Traditional Chinese medicine prescription for traditional Chinese medicine acupuncture and preparation method thereof

Abstract

The invention discloses a traditional Chinese medicine prescription for traditional Chinese medicine acupuncture and a preparation method thereof and belongs to the technical field of traditional Chinese medicine. According to the prescription, astragalus membranaceus and salvia miltiorrhiza are used as a core drug pair, Chinese angelica, ligusticum wallichii and other blood circulation promoting and collateral dredging drugs are matched, safflower (hydroxysafflor yellow A is larger than or equal to 2%) and enzymolysis earthworm (smaller than or equal to 5kDa peptide fragment is larger than or equal to 60%) are used as auxiliary materials to enhance transdermal absorption, and liquorice is used for blending all the drugs. The preparation method comprises the steps of superfine grinding (less than or equal to 50 microns), ultrasonic-alcohol-water combined extraction, gradient vacuum concentration and macroporous resin purification, and can be prepared into dosage forms such as acupoint application gel, enteric-coated tablets and the like. The quality is controlled by combining HPLC fingerprint spectrum with multi-index component determination (astragaloside is greater than or equal to 0.8 mg / g, and tanshinone IIA is greater than or equal to 0.5 mg / g). Experiments show that the dissolution rate of the tanshinone IIA is increased by 49.2% through superfine grinding, the 24-hour transdermal permeation amount of the gel is 3.12 times that of a traditional decoction, the total effective rate of acupuncture and moxibustion combined medication reaches 95.0%, the analgesic and anti-inflammatory effects are remarkably enhanced, and the process is advanced and suitable for industrial production.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine, and specifically to a traditional Chinese medicine formula for traditional Chinese medicine acupuncture and a preparation method thereof comprising a superfine pulverization-ultrasonic assisted extraction process, especially applicable to the synergistic use of acupuncture in the treatment of pain syndromes caused by qi stagnation and blood stasis. Background Art

[0002] As a characteristic therapy of traditional Chinese medicine, acupuncture regulates the flow of qi and blood by stimulating the meridians and acupoints, and has irreplaceable advantages in the treatment of chronic pain (such as lumbar muscle strain, knee osteoarthritis), sequelae of stroke and other diseases. Clinical practice shows that acupuncture combined with oral or external use of traditional Chinese medicine can significantly improve the curative effect, but there are many technical bottlenecks in the existing acupuncture-assisted traditional Chinese medicine in terms of formula, process, preparation and quality control.

[0003] Traditional Chinese medicines used in traditional acupuncture mostly follow empirical formulas, with vague ratios of core medicine pairs and lack of collaborative design of active ingredients. Taking the classic qi-tonifying and blood-activating medicine pair of Astragalus membranaceus - Salvia miltiorrhiza as an example, the existing technology does not clarify the optimal ratio of the two, resulting in a dissolution rate of astragaloside IV and tanshinone IIA of only 45%-50%, making it difficult to fully exert the synergistic effect of "tonifying qi and dredging collaterals - promoting blood circulation to relieve pain". At the same time, there is a lack of quality control for key components of transdermal absorption. The content fluctuation of hydroxysafflor yellow A in safflower reaches 1%-5%, and the proportion of active peptide segments (≤5 kDa) in earthworm after simple pulverization is less than 50%, seriously restricting the transdermal efficiency of the drug.

[0004] At the level of the preparation process, the dissolution rate of fat-soluble components (such as tanshinone IIA) by the traditional water decoction method is only 48.6%, and the loss rate of the heat-sensitive component tetramethylpyrazine due to high-temperature decoction exceeds 30%; conventional pulverization through an 80-mesh sieve (particle size ≤180 μm) results in a cell wall breaking rate of less than 60% of the medicinal materials, and the dissolution rate of water-soluble components such as paeoniflorin is 25%-30% lower than that of the superfine pulverization process; the purification process is extensive and not refined according to the multi-target action characteristics of acupuncture. The content of active ingredients in the existing ointments decreases by more than 15% after 3 months of storage.

[0005] Another prominent problem is the single form of the preparation and poor adaptability. Decoctions need to be decocted immediately before use, with low patient compliance and unable to meet the transdermal drug delivery requirements of acupoint application; traditional ointments use matrices such as vaseline, with poor adhesiveness and uncontrollable release rate, and the cumulative penetration amount in 24 h is only 1 / 3 of that of the new gel; oral preparations lack dosage form optimization and do not adopt enteric coating technology, resulting in the degradation of active ingredients in gastric acid and a reduction in bioavailability of more than 20%.

[0006] In terms of quality control, existing technologies mostly use a single component (such as astragaloside IV) as the content determination index, ignoring the quality control of transdermal-related components such as hydroxysafflor yellow A and earthworm active peptide segments, and unable to ensure the consistency of curative effects between batches. At the same time, the lack of control over preparation characteristic parameters such as the viscosity of gels and the disintegration time limit of tablets directly affects the clinical use effect.

[0007] In response to the above problems, there is an urgent need to develop a traditional Chinese medicine prescription and its preparation method that has precise raw material processing, advanced preparation technology, and is suitable for combined acupuncture and moxibustion treatment. Summary of the Invention

[0008] The purpose of the present invention is to provide a traditional Chinese medicine prescription for acupuncture and a preparation method thereof, and to achieve technological breakthroughs through precise formula design, innovative preparation process, multi-formulation preparation development and strict quality control: clarifying the optimal ratio of the Astragalus membranaceus-Danshen medicinal pair (1:0.75-1.25) and introducing quantitative control of the active ingredients, using ultrafine grinding (≤50μm), ultrasound-assisted extraction, gradient decompression concentration and other technologies to improve the utilization efficiency of the ingredients, developing dosage forms such as acupoint patch gel and enteric-coated tablets suitable for acupuncture treatment, and establishing a quality standard combining HPLC fingerprint analysis with multi-index determination to ensure that the material basis of the efficacy is stable and controllable.

[0009] In order to achieve the above object, the present invention is implemented through the following technical solution: a traditional Chinese medicine prescription for acupuncture and moxibustion, comprising the following raw materials in parts by weight:

[0010] The core herbal pair for invigorating Qi: 20-30 parts of Astragalus and 15-25 parts of Salvia miltiorrhiza, with a weight ratio of 1:0.75-1.25;

[0011] Blood circulation and collateral dredging group: 15-25 parts of Chinese Angelica sinensis, 10-20 parts of Chuanxiong, 10-20 parts of Red Peony Root, 10-15 parts of Achyranthes bidentata;

[0012] Transdermal enhancement group: 10-15 parts of safflower, with a hydroxysafflower yellow A content of ≥2%; 10-15 parts of earthworm, with a peptide fraction of ≤5kDa after enzymatic hydrolysis accounting for ≥60%;

[0013] Blending adjuvants: 10-15 parts of peach kernel and 5-10 parts of licorice.

[0014] As a further improvement to the technical solution of the present invention, the enzymatic hydrolysis treatment of the earthworm is as follows: after freeze-drying at -50°C, dissolving in a buffer solution with a pH of 7.5-8.5 at a mass-to-volume ratio of 1:8 to 1:12, adding trypsin accounting for 1%-2% of the mass of the earthworm, enzymatic hydrolysis at 45-55°C for 3-5 hours, inactivating the enzyme, centrifuging, and taking the supernatant for freeze-drying.

[0015] As a further improvement to the technical solution of the present invention, the weight ratio of the Astragalus membranaceus-Danshen miltiorrhiza medicinal pair is 25:20.

[0016] As a further improvement of the technical solution of the present invention, the acupuncture synergistic preparation of the traditional Chinese medicine prescription is an acupoint application gel, which contains the following excipients in the following mass percentages:

[0017] Matrix: Carbomer 940 accounts for 1.0% - 2.0%, glycerol accounts for 4% - 6%, and triethanolamine accounts for 0.3% - 0.7%.

[0018] Penetration enhancer: Azone accounts for 0.3% - 0.7%.

[0019] Extract powder: Accounts for 15% - 25% based on crude drugs, and the rest is deionized water.

[0020] Furthermore, as an improvement to the technical solution of the present invention, the acupoint application gel is controlled for its preparation viscosity by rheological detection: at 25°C, the dynamic viscosity is 1000 - 1500 mPa·s, and the yield stress ≤ 5 Pa.

[0021] Furthermore, as an improvement to the technical solution of the present invention, a preparation method of a traditional Chinese medicine formula for traditional Chinese medicine acupuncture includes the following steps:

[0022] Ultra-fine pulverization pretreatment: The medicinal materials except Pheretima are dried at 60°C and then made into ultra-fine powder with D90 ≤ 50 μm by air flow pulverization technology, passing through a 300-mesh sieve with a passing rate ≥ 95%.

[0023] Ultrasound-alcohol and water combined extraction: The ultra-fine powder is first added with 6 - 10 times the amount of 30% - 70% ethanol and ultrasonically extracted for 20 - 40 minutes under the conditions of 30 - 50 kHz and 30 - 50°C. Then the medicinal residues are added with 5 - 7 times the amount of water for decoction, each decoction for 1 - 2 hours, and decocted 2 times in total, and the extraction solutions are combined.

[0024] Furthermore, as an improvement to the technical solution of the present invention, when the extraction solution is purified by AB-8 macroporous resin, the concentration of the sample loading solution is 0.2 - 0.4 g crude drug / mL, and the sample is loaded at a flow rate of 3 - 5 BV / h. First, 2 BV of distilled water is used for elution to remove impurities, and then 40% - 60% ethanol is used for elution of 3 - 5 BV to collect the active ingredients.

[0025] Furthermore, as an improvement to the technical solution of the present invention, the concentration process adopts gradient vacuum concentration: First, it is concentrated at 60°C and -0.08 MPa until the relative density at 60°C is 1.15 - 1.20, and then it is heated to 70°C and -0.09 MPa for concentration until the relative density at 60°C is 1.25 - 1.30.

[0026] Furthermore, as an improvement to the technical solution of the present invention, a quality control method for a traditional Chinese medicine formula for traditional Chinese medicine acupuncture: HPLC fingerprint combined with multi-index component determination is adopted:

[0027] The fingerprint needs to contain 9 common characteristic peaks, among which the S3 peak is astragaloside IV and the S7 peak is tanshinone IIA, and their relative peak areas are ≥ 15% and 10% respectively;

[0028] Assay requirements: The content of astragaloside IV (C41H68O14) ≥ 0.8 mg / g, the content of tanshinone IIA (C19H20O3) ≥ 0.5 mg / g, and the content of hydroxysafflor yellow A (C27H32O16) ≥ 1.2 mg / g.

[0029] Furthermore, as an improvement of the technical solution of the present invention, the application of a traditional Chinese medicine formula for traditional Chinese medicine acupuncture in the preparation of drugs for combined acupuncture treatment: The drug is used for the synergistic treatment of pain syndromes of qi stagnation and blood stasis type.

[0030] Through the synergistic optimization of formula design, process innovation and quality control, the present invention solves the technical bottlenecks of existing acupuncture-assisted traditional Chinese medicines in aspects such as component dissolution, transdermal absorption, and preparation adaptation. The specific beneficial effects are as follows:

[0031] 1. Multi-target compound compatibility, breaking through the limitations of single efficacy

[0032] Precise synergistic enhancement of the formula: Astragalus membranaceus - Salvia miltiorrhiza forms a specific qi-tonifying and blood-activating system in a weight ratio of 1:0.75 - 1.25. Detected by HPLC, the dissolution rates of astragaloside IV and tanshinone IIA in this compatibility are increased by 35% and 40% respectively compared with single herbs; safflower is limited to a content of hydroxysafflor yellow A ≥ 2%, and after directional enzymatic hydrolysis of earthworm by trypsin, the proportion of peptides ≤ 5 kDa ≥ 60%. The two work together to promote the transdermal absorption of drugs, and the 24-hour cumulative penetration amount is increased by several times compared with traditional decoctions.

[0033] Process innovation for efficiency improvement: The medicinal materials are pulverized by air flow to D90 ≤ 50 μm, and the cell wall breaking rate reaches more than 95%, increasing the dissolution rates of effective components such as tanshinone IIA and astragaloside IV by 49.2% and 37.4% respectively; the ultrasonic - alcohol - water combined extraction combined with gradient decompression concentration process is adopted, and different extraction conditions are designed for components with different polarities, and the extraction efficiency of lipophilic components is increased by more than 60%, while protecting the thermosensitive component tetramethylpyrazine (retention rate ≥ 85%).

[0034] Diverse preparation adaptation: Develop various dosage forms such as acupoint application gels, enteric-coated tablets, and granules. Among them, the gel uses carbomer 940 matrix combined with 0.3% - 0.7% azone penetration enhancer, and the dynamic viscosity at 25°C is controlled at 1000 - 1500 mPa·s, having both acupoint adhesiveness and drug sustained-release performance; the enteric-coated tablets achieve targeted release in the intestine through coating with acrylic resin II, and the granules use dextrin - lactose excipient (2:1) to ensure the feasibility of industrial production.

[0035] Strict and stable quality control: An HPLC fingerprint containing 9 common characteristic peaks was established, and the content determination methods of index components such as astragaloside IV (≥0.8mg / g) and tanshinone IIA (≥0.5mg / g) were clarified to ensure the consistency of ingredients between batches; accelerated experiments showed that the content of each index component remained above 88% of the initial value within 6 months, and the stability was better than the quality control method required by the "Chinese Pharmacopoeia".

[0036] The clinical efficacy was significant: the total effective rate of the acupuncture combined with the preparation of the present invention treatment group reached 95.0%, and the pain VAS score was reduced to 2.1±0.8, which was significantly better than the acupuncture alone group (77.5%) and the single preparation group (82.5%), confirming its synergistic therapeutic advantage in analgesia and anti-inflammatory. DETAILED DESCRIPTION

[0037] The present invention will be described in detail below with reference to specific embodiments. The illustrative embodiments and descriptions of the present invention are used to explain the present invention but are not intended to limit the present invention.

[0038] A traditional Chinese medicine prescription for acupuncture and moxibustion, comprising the following raw materials in parts by weight:

[0042] The core herbal pair for invigorating Qi: 20-30 parts of Astragalus and 15-25 parts of Salvia miltiorrhiza, with a weight ratio of 1:0.75-1.25;

[0043] Blood circulation and collateral dredging group: 15-25 parts of Chinese Angelica sinensis, 10-20 parts of Chuanxiong, 10-20 parts of Red Peony Root, 10-15 parts of Achyranthes bidentata;

[0044] Transdermal enhancement group: 10-15 parts of safflower, with a hydroxysafflower yellow A content of ≥2%; 10-15 parts of earthworm, with a peptide fraction of ≤5kDa after enzymatic hydrolysis accounting for ≥60%;

[0045] Blending adjuvants: 10-15 parts of peach kernel and 5-10 parts of licorice.

[0046] Specifically, in the present embodiment, the enzymatic hydrolysis treatment of the earthworm is as follows: after freeze-drying at -50°C, the earthworm is dissolved in a buffer solution with a pH of 7.5-8.5 at a mass-to-volume ratio of 1:8 to 1:12, and 1%-2% of the mass of the earthworm is added with trypsin. The earthworm is enzymatically hydrolyzed at 45-55°C for 3-5 hours, and the enzyme is inactivated and then centrifuged to obtain the supernatant for freeze-drying.

[0047] Specifically, in this embodiment, the weight ratio of the Astragalus membranaceus-Danshen miltiorrhiza medicinal pair is 25:20.

[0048] Specifically, in this embodiment, the acupuncture synergistic preparation of the traditional Chinese medicine prescription is an acupoint application gel, which contains the following excipients in the following mass percentages:

[0049] Matrix: Carbomer 940 accounts for 1.0% - 2.0%, glycerol accounts for 4% - 6%, and triethanolamine accounts for 0.3% - 0.7%;

[0050] Penetration enhancer: Azone accounts for 0.3% - 0.7%;

[0051] Extract powder: Based on crude drugs, it accounts for 15% - 25%, and the rest is deionized water.

[0052] Specifically, in the solution of this embodiment, the viscosity of the acupoint application gel is controlled by rheological detection: at 25°C, the dynamic viscosity is 1000 - 1500 mPa·s, and the yield stress ≤ 5 Pa.

[0053] Specifically, in the solution of this embodiment, a preparation method of a traditional Chinese medicine formula for traditional Chinese medicine acupuncture includes the following steps:

[0054] Ultra - fine pulverization pretreatment: The herbs except Pheretima are dried at 60°C and then made into ultra - fine powder with D90 ≤ 50 μm by air - flow pulverization technology, passing through a 300 - mesh sieve with a passing rate ≥ 95%;

[0055] Ultrasound - alcohol - water combined extraction: The ultra - fine powder is first added with 6 - 10 times the amount of 30% - 70% ethanol and ultrasonically extracted for 20 - 40 minutes at 30 - 50 kHz and 30 - 50°C. Then the medicinal residues are added with 5 - 7 times the amount of water and decocted. Each decoction is 1 - 2 hours, and it is decocted 2 times in total. The extraction solutions are combined.

[0056] Specifically, in the solution of this embodiment, when the extraction solution is purified by AB - 8 macroporous resin, the concentration of the sample loading solution is 0.2 - 0.4 g crude drug / mL, and the sample is loaded at a flow rate of 3 - 5 BV / h. First, 2 BV of distilled water is used for elution to remove impurities, and then 40% - 60% ethanol is used for elution of 3 - 5 BV to collect the active ingredients.

[0057] Specifically, in the solution of this embodiment, the concentration process adopts gradient vacuum concentration: First, it is concentrated at 60°C and - 0.08 MPa until the relative density at 60°C is 1.15 - 1.20, and then it is heated to 70°C and - 0.09 MPa for concentration until the relative density at 60°C is 1.25 - 1.30.

[0058] Specifically, in the solution of this embodiment, a quality control method for a traditional Chinese medicine formula for traditional Chinese medicine acupuncture: HPLC fingerprint combined with multi - index component determination is adopted:

[0059] The fingerprint should contain 9 common characteristic peaks. Among them, the S3 peak is astragaloside IV, and the S7 peak is tanshinone IIA. Their relative peak areas are respectively ≥ 15% and 10%;

[0060] Content determination requirements: The content of astragaloside IV (C41H68O14) ≥ 0.8 mg / g, the content of tanshinone IIA (C19H20O3) ≥ 0.5 mg / g, and the content of hydroxysafflor yellow A (C27H32O16) ≥ 1.2 mg / g.

[0061] Specifically, in the solution of this embodiment, the application of a traditional Chinese medicine formula for traditional Chinese medicine acupuncture in the preparation of acupuncture combined treatment drugs: The drug is used for the synergistic treatment of pain caused by qi stagnation and blood stasis.

[0062] Furthermore, it should be noted that for a traditional Chinese medicine formula for traditional Chinese medicine acupuncture, the compatibility principle of the traditional Chinese medicine formula:

[0063] The formula of the present invention follows the traditional Chinese medicine theory of "tonifying qi and promoting blood circulation, dredging collaterals and penetrating the skin", and forms a ternary synergistic system through the compatibility of monarch, minister, assistant, and envoy:

[0064] Compatibility of the monarch drug (core of tonifying qi and promoting blood circulation):

[0065] Astragalus membranaceus (monarch drug): Sweet in taste and slightly warm in nature, belonging to the spleen and lung meridians, mainly tonifying qi and ascending yang, consolidating the exterior and promoting fluid production. Modern pharmacological research shows that it contains components such as astragaloside IV, which can promote qi and blood circulation and improve microcirculation (Experimental Example 1);

[0066] Salvia miltiorrhiza (minister drug): Bitter in taste and slightly cold in nature, belonging to the heart and liver meridians, mainly promoting blood circulation to remove stasis and dredging channels to relieve pain. Its active ingredient, tanshinone IIA, can inhibit inflammatory reactions and regulate the release of neurotransmitters (refer to the following Experimental Example 4).

[0067] Synergy of the drug pair: Astragalus membranaceus - Salvia miltiorrhiza forms a "drug pair for tonifying qi and promoting blood circulation" in a weight ratio of 1:0.75 - 1.25, which conforms to the theory of "qi being the commander of blood, and blood being the mother of qi". Astragalus membranaceus tonifies qi to assist Salvia miltiorrhiza in promoting blood circulation, and Salvia miltiorrhiza promotes blood circulation to assist Astragalus membranaceus in tonifying qi. Their dissolution rates are increased by 35% and 40% respectively compared with single herbs, and they act together on acupuncture-related targets such as TRPV1 and COX-2 to enhance the analgesic and anti-inflammatory effects (refer to the following Experimental Example 1).

[0068] Compatibility of the minister drug (enhancing the effect of dredging collaterals and relieving pain):

[0069] Angelica sinensis and Ligusticum chuanxiong: Angelica sinensis nourishes blood and promotes blood circulation, and Ligusticum chuanxiong promotes qi movement and relieves pain. The combination of the two enhances the efficacy of promoting blood circulation and qi movement and improves local blood circulation;

[0070] Paeonia lactiflora and Achyranthes bidentata: Paeonia lactiflora clears heat and cools blood, disperses stasis and relieves pain, and Achyranthes bidentata removes blood stasis and dredges channels, guiding the drug downward. Together, they dredge the blockage of the meridians and relieve pain caused by qi stagnation and blood stasis (refer to the following Example 2).

[0071] Compatibility of the assistant and envoy drugs (penetrating the skin and regulating harmony):

[0072] Safflower, Earthworm (adjuvant drug): Safflower contains hydroxysafflor yellow A (≥2%), which can dilate blood vessels and improve microcirculation; after enzymatic hydrolysis by trypsin, the peptide segments with a molecular weight ≤5 kDa in earthworm account for ≥60%. Its small molecule peptides can destroy the structure of the skin cutin layer and cooperate with the penetration enhancer azone to increase the 24-hour cumulative penetration amount of the drug by 3.12 times (refer to Experimental Example 2 below);

[0073] Peach Kernel, Licorice (harmonizing drug): Peach Kernel promotes blood circulation and removes blood stasis, and Licorice can harmonize various drugs, relieve spasm and pain, and also has the functions of correcting the taste and stabilizing the preparation. Example 1

[0074] Description of key technical effects

[0075] Improvement of component dissolution efficiency:

[0076] The ultrafine comminution technology (≤50 μm) enables the cell wall breaking rate of medicinal materials to reach more than 95%. The dissolution rates of components such as tanshinone IIA and astragaloside IV are increased by 49.2% and 37.4% respectively, significantly improving the release amount of active ingredients compared with the traditional comminution process (refer to Experimental Example 1 below);

[0077] Ultrasound-alcohol-water combined extraction is used for differential extraction of different polarity components. Lipid-soluble components (such as tanshinone IIA) are extracted by ultrasonic extraction with 30%-70% ethanol, and water-soluble components (such as paeoniflorin) are supplemented by water decoction extraction. The overall extraction efficiency is increased by 60% compared with the single water decoction method (refer to Example 2 below).

[0078] Optimization of transdermal absorption performance:

[0079] The enzymatic hydrolysis product of earthworm contains a large amount of small molecule active peptides (≤5 kDa), which can enhance the fluidity of the skin cutin layer. Combined with 0.3%-0.7% azone penetration enhancer, the transdermal absorption efficiency is increased by more than 3 times, and the 24-hour cumulative penetration amount reaches 589. \(7\ μg / cm\) 2 , solving the problem of low transdermal rate of traditional Chinese medicine external preparations (refer to Experimental Example 2 below);

[0080] The acupoint application gel uses carbomer 940 as the matrix, and the dynamic viscosity at 25°C is controlled at 1000-1500 mPa·s, with appropriate adhesiveness and spreading property, which can closely adhere to the acupoints and continuously release drugs for 12-24 hours (refer to Example 2 below).

[0081] Enhancement of preparation stability and applicability:

[0082] The gradient decompression concentration process (60°C low-temperature concentration → 70°C further concentration) enables the retention rate of the thermosensitive component tetramethylpyrazine to be ≥85%, significantly improving the component stability compared with the traditional high-temperature decoction (loss rate 30%). Preparation method;

[0083] An HPLC fingerprint with 9 common characteristic peaks was established, and the content determination methods for 3 index components were clarified. The accelerated experiment for 6 months showed that the retention rate of component content was ≥88%, higher than the requirement of 85% in the Chinese Pharmacopoeia, ensuring the safety of clinical medication (refer to Experimental Example 3 below).

[0084] The synergistic effect of acupuncture is significant:

[0085] Clinical controlled trials showed that the total effective rate of the treatment group with acupuncture combined with the preparation of the present invention reached 95.0%, and the pain VAS score dropped to 2.1±0.8, showing significant advantages compared with the simple acupuncture group (77.5%) and the single preparation group (82.5%). It was confirmed that the synergistic effect of "acupuncture stimulating acupoints - transdermal drug absorption" could enhance the effects of regulating neurotransmitters and inhibiting inflammatory reactions (refer to Experimental Example 4 below).

[0086] Examples:

[0087] Example 1: Preparation of the basic formula (granules)

[0088] Raw material composition (parts by weight): 20 parts of Astragalus membranaceus, 15 parts of Salvia miltiorrhiza, 15 parts of Angelica sinensis, 10 parts of Ligusticum chuanxiong, 10 parts of Paeonia lactiflora, 10 parts of Achyranthes bidentata, 10 parts of Carthamus tinctorius (the content of hydroxysafflor yellow A is 2.1%), 10 parts of Pheretima aspergillum (the peptide segments ≤5 kDa after enzymatic hydrolysis are 62%), 10 parts of Prunus persica, 5 parts of Glycyrrhiza uralensis

[0089] Preparation steps:

[0090] Pretreatment of Pheretima aspergillum: Pheretima aspergillum was freeze-dried at -50°C for 48 h, dissolved in pH 7.5 phosphate buffer at a ratio of 1:12 (w / v), 1.5% trypsin (calculated based on the mass of Pheretima aspergillum) was added, enzymolyzed at 45°C for 5 h, inactivated at 100°C for 20 min, centrifuged at 10000 rpm for 20 min, and the supernatant was freeze-dried to obtain Pheretima aspergillum peptide powder;

[0091] Ultrafine pulverization: The remaining medicinal materials were dried at 60°C until the water content was 4%, and then pulverized by air flow to D90 = 45 μm (the passing rate through a 300-mesh sieve was 98%);

[0092] Combined extraction: The ultrafine powder was added with 6 times the amount of 30% ethanol, ultrasonic extracted at 30 kHz and 30°C for 40 min, and filtered; the medicinal residues were added with 5 times the amount of water and decocted twice (1 h each time), and the extraction solutions were combined;

[0093] Resin purification: The extraction solution was passed through an AB-8 resin column (loading flow rate 2 BV / h), eluted with 40% ethanol for 5 BV, and the eluate was collected;

[0094] Concentration and granulation: The eluate was concentrated under reduced pressure at 60°C to a relative density of 1.15, dextrin:lactose = 2:1 (the total mass was 3 times that of the extract) was added, granulated and dried to obtain granules.

[0095] Example 2: Optimized formula acupoint application gel (orthogonal test optimization process)

[0096] Raw material composition (parts by weight): 25 parts of Astragalus membranaceus, 20 parts of Salvia miltiorrhiza, 20 parts of Angelica sinensis, 15 parts of Ligusticum chuanxiong, 15 parts of Paeonia lactiflora, 12 parts of Achyranthes bidentata, 12 parts of Carthamus tinctorius (hydroxysafflor yellow A content 2.3%), 12 parts of Pheretima aspergillum (enzymatic hydrolysate peptide segment 65%), 12 parts of Prunus persica, 8 parts of Glycyrrhiza uralensis

[0097] Key process parameters:

[0098] Enzymatic hydrolysis conditions: 50°C, pH 8.0, enzymatic hydrolysis for 4 h, yield of earthworm peptide powder 18.5% (calculated based on earthworm raw material);

[0099] Ultrasonic extraction: Extract with 8 times the amount of 50% ethanol at 40 kHz and 40°C for 30 min, extraction rate of paeoniflorin 87.2%;

[0100] Gel matrix: 1.5% of carbomer 940, 5% of glycerol, 0.5% of azone, pH 6.8, dynamic viscosity at 25°C 1200 mPa·s.

[0101] Example 3: Industrial production of enteric-coated tablets (scale of 1000 tablets)

[0102] Raw material treatment:

[0103] Take 250 g of Astragalus membranaceus, 200 g of Salvia miltiorrhiza and other medicinal materials, and obtain 150 g of extract powder (containing 1250 g of crude drug) according to the process of Example 2;

[0104] Excipients: 150 g of dextrin, 50 g of sodium carboxymethyl starch, enteric coating solution of acrylic resin II (containing 30 g of castor oil);

[0105] Preparation process: Mix the extract powder with dextrin, granulate with 85% ethanol (20-mesh sieve), dry at 60°C for 2 h, add sodium carboxymethyl starch and press tablets (tablet weight 0.5 g), and coat with a coating machine (inlet air temperature 80°C, coating time 40 min).

[0106] Example 4: Preparation of honeyed pills (improvement of traditional dosage form)

[0107] Raw material composition (parts by weight): 30 parts of Astragalus membranaceus, 25 parts of Salvia miltiorrhiza, 25 parts of Angelica sinensis, 20 parts of Ligusticum chuanxiong, 20 parts of Paeonia lactiflora, 15 parts of Achyranthes bidentata, 15 parts of Carthamus tinctorius (hydroxysafflor yellow A content 2.5%), 15 parts of Pheretima aspergillum (≤5 kDa peptide segment 68% after enzymatic hydrolysis), 15 parts of Prunus persica, 10 parts of Glycyrrhiza uralensis

[0108] Preparation steps:

[0109] Enzymatic hydrolysis of Pheretima aspergillum: Dissolve in pH 8.5 phosphate buffer at a ratio of 1:8 (w / v), add 2% trypsin, and perform enzymatic hydrolysis at 55°C for 3 h. After inactivating the enzyme, centrifuge and freeze-dry;

[0110] Drug material treatment: The remaining drug materials are dried at 60°C and then ultrafinely pulverized to D90 ≤ 50 μm, and passed through a 300-mesh sieve.

[0111] Alcohol-water extraction: The ultrafine powder is added with 10 times the amount of 70% ethanol, and ultrasonic extracted at 50 kHz and 50°C for 20 min. The drug residues are added with 7 times the amount of water and decocted twice (2 h each time), and the extraction solutions are combined.

[0112] Concentration and paste formation: The extraction solution is purified by AB-8 resin (eluted with 60% ethanol for 3 BV), and gradient concentrated to a relative density of 1.30 (measured at 60°C) to obtain a thick paste.

[0113] Pill-making process: The thick paste is mixed with refined honey (thick paste: honey = 1:1.5, and the honey is refined at 110 - 115°C), and made into honey pills of 9 g each by a pill-rolling machine, and sealed with a wax coating.

[0114] Key parameters:

[0115] The water content of the refined honey is 18% - 20%, and the relative density is 1.37 (25°C).

[0116] The disintegration time limit of the honey pill is ≤ 60 minutes (in line with the general rules for pills in the Chinese Pharmacopoeia).

[0117] Example 5: Preparation of soft capsule (modern transdermal preparation)

[0118] Raw material composition (parts by weight): 22 parts of Astragalus membranaceus, 18 parts of Salvia miltiorrhiza, 18 parts of Angelica sinensis, 12 parts of Ligusticum chuanxiong, 12 parts of Paeonia lactiflora, 11 parts of Achyranthes bidentata, 11 parts of Carthamus tinctorius (the content of hydroxysafflor yellow A is 2.2%), 11 parts of Pheretima aspergillum (the enzymatically hydrolyzed peptide segment is 63%), 11 parts of Prunus persica, 6 parts of Glycyrrhiza uralensis

[0119] Preparation steps:

[0120] Preparation of the oil phase: Take 20 g of the extract powder (prepared according to the process of Example 2), add 80 g of olive oil and 5 g of lecithin (as an emulsifier), and stir at 50°C until completely dispersed.

[0121] Preparation of the water phase: Mix 90 g of deionized water, 10 g of glycerol, and 0.5 g of azone, and keep warm at 40°C.

[0122] Soft capsule pressing: The dropping method is adopted, the temperature of the oil phase is 60°C, the temperature of the water phase is 50°C, the aperture of the dropping head is 1.2 mm / 1.5 mm, the coolant is liquid paraffin (15°C), and after collecting the soft capsules, the surface oil agent is washed off with 95% ethanol and dried at 40°C for 24 h.

[0123] Quality control: The content of each soft capsule is 1 g, containing 0.6 g of crude drug, and the disintegration time limit is ≤ 30 minutes (simulating the acupoint penetration environment).

[0124] Innovation points:

[0125] Using olive oil as the matrix to enhance the stability of liposoluble components (such as tanshinone IIA);

[0126] Lecithin and azone synergistically promote transdermal absorption. Detected by Franz diffusion cell, the permeation amount of tanshinone IIA in 24 hours is increased by 40% compared with traditional ointment.

[0127] Experimental example 1: Comparative experiment on dissolution rate of active ingredients

[0128] Method: Take the medicinal material powder of Example 2 (traditional pulverization through 80-mesh sieve) and ultrafine powder (≤50μm), and use traditional water decoction method (adding 10 times water, decocting twice for 2 hours each) and the ultrasonic-alcohol-water combined extraction method of the present invention respectively to determine the dissolution rates of tanshinone IIA and astragaloside IV.

[0129]

[0130] Conclusion: The ultrafine pulverization combined extraction process significantly improves the dissolution rate of liposoluble components, and the tanshinone IIA is increased by nearly 50%.

[0131] Experimental example 2: Transdermal absorption kinetics experiment

[0132] Method: Using the improved Franz diffusion cell, with the abdominal skin of rats as the transdermal barrier, compare the 24-hour cumulative permeation amounts of the gel of Example 2 and traditional decoction (containing the same amount of crude drug).

[0133]

[0134]

[0135] Conclusion: The 24-hour cumulative permeation amount of the acupoint application gel is 3.12 times that of the traditional decoction, and the azone penetration enhancer and earthworm peptide powder synergistically enhance the transdermal effect.

[0136] Experimental example 3: Accelerated stability experiment

[0137] Condition: Place the gel of Example 2 in an environment of 40°C ± 2°C and RH75% ± 5%, and regularly detect the content changes of the index components.

[0138]

[0139] Conclusion: The contents of each component remain above 88% of the initial value within 6 months, meeting the stability requirements of the Chinese Pharmacopoeia.

[0140] Experimental example 4: Clinical efficacy comparative trial

[0141] Case selection: Select 120 patients with chronic lumbar muscle strain of qi stagnation and blood stasis type, and randomly divide them into 3 groups (40 cases in each group):

[0142] · Group A: Simple acupuncture treatment (once a day, 10 times as a course of treatment);

[0143] · Group B: Gel application of the present invention (changed every 12 h, with the same course of treatment as above);

[0144] · Group C: Acupuncture combined with gel application (superposition of treatment methods).

[0145] Efficacy evaluation:

[0146] · Visual analogue scale (VAS) for pain: After treatment, the VAS score of Group C (2.1 ± 0.8) was significantly lower than that of Group A (3.9 ± 1.2) and Group B (3.2 ± 1.0) (P < 0.01);

[0147] · Total clinical effective rate: 95.0% (38 / 40) in Group C, 82.5% (33 / 40) in Group B, 77.5% (31 / 40) in Group A. There was a statistically significant difference between Group C and Group A (χ 2 = 6.83, P < 0.01).

[0148] In summary, through the collaborative optimization of formula design, process innovation and quality control, the present invention solves the technical bottlenecks of existing acupuncture-assisted traditional Chinese medicine in aspects such as component dissolution, transdermal absorption and preparation adaptation. The specific beneficial effects are as follows:

[0149] 1. Multi-target compound compatibility, breaking through the limitation of single efficacy

[0150] Precise efficacy enhancement of the formula: Astragalus membranaceus - Salvia miltiorrhiza forms a specific qi-tonifying and blood-activating system in a weight ratio of 1:0.75 - 1.25. Detected by HPLC, the dissolution rates of astragaloside IV and tanshinone IIA in this compatibility are increased by 35% and 40% respectively compared with single herbs; safflower is limited to a content of hydroxysafflor yellow A ≥ 2%, and the proportion of peptides ≤ 5 kDa in earthworm after directional enzymatic hydrolysis by trypsin is ≥ 60%. The two work together to promote the transdermal absorption of drugs, and the cumulative penetration amount in 24 h is increased several times compared with traditional decoctions.

[0151] Process innovation for efficiency improvement: The medicinal materials are pulverized by air flow to D90 ≤ 50 μm, and the cell wall breaking rate reaches more than 95%, so that the dissolution rates of effective components such as tanshinone IIA and astragaloside IV are increased by 49.2% and 37.4% respectively; the ultrasonic-alcohol-water combined extraction combined with gradient decompression concentration process is adopted, and different extraction conditions are designed for different polarity components. The extraction efficiency of lipophilic components is increased by more than 60%, and at the same time, the thermosensitive component tetramethylpyrazine is protected (retention rate ≥ 85%).

[0152] Multi-adaptable formulations: We have developed a variety of dosage forms, including acupoint patch gels, enteric-coated tablets, and granules. The gel uses a carbomer 940 matrix combined with 0.3%-0.7% azone permeation enhancer, with a dynamic viscosity at 1000-1500 mPa·s at 25°C, combining acupoint adhesion with sustained-release drug properties. Enteric-coated tablets are coated with acrylic resin No. II to achieve targeted intestinal release, and granules use dextrin-lactose excipients (2:1) to ensure the feasibility of industrial production.

[0153] Strict and stable quality control: An HPLC fingerprint containing 9 common characteristic peaks was established, and the content determination methods of index components such as astragaloside IV (≥0.8mg / g) and tanshinone IIA (≥0.5mg / g) were clarified to ensure the consistency of ingredients between batches; accelerated experiments showed that the content of each index component remained above 88% of the initial value within 6 months, and the stability was better than the quality control method required by the "Chinese Pharmacopoeia".

[0154] The clinical efficacy was significant: the total effective rate of the acupuncture combined with the preparation of the present invention treatment group reached 95.0%, and the pain VAS score was reduced to 2.1±0.8, which was significantly better than the acupuncture alone group (77.5%) and the single preparation group (82.5%), confirming its synergistic therapeutic advantage in analgesia and anti-inflammatory.

[0155] The technical solutions provided by the embodiments of the present invention are introduced in detail above. Specific examples are used herein to illustrate the principles and implementation methods of the embodiments of the present invention. The description of the above embodiments is only applicable to help understand the principles of the embodiments of the present invention. At the same time, for those skilled in the art, according to the embodiments of the present invention, there may be changes in the specific implementation methods and application scopes. In summary, the contents of this specification should not be understood as limiting the present invention.

Claims

1. A traditional Chinese medicine prescription for traditional Chinese medicine acupuncture and moxibustion, characterized in that, It consists of raw materials in the following parts by weight: Qi-tonifying core herb pair: 20 - 30 parts of Astragalus membranaceus and 15 - 25 parts of Salvia miltiorrhiza, and the weight ratio of the two is 1:0.75 - 1.25; Blood-activating and collaterals-unblocking group: 15 - 25 parts of Angelica sinensis, 10 - 20 parts of Ligusticum chuanxiong, 10 - 20 parts of Paeonia lactiflora, 10 - 15 parts of Achyranthes bidentata; Transdermal efficacy-enhancing group: 10 - 15 parts of Carthamus tinctorius, and the content of hydroxysafflor yellow A is ≥2%; 10 - 15 parts of Pheretima aspergillum, and the proportion of peptide segments with a molecular weight ≤5 kDa after enzymatic hydrolysis is ≥60%; Harmonizing and adjuvant herbs: 10 - 15 parts of Prunus persica and 5 - 10 parts of Glycyrrhiza uralensis.

2. The traditional Chinese medicine prescription for traditional Chinese medicine acupuncture according to claim 1, wherein: The enzymatic hydrolysis treatment of the Pheretima aspergillum is as follows: After freeze-drying at -50°C, it is dissolved in a buffer solution with a mass-to-volume ratio of 1:8 to 1:12 at pH 7.5 - 8.5, 1% - 2% of trypsin based on the mass of Pheretima aspergillum is added, and enzymatic hydrolysis is carried out at 45 - 55°C for 3 - 5 hours. After inactivating the enzyme, the supernatant is centrifuged and freeze-dried.

3. A traditional Chinese medicine prescription for traditional Chinese medicine acupuncture according to claim 1, characterized in that: The weight ratio of the Astragalus membranaceus - Salvia miltiorrhiza herb pair is 25:

20.

4. A traditional Chinese medicine prescription for traditional Chinese medicine acupuncture according to claim 1, characterized in that: The acupuncture and moxibustion synergistic preparation of the traditional Chinese medicine prescription is a transdermal patch gel, and the transdermal patch gel contains the following excipients in mass percentage: Matrix: Carbomer 940 accounts for 1.0% - 2.0%, glycerol accounts for 4% - 6%, triethanolamine accounts for 0.3% - 0.7%; Penetration enhancer: Azone accounts for 0.3% - 0.7%; Extract powder: Calculated by crude drug, it accounts for 15% - 25%, and the rest is deionized water.

5. A traditional Chinese medicine prescription for traditional Chinese medicine acupuncture according to claim 1, characterized in that: The transdermal patch gel controls the viscosity of the preparation by rheological detection: At 25°C, the dynamic viscosity is 1000 - 1500 mPa·s, and the yield stress ≤5 Pa.

6. The preparation method of the traditional Chinese medicine prescription according to any one of claims 1-3, characterized in that, It includes the following steps: Ultra-fine pulverization pretreatment: The herbs except Pheretima aspergillum are dried at 60°C and then made into ultra-fine powder with D90 ≤50 μm by air-flow pulverization technology, passing through a 300-mesh sieve with a passing rate ≥95%; Ultrasound - ethanol-water combined extraction: The ultra-fine powder is first added with 6 - 10 times the amount of 30% - 70% ethanol, and ultrasonic extraction is carried out at 30 - 50 kHz and 30 - 50°C for 20 - 40 minutes. The medicinal residues are then added with 5 - 7 times the amount of water for decoction, each decoction for 1 - 2 hours, and decocted 2 times in total, and the extraction solutions are combined.

7. The preparation method of the traditional Chinese medicine prescription according to claim 6, characterized in that: When the extraction solution is purified by AB-8 macroporous resin, the concentration of the loading solution is 0.2 - 0.4 g of crude drug / mL, and the loading is carried out at a flow rate of 3 - 5 BV / h. First, 2 BV of distilled water is used for elution to remove impurities, and then 40% - 60% ethanol is used for elution of 3 - 5 BV to collect the active ingredients.

8. The preparation method of the traditional Chinese medicine prescription according to claim 6, characterized in that: The concentration process adopts gradient vacuum concentration: First, it is concentrated at 60°C and -0.08 MPa until the relative density at 60°C is 1.15 - 1.20, and then it is heated to 70°C and -0.09 MPa for concentration until the relative density at 60°C is 1.25 - 1.

30.

9. The quality control method of the traditional Chinese medicine prescription according to any one of claims 1-3, characterized in that HPLC fingerprint combined with multi-index component determination is adopted: The fingerprint needs to contain 9 common characteristic peaks, among which the S3 peak is astragaloside IV and the S7 peak is tanshinone IIA, and their relative peak areas are respectively ≥15% and 10%; Assay requirements: The content of astragaloside IV (C41H68O14) ≥ 0.8 mg / g, the content of tanshinone IIA (C19H20O3) ≥ 0.5 mg / g, and the content of hydroxysafflor yellow A (C27H32O16) ≥ 1.2 mg / g.

10. Use of the traditional Chinese medicine prescription according to any one of claims 1-3 in the preparation of a drug for combined acupuncture and moxibustion treatment, characterized in that, The said drug is used for the synergistic treatment of pain syndromes due to qi stagnation and blood stasis.

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