Condensate powder loaded with lotus plumule total alkaloids as well as preparation method and application thereof
By using gelatin, pectin and tannin as carriers, a condensed powder was prepared to carry the total alkaloid of lotus seeds, which solved the problem that existing drugs were easily washed away, achieved efficient local treatment effects of oral mucosa, and reduced production costs.
Patent Information
- Application Number
- CN202411985163.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-31
- Publication Date
- 2025-04-04
AI Technical Summary
Existing oral mucositis treatment drugs are easily washed away by saliva or food, resulting in a reduced therapeutic effect and a lack of more suitable local oral mucosal treatment materials.
Gelatin, pectin and tannin are used as adhesion polymer carriers to carry the total alkaloid of lotus seeds to prepare a condensate powder. This powder forms nanoparticles through self-assembly reactions, which can quickly adhere to the tissue and form a hydrogel.
The aggregate powder can effectively carry the total alkali of lotus seed heart and form it through adhesion and hydrogel, which significantly improves the local therapeutic effect of the drug, reduces production costs, and is suitable for large-scale production.
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Figure CN119745809A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the field of oral mucositis therapeutic drugs, and particularly relates to a condensate powder carrying total alkaloids of lotus seed core, and a preparation method and application thereof. Background Art
[0002] Oral mucositis (OM) is a common and serious adverse reaction during cancer chemotherapy. The incidence of OM is about 20% to 40% in cancer patients receiving chemotherapy drugs. Clinically, it ranges from erythematous lesions with almost no symptoms to severe ulcers, which can cause oral pain, dysphagia, and reduced nutritional intake, thereby reducing the quality of life.
[0003] Currently, topical administration to the oral mucosa is the preferred route of administration for OM, which can both control pain / bleeding and obtain complete and rapid OM relief. In addition, topical formulations avoid the side effects associated with systemic treatments because the target tissue is easily accessible. The types of agents commonly used in clinical treatment of such oral soft tissue diseases include mouthwashes, powders, sprays, etc., which can be easily washed away by saliva or food, thereby reducing the therapeutic effect. Therefore, there is an urgent need to develop drug delivery materials that are more suitable for topical treatment of the oral mucosa. Summary of the invention
[0004] Purpose of the invention: The first purpose of the present invention is to provide a kind of agglomerate powder carrying total alkaloids from lotus seed core, the second purpose of the present invention is to provide a method for preparing the agglomerate powder carrying total alkaloids from lotus seed core, and the third purpose of the present invention is to provide the application of the agglomerate powder carrying total alkaloids from lotus seed core.
[0005] Technical solution: The lotus seed core total alkaloid-loaded agglomerate powder of the present invention comprises: gelatin, pectin, tannic acid and lotus seed core total alkaloids, wherein the gelatin, pectin and tannic acid are used as adhesive polymer carriers to load the lotus seed core total alkaloids.
[0006] Furthermore, in the adhesive polymer carrier, the mass ratio of gelatin, pectin and tannic acid is 10:1.5-2:5, preferably 10:1.5:5.
[0007] Furthermore, the loading amount of total alkaloids in the lotus seed core is 1.5-5.7%.
[0008] The preparation method of the above-mentioned lotus seed core total alkaloids-carrying aggregate powder comprises the following steps:
[0009] (1) dissolving gelatin and pectin in a solvent, stirring thoroughly to dissolve, then adding tannic acid solution, stirring again to mix, to prepare a gelatin-pectin-tannic acid mixed solution;
[0010] (2) dissolving the total alkaloids of lotus seed core in a volatile solvent to obtain a total alkaloids solution of lotus seed core;
[0011] (3) adding the lotus seed core total alkaloid solution to the gelatin-pectin-tannic acid mixed solution to carry out a self-assembly reaction to obtain a gelatin-pectin-tannic acid solution carrying the lotus seed core total alkaloid;
[0012] (4) The gelatin-pectin-tannic acid solution containing the total alkaloids of lotus seed core is pre-frozen, and then freeze-dried to obtain a coacervate powder containing the total alkaloids of lotus seed core.
[0013] Furthermore, in step (1), the solvent is water, and the stirring temperature is 60-65°C.
[0014] Furthermore, in step (1), the tannic acid solution consists of tannic acid and water, wherein the mass concentration of tannic acid is 5-10%.
[0015] Furthermore, in step (2), the volatile solvent is ethanol.
[0016] Furthermore, in step (3), the conditions of the self-assembly reaction are: reaction temperature is 25-35° C., and reaction time is 12-24 h.
[0017] Furthermore, in step (4), the freeze-drying treatment time is 24-28 hours.
[0018] The application of the above-mentioned condensate powder containing total alkaloids from lotus seed core in the preparation of medicine for treating oral mucositis.
[0019] Principle of the invention: Lotus seed is a famous traditional Chinese medicine recorded in the Compendium of Materia Medica, which is both a medicine and food and can clear away heat and detoxify. The lotus seed core is bitter and cold in nature, and its main active ingredients are alkaloids. A variety of lotus seed core alkaloids have been proven to have a broad spectrum of anti-tumor, anti-inflammatory, and antioxidant effects. The total alkaloids of lotus seed core can effectively reverse the cell viability of Hacat cells (human immortalized keratinocytes) affected by chemotherapy drugs at the cellular level.
[0020] In the present invention, gelatin, pectin and tannic acid form an adhesive polymer carrier to load the total alkaloids of lotus seed core at a certain mass ratio. The prepared agglomerate powder can achieve the adhesion of wet tissue by quickly and massively removing the hydration layer on the tissue, and can actively adapt to tissue defects of any shape.
[0021] Beneficial effects: Compared with the prior art, the present invention has the following significant effects: (1) The condensate powder (GPTA) loaded with total alkaloids from lotus seed core prepared by the present invention adopts a simple mechanical stirring method, the preparation method is simple, the materials are simple and easy to obtain, the production cost is low, and it is suitable for large-scale production; (2) The condensate powder of the present invention can not only conveniently and effectively encapsulate the total alkaloids from lotus seed core with biological activity, but also the powder can absorb the interfacial tissue fluid and buffer solution on the tissue, spontaneously form a hydrogel, and strongly adhere to the tissue through various molecular interactions, including Schiff base reaction. BRIEF DESCRIPTION OF THE DRAWINGS
[0022] Figure 1 This is the TEM image of the combination of total alkaloids and tannic acid from lotus seed core;
[0023] Figure 2 This is the particle size diagram of the total alkaloids and tannic acid in lotus seed core;
[0024] Figure 3 The ITC diagram of total alkaloids and tannic acid in lotus seed core, (a) is the original data of the titration of the two, (b) is the fitting curve and thermodynamic parameters of the titration of the two;
[0025] Figure 4 The graphs are the viscosity results of the agglomerate powders prepared under different process conditions measured on a glass slide. (a) is the viscosity measurement result of the three components, and (b) is the viscosity measurement result of the optimal ratio of the three components and the two components screened out.
[0026] Figure 5 This is the particle size diagram of the GPT solution finally screened out;
[0027] Figure 6 The graphs show the adhesive force results of GPTA powders with different drug loadings measured on glass slides and pig skins, (a) is the result on a glass slide, (b) is the result on pig skin;
[0028] Figure 7 This is the particle size diagram of the GPTA solution finally screened out;
[0029] Figure 8 is the SEM image of GPTA powder;
[0030] Fig. 9 The morphology and SEM images of the hydrogel formed by GPTA powder in water;
[0031] Fig.10 This is the result of viscosity measurement of GPTA under different conditions;
[0032] Fig.11 This is the result of measuring the viscosity of GPTA in SD rats;
[0033] Fig.12 This is an in vivo evaluation diagram of the effect of the prepared GPTA in treating oral mucositis caused by chemotherapy;
[0034] Fig.13 The ulcer healing rate of the prepared GPTA was evaluated in vivo for the treatment of chemotherapy-induced oral mucositis. DETAILED DESCRIPTION
[0035] The present invention is further described in detail below in conjunction with the examples and drawings. The test materials used in the examples can all be purchased through conventional channels.
[0036] The examples are as follows:
[0037] 1. The preparation process of the self-assembled nanoparticles of total alkaloids from lotus seed core is as follows:
[0038] The total alkaloids of lotus seed core were dissolved in ethanol to prepare a 1 mg / ml solution, which was then diluted with water to prepare a 0.1 mg / ml total alkali solution. Tannic acid was dissolved in water to prepare a 100 mg / ml solution. Tannic acid and total alkali were then mixed in a ratio of 1:6 to prepare the solution. The morphology was measured by TEM, and the particle size, potential and isothermal titration calorimetry (ITC) were measured. The results are shown in Figure 1-Figure 3 .
[0039] Results: Figure 1 As shown in the TEM image, it can be seen that the shape of the total alkali and tannic acid combined is similar to a sphere; Figure 2 As shown in the figure, the average particle size of the spheres is 370nm, the PDI is 0.16, and the potential is -13.5Mv. From the ITC results, it can be seen that the two can also react, such as Figure 3 As shown in (a) and (b) above, it can be concluded that tannic acid can carry the total alkaloids of lotus seed core and can form stable self-assembled nanobodies with it.
[0040] 2. Preparation process of gelatin-pectin-tannic acid (GPT) carrier:
[0041] Weigh 10% gelatin and 1.5% pectin, dissolve them in water, stir and dissolve at 60°C, then slowly add 5% tannic acid solution, stir for 1 hour to mix evenly, to obtain a gelatin-pectin-tannic acid solution. In the obtained solution, the mass fraction of gelatin is 10%, the mass fraction of pectin is 1.5%, and the mass fraction of tannic acid is 5%.
[0042] The final solution was pre-frozen, placed in a freeze dryer for 24 hours, and ground to obtain an adhesive polymer powder carrier (GPT).
[0043] In order to screen the appropriate process conditions, the final morphology and viscosity were used as evaluation methods. The viscosity determination method is as follows: drop a small amount of water on a glass slide, add 30 mg of the preparation powder, add a small amount of water to moisten it, cover it with another glass slide, fix it with a dovetail clip, put it in a 100°C oven for 15 minutes, leave it at room temperature for 30 minutes, and measure it on a universal testing machine (TH8203A, Suzhou Tuobo Machinery Equipment Co., Ltd.). The bonding area is 2.5 cm wide and 2.5 cm long, and the tensile test is 10 mm min -1 The bonding strength was determined by the shear strength (n=3). Figure 4 As shown in (a) and (b) in .
[0044] (1) The amounts of pectin (1.5%) and tannic acid (5%) were fixed, and the amounts of gelatin were changed to 2.5% and 5% respectively. Other operations were the same as above. As a result, the viscosity was less than when the amount of gelatin was 10%.
[0045] (2) The amounts of gelatin (10%) and tannic acid (5%) were fixed, and the amounts of pectin were changed to 3% and 6% respectively. Other operations were the same as above. As a result, the viscosity was lower than when the pectin amount was 1.5%.
[0046] (3) The amounts of gelatin (10%) and pectin (1.5%) were fixed, and the amounts of tannic acid were changed to 10% and 20% respectively. Other operations were the same as above. As a result, the viscosity was lower than when the amount of tannic acid was 5%.
[0047] (4) In order to verify the necessity of the combination of the three, two combinations were performed, the amount of gelatin 10% and pectin 1.5% was fixed, the amount of tannic acid was 0, and the other operations were the same as above. As a result, the combination of the two was too strong, and the freeze-dried state was a block that could not be ground into powder, which was not conducive to applying to the wound. The viscosity was close to that when tannic acid was present.
[0048] (5) In order to verify the necessity of the combination of the three, two combinations were performed, the amount of gelatin 10% and tannic acid 5% was fixed, the amount of pectin was 0, and the other operations were the same as above. The results showed that the combination of gelatin and tannic acid was also strong. After freeze-drying, some of them were in block form and could not be ground into powder, which was not conducive to applying to the wound.
[0049] (6) In order to verify the necessity of the combination of the three, two combinations were performed, the amount of pectin 1.5% and tannic acid 5% was fixed, the amount of gelatin was 0, and the other operations were the same as above. As a result, the combination of the two was weak and the viscosity was low.
[0050] The agglomerate powder with the best process was screened out and its particle size was measured.
[0051] Results: The average particle size of GPT is about 1 μm, and the PDI is 0.33. Figure 5 shown.
[0052] 3. Preparation process of GPTA:
[0053] Weigh 10% gelatin and 1.5% pectin and dissolve them in water, stir and dissolve them at 60°C, then slowly drop 5% tannic acid solution and stir for 1 hour to mix evenly to obtain a gelatin-pectin-tannic acid solution. Then adjust the solution temperature to 35°C, slowly add the total alkaloids of lotus seed core dissolved in ethanol to the gelatin-pectin-tannic acid solution, and evaporate the ethanol overnight to obtain a gelatin-pectin-tannic acid solution containing lotus seed core alkaloids.
[0054] The final solution was pre-frozen, and after the pre-freezing, it was placed in a freeze dryer and freeze-dried for 24 hours, so as to finally obtain the polymer powder (GPTA) loaded with lotus seed alkaloids and having adhesiveness.
[0055] Process investigation: The drug loading was set at three doses: low, medium and high, which were 1.5%, 2.9% and 5.7% respectively.
[0056] Result: The stickiness is as Figure 6 (a) and Figure 6 As shown in (b), it can be seen that the drug loading does not affect the original viscosity of GPT. A high-dose drug loading preparation was selected for subsequent experiments. The average particle size of the high-dose drug loading preparation is about 1 μm. Figure 7 As shown; the freeze-dried powder is light yellow powder, and its morphology is detected by SEM, and its morphology is as follows Figure 8 Shown are microspheres of different sizes.
[0057] 4. The condensed powder can spontaneously form a hydrogel when it comes into contact with water and has good adhesion
[0058] The coacervate powder was spread on the porcine skin tissue and a small amount of water was added. For visualization, a green dye was added.
[0059] Results: The agglomerated powder spontaneously formed a hydrogel when it encountered a small amount of water. A clear hydrogel network can also be seen from the SEM image, such as Fig. 9 It shows good adhesion and stability during stretching, bending, torsion, immersion in water and water flushing. Fig.10 shown.
[0060] 5. In vivo adhesion assay of GPTA
[0061] GPTA-Cy5: Weigh 10% gelatin and 1.5% pectin and dissolve them in water. Stir and dissolve them at 60°C. Then slowly drop 5% tannic acid solution and stir for 1 hour to mix evenly to obtain a gelatin-pectin-tannic acid solution. Then adjust the solution temperature to 35°C, slowly add the total alkaloids of lotus seed core dissolved in ethanol to the gelatin-pectin-tannic acid solution, and add 40μg / ml Cy5 to obtain a GPTA solution with Cy5.
[0062] The obtained solution was pre-frozen and then placed in a freeze dryer for 24 hours. Finally, GPTA powder with Cy5 was obtained. At the same time, a 40 μg / ml Cy5 solution was prepared. The drug adhesion was measured at different time points. Fig.11 As shown, it can be seen that the adhesion of the agglomerate powder in the rat's mouth is also very strong.
[0063] 6. Evaluation of the in vivo anti-tumor effect of GTPA in a chemotherapy-induced oral mucositis model
[0064] (1) Experimental animals: SD rats, male, 6 weeks old, weighing 180-200 g
[0065] (2) Dosage regimen and grouping: SD rats were adaptively raised for 3 days after purchase, and then intraperitoneally injected with 5-FU 30 mg / kg on days 1, 3, 5, and 7. On day 9, they were anesthetized with isoflurane, and the labial mucosa of the rat's lower incisor was dried with a sterile cotton ball. Then, a filter paper with a diameter of 4 mm was dipped in 50% acetic acid and treated on the rat's gingival mucosa for 60 seconds. The excess acetic acid was wiped off with a cotton ball, and an ulcer formed after 24 hours. The rats were randomly divided into 5 groups: 1) control group: PBS; 2) model group; 3) positive drug group: Episil; 4) marketed drug Lianxin capsule group; 5) GPTA group. Treatment began on day 10. The GPTA group and Lianxin capsule group were given 20 mg each time, and the positive drug was given once a spray each time, three times a day, for 12 consecutive days. During this period, 5-FU 30 mg / kg was intraperitoneally injected every other day to maintain the concentration of chemotherapy drugs. On days 1, 4, 7, 10, and 12, photos were taken to record the ulcer conditions, and the size of the oral ulcers in the rats was recorded. Determination of ulcer area size: The length (L) and width (W) of the ulcer were measured with a vernier caliper, and the rat ulcer volume was calculated according to the following formula: ulcer area = L × W / 2, ulcer healing rate = (ulcer area on day 0 - ulcer area on day n) / ulcer area on day 0.
[0066] Results: Fig.12 and Fig.13 As shown, both the positive drug group and the GPTA group had a good therapeutic effect on oral mucositis induced by chemotherapy.
Claims
1. A powder containing total alkaloids from lotus seed core, characterized in that: The agglomerate powder comprises gelatin, pectin, tannic acid and total alkaloids of lotus seed core, wherein the gelatin, pectin and tannic acid are used as adhesive polymer carriers to load the total alkaloids of lotus seed core.
2. The agglomerate powder according to claim 1, characterized in that In the adhesive polymer carrier, the mass ratio of gelatin, pectin and tannic acid is 10:1.5-2:
5.
3. The agglomerate powder according to claim 1, characterized in that: The loading amount of the total alkaloids of lotus seed core is 1.5-5.7%.
4. A method for preparing the aggregate powder containing total alkaloids from lotus seed core according to claim 1, characterized in that: The following steps are involved: (1) dissolving gelatin and pectin in a solvent, stirring thoroughly to dissolve, then adding tannic acid solution, stirring again to mix, to prepare a gelatin-pectin-tannic acid mixed solution; (2) dissolving the total alkaloids of lotus seed core in a volatile solvent to obtain a total alkaloids solution of lotus seed core; (3) adding the lotus seed core total alkaloid solution to the gelatin-pectin-tannic acid mixed solution to carry out a self-assembly reaction to obtain a gelatin-pectin-tannic acid solution carrying the lotus seed core total alkaloid; (4) The gelatin-pectin-tannic acid solution containing the total alkaloids of lotus seed core is pre-frozen, and then freeze-dried to obtain a coacervate powder containing the total alkaloids of lotus seed core.
5. The preparation method according to claim 4, characterized in that: In step (1), the solvent is water and the stirring temperature is 60-65°C.
6. The preparation method according to claim 4, characterized in that: In step (1), the tannic acid solution consists of tannic acid and water, wherein the mass concentration of tannic acid is 5-10%.
7. The preparation method according to claim 4, characterized in that: In step (2), the volatile solvent is ethanol.
8. The preparation method according to claim 4, characterized in that: In step (3), the conditions of the self-assembly reaction are: reaction temperature is 25-35° C., and reaction time is 12-24 h.
9. The preparation method according to claim 4, characterized in that: In step (4), the freeze-drying treatment time is 24-28 hours.
10. Use of the agglomerate powder containing total alkaloids from lotus seed core according to any one of claims 1 to 3 in the preparation of a medicament for treating oral mucositis.