Broad-spectrum bacteriostatic sustained-release tablet and preparation method thereof

Abstract

The present application relates to the field of broad-spectrum long-acting bacteriostatic, and relates to a broad-spectrum bacteriostatic sustained-release tablet and a preparation method thereof.The broad-spectrum bacteriostatic sustained-release tablet is composed of the following components: bacteriostatic components, attapulgite powder, polyethylene glycol and water; wherein the bacteriostatic components are a mixed solution of carvacol and cumin aldehyde or carvacol; the weight-volume ratio of the attapulgite powder to the bacteriostatic components is (10-50) g: 1 mL; and the weight ratio of the attapulgite powder to the polyethylene glycol is 2500:1.The sustained-release tablet is a green and environmentally friendly, high-efficiency broad-spectrum and long-acting natural bacteriostatic agent, which can effectively inhibit the growth of N.crassa, A.nidulans, A.fumigatus and S.eturmiunum for more than 40 days.

Description

Technical Field

[0001] This invention relates to the field of broad-spectrum long-acting antibacterial agents, and specifically to an antibacterial sustained-release tablet and its preparation method. Background Technology

[0002] Items and grains stored in damp basements and warehouses are easily contaminated by Aspergillus and other fungi, leading to damage to items or contamination of grains that render them inedible. Some cultural relics, especially those made of paper, leather, cotton, linen, and wood, are prone to microbial growth during storage. In existing technologies, the main measures taken to prevent contamination of grains or cultural relics in warehouses are: (1) temperature control: based on the characteristics of the grains or cultural relics, the temperature of the warehouse is kept below 15°C to slow down the growth rate of mold; (2) humidity control: humidity is one of the important conditions for the growth and reproduction of mold, so the relative humidity of the warehouse is kept below 60%; (3) keeping the warehouse clean and hygienic to reduce the growth of microorganisms. However, the above methods have relatively high requirements for the construction of warehouses, and microorganisms can still survive. To suppress the problem of contamination during the storage of items, it is currently urgent to develop green, environmentally friendly, efficient, and broad-spectrum natural antibacterial agents without affecting the quality of the items. Summary of the Invention

[0003] Technical issues:

[0004] This provides a green, environmentally friendly, highly efficient, broad-spectrum, and long-lasting natural antibacterial agent.

[0005] Technical concept:

[0006] Both carvacrol and cuminaldehyde are plant-derived natural products. The inventors have discovered that both carvacrol and cuminaldehyde possess broad-spectrum antibacterial activity. Attapulgite is a natural organic mineral free of harmful chemical components. Its unique three-dimensional structure allows it to adsorb and carry drug molecules, enabling slow release and sustained action, thus prolonging the duration of efficacy. Based on this, this invention uses attapulgite powder as a drug carrier to prepare broad-spectrum, long-lasting antibacterial sustained-release tablets.

[0007] Technical solution:

[0008] On the one hand, a broad-spectrum antibacterial sustained-release tablet is provided, which is composed of the following components: antibacterial component, attapulgite powder, polyethylene glycol and water; wherein, the antibacterial component is a mixture of carvacrol and cuminaldehyde or carvacrol; the weight-volume ratio of attapulgite powder to the antibacterial component is (10-50) g:1 mL; the weight ratio of attapulgite powder to polyethylene glycol is 2500:1.

[0009] On the other hand, a method for preparing an antibacterial sustained-release tablet is provided, which includes the following steps:

[0010] (1) Prepare a 30% (w / w) polyethylene glycol aqueous solution;

[0011] (2) The attapulgite powder is filtered to remove the air from the pores of the attapulgite powder.

[0012] (3) Add the antibacterial component to the attapulgite powder obtained in step (2) according to the weight-volume ratio of attapulgite powder to antibacterial component of (10-50) g: 1 mL, stir evenly, and then perform vacuum filtration so that the antibacterial component is adsorbed into the pores of the attapulgite powder to obtain mixture one; wherein, the antibacterial component is a mixture of carvacrol and cuminaldehyde or carvacrol.

[0013] (4) According to the weight ratio of attapulgite powder to polyethylene glycol of 2500:1, add the polyethylene glycol aqueous solution with a mass concentration of 30% obtained in step (1) to the mixture obtained in step (3), stir evenly to obtain mixture two, weigh a quantitative amount of mixture two and place it in a tableting machine to obtain a broad-spectrum antibacterial sustained-release tablet.

[0014] In some embodiments, step (1) specifically involves adding polyethylene glycol 20000 powder to hot water to completely dissolve it, thereby obtaining a polyethylene glycol 20000 aqueous solution with a mass concentration of 30%.

[0015] In some embodiments, in step (2) and / or step (3), the parameters of the filtration process are: to perform intermittent filtration in a cycle of “filtering for 10 seconds first, then pausing for 5 seconds” until the total cumulative filtration time is 1 minute.

[0016] In some embodiments, the antibacterial component is a mixture of carvacrol and calciferol; in the mixture of carvacrol and calciferol, the volume ratio of carvacrol to calciferol is (1-3):(1-3).

[0017] In some embodiments, the weight-to-volume ratio of attapulgite powder to antibacterial component is (10-25) g: 1 mL.

[0018] Beneficial effects:

[0019] 1. Both carvacrol and cuminaldehyde are plant-derived natural products. The inventors have discovered that both carvacrol and cuminaldehyde possess broad-spectrum antibacterial activity. Attapulgite is a natural organic mineral free of harmful chemical components. Its unique three-dimensional structure allows it to adsorb and carry drug molecules, enabling slow release and sustained action, thus prolonging the duration of efficacy. Based on this, this invention uses attapulgite powder as a drug carrier to prepare broad-spectrum, long-lasting antibacterial sustained-release tablets.

[0020] 2. This invention provides a green, environmentally friendly, highly efficient, broad-spectrum, and long-lasting natural antibacterial agent. It exhibits effective antibacterial activity against *Neurospora crassa* for over 30 days, against *A. nidulans* for over 40 days, against *A. fumigatus* for over 40 days, and against *S. eturmiunum* for over 20 days. Through further optimization of the composition of the broad-spectrum antibacterial sustained-release tablets, it is also possible to simultaneously achieve effective antibacterial activity against *Neurospora crassa*, *A. nidulans*, *A. fumigatus*, and *S. eturmiunum* for over 40 days. Attached Figure Description

[0021] Figure 1.1 The image shows the inhibitory effect of different tablets in Example 4 on Neurospora crassa, where the tablets were cultured statically at 30°C for 10 days.

[0022] Figure 1.2 The image shows the inhibitory effect of different tablets in Example 4 on Neurospora crassa, where the tablets were cultured statically at 30°C for 30 days.

[0023] Figure 2.1 The image shows the inhibitory effects of different tablets on Aspergillus nidulans in Example 5, where the tablets were cultured statically at 30°C for 10 days.

[0024] Figure 2.2 The image shows the inhibitory effects of different tablets on Aspergillus nidulans in Example 5, where the tablets were cultured statically at 30°C for 30 days.

[0025] Figure 2.3 The effect of different antibacterial components in Example 5, administered via tablets or by dripping onto filter paper, on the inhibitory effect of Aspergillus nidulans.

[0026] Figure 3.1 The image shows the inhibitory effects of different tablets in Example 6 on Aspergillus fumigatus, where the tablets were cultured statically at 30°C for 10 days.

[0027] Figure 3.2 The image shows the inhibitory effects of different tablets on Aspergillus fumigatus in Example 6, where the tablets were cultured statically at 30°C for 30 days.

[0028] Figure 3.3The effect of different antibacterial components in Example 6 on the inhibitory effect of Aspergillus fumigatus, either in tablet form or by dripping onto filter paper.

[0029] Figure 4.1 The image shows the inhibitory effects of different tablets on Stemphylium eturmiunum in Example 7, where the tablets were cultured statically at 30°C for 10 days.

[0030] Figure 4.2 The image shows the inhibitory effects of different tablets on Stemphylium eturmiunum in Example 7, where the tablets were cultured statically at 30°C for 30 days. Detailed Implementation

[0031] Example 1 - Preparation of carvacrol sustained-release tablets

[0032] A method for preparing carvacrol sustained-release tablets includes the following steps:

[0033] (1) Preparation of polyethylene glycol aqueous solution: Add polyethylene glycol 20000 (PEG20000) powder to hot water to completely dissolve the powder, so as to obtain a PEG 20000 aqueous solution with a mass concentration of 30%;

[0034] (2) The attapulgite powder is filtered (the attapulgite powder is filtered intermittently in a cycle of “filter for 10 seconds, then pause for 5 seconds” until the total cumulative filtration time is 1 minute) to remove the air from the pores of the attapulgite powder.

[0035] (3) Add carvacrol liquid to the attapulgite powder obtained in step (2) according to the weight-volume ratio of attapulgite powder to carvacrol liquid as (10-50) g: 1 mL, stir evenly, and then perform vacuum filtration (according to the procedure of “vacuum filtration for 10s, then pause for 5s”, perform intermittent vacuum filtration until the total cumulative filtration time is 1 min) to obtain a binary mixture so that the carvacrol liquid is adsorbed into the pores of the attapulgite powder.

[0036] (4) Add PEG 20000 aqueous solution with a mass concentration of 30% to the binary mixture obtained in step (3) according to the weight ratio of attapulgite powder to PEG20000 of 2500:1, stir evenly to obtain a ternary mixture, weigh out (225mg / tablet) of the ternary mixture and place it in a tableting machine to obtain carvacrol sustained-release tablets.

[0037] Example 2 - Preparation of Cuminaldehyde Sustained-Release Tablets

[0038] A method for preparing cuminaldehyde sustained-release tablets is the same as the method for preparing carvacrol sustained-release tablets in Example 1, except that carvacrol liquid is replaced with cuminaldehyde liquid.

[0039] Example 3 - Preparation of sustained-release tablets containing both carvacrol and calciferol

[0040] A method for preparing sustained-release tablets containing both carvacrol and calciferaldehyde is described, referring to the method for preparing carvacrol sustained-release tablets in Example 1, except that the liquid carvacrol is replaced with a mixture of carvacrol and calciferaldehyde, wherein the volume ratio of carvacrol to calciferaldehyde in the mixture is between 3:1 and 1:3; and the weight-volume ratio of attapulgite powder to (the mixture of carvacrol and calciferaldehyde) is (10g to 50g): 1mL.

[0041] Blank comparison example

[0042] A method for preparing tablets, referring to the method for preparing carvacrol sustained-release tablets in Example 1, differs only in that step (3) is omitted, and 30% PEG 20000 aqueous solution is added directly to the attapulgite powder obtained in step (2) according to the weight ratio of attapulgite powder to PEG20000 of 2500:1, stirred evenly to obtain a binary mixture, and quantitatively weighed (225mg / tablet) of the binary mixture and placed in a tableting machine to obtain blank tablets.

[0043] Example 4 - Inhibitory effect of different tablets on Neurospora crassa

[0044] (1) Preparation of spore suspension of Neurospora crassa

[0045] Conidia of Neurospora crassa were collected from solid culture medium using a 0.1% Tween 20 solution, counted using a hemocytometer, and diluted to a spore suspension concentration of 2 × 10⁻⁶. 6 per mL.

[0046] (2) Inhibitory effects of different antibacterial tablets on Neurospora crassa in solid culture medium

[0047] Test method: 10 μL of a 2×10⁻⁶ solution was prepared. 6 A spore suspension of 1 spore / mL was dropped into the center of a plate containing culture medium. One tablet prepared according to Examples 1 to 3 and the blank control was placed 2 cm away from the bacterial suspension. The plates were then incubated statically at 30°C for at least 40 days. Colony growth on each plate was observed. The results after 10 days are as follows: Figure 1.1 As shown, the results after 30 days are as follows: Figure 1.2As shown in Table 1, the effective inhibition time of different tablets against Neurospora crassa is summarized.

[0048] Table 1. Effective inhibition time of different tablets against Neurospora crassa (N. crassa).

[0049]

[0050] Example 5 - Inhibitory effect of different tablets on Aspergillus nidulans

[0051] (1) Preparation of Aspergillus nidulans spore suspension

[0052] Conidia of Aspergillus nidulans were collected from solid culture medium using 0.1% Tween 20 solution, counted using a hemocytometer, and diluted to a spore suspension concentration of 2 × 10⁻⁶. 6 per mL.

[0053] (2) Inhibitory effects of different antibacterial tablets on Aspergillus nidulans in solid culture medium

[0054] Test method: 10 μL of a 2×10⁻⁶ solution was prepared. 6 A spore suspension of 1 spore / mL was dropped into the center of a plate containing culture medium. One tablet prepared according to Examples 1 to 3 and the blank control was placed 2 cm away from the bacterial suspension. The plates were then incubated statically at 30°C for at least 40 days. Colony growth on each plate was observed. The results after 10 days are as follows: Figure 2.1 As shown, the results after 30 days are as follows: Figure 2.2 As shown in Table 2, the effective inhibition time of different tablets against Aspergillus nidulans is summarized.

[0055] Table 2. Effective inhibition time of different tablets against Aspergillus nidulans.

[0056]

[0057] (3) Effects of different antibacterial components on the inhibitory effect of Aspergillus nidulans by tablets or by dripping onto filter paper

[0058] Test method: 10 μL of a 2×10⁻⁶ solution was prepared. 6 A spore suspension of 1 spore / mL was dropped into the center of a plate containing culture medium. One tablet (i.e., the sustained-release tablet group) prepared in Examples 1 to 3 was placed 2 cm away from the bacterial suspension. The total volume of the antibacterial component was 60 μL, and the weight-to-volume ratio of attapulgite powder to the antibacterial component was 16.7 g:1 mL. The specific volumes of carvacrol and cuminaldehyde are shown in [the table below]. Figure 2.3 The control group used the same antibacterial component, which was directly applied to filter paper. The plates were then incubated at 30°C for 10 days, and colony growth was observed on each plate. Simultaneously, *Aspergillus nidulans* was inoculated again (10 μL of a 2×10⁻⁶ concentration). 6 A spore suspension of 1 spore per mL was dropped into the center of a plate containing culture medium and incubated statically at 30°C for 10 days. The growth of colonies on each plate was then observed.

[0059] The study found that after 10 days of culture, *Aspergillus nidulans* did not germinate in the tablet agar containing 60 μL of calciferol in the sustained-release tablet group, while *Aspergillus nidulans* began to germinate and grow in the corresponding control group (60 μL of calciferol was directly added to filter paper). After re-inoculating with *Aspergillus nidulans* and continuing static culture at 30°C for another 10 days (cumulative culture time 20 days), *Aspergillus nidulans* was found to grow in both the sustained-release tablet group and the control group, but the growth of *Aspergillus nidulans* in the control group was more vigorous and other contaminating bacteria (such as...) were also observed. Figure 2.3 (As shown).

[0060] Example 6 - Inhibitory effect of different tablets on Aspergillus fumigatus

[0061] (1) Preparation of Aspergillus fumigatus spore suspension

[0062] Conidia of *Aspergillus fumigatus* were collected from solid culture medium using 0.1% Tween 20 solution, counted using a hemocytometer, and diluted to a spore suspension concentration of 2 × 10⁻⁶. 6 per mL.

[0063] (2) Inhibitory effects of different antibacterial tablets on Aspergillus fumigatus in solid culture medium

[0064] Test method: 10 μL of a 2×10⁻⁶ solution was prepared. 6 A spore suspension of 1 spore / mL was dropped into the center of a plate containing culture medium. One tablet prepared according to Examples 1 to 3 and the blank control was placed 2 cm away from the bacterial suspension. The plates were then incubated statically at 30°C for at least 40 days. Colony growth on each plate was observed. The results after 10 days are as follows: Figure 3.1 As shown, the results after 30 days are as follows: Figure 3.2 As shown in Table 3, the effective inhibition time of different tablets against Aspergillus fumigatus is summarized.

[0065] Table 3. Effective inhibition time of different tablets against Aspergillus fumigatus.

[0066]

[0067] (3) Effects of different antibacterial components on the inhibitory effect of Aspergillus fumigatus by tablet or filter paper application

[0068] Test method: 10 μL of a 2×10⁻⁶ solution was prepared. 6 A spore suspension of 1 spore / mL was dropped into the center of a plate containing culture medium. One tablet (i.e., the sustained-release tablet group) prepared in Examples 1 to 3 was placed 2 cm away from the bacterial suspension. The total volume of the antibacterial component was 60 μL, and the weight-to-volume ratio of attapulgite powder to the antibacterial component was 16.7 g:1 mL. The specific volumes of carvacrol and cuminaldehyde are shown in [the table below]. Figure 3.3 The control group used the same antibacterial component, which was directly applied to filter paper. The plates were then incubated at 30°C for 10 days, and colony growth was observed on each plate. Simultaneously, *Aspergillus fumigatus* was inoculated again (10 μL of a 2×10⁻⁶ concentration). 6 A spore suspension of 1 spore per mL was dropped into the center of a plate containing culture medium and incubated statically at 30°C for 10 days. The growth of colonies on each plate was then observed.

[0069] Studies have shown that *Aspergillus fumigatus* did not germinate in either the sustained-release tablet group or the control group after 10 days of culture. After re-inoculation with *Aspergillus fumigatus* and continued static culture at 30°C for another 10 days (cumulative culture time 20 days), it was found that: when the dose of cuminaldehyde was 60 μL, *Aspergillus fumigatus* grew in both the sustained-release tablet group and the control group, and in the corresponding control group, not only *Aspergillus fumigatus* but also other bacteria grew; when the addition amounts of carvacrol and cuminaldehyde were 15 μL and 45 μL respectively, *Aspergillus fumigatus* did not germinate in the sustained-release tablet group, while *Aspergillus fumigatus* began to grow and reproduce in the control group (e.g., ...). Figure 3.3 (As shown).

[0070] Example 7 - Inhibitory effect of different tablets on Stemphylium eturmiunum

[0071] (1) Preparation of Spore Suspension of S. eturmiunum

[0072] Conidia of *S. eturmiunum* were collected from solid culture medium using a 0.1% Tween 20 solution. The spores were counted using a hemocytometer and diluted to a spore suspension concentration of 2 × 10⁻⁶. 6 per mL.

[0073] (2) Inhibitory effects of different antibacterial tablets on S. eturmiunum in solid culture medium

[0074] Test method: 10 μL of a 2×10⁻⁶ solution was prepared. 6A spore suspension of 1 spore / mL was dropped into the center of a plate containing culture medium. One tablet prepared according to Examples 1 to 3 and the blank control was placed 2 cm away from the bacterial suspension. The plates were then incubated statically at 30°C for at least 40 days. Colony growth on each plate was observed. The results after 10 days are as follows: Figure 4.1 As shown, the results after 30 days are as follows: Figure 4.2 As shown in Table 4, the effective antibacterial time of different tablets against S. eturmiunum is summarized.

[0075] Table 4. Effective antibacterial time of different tablets against S. eturmiunum

[0076]

Claims

1. The application of a broad-spectrum antibacterial sustained-release tablet in the preparation of a pharmaceutical agent for inhibiting fungi, characterized in that, The broad-spectrum antibacterial sustained-release tablets are composed of the following components: antibacterial ingredient, attapulgite powder, polyethylene glycol, and water; wherein the antibacterial ingredient is a mixture of carvacrol and calciferol, and the volume ratio of carvacrol to calciferol in the mixture is (1~3):(1~3); the weight-volume ratio of attapulgite powder to the antibacterial ingredient is (10~50) g:1mL; the weight ratio of attapulgite powder to polyethylene glycol is 2500:1; the fungus is Neurospora crassa (… Neurospora crassa Aspergillus nidus ( ) Aspergillus nidulans Aspergillus fumigatus () Aspergillus fumigatus ) and / or creeping mold ( Stemphylium eturmiunum ).

2. The application of the broad-spectrum antibacterial sustained-release tablet according to claim 1 in the preparation of a pharmaceutical agent for inhibiting fungi, characterized in that, The weight-to-volume ratio of the attapulgite powder to the antibacterial component is (10~25) g:1 mL.

3. The application of the broad-spectrum antibacterial sustained-release tablet according to claim 1 in the preparation of a pharmaceutical agent for inhibiting fungi, characterized in that, The preparation method of the broad-spectrum antibacterial sustained-release tablets is as follows: (1) Prepare a 30% (w / w) polyethylene glycol aqueous solution; (2) The attapulgite powder is filtered to remove the air from the pores of the attapulgite powder. (3) Add the antibacterial component to the attapulgite powder obtained in step (2) according to the weight-volume ratio of attapulgite powder to antibacterial component of (10~50) g:1 mL, stir evenly, and then perform vacuum filtration so that the antibacterial component is adsorbed into the pores of the attapulgite powder to obtain mixture one; wherein, the antibacterial component is a mixture of carvacrol and cuminaldehyde. (4) According to the weight ratio of attapulgite powder to polyethylene glycol of 2500:1, add the polyethylene glycol aqueous solution with a mass concentration of 30% obtained in step (1) to the mixture obtained in step (3), stir evenly to obtain mixture two, weigh mixture two quantitatively and place it in a tableting machine to obtain a broad-spectrum antibacterial sustained-release tablet.

4. The application of the broad-spectrum antibacterial sustained-release tablet according to claim 3 in the preparation of a pharmaceutical agent for inhibiting fungi, characterized in that, The specific step (1) is as follows: add polyethylene glycol 20000 powder to hot water to completely dissolve it, so as to obtain a polyethylene glycol 20000 aqueous solution with a mass concentration of 30%.

5. The application of the broad-spectrum antibacterial sustained-release tablet according to claim 3 in the preparation of a pharmaceutical agent for inhibiting fungi, characterized in that, In step (2) and / or step (3), the parameters for the filtration process are: to perform intermittent filtration in a cycle of "filter for 10 seconds, then pause for 5 seconds" until the total cumulative filtration time is 1 minute.

Images