Traditional Chinese medicine composition for treating infantile recurrent respiratory tract infection

Abstract

The invention relates to a traditional Chinese medicine composition for treating infantile recurrent respiratory tract infection, and relates to the technical field of traditional Chinese medicines, the traditional Chinese medicine composition comprises 6 g of radix pseudostellariae, 6 g of roasted rhizoma atractylodis macrocephalae, 10 g of poria cocos, 3 g of raw liquorice, 10 g of raw radix astragali, 6 g of radix saposhnikoviae, 6 g of honeysuckle flower, 6 g of mint, 6 g of rhizoma pinellinae praeparata, 6 g of pericarpium citri reticulatae, 6 g of platycodon grandiflorum, 10 g of radix peucedani, 15 g of aster, 10 g of radix stemonae, 6 g of exocarpium citri grandis The core principle of the traditional Chinese medicine composition for treating infantile recurrent respiratory tract infection is to strengthen the body resistance to eliminate pathogenic factors, treat both symptoms and root causes, and improve the pathological basis of recurrent infection by virtue of the multi-layer effects of tonifying lung and spleen, consolidating superficies and tonifying qi, clearing heat and eliminating phlegm, and ventilating lung and relieving cough according to the physique characteristics of'common insufficiency of lung and spleen 'and'non-consolidation of external defensive qi' of children.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicines, and in particular to a traditional Chinese medicine composition for treating recurrent respiratory tract infections in children. Background Art

[0002] Recurrent respiratory tract infection in children is one of the common diseases in pediatric clinical practice. When the number of upper and lower respiratory tract infections in children increases and exceeds a certain range, it is called recurrent respiratory tract infection, or recurrent infection for short. Traditional Chinese medicine believes that the delicate constitution of children and the imperfect functions of the lungs, spleen and kidneys are important internal factors leading to recurrent respiratory tract infections. Specifically: Lung defense is not solid: The lungs control the skin and hair, respiration, and the exterior and interior. If the lung qi is weak, it cannot play its role in defending against external evil spirits normally, making it easy for external evil spirits such as wind, cold, dampness and heat to invade the human body; spleen and stomach weakness: The spleen and stomach are the foundation of acquired constitution, responsible for transporting and transforming the essence of water and grain and supplying nutrition to the whole body. If the spleen and stomach function poorly, the source of qi and blood will be insufficient, the body's resistance will be reduced, and infection will be prone to occur; congenital deficiency or insufficient kidney qi: The kidneys store essence, control growth, development and reproduction, and are the foundation of human life activities. Children's kidney qi is not yet sufficient, especially premature babies and low birth weight babies, who are more prone to recurrent respiratory tract infections. The causes of recurrent respiratory tract infections in children are different and can be caused by either deficiency or excess, so they need to be treated fundamentally.

[0003] For example, Chinese patent application number CN201810708175.6 discloses a traditional Chinese medicine composition for treating upper respiratory tract diseases. Its purpose is to provide a traditional Chinese medicine composition for treating recurrent respiratory tract infections in children that has the effects of regulating immunity, controlling inflammation, and protecting against infection. The traditional Chinese medicine composition for treating recurrent respiratory tract infections in children comprises the following ingredients in parts by weight: 5-15 grams of Astragalus membranaceus, 5-15 grams of Saposhnikovia divaricata, 5-15 grams of Angelica dahurica, 5-15 grams of Xanthium sibiricum fruit, 5-15 grams of Magnolia flower, 5-15 grams of Apricot kernel, 5-15 grams of stir-fried Tripterygium wilfordii fruit, and 5-15 grams of raw liquorice. This invention is based on the Traditional Chinese Medicine theory that the lungs and large intestine are exterior and interior components of each other. The formulation of this medicine features the regulation of the lungs and large intestine. This invention has the effects of regulating immunity, controlling inflammation, and protecting against infection.

[0004] However, the causes of recurrent respiratory tract infections in children are complex and the treatment cycle is long. Long-term use of laxatives such as almonds and lettuce seeds may aggravate spleen and stomach weakness in children, leading to diarrhea or indigestion and worsening of the condition. In addition, it is only designed around the theory of "the lungs and large intestine are exterior and interior", and does not cover common syndromes such as phlegm heat and wind heat. It may have limited effect on complex cases (such as those with high fever and yellow sputum). Summary of the Invention

[0005] The present invention aims to solve the technical problems existing in the prior art and provides a traditional Chinese medicine composition for treating recurrent respiratory tract infections in children.

[0006] The technical solution of the present invention for solving the above technical problems is as follows: a traditional Chinese medicine composition for treating recurrent respiratory tract infections in children, comprising 6g of Pseudostellariae Radix, 6g of stir-fried Atractylodes macrocephala, 10g of Poria cocos, 3g of raw licorice, 10g of raw Astragalus membranaceus, 6g of Saposhnikovia divaricata, 6g of honeysuckle, 6g of peppermint, 6g of Amomum villosum, 6g of Citrus reticulata, 6g of Platycodon grandiflorum, 10g of Peucedanum peucedanum, 15g of Aster scutellariae, 10g of Stemona tuber, 6g of Citrus aurantium, and 20g of Houttuynia cordata.

[0007] Furthermore, the preparation method is: S1. Wash the formula and dry it at 60°C, crush it and sieve it with 60 mesh, then place the Chinese medicine powder in a supercritical extraction device for extraction, add entrainer to the entrainer tank, the mass ratio of powder to entrainer is 20:3, soak it for 30 minutes, and then perform supercritical carbon dioxide extraction; S2. The mixed solution was placed in a vacuum distillation apparatus, 1.2 parts of a stabilizer 6-chloronicotinic acid was added, and the mixture was stirred for 15 minutes. The pressure was set to 15 mmHg and the temperature was set to 70°C. After reacting for 2 hours, 0.4 parts of a stabilizer 6-chloronicotinic acid was added and the mixture was reacted for another 2 hours to obtain a pure extract. S3. Take 100 parts of pure extract and add it to the drum equipment. Add 3 parts of maltose, 0.5 parts of citric acid, and 5 parts of sodium chloride. The temperature is 35°C. Add 5 parts of the auxiliary agent FMOC-L-phenylalanine to the drum equipment every two hours. The drum speed is 100 r / min. React for a total of 5 hours and finally flow out to obtain the product.

[0008] Furthermore, the CO2 flow rate in the supercritical carbon dioxide extraction was 25 L / h, and the extraction was divided into three stages. The first stage had a pressure of 15 MPa, an extraction temperature of 40 °C, and a time of 20 min; The second stage pressure is 25 MPa, the extraction temperature is 35°C, and the extraction time is 30 min; The pressure of the third stage is 35 MPa, the extraction temperature is 32°C, and the extraction time is 40 min. After the extraction is completed, the material is filtered and the mixed liquid is collected.

[0009] Furthermore, the entrainer includes, by mass, 0.8 parts of a hydrophobic group, 0.5 parts of a wrapping group, 0.2 parts of a separation group, and 6.0 parts of a solvent.

[0010] Furthermore, the hydrophobic group is prepared by adding 0.1% lipase CAL-B to a mixture of L-histidine and Boc anhydride in a mass ratio of 1:0.25, reacting at 35 MPa / 35°C for 30 min, adding 3Å molecular sieves to adsorb the generated tert-butanol and water, and decompressing to 5 MPa. CO2 is discharged with the unreacted Boc anhydride, and the product Boc-histidine is retained in the reactor.

[0011] Furthermore, the encapsulation group was mixed with polyethylene glycol and polycaprolactone in a mass ratio of 4:1, and stannous octoate was added as a catalyst to react for 6 hours under a nitrogen environment at 140°C to generate a PEG-PCL block copolymer.

[0012] Furthermore, the isolated group is Pluronic L31, which is a PEO-PPO-PEO triblock copolymer with a molecular weight of 1100 Da.

[0013] Furthermore, Fe3O4@SiO2-CAL-B is additionally added to the hydrophobic group, and the Fe3O4@SiO2-CAL-B in the hydrophobic group is 0.5 parts by weight. The specific preparation method is as follows: FeCl2·4H2O and FeCl3·6H2O were dissolved in deionized water at a molar ratio of 1:2 and stirred under nitrogen. NH3·H2O was then added dropwise until the pH reached 11. The mixture was reacted at 80°C for 1 hour to generate Fe3O4 precipitate. The mixture was then washed with magnetic separation until neutral and dried in vacuum to obtain Fe3O4 nanoparticles. Fe3O4 was dispersed in an ethanol-water (4:1) solution, and TEOS (tetraethyl orthosilicate) and NH3·H2O were added. The mixture was reacted at 40°C for 6 h to form a SiO2 shell. The mixture was then magnetically separated, washed, and dried to obtain Fe3O4@SiO2. Fe3O4@SiO2 was dispersed in toluene, APTES was added, and the mixture was reacted at 80°C for 12 h to introduce amino groups. Glutaraldehyde was added to react with the amino groups of lipase CAL-B. The mixture was fixed at 25°C for 6 h, and free enzyme was removed by magnetic separation to obtain Fe3O4@SiO2-CAL-B.

[0014] Furthermore, instantaneous decompression is also performed during the carbon dioxide extraction, specifically: In the three stages of extraction, the first stage pressure was 15 MPa, the extraction temperature was 40°C, and the extraction time was 20 min; after the first stage, the pressure was pulsed down to 5 MPa; The pressure in the second stage is 25 MPa, the extraction temperature is 35°C, and the extraction time is 30 min. During the second stage, the pressure is instantly reduced to 3 MPa and then restored every 5 min. The pressure in the third stage is 35 MPa, the extraction temperature is 32°C, and the extraction time is 40 min. After the third stage, the pressure is instantly reduced to 10 MPa. After the extraction is completed, the material is filtered and the mixed liquid is collected.

[0015] Furthermore, pretreatment was performed before the first stage of carbon dioxide extraction. The product was crushed to 100 mesh at -20 °C and soaked in 5 MPa liquid CO2 for 30 min with ultrasound assistance. Then, the product was cycled from 5 MPa to 15 MPa for 3 times, with a total time of 15 min. The pressure was then increased stepwise from 5 MPa to the pressure required for the first stage.

[0016] The beneficial effects of the present invention are: First, the core principle of this Chinese medicine composition in treating recurrent respiratory tract infections in children lies in strengthening the body's resistance and eliminating evil, treating both the symptoms and the root causes. Through the multi-level effects of nourishing the lungs and spleen, consolidating the exterior and invigorating qi, clearing away heat and resolving phlegm, and promoting lung function and relieving cough, it targets the physical characteristics of children's "lung and spleen are often insufficient" and "the defense of the body is not solid", and improves the pathological basis of recurrent infections.

[0017] Secondly, the design of the entrainer embodies a multi-level molecular engineering strategy. The entrainer system improves the total flavonoid extraction rate and greatly retains the heat-sensitive component menthone through multi-scale synergy of molecular recognition (Boc group), nano-confinement (micelles), interface regulation (Pluronic) and solvent polarity.

[0018] Third, the Boc protection effect was enhanced by Fe3O4@SiO2-CAL-B, and the CO2 penetration depth was increased through pretreatment. DETAILED DESCRIPTION

[0019] The following is a clear and complete description of the technical solutions in the embodiments of this application. Obviously, the embodiments described are only part of the embodiments of this application, not all of them. Based on the embodiments in this application, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of this application.

[0020] In the description of this application, the terms "first" and "second" are used for descriptive purposes only and should not be understood to indicate or imply relative importance or implicitly specify the number of the technical features indicated. Therefore, a feature specified as "first" or "second" may explicitly or implicitly include one or more of the described features. In the description of this application, "plurality" means two or more, unless otherwise specifically specified.

[0021] In the description of this application, the term "for example" is used to mean "used as an example, illustration or explanation". Any embodiment described in this application as "for example" is not necessarily to be construed as being more preferred or advantageous than other embodiments. The following description is given to enable any person skilled in the art to implement and use the present invention. In the following description, details are listed for the purpose of explanation. It should be understood that a person of ordinary skill in the art can recognize that the present invention can be implemented without using these specific details. In other examples, well-known structures and processes are not elaborated in detail to avoid obscuring the description of the present invention with unnecessary details. Therefore, the present invention is not intended to be limited to the embodiments shown, but is consistent with the widest scope consistent with the principles and features disclosed in this application.

[0022] Example 1: A traditional Chinese medicine composition for treating recurrent respiratory tract infections in children, comprising 6g of Pseudostellariae Radix, 6g of stir-fried Atractylodes macrocephala, 10g of Poria cocos, 3g of raw licorice, 10g of raw Astragalus, 6g of Saposhnikovia divaricata, 6g of honeysuckle, 6g of peppermint, 6g of Amomum villosum, 6g of Citrus reticulata, 6g of Platycodon grandiflorum, 10g of Peucedanum peucedanum, 15g of Aster scutellariae, 10g of Stemona tuber, 6g of Citrus aurantium, and 20g of Houttuynia cordata.

[0023] The preparation method of the above-mentioned Chinese medicine composition is: S1. Wash the formula and dry it at 60°C, crush it and sieve it with 60 mesh, then place the Chinese medicine powder in a supercritical extraction device for extraction, add entrainer to the entrainer tank, the mass ratio of powder to entrainer is 20:3, soak it for 30 minutes, and then perform supercritical carbon dioxide extraction; The CO2 flow rate during extraction was 25 L / h, and the extraction was divided into three stages. The first stage had a pressure of 15 MPa, an extraction temperature of 40 °C, and a time of 20 min. The second stage pressure is 25 MPa, the extraction temperature is 35°C, and the extraction time is 30 min; The third stage pressure is 35 MPa, the extraction temperature is 32°C, and the extraction time is 40 min. After the extraction is completed, the material is filtered and the mixed liquid is collected; S2. The mixed solution was placed in a vacuum distillation apparatus, 1.2 parts of a stabilizer 6-chloronicotinic acid was added, and the mixture was stirred for 15 minutes. The pressure was set to 15 mmHg and the temperature was set to 70°C. After reacting for 2 hours, 0.4 parts of a stabilizer 6-chloronicotinic acid was added and the mixture was reacted for another 2 hours to obtain a pure extract. S3. Take 100 parts of pure extract and add it to the drum equipment. Add 3 parts of maltose, 0.5 parts of citric acid, and 5 parts of sodium chloride. The temperature is 35°C. Add 5 parts of the auxiliary agent FMOC-L-phenylalanine to the drum equipment every two hours. The drum speed is 100 r / min. React for a total of 5 hours and finally flow out to obtain the product.

[0024] Wherein, by mass, the entrainer includes 0.8 parts of a hydrophobic group, 0.5 parts of a wrapping group, 0.2 parts of a separation group, and 6.0 parts of a solvent; The hydrophobic group is prepared by mixing L-histidine and Boc anhydride at a mass ratio of 1:0.25, adding 0.1% (w / w) lipase CAL-B (Novozym 435), and reacting at 35 MPa / 35°C for 30 min. 3Å molecular sieves (10% w / w) are added to adsorb the generated tert-butanol and water. The pressure is then reduced to 5 MPa, and the unreacted Boc anhydride is discharged with CO2, while the product Boc-histidine remains in the reactor. The encapsulation group was prepared by mixing polyethylene glycol (PEG500) and polycaprolactone (PCL2000) in a mass ratio of 4:1. Stannous octoate (0.1% w / w) was added as a catalyst and reacted for 6 h at 140°C in a nitrogen environment to generate a PEG-PCL block copolymer. The isolated group is Pluronic L31, which is a PEO-PPO-PEO triblock copolymer (PEO accounts for 10%, PPO accounts for 90%), with a molecular weight of 1100 Da; The solvent is a water solvent of propylene glycol, which is obtained by evenly mixing propylene glycol and water in a volume ratio of 5:2.

[0025] The technical solutions in the above embodiments of the present application have at least the following technical effects or advantages: The core principle of this Chinese medicine composition in treating recurrent respiratory tract infections in children is to strengthen the body's resistance and eliminate evil, and treat both the symptoms and the root causes. Through the multi-level effects of nourishing the lungs and spleen, consolidating the exterior and invigorating qi, clearing away heat and resolving phlegm, and promoting lung function and relieving cough, it targets the physical characteristics of children's "often insufficient lungs and spleen" and "weak defense against external diseases", and improves the pathological basis of recurrent infections.

[0026] Prince ginseng and astragalus: invigorate qi and strengthen the spleen, nourish the lungs and consolidate the exterior, enhance the ability to defend against external pathogens; stir-fried white atractylodes, poria, and licorice: a modification of Sijunzi Decoction, invigorate the spleen and remove dampness to nourish the acquired foundation, and assist in the production of qi and blood; Yupingfeng Powder (astragalus + white atractylodes + saposhnikovia): consolidates the exterior and stops sweating, reduces the invasion of external pathogens, removes pathogens and treats the symptoms (clears heat and resolves phlegm, promotes lung function and relieves cough); honeysuckle and houttuynia: clears heat and detoxifies, targeting phlegm-heat stagnation in the lungs common in respiratory tract infections; peucedanum, aster, radix stemonae, and platycodon: promotes and descends lung qi, relieves cough and resolves phlegm, and regulates stagnation of lung qi; fennel, tangerine peel, and red orange peel: regulate qi, dry dampness, and resolve phlegm, and resolve phlegm stagnation in the lungs; peppermint: dispels wind and dissipates heat, relieves sore throat, and penetrates the exterior pathogens.

[0027] In the Qi-tonifying and exterior-strengthening group, Pseudostellaria baicalensis, Astragalus membranaceus, and Atractylodes macrocephala replenish the Qi of the lungs and spleen and enhance immunity; Astragalus membranaceus is combined with Atractylodes macrocephala and Saposhnikovia divaricata to form Yupingfeng Powder, which strengthens the defense function of the exterior defense; in the Spleen-strengthening and Damp-removing group, Poria cocos, Licorice root, and Tangerine peel strengthen the spleen and eliminate dampness, eliminating the source of phlegm; Tangerine peel regulates Qi and harmonizes the middle and helps the spleen transport and transform; in the Heat-clearing and phlegm-resolving group, Honeysuckle, Houttuynia cordata, and Huatangerine peel Honeysuckle penetrates the surface heat, Houttuynia cordata clears lung heat, and Tangerine peel resolves phlegm, and together they resolve phlegm-heat that blocks the lungs; in the Lung-promoting and Cough-relieving group, Platycodon grandiflorum, Peucedanum chinense, Aster strychnine, and Stemona bulb Platycodon grandiflorum promotes the lungs (ascending), Peucedanum chinense descends Qi (descending), one ascending and one descending to regulate lung Qi; Aster strychnine and Stemona bulb moisten the lungs and relieve cough; in the Wind-dispersing and Evil-removing group, Peppermint, Saposhnikovia divaricata and Peppermint dispel wind and dissipate heat, while Saposhnikovia divaricata dispels wind and resolves the exterior, working together to disperse external evil; in the Qi-regulating and phlegm-resolving group, Summerleaf truncatum, Summerleaf truncatum, and Huatangerine peel Summerleaf dries dampness and resolves phlegm, while Summerleaf

[0028] Children's lungs and spleens are often weak, so the tonic herbs in the formula are light and gentle (such as replacing ginseng with pseudoginseng) to avoid congestion; heat-clearing herbs (honeysuckle, houttuynia cordata) are combined with lung-clearing herbs (mint, platycodon grandiflorum) to ensure that the pathogens are eliminated without harming the body. Yupingfeng Powder can regulate the Th1 / Th2 immune balance and reduce the recurrence of infection; Houttuynia cordata contains ingredients such as quercetin, which inhibit respiratory pathogens; Aster ternata and Stemona radix can relieve airway hyperresponsiveness and reduce persistent cough.

[0029] Through the three-dimensional combination of strengthening the spleen and resolving phlegm (Atractylodes macrocephala, Poria cocos), clearing away heat (Lonicera japonica, Houttuynia cordata), and regulating Qi (Cited Tangerine Peel, Platycodon grandiflorum), phlegm-Qi-heat are treated simultaneously, blocking the vicious cycle of "spleen deficiency causing phlegm → phlegm stagnation turning into heat → lung Qi stagnation → repeated infection".

[0030] This prescription treats both the symptoms and the root cause by tonifying the lungs and spleen to strengthen the body, clearing phlegm and heat to dispel pathogenic factors, and promoting the descending of lung qi to relieve cough. The drug combination emphasizes both lightness, clarity, and penetration, combining attack and nourishment. This approach aligns with the pathological characteristics of children, who are prone to both deficiency and excess. It can improve the underlying constitution of recurrent infections, rapidly alleviate acute symptoms, and reduce recurrences.

[0031] The design of the entrainer embodies a multi-level molecular engineering strategy. The entrainer system improves the total flavonoid extraction rate and greatly retains the heat-sensitive component menthone through multi-scale synergy of molecular recognition (Boc group), nanoconfinement (micelles), interface regulation (Pluronic) and solvent polarity.

[0032] Boc anhydride reacts with the imidazole ring (C3H4N2) of L-histidine through a nucleophilic substitution reaction to form an N-Boc protected structure, creating a steric hindrance. The tert-butyloxycarbonyl group of the Boc group (C5H9O2) is highly hydrophobic (LogP = 1.8), forming hydrophobic-hydrophobic interactions with fat-soluble components of traditional Chinese medicine (such as astragaloside IV). In supercritical CO2 (dielectric constant ε ≈ 1.5), Boc-histidine forms a molecular bridge through its amphiphilic structure: the NH of the imidazole ring forms hydrogen bonds (bond energy ≈ 5 kcal / mol) with water-soluble components (such as chlorogenic acid from honeysuckle), and the Boc group forms van der Waals interactions with CO2 (bond energy ≈ 0.5 kcal / mol). The Flory-Huggins interaction parameter χ = 0.12 (140°C) between PEG500 (EO chain) and PCL2000 (CL chain) drives the formation of core-shell micelles (Dh ≈ 25 nm). The PCL core forms π-π stacking (spacing ≈ 3.4 Å) with Houttuynia cordata flavonoids via ester groups (-OCO-). Under supercritical conditions of 35 MPa, CO2 swelling increases the micelle porosity by 40%, boosting the encapsulation efficiency of volatile components such as menthol (LogP = 3.3) to 78%. The PPO chain segment (LogS = 2.1) contracts conformationally in high-pressure CO2, forming a surface tension gradient. The PEO end (HLB = 3) is anchored at the CO2 / water interface, reducing the interfacial energy. Phase diagram analysis showed that Pluronic L31 increased the cloud point temperature of the extraction system to 45°C, promoting a 35% increase in the molecular sieving efficiency of aster saponins (molecular weight 842 Da) and impurity polysaccharides (molecular weight > 5kDa).

[0033] The polarity gradient of the propylene glycol / water (5:2) mixed solvent was formed. Through the reorganization of the hydrogen bond network, the hydroxyl group (-OH) of propylene glycol and CO2 formed a carbonate transition state, which promoted the dissolution of praeruptorin (polarity 0.32). Water molecules were transferred through the Grotthuss proton chain mechanism to maintain the stable dispersion of Pseudostigma polysaccharide. Under the three-stage pressure gradient (15→35 MPa), the CO2 density increased from 0.65 g / cm 3 Increased to 0.95 g / cm³, and the dielectric constant increased from 1.3 to 1.6; Stage 1 (15MPa): Low-density CO2 selectively extracts small-molecule terpenes (such as tangerine peel limonene); Stage 2 (25MPa): Medium-density fluid dissolves medium-polarity components (platycodonoside D, LogP=2.1); Stage 3 (35MPa): High-density supercritical state destroys the lignin-cellulose network of plant cell walls (the activation energy of hydrogen bond breaking is reduced by 23%), releasing bound components (stemona alkaloids).

[0034] The effect of the Chinese herbal medicine composition on treating recurrent respiratory tract infections in children was verified through experiments, and experiments were designed to verify the preparation and extraction effects; The efficacy of the Chinese herbal medicine composition in treating recurrent respiratory tract infections in children was verified by animal experiments. Cyclophosphamide and pathogenic bacteria were used to establish a model. Specifically, 3-4 week-old SD rats weighing 80-100 g were selected. After one week of adaptive culture, cyclophosphamide (75 mg / kg rat body weight) was injected intraperitoneally for 3 consecutive days. Then, a suspension of Streptococcus pneumoniae (10 7CFU / mL), and observed for 7 days after infection. The model was successfully established when the body weight decreased by more than 15%. Then the group experiment was carried out, including 10 normal control groups (nebulized normal saline), 10 model control groups (no treatment after infection), 20 positive drug groups (daily oral administration of azithromycin 10 mg / kg rat body weight), 20 low-dose Chinese medicine groups (daily oral administration of extract 0.5 g / kg rat body weight), 20 medium-dose Chinese medicine groups (daily oral administration of extract 1.0 g / kg rat body weight), and 20 high-dose Chinese medicine groups (daily oral administration of extract 2.0 g / kg rat body weight). On the 7th and 14th days, rat-specific IL-6 and TNF-α were used to treat the supernatant of lung tissue homogenate. Inflammatory factors were detected by ELISA kits. Peripheral blood was anticoagulated with heparin and lymphocytes were extracted by gradient centrifugation of lymphocyte separation fluid (Ficoll). Flow cytometry was used to detect immune indicators (CD4+ / CD8+ ratio). The results on day 7 are shown in Table 1, and the results on day 14 are shown in Table 2. The weight recovery rate is: (current weight - lowest weight after modeling) / basal weight × 100%. Significance mark: compared with the model control group, p<0.05, p<0.01; Table 1 One week after medication

[0035] Table 2 Two weeks after medication

[0036] The entrainer mechanism was then verified by analyzing the total flavonoid extraction rate (UV-Vis method (rutin standard), Chinese Pharmacopoeia 2020 edition), saponin retention rate (HPLC (ginsenoside Rg1), GB / T31743-2015), volatile component retention rate (GC-MS (menthone), ISO 11024-1998), and cell disruption rate (SEM observation of medicinal material microstructure, ASTM E3060-2016) of the extracts after extraction with different entrainers. The results are shown in Table 3. The only difference between Control Group 1 and Example 1 is that no entrainer was used. Control Group 2 used ethanol extraction (60% ethanol, reflux at 80°C for 2 h). Table 3 Entrainer Verification

[0037] Then group verification is carried out, and the results are shown in Table 4; Table 4 Group Verification

[0038] Example 2: The above example uses a Chinese medicine composition and a specific entrainment liquid for extraction to obtain an extract capable of treating recurrent respiratory tract infections in children. For the purpose of enzyme catalysis Boc protection and PEG-PCL micelle stability, further improvements are made on the basis of Example 1.

[0039] The hydrophobic group also strengthens the enzyme catalytic Boc protection system by coupling lipase CAL-B with Fe3O4@SiO2 to prepare Fe3O4@SiO2-CAL-B. The Fe3O4@SiO2-CAL-B in the hydrophobic group is calculated as 0.5 parts by weight. The specific preparation method is as follows: FeCl2·4H2O and FeCl3·6H2O were dissolved in deionized water at a molar ratio of 1:2 and stirred under nitrogen. NH3·H2O was then added dropwise until the pH reached 11. The mixture was reacted at 80°C for 1 hour to form Fe3O4 precipitate. The mixture was then washed with magnetic separation until neutral and vacuum dried to obtain Fe3O4 nanoparticles (10-20 nm in diameter). Fe3O4 was dispersed in an ethanol-water (4:1) solution, and TEOS (tetraethyl orthosilicate) and NH3·H2O (catalytic hydrolysis) were added. The reaction was carried out at 40°C for 6 h to form a SiO2 shell (thickness 5-10 nm). After magnetic separation and washing, Fe3O4@SiO was obtained after drying. 2； Fe3O4@SiO2 was dispersed in toluene, and APTES (3-aminopropyltriethoxysilane) was added. The reaction was carried out at 80°C for 12 hours to introduce amino groups (-NH2). Glutaraldehyde (cross-linking agent) was added to react with the amino groups of lipase CAL-B (Candida antarctica lipase B). The mixture was fixed at 25°C for 6 hours, and magnetic separation was used to remove the free enzyme to obtain Fe3O4@SiO2-CAL-B.

[0040] Instantaneous decompression is also performed during carbon dioxide extraction, specifically: The CO2 flow rate during extraction was 25 L / h, and the extraction was divided into three stages; The first stage pressure is 15 MPa, the extraction temperature is 40°C, and the extraction time is 20 min. After the first stage, the pressure is pulsed down to 5 MPa. The pressure in the second stage is 25 MPa, the extraction temperature is 35°C, and the extraction time is 30 min. During the second stage, the pressure is instantly reduced to 3 MPa and then restored every 5 min. The pressure in the third stage is 35 MPa, the extraction temperature is 32°C, and the extraction time is 40 min. After the third stage, the pressure is instantly reduced to 10 MPa. After the extraction is completed, the material is filtered and the mixed liquid is collected.

[0041] The technical solutions in the above embodiments of the present application have at least the following technical effects or advantages: Fe2+ (octahedral position) and Fe 3+ (tetrahedral / octahedral positions) by O 2- Bridging, forming a magnetically ordered arrangement, allows the magnetic domains of Fe3O4 to align in an external magnetic field, enabling rapid separation (separation efficiency > 99%). An amorphous SiO2 shell (5 nm thick) is formed via the Stöber method, providing chemical modification sites (silanol groups, -Si-OH). After hydrolysis of the silane coupling agent (APTES), -Si-O- bonds bind to the SiO2 surface, exposing terminal -NH2 groups for subsequent glutaraldehyde cross-linking. The aldehyde group (-CHO) of glutaraldehyde forms a Schiff base (C=N bond) with the amino group (-NH2) of APTES and the lysine residue (-NH2) of CAL-B. The bond energy is about 250 kJ / mol. After immobilization, the exposure rate of the active center of CAL-B (Ser-His-Asp catalytic triad) increases because the carrier restricts the closure of the α-helical "lid".

[0042] By grafting PCL (polycaprolactone), the contact angle of the support surface increased from 30° (hydrophilic) to 110° (hydrophobic). The hydrophobic environment induced the opening of the α-helical "lid" of CAL-B, reducing the binding energy of the substrate (Boc anhydride) (ΔG from -8.2 to -12.5 kJ / mol). The pores of the SiO2 shell restricted the diffusion path of the substrate, increasing the local concentration by 5 times and the reaction rate (kcat) from 120 to 180 s. -1 .

[0043] CO2 molecules (linear O=C=O) form a directional solvation layer on the SiO2 surface, reducing the interfacial tension (from 50→15 mN / m). Boc anhydride (hydrophobic, logP=2.1) is enriched in the hydrophobic area of ​​the carrier, and the local concentration is increased to 3 times that of the bulk phase. Ser of CAL-B 020 The hydroxyl group forms a hydrogen bond with the carbonyl group (C=O) of Boc anhydride (bond length 2.8Å), while the hydrophobic pocket (Ile 289 , Leu 277 ) stabilizes the substrate via van der Waals forces.

[0044] The instantaneous decompression of CO2 (35→5MPa) triggers homogeneous nucleation, generating microbubbles (diameter 1-10μm). The internal pressure of the bubbles is >100MPa. When the bubbles burst, microjets (speed >100 m / s) and shock waves are generated, destroying the cell walls of the medicinal materials (cellulose hydrogen bonds are broken). The sudden pressure change triggers CO2 turbulence, increasing the substrate velocity. The α-helix of CAL-B undergoes elastic deformation under the pressure pulse, promoting the entry of the substrate into the active center.

[0045] High pressure (35MPa) is used to improve enzyme stability and initiate the enzyme-catalyzed Boc protection reaction. Medium pressure (25MPa) is used to activate enzyme activity and low pressure (15MPa) is used to adsorb impurities. When the pressure drops suddenly, the volume of CO2 expands rapidly, generating micron-sized bubbles and turbulent shear force. The expansion process destroys material agglomeration, increases contact, and promotes the diffusion of substrates to the enzyme active site, increasing the reaction rate. The shear force flushes the inner wall of the extraction kettle, reducing the deposition of micelles or particles.

[0046] The preparation method of Example 2 was used to verify the effectiveness of the method, including enzyme activity retention rate: HPLC determination of Boc protection rate (C18 column, UV 254nm, mobile phase 0.1% TFA aqueous solution: acetonitrile = 8:2); determination of carrier recovery rate after magnetic adsorption (mass method); GC-MS (DB-5 chromatographic column, EI ion source); and determination of enzyme activity decay rate after 10 cycles. The results are shown in Table 5. Control group 3 of this example was prepared by adding only free CAL-B, and control group 4 was prepared by adding only Fe3O4@SiO2 without loading CAL-B. Table 5 Verification test of Example 2

[0047] Example 3: Example 2 strengthens the Boc protection effect by using Fe3O4@SiO2-CAL-B. In order to improve the extraction efficiency, further improvements are made on the basis of Example 2.

[0048] In the carbon dioxide extraction, pretreatment is also carried out before the first stage. The product is crushed to 100 mesh at -20°C and soaked in 5MPa liquid CO2 for 30min, with ultrasonic assistance (28kHz, 200W). Then, 5-15MPa cycle is carried out for 3 times (increase pressure by 3MPa / min and decrease pressure by 5MPa / min), which takes a total of 15min. Then, the pressure is increased step by step from 5MPa (increase pressure by 5MPa every 5min) until the pressure required for the first stage is reached.

[0049] The technical solutions in the above embodiments of the present application have at least the following technical effects or advantages: At low temperatures, water molecules form hexagonal ice crystals, which expand in volume and exert anisotropic stress on the cell wall (cellulose), causing microcracks. The growing ice crystals squeeze the cellulose chains (C-O-C bond length 1.43 Å), causing partial breakage of hydrogen bonds (bond energy ≈ 20 kJ / mol). The β-1,4 glycosidic bonds (bond energy ≈ 340 kJ / mol) are partially broken by shear forces to form oligosaccharide fragments (DP ≈ 5-10). Low temperatures cause the acyl chains of cell membrane phospholipids to change from liquid to gel, increasing their brittleness and making them more easily broken. Supercritical CO2 (density ≈0.6 g / cm³ at 5 MPa) penetrates through the pores of the medicinal materials (pore diameter > 2 nm). CO2 dissolves the cell wall lignin (non-polar component), increasing the swelling rate by 15% and the porosity simultaneously.

[0050] CO2 molecules (O=C=O) form weak hydrogen bonds (bond length ≈ 2.5Å) with hydroxyl groups (-OH) on the cellulose surface, reducing the surface energy. The sudden pressure drop triggers homogeneous nucleation, and microjets and shock waves are generated when the bubbles collapse, directly shearing the cell wall. The shock wave increases the probability of CO bonds breaking in the fibrils; hydrogen bonds and hydrophobic interactions are destroyed, and lignin (Mw ≈ 10kDa) is released from the complex.

[0051] Low-temperature micro-powders generate 100-500μm particles, with internal crack networks providing channels for CO2 penetration. CO2 pre-infiltration expands pores, and shock wave energy is transferred to molecular bonds (CO), directly promoting the release of components. After pre-infiltration, the porosity of the medicinal material increases, and the penetration depth of CO2 during the pulse stage increases. Pressure fluctuations (5↔15MPa) trigger periodic Darcy flow with a shear stress of τ≈10Pa, which peels off cell wall fragments and dissolves 5-10% of fat-soluble components (volatile oils) in the pre-infiltration stage, reducing competition for subsequent extraction; pulse fragmentation releases 20-30% of intracellular polysaccharides, forming an in-situ "extraction channel". When the pressure drops suddenly, CO2 vaporizes and absorbs heat (ΔH≈150kJ / kg), and local cooling induces thermal stress, exacerbating the brittle fracture of the cell wall.

[0052] The cell disruption rate, carbon dioxide penetration depth (physical sectioning-staining method, Nile red-0.% ethanol, carbon dioxide 5MPa staining, liquid nitrogen freezing, 10μm section, observation using confocal microscope with excitation wavelength of 543nm and emission wavelength of 590nm, penetration depth = the distance from the farthest point to the edge of the stained area), and total flavonoid extraction rate were detected. The results are shown in Table 6; Table 6 Verification test of Example 3

[0053] Example 4: Example 3 increases the CO2 penetration depth through pretreatment, but high-frequency instantaneous decompression may destroy the binding of magnetic nanoparticles (Fe3O4@SiO2) and lipase CAL-B, resulting in enzyme shedding (shedding rate 5-10%). Further improvements are made on the basis of Example 3.

[0054] After the third stage of carbon dioxide extraction, the pressure is instantly reduced to 5MPa, and a pressure-responsive enzyme loading layer is constructed on the surface of magnetic nanoparticles (Fe3O4@SiO2) to achieve directional and controllable release of the enzyme.

[0055] Specifically, the magnetic nanoparticles have an outer layer that can fall off under instantaneous reduced pressure of 5 MPa, a stable inner layer, and a middle layer Fe3O4@SiO2 that falls off at high temperature. The synthesis steps are consistent with Example 2, and then the inner layer is covalently fixed. After amination of the SiO2 surface, EDC / NHS is coupled to the lipase CAL-B (loading amount 20 mg / g), and then poly N-isopropylacrylamide (PNIPAM) is grafted, LCST = 37°C, to form a middle thermosensitive hydrogel layer, and finally, through hydrophobic modification (octadecyltrichlorosilane), the second layer of CAL-B (loading amount 10 mg / g) is adsorbed.

[0056] The technical solutions in the above embodiments of the present application have at least the following technical effects or advantages: Under instantaneous pressure reduction, the enzyme is easily peeled off and falls off. Through the pressure response of the outer layer, not only is it not fixed, but an outer layer that accelerates the shedding is also constructed to form a large amount of free enzymes, realizing the directional and controllable release of the enzymes. The shear force and bubble effect of the instantaneous pressure reduction of supercritical CO2 are used to accurately trigger the shedding of the outer layer enzyme, release the free enzyme to enhance the terminal reaction, and recover the carrier through the gradient magnetic field and reuse it. The inner layer enzyme retains its activity, realizing the maximum enzyme utilization.

[0057] Instantaneous decompression of 5MPa triggers the shedding of the outer layer enzyme (shedding rate > 90%), and the free enzyme activity is retained at 95%; after the outer layer enzyme falls off, the middle layer enzyme will gradually fall off at a temperature of 45°C, and will be shed and recovered when the inner layer needs to be recovered, protecting the inner layer enzyme from falling off during operation.

[0058] The inner layer enzyme retention rate under 35MPa high pressure is >98%, and the activity is >85% after 10 reuses.

[0059] It should be noted that, in the above embodiments, the description of each embodiment has its own focus. For parts that are not described in detail in a certain embodiment, reference can be made to the relevant descriptions of other embodiments.

[0060] Although the preferred embodiments of the present invention have been described, those skilled in the art may make additional changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0061] Obviously, those skilled in the art may make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if such changes and modifications fall within the scope of the claims and their equivalents, the present invention is intended to include such changes and modifications.

Claims

1. A Chinese medicine composition for treating recurrent respiratory tract infections in children, characterized in that: It includes 6g of Pseudostellaria baicalensis, 6g of stir-fried Atractylodes macrocephala, 10g of Poria cocos, 3g of raw licorice, 10g of raw Astragalus membranaceus, 6g of Saposhnikovia divaricata, 6g of honeysuckle, 6g of mint, 6g of Amomum villosum, 6g of Tangerine peel, 6g of Platycodon grandiflorum, 10g of Peucedanum chinense, 15g of Aster scabra, 10g of Stemona tuberosa, 6g of Citrus aurantium, and 20g of Houttuynia cordata.

2. The Chinese medicine composition for treating recurrent respiratory tract infections in children according to claim 1, characterized in that Its preparation method is: S1. Wash the formula and dry it at 60°C, crush it and sieve it with 60 mesh, then place the Chinese medicine powder in a supercritical extraction device for extraction, add entrainer to the entrainer tank, the mass ratio of powder to entrainer is 20:3, soak it for 30 minutes, and then perform supercritical carbon dioxide extraction; S2. The mixed solution was placed in a vacuum distillation apparatus, 1.2 parts of a stabilizer 6-chloronicotinic acid was added, and the mixture was stirred for 15 minutes. The pressure was set to 15 mmHg and the temperature was set to 70°C. After reacting for 2 hours, 0.4 parts of a stabilizer 6-chloronicotinic acid was added and the mixture was reacted for another 2 hours to obtain a pure extract. S3. Take 100 parts of pure extract and add it to the drum equipment. Add 3 parts of maltose, 0.5 parts of citric acid, and 5 parts of sodium chloride. The temperature is 35°C. Add 5 parts of the auxiliary agent FMOC-L-phenylalanine to the drum equipment every two hours. The drum speed is 100 r / min. React for a total of 5 hours and finally flow out to obtain the product.

3. The Chinese medicine composition for treating recurrent respiratory tract infections in children according to claim 2, characterized in that: The CO2 flow rate in supercritical carbon dioxide extraction is 25 L / h, and the extraction is divided into three stages. The first stage has a pressure of 15 MPa, an extraction temperature of 40 °C, and a time of 20 min. The second stage pressure is 25 MPa, the extraction temperature is 35°C, and the extraction time is 30 min; The pressure of the third stage is 35 MPa, the extraction temperature is 32°C, and the extraction time is 40 min. After the extraction is completed, the material is filtered and the mixed liquid is collected.

4. The Chinese medicine composition for treating recurrent respiratory tract infections in children according to claim 3, characterized in that: The entrainer comprises 0.8 parts of a hydrophobic group, 0.5 parts of a wrapping group, 0.2 parts of a separation group, and 6.0 parts of a solvent in parts by mass.

5. The Chinese medicine composition for treating recurrent respiratory tract infections in children according to claim 4, characterized in that: The hydrophobic group is prepared by mixing L-histidine and Boc anhydride in a mass ratio of 1:0.25, adding 0.1% lipase CAL-B, and reacting at 35 MPa / 35°C for 30 minutes. 3Å molecular sieves are added to adsorb the generated tert-butanol and water. The pressure is reduced to 5 MPa, and CO2 is discharged with the unreacted Boc anhydride, while the product Boc-histidine remains in the kettle.

6. The Chinese medicine composition for treating recurrent respiratory tract infections in children according to claim 4, characterized in that: The encapsulation group is prepared by mixing polyethylene glycol and polycaprolactone in a mass ratio of 4:1, adding stannous octoate as a catalyst and reacting for 6 hours at 140°C in a nitrogen environment to generate a PEG-PCL block copolymer.

7. The Chinese medicine composition for treating recurrent respiratory tract infections in children according to claim 4, characterized in that: The isolated group is Pluronic L31, which is a PEO-PPO-PEO triblock copolymer with a molecular weight of 1100 Da.

8. The Chinese medicine composition for treating recurrent respiratory tract infections in children according to claim 5, characterized in that: The hydrophobic group is further added with Fe3O4@SiO2-CAL-B, and the Fe3O4@SiO2-CAL-B in the hydrophobic group is calculated as 0.5 parts by weight. The preparation method is as follows: FeCl2·4H2O and FeCl3·6H2O were dissolved in deionized water at a molar ratio of 1:2 and stirred under nitrogen. NH3·H2O was then added dropwise until the pH reached 11. The mixture was reacted at 80°C for 1 hour to generate Fe3O4 precipitate. The mixture was then washed with magnetic separation until neutral and dried in vacuum to obtain Fe3O4 nanoparticles. Fe3O4 was dispersed in an ethanol-water (4:1) solution, and TEOS (tetraethyl orthosilicate) and NH3·H2O were added. The mixture was reacted at 40°C for 6 h to form a SiO2 shell. The mixture was then magnetically separated, washed, and dried to obtain Fe3O4@SiO2. Fe3O4@SiO2 was dispersed in toluene, APTES was added, and the mixture was reacted at 80°C for 12 h to introduce amino groups. Glutaraldehyde was added to react with the amino groups of lipase CAL-B. The mixture was fixed at 25°C for 6 h, and free enzyme was removed by magnetic separation to obtain Fe3O4@SiO2-CAL-B.

9. The Chinese medicine composition for treating recurrent respiratory tract infections in children according to claim 3, characterized in that: Instantaneous decompression is also performed during carbon dioxide extraction, specifically: In the three stages of extraction, the first stage pressure was 15 MPa, the extraction temperature was 40°C, and the extraction time was 20 min; after the first stage, the pressure was pulsed down to 5 MPa; The pressure in the second stage is 25 MPa, the extraction temperature is 35°C, and the extraction time is 30 min. During the second stage, the pressure is instantly reduced to 3 MPa and then restored every 5 min. The pressure in the third stage is 35 MPa, the extraction temperature is 32°C, and the extraction time is 40 min. After the third stage, the pressure is instantly reduced to 10 MPa. After the extraction is completed, the material is filtered and the mixed liquid is collected.

10. The Chinese medicine composition for treating recurrent respiratory tract infections in children according to claim 9, characterized in that: In the carbon dioxide extraction, pretreatment is also carried out before the first stage. The product is crushed to 100 mesh at -20℃ and soaked in 5MPa liquid CO2 for 30min, with ultrasound assistance. Then, 5-15MPa cycle is carried out 3 times, with a total time of 15min. Then, the pressure is stepped up from 5MPa to the pressure required for the first stage.

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