: The composition of Example 4 (containing β-cyclodextrin inclusion compound) was used, and the rest was the same as in Application Example 1.

[0109] Application Example 3: The composition of Comparative Example 2 (without hippocampus wrapping) was used, and the rest was the same as in Application Example 1.

[0110] Test Example 1

[0111] Purpose of active ingredient retention rate test: to verify the protective effect of supercritical CO2 extraction (Saussurea involucrata) and enzymatic process (Radix Paeoniae Alba) on heat-sensitive ingredients.

[0112] Method:

[0113] HPLC was used to detect the content of key ingredients:

[0114] Saussurea involucrata extract: syringin (representative of flavonoids)

[0115] Radix Paeoniae Alba extract: paeonoside

[0116] Hippocampus PLGA nanoparticles: polypeptide content (Bradford method)

[0117] Comparison of ingredient retention rate (%) of different extraction processes = (actual content / theoretical maximum content) x 100%.

[0118] The results are shown in Table 4

[0119] Table 4 Active ingredient retention rate test

[0120] Group Retention Rate of Syringin Content of Albasaponin Retention Rate of Hippocampus Polypeptide Example 2 95.2±1.8％ 90.8±0.9％ 92.5±2.1％ Comparative Example 3 63.7±3.5％ - - Comparative Example 4 - 78.4±1.2％ - Comparative Example 2 - - 71.3±4.0％

[0121] From Table 4, the following conclusions can be drawn:

[0122] ① Supercritical CO2 (Example 2) significantly improved the retention rate of Saussurea involucrata flavonoids compared to ethanol hot extraction (Comparative Example 3) (p<0.01);

[0123] ② Enzymatic method (Example 2) increased the content of paeonoside by 12.4% compared to water extraction and alcohol precipitation (Comparative Example 4);

[0124] iii. PLGA encapsulation (Example 2) effectively protects Hippocampus peptide activity (retention rate > 90%).

[0125] Test Example 2

[0126] Transdermal efficiency and smart release evaluation

[0127] Objective: To verify the synergistic transdermal effect and temperature / pH responsive properties of microneedles + co-penetration enhancer + pH-responsive microspheres.

[0128] Methods:

[0129] In vitro transdermal experiment (Franz diffusion cell):

[0130] Skin model: ex vivo pig ear skin (thickness 0.8 ± 0.1 mm)

[0131] Test components: tanshinone IIA (representative liposoluble component), paeoniflorin (representative water-soluble component)

[0132] Grouping: Application Example 1 (complete design) vs. Application Example 3 (without PLGA encapsulation) vs. traditional patch (without microneedles / penetration enhancer)

[0133] Smart release test:

[0134] Temperature response: compare the cumulative release rate of paeoniflorin at 25°C (room temperature) vs. 35°C (facial temperature);

[0135] pH response: compare the release rate of tanshinone IIA in pH 7.4 (neutral) vs. pH 5.5 (skin) environments.

[0136] Results Table 5 and Table 6:

[0137] Table 5: 24h cumulative transdermal amount (μg / cm 2 )

[0138] Group Tanshinone IIA Paeoniflorin Application Example 1 38.7±2.1 52.6±3.8 Application Example 3 19.2±1.6 33.4±2.7 Traditional Patch 5.3±0.9 12.1±1.4

[0139] Table 6 Smart release performance

[0140] Conditions Application Example 1 (Paeoniflorin) Application Example 1 (Tanshinone IIA) Release Increase at 35℃ vs 25℃ 2.8 times - Release Rate Ratio at pH 5.5 vs 7.4 - 3.1 times

[0141] From Table 5 and 6, the following conclusions can be drawn:

[0142] i. Microneedles + penetration enhancer (Application Example 1) make the transdermal amount 7.3 times that of the traditional patch (tanshinone);

[0143] ii. Temperature-sensitive hydrogel triggers drug burst release at 35°C (facial temperature) (LCST 32°C);

[0144] iii. pH-responsive microspheres accelerate drug release in the weakly acidic environment of the skin (release rate increased by 3.1 times).

[0145] Test Example 3

[0146] Animal model efficacy verification

[0147] Objective: To evaluate the promoting effect of acupoint paste on facial nerve function recovery.

[0148] Methods:

[0149] Model establishment: SD rat facial nerve crush injury model (n = 8 / group);

[0150] Group intervention:

[0151] Blank group: sham operation

[0152] Model group: injury + normal saline

[0153] Positive group: injury + prednisone tablets (3 mg / kg / d)

[0154] Experimental group: injury + application example 1 acupoint paste (apply cheek car / ground warehouse acupoint, 8h daily)

[0155] Control group: injury + comparative example 1 acupoint paste (no saussurea extract)

[0156] Evaluation index:

[0157] Facial nerve function score (facial nerve function score uses House-Brackmann grading standard: I grade (1 point) normal, VI grade (6 points) complete paralysis, the higher the score, the worse the function);

[0158] Histopathology: HE staining to observe facial nerve axon regeneration;

[0159] ELISA detection: serum NGF (nerve growth factor) level.

[0160] Results (after 14 days of treatment) are shown in Table 7 and Figures 1-3 .

[0161] Table 7 Animal model efficacy verification

[0162]

[0163] From Table 7, we can conclude:

[0164] ① Application example 1 significantly improved nerve function (score was significantly reduced compared with the model group, p < 0.01 vs positive group);

[0165] ② NGF level close to normal group (blank group), confirming the nerve repair effect of saussurea extract;

[0166] Example 3 (without Saussurea involucrata) significantly reduced efficacy, proving the necessity of the integrity of the formula.

[0167] Test Example 4

[0168] Formulation stability and irritation

[0169] Objective: To verify the improvement of ingredient stability and formulation safety by β-cyclodextrin inclusion technology.

[0170] Grouping: Example 1 (without inclusion) vs. Example 2 (with β-cyclodextrin inclusion)

[0171] Results are shown in Table 8

[0172] Table 8 Formulation stability and irritation test

[0173] Test Items Application Example 1 Application Example 2 Retention Rate of Paeoniflorin after 30 Days of Storage at 40℃ 82.3±2.1％ 95.6±1.5％ Skin Irritation Index (PII) 0.48 (mild irritation) 0.21 (no irritation) Dissolution Time of Microneedle (s) 35.2±3.1 28.7±2.4

[0174] From the above data, it can be concluded that β-cyclodextrin inclusion (Example 2) significantly improves drug stability, reduces irritation, and accelerates microneedle dissolution (p<0.05).

[0175] Test Example 5

[0176] Clinical efficacy control test (200 RCT retrospective analysis)

[0177] Objective: To verify the clinical advantages of acupoint patch combined with Western medicine compared with Western medicine alone.

[0178] Grouping and methods:

[0179] Test group (n=100): prednisone (oral, 60mg / d, halved after 3 days) + methocel (0.5mg tid) + acupuncture + Example 1 acupoint patch (applied to the cheek car / ground warehouse acupoint, 8h daily)

[0180] Control group (n=100): same Western medicine + acupuncture regimen (without acupoint patch)

[0181] Course of treatment: 4 weeks

[0182] Evaluation criteria:

[0183] Total effective rate: "Peripheral facial nerve palsy efficacy determination criteria"

[0184] Cure (47-50 points) | Markedly effective (↑>15 points) | Effective (↑>10 points) | Ineffective (↑≤10 points)

[0185] Facial nerve function:

[0186] House-Brackmann (H-B) score (1-6 points, ≤3 points good)

[0187] Sunnybrook (SFGS) score (0-100,↑=improvement

[0188] Results are shown in Tables 9-11:

[0189] Table 9: Total effective rate comparison

[0190] Group Cured Markedly Effective Effective Ineffective Total Effective Rate Test Group 38 42 12 8 92％ Control Group 22 34 20 24 76％

[0191] Statistics: χ 2 =9.32, P<0.05

[0192] Table 10: Changes in facial nerve function score

[0193]

[0194]

[0195] Table 11: Incidence of adverse reactions

[0196] Group Dizziness Skin Rash Lassitude Total Incidence P Value Test Group 2 2 2 6％ 0.774 Control Group 2 2 3 7％ -

[0197] From Tables 9-11, the following conclusions can be drawn:

[0198] ①The total effective rate of the test group was significantly better than that of the control group (92% vs 76%, P<0.05);

[0199] ②The improvement in H-B and SFGS scores was significantly higher than that of the control group (P=0.001);

[0200] ③Acupoint sticking did not increase adverse reactions (P>0.05).

[0201] Typical cases:

[0202] Zhang XX, male, 18 years old: left facial paralysis for 1 day, manifested as disappearance of left forehead lines, incomplete eye closure (slit 4 mm), and air leakage from the cheek. Apply Example 1 acupoint stickers to the affected side of the cheek car and Dicang points. After 24 hours: eye closure function was significantly restored (slit reduced to 1 mm), and frowning movements could be completed.

[0203] Sun XX, female, 25 years old: right facial paralysis for 3 days, accompanied by severe pain behind the ear (VAS score 7). After 5 hours of application of Example 1 acupoint stickers, the pain behind the ear decreased to VAS 3; after 3 days of treatment: the deviation of the mouth corner improved, and the salivation during eating decreased by 50%.

[0204] Liu XX, male, 35 years old: facial paralysis worsened after hormone treatment, left nasolabial sulcus became shallow, and food retention occurred during chewing. After 7 days of combined application of Example 1 stickers: facial muscle numbness decreased, there was no air leakage from the cheek, and dynamic facial expression recovered symmetrically.

[0205] Dong XX, female, 50 years old: facial paralysis for 4 days with dry mouth and tongue (saliva secretion test < 0.5 ml / min). Application of Example 1 + Western medicine treatment for 2 weeks: House-Brackmann score improved from 4 to 2, and saliva secretion recovered to 1.2 ml / min.

[0206] Wang XX, female, 55 years old: facial paralysis for 1 day with insomnia (PSQI score 15). Application of Example 1 for 2 weeks: 90% recovery of facial muscle motor function, and PSQI score reduced to 6 (sleep latency shortened to 20 minutes).

[0207] Test Example 6

[0208] Formulation optimization clinical verification (150 cases grouped)

[0209] Objective: To compare the clinical effectiveness of different formulation processes (based on Application Examples 1-3)

[0210] Grouping:

[0211] Group A (n = 50): Application Example 1 acupoint patch (complete design)

[0212] Group B (n = 50): Application Example 2 acupoint patch (containing β-cyclodextrin inclusion)

[0213] Group C (n = 50): Application Example 3 acupoint patch (Hai Ma not wrapped with PLGA)

[0214] Treatment regimen: Same as Test Example 5 test group (Western medicine + acupuncture + acupoint patch)

[0215] Results are shown in Table 12

[0216] Table 12 Formulation optimization clinical verification

[0217] Group Total Effective Rate H-B Score after Treatment (mean ± standard deviation) SFGS Score after Treatment (mean ± standard deviation) Group A 90％ 1.62±0.25 90.35±7.15 Group B 94％ 1.48±0.19 93.21±6.84 Group C 82％ 1.95±0.31 85.47±8.02

[0218] From Table 12, the following conclusions can be drawn:

[0219] ① Group B (β-cyclodextrin inclusion) has the best efficacy (efficiency 94%), confirming that the inclusion technology improves the stability and bioavailability of the components;

[0220] ② Group C (Hai Ma not wrapped with PLGA) has significantly reduced efficacy, verifying the necessity of PLGA wrapping for the protection of nerve repair components.

[0221] From the above test examples, it can be seen that the present application has the following progressiveness:

[0222] The present application, through the synergistic innovation of compound optimization and delivery system, breaks through the three technical bottlenecks of traditional Chinese medicine patches:

[0223] 1. Active ingredient high-efficiency reservation and utilization: ①Supercritical CO2 extraction of Saussurea involucrata (conditions 40℃ / 25MPa) makes the reservation rate of syringin reach 95.2%, which is 49.3% higher than that of the ethanol hot extraction method (63.7%); ②White peony root enzymatic extraction (cellulase + pectinase, pH 4.5 / 50℃) ensures that the content of paeonoside is ≥90.3%, which is 15.2% higher than that of the water extraction and alcohol precipitation method (78.4%); ③PLGA nanoparticle-encapsulated hippocampus ultrafine powder (encapsulation rate 86.2%) makes the reservation rate of polypeptide reach 92.5%, solving the problem of animal medicine activity degradation (test example 1).

[0224] 2. Multi-stage intelligent transdermal delivery breaks through the barrier: ①Soluble microneedles (800μm) penetrate the stratum corneum, combined with azone-mint penetration enhancer, making the transdermal amount of tanshinone IIA reach 38.7μg / cm 2 , which is 7.3 times that of traditional patches (test example 2); ②Temperature-sensitive hydrogel (LCST 32℃) triggers burst drug release at facial temperature, and the release amount of paeonol increases by 2.8 times at 35℃; ③pH-responsive chitosan microspheres accelerate drug release in the weakly acidic environment of the skin (pH 5.5), and the release rate of tanshinone IIA is 3.1 times that in a neutral environment (test example 2).

[0225] 3. Clinical efficacy and safety double verification: ①A retrospective analysis of 200 cases showed that the total effective rate of combined treatment with western medicine was 92%, which was significantly better than that of the single western medicine group (76%, P<0.05), the improvement amplitude of House-Brackmann score (↓1.90 vs ↓1.32) and Sunnybrook score (↑29.48 vs ↑19.74) were statistically different (P=0.001), and the adverse reaction rate was only 6% (no difference with the western medicine group 7%, P>0.05) (test example 5); ②β-cyclodextrin inclusion technology (application example 2) further improves the clinical effective rate to 94%, while the hippocampus preparation without packaging (application example 3) reduces to 82% (test example 6); ③Typical cases show that the pain behind the ear is relieved 5 hours after the application, and the sleep is improved after 2 weeks, highlighting the advantages of quick-acting and continuous repair.

[0226] 4. The stability of the preparation is significantly improved: β-cyclodextrin inclusion makes the reservation rate of paeonol reach 95.6% (82.3% for the non-inclusion group) at 40℃ for 30 days, and the skin irritation index is reduced to 0.21 (non-irritating) (test example 4), providing protection for industrialization.

[0227] The above is only a limited number of preferred embodiments of the present application, which is described in more detail and in detail, but it cannot be interpreted as a limitation on the scope of the patent of the present application. It should be noted that for ordinary skilled persons in the art, without departing from the concept of the present application, a number of modifications and improvements can be made, which are within the scope of protection of the present application.

Claims

1. A Chinese medicine composition for treating peripheral facial paralysis, characterized in that: Made from the following raw materials in parts by weight: 40-50 parts of white peony root, 5-8 parts of scorpion, 25-35 parts of papaya, 10-15 parts of earthworm, 25-35 parts of salvia miltiorrhiza, 12-18 parts of chuanxiong, 25-35 parts of kudzu root, 8-12 parts of bombyx batryticatus, 8-10 parts of dendrobium officinale, 8-12 parts of angelica, 12-18 parts of millettia reticulata, 3-5 parts of snow lotus extract, 1-2 parts of hippocampus ultrafine powder, and 2-3 parts of nano-grade pearl powder.

2. The method for preparing the Chinese medicine composition according to claim 1, wherein: The following steps are involved: a) Preparation of Snow Lotus Extract: Snow Lotus was extracted using supercritical CO2 at 40±2°C and 25±1 MPa to obtain fat-soluble components; b) Preparation of white peony root extract: White peony root was extracted with a combined enzymatic hydrolysis of cellulase and pectinase at pH 4.5±0.2 and 50±2°C, and concentrated to a paeony glycoside content of ≥92.7%. c) Treatment of hippocampus active components: ultrafine hippocampus powder was encapsulated with PLGA nanoparticles, with an encapsulation efficiency of ≥85%; d) Raw material processing: Grind the scorpion and silkworm into fine powder; crush the remaining medicinal materials into fine powder with a particle size of ≤100 μm; e) Mixing the products obtained in steps a) to d) uniformly.

3. Use of the Chinese medicine composition as claimed in claim 1 in preparing acupoint plasters for peripheral facial paralysis.

4. A Chinese medicine acupoint patch for treating peripheral facial paralysis, characterized in that: The Chinese medicine composition according to claim 1 has the following structure: Outer layer: medical polyurethane breathable film, thickness 50-100μm; Middle layer: drug-loaded hydrogel layer, containing a temperature-sensitive hydrogel matrix and a traditional Chinese medicine composition, wherein the LCST of the matrix is ​​32±1°C; Inner layer: Peelable silicone anti-stick layer.

5. The Chinese medicine acupuncture patch according to claim 4, characterized in that: The drug-loaded hydrogel in the middle layer is embedded with a soluble microneedle array, with a microneedle length of 800±50μm and a drug loading capacity of 0.8±0.1mg / cm 2 ; The hydrogel further comprises a composite penetration enhancer: 1.0±0.2 wt % of azone and 3.0±0.5 wt % of menthol.

6. The Chinese medicine acupuncture patch according to claim 5, characterized in that: The drug-loaded hydrogel also contains pH-responsive chitosan microspheres, the release rate of which in a skin pH environment of 5.5 is increased to more than 2.5 times that in a neutral environment.

7. The method for preparing the Chinese medicine acupuncture patch according to claim 6, wherein: The following steps are involved: S1. Preparation of pH-responsive microspheres: S11, dissolving chitosan in 1-2% acetic acid solution, and adding the white peony root extract, nano-sized pearl powder and snow lotus extract in the traditional Chinese medicine composition; S12, continue to dropwise add 0.1-0.5 mol / L sodium citrate crosslinker, solidify at pH 6-7, and obtain drug-loaded microspheres with a particle size of 50-200 μm; S13, injecting the drug-loaded microspheres into the microneedle mold, and curing and molding them under vacuum conditions at 25-30° C. to form a drug-loaded microneedle array; S2. Preparation of drug-loaded hydrogel layer: S21, adding azone and menthol to the temperature-sensitive hydrogel matrix; S22, further adding other ingredients in the Chinese medicine composition and mixing evenly; S23, embedding the drug-loaded microneedle array of step S13 into the drug-loaded hydrogel to prepare a drug-loaded hydrogel layer; the content of the traditional Chinese medicine composition accounts for 5-10wt% of the mass of the final drug-loaded hydrogel layer, azone 1.0±0.2wt%, and menthol 3.0±0.5wt%; S3. Compound the drug-loaded hydrogel layer between the polyurethane breathable membrane and the silicone anti-sticking layer, and seal the edges by hot pressing. The hot pressing parameters are: temperature 80±5°C, pressure 0.1-0.2MPa.

8. The preparation method according to claim 7, characterized in that: The nano-scale pearl powder and the white peony root extract are included by beta-cyclodextrin, the inclusion ratio is 1:2-4, and the inclusion rate is ≥90%.

9. A facial paralysis treatment kit comprising: The facial paralysis acupuncture patch according to claims 4-6; Acupoint positioning navigation patch with the locations of Cheek-Riding Point and Dicang Point marked on it.

10. The use of snow lotus extract in the preparation of a traditional Chinese medicine acupoint patch for promoting facial nerve repair, characterized in that: The extract is obtained by supercritical CO2 extraction under the following conditions: 40±2°C and 25±1MPa.

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