Amphiphilic nojirimycin molecule, preparation method thereof and hypoglycemic and antibacterial application of amphiphilic nojirimycin molecule to diabetic wounds

Abstract

The invention provides an amphipathic nojirimycin molecule, a preparation method thereof and application of the amphipathic nojirimycin molecule in reducing blood glucose and resisting bacteria on diabetic wounds. The invention relates to the technical field of biomedical materials. According to the invention, 1-deoxynojirimycin is taken as a hydrophilic part, an alkyl chain is taken as a hydrophobic part, and a novel amphiphilic nojirimycin molecule is synthesized. The compound can be self-assembled into a cluster sugar molecule in an aqueous solution, and has good hypoglycemic activity through a'multivalent effect 'of cluster sugar-glycosidase; through the action of a tertiary amine structure of an azasugar molecule and a long alkyl chain with a bacterial membrane, the azasugar has good antibacterial activity, and compatibility of a hypoglycemic effect and an antibacterial effect is realized. The nojirimycin molecule provided by the invention is rich in synthesis raw material 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 development of blood glucose reducing and antibacterial integrated materials and treatment of diabetic wounds, and the compound has a wide potential application field.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedical materials, in particular to an amphiphilic nojirimycin molecule and a preparation method thereof, and its application in reducing blood sugar and providing antibacterial treatment for diabetic wounds. Background Art

[0002] Diabetes, characterized by high blood sugar levels, is a common disease that affects the endocrine and metabolic systems and is one of the top ten chronic diseases worldwide. With the increasing incidence of type 2 diabetes mellitus (T2DM), some complications associated with T2DM have become increasingly prominent, including fatty liver disease, cancer, dementia, sarcopenia, and frailty, as well as other diseases of the lungs, heart, and intestines. Among them, chronic diabetic wounds are a complex and common complication and a major challenge facing the global healthcare system, earning them the nickname "the cancer of diabetes." Such wounds are typically characterized by high blood sugar levels, bacterial infection, and persistent inflammation, which together hinder wound healing. At present, the following strategies have been proposed for the research on hypoglycemic and antibacterial effects of chronic and difficult-to-heal diabetic wounds: loading insulin and metformin to achieve systemic hypoglycemic effect (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 on the wound surface for local hypoglycemic effect (Li et al. Chem. Eng. J. 2024, 487, 150545; Xiang et al. Acta Biomater. 2024, 182, 245.); loading antibiotics on the carrier for antibacterial effect (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 discovered 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. Furthermore, there are reports of nojirimycin derivatives for antibacterial applications (Esposito et al. ACS Pharmacol. Transl. Sci. 2024, 7, 1807). However, a single material combining both hypoglycemic and antibacterial properties has yet to be reported. Summary of the Invention

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

[0005] The present invention is achieved in that:

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

[0007]

[0008] The present invention designs and synthesizes an amphiphilic nojirimycin-like molecule. The introduction of azasugar groups gives the material biocompatibility and biological functionality. Among them, the multivalent glycosidase inhibitor has high inhibitory activity and selectivity against glycosidases. On the other hand, the long alkyl chain, as an embedded molecule in the bacterial membrane, has potential bacterial membrane targeting ability. In addition, the tertiary amine structure in the azasugar, as a molecule that acts on the negative charge of the bacterial membrane, has potential bacterial adhesion ability and bacterial membrane targeting ability. The present invention studies the glycosidase inhibitory activity and antibacterial effect of the synthesized compound, and applies it to wound healing in a diabetic mouse model, laying the foundation for the development of compatible materials with hypoglycemic and antibacterial properties.

[0009] The amphiphilic nojirimycin molecules provided by the present invention have excellent in vitro α-glucosidase inhibitory activity based on the "multivalent effect" mechanism of sugar-glycosidase recognition. The experimental results show that the in vitro α-glucosidase (mouse source) inhibitory activity of AP-DNJ is K i = 0.09 ± 0.003 μM. 1-Deoxynojirimycin is an azasugar molecule with a tertiary amine structure that can interact with the negative charge of bacterial membranes. Furthermore, the long alkyl chain effectively embeds into bacterial membranes, demonstrating potential antibacterial activity. In vitro antibacterial studies have demonstrated that AP-DNJ exhibits strong antibacterial activity against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, and drug-resistant Staphylococcus aureus and Pseudomonas aeruginosa, demonstrating broad-spectrum antibacterial properties.

[0010] At 20 μM, AP-DNJ achieved a bactericidal rate of over 90% against common Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli, with a particularly high bactericidal rate of nearly 98% against Staphylococcus aureus. At 40 μM, AP-DNJ achieved a bactericidal rate of over 95% against drug-resistant bacteria, including drug-resistant Staphylococcus aureus and Pseudomonas aeruginosa.

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

[0012] In a second aspect, the present invention further provides a method for preparing the above-mentioned amphiphilic nojirimycin-like molecules, comprising the following steps:

[0013] S1. Dissolve 1-deoxynojirimycin and bromoazidotriethylene glycol (represented by M-1) in N,N-dimethylformamide (DMF) to undergo a substitution reaction, and then remove DMF by distillation under reduced pressure; then add pyridine and acetic anhydride to protect the hydroxyl group on 1-deoxynojirimycin with an acetyl group, distill off the pyridine, and separate by column chromatography to obtain the intermediate M-2;

[0014]

[0015] S2. A trialkynylbutyric acid derivative (represented by M-3) and N,N-dioctadecyl-1,2-ethylenediamine (represented by M-4) are added to a DMF solvent, and a condensation reaction is carried out under an inert atmosphere using 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) and N,N-diisopropylethylamine (DIPEA) as catalysts, followed by column chromatography to obtain an intermediate M-5.

[0016]

[0017] S3. Intermediates M-2 and M-5 are dissolved in tetrahydrofuran (THF) solvent, and sodium ascorbate and copper sulfate are dissolved in aqueous solution as catalysts to prepare acetyl-protected nojirimycin-modified amphiphilic key intermediate AP-AcDNJ through a click reaction; the key intermediate AP-AcDNJ is separated by column chromatography;

[0018]

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

[0020] The specific synthetic route is as follows:

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

[0022]

[0023] (2) Synthesis of target compound

[0024]

[0025] Compared with the prior art, the preparation method of the amphiphilic nojirimycin-like molecules provided by the present invention has short synthesis steps, simple operation, mild reaction conditions, high purity of the obtained product, good industrial utilization value, and high promotion and application value.

[0026] Furthermore, in S4, dialysis is performed using an aqueous dialysis bag with a molecular weight cut-off of 1000, and the dialysis time is 24 hours.

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

[0028] Furthermore, 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 was 55° C. and the atmosphere was nitrogen.

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

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

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

[0035] In a third aspect, the present invention provides the use of the above-mentioned amphiphilic nojirimycin-like molecules in the preparation of hypoglycemic and antibacterial drugs.

[0036] Furthermore, the hypoglycemic drug is a drug that reduces type II diabetes, and the antibacterial drug is a drug that inhibits the activity of Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, drug-resistant Staphylococcus aureus and drug-resistant Pseudomonas aeruginosa.

[0037] The amphiphilic nojirimycin-like molecules provided by the present invention have significant in vitro inhibitory activity against α-glucosidase, and in vitro antibacterial activity against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, drug-resistant Staphylococcus aureus, and drug-resistant Pseudomonas aeruginosa. In vivo experiments have confirmed the above conclusions. Furthermore, the synthetic raw materials of this compound are abundant and inexpensive, and the preparation process is simple, making it easy for industrial application. Therefore, the discovery of this compound is expected to provide a promising treatment option for hypoglycemic and antibacterial treatment, with a wide range of potential applications.

[0038] Furthermore, in vivo experiments were conducted using diabetic mice infected with Staphylococcus aureus as a model. The present invention demonstrated excellent hypoglycemic and antibacterial effects in vivo, effectively promoted wound healing, and provided new possibilities for the development of integrated hypoglycemic and antibacterial materials. BRIEF DESCRIPTION OF THE DRAWINGS

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

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

[0041] Figure 3 This is a graph showing changes in postprandial blood glucose levels over time in diabetic mouse models after the compound AP-DNJ prepared in Example 1 and Miglitol (a hypoglycemic molecule) were administered to the wounds of the mice.

[0042] Figure 4 The antibacterial activities of AP-DNJ (25 μM), AP-DNJ (50 μM), Miglitol (a single hypoglycemic molecule), and benzalkonium chloride (CAB, an antibacterial agent) were evaluated at different times in the wounds of diabetic mice infected with Staphylococcus aureus.

[0043] Figure 5 To investigate the healing of wounds in diabetic mice infected with Staphylococcus aureus, we administered AP-DNJ (25 μM), AP-DNJ (50 μM), Miglitol (a single hypoglycemic molecule), and Benzalkonium Chloride (CAB, an antibacterial agent) at different times. DETAILED DESCRIPTION

[0044] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0045] Example 1: Preparation of AP-DNJ

[0046] This embodiment provides a method for preparing an amphiphilic nojirimycin-like molecule, 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, 6 mL of DMF was added and ultrasonically dissolved, anhydrous potassium carbonate (272.2 mg, 2.0 mmol) was added under stirring, and then bromoazidotriethylene glycol M-1 (935.0 mg, 4.0 mmol) was added. The mixture was reacted at 95 ° C for 24 h, and the reaction was monitored by TLC. Post-treatment: First, vacuum distillation was performed to distill off the DMF solvent and vacuum dried to obtain a crude product without separation and purification. Further, 6 mL of anhydrous pyridine was added to dissolve the crude product, and the catalyst DMAP (48.2 mg, 0.4 mmol) was added. After stirring for 20 min in an ice bath, acetic anhydride (1.1 mL, 6.0 mmol) was added dropwise using a constant pressure dropping funnel. The reaction was reacted at room temperature for 5 h and the reaction was monitored by TLC. Post-treatment: Most of the pyridine was evaporated 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 the 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] Trialkynylbutyric acid derivative M-3 (335 mg, 1.0 mmol) and N,N-dioctadecyl-1,2-ethylenediamine M-4 (556 mg, 1.0 mmol) were added to DMF solvent. Under an inert atmosphere, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU, 380 mg, 1.0 mmol) and N,N-diisopropylethylamine (DIPEA, 129 mg, 1.0 mmol) were used as catalysts. The reaction was allowed to proceed at room temperature for 12 hours and monitored by TLC. Post-treatment: DMF was removed by distillation under reduced pressure. The sample was dissolved in dichloromethane and washed three times with 1M HCl solution. After concentration, column chromatography (dichloromethane:ethyl acetate = 2:1, volume ratio) was performed and dried in a vacuum oven to obtain intermediate M-5 as a white solid.

[0056]

[0057] The characterization results of the 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 key intermediate AP-AcDNJ:

[0063] M-2 (310.0 mg, 0.6 mmol) and M-5 (155.0 mg, 0.2 mmol) were weighed into a 100 mL round-bottom flask, and the tetrahydrofuran solution was added. The air in the flask was evacuated using a vacuum pump. Sodium L-ascorbate (125.8 mg, 0.6 mmol) and copper sulfate pentahydrate (158.5 mg, 0.6 mmol) were weighed, dissolved in aqueous solution (as a catalyst), and then added to the reaction solution. The mixture was stirred at 55°C under a nitrogen atmosphere for 12 h and monitored by thin-layer chromatography (TLC). Post-treatment: The tetrahydrofuran was removed by rotary evaporation under reduced pressure, and then extracted with dichloromethane. The mixture was then washed once with water and separated. The dichloromethane phase was concentrated by rotary evaporation and separated by column chromatography (dichloromethane:methanol = 20:1, volume ratio) to prepare AP-AcDNJ.

[0064]

[0065] The characterization results of the 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 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 absolute anhydrous methanol. The mixture was stirred at room temperature for 4 hours and monitored by thin-layer chromatography (TLC). Post-treatment: The sodium methoxide was first neutralized with dilute hydrochloric acid. Water was then added and the mixture was transferred to a dialysis bag and dialyzed for 24 hours, with the water changed every 2 hours. Finally, the product was freeze-dried in a freeze dryer to obtain the target compound, AP-DNJ.

[0069]

[0070] The characterization results of the 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 in vitro, α-glucosidase was extracted from the mouse intestine, and miglitol (a clinical hypoglycemic drug) was used as a control group. Miglitol, a small molecule drug commonly used to treat diabetes, was compared with the synthesized AP-DNJ to evaluate its glycosidase inhibitory activity. The inhibitory activities of AP-DNJ and Miglitol against α-glucosidase (mouse source) are shown in Table 1.

[0076] Table 1 Inhibitory activity of AP-DNJ and Miglitol on α-glucosidase (mouse origin) (K i , μM)

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

[0078] As shown in Table 1, the inhibitory activity K of AP-DNJ on α-glucosidase i was 0.09±0.003μM, while the inhibitory activity of Miglitol on α-glucosidase K i The results were 1.46 ± 0.23 μM. This is consistent with the fact that AP-DNJ contains three nojirimycin molecules, which increases its activity by 5.4-fold compared to miglitol. This indicates that AP-DNJ has excellent in vitro glycosidase inhibitory activity and lays the foundation for further research on the hypoglycemic effect of AP-DNJ in vivo.

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

[0080] A 96-well plate assay was used to quantitatively evaluate the bactericidal efficacy of compound AP-DNJ against five bacterial species: Staphylococcus aureus (SA), Pseudomonas aeruginosa (PA), Escherichia coli (E. coli), methicillin-resistant S. aureus (MRSA), and multidrug-resistant P. aeruginosa (MRD-PA). Table 2 shows the bacterial survival rates after exposure to different concentrations of compound AP-DNJ.

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

[0082] concentration SA PA E. coli MRSA MRD-PA 2.5 μM 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 10 μM 19.7±1.0 26.2±1.1 28.4±1.1 46.3±1.0 54.2±0.7 20 μM 2.0±0.4 6.0±1.6 10.9±1.1 26.7±0.6 33.3±1.8 40 μM 1.2±0.1 1.1±0.6 2.1±0.1 4.3±0.7 4.7±0.7

[0083] As can be seen from Table 2, with the increase of the concentration of compound AP-DNJ, the survival rate of bacteria gradually decreased, and the antibacterial activity against bacteria showed obvious 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, the bactericidal rate of AP-DNJ against Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli reached 98%, while the bactericidal rate against drug-resistant Staphylococcus aureus and multidrug-resistant Pseudomonas aeruginosa reached more than 95%.

[0084] The antibacterial activity of AP-DNJ was observed by plate coating method. Figure 1As shown in the figure, compared with the blank control group (control), 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 respectively reacted with Staphylococcus aureus (S. aureus or SA), Pseudomonas aeruginosa (P. aeruginosa or PA), Escherichia coli (E. coli), drug-resistant Staphylococcus aureus (MRSA) and multidrug-resistant Pseudomonas aeruginosa (MRD-PA), and the morphological changes of the bacteria were observed by scanning electron microscopy (SEM). The results are shown in FIG. Figure 2 shown. Figure 2 In the blank control group, the bacterial cells were plump and three-dimensional, with a very smooth surface. However, after exposure to AP-DNJ, the bacterial morphology underwent significant changes, manifested by wrinkles in the cell membrane, surface collapse, and leakage of contents. This suggests that AP-DNJ achieves its antibacterial effect by disrupting the bacterial biofilm.

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

[0088] Based on AP-DNJ's demonstrated robust glycosidase inhibitory activity in vitro, the present invention investigated the in vivo hypoglycemic effect of AP-DNJ via wound administration. Blood samples were collected via the tail vein to measure postprandial blood glucose levels in diabetic mice. The administration method involved administering different drugs at varying doses to the wound site while the mice were anesthetized with the gaseous anesthetic isoflurane.

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

[0090] Table 3 In vivo hypoglycemic activity of AP-DNJ and Miglitol

[0091]

[0092] From Table 3 and Figure 3It can be seen that the postprandial blood glucose level of mice was first measured, and the blood glucose value was at its peak. 2 hours after administration, the blood glucose level dropped to the lowest: the blank group dropped from 606.2±10.9 mg / dL to 452.9±14.5 mg / dL, the Miglitol (150μM) group dropped from 605.3±10.2 mg / dL to 367.2±15.3 mg / dL, the AP-DNJ (25μM) group dropped from 602.6±11.2 mg / dL to 342.0±13.5 mg / dL, and the AP-DNJ (50μM) group dropped from 601.2±2.7 mg / dL to 325.8±8.5 mg / dL. Then the blood glucose level gradually increased slowly. Compared with the blank group, AP-DNJ and Miglitol have similar hypoglycemic activity in vivo. At doses of 25 μM and 50 μM, AP-DNJ significantly lowered blood glucose levels at 15, 30, 60, and 120 minutes compared to the miglitol group. The miglitol dose was six and three times higher than that of AP-DNJ, respectively. AP-DNJ demonstrated a favorable multivalent effect.

[0093] Therefore, AP-DNJ can reduce the postprandial blood glucose level of mice in a short period of time by administering it to the wound, which is better than the clinical hypoglycemic drug Miglitol, and provides ideas 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 and the following drugs were administered to the wounds of mice: benzalkonium chloride (BAC), miglitol (Miglitol), AP-DNJ (25μM) and AP-DNJ (50μM). The bacterial status of the wounds of mice at different time points was observed by plate coating method. The results are shown in the figure. Figure 4 .

[0096] Depend on Figure 4 As can be seen, the number of bacteria in the wound decreased significantly over the course of treatment. By day eight, bacteria were essentially eliminated from the wounds in both the benzalkonium chloride (BAC) and AP-DNJ groups, particularly in the AP-DNJ (50 μM) group. The effect in the AP-DNJ (25 μM) group was lower than that in the AP-DNJ (50 μM) group, demonstrating a concentration-dependent effect.

[0097] Example 7: Diabetic Wound Healing Study

[0098] Diabetic mice infected with Staphylococcus aureus were constructed and the following drugs were administered to the wounds of mice: benzalkonium chloride (BAC), miglitol, AP-DNJ (25μM) and AP-DNJ (50μM). The wound healing of mice was observed at different time points. The results are shown in Table 4 and Figure 5 .

[0099] Table 4 Wound unhealing rate at different time

[0100]

[0101] From Table 4 and Figure 5 It can be seen that as time goes on, the wound healing effect of the blank group mice is not obvious, and the wound appears red. On the 3rd day, the wound healing rate of the blank group (Control) was 81.5%, the wound healing rate of the benzalkonium chloride group (BAC) was 37.3%, the wound healing rate of the miglitol group was 42.8%, the wound healing rate of AP-DNJ (25μM) was 39.9%, and the wound healing rate of AP-DNJ (50μM) was 39.0%. The AP-DNJ group had a wound healing rate similar to that of the benzalkonium chloride group. On the 8th day, the wound healing rate of AP-DNJ (50μM) reached 11.2%, which is 4 times that of the blank group. On the 12th day, it can be visually observed that the healing of the wounds treated with AP-DNJ was nearly 98%, which is better than the 95% of the benzalkonium chloride group. The experimental results show that AP-DNJ has a good effect in promoting wound healing.

[0102] In this example, diabetic mice infected with Staphylococcus aureus were used as a model to determine the effect of AP-DNJ on wound healing by wound administration. AP-DNJ can effectively promote diabetic wound healing, providing new ideas for the development of integrated hypoglycemic and antibacterial materials.

[0103] In summary, AP-DNJ not only has good α-glucosidase inhibitory activity and broad-spectrum antibacterial activity, but also has good in vivo hypoglycemic activity and bactericidal effect, and has a significant effect on promoting the healing of diabetic wounds.

Claims

1. An amphiphilic nojirimycin-like molecule, characterized in that: Its structure is shown in formula (I):

2. The method for preparing the amphiphilic nojirimycin-like molecule according to claim 1, characterized in that: The steps include: S1. Dissolving 1-deoxynojirimycin and bromoazidotriethylene glycol in DMF to cause a substitution reaction, and then removing DMF by distillation under reduced pressure; then adding pyridine and acetic anhydride, and then distilling off the pyridine, and separating by column chromatography to obtain the intermediate M-2; S2. Add a trialkynylbutyric acid derivative and N,N-dioctadecyl-1,2-ethylenediamine to a DMF solvent, and carry out a condensation reaction under an inert atmosphere using 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N-diisopropylethylamine as catalysts, followed by column chromatography to obtain an intermediate M-5; S3, dissolving intermediates M-2 and M-5 in tetrahydrofuran solvent, adding aqueous solutions of sodium ascorbate and copper sulfate, performing a click reaction, and separating by column chromatography to obtain the key intermediate AP-AcDNJ; S4. The key intermediate AP-AcDNJ was dissolved in anhydrous methanol solvent, sodium methoxide was added, and the acetyl group was removed under alkaline conditions. The product was then neutralized with hydrochloric acid, dialyzed, and freeze-dried to obtain the amphiphilic nojirimycin-like molecule.

3. The method for preparing the amphiphilic nojirimycin-like molecule according to claim 2, wherein: In step S4, dialysis is performed using an aqueous dialysis bag with a molecular weight cut-off of 1000, and the dialysis time is 24 hours.

4. The method for preparing the amphiphilic nojirimycin-like molecule according to claim 2, wherein: In step S4, freeze drying is specifically as follows: first freezing in a -80°C refrigerator, and then freeze drying in a freeze dryer.

5. The method for preparing the amphiphilic nojirimycin-like molecule according to claim 2, wherein: 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 the amphiphilic nojirimycin-like molecule according to claim 2, wherein: 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, wherein: 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, wherein: In step S3, the click reaction temperature is 55° C. and the atmosphere is nitrogen.

9. Use of the amphiphilic nojirimycin molecule according to claim 1 or the amphiphilic nojirimycin molecule prepared by the method according to any one of claims 2 to 8 in the preparation of hypoglycemic and antibacterial drugs.

10. A drug capable of promoting wound healing in diabetic mice, characterized by: The amphiphilic nojirimycin molecule comprises the amphiphilic nojirimycin molecule according to claim 1 or the amphiphilic nojirimycin molecule prepared by the method according to any one of claims 2 to 8.

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