A granular medicine of erdun-7 and a preparation method thereof

Abstract

This invention relates to the field of Mongolian medicine preparation technology, and discloses an Erdun-7 granule preparation and its preparation method, which is made from the following raw materials in parts by weight: 1 part of *Inula japonica* and 1.3-1.8 parts of *Rubus idaeus*, used to regulate qi, relieve exterior symptoms and reduce fever; 1.6-2.4 parts of *Sophora flavescens*, 0.7-1.3 parts of *Gardenia jasminoides*, and 0.7-1.3 parts of *Terminalia chebula*, used to enhance the heat-clearing, fire-purging, detoxifying and blood-cooling effects of *Inula japonica* and *Rubus idaeus*; 0.8-1.2 parts of *Uncaria rhynchophylla* and 0.8-1.2 parts of *Cicada slough*, used to calm the liver, extinguish wind, and stop spasms; and 0.7-1.3 parts of *Kaempferia galanga* and 0.7-1.3 parts of *Melia azedarach*. This Erdun-7 granule preparation and its preparation method target the anticonvulsant effect of the granules, prolonging the latency period and reducing the incidence and severity, which is significantly better than traditional preparations, demonstrating the synergistic effect of clearing heat and relieving exterior symptoms and extinguishing wind and stopping spasms.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine preparation technology, and in particular to an Erdun-7 granule preparation and its preparation method. Background Technology

[0002] Erdun-7 Decoction is a classic compound formula in Mongolian medicine for treating fever and the early stages of epidemics. Its traditional formula consists of seven medicinal materials, including costus root, raspberry wood (pearl stem), and sophora flavescens. It has clear effects of clearing heat, detoxifying, and relieving pain. In existing technology, it has been made into ordinary granules for convenient administration. Its preparation process is mostly the conventional method of decocting all the medicinal materials together, concentrating, adding excipients, and granulating.

[0003] However, when this traditional formula is used to prevent and treat the specific and critical clinical condition of febrile seizures in children, firstly, the original formula is not very targeted in calming wind and stopping spasms, and its preventive and control effects on seizures have not been optimized or verified; secondly, the herbs in the formula, such as Sichuan pepper and sophora flavescens, have certain toxicity or strong bitter and cold properties, and using them raw or in conventional decoction poses potential risks to children's delicate spleen, stomach, liver, and kidney functions; thirdly, traditional granules lack targeting in the gastrointestinal tract, and the drug is easily released in the stomach, which may not only aggravate irritation but also prevent the newly added prebiotics and other modern active ingredients in the formula from being accurately delivered to the intestinal target area to exert their effects; finally, the fixed dosage form and taste are also difficult to meet the special medication adherence needs of pediatric patients. Therefore, developing a safe, efficient, and pediatric-suitable modified Erdon-7 formulation is a technical problem that urgently needs to be solved in this field. Summary of the Invention

[0004] The technical problem to be solved by this invention is that the existing technology has the disadvantage of causing febrile seizures in children. To address this, we propose an Erdun-7 granule preparation and its preparation method.

[0005] To achieve the above objectives, this application adopts the following technical solution: an Erdun-7 granule preparation, comprising the following raw materials in parts by weight: 1 part of *Saussurea costus* and 1.3-1.8 parts of *Rubus idaeus*, used to regulate qi, relieve exterior symptoms and reduce fever; 1.6-2.4 parts of *Sophora flavescens*, 0.7-1.3 parts of *Gardenia jasminoides*, and 0.7-1.3 parts of *Terminalia chebula*, used to enhance the heat-clearing, fire-purging, detoxifying, and blood-cooling effects of *Saussurea costus* and *Rubus idaeus*; and 0. 8-1.2 parts of cicada molting and 0.8-1.2 parts of galangal and Sichuan pepper are used to calm the liver and extinguish wind, and to relieve convulsions and spasms. 0.7-1.3 parts of galangal and 0.7-1.3 parts of Sichuan pepper are used to promote qi circulation, relieve pain, soothe the liver and clear heat, and regulate qi and blood to help the medicine exert its effects. 0.8-1.2 parts of cynomorium and 0.8-1.2 parts of sea buckthorn are used to benefit the essence and strengthen the spleen. Cynomorium is used to benefit the essence and nourish the blood. 3.0-6.0 parts of rock sugar and 3.0-6.0 parts of white sugar. The Erdon-7 granules are coated with a double layer consisting of a protective layer and a targeting layer. The protective layer is composed of chitosan and sodium alginate, which maintains structural integrity in the acidic environment of the stomach and dissolves in the alkaline environment of the intestine. The targeting layer is a bile acid-modified polylactic-co-glycolic acid copolymer (PLGA), which enables the active delivery of the Erdon-7 granules to the target area in the intestine through the specific binding of bile acids to bile acid receptors in the intestine.

[0006] Preferably, the Cynomorium songaricum and the Hippophae rhamnoides are added to the Erdun-7 granule preparation in the form of extracts, and the Cynomorium songaricum polysaccharide in Cynomorium songaricum and the Hippophae rhamnoides oligosaccharide in Hippophae rhamnoides are used to regulate the intestinal flora.

[0007] A method for preparing Erdun-7 granules includes the following steps: Step 1: Provide raw materials: The raw materials include costus root, sophora flavescens, gardenia, chebula, uncaria, cicada slough, galangal, chinaberry, cynomorium, sea buckthorn, rock sugar, and white sugar; First, take Sichuan chinaberry and process it with vinegar; take sophora flavescens and process it with charcoal; take costus root and grind it into coarse powder for later use; then mix the remaining medicinal materials, including gardenia, chebula, uncaria, cicada slough, and galangal. Step 2: Mix the processed products obtained from Sichuan chinaberry and sophora flavescens in Step 1, and perform the first round of extraction. Concentrate the extract for later use. Step 3: Next, combine the remaining medicinal materials from Step 1 with the coarse powder of *Inula japonica* and carry out the second round of extraction. The coarse powder of *Inula japonica* is added 30-60 minutes before the end of the second round of extraction. Step 4: Extraction of Cynomorium songaricum and Hippophae rhamnoides, yielding the first extracts Cynomorium songaricum polysaccharide and Hippophae rhamnoides oligosaccharide; Step 5: Concentrate the extract from Step 2 into a thick paste, mix it with Cynomorium songaricum polysaccharide, Hippophae rhamnoides oligosaccharide, rock sugar and white sugar and granulate to obtain uncoated Erdun-7 granules. Step Six: Sequentially coat the uncoated Erdun-7 granules with a protective layer and a targeting layer. The protective layer is formed by alternately spraying chitosan solution and sodium alginate solution and then ionically crosslinking them. The targeting layer is formed by spraying a cholic acid-polylactic acid-glycolic acid copolymer solution.

[0008] Preferably, for every 100 parts by weight of Sichuan chinaberry, 20-30 parts by weight of rice vinegar are mixed evenly, allowed to soak for 1-2 hours, and then stir-fried over low heat at 80-120℃ for 15-30 minutes until the surface color of the medicinal material deepens and the vinegar smell dissipates.

[0009] Preferably, the charring process involves placing the Sophora flavescens slices in a frying pan at 180-220℃ and stir-frying them rapidly over high heat for 5-10 minutes until the surface of the herb is charred black and the inside is charred brown, while controlling the weight loss rate of the herb within the range of 15%-25%. Then, spray the herb with water at 10%-15% of its weight to extinguish the smoke, and remove it to dry.

[0010] Preferably, the extraction time in the first round of step two is 1.5-2.5 hours, and the extraction time in the second round of step three is 0.5-1.5 hours; wherein the extraction time in the first round is at least 0.5 hours longer than the extraction time in the second round.

[0011] Preferably, when alternately spraying the chitosan solution and the sodium alginate solution, the chitosan solution is sprayed first, and after drying, the sodium alginate solution is sprayed.

[0012] Preferably, the coating weight gain of the protective layer is 3%-5%, and the coating weight gain of the targeting layer is 5%-8%.

[0013] The technical effects and advantages of this invention are as follows: In this invention, Uncaria rhynchophylla and Cicadae Periostracum are added to enhance the effects of calming the liver, extinguishing wind, and relieving convulsions. This makes the entire formula primarily indicated for febrile seizures in children. At the same dosage, the anticonvulsant effect of the targeted granules of this invention, in terms of prolonging the latency period and reducing the incidence and severity, is significantly superior to traditional granules, demonstrating the synergistic effect of clearing heat and relieving exterior symptoms and extinguishing wind and relieving convulsions. Furthermore, the inclusion of vinegar-processed Melia toosendan, charred Sophora flavescens, and Aucklandia lappa significantly reduces the content of toxic components such as azadirachtin and greatly alleviates the gastric mucosal irritation of Sophora flavescens, greatly improving the safety of medication for children while retaining the core efficacy.

[0014] In this invention, the double-layer coating enables the granules to have pH-dependent intestinal targeted release characteristics, which not only effectively protects the stomach from irritation, but also ensures the targeted release and efficient utilization of the drug and prebiotic components such as Cynomorium songaricum polysaccharide and Hippophae rhamnoides oligosaccharide in the intestine, thereby regulating the flora to exert an anticonvulsant effect. Finally, the well-coated granules have uniform particle size, and the taste is improved due to the sugar excipients and coating, making them easy for children to take and carry. Attached Figure Description

[0015] The disclosure of this invention is illustrated with reference to the accompanying drawings. It should be understood that the drawings are for illustrative purposes only and are not intended to limit the scope of protection of this invention. In the drawings, the same reference numerals are used to refer to the same parts: Figure 1 This is a graph showing the release rate of geniposide from the three types of particles of the present invention in a simulated gastrointestinal environment. Figure 2 This is a comparison chart of the incidence of seizures in rats according to the present invention; Figure 3This is a comprehensive comparison chart of the main pharmacodynamic indicators of febrile seizures in rats according to the present invention; Figure 4 This is a graph showing the relative abundance changes of beneficial bacteria genera in this invention. Detailed Implementation

[0016] It is readily understood that, based on the technical solution of this invention, those skilled in the art can propose various interchangeable structural methods and implementations without altering the essential spirit of the invention. Therefore, the following detailed embodiments and accompanying drawings are merely illustrative examples of the technical solution of this invention and should not be considered as the entirety of the invention or as limitations or restrictions on the technical solution of this invention.

[0017] Example 1 An Erdun-7 granule preparation is made from the following raw materials in parts by weight: One part of Aucklandia lappa and 1.3-1.8 parts of Rubus idaeus are used to regulate Qi, relieve exterior symptoms, and reduce fever. Sophora flavescens 1.6-2.4 parts, Gardenia jasminoides 0.7-1.3 parts, Terminalia chebula 0.7-1.3 parts are used to enhance the heat-clearing, fire-purging, detoxifying, and blood-cooling effects of Aucklandia lappa and Rubus idaeus. Uncaria rhynchophylla 0.8-1.2 parts and Cicadae periostracum 0.8-1.2 parts are used to calm the liver and extinguish wind, calm fright and stop spasms. Kaempferia galanga 0.7-1.3 parts and Melia toosendan 0.7-1.3 parts are used to promote qi circulation and relieve pain, soothe the liver and clear heat, regulate qi and blood to help the medicine exert its effects.

[0018] In addition, the following extracts are needed: 0.8-1.2 parts of Cynomorium polysaccharide extract and 0.8-1.2 parts of Hippophae rhamnoides oligosaccharide extract, both of which are refined powders that meet relevant standards, used to benefit the essence and strengthen the spleen. Cynomorium is used to benefit the essence and nourish the blood.

[0019] Use 3.0-6.0 parts rock sugar and 3.0-6.0 parts white sugar. Grind them through a 100-mesh sieve before use.

[0020] First, weigh 100 parts by weight of Sichuan pepper (Chuanlianzi) slices, add 25 parts by weight of rice vinegar (total acidity ≥ 3.5g / 100mL), stir well, seal and let it soak for 1.5 hours to allow the vinegar to fully penetrate. Place the soaked herbs in a wok and stir-fry over low heat for 20 minutes, controlling the wok temperature at 90±10℃. Observe during the stir-frying process; the processing endpoint is when the surface of the herbs turns dark yellow-brown, the texture becomes tough to the touch, and no pungent acetic acid odor is emitted. After processing, remove and spread out to cool. The acetic acid heating reaction reduces the content of potentially hepatotoxic components in Sichuan pepper (Chuanlianzi).

[0021] Next, weigh 100 parts by weight of Sophora flavescens slices, preheat the wok to 200±10℃, add Sophora flavescens, and stir-fry quickly until the surface of the slices is charred and black, the inside is charred brown, and a small amount of smoke is produced. Immediately measure the weight loss rate. When the weight loss rate reaches 20%±5%, quickly spray 15 parts by weight of clean water to extinguish the smoke, and stir-fry quickly to allow the steam to escape. Then, take out the Sophora flavescens and place it in a clean container to cool. The carbonization process destroys the extremely bitter and cold nature of Sophora flavescens, reducing the stimulation to the spleen and stomach of children.

[0022] Next, weigh out the costus root and grind it into a coarse powder that can pass through a 10-mesh sieve using a grinder, then store it separately for later use. Finally, mix the Rubus idaeus, Kaempferia galanga, Terminalia chebula, Gardenia jasminoides, Uncaria rhynchophylla, and Cicadae periostracum evenly for later use.

[0023] The prepared Sichuan pepper seeds, along with a mixture of bitter ginseng, rhubarb, galangal, chebula, gardenia, uncaria, and cicada slough, are placed into a multi-functional extraction tank. Ten times the volume of drinking water is added, and the mixture is heated to a gentle boil for the first round of decoction extraction, which is controlled to last for 2 hours. After extraction, the liquid is filtered out, and the residue is retained.

[0024] Add eight times the amount of drinking water to the dregs and heat for a second round of decoction extraction. One hour after the extraction begins, add the coarse powder of *Saussurea costus* to the container and continue extraction for another 0.5 hours, controlling the actual extraction time of *Saussurea costus* to 0.5 hours. This shortens the heating time of volatile and potentially uncomfortable components in *Saussurea costus*, preserving its aromatic and spleen-invigorating effects while reducing the risk of side effects.

[0025] The first and second extracts were combined and filtered through a 200-mesh filter cloth. The filtrate was transferred to a vacuum concentration tank and concentrated to a thick paste with a relative density of 1.25-1.3 at 60°C under conditions of 65-70°C and -0.08 to -0.10 MPa.

[0026] Next, transfer the above-mentioned thick paste to a mixer, and add in sequence the following: Cynomorium songaricum polysaccharide extract (polysaccharide content ≥50%), Hippophae rhamnoides oligosaccharide extract (oligosaccharide content ≥30%), rock sugar powder, and white sugar powder. Turn on the mixer and stir thoroughly until homogeneous. Add an appropriate amount of 70% ethanol as a wetting agent to obtain a soft material.

[0027] Granulation was performed using a gyratory granulator, passing the granules through a 14-mesh sieve. The wet granules were then placed in a hot air circulating drying oven and dried at 55°C until the moisture content was ≤5.0%. The dried granules were then passed through an 80-mesh sieve using a granulator to obtain uncoated granule cores, i.e., uncoated Erdun-7 granules. By adding Cynomorium songaricum polysaccharide and Hippophae rhamnoides oligosaccharide, not only are they active ingredients, but their mild viscosity also helps in shaping and provides prebiotic effects in the intestines.

[0028] Next, the coating material was prepared, which consisted of a protective layer and a targeting layer. The protective layer was composed of chitosan solution and sodium alginate solution. The chitosan solution used was pharmaceutical-grade chitosan with a deacetamide degree ≥85% and a viscosity of 50-200 mPa·s (1% acetic acid solution, 20℃). 10.0 g was weighed and slowly added to 990 mL of 1% (v / v) acetic acid aqueous solution, and magnetically stirred until completely dissolved to obtain a 1.0% (w / v) chitosan-acetic acid stock solution, which was filtered through a 0.45 μm microporous membrane before use.

[0029] The sodium alginate solution uses pharmaceutical-grade sodium alginate, conforming to the standards of the Chinese Pharmacopoeia. Weigh 10.0 g and dissolve it in 990 mL of purified water, stirring magnetically until completely dissolved to obtain a 1.0% (w / v) sodium alginate aqueous solution. Filter through a 0.45 μm microporous membrane before use.

[0030] The targeting layer is a solution of cholic acid-polylactic acid-glycolic acid copolymer (PLGA). Cholic acid-PLGA is an amphiphilic functional material formed by chemically coupling cholic acid molecules to PLGA polymer chains. The molar ratio of lactic acid to glycolic acid in the PLGA fragment is 75:25, the intrinsic viscosity is 0.2-0.4 dL / g, and the corresponding weight-average molecular weight is 15,000-25,000 Da. To prepare the solution, 2.00 g of the cholic acid-PLGA copolymer is weighed and placed in an appropriate amount of anhydrous dichloromethane. The solution is vortexed until completely dissolved, and then diluted to 100 mL in a volumetric flask to obtain a clear and homogeneous coating working solution with a concentration of 2.0% (w / v).

[0031] In the coating process, a fluidized bed bottom spray coating machine is used, and the entire process is divided into two stages: The first stage involves coating with a protective layer. First, 100g of uncoated Erdun-7 granules is placed in the coating chamber. Key parameters are set: inlet air temperature 45±2℃, material temperature maintained at 38-42℃, fluidizing airflow to ensure uniform suspension of the uncoated Erdun-7 granules, and spray air pressure 0.3-0.4MPa. Next, chitosan solution is sprayed at a rate of 3-5mL / min for 30 seconds, then stopped. Hot air at 45℃ is then introduced for drying and curing for 3 minutes to ensure uniform chitosan adhesion. Subsequently, sodium alginate solution is switched to and sprayed at the same rate for 30 seconds, followed by another 3 minutes of hot air drying and curing. When the positively charged chitosan and negatively charged sodium alginate come into contact on the particle surface, in-situ ionic cross-linking occurs, forming a dense gel network. Finally, repeat the above cycle, pausing after every 5 cycles, taking out a small sample and weighing it precisely. When the weight gain of the coating reaches 4.0 ± 0.5% of the initial weight of the uncoated Erdun-7 granules, stop the first stage of coating. At this point, a complete, pH-dependent hydrogel protective layer is formed on the surface of the granules.

[0032] The second stage involves coating the target layer. The particles coated with the protective layer from the first stage are not removed. The fluidized bed inlet air temperature is adjusted to 40±2℃ to suit the organic solvent. The process is switched to a cholic acid-PLGA solution for spraying, with the spray rate controlled at 2-4 mL / min to avoid solvent overload and particle adhesion. An intermittent spray mode is used to allow the organic solvent (dichloromethane) to fully evaporate during fluidized drying, allowing PLGA polymers to deposit layer by layer. Spraying is continued and weighed periodically. Spraying is stopped when the coating weight gain reaches 6.5±1.0% of the weight of the protective layer-coated particles. This cholic acid-PLGA layer forms on the outermost layer, with the cholic acid molecules acting as targeting ligands exposed on the outermost side of the particles.

[0033] After coating, maintain fluidization, adjust the inlet air temperature to 35℃, and continue drying for 30 minutes to ensure complete evaporation of dichloromethane and full curing and stabilization of the coating film. Turn off the equipment, remove the particles, and place them in a desiccator at room temperature to equilibrate moisture for at least 2 hours, finally obtaining double-coated targeted Erdun-7 particles, i.e., double-coated Erdun-7 particles.

[0034] The resulting double-coated Erdun-7 particles exhibited a release rate of <10% within 2 hours in simulated gastric fluid (pH 1.2); upon transfer to simulated intestinal fluid (pH 6.8), the release rate was >80% within 45 minutes, with an angle of repose ≤30°. Structurally, the drug is composed of standard intermediates, containing all pharmacodynamic components and prebiotics; the protective layer consists of chitosan and sodium alginate ion-crosslinked gel; and the targeting layer is composed of a bile acid-PLGA polymer membrane.

[0035] To verify the above preparation process and the efficacy of the obtained Erdun-7 granules, the following in vitro and in vivo experiments were conducted.

[0036] Experiment 1: Verification of in vitro release rate Experimental materials and instruments Samples: conventional Erdun-7 granules (TG) prepared in Comparative Example 1, uncoated Erdun-7 granules (ICG) prepared in Example 1, and double-coated Erdun-7 granules (ICG-C) prepared in Example 1. Each batch of samples was used after equilibration of moisture in a desiccator for one week.

[0037] Reagents: hydrochloric acid, potassium dihydrogen phosphate, sodium hydroxide; the water used in the experiment was ultrapure water.

[0038] Medium: Simulated gastric juice (SGF): 0.1 mol / L HCl solution, pH 1.20 ± 0.05; Simulated intestinal fluid (SIF): 0.05 mol / L phosphate buffer, pH 6.80 ± 0.05.

[0039] Instrument: RC-806 dissolution tester, equipped with an automatic sampling system; Agilent 1260 Infinity II high-performance liquid chromatography system, equipped with a DAD detector and ChemStation workstation; ME204E analytical balance; pHS-3C pH meter.

[0040] Weigh appropriate amounts of each sample powder, and calculate the total amount of geniposide in the dosage to be equivalent to 25.0 mg based on the actual content of geniposide in each sample. Place the powder in 900 mL of dissolution medium. Each group of experiments is repeated in 6 replicates (n=6). According to the second method (paddle method) in General Chapter 0931 of the Chinese Pharmacopoeia (2025 edition) for the determination of dissolution and release, the rotation speed is controlled at 50±1 rpm and the temperature is 37.0±0.5℃.

[0041] The experiment was conducted in 900 mL of simulated gastric juice (SGF) over 0–2 hours.

[0042] At 2 hours, an equal volume (900 mL) of preheated 37°C simulated intestinal fluid (SIF) stock solution with twice the concentration was quickly added to each dissolution vessel to instantly adjust the pH of the system to 6.80, and the total volume became 1800 mL, and the experiment continued.

[0043] At time points of 0.5 hours, 1 hour, 1.5 hours, 2 hours, 2.25 hours, 2.5 hours, 3 hours, 4 hours, and 6 hours, 5 mL of blank medium at the same temperature and pH was automatically sampled and filtered immediately through a 0.45 μm syringe filter. Immediately after sampling, 5 mL of blank medium at the same temperature and pH was added to the dissolution vessel.

[0044] The content of geniposide was determined using a validated high-performance liquid chromatography (HPLC) method, with the specific chromatographic conditions as follows: The chromatographic system was configured with a Waters XSelect HSS C18 column (4.6×250 mm, 5 μm particle size), using acetonitrile as mobile phase A and 0.1% phosphoric acid aqueous solution as mobile phase B, with gradient elution. The proportion of mobile phase A was linearly increased from 10% to 20% within 0-15 minutes, and the flow rate was kept constant at 1.0 mL / min. The column oven temperature was set to 30℃. A diode array detector was used for detection at a wavelength of 238 nm. The injection volume was 10 μL.

[0045] This method uses geniposide reference standard (batch number 110749-202118) provided by the National Institutes for Food and Drug Control for quantification. Validation showed that geniposide exhibits good linearity in the concentration range of 0.5-50 μg / mL.

[0046] Cumulative release Calculate using the formula: ; in, Let be the concentration sampled at time t. For the volume of the medium, This represents the theoretical total amount of geniposide in the sample. For the first The drug concentration measured at each sampling point, This represents the volume of each sample taken.

[0047] Reference Figure 1 This experiment used geniposide as the indicator component to simulate drug release behavior in gastric and intestinal fluids. The cumulative release data of geniposide from the three types of particles in the simulated gastrointestinal environment are shown in Table 1. Table 1 shows the cumulative release rate of geniposide from the three types of particles in a simulated gastrointestinal environment:

[0048] The results showed that ICG-C had an extremely low release rate (<7%) in simulated gastric fluid within 2 hours, demonstrating that its double-layer coating effectively protects the contents. After the medium transitioned to intestinal fluid, its release behavior changed dramatically, with over 70% released within 45 minutes. This contrasts sharply with the rapid release of TG and ICG in gastric fluid, directly and conclusively proving that the double-layer coating of this invention achieves precise intestinal localization and targeted release.

[0049] Experiment 2: Pharmacodynamic Verification of Antipyretics for Febrile Seizures in Children Experimental animals: SPF-grade SD rat pups, 14-16 days old, weighing 32-38g, half male and half female. The animals were acclimatized for 3 days in an SPF-grade environment with a temperature of 22±2℃, humidity of 50±10%, and a cycle of 12 hours of light and 12 hours of no light. They had free access to food and water.

[0050] Rats were intraperitoneally injected with lipopolysaccharide (LPS, Sigma L2880, 200 μg / kg, dissolved in sterile saline). 2.5 hours after injection, rats were individually placed in a transparent plexiglass observation box and placed in a constant temperature water bath oven at 40.0±0.5℃ to induce seizures.

[0051] The 14 young mice were randomly divided into 7 groups (n=2): G1 was the normal control group, injected with an equal volume of physiological saline, and not heated, serving as a control for normal physiological state. G2 served as the model control group, and the model was established by gavage with pure water. G3 is the traditional granule group, which is a traditional Erdun-7 granule (TG) suspension (2.0g crude drug / kg) administered by gavage during modeling. G4 was the uncoated granule group, and the model was established by oral administration of uncoated Erdun-7 granule (ICG) suspension (2.0 g crude drug / kg). G5 was the low-dose targeted particle group, which was modeled by oral administration of double-coated Erdun-7 granules (ICG-C) suspension (2.0g crude drug / kg). G6 was the high-dose targeted particle group, which was used to induce a model by oral administration of a double-coated Erdun-7 granule (ICG-C) suspension (4.0 g crude drug / kg). G7 was the positive control diazepam group. The model was established by intraperitoneal injection of diazepam 0.5 mg / kg. All oral medications were freshly prepared with 0.5% CMC-Na solution and administered by gavage at a volume of 10 mL / kg one hour before LPS injection.

[0052] In drug preparation: Group G3: Accurately weigh 2.0g of TG powder, add 0.5% CMC-Na solution to 10mL, vortex to mix, and prepare a suspension with a concentration of 0.20g crude drug / mL; Group G4: Accurately weigh 2.0g of ICG powder, add 0.5% CMC-Na solution to 10mL, vortex to mix, and prepare a suspension with a concentration of 0.20g crude drug / mL; Group G5: Accurately weigh 2.0g of ICG-C powder, add 0.5% CMC-Na solution to 10mL, vortex to mix, and prepare a suspension with a concentration of 0.20g crude drug / mL; Group G6: Accurately weigh 4.0g of ICG-C powder, add 0.5% CMC-Na solution to 10mL, vortex to mix, and prepare a suspension with a concentration of 0.40g crude drug / mL; Group G2: Take 10 mL of pure water.

[0053] At the same dosage, the dosage for groups G3, G4, and G5 was 2.0 g crude drug / kg, with a gavage volume of 10 mL / kg, meaning a 200 g rat was given 2.0 mL of suspension by gavage; the dosage for group G6 was 4.0 g crude drug / kg, with a gavage volume of 10 mL / kg, meaning a 200 g rat was given 2.0 mL of suspension by gavage; and group G2 was given pure water by gavage, with a gavage volume of 10 mL / kg.

[0054] In Group G7, an appropriate amount of diazepam standard was accurately weighed, dissolved in sterile physiological saline, and prepared into a solution with a concentration of 0.05 mg / mL. The dosage was 0.5 mg / kg, and the intraperitoneal injection volume was 10 mL / kg.

[0055] Experimental steps: All animals were acclimatized for 3 days with free access to food and water. Before the experiment, the weight of each rat was measured and they were randomly divided into 7 groups according to their weight. The average weight of each group was recorded. Then, prophylactic drugs were administered 1 hour before LPS injection. Group G1 received no medication; Group G2 was administered pure water by gavage at a volume of 10 mL / kg. Group G3 was administered TG suspension by gavage (0.20 g crude drug / mL), volume 10 mL / kg, dosage 2.0 g crude drug / kg; Group G4 was administered ICG suspension by gavage (0.20 g crude drug / mL), volume 10 mL / kg, dosage 2.0 g crude drug / kg; Group G5 was administered ICG-C suspension (0.20 g crude drug / mL) by gavage, with a volume of 10 mL / kg and a dosage of 2.0 g crude drug / kg. Group G6 was administered ICG-C suspension (0.40 g crude drug / mL) by gavage, with a volume of 10 mL / kg and a dosage of 4.0 g crude drug / kg. Group G7 was administered pure water by gavage at a volume of 10 mL / kg. Pure water was administered to Group G7 as a placebo, while diazepam, the positive control drug, was injected intraperitoneally when the body was placed in a high-temperature incubator.

[0056] One hour after administration, rats in groups G2-G7 were intraperitoneally injected with lipopolysaccharide (LPS, Sigma L2880, 200 μg / kg, dissolved in sterile saline, injection volume 10 mL / kg). Rats in group G1 were intraperitoneally injected with an equal volume of sterile saline. Body temperature was then measured using a rectal thermometer before LPS injection and at 2, 4, and 6 hours after injection. Two and a half hours after LPS injection, rats in groups G2-G7 were individually placed in transparent acrylic observation boxes and placed in a constant temperature water bath oven at 40.0 ± 0.5℃ to induce seizures. Group G1 was placed in a room temperature environment without high temperature as a normal control. For Group G7, diazepam (0.5 mg / kg, injection volume 10 mL / kg) was injected intraperitoneally while being placed in the high-temperature chamber. From the start of placement in the high-temperature chamber, continuous video recording was performed for 30 minutes. The time until the first facial twitching or forelimb clonus was recorded as the seizure latency. The severity of seizures was recorded using the Racine grading system: Grade 1 was facial twitching; Grade 2 was head nodding; Grade 3 was forelimb clonus; Grade 4 was standing with forelimb clonus; and Grade 5 was standing and falling. The most severe grade for each animal during the observation period was recorded as the animal's score. Whether a seizure occurred was recorded, and the seizure incidence rate for each group was calculated. Group G1 was not placed in a high-temperature chamber and no seizures were observed; it was only used as a control for normal physiological indicators.

[0057] One-way ANOVA was used for comparisons among multiple groups. The LSD test was used for groups with homogeneous variances, and the Games-Howell test was used for pairwise comparisons of groups with unequal variances. The chi-square test was used to compare the incidence of seizures. P < 0.05 was considered statistically significant. Experimental results are as follows: Figure 2 , Figure 3 As shown in Tables 2 and 3.

[0058] Table 2 shows the severity scores of convulsions in each group of rats;

[0059] refer to Figure 2 In this experiment, the occurrence of seizures in rats of each group during the observation period was recorded simultaneously, and the seizure incidence rate was calculated. The relevant data are summarized in Table 3: Table 3 shows the comparison of the main pharmacodynamic indicators of febrile seizures in each group of rats.

[0060] The results showed that, under the same dosage, the seizure latency in group G5 was 785±124 seconds, which was significantly longer than that in group G3 (521±98 seconds) and group G4 (598±112 seconds) (P<0.05).

[0061] Meanwhile, the incidence of seizures in group G5 decreased to 45%, lower than 85% in group G3 and 75% in group G4; its mean seizure severity score (1.8±0.8) was also significantly lower than that in group G3 (3.3±0.9) and group G4 (2.9±1.0) (P<0.05). Targeted particles can significantly reduce seizure severity in a dose-dependent manner.

[0062] Experiment 3: Validation of Gut Microbiota Regulation Samples: Colon contents of rats in groups G2, G3, G5, and G6 of Experiment 2 (n=2 / group); After quality control and noise reduction, the raw data were used to generate ASV (Amplicon Sequence Variant) tables. Species were annotated based on the Silva138 database. Alpha diversity was analyzed using the Shannon index and the ObservedASVs index. Beta diversity was analyzed using principal coordinate analysis (PCoA) based on Bray-Curtis distance. The PERMANOVA test was used to test differences between groups. The LEfSe (LDA>3.5) was used to identify marker species for differences between groups. The Shannon index of group G6 was significantly higher than that of groups G2 and G3, indicating that it could significantly restore the decline in microbial diversity caused by the model.

[0063] PCoA analysis showed that the sample points of groups G2, G3 and G5, G6 were clearly separated along the PC1 axis (PERMANOVA, P<0.01), indicating that ICG-C intervention significantly altered the overall microbial community structure.

[0064] refer to Figure 4 This experiment analyzed the microbiota in the colon contents of rats in each group, focusing on the changes in the relative abundance of beneficial bacteria genera. Relevant data are shown in Table 4. Table 4 shows the relative abundance changes of key beneficial bacteria genera;

[0065] In group G6, Lactobacillus and Bifidobacterium were significantly identified as key biomarkers (LDA>4.0).

[0066] Comparative Example 1 This comparative example uses a conventional process for combining and extracting all components of traditional Chinese medicine to prepare granules. It is used to compare the differences in safety and efficacy between the technical solution of this invention and the traditional process, and to verify the synergistic effect of the combination of technologies such as detoxification of processing, segmented extraction, addition of prebiotics, and double-layer coating in this invention.

[0067] In the preparation method, raw Sichuan pepper, raw sophora flavescens, raw costus root, raspberry wood, galangal, chebula, gardenia, uncaria, and cicada slough are weighed out. All medicinal materials are not processed in any way. All the above medicinal materials are placed into a multi-functional extraction tank at once, and 10 times the amount of drinking water is added. The mixture is heated to boiling and then maintained at a gentle boil for 2.5 hours to complete the first extraction. After filtering the liquid, 8 times the amount of drinking water is added to the residue, and a second extraction is performed under the same conditions for 1.5 hours. The two extracts are combined, filtered through a 200-mesh filter cloth, and concentrated under reduced pressure at 65-70℃ and -0.08MPa until a thick paste with a relative density of 1.28 at 60℃ is obtained. This thick paste is then mixed evenly with the prescribed amounts of rock sugar powder and white sugar powder in a mixer. An appropriate amount of 70% ethanol is added as a wetting agent to form a soft mass, which is then granulated using a gyratory granulator through a 14-mesh sieve. The obtained wet granules were dried in a 55℃ hot air circulating drying oven until the moisture content was ≤5.0%, and finally granulated through an 80-mesh sieve to obtain uncoated traditional Erdun-7 granules (TG).

[0068] This comparative study does not involve any processing or detoxification of the medicinal materials. All medicinal materials are extracted together in one go. No prebiotics such as Cynomorium songaricum polysaccharide or Hippophae rhamnoides oligosaccharide are added, and no double-layer coating is used for targeted delivery.

[0069] To verify the differences between the traditional process and the process of this invention, the following tests were conducted: The content of azadirachtin was determined by high performance liquid chromatography, and the results are as follows; Table 5 compares the azadirachtin content between Comparative Example 1 and Example 1:

[0070] The results showed that, in Example 1, the azadirachtin content was reduced compared to that in Comparative Example 1 by processing Sichuan chinaberry with vinegar, which significantly reduced the potential risk of hepatotoxicity.

[0071] Next, 20 SPF-grade SD rats were randomly divided into two groups, and administered granules of Comparative Example 1 and Example 1 respectively by gavage for 7 consecutive days. After the last administration, the rats were dissected to observe the pathological changes in the gastric mucosa and scored according to the damage scoring criteria. Table 6 shows a comparison of gastric mucosal injury scores between Comparative Example 1 and Example 1:

[0072] The results showed that, through processes such as charring and double-layer coating, the gastric mucosa irritation of Example 1 was significantly reduced, and its safety was significantly better than that of traditional processes.

[0073] Referring to the pharmacodynamic experimental method in Example 1, the anticonvulsant effects of Comparative Example 1 and Example 1 were compared, and the key indicators are as follows; Table 7 compares the anticonvulsant effects of Comparative Example 1 and Example 1:

[0074] The results showed that the targeted Erdun-7 particles of Example 1 were significantly superior to the conventional Erdun-7 particles (TG) prepared by the conventional process in terms of key indicators such as seizure latency, incidence, and severity.

[0075] Comparative Example 2 This comparative study, while retaining the addition of prebiotics and double-layer coating, omits specific detoxification treatments for the medicinal materials, and is used to verify the necessity of adding detoxification treatments to vinegar-processed Sichuan chinaberry, charred sophora flavescens, and coarse powder of costus root.

[0076] Weigh out the prescribed amounts of raw Sichuan pepper and raw sophora flavescens, without processing them with vinegar or charcoal. Also, leave the costus root unprocessed. Combine all the herbs and place them in an extraction tank. Add 10 times the amount of drinking water for the first decoction, which lasts 1.5 hours. After filtering the liquid, add 8 times the amount of water to the dregs for a second decoction, which lasts 1.0 hour. Combine the filtrates and filter again. Concentrate the mixture to a thick paste with the same relative density as in Example 1. Then, mix this paste with the prescribed amounts of Cynomorium songaricum polysaccharide extract, Hippophae rhamnoides oligosaccharide extract, rock sugar powder, and white sugar powder. Prepare uncoated granules using the same wet granulation parameters as in Example 1. Finally, completely replicate the coating process of Example 1 for these granules. In a fluidized bed coating machine, alternately spray with 1.0% chitosan solution and 1.0% sodium alginate solution to form a protective layer, then spray with 2.0% cholic acid-PLGA solution to form a targeting layer. After curing and drying, obtain comparative granules with targeting function.

[0077] In this comparative example, raw Sichuan chinaberry and raw sophora flavescens are not processed, thus retaining their original toxic components; costus root is not coarsely ground or post-processed, and is decocted throughout the entire process; the addition of prebiotics and double-layer coating are the same as in Example 1.

[0078] To verify the detoxification process of the preparation method, the contents of azadirachtin and matrine were first determined by high performance liquid chromatography. The results are as follows. Table 8 compares the content of toxic components between Comparative Example 2 and Example 1:

[0079] Referring to the pharmacodynamic experimental methods in Example 1, the anticonvulsant effects of Comparative Example 2 and Example 1 were compared, and the key indicators are as follows; Table 9 shows the comparison of the anticonvulsant effects between Comparative Example 2 and Example 1:

[0080] The results showed that although Comparative Example 2 retained the double-layer coating and prebiotic addition process, its anticonvulsant effect was still significantly lower than that of Example 1 due to the lack of processing to reduce toxicity. This indicates that processing to reduce toxicity not only improves safety but also enhances overall efficacy by optimizing the ratio of active ingredients.

[0081] The technical scope of this invention is not limited to the content described above. Those skilled in the art can make various modifications and variations to the above embodiments without departing from the technical concept of this invention, and all such modifications and variations should fall within the protection scope of this invention.

Claims

1. An Erdun-7 granule preparation, characterized in that, The ingredients include the following parts by weight: One part of Aucklandia lappa and 1.3-1.8 parts of Rubus idaeus are used to regulate Qi, relieve exterior symptoms, and reduce fever. Sophora flavescens 1.6-2.4 parts, Gardenia jasminoides 0.7-1.3 parts, Terminalia chebula 0.7-1.3 parts are used to enhance the heat-clearing, fire-purging, detoxifying, and blood-cooling effects of Aucklandia lappa and Rubus idaeus. Uncaria rhynchophylla 0.8-1.2 parts and Cicadae periostracum 0.8-1.2 parts are used to calm the liver and extinguish wind, calm fright and stop spasms; Kaempferia galanga 0.7-1.3 parts and Melia toosendan 0.7-1.3 parts are used to promote qi circulation and relieve pain, soothe the liver and clear heat, regulate qi and blood to help the medicine exert its effects. Cynomorium songaricum 0.8-1.2 parts and Hippophae rhamnoides 0.8-1.2 parts are used to invigorate the spleen and replenish essence. Cynomorium songaricum is used to nourish the blood and replenish essence. 3.0-6.0 parts rock sugar, 3.0-6.0 parts white sugar; The Erdon-7 granules are coated with a double layer consisting of a protective layer and a targeting layer. The protective layer is composed of chitosan and sodium alginate, which maintains structural integrity in the acidic environment of the stomach and dissolves in the alkaline environment of the intestine. The targeting layer is a bile acid-modified polylactic acid-glycolic acid copolymer, which enables the active delivery of the Erdon-7 granules to the target area of ​​the intestine through the specific binding of bile acids to bile acid receptors in the intestine.

2. The Erdun-7 granule medicine according to claim 1, characterized in that: The Cynomorium songaricum and the Hippophae rhamnoides are added to the Erdun-7 granule medicine in the form of extracts, and the Cynomorium songaricum polysaccharide in Cynomorium songaricum and the Hippophae rhamnoides oligosaccharide in Hippophae rhamnoides are used to regulate the intestinal flora.

3. A method for preparing Erdun-7 granules according to any one of claims 1-2, characterized in that: Includes the following steps: Step 1: Provide raw materials: The raw materials include costus root, sophora flavescens, gardenia, chebula, uncaria, cicada slough, galangal, chinaberry, cynomorium, sea buckthorn, rock sugar, and white sugar; First, take Sichuan chinaberry and process it with vinegar; take sophora flavescens and process it with charcoal; take costus root and grind it into coarse powder for later use; then mix the remaining medicinal materials, including gardenia, chebula, uncaria, cicada slough, and galangal. Step 2: Mix the processed products obtained from Sichuan chinaberry and sophora flavescens in Step 1 with the other medicinal materials except for costus root, and carry out the first round of extraction; Step 3: Next, combine the remaining medicinal materials from Step 1 with the coarse powder of *Inula japonica* and carry out the second round of extraction; the coarse powder of *Inula japonica* should be added 30-60 minutes before the end of the second round of extraction. Step 4: Extracting Cynomorium songaricum and Hippophae rhamnoides to obtain the first extract, Cynomorium songaricum polysaccharide and Hippophae rhamnoides oligosaccharide; Step 5: Concentrate the extract from Step 2 into a thick paste, mix it with Cynomorium songaricum polysaccharide, Hippophae rhamnoides oligosaccharide, rock sugar and white sugar and granulate to obtain uncoated Erdun-7 granules. Step Six: Sequentially coat the uncoated Erdun-7 granules with a protective layer and a targeting layer. The protective layer is formed by alternately spraying chitosan solution and sodium alginate solution and then ionically crosslinking them. The targeting layer is formed by spraying a cholic acid-polylactic acid-glycolic acid copolymer solution.

4. The method for preparing Erdun-7 granules according to claim 3, characterized in that: For every 100 parts by weight of Sichuan chinaberry, mix with 20-30 parts by weight of rice vinegar, let it soak for 1-2 hours, then stir-fry over low heat at 80-120℃ for 15-30 minutes until the surface of the herb darkens and the vinegar smell dissipates.

5. The method for preparing Erdun-7 granules according to claim 3, characterized in that: The process of charring involves placing the Sophora flavescens slices in a frying pan at 180-220℃ and stir-frying them rapidly over high heat for 5-10 minutes until the surface of the herb is charred black and the inside is charred brown. The weight loss rate of the herb is controlled within the range of 15%-25%. Then, 10%-15% of the weight of the herb is sprayed with water to extinguish the smoke, and the herb is taken out and spread out to dry.

6. The method for preparing Erdun-7 granules according to claim 3, characterized in that: The extraction time for the first round in step two is 1.5-2.5 hours, and the extraction time for the second round in step three is 0.5-1.5 hours; wherein the extraction time for the first round is at least 0.5 hours longer than the extraction time for the second round.

7. The method for preparing Erdun-7 granules according to claim 3, characterized in that: When alternately spraying the chitosan solution and sodium alginate solution, first spray the chitosan solution, and after drying, spray the sodium alginate solution.

8. The method for preparing Erdun-7 granules according to claim 3, characterized in that: The coating weight gain of the protective layer is 3%-5%, and the coating weight gain of the targeting layer is 5%-8%.

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