Preparation method of anti-allergic agent with dentinidia japonica extract wrapped by modified porous chitosan microspheres
By modifying porous chitosan microspheres to wrap Japanese dentifritic extract, the problem of traditional anti-allergic agents having a large impact on cat and dog hair and insufficient stability is solved, and efficient and long-term anti-allergic effect is achieved, which significantly improves the biological system. Utilization and stability.
Patent Information
- Application Number
- CN202411444017.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-16
- Publication Date
- 2025-01-10
AI Technical Summary
Traditional antiallergic agents have adverse effects on cat and dog hair, and are not strong insufficiency. The use of Japanese dentifrice extract alone has problems of stability and insufficient bioavailability, making it difficult to effectively allergic symptoms.
Porous chitosan microspheres were prepared by modifying chitosan, and Japanese dentifrice extract was wrapped in it to form modified porous chitosan microspheres to wrap anti-allergic agents to improve its stability and bioavailability , achieve sustained release effect.
It significantly improves the bioavailability and stability of chlorogenic acid and luteolin in Japanese dentifritic moss, achieves a long-lasting anti-allergic effect, and effectively fights allergen proteins such as house dust mites, cat dander and dog dander.
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Figure CN119280192A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to a method for preparing a pharmaceutical preparation, in particular to a method for preparing a modified porous chitosan microsphere for encapsulating an antiallergic agent of an edentula japonica extract. Background Art
[0002] As homes become more airtight, allergic symptoms such as rhinitis, conjunctivitis, bronchial asthma, and atopic dermatitis have become a social problem due to the effects of volatile organic compounds such as formaldehyde and toluene during construction, cat and dog hair produced when pets are kept indoors, and mites and mold when air conditioners are used.
[0003] Allergic reactions are immune responses caused by the body's oversensitivity to certain specific antigens. From a molecular mechanism perspective, the occurrence of allergic diseases can be divided into five steps: when a patient first comes into contact with an allergen, the patient's dendritic cells take up allergen proteins and induce the production of allergen-specific Th2 cells, which interact with B cells to induce them to produce allergen-specific IgE, which binds to mast cells to complete sensitization; when the allergen enters the mucosa again, the antigen can directly bind to the IgE on the mast cells, causing the mast cells to release a variety of bioactive mediators, which act on multiple effector organs, leading to the occurrence of allergic symptoms. The development of allergic diseases follows the atopic process, showing a trend of progressive aggravation of allergic reactions: at the beginning, an individual may only be allergic to one allergen, but as they age, the number of sensitized allergens increases, the severity of allergic symptoms increases, and allergic reactions may even transfer to another organ.
[0004] The prevention and treatment of allergic diseases can also be mainly divided into two categories: drug treatment and environmental prevention. Among them, drug treatment is aimed at the body itself; environmental prevention refers to reducing the incidence of the disease by avoiding factors that induce allergies. There are four common allergens, including dust mites, pollen, cat hair, and dog hair. Traditional antiallergic agents are not strong in antiallergic properties, and while they are antiallergic, they have adverse effects on cat and dog hair. Japanese moss is widely believed to have antiallergic effects, but there are some limitations when used alone. In order to overcome these problems, it is necessary to develop an effective drug preparation that can improve the stability, bioavailability and sustained release effect of Japanese moss extract. Summary of the invention
[0005] In view of the existing problems, the present invention first prepares modified porous chitosan microspheres by modifying chitosan, and then encapsulates the Japanese edentaria extract with the modified porous chitosan microspheres to obtain an antiallergic agent of the Japanese edentaria extract encapsulated by the modified porous chitosan microspheres.
[0006] The present invention is a method for preparing an antiallergic agent containing a modified porous chitosan microsphere encapsulating an extract of Japanese edentulous moss. The preparation method mainly comprises the following steps:
[0007] (1) adding chitosan powder to triethyl citrate, heating and stirring to prepare a solution with a certain mass ratio;
[0008] (2) adding isooctane and / or a surfactant in a certain mass ratio to the above solution, and performing emulsification treatment under certain conditions to obtain a suspension;
[0009] (3) The suspension is mixed with potassium methyl silicate in a certain mass ratio, and modified under certain conditions to obtain a modifier.
[0010] (4) freeze-drying the suspension and performing heat treatment to obtain modified porous chitosan microspheres.
[0011] (5) The porous chitosan microspheres are dissolved in double distilled water to prepare a chitosan solution of a certain concentration, a certain volume ratio of polyphosphate is added, and a certain mass ratio of Japanese moss extract is added under certain conditions to prepare modified porous chitosan microspheres encapsulating the antiallergic agent.
[0012] The heating treatment conditions described in step (1) are 120-150° C.; the stirring speed is 600-1000 rpm and the stirring time is 10-30 min; the certain mass ratio is that the mass ratio of chitosan powder to triethyl citrate is 1:(25-50). The preferred heating treatment conditions are 140-150° C.; the stirring speed is 800-1000 rpm and the stirring time is 20-30 min; the certain mass ratio is that the mass ratio of chitosan powder to triethyl citrate is 1:(25-45).
[0013] The and / or surfactant described in step (2) is Span80; and / or the mass ratio of isooctane to the surfactant is (5-15):1; the mass ratio of the above solution to isooctane and / or surfactant is (3-10):1; the certain condition is that the temperature is -5°C to 15°C and the time is 10min to 40min. Preferably, the and / or surfactant is Span80; and / or the mass ratio of isooctane to the surfactant is (10-15):1; the mass ratio of the above solution to isooctane and / or surfactant is (5-10):1; the certain condition is that the temperature is -5°C to 10°C and the time is 20min to 40min.
[0014] The certain mass ratio described in step (3) is that the ratio of the suspension to potassium methyl silicate is (10-25):1; the certain conditions are that the ultrasonic treatment power is 100-220W, the temperature is 50-80°C, the time is 30-90s, and the heating distillation time is 20-30min. Preferably, the certain mass ratio is that the ratio of the suspension to potassium methyl silicate is (15-25):1; the certain conditions are that the ultrasonic treatment power is 200-220W, the temperature is 60-80°C, the time is 30-60s, and the heating distillation time is 25-30min.
[0015] The freeze drying time in step (4) is 24-48 hours; the heat treatment condition is to heat to 350-500°C under nitrogen protection, control the heating rate at 80-100°C / h, and stay at 350-500°C for 2-4 hours. Preferably, the freeze drying time is 36-48 hours; the heat treatment condition is to heat to 450-500°C under nitrogen protection, control the heating rate at 90-100°C / h, and stay at 450-500°C for 3-4 hours.
[0016] The certain concentration described in step (5) is 4 mg / mL; the certain volume ratio is (5-15):1 of the volume ratio of chitosan solution to polyphosphate, and 1 mg / mL of polyphosphate is prepared with distilled water; the certain condition is heating at 50°C for 3-5 hours under stirring at 1000 r / min; the certain mass ratio is 1:(20-50) of the mass ratio of the Japanese moss extract to the solvent. Preferably, the certain concentration is 4 mg / mL; the certain volume ratio is (10-15):1 of the volume ratio of chitosan solution to polyphosphate, and 1 mg / mL of polyphosphate is prepared with distilled water; the certain condition is heating at 50°C for 4-5 hours under stirring at 1000 r / min; the certain mass ratio is 1:(20-40) of the mass ratio of the Japanese moss extract to the solvent.
[0017] The present invention first prepares modified porous chitosan microspheres by modifying chitosan, and then encapsulates the Japanese moss extract with the modified porous chitosan microspheres to obtain an antiallergic agent of the Japanese moss extract encapsulated by the modified porous chitosan microspheres. The antiallergic agent synthesized by the present invention can effectively improve the bioavailability and stability of chlorogenic acid and luteolin in Japanese moss, and achieve a sustained-release, long-lasting antiallergic effect. BRIEF DESCRIPTION OF THE DRAWINGS
[0018] Figure 1 This is a graph showing the anti-allergic performance of the anti-allergic agents with different proportions in Examples 1-5 of the present invention against the house dust mite allergen protein Der p 1;
[0019] Figure 2 This is a graph showing the anti-allergic performance of the anti-allergic agents with different proportions in Examples 1-5 of the present invention against the house dust mite allergen protein Der f1;
[0020] Figure 3 This is a graph showing the anti-allergic performance of the anti-allergic agents with different proportions in Examples 1-5 of the present invention against cat dander allergen protein Fel d1;
[0021] Figure 4 This is a graph showing the anti-allergic performance of the anti-allergic agents in different proportions in Examples 1-5 of the present invention against the dog dander allergen protein Can f1. DETAILED DESCRIPTION
[0022] The method of the present invention is described in detail below in conjunction with specific embodiments. This embodiment is implemented based on the technical solution of the present invention, but the protection scope of the present invention is not limited to the following embodiments.
[0023] Example 1
[0024] (1) Add 10 g of chitosan powder to 250 g of triethyl citrate, heat at 150° C., and stir at 1000 rpm for 30 min to prepare a solution with a mass ratio of 1:25;
[0025] (2) adding 52 g of a turbid solution of isooctane and Span80 in a mass ratio of 10:1 to the above solution, and emulsifying the solution at 8° C. for 30 min to obtain a suspension;
[0026] (3) The above suspension was mixed with 21 g of potassium methyl silicate, and ultrasonically treated at 220 W with the temperature set to 65° C. for 30 s, and heated for distillation for 30 min to obtain a modifier.
[0027] (4) The suspension was freeze-dried for 36 h, heated to 450° C. under nitrogen protection, the heating rate was controlled at 100° C. / h, and kept at 450° C. for 3 h for heat treatment to obtain modified porous chitosan microspheres.
[0028] (5) The porous chitosan microspheres were dissolved in double distilled water to prepare a modified porous chitosan microsphere solution with a concentration of 4 mg / mL. The polyphosphate was prepared with distilled water at a concentration of 1 mg / mL. 10 mL of the modified porous chitosan microsphere solution was mixed with 1 mL of the polyphosphate solution. Japanese moss extract with a mass ratio of 1:50 was added and mixed with the above solution. The solution was heated at 50°C for 4 h under stirring at 1000 r / min to obtain a modified porous chitosan microsphere-encapsulated antiallergic agent.
[0029] Example 2
[0030] (1) Add 10 g of chitosan powder to 250 g of triethyl citrate, heat at 150° C., and stir at 1000 rpm for 30 min to prepare a solution with a mass ratio of 1:30;
[0031] (2) adding 52 g of a turbid solution of isooctane and Span80 in a mass ratio of 9:1 to the above solution, and emulsifying the solution at 8° C. for 30 min to obtain a suspension;
[0032] (3) The above suspension was mixed with 21 g of potassium methyl silicate, and ultrasonically treated at 220 W with the temperature set to 65° C. for 30 s, and heated and distilled for 25 min to obtain a modifier.
[0033] (4) The suspension was freeze-dried for 36 hours, heated to 500°C under nitrogen protection, the heating rate was controlled at 90°C / h, and kept at 500°C for 3 hours for heat treatment to obtain modified porous chitosan microspheres.
[0034] (5) The porous chitosan microspheres were dissolved in double distilled water to prepare a modified porous chitosan microsphere solution with a concentration of 4 mg / mL. The polyphosphate was prepared with distilled water at a concentration of 1 mg / mL. 10 mL of the modified porous chitosan microsphere solution was mixed with 1 mL of the polyphosphate solution. The Japanese moss extract with a mass ratio of 1:40 was added and mixed with the above solution. The solution was heated at 50°C for 4 h under stirring at 1000 r / min to obtain a modified porous chitosan microsphere-encapsulated antiallergic agent.
[0035] Example 3
[0036] (1) Add 10 g of chitosan powder to 250 g of triethyl citrate, heat at 120° C., and stir at 1000 rpm for 30 min to prepare a solution with a mass ratio of 1:40;
[0037] (2) adding 52 g of a turbid solution of isooctane and Span80 in a mass ratio of 10:1 to the above solution, and emulsifying the solution at 8° C. for 30 min to obtain a suspension;
[0038] (3) The above suspension was mixed with 21 g of potassium methyl silicate, and ultrasonically treated at 220 W with the temperature set to 65° C. for 30 s, and heated for distillation for 30 min to obtain a modifier.
[0039] (4) The suspension was freeze-dried for 36 h, heated to 450° C. under nitrogen protection, the heating rate was controlled at 100° C. / h, and kept at 450° C. for 3 h for heat treatment to obtain modified porous chitosan microspheres.
[0040] (5) The porous chitosan microspheres were dissolved in double distilled water to prepare a modified porous chitosan microsphere solution with a concentration of 4 mg / mL. The polyphosphate was prepared with distilled water at a concentration of 1 mg / mL. 10 mL of the modified porous chitosan microsphere solution was mixed with 1 mL of the polyphosphate solution. Japanese moss extract with a mass ratio of 1:50 was added and mixed with the above solution. The solution was heated at 50°C for 4 h under stirring at 1000 r / min to obtain a modified porous chitosan microsphere-encapsulated antiallergic agent.
[0041] Example 4
[0042] (1) Add 10 g of chitosan powder to 250 g of triethyl citrate, heat at 150° C., and stir at 1000 rpm for 30 min to prepare a solution with a mass ratio of 1:45;
[0043] (2) adding 52 g of a turbid solution of isooctane and Span80 in a mass ratio of 10:1 to the above solution, and emulsifying the solution at 8° C. for 30 min to obtain a suspension;
[0044] (3) The above suspension was mixed with 21 g of potassium methyl silicate, and ultrasonically treated at 220 W with the temperature set to 65° C. for 30 s, and heated for distillation for 30 min to obtain a modifier.
[0045] (4) The suspension was freeze-dried for 36 h, heated to 450° C. under nitrogen protection, the heating rate was controlled at 100° C. / h, and kept at 450° C. for 3 h for heat treatment to obtain modified porous chitosan microspheres.
[0046] (5) The porous chitosan microspheres were dissolved in double distilled water to prepare a modified porous chitosan microsphere solution with a concentration of 4 mg / mL. The polyphosphate was prepared with distilled water at a concentration of 1 mg / mL. 10 mL of the modified porous chitosan microsphere solution was mixed with 1 mL of the polyphosphate solution. The Japanese moss extract with a mass ratio of 1:35 was added and mixed with the above solution. The solution was heated at 50°C for 4 h under stirring at 1000 r / min to obtain the modified porous chitosan microsphere-encapsulated antiallergic agent.
[0047] Example 5
[0048] (1) Add 10 g of chitosan powder to 250 g of triethyl citrate, heat at 150° C., and stir at 1000 rpm for 30 min to prepare a solution with a mass ratio of 1:50;
[0049] (2) adding 52 g of a turbid solution of isooctane and Span80 in a mass ratio of 10:1 to the above solution, and emulsifying the solution at 8° C. for 30 min to obtain a suspension;
[0050] (3) The above suspension was mixed with 21 g of potassium methyl silicate, and ultrasonically treated at 220 W with the temperature set to 65° C. for 30 s, and heated for distillation for 30 min to obtain a modifier.
[0051] (4) The suspension was freeze-dried for 36 h, heated to 450° C. under nitrogen protection, the heating rate was controlled at 100° C. / h, and kept at 450° C. for 3 h for heat treatment to obtain modified porous chitosan microspheres.
[0052] (5) The porous chitosan microspheres were dissolved in double distilled water to prepare a modified porous chitosan microsphere solution with a concentration of 4 mg / mL. The polyphosphate was prepared with distilled water at a concentration of 1 mg / mL. 10 mL of the modified porous chitosan microsphere solution was mixed with 1 mL of the polyphosphate solution. The Japanese moss extract with a mass ratio of 1:20 was added and mixed with the above solution. The solution was heated at 50°C for 4 h under stirring at 1000 r / min to obtain the modified porous chitosan microsphere-encapsulated antiallergic agent.
[0053] Test method: using Methods To detect the anti-allergic effect.
Claims
1. A method for preparing an antiallergic agent containing a modified porous chitosan microsphere encapsulating an extract of Edentula japonica, characterized in that: The preparation method mainly comprises the following steps: (1) adding chitosan powder to triethyl citrate, heating and stirring to prepare a solution with a certain mass ratio; (2) adding isooctane and / or a surfactant in a certain mass ratio to the above solution, and performing emulsification treatment under certain conditions to obtain a suspension; (3) The suspension is mixed with potassium methyl silicate in a certain mass ratio, and modified under certain conditions to obtain a modifier. (4) freeze-drying the suspension and performing heat treatment to obtain modified porous chitosan microspheres. (5) The porous chitosan microspheres are dissolved in double distilled water to prepare a chitosan solution of a certain concentration, a certain volume ratio of polyphosphate is added, and a certain mass ratio of Japanese moss extract is added under certain conditions to prepare modified porous chitosan microspheres encapsulating the antiallergic agent.
2. The method for preparing an antiallergic agent comprising modified porous chitosan microspheres encapsulating an extract of Edentula japonica according to claim 1, characterized in that: The heating treatment conditions described in step (1) are 120-150° C.; the stirring speed is 600-1000 rpm and the stirring time is 10-30 min; and the certain mass ratio is the mass ratio of chitosan powder to triethyl citrate is 1:(25-50).
3. The method for preparing an antiallergic agent comprising modified porous chitosan microspheres encapsulating an extract of Edentula japonica according to claim 1, characterized in that: The and / or surfactant described in step (2) is Span80; the certain mass ratio of and / or isooctane to the surfactant is (5-15):1; the mass ratio of the above solution to isooctane and / or surfactant is (3-10):1; the certain conditions are a temperature of -5°C to 15°C and a time of 10min to 40min.
4. The method for preparing an antiallergic agent comprising modified porous chitosan microspheres encapsulating an extract of Edentula japonica according to claim 1, characterized in that: The certain mass ratio described in step (3) is the ratio of the suspension to potassium methyl silicate (10-25):1; the certain conditions are ultrasonic treatment power of 100-220W, temperature of 50-80°C, time of 30-90s, and heating distillation time of 20-30min.
5. The method for preparing an antiallergic agent comprising modified porous chitosan microspheres encapsulating an extract of Edentula japonica according to claim 1, characterized in that: The freeze-drying time in step (4) is 24-48 hours; the heat treatment conditions are heating to 350-500°C under nitrogen protection, the heating rate is controlled at 80-100°C / h, and staying at 350-500°C for 2-4 hours.
6. The method for preparing an antiallergic agent comprising modified porous chitosan microspheres encapsulating an extract of Edentula japonica according to claim 1, characterized in that: The certain concentration described in step (5) is 4 mg / mL; the certain volume ratio is the volume ratio of chitosan solution to polyphosphate (5-15):1, and the polyphosphate is prepared with distilled water to 1 mg / mL; the certain condition is heating at 50°C for 3-5h under stirring at 1000 r / min; the certain mass ratio is the mass ratio of Japanese moss extract to solvent is 1:(20-50).