Amphiphilic nigrin molecules, methods of making, and hypoglycemic and antibacterial applications for diabetic wounds

Abstract

The application provides an amphiphilic nobeliatin molecule, a preparation method thereof and a blood glucose lowering and antibacterial application to a diabetic wound. The application relates to the technical field of biomedical materials. The application synthesizes a novel amphiphilic nobeliatin molecule by taking 1-deoxynobeliatin as a hydrophilic part and an alkyl chain as a hydrophobic part. The compound can self-assemble into a cluster sugar molecule in an aqueous solution, has good blood glucose lowering activity through the "multivalent effect" of the cluster sugar-glycosidase, has good antibacterial activity through the tertiary amine structure of the azasugar molecule and the long alkyl chain and the bacterial membrane, and realizes the compatibility of the blood glucose lowering effect and the antibacterial effect. The synthesis raw material of the nobeliatin molecule provided by the application is rich in source, low in price, simple in preparation process and convenient for industrial application. Therefore, the discovery of the compound is expected to provide a promising treatment scheme for the development of blood glucose lowering and antibacterial integrated materials and the treatment of diabetic wounds, and has a wide potential application field.

Description

Technical Field

[0001] This invention relates to the field of biomedical materials technology, specifically to an amphiphilic nojirimycin molecule, its preparation method, and its hypoglycemic and antibacterial application in diabetic wounds. Background Technology

[0002] Diabetes mellitus, characterized by high blood sugar, is a common disease affecting the endocrine and metabolic systems and is one of the top ten chronic diseases worldwide. With the rising incidence of type 2 diabetes mellitus (T2DM), several T2DM-related complications are becoming increasingly prominent, including fatty liver disease, cancer, dementia, sarcopenia and frailty, as well as other diseases affecting the lungs, heart, and intestines. Among these, chronic diabetic wounds are a complex and common complication, posing a significant challenge to global healthcare systems and often referred to as "the cancer of diabetes." These wounds are typically characterized by high blood sugar, bacterial infection, and persistent inflammation, all of which collectively hinder wound healing. Current research on hypoglycemic and antibacterial strategies for chronic, non-healing diabetic wounds has proposed the following approaches: loading insulin and metformin to achieve systemic hypoglycemia (Gao et al. Appl. Surf. Sci. 2022, 576, 151825; Yang et al. Adv. Mater. 2025, 37, 2419158; Tian et al. Chem. Eng. J. 2025, 506, 160179.); loading glucose oxidase to achieve local hypoglycemia (Li et al. Chem. Eng. J. 2024, 487, 150545; Xiang et al. Acta Biomater. 2024, 182, 245.); and loading antibiotics onto a carrier for antibacterial action (Xu et al. Carbohydr. Polym. 2024, 324, 121543; Lin et al.). al. Macromol. Biosci. 2023, 23, 2300145; Yan et al. Int. J. Biol. Macromol. 2023, 281, 136166.), etc.

[0003] Nojirimycin is a natural hypoglycemic molecule found in mulberry leaves. Researchers have reported that its derivatives have good inhibitory activity against glucosidase, as shown in patent applications CN 119039209 A and CN 109456254 A. Additionally, there are reports on the antibacterial applications of nojirimycin derivatives (Esposito et al. ACS Pharmacol. Transl. Sci. 2024, 7, 1807.). However, no reports have yet documented a single material possessing both hypoglycemic and antibacterial functions. Summary of the Invention

[0004] The purpose of this invention is to provide an amphiphilic nojirimycin molecule, its preparation method, and its hypoglycemic and antibacterial application in diabetic wounds. This amphiphilic nojirimycin molecule has dual effects of hypoglycemic and antibacterial properties, and has great application prospects for wound healing in diabetic patients.

[0005] This invention is implemented as follows:

[0006] In a first aspect, the present invention provides an amphiphilic nojirimycin molecule, the structure of which is shown in formula (I):

[0007]

[0008] This invention designs and synthesizes an amphiphilic nojirimycin-like molecule. The introduction of azahesaccharide groups endows the material with biocompatibility and biological functions. Among them, the multivalent glycosidase inhibitor exhibits highly efficient and selective inhibitory activity against glycosidases. Furthermore, the long alkyl chain, as an intercalation molecule in bacterial membranes, possesses potential bacterial membrane targeting capabilities. In addition, the tertiary amine structure in the azahesaccharide acts as a molecule contributing to the negative charge of bacterial membranes, possessing potential bacterial adhesion and bacterial membrane targeting capabilities. This invention investigates the glycosidase inhibitory activity and antibacterial effects of the synthesized compound and applies it to wound healing in a diabetic mouse model, laying the foundation for the development of hypoglycemic and antibacterial compatible materials.

[0009] The amphiphilic nojirimycin-like molecule provided by this invention exhibits excellent in vitro α-glucosidase inhibitory activity based on the "multivalent effect" mechanism of glyco-glucosidase recognition. Experimental results show that the in vitro α-glucosidase (mouse-derived) inhibitory activity of AP-DNJ is K. i =0.09±0.003μM. Furthermore, 1-deoxynojirimycin is a nitrogenous sugar molecule with a tertiary amine structure, which can interact with the negative charge of the bacterial membrane; in addition, the long alkyl chain can effectively intercalate into the bacterial membrane, possessing potential antibacterial activity. In vitro antibacterial experiments demonstrated that AP-DNJ exhibits good antibacterial activity against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, drug-resistant Staphylococcus aureus, and drug-resistant Pseudomonas aeruginosa, exhibiting broad-spectrum antibacterial characteristics.

[0010] At 20 μM, AP-DNJ achieves a bactericidal rate of over 90% against common Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli, especially against Staphylococcus aureus, where the bactericidal rate approaches 98%. At 40 μM, AP-DNJ achieves a bactericidal rate of over 95% against drug-resistant bacteria, including drug-resistant Staphylococcus aureus and drug-resistant Pseudomonas aeruginosa.

[0011] In a diabetic mouse wound model infected with Staphylococcus aureus, AP-DNJ showed good hypoglycemic and antibacterial effects, and significantly promoted wound healing in diabetic mice after 12 days, demonstrating broad application prospects in the treatment of bacterial-infected diabetic mouse wounds.

[0012] Secondly, the present invention also provides a method for preparing the above-mentioned amphiphilic nojirimycin molecules, comprising the following steps:

[0013] S1. 1-Deoxynojirimycin and bromoazidotriethylene glycol (represented by M-1) were dissolved in N,N-dimethylformamide (DMF) to undergo a substitution reaction. DMF was then removed by vacuum distillation. Pyridine and acetic anhydride were added, and the hydroxyl groups on 1-deoxynojirimycin were protected by acetyl groups. Pyridine was distilled off, and intermediate M-2 was obtained by column chromatography.

[0014]

[0015] S2. Triynebutyric acid derivative (represented by M-3) and N,N-di(octadecyl)-1,2-ethylenediamine (represented by M-4) were added to DMF solvent and condensed under an inert atmosphere using 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU) and N,N-diisopropylethylamine (DIPEA) as catalysts. The intermediate M-5 was then obtained by column chromatography.

[0016]

[0017] S3. Intermediates M-2 and M-5 were dissolved in tetrahydrofuran (THF) solvent. Using sodium ascorbate and copper sulfate dissolved in aqueous solution as catalysts, the acetyl-protected nojirimycin-modified amphiphilic key intermediate AP-AcDNJ was prepared by click reaction. The key intermediate AP-AcDNJ was separated by column chromatography.

[0018]

[0019] S4. The key intermediate AP-AcDNJ was dissolved in anhydrous methanol, sodium methoxide was added, and the acetyl group was removed under alkaline conditions. Then, it was neutralized with hydrochloric acid, dialyzed, and freeze-dried to obtain the amphiphilic nojirimycin molecule.

[0020] The specific synthesis route is shown below:

[0021] (1) Synthesis of intermediates M-2 and M-5

[0022]

[0023] (2) Synthesis of target compound

[0024]

[0025] Compared with existing technologies, the method for preparing amphiphilic nojirimycin molecules provided by this invention has a short synthesis step, simple operation, mild reaction conditions, and high product purity, which has good industrial utilization value and high application value.

[0026] Furthermore, in S4, dialysis is performed using an aqueous dialysis bag with a molecular weight cutoff of 1000 for 24 hours.

[0027] Furthermore, in S4, freeze drying specifically involves: first freezing in a -80°C refrigerator, and then freeze-drying in a freeze dryer.

[0028] Further, in S1, the eluent for column chromatography separation is dichloromethane and methanol in a volume ratio of 20:1.

[0029] Furthermore, in S2, the eluent for column chromatography separation is dichloromethane and ethyl acetate in a volume ratio of 2:1.

[0030] Furthermore, in S3, the eluent for column chromatography separation is dichloromethane and methanol in a volume ratio of 20:1.

[0031] Furthermore, in S3, the reaction temperature of the click reaction is 55°C, and the atmosphere is a nitrogen atmosphere.

[0032] Furthermore, all of the above column chromatography separations used silica gel columns.

[0033] It should be noted that the inert atmosphere in this invention can be provided by conventional inert gases in the art, such as nitrogen and argon.

[0034] Furthermore, the amount of solvent used in this invention and the ratio between the reaction raw materials can be adjusted by those skilled in the art through conventional experiments, and this invention does not impose any special limitations.

[0035] Thirdly, the present invention provides the application of the above-mentioned amphiphilic nojirimycin molecules in the preparation of hypoglycemic and antibacterial drugs.

[0036] Furthermore, hypoglycemic drugs are drugs that reduce type 2 diabetes, and antibacterial drugs are drugs that inhibit the activity of Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, drug-resistant Staphylococcus aureus, and drug-resistant Pseudomonas aeruginosa.

[0037] The amphiphilic nojirimycin-like molecule provided by this invention exhibits significant in vitro inhibitory activity against α-glucosidase, as well as in vitro antibacterial activity against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, drug-resistant Staphylococcus aureus, and drug-resistant Pseudomonas aeruginosa. In vivo experiments have also confirmed these conclusions. Furthermore, the raw materials for the synthesis of this compound are abundant and inexpensive, and the preparation process is simple, facilitating industrial application. Therefore, the discovery of this compound holds promise for providing a highly promising therapeutic approach for both hypoglycemic and antibacterial purposes, with broad potential applications.

[0038] Furthermore, using diabetic mice infected with Staphylococcus aureus as a model, in vivo experiments were conducted. This invention demonstrated excellent in vivo hypoglycemic and antibacterial effects, effectively promoting wound healing and providing new possibilities for the development of integrated hypoglycemic and antibacterial materials. Attached Figure Description

[0039] Figure 1 To observe the antibacterial activity of the AP-DNJ compound prepared in Example 1 by plate coating method.

[0040] Figure 2 The image shows SEM images of the compound AP-DNJ prepared in Example 1 after interaction with different bacteria.

[0041] Figure 3 The graph shows the changes in postprandial blood glucose levels over time in diabetic mouse models after administration of the compounds AP-DNJ and Miglitol (a hypoglycemic molecule) prepared in Example 1 to the wounds of the mice.

[0042] Figure 4 To investigate the antibacterial activity of AP-DNJ (25 μM), AP-DNJ (50 μM), miglitol (a hypoglycemic molecule), and benzalkonium chloride (CAB, an antibacterial agent) at different time points in the wounds of diabetic mice infected with Staphylococcus aureus.

[0043] Figure 5 To investigate the wound healing of diabetic mice infected with Staphylococcus aureus after administration of AP-DNJ (25 μM), AP-DNJ (50 μM), Miglitol (a hypoglycemic molecule), and benzalkonium chloride (CAB, an antibacterial agent) at different time points. Detailed Implementation

[0044] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0045] Example 1: Preparation method of AP-DNJ

[0046] This embodiment provides a method for preparing amphiphilic nojirimycin molecules, comprising the following steps:

[0047] (1) Preparation of intermediate M-2:

[0048] 1-Deoxynojirimycin (321.9 mg, 2.0 mmol) was added to a 10 mL round-bottom flask, followed by 6 mL of DMF for ultrasonic dissolution. Anhydrous potassium carbonate (272.2 mg, 2.0 mmol) was added with stirring, followed by bromoazidotriethylene glycol M-1 (935.0 mg, 4.0 mmol). The reaction was carried out at 95 °C for 24 h, and the reaction was monitored by TLC. Post-treatment: First, the DMF solvent was distilled off under reduced pressure, and the product was dried under vacuum to obtain the crude product, which was not purified. Further, the crude product was dissolved in 6 mL of anhydrous pyridine, and DMAP (48.2 mg, 0.4 mmol) was added as a catalyst. Under nitrogen protection, the mixture was stirred in an ice bath for 20 min, and then acetic anhydride (1.1 mL, 6.0 mmol) was added dropwise using a constant-pressure dropping funnel. The reaction was carried out at room temperature for 5 h, and the reaction was monitored by TLC. Post-processing: Most of the pyridine was distilled off using a rotary evaporator, then dissolved in 50 mL of dichloromethane, washed three times with 1 M HCl, concentrated in dichloromethane, separated by column chromatography (petroleum ether: ethyl acetate = 1:1, volume ratio), and dried in a vacuum drying oven to obtain a viscous yellow liquid intermediate M-2.

[0049]

[0050] The characterization results of this compound are as follows:

[0051] M-2: 1 H NMR (600MHz, CDCl3): δ (ppm) 5.02-4.94 (m, 2H), 4.93-4.86 (m, 1H), 4.23 (dd, J = 12.9 ,2.3Hz,1H),4.13(dd,J=12.9,3.2Hz,1H),3.60(p,J=5.6Hz,3H),3.58-3.54(m,3H), 3.53(dd,J=6.4,3.8Hz,2H),3.48-3.43(m,J=10.1,5.0Hz,1H),3.33(t,J=5.0Hz,2H ),3.16(dd,J=11.6,5.2Hz,1H),2.93-2.89(m,J=14.8,7.3,4.5Hz,1H),2.86-2.82(m 1H),2.78-2.67(m,J=6.4,3.6,3.2Hz,1H),2.51-2.45(m,1H),2.01(s,3H),1.95(d,J=1.4Hz,6H),1.94(s,3H);

[0052] 13 C NMR (150MHz, CDCl3): δ (ppm) 170.94, 170.36, 170.00, 169.71, 74.69, 71.12-69.88 (m), 69.42 (d, J = 2.5 Hz), 68.40, 61.40, 59.57, 53.45, 50.86, 50.67,22.07 - 20.33 (m);

[0053] MS(ESI):C 20 H 32 N4O 10 H + ,m / z:489.2185[M+H] + .

[0054] (2) Preparation of intermediate M-5:

[0055] Triynylbutyric acid derivative M-3 (335 mg, 1.0 mmol) and N,N-di(octadecyl)-1,2-ethylenediamine M-4 (556 mg, 1.0 mmol) were added to DMF solvent. Under an inert atmosphere, the reaction was carried out at room temperature for 12 hours using 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU, 380 mg, 1.0 mmol) and N,N-diisopropylethylamine (DIPEA, 129 mg, 1.0 mmol) as catalysts. The reaction was monitored by TLC. Post-treatment: DMF was removed by vacuum distillation. The sample was dissolved in dichloromethane, washed three times with 1M HCl solution, concentrated, and separated by column chromatography (dichloromethane:ethyl acetate = 2:1, v / v). The solution was dried in a vacuum oven to obtain a white solid intermediate, M-5.

[0056]

[0057] The characterization results of this compound are as follows:

[0058] M-5: 1 H NMR (400MHz, CDCl3): δ (ppm) 6.03 (s, 1H), 4.18 (d, J = 2.4Hz, 6H), 3.87 (s,

[0059] 6H),3.40(s,2H),2.54(d,J=3.2Hz,8H),2.47(t,J=2.4Hz,3H),1.52(s,4H),1.29(s,62H),0.91(t,J=6.7Hz,6H);

[0060] 13 C NMR (100MHz, CDCl3): δ (ppm) 77.41, 77.09, 76.77, 74.70, 68.56, 59.32, 58.73, 53. 76,32.31,31.99,31.62,29.78,29.73,29.69,29.54,29.43,27.39,22.76,14.20;

[0061] FTMS(ESI):C 55 H 99 N3O5H + ,m / z 882.7654[M+H] + .

[0062] (3) Preparation of the key intermediate AP-AcDNJ:

[0063] Weigh M-2 (310.0 mg, 0.6 mmol) and M-5 (155.0 mg, 0.2 mmol) into a 100 mL round-bottom flask, add tetrahydrofuran solution, and purge the air from the flask using a vacuum pump. Weigh L-ascorbic acid sodium (125.8 mg, 0.6 mmol) and copper sulfate pentahydrate (158.5 mg, 0.6 mmol), dissolve them in aqueous solution (as catalyst), and then add them to the above reaction solution. Stir at 55 °C under a nitrogen atmosphere and react for 12 h. Monitor the reaction by thin-layer chromatography (TLC). Post-processing: Dihydrofuran was evaporated under reduced pressure using a rotary evaporator, then extracted with dichloromethane, washed once with water, and separated. The dichloromethane phase was concentrated using a rotary evaporator and separated by column chromatography (dichloromethane:methanol = 20:1, v / v) to prepare AP-AcDNJ.

[0064]

[0065] The characterization results of this compound are as follows:

[0066] AP-AcDNJ: 1H NMR (400MHz, CDCl3): δ (ppm) 7.78 (s, 3H), 6.65 (s, 1H), 5.13-4.91 (m, 9H), 4.57 (d, J = 6.2Hz, 12H), 4.33-4.15(m,6H),3.90(t,J=5.3Hz,6H),3.79(s,5H),3.66-3.45(m,20H),3.25(dd,J=11.5,5.1H z,5H),3.11(d,J=10.0Hz,4H),2.94(t,J=5.4Hz,6H),2.83-2.79(m,J=5.6,2.8Hz,3H),2.59-2.40 (m,9H),2.13-1.96(m,36H),1.67(d,J=8.7Hz,4H),1.26(d,J=2.4Hz,61H),0.89(t,J=6.8Hz,6H).

[0067] (4) Preparation of the target compound AP-DNJ:

[0068] AP-AcDNJ (115.0 mg, 0.1 mmol) and sodium methoxide (47.7 mg, 0.9 mmol) were weighed into a 100 mL round-bottom flask and dissolved in anhydrous methanol. The mixture was stirred at room temperature for 4 h, and the reaction was monitored by thin-layer chromatography (TLC). Post-treatment: The sodium methoxide was first neutralized with dilute hydrochloric acid, then water was added, and the mixture was transferred to a dialysis bag and dialyzed for 24 h, changing the water every 2 hours. Finally, the mixture was freeze-dried to obtain the final target compound, AP-DNJ.

[0069]

[0070] The characterization results of this compound are as follows:

[0071] AP-DNJ: 1H NMR (600MHz, DMSO-d6): δ (ppm) 8.00 (s, 3H), 4.71 (s, 1H), 4.50 (t, J = 5.3Hz, 6H), 4.47 (s, 6H), 3.81 (t, J = 5.3Hz ,6H),3.73(dd,J=11.7,2.3Hz,3H),3.63(s,6H),3.55(dd,J=11.8,3.6Hz,4H),3.50(dd,J=5.8,3.6Hz,7H),3.2 0-3.18(m,3H),3.09-3.00(m,5H),2.92-2.86(m,6H),2.86(dd,J=11.2,4.8Hz,3H),2.44-2.27(m,8H),2.22(t, J=7.6Hz,2H),2.07(t,J=10.8Hz,3H),2.00(m,3H),1.33(t,J=7.1Hz,5H),1.22(s,66H),0.84(t,J=7.0Hz,6H);

[0072] 13 C NMR (151MHz, DMSO-d6): δ (ppm) 143.86, 124.04, 78.89, 70.41, 69.52, 69.47, 69.10, 68.69, 68.08, 67.91, 6 6.60,64.09,58.81,57.71,53.42,51.13,49.30,31.26,28.99,28.94,28.81,28.67,26.61,22.04,13.82;

[0073] HRMS(ESI):C 91 H 172 N 15 O 23 H + ,m / z 1843.2692[M+H] + .

[0074] Example 2: Inhibitory activity of α-glucosidase in vitro

[0075] To evaluate the inhibitory effect of AP-DNJ synthesized in Example 1 on α-glucosidase activity in vitro, α-glucosidase was extracted from mouse intestines, with miglitol (a clinical hypoglycemic drug) used as a control group. Miglitol is a small molecule drug commonly used to treat diabetes. The glycosidase inhibitory activity of AP-DNJ was evaluated by comparing it with that of the synthesized AP-DNJ. The inhibitory activities of AP-DNJ and Miglitol on α-glucosidase (mouse-derived) are shown in Table 1.

[0076] Table 1. Inhibitory activities of AP-DNJ and Miglitol on α-glucosidase (mouse-derived) (K i (μM)

[0077] Glycosidase Miglitol AP-DNJ α-glucosidase (mice) 1.46±0.23 0.09±0.003

[0078] Table 1 shows that the inhibitory activity of AP-DNJ on α-glucosidase is K. i The concentration was 0.09 ± 0.003 μM, while the inhibitory activity of Miglitol against α-glucosidase was K. i The concentration was 1.46 ± 0.23 μM. This is consistent with the fact that AP-DNJ contains three nojirimycin molecules, and its activity is 5.4 times higher than that of miglitol. This indicates that AP-DNJ has excellent in vitro glycosidase inhibitory activity, laying the foundation for further research on the in vivo hypoglycemic effect of AP-DNJ.

[0079] Example 3: In vitro antibacterial activity of AP-DNJ

[0080] The bactericidal efficacy of compound AP-DNJ against five bacteria—Staphylococcus aureus (SA), Pseudomonas aeruginosa (PA), Escherichia coli, methicillin-resistant Staphylococcus aureus (MRSA), and multidrug-resistant Pseudomonas aeruginosa (MRD-PA)—was quantitatively evaluated using a 96-well plate assay. The bacterial survival rates after interaction with bacteria at different concentrations of compound AP-DNJ are shown in Table 2.

[0081] Table 2. Bacterial survival rate (%) after interaction with AP-DNJ.

[0082] concentration SA PA E. coli MRSA MRD-PA 2.5μM 60.9±3.5 72.2±1.7 76.0±1.1 92.0±0.4 92.6±0.8 5μM 34.9±2.1 42.3±2.2 44.7±1.1 63.7±0.4 70.6±1.4 10μM 19.7±1.0 26.2±1.1 28.4±1.1 46.3±1.0 54.2±0.7 20μM 2.0±0.4 6.0±1.6 10.9±1.1 26.7±0.6 33.3±1.8 40μM 1.2±0.1 1.1±0.6 2.1±0.1 4.3±0.7 4.7±0.7

[0083] As shown in Table 2, the bacterial survival rate gradually decreased with increasing concentration of compound AP-DNJ, and the antibacterial activity against bacteria showed a significant concentration dependence. When the concentration of AP-DNJ reached 20 μM, its bactericidal rate against Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli reached 90%, while the bactericidal rate against drug-resistant Staphylococcus aureus and multidrug-resistant Pseudomonas aeruginosa reached nearly 70%. At 40 μM, AP-DNJ achieved a bactericidal rate of 98% against Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli, while the bactericidal rate against drug-resistant Staphylococcus aureus and multidrug-resistant Pseudomonas aeruginosa exceeded 95%.

[0084] The antibacterial activity of AP-DNJ was observed by the plate coating method, and the results are as follows: Figure 1As shown, compared with the blank control group, AP-DNJ has a broad-spectrum antibacterial effect against different bacteria (Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, drug-resistant Staphylococcus aureus and drug-resistant Pseudomonas aeruginosa).

[0085] Example 4: Study on the antibacterial mechanism of AP-DNJ

[0086] The AP-DNJ prepared in Example 1 was reacted with five bacteria: Staphylococcus aureus (S. aureus or SA), Pseudomonas aeruginosa (P. aeruginosa or PA), Escherichia coli (E. coli), methicillin-resistant Staphylococcus aureus (MRSA), and multidrug-resistant Pseudomonas aeruginosa (MRD-PA). The morphological changes of the bacteria were then observed using scanning electron microscopy (SEM). The results are as follows: Figure 2 As shown. Figure 2 In the control group, the bacterial cells were plump and three-dimensional with very smooth surfaces. However, after treatment with AP-DNJ, the morphology of the bacteria underwent significant changes, specifically: cell membrane wrinkling, surface collapse, and leakage of contents. This indicates that AP-DNJ achieves its antibacterial effect by disrupting the bacterial cell membrane.

[0087] Example 5: Study on the in vivo hypoglycemic activity of AP-DNJ

[0088] Based on the good glycosidase inhibitory activity of AP-DNJ in vitro, this invention investigated the in vivo hypoglycemic effect of AP-DNJ by administering it via wound site. Blood samples were collected from the tail vein to measure postprandial blood glucose levels in diabetic mice. The administration method was as follows: mice were anesthetized with the gaseous anesthetic isoflurane, and different drugs and dosages were administered at the wound site.

[0089] The in vivo hypoglycemic effect of AP-DNJ was evaluated using a glucose tolerance test and compared with that of Miglitol. The effects of Miglitol and AP-DNJ on postprandial blood glucose levels in mice are shown in Table 3. Figure 3 As shown.

[0090] Table 3. In vivo glucose-lowering activity of AP-DNJ and Miglitol

[0091]

[0092] From Table 3 and Figure 3It was found that, when the postprandial blood glucose level of mice was measured, the blood glucose level was at its peak. Two hours after administration, the blood glucose level dropped to its lowest point: the control group decreased from 606.2±10.9 mg / dL to 452.9±14.5 mg / dL, the Miglitol (150 μM) group decreased from 605.3±10.2 mg / dL to 367.2±15.3 mg / dL, the AP-DNJ (25 μM) group decreased from 602.6±11.2 mg / dL to 342.0±13.5 mg / dL, and the AP-DNJ (50 μM) group decreased from 601.2±2.7 mg / dL to 325.8±8.5 mg / dL. Subsequently, the blood glucose level gradually and slowly increased. Compared with the control group, AP-DNJ and Miglitol had similar in vivo hypoglycemic activities. At doses of 25 μM and 50 μM, AP-DNJ resulted in significantly lower blood glucose levels at 15, 30, 60, and 120 minutes compared to the miglitol group. The miglitol doses were 6 and 3 times higher than those of AP-DNJ, respectively. AP-DNJ exhibited a favorable multivalent effect.

[0093] Therefore, AP-DNJ can reduce postprandial blood glucose levels in mice in a short time when administered through the wound site, which is better than the clinical hypoglycemic drug Miglitol, providing a new approach for the treatment of chronic and difficult-to-heal diabetic wounds.

[0094] Example 6: In vivo antibacterial activity study

[0095] Diabetic mice infected with Staphylococcus aureus were constructed by administering benzalkonium chloride (BAC), miglitol, AP-DNJ (25 μM), and AP-DNJ (50 μM) via wound site administration. Bacterial activity at the mouse wound site was observed at different time points using the plate spread method. Results are shown below. Figure 4 .

[0096] Depend on Figure 4 It was observed that the number of bacteria at the wound site decreased significantly with the duration of treatment. By day eight, bacteria were essentially eliminated from the wounds in both the benzalkonium chloride (BAC) group and the AP-DNJ group, especially in the AP-DNJ (50 μM) group. The effect of the AP-DNJ (25 μM) group was lower than that of the AP-DNJ (50 μM) group, showing a concentration-dependent effect.

[0097] Example 7: Study on Diabetic Wound Healing

[0098] Diabetic mice infected with Staphylococcus aureus were constructed by administering benzalkonium chloride (BAC), miglitol, AP-DNJ (25 μM), and AP-DNJ (50 μM) via wound site treatment. Wound healing was observed at different time points, and the results are shown in Table 4. Figure 5 .

[0099] Table 4. Wound non-healing rate at different times

[0100]

[0101] From Table 4 and Figure 5 It was observed that, with prolonged treatment, the wound healing effect in the control group was not significant, and the wounds remained reddish. On day 3, the wound non-healing rate was 81.5% in the control group, 37.3% in the benzalkonium chloride group (BAC), 42.8% in the miglitol group, 39.9% in the AP-DNJ (25μM) group, and 39.0% in the AP-DNJ (50μM) group. The AP-DNJ group showed a similar wound healing rate to the benzalkonium chloride group. On day 8, the wound non-healing rate of AP-DNJ (50μM) reached 11.2%, equivalent to four times that of the control group. On day 12, the wound healing rate of the AP-DNJ-treated group was nearly 98%, better than the 95% in the benzalkonium chloride group. The experimental results indicate that AP-DNJ has a good effect on promoting wound healing.

[0102] This embodiment uses diabetic mice infected with Staphylococcus aureus as a model to measure the effect of AP-DNJ on wound healing by administering it to the wound. AP-DNJ can effectively promote the healing of diabetic wounds, providing a new idea for the development of materials that integrate hypoglycemic and antibacterial properties.

[0103] In summary, AP-DNJ not only possesses good α-glucosidase inhibitory activity and broad-spectrum antibacterial activity, but also exhibits good in vivo hypoglycemic activity and bactericidal effect, and significantly promotes the healing of diabetic wounds.

Claims

1. An amphiphilic wild-type nigrin molecule, characterized in that, The structure is shown as formula (I): Formula (I).

2. The method of preparing the amphipathic wild-type nigericin molecule according to claim 1, characterized in that, The method comprises the following steps: S1, 1-deoxynojirimycin is dissolved with bromoazide triethylene glycol in DMF to have a substitution reaction, and then DMF is removed by distillation under reduced pressure; pyridine and acetic anhydride are added, and then pyridine is distilled off, and an intermediate M-2 is obtained by column chromatography separation; M-2 S2, the trialkynyl butyric acid derivative is added to N , N - Di(octadecyl)-1,2-ethanediamine is added to DMF solvent, and condensation reaction is carried out under an inert atmosphere with 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N-diisopropylethylamine as catalysts, and then the intermediate M-5 is obtained by column chromatography. M-5 S3, the intermediate M-2 is dissolved with the intermediate M-5 in a tetrahydrofuran solvent, and an aqueous solution of sodium ascorbate and copper sulfate is added, and after click reaction, column chromatography separation is performed to obtain a key intermediate AP-AcDNJ; AP-AcDNJ S4, the key intermediate AP-AcDNJ is dissolved in anhydrous methanol solvent, sodium methoxide is added, and the acetyl group is removed under alkaline conditions, and then neutralized with hydrochloric acid, then dialyzed, and freeze-dried to obtain an amphiphilic wild mycin molecule.

3. The method of preparing an amphipathic wild-type nigericin molecule according to claim 2, characterized in that, In step S4, the water phase dialysis bag with a molecular weight cut-off of 1000 is used for dialysis, and the dialysis time is 24 hours.

4. The method for preparing amphiphilic nojirimycin-like molecules according to claim 2, characterized in that, In step S4, the freeze-drying is specifically: first frozen in a-80℃ refrigerator, and then freeze-dried by a freeze dryer.

5. The method for preparing the amphiphilic nojirimycin-like molecule according to claim 2, characterized in that, In step S1, the eluent for column chromatography separation is petroleum ether and ethyl acetate in a volume ratio of 1:

1.

6. The method for preparing amphiphilic nojirimycin-like molecules according to claim 2, characterized in that, In step S2, the eluent for column chromatography separation is dichloromethane and ethyl acetate in a volume ratio of 2:

1.

7. The method for preparing the amphiphilic nojirimycin-like molecule according to claim 2, characterized in that, In step S3, the eluent for column chromatography separation is dichloromethane and methanol in a volume ratio of 20:

1.

8. The method for preparing the amphiphilic nojirimycin-like molecule according to claim 2, characterized in that, In step S3, the reaction temperature of click reaction is 55℃, and the atmosphere is nitrogen atmosphere.

9. The use of the amphiphilic wild mycin molecule of claim 1 or the amphiphilic wild mycin molecule prepared by the method of any one of claims 2-8 in the preparation of a hypoglycemic and antibacterial drug; the antibacterial drug refers to a drug against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, drug-resistant Staphylococcus aureus or multi-drug resistant Pseudomonas aeruginosa.

10. A medicament capable of promoting wound healing in a diabetic mouse, characterized in that, The use of the amphiphilic wild mycin molecule of claim 1 or the amphiphilic wild mycin molecule prepared by the method of any one of claims 2-8.

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