Application of Thonningianin A in bacteriostasis and bacteriostatic agent

Abstract

The invention discloses application of Thonningianin A in a bacteriostatic agent and the bacteriostatic agent, fills up the blank of Thonningianin A research in the bacteriostatic direction, and has important theoretical value and clinical significance.

Description

Technical Field

[0001] The invention relates to a new use of a compound, and in particular to an application of Thonningianin A in an antibacterial agent and the antibacterial agent. Background Art

[0002] Pickembrin (PCP) is a traditional Chinese herbal medicine. Various active extracts of PCP have been identified. Among them, Thonningianin A (TA), which stands for pickembrin dihydrochalcone-7-O-[3″-O-galloyl-4″,6″-(S)-hexahydroxydiphenyl]-glucoside, is a new monomer compound extracted from PCP. There have been many studies on TA, but no research has involved antibacterial effects.

[0003] The most important function of the skin is to prevent biological dehydration and protect the internal structure from environmental influences. When the wound is infected, bacteria will cause a persistent inflammatory response at the site of infection, further delaying the healing process, and in severe cases may lead to complications such as sepsis. The high-sugar microenvironment of diabetic wounds is more susceptible to bacterial infection, resulting in slow healing of diabetic wounds. Therefore, finding better antibacterial strategies has become a topic of great concern. Summary of the invention

[0004] In view of this, the present invention provides an application of Thonningianin A in antibacterial treatment and an antibacterial agent.

[0005] The technical solution is as follows:

[0006] The first aspect of the present invention provides a use of Thonningianin A in antibacterial treatment.

[0007] A second aspect of the present invention provides a use of Thonningianin A in the preparation of an antibacterial agent.

[0008] Preferably, the bacteriostatic agent is used to inhibit the activity of Gram bacteria.

[0009] Preferably, the Gram's bacteria include Escherichia coli and / or Staphylococcus aureus.

[0010] The third aspect of the present invention provides an antibacterial agent, wherein the active ingredient of the antibacterial agent includes Thonningianin A.

[0011] Preferably, the active ingredient of the antibacterial agent is Thonningianin A.

[0012] Preferably, the antibacterial agent is an oral agent or an external application agent.

[0013] Preferably, the antibacterial agent is used to promote delayed wound healing caused by or associated with diabetes.

[0014] Preferably, the external application agent further comprises a carrier and an acceptable excipient, wherein the carrier is chitosan, and the excipient is an aqueous solution of a polyoxyethylene-polyoxypropylene ether triblock copolymer.

[0015] Preferably, the dressing is a 15-25% (w / v) aqueous solution of polyoxyethylene polyoxypropylene ether triblock copolymer. When the temperature is <20°C, the external application agent is in an aqueous solution state. When the temperature is ≥20°C, the external application agent changes into a gel state.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] The present invention explores the antibacterial effect of Thonningianin A (TA) through antibacterial experiments and finds that TA can inhibit the growth of Gram-positive bacteria-Staphylococcus aureus (S. aureus 25923 TM )) and Gram-negative bacteria - Escherichia coli (E.coli 25922 TM ) Therefore, it has a good effect on the difficult healing of wounds caused by infections caused by these two types of bacteria, especially it can promote the healing of diabetic wounds combined with bacterial infections. The present invention provides a new direction for the treatment of chronic diabetic wounds and has important theoretical value and clinical significance. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 is the phase diagram of the inhibition zone;

[0019] Figure 2 This is a statistical chart of the colony area of ​​Escherichia coli and Staphylococcus aureus;

[0020] Figure 3 This is a diagram showing the changes in the wounds of rats. DETAILED DESCRIPTION

[0021] The present invention is further described below in conjunction with embodiments and drawings.

[0022] The term "and / or" in the specification and claims is used to describe the association relationship of associated objects, indicating that three relationships may exist. For example, A and / or B can represent: A exists alone, A and B exist at the same time, and B exists alone. The character " / " generally indicates that the associated objects are in an "or" relationship.

[0023] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present application. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, without contradiction.

[0024] In the following examples and comparative examples, unless otherwise specified, the reagents or instruments used without indicating the manufacturer are all conventional products that can be purchased from the market. If no specific conditions are indicated, conventional conditions or conditions recommended by the manufacturer are followed.

[0025] 1. Experimental materials and instruments:

[0026] 1. Chitosan (CS, deacetylation degree ≥95%, viscosity 100-200 MPa / s), purchased from Aladdin (Shanghai, China), item number: C105799-2.5 kg;

[0027] 2. Acetic acid (1 wt%), purchased from Beijing Qisong Biotechnology Co., Ltd., item number: BQS116462-500ml;

[0028] 3. Sodium hydroxide solution (NaOH, c = 2 mol / L), purchased from Shenzhen Bolinda Technology Co., Ltd., item number: SLT-E11D1GAS;

[0029] 4. Sodium tripolyphosphate (TPP), purchased from Sigma-Aldrich, catalog number: 238503;

[0030] 5. Pluronic(R) F-127, purchased from Sigma-Aldrich, catalog number: P2443-250G;

[0031] 6. Phosphate buffered saline (PBS, pH 7.4), purchased from Gibco, catalog number: C10010500BT;

[0032] 7. Thonningianin A (TA), purchased from Chengdu Pusi Biotechnology Co., Ltd., catalog number: PS0556-0020;

[0033] 8. 60% high-fat feed: purchased from Jiangsu Collaborative Pharmaceutical Bioengineering Co., Ltd., product number: D12492;

[0034] 9. SPF grade rat maintenance feed was purchased from Huanyu Biotechnology Co., Ltd.

[0035] 10. SD rats, each weighing about 200 g, were purchased from the Animal Experiment Center of Southwest Medical University;

[0036] 11. Streptozotocin (STZ), purchased from Sichuan Philbos Technology Co., Ltd., product number: S8050-100mg;

[0037] 12. Aike test strips, including instruments: purchased from Benyuansheng Experimental Equipment Business Department, Jiangyang District, Luzhou City, item number: EZ3-200;

[0038] 13. Polyoxyethylene polyoxypropylene ether triblock copolymer (PF-127) was purchased from Sigma-aldrich Company, product number: P2443.

[0039] 2. Preparation of external application medicine

[0040] Preparation Example 1

[0041] 1. Preparation of TA-CNPS nanoparticles: 45 mg of chitosan was dissolved in 15 ml of acetic acid solution (1 wt%). In order to make the chitosan fully and evenly mixed and dissolved with the acetic acid solution, it was slowly added at a speed of 22.5 mg / min, and stirred at a speed of 600 rpm until dissolved. Then, the pH value of the obtained solution was adjusted to 5.5±0.5 with 2 mol / L NaOH to obtain a chitosan solution;

[0042] A mixed solution of PEG400, physiological saline and anhydrous ethanol is prepared, wherein the volume ratio of PEG400, physiological saline and anhydrous ethanol is 6:3:1 to obtain a second solution;

[0043] Prepare a 0.1% (w / v) sodium tripolyphosphate solution (the fourth solution) with distilled water;

[0044] 9 mg of TA was dissolved in 15 ml of the second solution to form a TA solution, and then the obtained TA solution was mixed with the chitosan solution to obtain a third solution, and the fourth solution was added dropwise to the third solution at a dropping speed of 40 drops / min. A milky white suspension began to appear in the mixed solution, 15 ml of sodium tripolyphosphate solution was added dropwise, and then the obtained liquid was centrifuged at 1000 rpm for 30 minutes. The obtained solid particles were washed three times with deionized water and then freeze-dried at -80°C for 72 hours to obtain chitosan nanoparticles embedded with TA, recorded as TA-CNPS nanoparticles, which were stored in a dark environment at -20°C for later use.

[0045] 2. Preparation of TA-CNPS-PF hydrogel: Dissolve PF-127 in 4°C distilled water, stir magnetically to form 45000 μL of a 20% (w / v) PF-127 clear solution, and then disperse all the TA-CNPS nanoparticles obtained in step 1 in the PF-127 clear solution at 4°C to obtain a TA-CNPS-PF hydrogel, which is stored at 4°C for later use. In the TA-CNPS-PF hydrogel prepared in this preparation example, the mass ratio of chitosan to TA is 1:0.2, and the volume mass ratio of the polyoxyethylene polyoxypropylene ether triblock copolymer aqueous solution to TA is 100 μL:0.02 mg.

[0046] The obtained TA-CNPS-PF hydrogel is in a flowing aqueous solution state when the temperature is less than 20°C, and becomes a non-flowing gel state when the temperature is ≥20°C.

[0047] Preparation Example 2

[0048] This preparation example adopts the same method as that of Preparation Example 1, except that 18 mg of TA is dissolved in 15 ml of the second solution to form a TA solution. In the TA-CNPS-PF hydrogel prepared in this preparation example, the mass ratio of chitosan to TA is 1:0.4, and the volume mass ratio of the polyoxyethylene polyoxypropylene ether triblock copolymer aqueous solution to TA is 100 μL:0.04 mg.

[0049] Preparation Example 3

[0050] This preparation example adopts the same method as that of Preparation Example 1, except that 27 mg of TA is dissolved in 15 ml of the second solution to form a TA solution. In the TA-CNPS-PF hydrogel prepared in this preparation example, the mass ratio of chitosan to TA is 1:0.6, and the volume mass ratio of the polyoxyethylene polyoxypropylene ether triblock copolymer aqueous solution to TA is 100 μL:0.06 mg.

[0051] Preparation Example 4

[0052] This preparation example adopts the same method as that of Preparation Example 1, except that 36 mg of TA is dissolved in 15 ml of the second solution to form a TA solution. In the TA-CNPS-PF hydrogel prepared in this preparation example, the mass ratio of chitosan to TA is 1:0.8, and the volume mass ratio of the polyoxyethylene polyoxypropylene ether triblock copolymer aqueous solution to TA is 100 μL:0.08 mg.

[0053] Preparation Example 5

[0054] This preparation example adopts the same method as that of Preparation Example 1, except that TA is not dissolved in the second solution.

[0055] Preparation Example 6

[0056] PF-127 was dissolved in 4°C distilled water and stirred magnetically to form a 20% (w / v) PF-127 clear solution.

[0057] 3. Drug concentration screening

[0058] 1. Diabetic rat modeling:

[0059] Preparation of 1% streptozotocin solution: Dissolve streptozotocin (STZ) in 0.1 mol / L sodium citrate buffer, freshly prepare a 10 mg / mL STZ solution, and filter and sterilize it with a 0.22 μm filter. Prepare it before use and protect it from light.

[0060] Six SD rats were divided into two groups, one of which was a normal group and the other five were diabetic groups.

[0061] Normal group: After being fed with SPF grade mice maintenance diet for 4 weeks, the rats were fasted (but not water-deprived) for 12 hours, and each rat was given 0.9% saline 30 mg / kg intraperitoneal injection for 4 consecutive days, and then continued to be fed with SPF grade diet for 2 weeks;

[0062] Diabetes group: 5 rats were fed 60% high-fat diet for 4 weeks, fasted (but not water) for 12h, and each rat was given 1% streptozotocin solution 30mg / kg intraperitoneal injection for 4 consecutive days, and then continued to be fed with 60% high-fat diet for 2 weeks.

[0063] Then, a circular skin incision with a diameter of 2 cm was made at the junction of the neck and back of all rats, deep to the fascia layer.

[0064] 2. Drug concentration screening: The wounds of rats in the normal group (Ctrl) were not coated with drugs; among the 5 rats in the diabetic group, one of them was used as the diabetic control group (DM), and its wounds were not treated in any way. The wounds of the other four rats were evenly coated with 200 μL of TA-CNPS-PF hydrogel prepared in Preparation Examples 1-4. After coating, the TA-CNPS-PF hydrogel in the aqueous solution state quickly changed into a non-flowable gel state and adhered firmly to the wound surface. Based on the average weight of 200 g of rats, the dosage of rats coated with Examples 1-4 reached 0.2 mg / Kg, 0.4 mg / Kg, 0.6 mg / Kg, and 0.8 mg / Kg, respectively.

[0065] The dressing was changed every two days, and photos were taken at 0D, 3D, 7D, and 14D.

[0066] 3. Analysis of drug concentration screening results: The random blood glucose, body weight and wound healing of rats were monitored respectively. The blood glucose level of rats was measured by tail vein blood sampling. The results are as follows.

[0067] Table 1 Blood glucose monitoring values

[0068]

[0069] As can be seen from Table 1 , the random blood glucose values ​​of rats on 3 days after STZ injection were all ≥16.7 mmol / L.

[0070] Table 2 Weight monitoring

[0071]

[0072] As can be seen from Table 2, the body weight of the normal diet group increased after injection of saline and continued feeding for 14 days, while the body weight of the control group and the diabetic model group decreased after injection of STZ and continued feeding for 14 days.

[0073] The criteria for successful modeling of type 2 diabetes are: after injection of STZ, the 3D random blood glucose is ≥16.7mmol / L and the rats show polydipsia, polyphagia, polyuria, dull yellow fur, decreased activity, and weight loss, which is considered a successful modeling of type 2 diabetes. It can be seen that the rats fed with the modeling method have successfully modeled diabetes.

[0074] The changes of rat wounds Figure 3 shown.

[0075] from Figure 3In the figure, the four photos from top to bottom in the first column are the wound photos of the normal diet group (denoted as Ctrl) rats on the 0th, 3rd, 7th and 14th days, the four photos from top to bottom in the second column are the wound photos of the diabetic control group (denoted as DM) rats on the 0th, 3rd, 7th and 14th days, the four photos from top to bottom in the third column are the wound photos of the diabetic model group (denoted as 0.2 mg / Kg TA-CNPS-PF) rats with a TA dosage of 0.2 mg / Kg (denoted as 0.2 mg / Kg TA-CNPS-PF) on the 0th, 3rd, 7th and 14th days, the four photos from top to bottom in the fourth column are the wound photos of the diabetic model group (denoted as 0.4 mg / Kg TA-CNPS-PF) rats with a TA dosage of 0.4 mg / Kg The four photos from top to bottom in the fifth column are wound photos of rats in the diabetic model group (recorded as 0.6 mg / Kg TA-CNPS-PF) with a dosage of 0.6 mg / Kg based on TA at 0D, 3D, 7D and 14D, and the four photos from top to bottom in the sixth column are wound photos of rats in the diabetic model group (recorded as 0.8 mg / Kg TA-CNPS-PF) with a dosage of 0.8 mg / Kg based on TA at 0D, 3D, 7D and 14D.

[0076] from Figure 3 It can be seen that the wound healing on the 3rd and 7th days was not obvious. On the 14th day, compared with the normal diet group, the wound healing rate of the diabetic control group was the lowest, and the wound healing rate of the model group with a TA dosage of 0.6 mg / Kg was the highest. The wound healing rates of the three model groups with TA dosages of 0.2 mg / Kg, 0.4 mg / Kg and 0.8 mg / Kg were higher than those of the normal diet group and the diabetic control group. Therefore, the TA dosage of 0.6 mg / Kg was the most further experimental concentration.

[0077] Since this study is a continuation of the prior application, this section regarding modeling, blood glucose testing, and drug concentration screening is the same as that of the prior application CN118453628A.

[0078] IV. In vitro antibacterial experiment

[0079] 1. Preparation of Tryptic Soy Agar (TSA):

[0080] Weigh 30.0 g of tryptone soy broth, heat and stir to dissolve in 1000 ml of distilled water, then weigh 10 g of agar powder, stir and heat again until dissolved to obtain culture medium, dispense the culture medium into bacterial culture dishes, and after solidification, sterilize at 121°C and high temperature and high pressure for 20-30 minutes before use.

[0081] 2. Inoculation of bacteria:

[0082] E. coli 25922 TM ) and Staphylococcus aureus 25923 TM ) in sterile liquid culture medium and calibrated to a 0.5 McFarland turbidimetric standard (1x10 8 CFU / mL), and then use a bacterial spreader inoculator (model: XK) to evenly distribute the bacteria on the surface of the tryptic soy agar (TSA) prepared in step (1).

[0083] The groups were divided into groups according to the following requirements. After the corresponding external application agent samples of each group were sterilized by ultraviolet light for 15 minutes, they were dropped into the center of the above bacterial culture dish with a diameter of 1.5 cm. After culturing at 37°C under aerobic conditions for 24 hours, photos were taken. The specific groups were as follows:

[0084] ① Control (NC) group, without any external application agent;

[0085] ② Normal saline (NS) group, 50 μL of 0.9% normal saline was added;

[0086] ③ Hydrogel (PF) group, add 50 μL of the topical preparation prepared in Preparation Example 6

[0087] ④CNPS-PF group, 50 μL of the external application agent prepared in Preparation Example 5 was added dropwise.

[0088] ⑤TA-CNPS-PF group, 50 μL of the external application agent prepared in Preparation Example 3 was added dropwise.

[0089] The experimental results are as follows Figure 1 As shown, the first line is the Gram-negative bacteria-Escherichia coli antibacterial experiment, and the five photos from left to right are the control group (NC), saline group (NS), hydrogel group (PF), nanoparticle-hydrogel group (CNPS-PF), and TA-nanoparticle-hydrogel group (0.6mg / Kg TA-CNPS-PF). The second line is the Gram-positive bacteria-Staphylococcus aureus antibacterial experiment, and the five photos from left to right are the control group (NC), saline group (NS), hydrogel group (PF), nanoparticle-hydrogel group (CNPS-PF), and TA-nanoparticle-hydrogel group (0.6mg / Kg TA-CNPS-PF). It can be seen that the TA-CNPS-PF group shows obvious in vitro antibacterial properties, and its antibacterial ability significantly exceeds that of PF.

[0090] like Figure 2 As shown, Figure 2(a) is a statistical graph of the colony area of ​​Escherichia coli. The area occupied by the colony of Escherichia coli in the bacterial culture dish was calculated by ImageJ software. It can be seen from the statistical graph that the colony area of ​​Escherichia coli in the control group (NC) and the normal saline group (NS) is 45%, the colony area of ​​Escherichia coli in the hydrogel group (PF) is 40%, the colony area of ​​Escherichia coli in the nanoparticle-hydrogel group (CNPS-PF) is 41%, and the colony area of ​​TA-nanoparticle-hydrogel group (0.6 mg / Kg TA-CNPS-PF) is 24%. It can be concluded that hydrogel can inhibit the growth of Escherichia coli, and the TA-nanoparticle-hydrogel group (0.6 mg / Kg TA-CNPS-PF) can significantly inhibit the growth of Escherichia coli. By comparing the nanoparticle-hydrogel group (CNPS-PF) with the TA-nanoparticle-hydrogel group (0.6 mg / Kg TA-CNPS-PF), it can be analyzed that TA has the effect of inhibiting the growth of Escherichia coli.

[0091] like Figure 2 (b) shows a statistical graph of the colony area of ​​Staphylococcus aureus. The area occupied by the colony of Staphylococcus aureus in the bacterial culture dish was calculated by ImageJ software. It can be seen from the statistical graph that the colony area of ​​Staphylococcus aureus in the control group (NC) and the normal saline group (NS) is 95%, the colony area of ​​Staphylococcus aureus in the hydrogel group (PF) is 60%, the colony area of ​​Staphylococcus aureus in the nanoparticle-hydrogel group (CNPS-PF) is 57%, and the colony area of ​​Staphylococcus aureus in the TA-nanoparticle-hydrogel group (0.6 mg / Kg TA-CNPS-PF) is 51%. It can be concluded that hydrogel can inhibit the growth of Staphylococcus aureus, and the TA-nanoparticle-hydrogel group (0.6 mg / Kg TA-CNPS-PF) can significantly inhibit the growth of Staphylococcus aureus. By comparing the nanoparticle-hydrogel group (CNPS-PF) with the TA-nanoparticle-hydrogel group (0.6 mg / Kg TA-CNPS-PF), it can be analyzed that TA has the effect of inhibiting the growth of Staphylococcus aureus.

[0092] In summary, TA has the effect of inhibiting Escherichia coli and Staphylococcus aureus. (* represents P<0.05, **P<0.01, ***P<0.001, ****P<0.0001), which is statistically significant. In particular, the combination of TA and hydrogel can effectively enhance the antibacterial effect.

[0093] IV. In vivo experiments verified that TA-CNPS-PF can promote the healing of diabetic infection wounds

[0094] 1. Construction of diabetic wound model

[0095] According to the experimental needs, a diabetic rat wound model was established, which was divided into a normal group and a diabetic model group. The feeding method and modeling method of the normal group and the model group were the same as those in the drug concentration screening experiment.

[0096] 2. Construction of diabetic wound bacterial infection model

[0097] Normal group (Ctrl): rats fed normally, without any bacterial colonies inoculated after wound construction;

[0098] Normal + E. coli group: rats fed normally, inoculated with E. coli after wound construction;

[0099] Normal + Staphylococcus aureus group: rats fed with normal diet were inoculated with Staphylococcus aureus after wound construction;

[0100] Diabetes group: rats with successful establishment of diabetes, without any bacterial colonies inoculated after wound establishment;

[0101] Diabetes + E. coli group: rats with successful diabetes were inoculated with E. coli after wounds were established;

[0102] Diabetes + Staphylococcus aureus group: rats with successful diabetes were inoculated with Staphylococcus aureus after wounds were established;

[0103] Diabetes+E. coli+CNPS-PF group: rats with successful diabetes were inoculated with E. coli after wounds were established;

[0104] Diabetes+Staphylococcus aureus+CNPS-PF group: rats with successful diabetes were inoculated with Staphylococcus aureus after wounds were established;

[0105] Diabetes+E. coli+TA-CNPS-PF group: rats with successful diabetes were inoculated with E. coli after wounds were established;

[0106] Diabetes+Staphylococcus aureus+TA-CNPS-PF group: rats with successful diabetes were inoculated with Staphylococcus aureus after wounds were established;

[0107] Among them, all infection groups that needed to be inoculated with Escherichia coli or Staphylococcus aureus were dripped with 10 6 Colony forming units (CFU) / ml corresponding to bacterial colonies.

[0108] 3. Drug administration

[0109] No drug was administered to the wounds of rats in the normal group, normal + E. coli group, normal + Staphylococcus aureus group, diabetes group, diabetes + E. coli group, and diabetes + Staphylococcus aureus group;

[0110] Each other group of rats was divided into groups and dosed as follows:

[0111] The diabetes + E. coli + CNPS-PF group and the diabetes + Staphylococcus aureus + CNPS-PF group were used as drug blank control groups and coated with the gel prepared in Preparation Example 5; the diabetes + E. coli + TA-CNPS-PF group and the diabetes + Staphylococcus aureus + TA-CNPS-PF group were used as treatment groups and coated with the TA-CNPS-PF hydrogel prepared in Preparation Example 3 at a ratio of 0.6 mg / Kg. The gel coating amount of all groups was 200 μL / time / mouse.

[0112] 4. Results Analysis

[0113] The study found that the antibacterial rate of the treatment group treated with TA-CNPS-PF hydrogel was not less than 60%, preferably not less than 70%, and may even be 80%, 90%, or even not less than 95%, and can reach up to 99% before and after treatment, regardless of whether it was Escherichia coli or Staphylococcus aureus infection. The wound healing speed of the rats in this group was fast, and the healing rate was close to or higher than that of the normal group, and was significantly higher than that of the diabetes + Escherichia coli (or Staphylococcus aureus) + CNPS-PF group and the diabetes group, while the wound healing rate of the diabetes group was the lowest.

[0114] This shows that TA has the effect of inhibiting Escherichia coli and Staphylococcus aureus, and thus promotes the healing of wounds by inhibiting bacteria. In addition, this conclusion was also confirmed by observing the morphological changes of bacteria through electron microscopy.

[0115] Finally, it should be noted that the above description is only a preferred embodiment of the present invention. Under the guidance of the present invention, ordinary technicians in this field can make various similar expressions without violating the purpose and claims of the present invention, and such changes all fall within the scope of protection of the present invention.

Claims

1. Application of Thonningianin A in antibacterial activities.

2. Application of Thonningianin A in the preparation of antibacterial agents.

3. The use according to claim 1, characterized in that: The bacteriostatic agent is used to inhibit the activity of Gram bacteria.

4. The use according to claim 3, characterized in that: The Gram bacteria include Escherichia coli and / or Staphylococcus aureus.

5. An antibacterial agent, characterized in that The active ingredient of the antibacterial agent includes Thonningianin A.

6. The antibacterial agent according to claim 5, characterized in that The active ingredient of the antibacterial agent is Thonningianin A.

7. The antibacterial agent according to claim 5 or 6, characterized in that The antibacterial agent is an oral medicament or an external application medicament.

8. The antibacterial agent according to claim 7, characterized in that The antibacterial agent is used for promoting delayed healing of wounds caused by or accompanying diabetes.

9. The antibacterial agent according to claim 7 or 8, characterized in that The external application agent further comprises a carrier and an acceptable auxiliary material, wherein the carrier is chitosan, and the auxiliary material is an aqueous solution of a polyoxyethylene-polyoxypropylene ether triblock copolymer.

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