Application of food retention removing and cough relieving preparation for children in preparation of anti-helicobacter pylori medicine

Abstract

The invention belongs to the field of traditional Chinese medicines, and particularly discloses application of a food retention removing and cough relieving preparation for children in resisting helicobacter pylori. The food retention removing and cough relieving preparation for children is prepared from 10 medicinal materials including hawthorn, areca-nut, immature bitter orange, loquat leaf, fructus trichosanthis, fructus forsythiae, semen raphani, semen lepidii, cicada slough and platycodon grandiflorum, and has the effects of clearing heat, dispersing lung, removing food retention and relieving cough. Experiments prove that the food retention removing and cough relieving preparation for children has an obvious inhibiting effect on helicobacter pylori and has very effective inhibiting and killing effects on standard strains and drug-resistant strains of the helicobacter pylori in vitro. The food retention removing and cough relieving preparation for children has a wide clinical application prospect in the aspect of resisting helicobacter pylori.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine, specifically relating to the use of pediatric digestive and cough-relieving preparations in the preparation of anti-Helicobacter pylori drugs. Background Technology

[0002] Helicobacter pylori (Hp) is a spiral-shaped, microaerophilic, Gram-negative bacillus with a strong infectious capacity.

[0003] Currently, the main treatments for *Helicobacter pylori* (Hp) include standard triple therapy and bismuth quadruple therapy, both of which are eradication therapies primarily based on antibiotics. Standard triple therapy consists of one proton pump inhibitor (PPI) plus two antibiotics, with a treatment course of 7–14 days. Bismuth quadruple therapy consists of one PPI plus one bismuth agent plus two antibiotics, with a treatment course of 10 or 14 days. Commonly used antibiotics include clarithromycin, amoxicillin, metronidazole, tetracycline, furazolidone, and levofloxacin. The other treatment options are also variations of antibiotic-based therapies.

[0004] With the widespread use of these drugs, the resistance rate of *Helicobacter pylori* (Hp) to these antibiotics is slowly increasing. These drugs are generally ineffective in treating Hp infection and have significant side effects, including liver and kidney damage. H2 receptor antagonists cause varying degrees of adverse reactions such as dizziness, drowsiness, abdominal pain, and nausea. Faced with the ever-increasing proportion of drug-resistant *Helicobacter pylori* strains worldwide, and the growing difficulty in identifying new, less resistant antibiotics, there are currently few effective clinical options besides constantly changing antibiotic combinations, increasing antibiotic dosages, and extending treatment duration.

[0005] Currently, pure traditional Chinese medicine preparations for treating Helicobacter pylori infection are relatively rare, as researching more effective drugs against Helicobacter pylori is an urgent problem to be solved. Summary of the Invention

[0006] This invention provides the use of a pediatric digestive and cough-relieving preparation in the preparation of drugs or health products for the treatment or prevention of Helicobacter pylori infection.

[0007] Preferably, the pediatric cough-relieving oral liquid of the present invention comprises the following ingredients: hawthorn, areca nut, immature bitter orange, loquat leaf, trichosanthes fruit, forsythia, radish seed, lepidium seed, cicada slough, platycodon root, and other pharmaceutically acceptable excipients.

[0008] Preferably, the pediatric digestive and cough-relieving preparation is made from the following traditional Chinese medicine components:

[0009] Hawthorn 50-200 parts by weight, areca nut 50-200 parts by weight, loquat leaf 50-200 parts by weight

[0010] Citrus aurantium 50-200 parts by weight, Radish seed 80-150 parts by weight, Trichosanthes kirilowii 67-268 parts by weight

[0011] Lepidium seed 80-150 parts by weight, Platycodon root 80-150 parts by weight, Forsythia fruit 80-150 parts by weight

[0012] 50-100 parts by weight of cicada molts;

[0013] Further preferred options are:

[0014] 100 parts by weight of hawthorn, 100 parts by weight of areca nut, and 100 parts by weight of loquat leaf.

[0015] 100 parts by weight of immature bitter orange, 100 parts by weight of radish seed, and 134 parts by weight of trichosanthes fruit.

[0016] 100 parts by weight of Lepidium seed, 100 parts by weight of Platycodon root, 100 parts by weight of Forsythia fruit

[0017] Forsythia suspensa 66 parts by weight;

[0018] Furthermore, the hawthorn is stir-fried hawthorn, the loquat leaf is honey-fried loquat leaf, the radish seed is stir-fried radish seed, and the lepidium seed is stir-fried lepidium seed.

[0019] The preparation method of the pediatric digestive and cough-relieving preparation includes the following steps:

[0020] A. Add 8-10 times the amount of water to the following ingredients and decoct twice, for 1.5-2.5 hours each time: stir-fried hawthorn, areca nut, honey-fried loquat leaf, immature bitter orange, stir-fried radish seed, trichosanthes fruit, stir-fried lepidium seed, platycodon root, forsythia fruit, and cicada slough. Combine the decoctions, filter, and concentrate under reduced pressure to obtain an extract with a relative density of 1.20-1.25 at 50°C. Add 95% ethanol for precipitation to achieve an ethanol content of 60%-80%. Refrigerate for 12-48 hours, filter, and obtain the filtrate for later use.

[0021] B. Concentrate the filtrate from step A under reduced pressure to obtain an extract with a relative density of 1.30–1.35 at 50°C. Add 5–7 times the amount of water for further extraction and refrigeration for 24–48 hours. Filter the extract and concentrate it under reduced pressure to obtain an extract with a relative density of 1.25–1.30 at 50°C. Set aside for later use.

[0022] C. Take the extract from step B and prepare an oral pharmaceutical formulation directly or by adding pharmaceutically acceptable excipients through conventional processes.

[0023] Preferably, the pediatric digestive and cough-relieving preparation is one of the following: granules, oral liquid, capsules, decoction, syrup, microcapsules, and microcapsules.

[0024] Preferably, the pediatric cough and indigestion relief preparation can be used in combination with other drugs to treat or prevent Helicobacter pylori infection.

[0025] Preferably, the pediatric digestive and cough-relieving preparation of the present invention is prepared according to conventional methods in the art.

[0026] Preferably, the prescription and preparation method of the pediatric digestive and cough-relieving preparation of the present invention can be referred to the pharmacopoeia.

[0027] The beneficial effects of this invention are:

[0028] This invention provides a novel application for pediatric digestive and cough-relieving preparations in combating Helicobacter pylori. Experiments have demonstrated that it has a significant inhibitory effect on Helicobacter pylori, and the use of traditional Chinese medicine can reduce antibiotic use, thereby lowering the drug resistance rate of Helicobacter pylori. The pediatric digestive and cough-relieving oral liquid has high safety and few side effects. In addition to treating patients infected with Helicobacter pylori, it also has the therapeutic effect of regulating the spleen and stomach, addressing both the symptoms and the root cause, providing a new option for clinical medication. Attached Figure Description

[0029] Figure 1 The images show the morphological characteristics of Helicobacter pylori in the control group under different magnifications using a scanning electron microscope.

[0030] Figure 2 These are morphological images of Helicobacter pylori in the experimental group under scanning electron microscopy at different magnifications. Detailed Implementation

[0031] The present invention will be further illustrated by the following embodiments. It should be understood that the embodiments of the present invention are merely for illustrating the present invention and are not intended to limit the present invention. Therefore, any simple improvements to the present invention under the premise of the method of the present invention are within the scope of protection claimed by the present invention.

[0032] I. Source of Pediatric Digestive and Cough-Relieving Oral Liquid

[0033] Pediatric digestive and cough-relieving oral liquid, 10mL per bottle, produced by Lunan Houpu Pharmaceutical Co., Ltd., approval number: National Medicine Approval Number Z10970022.

[0034] II. In vitro antibacterial experiment of Xiaoji Zhike Oral Liquid for Children

[0035] 1. MIC value determination

[0036] Test drug: Pediatric digestive and cough syrup (10ml / vial); positive control drug: clarithromycin (MCE Company, batch number: 16542)

[0037] Strains: Helicobacter pylori standard strains ATCC43504, ATCC700392, SS1, and drug-resistant strains QYZ-001, QYZ-003, QYZ-004.

[0038] Methods: After filtering the pediatric digestive and cough-relieving oral liquid through a 0.22 μm filter membrane, multiple concentration gradients of the drug were prepared using the two-fold dilution method, including 200 μl / ml, 100 μl / ml, 50 μl / ml, 25 μl / ml, 12.5 μl / ml, and 6.25 μl / ml. Clarithromycin was used as a positive control and was dissolved in DMSO to prepare a stock solution. Before use, the solution was diluted to the corresponding concentration gradients (DMSO content ≤1%). 100 μl of the prepared drug solution was added to a 96-well plate, with at least 3 replicates for each drug concentration. Negative control wells (containing only the drug, without bacterial strain inoculation), growth control wells (containing no drug, only bacterial strain inoculation), and clarithromycin positive control wells were also included. Remove the bacterial strain from the incubator, scrape the bacterial growth onto PBS with a moistened sterile cotton swab, adjust the turbidity of the bacterial suspension to 1 McF with BHI containing 20% ​​FBS, dilute 10-fold, and inoculate into the aforementioned drug-containing 96-well plates (the final turbidity of the bacterial suspension in the 96-well plate is approximately 1 × 10⁻⁶). 6 (CFU / ml) Immediately place the 96-well plate in an incubator and incubate at 37°C under microaerophilic conditions with shaking at 150 rpm for 72 hours. Observe the results after incubation. Results are only usable when the growth control shows a button-like appearance or significant turbidity. Observe visually from bottom to top; the drug concentration at which bacterial turbidity significantly decreases is the MIC. MIC values ​​are determined within 3 generations after resuscitation, and are repeated 3 times. The inhibitory results of Xiaoji Zhike Oral Liquid on Helicobacter pylori are shown in Table 1.

[0039] Table 1.

[0040]

[0041] Note: "-" indicates no bacterial growth, and "+ to ++++" indicates bacterial growth.

[0042] Based on the above experimental results, the inhibitory effects of Xiaoji Zhike Oral Liquid on Helicobacter pylori from different sources are shown in Table 2.

[0043] Table 2.

[0044] strains ATCC43504 ATC700392 SS1 QYZ-001 QYZ-003 QYZ-004 MIC (μl / ml) 25 25 50 25 50 25

[0045] 2. Determination of MBC value

[0046] Take out the 96-well plate cultured for 72 hours in the micro-broth assay, and evenly spread 100 μL of liquid from the wells with drug concentrations of 1, 2, and 4 times the MIC onto fresh blood agar plates. After culturing for 72 hours, observe the results. The drug concentration that kills 99.9% of the bacteria is defined as the MBC. The inhibitory results of Xiaoji Zhike Oral Liquid on various Helicobacter pylori bacteria are shown in Table 3.

[0047] Table 3

[0048] strains ATCC43504 ATCC700392 SS1 QYZ-001 QYZ-003 QYZ-004 MBC (μl / ml) 25 50 50 25 50 50

[0049] As shown in Tables 1-3, Xiaoji Zhike Oral Liquid for Children exhibits good inhibitory activity against multiple sensitive / drug-resistant Helicobacter pylori strains, with MIC values ​​ranging from 25-50 μl / mL and MBC values ​​also between 25-50 μl / mL. This indicates that Xiaoji Zhike Oral Liquid for Children has a very effective inhibitory and bactericidal effect on both standard and drug-resistant Helicobacter pylori strains in vitro.

[0050] III. Electron microscopic observation of the inhibitory effect of Xiaoji Zhike Oral Liquid on Helicobacter pylori.

[0051] Using Helicobacter pylori strain ATCC700392 as an example, the microscopic morphological changes of this strain by Xiaoji Zhike oral liquid were observed using scanning electron microscopy. Bacteria cultured for 48 hours on blood agar plates were scraped off, and the turbidity of the bacterial suspension was adjusted to 1 McF using 5 mL of brain heart extract containing 10% FBS. The suspension was then placed in a tri-gas incubator and cultured at 37°C, microaerophilic environment, and 150 rpm for 24 hours with shaking. After 24 hours, 1 mL of bacterial suspension was added to 49 mL of 10% FBS brain heart extract containing the drug (the drug concentration was exactly the MIC). The growth control group was incubated with 10% FBS brain heart extract without the drug. After incubation for 10 hours in a tri-gas incubator at 37°C, microaerophilic environment, and 150 rpm with shaking, the bacterial suspension was collected into 50 mL centrifuge tubes and centrifuged at 8000 rpm, 26°C for 3 minutes. The precipitate was washed twice with 5 mL of PBS. The final precipitate was fixed with 2.5% glutaraldehyde fixative and incubated overnight at 4°C. Samples were taken from both the experimental and control groups, and images were obtained using scanning electron microscopy.

[0052] Electron microscopy images showed that the Helicobacter pylori in the control group had normal morphology, were viable, and were rod-shaped with uniform length (e.g., ...). Figure 1 In the experimental group, Helicobacter pylori showed obvious bending and inactivation, exhibiting a circular shape (e.g., ...). Figure 2 This indicates that the pediatric digestive and cough-relieving oral liquid of the present invention has a significant inhibitory effect on this strain.

[0053] In addition to oral liquids, other dosage forms of pediatric digestive and cough-relieving preparations, such as granules, capsules, decoctions, syrups, microcapsules, and microcapsules, also showed significant in vitro antibacterial effects against Helicobacter pylori.

Claims

1. The application of pediatric digestive and cough-relieving preparations in the preparation of anti-Helicobacter pylori drugs, characterized in that, The pediatric digestive and cough-relieving preparation is made from the following components: hawthorn, areca nut, immature bitter orange, loquat leaf, trichosanthes fruit, forsythia, radish seed, lepidium seed, cicada slough, platycodon root, and other pharmaceutically acceptable excipients.

2. The application according to claim 1, characterized in that, The pediatric digestive and cough-relieving preparation is made from the following traditional Chinese medicine components: Hawthorn 50-200 parts by weight, areca nut 50-200 parts by weight, loquat leaf 50-200 parts by weight Citrus aurantium 50-200 parts by weight, Radish seed 80-150 parts by weight, Trichosanthes kirilowii 67-268 parts by weight Lepidium seed 80-150 parts by weight, Platycodon root 80-150 parts by weight, Forsythia fruit 80-150 parts by weight 50-100 parts by weight of cicada molts; Further preferred options are: 100 parts by weight of hawthorn, 100 parts by weight of areca nut, and 100 parts by weight of loquat leaf. 100 parts by weight of immature bitter orange, 100 parts by weight of radish seed, and 134 parts by weight of trichosanthes fruit. 100 parts by weight of Lepidium seed, 100 parts by weight of Platycodon root, 100 parts by weight of Forsythia fruit Forsythia suspensa 66 parts by weight; Furthermore, the hawthorn is stir-fried hawthorn, the loquat leaf is honey-fried loquat leaf, the radish seed is stir-fried radish seed, and the lepidium seed is stir-fried lepidium seed.

3. The application according to claim 1, characterized in that, The preparation method of the pediatric digestive and cough-relieving preparation includes the following steps: A. Add 8-10 times the amount of water to the following ingredients and decoct twice, for 1.5-2.5 hours each time: stir-fried hawthorn, areca nut, honey-fried loquat leaf, immature bitter orange, stir-fried radish seed, trichosanthes fruit, stir-fried lepidium seed, platycodon root, forsythia fruit, and cicada slough. Combine the decoctions, filter, and concentrate under reduced pressure to obtain an extract with a relative density of 1.20-1.25 at 50°C. Add 95% ethanol for precipitation to achieve an ethanol content of 60%-80%. Refrigerate for 12-48 hours, filter, and obtain the filtrate for later use. B. Concentrate the filtrate from step A under reduced pressure to obtain an extract with a relative density of 1.30–1.35 at 50°C. Add 5–7 times the amount of water for further extraction and refrigeration for 24–48 hours. Filter the extract and concentrate it under reduced pressure to obtain an extract with a relative density of 1.25–1.30 at 50°C. Set aside for later use. C. Take the extract from step B and prepare an oral pharmaceutical formulation directly or by adding pharmaceutically acceptable excipients through conventional processes.

4. The application according to any one of claims 1-3, characterized in that, The pediatric digestive and cough-relieving preparation is one of the following: granules, oral liquid, capsules, decoction, syrup, microcapsules, and microcapsules.

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