Fluorescent phenanthroimidazole-amine copolymers, methods of making and antimicrobial applications

Abstract

The application discloses a series of fluorescent phenanthroimidazole-amine copolymers, a preparation method thereof and antibacterial application. The series of compounds are converted into N,N-dimethyl-4-(1H-phenanthro[9,10-d]imidazol-2-yl)aniline with fluorescent properties by taking phenanthrenequinone as a mother body, then the N,N-dimethyl-4-(1H-phenanthro[9,10-d]imidazol-2-yl)aniline is connected with various different aliphatic amines and heterocyclic amines through different length alkyl connectors as lipophilic groups, and a series of fluorescent phenanthroimidazole-amine copolymers are synthesized. The phenanthroimidazole-amine copolymers prepared by the application show strong bacteriostatic effect on staphylococcus aureus and various methicillin-resistant staphylococcus aureus (MRSA), good water solubility and excellent biological safety, and have high yield, simple synthesis method, easy availability and suitability for large-scale production, and are expected to be further developed into semi-synthetic antibacterial drugs in clinical application.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry technology, specifically relating to phenanthrimidazole-amine copolymers, their preparation methods, and antibacterial applications. Background Technology

[0002] Globally, antibiotic resistance is rapidly becoming an urgent public health problem (Hobson C, Chan AN, Wright G D. The antibiotic resistome: a guide for the discovery of natural products as antimicrobial agents[J]. Chemical Reviews, 2021, 121(6):3464-3494). Among the many drug-resistant bacteria, methicillin-resistant Staphylococcus aureus (MRSA) is the most well-known, listed by the World Health Organization (WHO) as one of the twelve priority pathogens posing a serious threat to human health (Ahmed SA, Baris E, Go DS, et al. Assessing the global economic and poverty effects of antimicrobial resistance[J]. World Bank Policy Research Working Paper, 2017(8133)). MRSA not only exhibits significant resistance to β-lactam antibiotics, but also resistance to a variety of antibiotics, including fluoroquinolones, macrolides, and tetracyclines. Recent studies have even shown that MRSA exhibits high resistance to antibiotics such as vancomycin, which serve as a last line of defense (Cong Y, Yang S, Rao X. Vancomycin-resistant Staphylococcus aureus infections: A review of case updating and clinical features[J]. Journal of advanced research, 2020, 21: 169-176).

[0003] Antimicrobial peptides (AMPs) have attracted considerable attention due to their broad-spectrum antimicrobial activity. They exert their effects by disrupting the stability of microbial membranes, thereby reducing bacterial resistance. However, the therapeutic application of AMPs is limited by their stability, toxicity, and production costs (Lazzaro BP, Zasloff M, Rolf J. Antimicrobial peptides: Application informed by evolution[J]. Science, 2020, 368(6490): eaau5480). Therefore, the design of AMP mimics that mimic the functional properties of natural AMPs while overcoming their limitations has become a promising solution. Imidazole derivatives possess broad antibacterial activity, including some commercially available products such as metronidazole, tinidazole, econazole, and clotrimazole (Zhang L, Peng XM, Damu GLV, et al. Comprehensive review in current developments of imidazole-based medicinal chemistry[J]. Medicinal research reviews, 2014, 34(2):340-437). Among them, benzimidazole and dibenzimidazole are important antibacterial core structures and have been used to develop new antibacterial agents. However, there is still a need to find new compounds to address antibiotic resistance and realize the possibility of integrated diagnosis and treatment. Summary of the Invention

[0004] Objective of the Invention: This invention aims to at least solve one of the aforementioned technical problems existing in the prior art. To this end, this invention provides a method for preparing and applying a phenanthrimidazole-amine copolymer, which exhibits excellent antibacterial activity against Staphylococcus aureus ATCC 29213 and various clinically isolated Gram-positive bacteria such as MRSA, and solves problems related to biosafety and poor water solubility.

[0005] Technical Solution: To achieve the above-mentioned objectives, the technical solution adopted by this invention is as follows:

[0006] A fluorescent phenanthreneimidazole-amine copolymer or a pharmaceutically acceptable salt thereof, the copolymer having the structural formula shown in formula (III):

[0007]

[0008] Where n = 3, 4, or 5, and R is selected from... or R1 and R2 are independently selected from hydrogen or C1-C6 alkyl groups; A is selected from C, N, S or O.

[0009] Preferably, n = 3, 4, or 5, and R is selected from...

[0010] Preferably, the copolymer is selected from the following compounds:

[0011]

[0012] Preferably, the pharmaceutically acceptable salt is selected from the following compounds:

[0013]

[0014] Wherein, n and R are as described in the claims.

[0015] The present invention also provides a method for preparing the phenanthrimidazole-amine copolymer, comprising the following steps:

[0016] (1) Under alkaline conditions, bromoacetyl bromide is reacted with secondary amine R-NH to obtain intermediate 1;

[0017] (2) Under alkaline conditions, intermediate 1 continues to react with dimethylamine to generate small molecule peptide fragment 2;

[0018] (3) Using phenanthrenequinone as a substrate, it was reacted with p-diethylaminobenzaldehyde and ammonium acetate in a three-component Debus-Radziszewski imidazole synthesis reaction to obtain fluorescent N,N-dimethyl-4-(1H-phenanthrene[9,10-d]imidazol-2-yl)aniline, which is intermediate I;

[0019] (4) Under alkaline conditions, intermediate I is reacted with dibromoalkane to synthesize intermediate II;

[0020] (5) Finally, intermediate II is reacted with intermediate 2 under alkaline conditions to generate a series of phenanthrimidazole-amine copolymers:

[0021]

[0022] Wherein, n and R are as described in any one of claims 1-4.

[0023] Preferably, in step (1), the base in the alkaline conditions is K2CO3, the reaction solvent is selected from dichloromethane solution, the reaction temperature is 0±2℃, and the reaction time is 5h-12h;

[0024] Preferably, in step (2), the base in the alkaline conditions is K2CO3, the reaction solvent is selected from acetone solution, the reaction temperature is 95-100℃, and the reaction time is 1h-2h.

[0025] Preferably, in step (3), the reaction temperature is 100±2℃, the reaction time is 1h-2h, and the reaction solvent is glacial acetic acid solution;

[0026] Preferably, in step (4), the base in the alkaline conditions is K2CO3, the molar ratio of intermediate I to the base is 1:4-1:5, the molar ratio of intermediate I to dibromoalkane is 1:4-1:8, the reaction temperature is 45-60℃, the reaction time is 12h-14h, and the reaction solvent is anhydrous acetonitrile.

[0027] Preferably, in step (5), the base in the alkaline conditions is K2CO3, the reaction molar ratio of intermediate II to the base is 1:4-1:5, the reaction molar ratio of intermediate II to intermediate 2 is 1:4-1:5, the reaction temperature is 70-80℃, the reaction time is 12h-16h, and the reaction solvent is ethanol.

[0028] The present invention also provides the use of the phenanthrimidazole-amine copolymer or a pharmaceutically acceptable salt thereof in the preparation of antibacterial drugs.

[0029] Furthermore, the antibacterial drug can inhibit Staphylococcus aureus or methicillin-resistant Staphylococcus aureus.

[0030] Finally, this invention provides the application of the described phenanthrimidazole-amine copolymer or its pharmaceutically acceptable salt in the preparation of therapeutic antibacterial agents. The phenanthrimidazole-amine copolymer of this invention can dynamically exhibit a bactericidal process, achieving therapeutic integration.

[0031] Technical Effects: The phenanthrimidazole-amine copolymer designed based on the structural and functional properties of antimicrobial peptides in this invention exhibits excellent antibacterial activity against Staphylococcus aureus and clinically isolated methicillin-resistant Staphylococcus aureus (MRSA). Furthermore, the introduction of tertiary amine fragments significantly enhances the interaction between the compound and bacteria, thereby enhancing the fluorescence signal and potentially enabling a dynamic bactericidal process, achieving integrated diagnosis and treatment. The copolymer of this invention possesses low hemolysis rate, high water solubility, and stability, while also exhibiting good biocompatibility and in vivo safety, demonstrating promising development prospects and potential for translational applications. Attached Figure Description

[0032] Figure 1 The dynamic bactericidal curve of compound III30 is shown.

[0033] Figure 2 The fluorescence properties of compound III30.

[0034] Figure 3 The in vivo blood routine and blood biochemical indices of compound III30.

[0035] Figure 4 The change in bacterial load on mouse skin for compound III30.

[0036] Figure 5 The image shows the 1H-NMR spectrum of compound III30.

[0037] Figure 6 The image shows the 13C-NMR spectrum of compound III30. Detailed Implementation

[0038] The technical solutions in the embodiments of the present invention will be clearly and completely described below, but the implementation of the present invention is not limited thereto.

[0039] Unless otherwise specified, the reagents, methods and equipment used in this invention are all conventional reagents, methods and equipment in this technical field.

[0040] Example 1: Preparation of intermediate compound 1A-J

[0041] A mixture of amine R-NH (1.0 mmol) and K₂CO₃ (1.5 mmol) was dissolved in CH₂Cl₂ in a pear-shaped flask and stirred for 30 min under N₂ protection and an ice bath. Bromoacetyl bromide (1.5 mmol) was slowly added dropwise to the mixture at 0 °C and stirred for 5 h. The reaction was monitored by thin-layer chromatography (TLC) until completion, then extracted with EtOAc, dried over anhydrous Na₂SO₄, and concentrated to give intermediate 1A-J. The yield was 89.2%.

[0042] The R group in the above amine is selected from

[0043] The products obtained are 1A-1J in sequence.

[0044] Example 2 Preparation of intermediate 2A-J

[0045] Dimethylamine aqueous solution (1.5 mmol) and K2CO3 (1.5 mmol) were added to an acetone solution of intermediate 1A-J. The mixture was stirred at room temperature for 12-18 h, then extracted, concentrated, and separated by column chromatography to obtain intermediate 2A-J (related groups and serial numbers are the same as in Example 1). The yield was 87.2%.

[0046] Example 3 Preparation of Intermediate I

[0047] In a 50 mL round-bottom flask, phenanthrenequinone (150 mg, 0.72 mmol, 1 equiv) and p-diethylaminobenzaldehyde (172.1 mg, 0.72 mmol, 1 equiv) were dissolved in glacial acetic acid, and then an appropriate amount of ammonium acetate (1.11 g, 14.4 mmol, 20 equiv) was added. The mixture was stirred at 100 °C for 1 h, and the reaction solution was cooled to room temperature and poured into ice water. The solid was collected by filtration and washed with ice water. The solid was then dried under vacuum to give intermediate I in 73.3% yield.

[0048] Example 4 Preparation of compound II1-3

[0049] K₂CO₃ was added to an anhydrous MeCN solution of compound I (1.0 mmol) and the corresponding brominated alkane (7.0 mmol). The reaction was stirred at 75 °C for 12–14 h, and intermediates II₁–3 were obtained by extraction, concentration, and column chromatography in yields of 47.0–58.7%.

[0050] Example 5 Preparation of Compound III1-30

[0051] Intermediate II1-3 (1 mmol) and intermediate 2A-J (3 mmol) were weighed into a pressure flask, dissolved in an appropriate amount of anhydrous ethanol, and stirred under reflux at 78 °C. Thin-layer chromatography (TLC) was used to detect the reaction until completion. After the reaction was completed, the target compound III1-30 was separated by thin-layer chromatography (PTLC) with a yield of 21.4-95.1%.

[0052] The target compounds III1-30 are the specific compounds III1-III30 listed in the technical solution. The final solid products obtained in Examples 6-35 are the products in the form of the bromide salts of the above compounds III1-III30.

[0053] Example 6 Compound III1

[0054] Compound III1 was synthesized using the method described in Example 5. The physicochemical properties of compound III1 are as follows:

[0055] 1) White solid;

[0056] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (400MHz) characteristics:

[0057] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 70.1%. 1H NMR(600MHz DMSO-d6)δ:8.98(d,J=8.4Hz,1H,-Ph),8.86(d,J=8.4Hz,1H,-Ph),8.58-8.59(m,1H,-Ph),8.49(d,J= 5.4Hz,1H,-NH-),8.38(d,J=7.8Hz,1H,-Ph),7.77-7.79(m,1H,-Ph),7.62-7.72(m,5H,-Ph),6.90(d, J=8.4Hz,2H,-Ph),4.70(t,J=7.2Hz,2H,-CH2-),3.96(s,2H,-CH2-),3.58-3.61(m,2H,-CH2-),3.12( s,6H,N-CH3),3.00-3.05(m,8H,N-CH3,-CH2-),2.25-2.30(m,2H,-CH2-),0.93(t,J=7.2Hz,3H,-CH3); 13 C NMR(150MHz DMSO-d6)δ:162.4,152.9,150.7,137.1,130.5,127.9,127.46,127.4,127.1,126,7,125.3,125.1,124.7,124. 4,123.4,122.6,121.8,120.7,116.9,111.7,61.3,60.7,51.2,45.1,43.3,39.8,33.4,23.6,13.9,HRMS(ESI)C 32 H 38 BrN5O[M-Br] + calcd=508.3071; found=508.3075.

[0058] Example 7 Compound III2

[0059] Compound III2 was synthesized using the method described in Example 5. The physicochemical properties of compound III2 are as follows:

[0060] 1) White solid;

[0061] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (400MHz) characteristics

[0062] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 70.1%. 1H NMR(400MHz DMSO-d6)δ:8.97(d,J=8.0Hz,1H,-Ph),8.85(d,J=8.4Hz,1H,-Ph),8.58(d,J=7.6Hz,1H,-Ph),8.50(s,1H,-N H-),8.38(d,J=8.0Hz,1H,-Ph),7.77-7.80(m,1H,-Ph),7.61-7.73(m,5H,-Ph),6.90(d,J=8.8Hz,2H,-Ph),4 .70(t,J=7.2Hz,2H,-CH2-),4.00(s,2H,-CH2-),3.58-3.62(m,2H,-CH2-),3.12(s,6H,N-CH3),3.03(s,6H,N -CH3),2.92-2.97(m,2H,-CH2-),2.28(s,2H,-CH2-),1.29-1.34(m,2H,-CH2-),0.76(t,J=7.6Hz,3H,-CH3); 13 CNMR(100MHz DMSO-d6)δ:162.7,152.9,150.7,137.2,130.6,128.0,127.54,127.5,127.1,126.8,125.4,125.2,124.8,1 24.5,123.4,122.7,121.9,120.7,117.0,111.7,61.4,60.8,51.3,43.4,40.3,23.7,21.8,11.2,HRMS(ESI)C 33 H 40 BrN5O[M-Br] + calcd=522.3227; found=522.3231.

[0063] Example 8 Compound III3

[0064] Compound III3 was synthesized using the method described in Example 5. The physicochemical properties of compound III3 are as follows:

[0065] 1) White solid;

[0066] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (400MHz) characteristics:

[0067] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 46.1%. 1H NMR(400MHz DMSO-d6)δ:8.97(d,J=8.4Hz,1H,-Ph),8.85(d,J=8.4Hz,1H,-Ph),8.57(d,J=7.6Hz,1H,-Ph),8.49(s,1H,-N H-),8.38(d,J=8.4Hz,1H,-Ph),7.76-7.80(m,1H,-Ph),7.61-7.73(m,5H,-Ph),6.90(d,J=8.8Hz,2H,-Ph),4. 69(t,J=7.2Hz,2H,-CH2-),4.00(s,2H,-CH2-),3.59(d,J=7.6Hz,2H,-CH2-),3.12(s,6H,N-CH3),3.03(s,6H, N-CH3),2.94-2.99(m,2H,-CH2-),2.28(s,2H,-CH2-),1.16-1.32(m,6H,-CH2-),0.81(t,J=6.8Hz,3H,-CH3); 13 CNMR(100MHz DMSO-d6)δ:162.6,152.9,150.7,137.2,130.6,128.0,127.5,127.1,126.8,125.4,125.2,124.8,124.5,123. 4,122.7,121.9,120.8,117.0,111.7,61.4,60.8,51.3,43.4,38.4,28.4,28.1,23.7,21.6,13.8; HRMS(ESI)C 35 H 44 BrN5O[M-Br] + calcd=550.3540; found=550.3545.

[0068] Example 9 Compound III4

[0069] Compound III4 was synthesized using the method described in Example 5. The physicochemical properties of compound III4 are as follows:

[0070] 1) White solid;

[0071] 3) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (400MHz) characteristics:

[0072] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 82.3%. 1H NMR(400MHz DMSO-d6)δ:8.97(d,J=8.4Hz,1H,-Ph),8.85(d,J=8.4Hz,1H,-Ph),8.64(s,1H,-NH-),8.57(d,J=8.0Hz,1H,- Ph),8.38(d,J=8.4Hz,1H,-Ph),7.77(t,J=7.6Hz,1H,-Ph),7.61-7.73(m,5H,-Ph),6.90(d,J=8.4Hz,2H,-Ph) ,4.69(t,J=7.6Hz,2H,-CH2-),4.04(s,2H,-CH2-),3.60-3.64(m,2H,-CH2-),3.13(s,6H,N-CH3),3.03(s,6H, N-CH3),2.95-2.99(m,2H,-CH2-),2.29(s,2H,-CH2-),1.15-1.31(m,8H,-CH2-),0.80(t,J=7.2Hz,3H,-CH3); 13 CNMR(100MHz DMSO-d6)δ:162.7,152.9,150.7,137.2,130.6,128.0,127.5,127.1,126.8,125.4,125.2,124.8,124.5,123. 4,122.7,121.9,120.8,117.0,111.7,61.4,60.8,51.2,43.4,38.4,28.4,28.1,23.7,21.6,13.8; HRMS(ESI)C 36 H 46 BrN5O[M-Br] + calcd=564.3697; found=564.3703.

[0073] Example 10 Compound III5

[0074] Compound III5 was synthesized using the method described in Example 5. The physicochemical properties of compound III5 are as follows:

[0075] 1) White solid;

[0076] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (400MHz) characteristics:

[0077] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 40.8%. 1H NMR(400MHz DMSO-d6)δ:8.99(d,J=8.4Hz,1H,-Ph),8.87(d,J=8.4Hz,1H,-Ph),8.60(d,J=7.6Hz,1H,-Ph),8.3 8(d,J=8.0Hz,1H,-Ph),7.63-7.81(m,6H,-Ph),6.91(d,J=8.8Hz,2H,-Ph),4.72(t,J=7.2Hz,2H,- CH2-),4.42(s,2H,-CH2-),3.61-3.65(m,2H,-CH2-),3.56(s,2H,-CH2-),3.43(s,2H,-CH2-),3.1 3(s,6H,N-CH3),3.04(s,6H,N-CH3),2.53(s,2H,-CH2-),2.41(s,2H,-CH2-),2.25(s,2H,-CH2-); 13 C NMR(100MHz DMSO-d6)δ:161.8,154.4,150.8,137.0,130.8,128.1,127.6,127.58,127.5,127.2,125.7,125.6,125.1,1 25.0,124.6,123.5,121.9,120.9,117.7,111.7,66.0,60.5,59.5,51.7,46.8,43.8,26.3,23.3; HRMS(ESI)C 34 H 40 BrN5OS[M-Br]calcd=566.2948; found=566.2948.

[0078] Example 11 Compound III6

[0079] Compound III6 was synthesized using the method described in Example 5. The physicochemical properties of compound III6 are as follows:

[0080] 1) White solid;

[0081] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (400MHz) characteristics:

[0082] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 37.3%. 1H NMR(600MHz DMSO-d6)δ:8.99(d,J=8.4Hz,1H,-Ph),8.87(d,J=8.4Hz,1H,-Ph),8.60(d,J=7.8Hz,1H,-Ph),8.38(d,J= 8.4Hz,1H,-Ph),7.77-7.80(m,1H,-Ph),7.63-7.74(m,5H,-Ph),6.91(d,J=9.0Hz,2H,-Ph),4.73(t,J=6.6 Hz,2H,-CH2-),4.30(s,2H,-CH2-),3.62-3.65(m,2H,-CH2-),3.14(s,6H,N-CH3),3.06-3.08(m,4H,-CH2- ),3.04(s,6H,N-CH3),2.19-2.24(m,2H,-CH2-),0.87(d,J=2.4Hz,3H,-CH3),0.85(t,J=7.2Hz,3H,-CH3); 13 C NMR(150MHzDMSO-d6)δ:162.0,152.7,150.8,130.6,128.0,127.6,127.4,127.2,125.6,125.0,124.5 ,123.4,122.4,121.8,120.7,111.6,60.0,59.0,51.6,43.4,40.5,39.9,23.4,13.3,12.4; HRMS(ESI)C 34 H 42 BrN5O[M-Br] + calcd=536.3384; found=536.3391.

[0083] Example 12 Compound III7

[0084] Compound III7 was synthesized using the method described in Example 5. The physicochemical properties of compound III7 are as follows:

[0085] 1) White solid;

[0086] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (600MHz) characteristics:

[0087] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 47.7%. 1H NMR(400MHz DMSO-d6)δ:9.01(d,J=8.4Hz,1H,-Ph),8.90(d,J=8.4Hz,1H,-Ph),8.64(d,J=7.2Hz,1H,-Ph),8.41(d,J=8 .0Hz,1H,-Ph),7.65-7.83(m,6H,-Ph),6.93(d,J=8.8Hz,2H,-Ph),4.79(s,2H,-CH2-),4.30(s,2H,-CH2-) ,3.59(d,J=6.8Hz,2H,-CH2-),3.14(s,6H,N-CH3),3.05(s,6H,N-CH3),2.83-2.92(m,4H,-CH2-),2.18(d, J=16.4Hz,2H,-CH2-),1.19-1.23(m,4H,-CH2-),0.66(t,J=5.2Hz,3H,-CH3),0.62(t,J=7.6Hz,3H,-CH3); 13 C NMR(100MHz DMSO-d6)δ:162.5,154.3,151.2,130.9,128.3,127.9,127.7,127.5,127.4,127.3,125.4,124.9,124.6,124.0, 123.7,122.2,122.0,121.0,111.7,59.8,58.9,52.0,47.8,46.7,43.8,23.4,20.9,19.9,10.9,10.7; HRMS(ESI)C 36 H 46 BrN5O[M-Br] + calcd=564.3697; found=564.3699.

[0088] Example 13 Compound III8

[0089] Compound III8 was synthesized using the method described in Example 5. The physicochemical properties of compound III8 are as follows:

[0090] 1) White solid;

[0091] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (600MHz) characteristics:

[0092] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 57.5%, (400MHz DMSO-d6) δ: 8.97 (d, J = 8.4Hz, 1H, -Ph), 8.85 (d, J = 8.4Hz, 1H, -Ph), 8.57 (d, J = 7.6Hz, 1H, -Ph), 8.36 (d, J = 8.0Hz, 1H, -Ph), 7.74-7.78 (m, 1H, -Ph), 7.60-7.72 (m, 6H, -Ph), 6.90 (d, J = 8.4Hz, 2H, -Ph), 4.71 (t, J = 6.4Hz, 2H, -CH2) -),4.28(s,2H,-CH2-),3.60-3.64(m,2H,-CH2-),3.15(s,6H,N-CH3),3.03(s,6H,N-CH3),2.92(d,J=6.0Hz,2H, -CH2-),2.18(s,2H,-CH2-),1.08-1.25(m,8H,-CH2-),0.77(t,J=7.6Hz,3H,-CH3),0.73(t,J=6.8Hz,3H,-CH3); 13 C NMR(100MHz DMSO-d6)δ:162.4,153.1,150.7,137.3,130.7,128.0,127.6,127.4,127.1,126.7,125.4,125.2,124.8,124.5,123.5, 122.7,121.9,120.7,117.0,111.7,59.9,58.9,51.8,46.0,44.8,43.3,29.7,28.7,19.4,19.2,13.56,13.5,HRMS(ESI)C 38 H 50 BrN5O[M-Br] + calcd=592.4010; found=592.4020.

[0093] Example 14 Compound III9

[0094] Compound III9 was synthesized using the method described in Example 5. The physicochemical properties of compound III9 are as follows:

[0095] 1) White solid;

[0096] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (600MHz) characteristics:

[0097] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 57.5%, (400MHz DMSO-d6) δ: 8.97 (d, J = 8.4Hz, 1H, -Ph), 8.85 (d, J = 8.4Hz, 1H, -Ph), 8.57 (d, J = 7.6Hz, 1H, -Ph), 8.36 (d, J = 8.0Hz, 1H, -Ph), 7.74-7.78 (m, 1H, -Ph), 7.60-7.72 (m, 6H, -Ph), 6.90 (d, J = 8.4Hz, 2H, -Ph), 4.71 (t, J = 6.4Hz, 2H, -CH2) -),4.28(s,2H,-CH2-),3.60-3.64(m,2H,-CH2-),3.15(s,6H,N-CH3),3.03(s,6H,N-CH3),2.92(d,J=6.0Hz,2H, -CH2-),2.18(s,2H,-CH2-),1.08-1.25(m,8H,-CH2-),0.77(t,J=7.6Hz,3H,-CH3),0.73(t,J=6.8Hz,3H,-CH3); 13 C NMR(100MHz DMSO-d6)δ:162.4,153.1,150.7,137.3,130.7,128.0,127.6,127.4,127.1,126.7,125.4,125.2,124.8,124.5,123.5, 122.7,121.9,120.7,117.0,111.7,59.9,58.9,51.8,46.0,44.8,43.3,29.7,28.7,19.4,19.2,13.56,13.5,HRMS(ESI)C 38 H 50 BrN5O[M-Br] + calcd=592.4010; found=592.4020.

[0098] Example 15 Compound III10

[0099] Compound III10 was synthesized using the method described in Example 5. The physicochemical properties of compound III10 are as follows:

[0100] 1) White solid;

[0101] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (400MHz) characteristics:

[0102] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 39.2%. 1H NMR(600MHz DMSO-d6)δ:8.98(d,J=8.4Hz,1H,-Ph),8.86(d,J=8.4Hz,1H,-Ph),8.59(d,J=7.8Hz,1H,-Ph),8.37(d,J=8.4Hz,1H,-Ph) ,7.76-7.79(m,1H,-Ph),7.61-7.73(m,5H,-Ph),6.90(d,J=8.4Hz,2H,-Ph),4.72(t,J=7.2Hz,2H,-CH2-),4.34(s,2H,-C H2-),3.68-3.71(m,2H,-CH2-),3.15(s,6H,N-CH3),3.04(s,6H,N-CH3),2.95(d,J=7.2Hz,2H,-CH2-),2.89(d,J=7.8Hz, 2H,-CH2-),2.17-2.22(m,2H,-CH2-),1.71-1.82(m,2H,-CH2-),0.68(d,J=6.6Hz,3H,-CH3),0.66(d,J=6.6Hz,3H,-CH3); 13 C NMR(150MHz DMSO-d6)δ:163.3,152.7,150.7,130.5,128.0,127.5,127.4,127.1,125.4,125.1,124.8,124.5,123.4, 122.5,121.8,120.7,111.6,60.1,59.1,53.5,52.3,51.6,43.4,26.5,25.5,23.6,19.6,19.2; HRMS(ESI)C 38 H 50 BrN5O[M-Br] + calcd=592.4010; found=592.4017.

[0103] Example 16 Compound III11

[0104] Compound III11 was synthesized using the method described in Example 5. The physicochemical properties of compound III11 are as follows:

[0105] 1) Yellow solid;

[0106] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (400MHz) characteristics:

[0107] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 39.2%. 1H NMR(600MHz DMSO-d6)δ:8.98(d,J=8.4Hz,1H,-Ph),8.86(d,J=8.4Hz,1H,-Ph),8.59(d,J=7.8Hz,1H,-Ph),8.37(d,J=8.4Hz,1H,-Ph) ,7.76-7.79(m,1H,-Ph),7.61-7.73(m,5H,-Ph),6.90(d,J=8.4Hz,2H,-Ph),4.72(t,J=7.2Hz,2H,-CH2-),4.34(s,2H,-C H2-),3.68-3.71(m,2H,-CH2-),3.15(s,6H,N-CH3),3.04(s,6H,N-CH3),2.95(d,J=7.2Hz,2H,-CH2-),2.89(d,J=7.8Hz, 2H,-CH2-),2.17-2.22(m,2H,-CH2-),1.71-1.82(m,2H,-CH2-),0.68(d,J=6.6Hz,3H,-CH3),0.66(d,J=6.6Hz,3H,-CH3); 13 C NMR(150MHz DMSO-d6)δ:163.3,152.7,150.7,130.5,128.0,127.5,127.4,127.1,125.4,125.1,124.8,124.5,123.4, 122.5,121.8,120.7,111.6,60.1,59.1,53.5,52.3,51.6,43.4,26.5,25.5,23.6,19.6,19.2; HRMS(ESI)C 38 H 50 BrN5O[M-Br] + calcd=592.4010; found=592.4017.

[0108] Example 17 Compound III12

[0109] Compound III12 was synthesized using the method described in Example 5. The physicochemical properties of compound III12 are as follows:

[0110] 1) Yellow solid;

[0111] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (400MHz) characteristics:

[0112] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 63.9%, 1H NMR(600MHz DMSO-d6)δ:8.97(d,J=7.8Hz,1H,-Ph),8.85(d,J=9.0Hz,1H,-Ph),8.58-8.59(m,1H,-Ph),8.53(t,J=5.4Hz,1H,- NH-),8.38(d,J=7.8Hz,1H,-Ph),7.77-7.80(m,1H,-Ph),7.62-7.73(m,5H,-Ph),6.91(d,J=8.4Hz,2H,-Ph),4.71 (t,J=7.8Hz,2H,-CH2-),3.98(s,2H,-CH2-),3.41(s,2H,-CH2-),3.11(s,6H,N-CH3),3.01-3.04(m,8H,N-CH3,-C H2-),1.79-1.84(m,2H,-CH2-),1.67-1.73(m,2H,-CH2-),1.37-1.40(m,2H,-CH2-),0.80(t,J=7.8Hz,3H,-CH3); 13 C NMR(150MHz DMSO-d6)δ:162.7,153.0,150.7,136.9,130.5,127.9,127.4,127.3,127.1,126.6,125.3,125.2,124.7,124.4,1 23.4,122.7,121.8,120.7,117.1,111.6,63.6,61.8,51.0,45.8,40.2,39.9,26.7,21.8,19.1,11.2; HRMS(ESI)C 34 H 42 BrN5O[M-Br] + calcd=536.3384; found=536.3390.

[0113] Example 18 Compound III13

[0114] Compound III13 was synthesized using the method described in Example 5. The physicochemical properties of compound III13 are as follows:

[0115] 1) Yellow solid;

[0116] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (400MHz) characteristics:

[0117] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 63.9%, 1H NMR(600MHz DMSO-d6)δ:8.97(d,J=7.8Hz,1H,-Ph),8.85(d,J=9.0Hz,1H,-Ph),8.58-8.59(m,1H,-Ph),8.53(t,J=5.4Hz,1H,- NH-),8.38(d,J=7.8Hz,1H,-Ph),7.77-7.80(m,1H,-Ph),7.62-7.73(m,5H,-Ph),6.91(d,J=8.4Hz,2H,-Ph),4.71 (t,J=7.8Hz,2H,-CH2-),3.98(s,2H,-CH2-),3.41(s,2H,-CH2-),3.11(s,6H,N-CH3),3.01-3.04(m,8H,N-CH3,-C H2-),1.79-1.84(m,2H,-CH2-),1.67-1.73(m,2H,-CH2-),1.37-1.40(m,2H,-CH2-),0.80(t,J=7.8Hz,3H,-CH3); 13 C NMR(150MHz DMSO-d6)δ:162.7,153.0,150.7,136.9,130.5,127.9,127.4,127.3,127.1,126.6,125.3,125.2,124.7,124.4,1 23.4,122.7,121.8,120.7,117.1,111.6,63.6,61.8,51.0,45.8,40.2,39.9,26.7,21.8,19.1,11.2; HRMS(ESI)C 34 H 42 BrN5O[M-Br] + calcd=536.3384; found=536.3390.

[0118] Example 19 Compound III14

[0119] Compound III14 was synthesized using the method described in Example 5. The physicochemical properties of compound III14 are as follows:

[0120] 1) Brown solid;

[0121] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (600MHz) characteristics:

[0122] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 66.2%. 1H NMR(600MHz DMSO-d6)δ:8.97(d,J=7.8Hz,1H,-Ph),8.85(d,J=8.4Hz,1H,-Ph),8.58(s,1H,-Ph),8.57(s,1H,-NH-),8.38(d ,J=8.4Hz,1H,-Ph),7.77-7.80(m,1H,-Ph),7.62-7.69(m,5H,-Ph),6.90(d,J=9.0Hz,2H,-Ph),4.71(t,J=7.8H z,2H,-CH2-),3.98(s,2H,-CH2-),3.38-3.41(m,2H,-CH2-),3.11(s,6H,N-CH3),3.03-3.06(m,8H,N-CH3,-CH2 -),1.80-1.82(m,2H,-CH2-),1.71-1.72(m,2H,-CH2-),1.19-1.24(m,8H,-CH2-),0.80(t,J=6.6Hz,3H,-CH3); 13 C NMR(150MHzDMSO-d6)δ:162.6,153.0,150.6,137.0,130.5,127.9,127.4,127.2,127.0,126.7,125.3,125.2,124.6,124.4,123 .4,122.7,121.8,120.7,117.2,111.6,63.6,61.8,51.0,45.8,39.9,38.4,30.7,28.4,26.7,25.8,21.9,19.1,13.7; HRMS(ESI)C 37 H 48 BrN5O[M-Br] + calcd=578.3853; found=578.3860.

[0123] Example 20 Compound III15

[0124] Compound III15 was synthesized using the method described in Example 5. The physicochemical properties of compound III15 are as follows:

[0125] 1) White solid;

[0126] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (400MHz) characteristics:

[0127] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 51.6%. 1H NMR(600MHz DMSO-d6)δ:8.99(d,J=8.4Hz,1H,-Ph),8.87(d,J=8.4Hz,1H,-Ph),8.60(d,J=7.8Hz,1H,-Ph),8.40(d,J=7.8Hz,1H,-Ph) ,7.78-7.80(m,1H,-Ph),7.64-7.74(m,5H,-Ph),6.92(d,J=9.0Hz,2H,-Ph),4.73(t,J=7.2Hz,2H,-CH2-),4.42(s,2H,-CH 2-),3.68(d,J=4.8Hz,2H,-CH2-),3.60(d,J=4.2Hz,2H,-CH2-),3.46-3.49(m,2H,-CH2-),3.13(s,6H,N-CH3),3.04(s,6H ,N-CH3),2.66(d,J=4.2Hz,2H,-CH2-),2.55(d,J=4.2Hz,2H,-CH2-),1.79-1.83(m,2H,-CH2-),1.64-1.69(m,2H,-CH2-); 13 C NMR(150MHzDMSO-d6)δ:161.9,152.8,150.8,130.6,127.9,127.5,127.3,127.2,125.5,125.2,124.9,124.5 ,123.5,122.5,121.8,120.9,111.6,63.6,60.1,51.1,47.2,45.9,43.7,26.68,26.6,26.2,19.0; HRMS(ESI)C 35 H 42 BrN5OS[M-Br] + calcd=580.3105; found=580.3109.

[0128] Example 21 Compound III16

[0129] Compound III16 was synthesized using the method described in Example 5. The physicochemical properties of compound III16 are as follows:

[0130] 1) Yellow solid;

[0131] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (400MHz) characteristics:

[0132] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 71.7%. 1H NMR(600MHz DMSO-d6)δ:8.97(d,J=8.4Hz,1H,-Ph),8.85(d,J=8.4Hz,1H,-Ph),8.57(d,J=7.8Hz,1H,-Ph),8.38(d,J=8.4Hz ,1H,-Ph),7.76-7.78(m,1H,-Ph),7.62-7.72(m,5H,-Ph),6.90(d,J=8.4Hz,2H,-Ph),4.71(t,J=7.2Hz,2H,-CH2 -),4.33(s,2H,-CH2-),3.48-3.50(m,2H,-CH2-),3.20-3.24(m,4H,-CH2-),3.14(s,6H,N-CH3),3.03(s,6H,N-C H3),1.79(d,J=6.0Hz,2H,-CH2-),1.62(s,2H,-CH2-),1.06(t,J=7.2Hz,3H,-CH3),0.94(t,J=7.2Hz,3H,-CH3); 13 C NMR(150MHzDMSO-d6)δ:162.3,153.1,150.6,137.1,130.5,127.9,127.4,127.2,127.0,126.7,125.3,125.2,124.6,12 4.4,123.4,122.7,121.8,120.7,117.2,111.6,63.2,59.6,51.2,45.8,43.9,40.8,26.7,19.1,13.6,12.5; HRMS(ESI)C 35 H 44 BrN5O[M-Br] + calcd=550.3540; found=550.3547.

[0133] Example 22 Compound III17

[0134] Compound III17 was synthesized using the method described in Example 5. The physicochemical properties of compound III17 are as follows:

[0135] 1) Yellow solid;

[0136] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (400MHz) characteristics:

[0137] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 71.7%. 1H NMR(600MHz DMSO-d6)δ:8.97(d,J=8.4Hz,1H,-Ph),8.85(d,J=8.4Hz,1H,-Ph),8.57(d,J=7.8Hz,1H,-Ph),8.38(d,J=8.4Hz ,1H,-Ph),7.76-7.78(m,1H,-Ph),7.62-7.72(m,5H,-Ph),6.90(d,J=8.4Hz,2H,-Ph),4.71(t,J=7.2Hz,2H,-CH2 -),4.33(s,2H,-CH2-),3.48-3.50(m,2H,-CH2-),3.20-3.24(m,4H,-CH2-),3.14(s,6H,N-CH3),3.03(s,6H,N-C H3),1.79(d,J=6.0Hz,2H,-CH2-),1.62(s,2H,-CH2-),1.06(t,J=7.2Hz,3H,-CH3),0.94(t,J=7.2Hz,3H,-CH3); 13 C NMR(150MHzDMSO-d6)δ:162.3,153.1,150.6,137.1,130.5,127.9,127.4,127.2,127.0,126.7,125.3,125.2,124.6,12 4.4,123.4,122.7,121.8,120.7,117.2,111.6,63.2,59.6,51.2,45.8,43.9,40.8,26.7,19.1,13.6,12.5; HRMS(ESI)C 35 H 44 BrN5O[M-Br] + calcd=550.3540; found=550.3547.

[0138] Example 23 Compound III18

[0139] Compound III18 was synthesized using the method described in Example 5. The physicochemical properties of compound III18 are as follows:

[0140] 1) White solid;

[0141] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (400MHz) characteristics:

[0142] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 59.6%. 1H NMR(600MHz DMSO-d6)δ:9.03(d,J=8.4Hz,1H,-Ph),8.92(d,J=7.8Hz,1H,-Ph),8.63(d,J=7.8Hz,-Ph),8.44(d,J=7.8Hz,1H,-Ph),7.82-7.84(m,1H,-Ph),7.6 9-7.80(m,5H,-Ph),6.95(d,J=8.4Hz,2H,-Ph),4.77(t,J=7.2Hz,2H,-CH2-),4.32(s,2H,-CH2-),3.49-3.52(m,2H,-CH2-),3.16(d,J=7.2Hz,2H,- CH2-),3.14(s,6H,N-CH3),3.10(d,J=8.4Hz,2H,-CH2-),3.06(s,6H,N-CH3),1.84-1.87(m,2H,-CH2-),1.60-1.65(m,2H,-CH2-),1.42-1.47(m,2 H,-CH2-),1.32-1.37(m,2H,-CH2-),1.23-1.27(m,2H,-CH2-),1.15-1.19(m,2H,-CH2-),0.88(t,J=7.2Hz,3H,-CH3),0.81(t,J=7.8Hz,3H,-CH3); 13 C NMR(150MHzDMSO-d6)δ:162.6,151.8,151.3,130.9,128.3,127.8,127.7,127.5,126.3,125.6,124.9,124.6,1 23.7,121.9,121.1,111.6,63.0,59.5,51.3,46.3,45.0,39.9,29.9,28.8,19.39,19.3,19.0,13.5; HRMS(ESI)C 39 H 52 BrN5O[M-Br] + calcd=606.4166; found=606.4174.

[0143] Example 24 Compound III19

[0144] Compound III19 was synthesized using the method described in Example 5. The physicochemical properties of compound III19 are as follows:

[0145] 1) Yellow solid;

[0146] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (400MHz) characteristics:

[0147] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 84.1%. 1 H NMR(600MHz DMSO-d6)δ:8.97(d,J=7.8Hz,1H,-Ph),8.85(d,J=8.4Hz,1H,-Ph),8.57(d,J=7.8Hz,1H,-Ph),8.39(d,J=8 .4Hz,1H,-Ph),7.76-7.79(m,1H,-Ph),7.60-7.72(m,5H,-Ph),6.89(d,J=9.0Hz,1H,-Ph),4.73(t,J=7.2Hz ,2H,-CH2-),4.29(s,2H,-CH2-),3.59-3.62(m,2H,-CH2-),3.12(s,6H,N-CH3),3.03(s,6H,N-CH3),2.16- 2.23(m,2H,-CH2-),1.22(d,J=3.6Hz,4H,-CH2-),1.09(d,J=6.6Hz,6H,-CH3),0.98(d,J=6.6Hz,6H,-CH3); 13 C NMR(150MHz DMSO-d6)δ:161.7,152.8,150.7,137.1,130.5,128.0,127.5,127.3,127.1,126.6,125.3,125.0,124.7,124.4,123.4,1 22.6,121.8,120.6,116.8,111.6,60.3,60.0,51.5,47.5,45.2,43.3,28.9,23.5,21.9,19.86,19.8,113.8; HRMS(ESI)C 39 H 52 BrN5O[M-Br] + calcd=578.3853; found=578.3860.

[0148] Example 25 Compound III20

[0149] Compound III20 was synthesized using the method described in Example 5. The physicochemical properties of compound III20 are as follows:

[0150] 1) White solid;

[0151] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (400MHz) characteristics:

[0152] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 80.7%. 1 H NMR(600MHz DMSO-d6)δ:8.98(d,J=7.8Hz,1H,-Ph),8.87(d,J=8.4Hz,1H,-Ph),8.59(d,J=7.8Hz,1H,-Ph),8.38(d,J=8.4Hz,1H, -Ph),7.77-7.79(m,1H,-Ph),7.64-7.74(m,5H,-Ph),6.91(d,J=8.4Hz,2H,-Ph),4.72(t,J=7.2Hz,2H,-CH2-),4.34 (s,2H,-CH2-),3.48-3.51(m,2H,-CH2-),3.15(s,6H,N-CH3),3.07(d,J=7.2Hz,2H,-CH2-),3.04(s,8H,-CH2-,N-CH 3),1.79-1.88(m,4H,-CH2-),1.61(d,J=3.6Hz,2H,-CH2-),0.83(d,J=6.6Hz,6H,-CH3),0.73(d,J=6.6Hz,6H,-CH3); 13 C NMR(150MHz DMSO-d6)δ:163.5,152.8,150.8,130.6,128.0,127.5,127.3,127.2,125.5,125.1,124.8,124.5,123.5, 122.6,121.8,120.8,111.6,62.9,59.5,53.6,52.3,51.4,45.9,26.8,25.7,19.6,19.3,19.2; HRMS(ESI)C 39 H 52 BrN5O[M-Br] + calcd=606.4166; found=606.4178.

[0153] Example 26 Compound III21

[0154] Compound III21 was synthesized using the method described in Example 5. The physicochemical properties of compound III21 are as follows:

[0155] 1) Brown solid;

[0156] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (400MHz) characteristics:

[0157] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 81.3%. 1H NMR(600MHz DMSO-d6)δ:8.96(d,J=8.4Hz,1H,-Ph),8.85(d,J=8.4Hz,1H,-Ph),8.65(t,J=4.8Hz,1H,-NH-),8.57(d,J=7.2Hz,1H,-Ph ),8.41(d,J=8.4Hz,1H,-Ph),7.76(t,J=7.2Hz,1H,-Ph),7.61-7.72(m,5H,-Ph),6.90(d,J=8.4Hz,2H,-Ph),4.73(t,J=7. 2Hz,2H,-CH2-),3.91(s,2H,-CH2-),3.25-3.28(m,2H,-CH2-),3.09(d,J=6.6Hz,2H,-Ph),3.04(s,6H,N-CH3),3.02(s,6 H,N-CH3),1.82-1.85(m,2H,-CH2-),1.48-1.53(m,2H,-CH2-),0.99(t,J=7.2Hz,4H,-CH2-),0.95(t,J=7.2Hz,3H,-CH3); 13 C NMR(150MHz DMSO-d6)δ:162.5,153.5,150.5,137.1,130.5,127.8,127.4,127.2,127.0,126.7,125.3,125.2,124.6,124.4,1 23.4,122.9,121.8,120.9,117.4,111.6,63.9,61.7,50.9,46.1,45.2,33.4,29.2,22.3,21.3,14.0; HRMS(ESI)C 34 H 42 BrN5O[M-Br] + calcd=536.3384; found=536.3388.

[0158] Example 27 Compound III22

[0159] Compound III22 was synthesized using the method described in Example 5. The physicochemical properties of compound III22 are as follows:

[0160] 1) Brown solid;

[0161] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (400MHz) characteristics:

[0162] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 95.1%. 1H NMR(600MHz DMSO-d6)δ:8.96(d,J=8.4Hz,1H,-Ph),8.85(d,J=8.4Hz,1H,-Ph),8.59(s,1H,-Ph),8.56-8.57(m,1H,-NH-),8.41(d,J=8 .4Hz,1H,-Ph),7.77-7.79(m,1H,-Ph),7.61-7.72(m,5H,-Ph),6.90(d,J=8.4Hz,2H,-Ph),4.73(t,J=7.2Hz,2H,-CH2-),3. 93(s,2H,-CH2-),3.25-3.28(m,2H,-CH2-),3.04(s,6H,N-CH3),3.02(s,6H,N-CH3),2.98-3.01(m,2H,-CH2-),1.82-1.84 (m,2H,-CH2-),1.48-1.53(m,4H,-CH2-),1.34-1.38(m,2H,-CH2-),1.01-1.04(m,2H,-CH2-),0.77(t,J=7.2Hz,3H,-CH3); 13 C NMR(150MHz DMSO-d6)δ:162.7,153.5,150.5,137.1,130.5,127.8,127.4,127.2,127.0,126.7,125.3,125.2,124.6,124.4,123 .4,122.9,121.8,120.9,117.4,111.6,63.9,61.7,50.9,46.1,40.2,39.9,29.2,22.3,21.7,21.4,11.1; HRMS(ESI)C 35 H 44 BrN5O[M-Br] + calcd=550.3540; found=550.3541.

[0163] Example 28 Compound III23

[0164] Compound III23 was synthesized using the method described in Example 5. The physicochemical properties of compound III23 are as follows:

[0165] 1) Yellow solid;

[0166] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (400MHz) characteristics:

[0167] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 82.0%. 1H NMR(400MHz DMSO-d6)δ:8.96(d,J=8.4Hz,1H,-Ph),8.84(d,J=8.0Hz,1H,-Ph),8.59(s,1H,-Ph),8.57(s,1H,-NH-),8.41(d,J= 8.4Hz,1H,-Ph),7.75(t,J=7.6Hz,1H,-Ph),7.60-7.72(m,5H,-Ph),6.90(d,J=8.8Hz,2H,-Ph),4.73(t,J=6.4Hz,2 H,-CH2-),3.93(s,2H,-CH2-),3.25-3.29(m,2H,-CH2-),3.04(s,6H,N-CH3),3.02(s,6H,N-CH3),1.81(d,J=8.0Hz ,2H,-CH2-),1.51(s,2H,-CH2-),1.16-1.20(m,4H,-CH2-),1.00-1.04(m,2H,-CH2-),0.79(t,J=6.8Hz,3H,-CH3); 13 CNMR(100MHz DMSO-d6)δ:162.8,153.6,150.7,137.2,130.6,127.9,127.5,127.2,127.1,126.8,125.4,125.3,124.4,123.5,123 .0,121.8,121.0,117.4,111.7,63.9,61.7,50.9,46.3,44.2,29.2,28.4,28.1,22.4,21.6,21.4,13.8; HRMS(ESI)C 37 H 47 BrN5O[M-Br] + calcd=578.3853; found=578.3857.

[0168] Example 29 Compound III24

[0169] Compound III24 was synthesized using the method described in Example 5. The physicochemical properties of compound III24 are as follows:

[0170] 1) Brown solid;

[0171] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (600MHz) characteristics:

[0172] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 53.4%. 1H NMR(600MHz DMSO-d6)δ:8.96(d,J=7.8Hz,1H,-Ph),8.85(d,J=8.4Hz,1H,-Ph),8.58-8.59(m,1H,-Ph),8.55(t,J=5.4Hz,,1H,-NH-),8.4 1(d,J=8.4Hz,1H,-Ph),7.77-7.79(m,1H,-Ph),7.61-7.72(m,5H,-Ph),6.90(d,J=9.0Hz,2H,-Ph),4.73(t,J=7.2Hz,2H,-CH2 -),3.92(s,2H,-CH2-),3.26-3.28(m,2H,-CH2-),3.04(s,6H,N-CH3),3.02(s,6H,N-CH3),1.81-1.85(m,2H,-CH2-),1.48-1. 53(m,2H,-CH2-),1.32-1.36(m,2H,-CH2-),1.13-1.28(m,8H,-CH2-),1.00-1.05(m,2H,-CH2-),0.80(t,J=7.2Hz,3H,-CH3); 13 C NMR(150MHz DMSO-d6)δ:163.2,154.0,151.1,137.7,131.1,128.4,128.0,127.7,127.6,127.3,125.9,125.8,125.2,124.9,123.9,123,4 ,122.3,121.4,117.9,112.2,64.4,62.3,51.4,46.6,40.4,38.9,31.2,29.8,28.9,26.3,22.8,22.4,21.9,14.3; HRMS(ESI)C 38 H 50 BrN5O[M-Br] + calcd=592.4010; found=592.4022.

[0173] Example 30 Compound III25

[0174] Compound III25 was synthesized using the method described in Example 5. The physicochemical properties of compound III25 are as follows:

[0175] 1) Brown solid;

[0176] 2) The nuclear magnetic resonance spectrum of this compound ( 1 HNMR (600MHz) characteristics:

[0177] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 90.8%. 1 H NMR(600MHz DMSO-d6)δ:8.96(d,J=8.4Hz,1H,-Ph),8.85(d,J=8.4Hz,1H,-Ph),8.57(d,J=7.8Hz,1H,-Ph),8.41(d,J=7.8Hz,1H,-Ph) ),7.76-7.79(m,1H,-Ph),7.61-7.72(m,5H,-Ph),6.90(d,J=8.4Hz,2H,-Ph),4.74(t,J=6.6Hz,2H,-CH2-),4.35(s,2H, -CH2-),3.67(s,2H,-CH2-),3.56(s,2H,-CH2-),3.07(s,8H,N-CH3,-CH2-),3.03(s,6H,N-CH3),2.75-2.78(m,2H,-CH2 -),2.62(s,2H,-CH2-),2.53(s,2H,-CH2-),1.82(t,J=7.2Hz,3H,-CH2),1.47-1.50(m,2H,-CH2-),0.99(m,2H,-CH2-); 13 C NMR(150MHz DMSO-d6)δ:162.0,153.5,150.5,137.1,130.5,127.8,127.4,127.2,127.0,126.7,125.3,125.2,124.6,124.4,123 .4,122.9,121.7,120.9,117.4,111.6,64.1,60.0,50.9,47.1,46.2,43.7,29.2,26.5,26.2,22.4,21.4; HRMS(ESI)C 36 H 44 BrN5OS[M-Br] + calcd=594.3266; found=594.3266.

[0178] Example 31 Compound III26

[0179] Compound III26 was synthesized using the method described in Example 5. The physicochemical properties of compound III26 are as follows:

[0180] 1) Brown solid;

[0181] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (600MHz) characteristics:

[0182] Using CDCl3 as solvent, the peaks were assigned as follows: Yield: 51.5%, brown solid. 1 H NMR(600MHzDMSO-d6)δ:8.96(d,J=8.4Hz,1H,-Ph),8.85(d,J=8.4Hz,1H,-Ph),8.58(d,J=7.8Hz,1H,-Ph),8.41(d,J =7.8Hz,1H,-Ph),7.76(t,J=7.2Hz,1H,-Ph),7.61-7.72(m,5H,-Ph),6.90(d,J=8.4Hz,2H,-Ph),4.73(t,J=6.6Hz,2H ,-CH2-),4.25(s,2H,-CH2-),3.17-3.24(m,6H,-CH2-),3.07(s,6H,N-CH3),3.03(s,6H,N-CH3),1.80-1.85(m,2H,- CH2-),1.45-1.50(m,2H,-CH2-),1.04(t,J=7.2Hz,3H,-CH3),0.99-1.03(m,2H,-CH2-),0.94(t,J=6.6Hz,3H,-CH3); 13 C NMR(150MHz DMSO-d6)δ:162.9,154.1,151.1,137.7,131.1,128.4,128.0,127.7,127.6,127.3,125.9,125.8,125.2,124.9,124 .0,123.4,122.3,121.4,117.9,112.2,64.3,60.2,51.5,46.7,41.4,40.4,29.8,22.9,22.0,14.2,13.0; HRMS(ESI)C 36 H 46 BrN5O[M-Br] + calcd=564.3697; found=564.3703.

[0183] Example 32 Compound III27

[0184] Compound III27 was synthesized using the method described in Example 5. The physicochemical properties of compound III27 are as follows:

[0185] 1) Yellow solid;

[0186] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (600MHz) characteristics:

[0187] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 73.4%. 1 H NMR(600MHz DMSO-d6)δ:8.96(d,J=8.4Hz,1H,-Ph),8.85(d,J=8.4Hz,1H,-Ph),8.58-8.59(m,1H,-Ph),8.40(d,J=8.4Hz,1H,-Ph),7.76-7. 78(m,1H,-Ph),7.61-7.72(m,5H,-Ph),6.90(d,J=9.0Hz,2H,-Ph),4.73(t,J=7.2Hz,2H,-CH2-),4.28(s,2H,-CH2-),3.40(s,2H ,-CH2-),3.13(t,J=7.2Hz,2H,-CH2-),3.07-3.10(m,8H,N-CH3,-CH2-),3.03(s,6H,N-CH3),1.79-1.84(m,2H,-CH2-),1.43-1. 47(m,4H,-CH2-),1.37-1.40(m,2H,-CH2-),0.99-1.04(m,2H,-CH2-),0.78(t,J=7.2Hz,3H,-CH3),0.71(t,J=7.2Hz,3H,-CH3); 13 C NMR(150MHzDMSO-d6)δ:163.4,154.0,151.1,131.1,128.4,128.0,127.7,127.7,127.6,127.3,125.9,125.8,125.2,124.9,124.0,12 3.4,122.3,121.4,117.8,112.2,63.9,60.0,50.1,51.7,48.5,47.4,46.8,44.0,29.8,22.9,22.0,21.6,20.6,11.4,11.3,HRMS(ESI)C 38 H 50 BrN5O[M-Br] + calcd=592.4010; found=592.4017.

[0188] Example 33 Compound III28

[0189] Compound III28 was synthesized using the method described in Example 5. The physicochemical properties of compound III28 are as follows:

[0190] 1) Yellow solid;

[0191] 2) The nuclear magnetic resonance spectrum of this compound ( 1H NMR (600MHz) characteristics:

[0192] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 90.8%. 1 H NMR(600MHz DMSO-d6)δ:8.97(d,J=8.4Hz,1H,-Ph),8.86(d,J=8.4Hz,1H,-Ph),8.58(d,J=7.8Hz,1H,-Ph),8.41(d,J=7.8Hz,1H,-Ph),7.76-7.78(m,1H,-Ph), 7.62-7.73(m,5H,-Ph),6.90(d,J=8.4Hz,2H,-Ph),4.74(t,J=7.2Hz,2H, -CH2-),4.27(s,2H,-CH2-),3.38-3.40(m,2H,-CH2-),3.16-3.19(m,2H,- CH2-),3.10(t,J=7.8Hz,2H,-CH2-),3.08(s,6H,N-CH3),3.03(s,6H,N-CH3),1.80-1.84(m,2H,-CH2-),1.45-1.48(m,2H,-CH2-),1.33-1.37(m,2 H,-CH2-),1.21-1.23(m,2H,-CH2-),1.13-1.17(m,2H,-CH2-),1.0o-1.04(m,2H,-CH2-),0.84(t,J=7.2Hz,3H,-CH3),0.78(t,J=7.2Hz,3H,-CH3); 13 C NMR(150MHz DMSO-d6)δ:163.6,162.7,150.6,130.6,127.9,127.4,127.2,127.1,125.4,125.3,124.7,124.4,123.4,122.8,121.8,1 20.9,111.6,63.4,59.5,56.6,51.1,46.2,45.0,44.9,43.4,29.9,29.2,28.8,22.4,21.5,19.3,19.2,13.5,HRMS(ESI)C 40 H 54 BrN5O[M-Br] + calcd=620.4323; found=620.4326.

[0193] Example 34 Compound III29

[0194] Compound III29 was synthesized using the method described in Example 5. The physicochemical properties of compound III29 are as follows:

[0195] 1) Yellow solid;

[0196] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (600MHz) characteristics:

[0197] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 56.2%. 1 H NMR(600MHz DMSO-d6)δ:8.97(d,J=8.4Hz,1H,-Ph),8.85(d,J=8.4Hz,1H,-Ph),8.57(d,J=7.8Hz,1H,-Ph),8.37(d,J=8.4Hz, 1H,-Ph),7.75-7.77(m,1H,-Ph),7.62-7.72(m,5H,-Ph),6.90(d,J=8.4Hz,2H,-Ph),4.72(t,J=7.2Hz,2H,-CH2-) ,4.29(s,2H,-CH2-),3.48-3.51(m,2H,-CH2-),3.42-3.44(m,2H,-CH2-),3.12(s,6H,N-CH3),3.03(s,6H,N-CH3 ),1.78(t,J=6.6Hz,2H,-CH2-),1.59(s,2H,-CH2-),1.16-1.19(m,8H,-CH3,-CH2-),1.09(d,J=6.6Hz,6H,-CH3); 13 C NMR(150MHzDMSO-d6)δ:162.4,153,6,151.2,131.1,128.5,128.0,127.7,127.6,127.2,125.9,125.8,125.2,125.0,124 .0,123.3,122.3,121.3,112.2,63.5,60,9,58.3,52.0,48.2,46.4,45.8,44.1,27.3,20.6,20.5,20.4,19.7; HRMS(ESI)C 38 H 50 BrN5O[M-Br] + calcd=592.4010; found=592.4013.

[0198] Example 35 Compound 30

[0199] Compound III30 was synthesized using the method described in Example 5. The physicochemical properties of compound III30 are as follows:

[0200] 1) Yellow solid;

[0201] 2) The nuclear magnetic resonance spectrum of this compound ( 1 H NMR (600MHz) characteristics:

[0202] Using CDCl3 as solvent, the peak assignments are as follows: Yield: 90.8%. 1 H NMR(600MHz DMSO-d6)δ:8.98(d,J=7.8Hz,1H,-Ph),8.87(d,J=8.4Hz,1H,-Ph),8.59(d,J=7.8Hz,1H,-Ph),8.42(d,J=8.4Hz,1H,-Ph),7. 77-7.79(m,1H,-Ph),7.62-7.74(m,5H,-Ph),6.91(d,J=9.0Hz,2H,-Ph),4.75(t,J=6.6Hz,2H,-CH2-),4.28(s,2H,-CH2-),3. 38-3.40(m,2H,-CH2-),3.09(s,6H,N-CH3),3.05(s,2H,-CH2-),3.04(s,6H,N-CH3),2.99(d,J=7.8Hz,2H,-CH2-),1.79-1.85 (m,4H,-CH2-),1.41-1.47(m,2H,-CH2-),0.98-1.06(m,2H,-CH2-),0.79(d,J=6.6Hz,6H,-CH3),0.66(d,J=6.6Hz,6H,-CH3); 13 C NMR(150MHzDMSO-d6)δ:164.5,163.6,150.7,130.6,128.0,127.5,127.3,127.2,125.6,125.2,124.9,124.4,123.5,122.6,12 1.8,121.0,111.6,63.1,59.4,53.5,52.2,51.4,46.3,43.3,29.1,26.7,26.2,25.6,22.4,21.6,19.7,19.5,19.3; HRMS(ESI)C 40 H 54 BrN5O[M-Br] + calcd=620.4323; found=620.4330.

[0203] Application Example 1: In vitro antibacterial activity assay

[0204] 1. Test bacteria:

[0205] Staphylococcus aureus (ATCC 29213); methicillin-resistant Staphylococcus aureus (MRSA).

[0206] 2. Samples and reagents:

[0207] The samples were: phenanthrenequinone, vancomycin, meropenem, and compounds III1-30 prepared in the examples.

[0208] 3. Testing Method:

[0209] According to the Clinical Laboratory Standards Institute (CLSI) standards, the in vitro antibacterial activity of phenanthrenequinone, compounds 1-32 of this invention, and clinical antibacterial drugs vancomycin and meropenem was tested using a 96-well plate and serial dilution method. The minimum drug concentration observed with the naked eye in the smallest completely clear well was defined as the MIC value.

[0210] Table 1. In vitro antibacterial activity (μg / mL) of the phenanthrimidazole-amine copolymer III1-30 of the present invention

[0211]

[0212]

[0213]

[0214] Note: a Ec: Escherichia coli ATCC25922; b Sa: Staphylococcus aureus (ATCC29213); c M-1-10: 10 clinically isolated MRSA strains; d SI: Selectivity Index (HC) 50 / MICs of S.aureus); e Van: Vancomycin; f MEM: Meropenem; g ND: Not determined; experiment repeated 3 times.

[0215] First, the antibacterial activity of phenanthrimidazole-amine copolymers III1-30 against Gram-positive (G+) bacteria *Staphylococcus aureus* ATCC 29213, Gram-negative (G-) bacteria *Escherichia coli* ATCC 25922, and ten clinical MRSA isolates was evaluated using the broth microdilution method. As shown in Table 1, most phenanthrimidazole-amine copolymers exhibited stronger antibacterial activity against all tested strains, especially Gram-positive (G+) bacteria, than their precursor phenanthrenequinone. Among these derivatives, III13, III18, and III30 showed antibacterial efficacy comparable to the positive control drug vancomycin, with MIC values ​​of 1 μg / mL against ATCC 29213 and 0.5–2 μg / mL against MRSA. Among them, compound III13 exhibited the strongest anti-MRSA activity, with a MIC of 0.5 μg / mL against five clinical MRSA isolates, superior to vancomycin (MIC = 1 μg / mL). Simultaneously, compound III13 showed good bactericidal activity against *Escherichia coli* ATCC 25922, with a MIC of 32 μg / mL. HC 50 The value represents the concentration required for a compound to lyse 50% of red blood cells (RBCs) within 1 hour and is commonly used to assess antimicrobial toxicity against eukaryotic cells. As shown in Table 1, most target compounds exhibited superior membrane selectivity compared to the parent phenanthrenequinone. Among them, III13 showed lower hemolytic toxicity (HC). 50 With a concentration of 895.1 μg / mL and optimal membrane selectivity (SI = 859.1), which are 2.68-fold and 10.7-fold higher than the parent phenanthrenequinone, respectively, it is expected to become a clinical antibacterial drug against methicillin-resistant Staphylococcus aureus.

[0216] Application Example 2: Time-based sterilization kinetics experiment:

[0217] 1. Test bacteria:

[0218] Staphylococcus aureus ATCC 29213; MRSA-4 (clinical isolate).

[0219] 2. Samples and reagents:

[0220] The samples were: vancomycin and compound III13 prepared in the examples.

[0221] 3. Testing Method:

[0222] The dynamic bactericidal curves of compound III13 against Staphylococcus aureus ATCC 29213 and MRSA-4 were determined using the plate count method. First, the bacteria were incubated in MHB at 37°C and 180 rpm for 3–5 hours, then diluted to 1.0 × 10⁻⁶. 5 CFU / mL. Different concentrations of compound III13 (8×MIC, 4×MIC) and vancomycin (8×MIC) were added to the bacterial suspension, and 100 μL of the suspension was serially diluted at 0, 0.5, 1, 2, 4, 6, and 8 hours. The suspensions were then placed on MHB agar plates and incubated at 37°C for 16–18 hours. Colony counts were performed, and a logarithmic curve of colony count versus time was plotted. Untreated bacterial suspensions served as a blank control, and each group was repeated three times. Figure 1 As shown in A and B, compound III13 exhibits time- and dose-dependent bactericidal properties and rapid bactericidal activity. Specifically, at a concentration of 4×MIC, compound III13 completely killed *Staphylococcus aureus* ATCC 29213 within 2 hours; at a concentration of 8×MIC, it completely killed *Staphylococcus aureus* ATCC 29213 within 1 hour. Excitingly, compound III13 also showed highly effective bactericidal properties against clinical MRSA-4 isolates, completely killing the bacteria within 4 hours (8×MIC) and 6 hours (4×MIC), respectively. In contrast, the positive control vancomycin at a concentration of 8×MIC only inhibited the growth of both *Staphylococcus aureus* ATCC29213 and MRSA-4, failing to completely kill the bacteria. These results indicate that compound III13 can rapidly kill bacteria in a time- and dose-dependent manner, shortening the treatment time for bacterial infections, and possesses greater potential and rapid bactericidal activity compared to the antibiotic vancomycin.

[0223] Application Example 3: Fluorescence Properties Study

[0224] The spectra of the compound in PBS, 0.9% NaCl, and DMSO were characterized using a UV-Vis spectrophotometer and a fluorescence spectrophotometer to determine the absorption wavelength range of compound III13. Single colonies of MRSA-4 were incubated in 2.0 mL LB broth for 5–6 hours (37 °C), centrifuged at 4 °C for 5 minutes, and washed twice with PBS. The bacterial solution was resuspended in PBS containing 32 μg / mL III13 and incubated for 3 hours (37 °C, 180 rpm). Bacterial solutions were collected in the dark every 20 minutes. The fluorescence absorption curves of the compound were measured using a microplate reader, with excitation wavelength at 260 nm and emission wavelengths at 300–700 nm. Each group was repeated three times, with untreated bacterial solutions serving as blank controls. The physical spectroscopic characterization results showed that ( Figure 2The maximum UV absorption wavelength of compound III13 is 257 nm, and its fluorescence intensity in PBS is higher than that in 0.9% NaCl and DMSO. Figure 2 A, B). Further spectral monitoring of the dynamic interaction between the test compounds and bacteria revealed that the fluorescence signal of compound III13 (MIC = 0.5-1 μg / mL) significantly increased at 20 minutes, and the fluorescence intensity at 180 minutes was approximately 1.9 times higher than at 0 minutes. Figure 2 C). However, the fluorescence signal of compound II2 (MIC > 64 μg / mL), which had lower antibacterial activity, showed almost no change within 180 minutes. Figure 2 D). This phenomenon indicates that the introduction of tertiary amine fragments can significantly enhance the interaction between the compound and bacteria, thereby enhancing the fluorescence signal and potentially dynamically presenting the bactericidal process, thus achieving integrated diagnosis and treatment.

[0225] Application Example 4: In vivo safety evaluation experiment

[0226] 1. Reagents:

[0227] Compound III13 prepared in the example, 0.9% NaCl.

[0228] 2. Test animals

[0229] SPF-grade KM mice (purchased from Beijing Spaford Biotechnology Co., Ltd., weighing 19-22g, 4-6 weeks old).

[0230] 3. Testing Methods

[0231] Animal experiments were conducted according to the "Guidelines for the Care and Use of Laboratory Animals" approved by the Animal Protection and Utilization Committee. Thirty mice were divided into groups of five: Control group (0.9% NaCl), III13 (5 mg / kg), III13 (10 mg / kg), III13 (20 mg / kg), III13 (40 mg / kg), and III13 (80 mg / kg). Hair was removed from the backs of the mice using a depilatory cream. After 24 hours of feeding, the mice were anesthetized with a small amount of ether, followed by an injection of 60 μL of 0.9% NaCl and different doses of compound III13. After 24 hours, the mice were observed for mortality and abnormal skin phenomena such as ulceration and swelling. Blood was collected from the eyes of mice in the highest dose group (without adverse reactions) for routine blood tests and blood biochemical parameters, including platelet count (PLT), hemoglobin (HGB), red blood cell count (RBC), mean corpuscular volume (MCV), hematocrit (HCT), and white blood cell count (WBC). Serum was collected for blood biochemistry tests, including urea (UREA), albumin (ALB), and creatinine (CREA), to evaluate the in vivo safety of compound III13 in mice.

[0232] Compared to the control group, mice in the III13 groups (80 and 40 mg / kg) showed slight redness and swelling at the administration site, while no abnormalities were observed in the III13 groups (20, 10, and 5 mg / kg). Furthermore, as... Figure 3 As shown, compared with the blank control group, compound III13 (20 mg / kg) showed no significant differences in blood biochemical and complete blood count indicators. These results indicate that compound III13 has good biocompatibility in mice, and a safe dose of 20 mg / kg can be selected for further in vivo anti-MRSA evaluation.

[0233] Application Example 5: In vivo anti-MRSA infection activity experiment

[0234] Establish a mouse skin abscess model infected with MRSA-4

[0235] Thirty SPF-grade KM mice were acclimatized for one week. The mice were then randomly divided into four groups of six: a blank control group, a model group, a positive control vancomycin group, a high-dose treatment group, and a low-dose treatment group. The clinical isolate MRSA-4 in its logarithmic growth phase was resuspended in sterile PBS at a concentration of 6 × 10⁻⁶ mg / L. 8 CFU / mL was injected subcutaneously into the back of each mouse at a volume of 60 μL, while the control group was injected with the same volume of 0.9% NaCl.

[0236] Anti-MRSA infection activity study

[0237] Two hours later, mice in each group were injected with 60 μL of different doses of compound III13 (5 mg / kg and 10 mg / kg), the positive control drug vancomycin (5 mg / kg), and physiological saline, respectively, into the abscess site. The survival status of the mice was observed after 24 hours. The mice were then euthanized by dislocation, and the infected skin was dissected in a biosafety cabinet and added to 1 mL of sterile physiological saline. The mixture was then ground to a fragmented state using a tissue homogenizer (4℃, 70 Hz). The tissue fluid from each group was then serially diluted in 96-well plates. 10 μL of each diluted sample was aspirated and dropped onto LB agar plates. Three parallel controls were maintained for each group. The plates were air-dried and incubated at 37℃ for 18-24 hours, after which the colony count was recorded. Figure 4 As shown, compared with the model group, vancomycin (5 mg / kg) and III13 (5 mg / kg and 10 mg / kg) significantly reduced the number of MRSA-4 bacteria in mouse skin. Specifically, the bacterial survival rate at the skin infection site in mice treated with III13 (10 mg / kg) was only 0.01%. However, at the same dose (5 mg / kg), compound III13 was more effective than vancomycin, with MRSA-4 survival rates of 0.42% and 2.15%, respectively. Compound III13 demonstrated high biocompatibility and strong in vivo anti-infective activity in a mouse model of MRSA-infected skin abscesses, and is expected to be developed into a novel antibacterial agent for the treatment of MRSA infection.

Claims

1. A fluorescent phenanthrimidazole-amine copolymer or a pharmaceutically acceptable salt thereof, said copolymer having the structural formula shown in formula (III): ； in, n = 3, 4, or 5, R is selected from or In this context, R1 and R2 are independently selected from hydrogen or C1-C6 alkyl groups; A is selected from C, N, S or O.

2. The phenanthrimidazole-amine copolymer or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, Where n = 3, 4, or 5, and R is selected from... 、 。 3. The phenanthrimidazole-amine copolymer or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, The copolymer is selected from the following compounds: 。 4. The phenanthrimidazole-amine copolymer or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, The fluorescent phenanthreneimidazole-amine copolymer is selected from the following compounds: ； Wherein, n and R are as described in any one of claims 1-3.

5. The method for preparing the phenanthrimidazole-amine copolymer according to any one of claims 1-4, characterized in that, Includes the following steps: (1) Under alkaline conditions, bromoacetyl bromide is reacted with RH to obtain intermediate 1; (2) Under alkaline conditions, intermediate 1 continues to react with dimethylamine to generate small molecule peptide fragment 2; (3) Using phenanthrenequinone as a substrate, it was reacted with p-diethylaminobenzaldehyde and ammonium acetate in a three-component Debus-Radziszewski imidazole synthesis reaction to obtain fluorescent N,N-dimethyl-4-(1H-phenanthrene[9,10-d]imidazol-2-yl)aniline, which is intermediate I; (4) Under alkaline conditions, intermediate I is reacted with dibromoalkane to synthesize intermediate II; (5) Finally, intermediate II is reacted with intermediate 2 under alkaline conditions to generate a series of phenanthrimidazole-amine copolymers; ； Wherein, n and R are as described in any one of claims 1-4.

6. The method for preparing phenanthrimidazole-amine copolymers according to claim 5, characterized in that, In step (1), the base in the alkaline conditions is K2CO3, the reaction solvent is selected from dichloromethane solution, the reaction temperature is 0±2 ℃, and the reaction time is 5h−12 h; In step (2), the base in the alkaline conditions is K2CO3, the reaction solvent is selected from acetone solution, the reaction temperature is 95-100℃, and the reaction time is 1-2 h.

7. The method for preparing phenanthrimidazole-amine copolymers according to claim 5, characterized in that, In step (3), the reaction temperature is 100±2 ℃, the reaction time is 1 h−2 h, and the reaction solvent is glacial acetic acid solution; In step (4), the base in the alkaline conditions is K2CO3, the molar ratio of intermediate I to the base is 1:4−1:5, the molar ratio of intermediate I to dibromoalkane is 1:4−1:8, the reaction temperature is 45−60 ℃, the reaction time is 12 h−14 h, and the reaction solvent is anhydrous acetonitrile. In step (5), the base in the alkaline conditions is K2CO3, the molar ratio of intermediate II to the base is 1:4-1:5, the molar ratio of intermediate II to intermediate 2 is 1:4-1:5, the reaction temperature is 70-80 ℃, the reaction time is 12 h-16 h, and the reaction solvent is ethanol.

8. The use of the phenanthrimidazole-amine copolymer or a pharmaceutically acceptable salt thereof according to any one of claims 1-4 in the preparation of antibacterial drugs, characterized in that, The antibacterial drug is used to inhibit Staphylococcus aureus or methicillin-resistant Staphylococcus aureus.

9. The use of the phenanthrimidazole-amine copolymer or a pharmaceutically acceptable salt thereof according to any one of claims 1-4 in the preparation of a therapeutic antibacterial agent, characterized in that, The antibacterial agent is used to inhibit Staphylococcus aureus or methicillin-resistant Staphylococcus aureus.

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