Pyridine quaternary ammonium salt-containing rutaecarpin derivative as well as preparation method and antibacterial application thereof

Abstract

The invention discloses a series of rutaecarpin derivatives containing pyridine quaternary ammonium salt as well as a preparation method and antibacterial application of the rutaecarpin derivatives. The series of compounds take a natural product rutaecarpin as a parent structure, a pyridine ring is introduced and then connected with a chloroacetamide fragment, so that a series of rutaecarpin derivatives containing pyridine quaternary ammonium salt are prepared, and the structural general formula of the rutaecarpin derivatives containing pyridine quaternary ammonium salt is as shown in formula (1). The series of compounds have relatively strong in vivo and in vitro antibacterial activity on Staphylococcus aureus ATCC 29213 and clinically isolated methicillin-resistant Staphylococcus aureus (Methicillin-Resistance S. aureus, MRSA), and can be used for preparing the antibacterial agent for treating the staphylococcus aureus, the antibacterial agent for treating the staphylococcus aureus, the antibacterial agent for treating the methicillin-resistant Staphylococcus aureus and the antibacterial agent for treating the methicillin-resistant Staphylococcus aureus, the antibacterial agent for treating the staphylococcus aureus, the antibacterial agent for treating the staphylococcus aureus and the antibacterial agent for treating the staphylococcus aureus. Through further structural optimization, the water solubility and antibacterial activity of the compound are enhanced, and the compound has relatively low in-vivo and in-vitro toxicity, safe hemolytic activity and low drug resistance. Therefore, the rutaecarpin derivatives containing the pyridine quaternary ammonium salt have the potential of being further developed into novel antibacterial drugs. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical chemistry, and in particular relates to an evodia rutaecarpine derivative containing a quaternary pyridinium ammonium salt, a preparation method thereof and an antibacterial application thereof. Background Art

[0002] Bacterial pathogens are the main cause of human disease and death, and antibiotics are mainly used to treat infections caused by various bacteria. Due to the abuse of antibiotics, many pathogens have evolved resistance to major types of antibiotics. (MacLean R C, San Millan A. The evolution of antibiotic resistance [J]. Science, 2019, 365 (6458): 1082-1083.) Staphylococcus aureus (S. aureus) is a representative of Gram-positive bacteria. After infection, it can induce a variety of diseases, from skin and soft tissue infections to bloodstream and internal organ infections, sepsis, pneumonia, meningitis, etc. Currently, more than 60% of Staphylococcus aureus isolates are resistant to methicillin (methicillin-resistant Staphylococcus aureus / MRSA). MRSA has an in vivo gene encoding PBP2a, which has low sensitivity to β-lactam antibiotics and can synthesize cell walls at lethal β-lactam concentrations. Peptidoglycan is the core component of bacterial cell walls. PBP2a enables bacteria to maintain peptidoglycan biosynthesis and complete bacterial reproduction. (Ambade SS, Gupta VK, Bhole RP, et al. A review on five and six-membered heterocyclic compounds targeting the penicillin-binding protein2 (PBP2A) of Methicillin-resistant Staphylococcus aureus (MRSA) [J]. Molecules, 2023, 28 (20): 7008.) Antibiotic resistance has caused serious health and economic problems, and the development of new antibacterial drugs with potential therapeutic effects on MRSA infections is urgent.

[0003] Quaternary ammonium salts are also called quaternary ammonium salts. They are easily soluble in water and have certain biological activity. Quaternary ammonium salts are amphiphilic substances with good antibacterial properties. They can kill bacteria by electrostatic adsorption or insertion into cell membranes / changing cell membrane permeability, thereby reducing the probability of bacterial resistance. (Zhou Z, Zhou S, Zhang X, et al. Quaternary ammonium salts: insights into synthesis and new directions in antibacterial applications[J]. Bioconjugate Chemistry, 2023, 34(2): 302-325.) Rutaecarpine (Rut) is derived from the dried nearly mature fruit of the Rutaceae plant Evodia rutaecarpa (Juss.) Benth. There are many clinical studies on the treatment of gastrointestinal diseases, anti-tumor, hypertension, etc., but few studies on antibacterial properties. In addition, its water solubility and bioavailability are poor, making it difficult to directly become an antibacterial drug (Baburin I, Varkevisser R, Schramm A, et al. Dehydroevodiamine and hortiamine, alkaloids from the traditional Chinese herbal drug Evodia rutaecarpa, are IKr blockers with proarrhythmic effects in vitro and in vivo[J]. Pharmacological research, 2018, 131: 150-163.), therefore, it is necessary to modify its structure to improve its water solubility and antibacterial activity. Summary of the invention

[0004] Purpose of the invention: In view of the above technical problems, the present invention provides a quaternary ammonium salt-containing evodia rutaecarpine derivative, a preparation method thereof and an antibacterial application thereof. The present invention draws on the characteristics of quaternary ammonium salts as cationic surfactants with antibacterial effects, uses the natural product evodia rutaecarpine as a parent, modifies its structure, first introduces a pyridine ring, and then connects it with a chloroacetamide fragment to form a quaternary ammonium salt, aiming to obtain a highly water-soluble, highly active, and low-toxic evodia rutaecarpine derivative containing a quaternary ammonium salt of pyridine. The evodia rutaecarpine derivative containing a quaternary ammonium salt of pyridine has a good antibacterial effect on Staphylococcus aureus (S. aureus) ATCC 29213 and clinically isolated MRSA strains and MRSA standard strains N315, and solves the problems of high toxicity and poor water solubility.

[0005] Technical solution: In order to achieve the above-mentioned invention object, the technical solution adopted by the present invention is as follows:

[0006] The quaternary ammonium salt-containing evodia rutaecarpine derivative has a structure as shown in the following formula (1):

[0007]

[0008] Where n = 3, 4 or 5, R is Among them, R 1 and R 2 Independently selected from H, phenyl, 4-6 membered cycloalkyl or C1-C5 alkyl, ring A is a 4-6 membered heterocycloalkyl containing one or two heteroatoms.

[0009] As a preferred embodiment, the heteroatoms in the ring A, except the N atom connected to the parent nucleus, are absent or selected from one of N, O and S.

[0011] As a preferred solution, n, R is one of the following combinations:

[0012] (1) n = 3, R = (2) n = 3, R = (3) n = 3, R = (4) n = 3, R = (5) n = 3, R = (6) n = 3, R = (7) n = 3, R = (8) n = 3, R = (9) n = 3, R = (10) n = 3, R = (11) n = 4, R = (12) n = 4, R = (13) n = 4, R = (14) n = 4, R = (15) n = 4, R = (16) n = 4, R = (17) n = 4, R = (18) n = 4, R = (19) n = 4, R = (20)n=4,R= (21) n = 5, R = (22) n = 5, R = (23) n = 5, R = (24) n = 5, R = (25)n=5,R= (26)n=5,R= (27)n=5,R= (28)n=5,R= (29)n=5,R= (30)n=5,R=

[0013] The above specific selections represent compounds 1-30 in the following examples (each compound corresponds to the same serial number as above).

[0014] The present invention also provides a method for preparing the evodia rutaecarpine derivative containing a quaternary pyridinium ammonium salt, comprising the following steps:

[0015] (1) Using evodia rutaecarpine 1 as a substrate, it reacts with dibromoalkane 5 to obtain monobromoalkane-substituted intermediate 2;

[0016] (2) intermediate 2 reacts with 4-mercaptopyridine to generate intermediate 3;

[0017] (3) Chloroacetamide 6 reacts with intermediate 3 under the action of a catalyst to generate the quaternary pyridine ammonium salt-containing evodia rutaecarpine derivative 4, as shown in the following reaction formula:

[0018]

[0019] Wherein, n and R are the same as described above.

[0020] As a specific implementation scheme, in step (1), the reaction molar ratio of the evodia rutaecarpine 1 to the dibromoalkane 5 is 1:(7.0-12.0), the catalyst is potassium carbonate, the reaction molar ratio of the evodia rutaecarpine 1 to potassium carbonate is 1:(6.5-7.0), the reaction temperature is 45±55°C, and the reaction solvent is acetone;

[0021] As a specific implementation scheme, in step (2), the reaction molar ratio of the intermediate 2 to 4-mercaptopyridine is 1:(1.0-1.5), the catalysts are potassium carbonate and potassium iodide, the reaction molar ratio of the intermediate 2 to potassium carbonate is 1:(1.0-1.7), and the reaction molar ratio with potassium iodide is 1:(0.01-0.03), the reaction temperature is 45±50°C, and the reaction solvent is acetone.

[0022] As a specific implementation scheme, in step (3), the preparation method of chloroacetamide 6 comprises the following steps: chloroacetyl chloride and aliphatic amines / aromatic amines containing different substituents undergo a substitution reaction under alkaline catalyst conditions to generate chloroacetamide 6:

[0023]

[0024] Preferably, the reaction molar ratio of the chloroacetyl chloride to the aliphatic amine / aromatic amine containing different substituents is (1.5-1.7):1, the alkaline catalyst is triethylamine, the reaction molar ratio of the aliphatic amine / aromatic amine containing different substituents to triethylamine is (1.0-1.2):1.5, the reaction temperature is 0±5°C, and the reaction solvent is anhydrous dichloromethane.

[0025] More preferably, the reaction molar ratio of chloroacetyl chloride to aliphatic amines / aromatic amines containing different substituents is 1.5:1, the alkaline catalyst is triethylamine, the reaction molar ratio of aliphatic amines / aromatic amines containing different substituents to triethylamine is 1:1.5, the reaction temperature is 0°C, and the reaction solvent is anhydrous dichloromethane.

[0026] As a specific implementation scheme, in step (3), the reaction molar ratio of the intermediate 3 to chloroacetamide 6 is 1:(2.0-2.5), nitrogen protection is adopted, the reaction temperature is 75±80°C, and the reaction solvent is anhydrous acetonitrile.

[0027] Furthermore, as a preferred solution:

[0028] In step (1), the reaction molar ratio of evodia rutaecarpine to dibromoalkane is 1:12, the catalyst is potassium carbonate, the reaction molar ratio of evodia rutaecarpine to potassium carbonate is 1:7, the reaction temperature is 50° C., and the reaction solvent is acetone.

[0029] In step (2), the reaction molar ratio of the intermediate 2 to 4-mercaptopyridine is 1:1, the catalysts are potassium carbonate and potassium iodide, the reaction molar ratio of the intermediate 2 to potassium carbonate is 1:1.5, and the reaction molar ratio to potassium iodide is 1:0.02, the reaction temperature is 50°C, and the reaction solvent is acetone.

[0030] In step (3), the reaction molar ratio of the intermediate 3 to chloroacetamide is 1:2, nitrogen protection is adopted, the reaction temperature is 80° C., and the reaction solvent is anhydrous acetonitrile.

[0031] Finally, the present invention provides the use of the quaternary ammonium salt-containing evodia rutaecarpine derivative in the preparation of an antibacterial drug. Preferably, the antibacterial drug can inhibit Staphylococcus aureus or methicillin-resistant Staphylococcus aureus. For example, it can inhibit Staphylococcus aureus ATCC 29213 and various clinical methicillin-resistant Staphylococcus aureus MRSA.

[0032] The compounds of the present invention effectively connect the natural product evodipine with chloroacetamide fragments containing different substituents to generate quaternary ammonium salts. Drawing on the characteristics of quaternary ammonium salts as cationic surfactants with antibacterial effects, a series of evodipine derivatives containing pyridine quaternary ammonium salts are designed and synthesized. The hydrophobic part in the evodipine-quaternary ammonium salt compounds is conducive to the insertion of the compounds into the bacterial phospholipid bilayer membrane, and the positively charged cationic part is conducive to the interaction with the negatively charged bacterial cell membrane, thereby causing bacterial death. The present invention evaluates the in vitro antibacterial activity of all target compounds, and finds that all target compounds show good in vitro antibacterial activity against S. aureus ATCC 29213 and clinically isolated MRSA strains, with a minimum inhibitory concentration (MIC≤64μg / mL).

[0033] In particular, the preferred compound 14 has the most outstanding anti-MRSA activity in vitro and in vivo, and its antibacterial effect in vivo is comparable to that of vancomycin at the same dose. In addition, the target compound has safe hemolytic activity, low toxicity, and plasma stability. Therefore, this type of compound provides certain inspiration for the study of new anti-MRSA drugs and has broad application prospects.

[0034] Technical effect: The quaternary ammonium pyridinium salt-containing evodia rutaecarpine derivative prepared by the present invention has good in vitro and in vivo antibacterial effect on S. aureus ATCC29213 and various clinical MRSA strains, and has low biological toxicity and high yield, and is expected to be further developed into a potential clinical antibacterial drug. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 This is the dynamic bactericidal curve of compound 14.

[0036] Figure 2 The blood routine test and blood biochemical indexes of compound 14 in vivo (subcutaneous injection).

[0037] Figure 3 The changes in bacterial load on mouse skin after compound 14 (subcutaneous injection).

[0038] Figure 4 The blood routine test and blood biochemical indexes of compound 14 in vivo (intraperitoneal injection).

[0039] Figure 5 Changes in bacterial load in organs of mice injected with compound 14 at lethal and sublethal concentrations (intraperitoneal injection).

[0040] Figure 6 Changes in bacterial burden in mice organs after administration of compound 14 (intraperitoneal injection).

[0041] Figure 7 is the survival rate of mice injected with compound 14 (intraperitoneal injection)

[0042] Figure 8 For compound 14 1 H-NMR spectrum.

[0043] Fig. 9 For compound 14 13 C-NMR spectrum. DETAILED DESCRIPTION

[0044] The present invention is further described in detail below through examples.

[0045] Example 1 Preparation of Intermediate 2

[0046] Add rutaecarpine (1mmol) to a 25mL round-bottom flask, dissolve it in acetone as a reaction solvent, add dibromoalkane (12mmol) and inorganic base potassium carbonate (7mmol), and stir under reflux at 50°C in an oil bath for 72h. After monitoring the completion of the reaction of rutaecarpine by thin layer chromatography (TLC), the acetone in the reaction solution is concentrated under reduced pressure, extracted with dichloromethane, and dried over anhydrous sodium sulfate. The concentrate is separated and purified by silica gel column chromatography using a developing solvent of petroleum ether: ethyl acetate = 5:1 to obtain a white solid intermediate 2.

[0047] Example 2 Preparation of Intermediate 3

[0048] After the intermediate 2 (1 mmol) was dissolved in an appropriate amount of acetone, 4-mercaptopyridine (1 mmol), the catalyst inorganic base potassium carbonate (1.5 mmol), and the catalyst potassium iodide (0.02 mmol) were added, and the mixture was refluxed and stirred at 50°C in an oil bath for 12 h. After monitoring the completion of the reaction of the intermediate 2 by thin layer chromatography (TLC), the acetone in the reaction solution was concentrated under reduced pressure, extracted with dichloromethane, and dried over anhydrous sodium sulfate. The concentrate was separated and purified by silica gel column chromatography using a developing solvent of dichloromethane: methanol: acetone = 35:1:1 to obtain a yellow solid intermediate 3.

[0049] Example 3 Preparation of chloroacetamide

[0050] Add aliphatic amine / aromatic amine (1mmol) containing different substituents to a 25mL round-bottom flask, add an appropriate amount of dichloromethane as the reaction solvent, and place it under ice bath conditions (0℃). Slowly add chloroacetyl chloride (1.5mmol) dropwise within 10min, stir for 2-3min, then quickly add triethylamine (1.5mmol), and react for 5h under nitrogen protection. After monitoring the reaction by thin layer chromatography (TLC), extract with dichloromethane and dry with anhydrous sodium sulfate. The concentrate is separated and purified by silica gel column chromatography with dichloromethane: methanol = 10:1 to obtain brown oily chloroacetamide containing different substituents.

[0051] Example 4 Preparation of Intermediate 4

[0052] After the intermediate 3 (1 mmol) was dissolved in an appropriate amount of anhydrous acetonitrile as the reaction solvent, chloroacetamide (2 mmol) was added, and the mixture was placed in an oil bath at 80°C under nitrogen protection and refluxed for 12 h. After the reaction was completed by monitoring by thin layer chromatography (TLC), silica gel column chromatography was performed with dichloromethane:methanol = 10:1 to obtain a light yellow solid Evodia rutaecarpa alkaloid-quaternary ammonium salt compound.

[0053] Example 5 Compound 1

[0054] The intermediate 3 (1 mmol) was dissolved in anhydrous acetonitrile, chloroacetamide (2 mmol) was added, and the mixture was placed in an oil bath at 80°C for reflux and stirred for 12 h under nitrogen protection. After the reaction was completed by monitoring by thin layer chromatography (TLC), silica gel column chromatography was performed with dichloromethane:methanol = 10:1 to obtain a light yellow solid Evodia rutaecarpa alkaloid-quaternary ammonium salt compound.

[0055] The physicochemical properties of compound 1 are as follows:

[0056] 1) White solid;

[0057] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0058] With CDCl3 as solvent, the peaks are attributed to: 1H NMR(400MHz CDCl3)δ:8.73(d,J=6.48Hz,2H,-Ph),8.31(dd,J=7.92,1.08Hz,1H,-Ph),7.75(t,J=6.96Hz,1H,-Ph),7.66(d,J=5.76Hz,2H,-Ph),7.42-7.47(m,4 H,-Ph),7.39(d,J=7.68Hz,1H,-Ph),7.21(t,J=7.16Hz,1H,-Ph),6.04(s,2H,-CH2-),5.06(t,J=6.72Hz,2H,-CH2-),4.54(t,J=6.72Hz,2H,-CH2-) ,3.46(t,J=7.64Hz,2H,-CH2-),3.30(t,J=7.6Hz,2H,-S-CH2-),3.18-3. 24(m,4H,-CH2-),2.43-2.50(m,2H,-CH2-),1.61-1.69(m,2H,-CH2-),1.4 HRMS(ESI)C 36 H 42 ClN5O2S[M-Cl] + calcd=608.3054; found=608.3061.

[0059] Example 6 Compound 2

[0060] Compound 2 was synthesized by the method described in Example 5. The physicochemical properties of compound 2 are as follows:

[0061] 1) White solid;

[0062] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0063] With CDCl3 as solvent, the peaks are attributed to: 1HNMR(400MHz CDCl3)δ: 8.74 (d, J=5.92Hz, 2H, -Ph), 8.31 (d, J=7.32Hz, 1H, -Ph), 7.75 (t, J=7.08Hz, 1H, -Ph), 7.66 (dd, J=7.84, 3.2Hz, 2H, -Ph), 7. 41-7.45(m,4H,-Ph),7.39(d,J=8.08Hz,1H,-Ph),7.20(t,J=7.2Hz,1H,-Ph),6.06(s,2H,-CH2-),5.06(t,J=6.64Hz,2H,-CH2-),4.53 (t,J=6.72Hz,2H,-CH2-),3.43(t,J=7.52Hz,2H,-CH2-),3.27(t,J=7.68Hz,2H,-S-CH2-),3.18-3.24(m,4H,-CH2-),2.43-2.48(m,2H ,-CH2-),1.68-1.76(m,2H,-CH2-),1.52-1.60(m,2H,-CH2-),0.99(t,J=7.28Hz,3H,-CH3),0.87(t,J=7.32Hz,3H,-CH3); HRMS(ESI)C 34 H 38 ClN5O2S[M-Cl] + calcd=580.2741; found=580.2745.

[0064] Example 7 Compound 3

[0065] Compound 3 was synthesized by the method described in Example 5. The physicochemical properties of compound 3 are as follows:

[0066] 1) White solid;

[0067] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0068] With CDCl3 as solvent, the peaks are attributed to: 1HNMR(400MHz CDCl3)δ:9.28(t,J=5.44Hz,1H,-Ph),8.81(d,J=6.76Hz,2H,-Ph),8.32(dd,J=7.92,1.16Hz,1H,-NH-),7.76(t,J=6.88Hz ,1H,-Ph),7.68(dd,J=12.48,7.96Hz,2H,-Ph),7.43-7.48(m,4H,-Ph),7.40(td,J=7.68,0.84Hz,1H,-Ph),7.22(td,J=8.8 ,7.8Hz,1H,-Ph),5.59(s,2H,-CH2-),5.09(t,J=6.6Hz,2H,-CH2-),4.54(t,J=6.76Hz,2H,-CH2-),3.19-3.25(m,6H,-CH2- ),2.45-2.52(m,2H,-CH2-),1.51-1.58(m,2H,-CH2-),1.32-1.37(m,2H,-CH2-),0.88(t,J=7.32Hz,3H,-CH3); HRMS(ESI)C 32 H 34 ClN5O2S[M-Cl] + calcd=552.2428; found=552.2435.

[0069] Example 8 Compound 4

[0070] Compound 4 was synthesized by the method described in Example 5. The physicochemical properties of compound 4 are as follows:

[0071] 1) Yellow solid;

[0072] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0073] DMSO was used as solvent, and the peaks were assigned as follows: 1HNMR(400MHz DMSO)δ:8.63(t,J=5.52Hz,1H,-Ph),8.59(d,J=7.04Hz,2H,-Ph),8.17(dd,J=7.88,1.08Hz,1H,-NH-),7.98(d,J=7.08Hz,2H,-Ph),7.82(td,J =8.28,1.44Hz,1H,-Ph),7.69-7.33(m,3H,-Ph),7.49(t,J=7.84Hz,1H,-Ph),7.38(t,J=7.4Hz,1H,-Ph),7.17(t,J=7.56Hz,1H,-Ph),5.23(s,2 H,-CH2-),5.02(t,J=6.92Hz,2H,-CH2-),4.43(t,J=6.72Hz,2H,-CH2-),3.45(t,J=6.96Hz,2H,-CH2-),3.17(t,J=6.64Hz,2H,-CH2-),3.10(t, J=5.84Hz,2H,-CH2-),2.29-2.32(m,2H,-CH2-),1.42-1.46(m,2H,-CH2-),1.26-1.29(m,4H,-CH2-),0.87(t,J=6.80Hz,3H,-CH3); HRMS(ESI)C 33 H 36 ClN5O2S[M-Cl] + calcd=566.2584; found=566.2589.

[0074] Example 9 Compound 5

[0075] Compound 5 was synthesized by the method described in Example 5. The physicochemical properties of Compound 5 are as follows:

[0076] 1) White solid;

[0077] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0078] DMSO was used as solvent, and the peaks were assigned as follows: 1HNMR(400MHz DMSO)δ:8.67(t,J=5.40Hz,1H,-Ph),8.60(d,J=7.08Hz,2H,-Ph),8.16(dd,J=7.96,1.2Hz,1H,-NH-),7.98(d,J=7.16Hz,2H,-Ph),7.82(td, J=6.84,1.48Hz,1H,-Ph),7.69-7.73(m,3H,-Ph),7.49(td,J=7.04,0.96Hz,1H,-Ph),7.38(td,J=7.32,0.8Hz,1H,-Ph),7.17(t,J=7.64Hz, 1H,-Ph),5.24(s,2H,-CH2-),5.02(t,J=6.96Hz,2H,-CH2-),4.43(t,J=6.72Hz,2H,-CH2-),3.46(t,J=6.96Hz,2H,-CH2-),3.17(t,J=6.72H z,2H,-CH2-),3.09(q,J=6.84Hz,2H,-CH2-),2.27-2.32(m,2H,-CH2-),1.42-1.51(m,2H,-CH2-),0.88(t,J=7.36Hz,3H,-CH3); HRMS(ESI)C 31 H 32 ClN5O2S[M-Cl] + calcd=538.2271; found=538.2278.

[0079] Example 10 Compound 6

[0080] Compound 6 was synthesized by the method described in Example 5. The physicochemical properties of compound 6 are as follows:

[0081] 1) White solid;

[0082] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0083] DMSO was used as solvent, and the peaks were assigned as follows: 1HNMR(400MHz DMSO)δ:8.57(d,J=7.08Hz,2H,-Ph),8.16(dd,J=7.92,1.2Hz,1H,-Ph),8.01(d,J=7.12Hz,2H,-Ph),7.82(td,J=8.32,1.44H z,1H,-Ph),7.72(t,J=5.12Hz,3H,-Ph),7.50(td,J=6.24,0.8Hz,1H,-Ph),7.38(t,J=7.12Hz,1H,-Ph),7.18(t,J=7.68Hz,1 H,-Ph),5.60(s,2H,-CH2-),5.03(t,J=6.84Hz,2H,-CH2-),4.43(t,J=6.72Hz,2H,-CH2-),3.70(t,J=4.28Hz,2H,-CH2-),3. 61(t,J=4.32Hz,2H,-CH2-),3.44-3.48(m,6H,-O-CH2-,-N-CH2-),3.17(t,J=6.72Hz,2H,-CH2-),2.28-2.35(m,2H,-CH2-), 13 C NMR(100MHz DMSO)δ:163.5,162.9,160.5,146.7,145.2,143.7,139.3,126.3,125.9,123.7,122.0,120.3,119.6,18.9; HRMS(ESI)C 32 H 32 ClN5O3S[M-Cl] + calcd=566.2220; found=566.2228.

[0084] Example 11 Compound 7

[0085] Compound 7 was synthesized by the method described in Example 5. The physicochemical properties of compound 7 are as follows:

[0086] 1) Yellow solid;

[0087] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0088] DMSO was used as solvent, and the peaks were assigned as follows: 1HNMR(400MHz DMSO)δ:8.58(d,J=6.88Hz,2H,-Ph),8.16(d,J=6.96Hz,1H,-Ph),8.01(d,J=6.92 Hz,2H,-Ph),7.82(t,J=8.16Hz,1H,-Ph),7.72(t,J=8.32Hz,3H,-Ph),7.49(t,J= 7.76Hz,1H,-Ph),7.38(t,J=7.40Hz,1H,-Ph),7.18(t,J=7.64Hz,1H,-Ph),5.59( s,2H,-CH2-),5.03(t,J=6.84Hz,2H,-CH2-),4.43(t,J=6.68Hz,2H,-CH2-),3.71 -3.74(m,4H,-N-CH2-),3.46(t,J=6.84Hz,2H,-CH2-),3.17(t,J=6.68Hz,2H,-CH2 -),2.79-2.81(m,2H,-CH2-),2.59-2.62(m,2H,-CH2-),2.29-2.33(m,2H,-CH2-), 13 C NMR(100MHz DMSO)δ:163.46,162.93,160.51,146.7,145.3,143.8,139.3,126.3,126.0,123.8,122.0,120.3,119.6,18.9; HRMS(ESI)C 32 H 32 ClN5O2S2[M-Cl] + calcd=582.1992; found=582.1993.

[0089] Example 12 Compound 8

[0090] Compound 8 was synthesized by the method described in Example 5. The physicochemical properties of compound 8 are as follows:

[0091] 1) Yellow solid;

[0092] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0093] DMSO was used as solvent, and the peaks were assigned as follows: 1HNMR(400MHz DMSO)δ:8.57(d,J=6.96Hz,2H,-Ph),8.17(d,J=7.20Hz,1H,-Ph),8.01(d,J=7.00H z,2H,-Ph),7.83(t,J=7.04Hz,1H,-Ph),7.72(td,J=4.64,3.96Hz,3H,-Ph),7.50(t ,J=7.52Hz,1H,-Ph),7.38(t,J=7.32Hz,1H,-Ph),7.18(t,J=7.60Hz,1H,-Ph),5.4 8(s,2H,-CH2-),5.02(t,J=6.76Hz,2H,-CH2-),4.43(t,J=6.68Hz,2H,-CH2-),3.43 HRMS(ESI)C 32 H 32 ClN5O2S[M-Cl] + calcd=550.2271; found=550.2280.

[0094] Example 13 Compound 9

[0095] Compound 9 was synthesized by the method described in Example 5. The physicochemical properties of compound 9 are as follows:

[0096] 1) Yellow solid;

[0097] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0098] DMSO was used as solvent, and the peaks were assigned as follows: 1HNMR(400MHz DMSO)δ:8.58(d,J=6.76Hz,2H,-Ph),8.16(dd,J=7.88,1.08Hz,1H,-Ph),8.00(d,J=6.92Hz,2H,-Ph),7.82(td,J=8.32,1 .44Hz,1H,-Ph),7.71(t,J=6.64Hz,3H,-Ph),7.56-7.58(m,4H,-Ph),7.49(td,J=8.00,0.96Hz,2H,-Ph),7.38(t,J=7.12H z,1H,-Ph),7.17(t,J=7.60Hz,1H,-Ph),5.16(s,2H,-CH2-),5.02(t,J=6.92Hz,2H,-CH2-),4.42(t,J=6.72Hz,2H,-CH2- ),3.46(t,J=7.00Hz,2H,-CH2-),3.23(s,3H,-CH3),3.17(t,J=6.72Hz,2H,-CH2-),2.27-2.34(m,2H,-CH2-); HRMS(ESI)C 35 H 32 ClN5O2S[M-Cl] + calcd=586.2271; found=586.2279.

[0099] Example 14 Compound 10

[0100] Compound 10 was synthesized by the method described in Example 5. The physicochemical properties of compound 10 are as follows:

[0101] 1) Yellow solid;

[0102] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0103] DMSO was used as solvent, and the peaks were assigned as follows: 1HNMR(400MHz DMSO)δ:8.62(d,J=7.20Hz,1H,-Ph),8.58(d,J=7.08Hz,2H,-Ph),8.17(dd,J=7.96,1.20Hz,1H,-NH-),7.97(d,J=7.12Hz,2H,-Ph),7.82(t,J= 6.84Hz,1H,-Ph),7.69-7.73(m,3H,-Ph),7.49(t,J=8.00Hz,1H,-Ph),7.38(t,J=7.24Hz,1H,-Ph),7.17(t,J=7.40Hz,1H,-Ph),5.16(s,2H,-CH 2-),5.02(t,J=6.64Hz,2H,-CH2-),4.43(t,J=6.80Hz,2H,-CH2-),3.96-4.05(m,1H,-CH-),3.45(t,J=6.84Hz,2H,-CH2-),3.17(t,J=6.76Hz, 2H,-CH2-),2.29-2.34(m,2H,-CH2-),1.70-1.80(m,2H,-CH2-),1.64-1.68(m,2H,-CH2-),1.50-1.54(m,2H,-CH2-),1.41-1.47(m,2H,-CH2-), 13 C NMR(100MHz DMSO)δ:163.73,160.50,146.78,145.31,143.74,139.36,126.36,125.99,123.79,121.99,120.37,119.65,54.87,32.17,23.37; HRMS(ESI)C 33 H 34 ClN5O2S[M-Cl] + calcd=564.2428; found=564.2430.

[0104] Example 15 Compound 11

[0105] Compound 11 was synthesized by the method described in Example 5. The physicochemical properties of compound 11 are as follows:

[0106] 1) Yellow solid;

[0107] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0108] DMSO was used as solvent, and the peaks were assigned as follows: 1HNMR (400MHz DMSO) δ: 8.60 (d, J = 6.80Hz, 2H, -Ph), 8.16 (d, J = 7.96Hz, 1H, -Ph), 7.93 (d, J = 7. 00Hz,2H,-Ph),7.78(t,J=6.96Hz,1H,-Ph),7.69(t,J=8.72Hz,2H,-Ph),7.62(d ,J=8.08Hz,1H,-Ph),7.47(t,J=7.12Hz,1H,-Ph),7.37(t,J=7.28Hz,1H,-Ph), 7.16(t,J=7.24Hz,1H,-Ph),5.61(s,2H,-CH2-),4.92(t,J=7.00Hz,2H,-CH2-), 4.43(t,J=6.72Hz,2H,-CH2-),3.27-3.37(m,6H,-CH2-),3.17(t,J=6.60Hz,2H ,-CH2-),2.00-2.07(m,2H,-CH2-),1.79-1.86(m,2H,-CH2-),1.60-1.68(m,2H, -CH2-),1.43-1.51(m,2H,-CH2-),1.33-1.37(m,2H,-CH2-),1.24-1.27(m,2H, -CH2-),0.96(t,J=7.24Hz,3H,-CH3),0.87(t,J=7.28Hz,3H,-CH3); HRMS(ESI)C 37 H 44 ClN5O2S[M-Cl] + calcd=622.3210; found=622.3214.

[0109] Example 16 Compound 12

[0110] Compound 12 was synthesized by the method described in Example 5. The physicochemical properties of compound 12 are as follows:

[0111] 1) Yellow solid;

[0112] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0113] DMSO was used as solvent, and the peaks were assigned as follows: 1HNMR(400MHz DMSO)δ:8.60(d,J=7.08Hz,2H,-Ph),8.17(dd,J=7.96,1.16Hz,1H,-Ph),7.93(d,J=7.12Hz,2H,-Ph),7.78(t,J=6.88Hz,1H,-Ph),7.69(t,J=8.76Hz ,2H,-Ph),7.62(d,J=7.88Hz,1H,-Ph),7.47(t,J=7.92Hz,1H,-Ph),7.37(t,J=7.24Hz,1H,-Ph),7.16(t,J=7.44Hz,1H,-Ph),5.60(s,2H,-CH2-),4. 92(t,J=7.12Hz,2H,-N-CH2-),4.43(t,J=6.68Hz,2H,-N-CH2-),3.24-3.3 5(m,6H,-N-CH2-,-CH2-),3.17(t,J=6.64Hz,2H,-CH2-),2.00-2.07(m,2H ,-CH2-),1.79-1.86(m,2H,-CH2-),1.63-1.72(m,2H,-CH2-),1.46-1.55( m,2H,-CH2-),0.94(t,J=7.28Hz,3H,-CH3),0.83(t,J=7.36Hz,3H,-CH3), 13 C NMR(100MHz DMSO)δ:164.24,162.85,160.51,146.79,145.31,143.89,139.46,134.40,126.38,125.98,123.69,122 .01,120.35,119.56,48.10.47.46,43.82,40.55,30.27,24.73,21.28,20.34,18.92,11.12; HRMS(ESI)C 35 H 40 ClN5O2S[M-Cl] + calcd=594.2897; found=594.2902.

[0114] Example 17 Compound 13

[0115] Compound 13 was synthesized by the method described in Example 5. The physicochemical properties of compound 13 are as follows:

[0116] 1) Yellow solid;

[0117] 2) The NMR spectrum of the compound ( 1 H / 13C NMR, 400MHz) characteristics:

[0118] DMSO was used as solvent, and the peaks were assigned as follows: 1 HNMR(400MHz DMSO)δ:8.71(t,J=5.40Hz,1H,-Ph),8.60(d,J=7.12Hz,2H,-Ph),8.16(dd,J=7.96,1.28Hz,1H,-NH-),7.91(d,J=7.16Hz,2H,-Ph),7.78(t,J=6.8 8Hz,1H,-Ph),7.68(t,J=8.92Hz,2H,-Ph),7.62(d,J=7.84Hz,1H,-Ph),7.47(t,J=7.04Hz,1H,-Ph),7.37(t,J=7.20Hz,1H,-Ph),7.16(t,J=7.32H z,1H,-Ph),5.27(s,2H,-CH2-),4.91(t,J=7.08Hz,2H,-CH2-),4.42(t,J=6.72Hz,2H,-CH2-),3.33-3.34(m,2H,-CH2-),3.10-3.18(m,4H,-S-CH2 -,-CH2-),1.99-2.06(m,2H,-CH2-),1.75-1.85(m,2H,-CH2-),1.40-1.47(m,2H,-CH2-),1.29-1.36(m,2H,-CH2-),0.88(t,J=7.28Hz,3H,-CH3), 13 C NMR(100MHz DMSO)δ:164.35,162.74,160.51,146.78,145.30,143.62,139.45,134.39,126.94,126.37,125.97,123. 68,122.06,120.34,119.56,110.81,59.92,43.82,40.55,30.94,30.23,24.73,19.47,18.92; HRMS(ESI)C 33 H 36 ClN5O2S[M-Cl] + calcd=566.2584; found=566.2595.

[0119] Example 18 Compound 14

[0120] Compound 14 was synthesized by the method described in Example 5. The physicochemical properties of compound 14 are as follows:

[0121] 1) Yellow solid;

[0122] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0123] DMSO was used as solvent, and the peaks were assigned as follows: 1 HNMR(400MHz DMSO)δ:8.71(t,J=5.44Hz,1H,-Ph),8.60(d,J=7.16Hz,2H,-Ph),8.16(dd,J=7.96,1.24Hz,1H,-NH-),7.91(d,J=7.16Hz,2H,-Ph),7.78(td,J=8.36, 1.48Hz,1H,-Ph),7.68(t,J=9.00Hz,2H,-Ph),7.62(d,J=7.88Hz,1H,-Ph),7.47(td,J=8.00,0.96Hz,1H,-Ph),7.37(t,J=7.24Hz,1H,-Ph),7.16(t,J =7.56Hz,1H,-Ph),5.26(s,2H,-CH2-),4.91(t,J=7.08Hz,2H,-CH2-),4.4 2(t,J=6.68Hz,2H,-CH2-),3.35-3.36(m,2H,-CH2-),3.12-3.18(m,4H,-S- CH2-,-CH2-),1.99-2.06(m,2H,-CH2-),1.75-1.85(m,2H,-CH2-),1.42-1 .47(m,2H,-CH2-),1.26-1.29(m,4H,-CH2-),0.87(t,J=6.76Hz,3H,-CH3), 13 C NMR(100MHz DMSO)δ:164.35,162.72,160.50,146.78,145.30,143.67,139.44,126.96,126.37,125.96,123.67,1 22.04,120.30,119.55,110.83,59.90,43.81,40.54,30.21,28.51,24.72,21.76,18.91; HRMS(ESI)C 34 H 38 ClN5O2S[M-Cl] + calcd=580.2741; found=580.2745.

[0124] Example 19 Compound 15

[0125] Compound 15 was synthesized by the method described in Example 5. The physicochemical properties of compound 15 are as follows:

[0126] 1) Yellow solid;

[0127] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0128] DMSO was used as solvent, and the peaks were assigned as follows: 1 HNMR(400MHz DMSO)δ:8.73(t,J=4.96Hz,1H,-Ph),8.61(d,J=7.04Hz,2H,-Ph),8.16(dd,J=7.88,1.16Hz,1H,-NH-),7.91(d,J=7.12Hz,2H,-Ph),7.78(t,J=6.9 2Hz,1H,-Ph),7.69(t,J=8.88Hz,2H,-Ph),7.62(d,J=7.96Hz,1H,-Ph),7.47(t,J=7.08Hz,1H,-Ph),7.37(t,J=7.24Hz,1H,-Ph),7.16(t,J=7.28H z,1H,-Ph),5.27(s,2H,-CH2-),4.91(t,J=7.00Hz,2H,-CH2-),4.42(t,J=6.76Hz,2H,-CH2-),3.35-3.36(m,2H,-CH2-),3.17(t,J=6.68Hz,2H,,- CH2-),3.10(q,J=6.76Hz,2H,-CH2-),1.99-2.06(m,2H,-CH2-),1.78-1.85(m,2H,-CH2-),1.42-1.51(m,2H,-CH2-),0.88(t,J=7.36Hz,3H,-CH3), 13 C NMR(100MHz DMSO)δ:164.40,162.75,160.51,146.78,145.31,143.66,139.46,134.40,126.37,125.9 8,123.68,122.05,120.35,119.56,40.82,40.55,30.23,24.73,22.13,18.22; HRMS(ESI)C 32 H 34 ClN5O2S[M-Cl] + calcd=552.2428; found=552.2431.

[0129] Example 20 Compound 16

[0130] Compound 16 was synthesized by the method described in Example 5. The physicochemical properties of compound 16 are as follows:

[0131] 1) Yellow solid;

[0132] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0133] DMSO was used as solvent, and the peaks were assigned as follows: 1 HNMR(400MHz DMSO)δ:8.57(d,J=6.84Hz,2H,-Ph),8.16(d,J=7.96Hz,1H,-Ph),7.93(d,J=6.88Hz,2H,-Ph),7.78(t,J=7.20Hz,1H,-Ph),7.69(t,J=8.48H z,2H,-Ph),7.62(d,J=8.08Hz,1H,-Ph),7.48(t,J=7.44Hz,1H,-Ph),7.37(t,J=7.40Hz,1H,-Ph),7.16(t,J=7.48Hz,1H,-Ph),5.63(s,2H,- CH2-),4.92(t,J=6.96Hz,2H,-CH2-),4.43(t,J=6.64Hz,2H,-CH2-),3.71(t,J=4.48Hz,2H,-CH2-),3.61(t,J=4.16Hz,2H,-CH2-),3.47-3. 49(m,4H,-CH2-),3.33-3.37(m,2H,-CH2-),3.17(t,J=6.68Hz,2H,-CH2-),2.00-2.07(m,2H,-CH2-),1.80-1.85(m,2H,-CH2-); HRMS(ESI)C 33 H 34 ClN5O3S[M-Cl] + calcd=580.2377; found=580.2385.

[0134] Example 21 Compound 17

[0135] Compound 17 was synthesized by the method described in Example 5. The physicochemical properties of compound 17 are as follows:

[0136] 1) Yellow solid;

[0137] 2) The NMR spectrum of the compound ( 1 H / 13C NMR, 400MHz) characteristics:

[0138] DMSO was used as solvent, and the peaks were assigned as follows: 1 HNMR(400MHz DMSO)δ:8.57(d,J=6.68Hz,2H,-Ph),8.16(dd,J=7.84,1.04Hz,1H,-Ph),7.94(d,J=7.04Hz,2H,-Ph),7.78(t,J=6.88Hz,1H,-Ph),7.69( t,J=8.12Hz,2H,-Ph),7.62(d,J=7.96Hz,1H,-Ph),7.48(t,J=7.84Hz,1H,-Ph),7.37(t,J=7.68Hz,1H,-Ph),7.16(t,J=7.48Hz,1H,-Ph) ,5.61(s,2H,-CH2-),4.92(t,J=7.04Hz,2H,-CH2-),4.43(t,J=6.64Hz,2H,-CH2-),3.72-3.75(m,4H,-CH2-),3.34-3.37(m,2H,-CH2-), 3.17(t,J=6.68Hz,2H,-CH2-),2.79-2.83(m,2H,-CH2-),2.61-2.63(m,2H,-CH2-),1.98-2.05(m,2H,-CH2-),1.79-1.86(m,2H,-CH2-), 13 C NMR(100MHzDMSO)δ:163.53,162.95,160.51,146.77,145.30,143.78,139.44,134.40,12 6.37,125.98,123.67,122.09,120.34,120.28,119.57,110.82,24.67,18.92; HRMS(ESI)C 33 H 34 ClN5O2S2[M-Cl] + calcd=596.2148; found=596.2158.

[0139] Example 22 Compound 18

[0140] Compound 18 was synthesized by the method described in Example 5. The physicochemical properties of compound 18 are as follows:

[0141] 1) Yellow solid;

[0142] 2) The NMR spectrum of the compound ( 1 H / 13C NMR, 400MHz) characteristics:

[0143] DMSO was used as solvent, and the peaks were assigned as follows: 1 HNMR(400MHz DMSO)δ:8.57(d,J=6.84Hz,2H,-Ph),8.16(dd,J=7.92,1.12Hz,1H,-Ph),7.93(d,J=7.12Hz,2H,-Ph),7.78(t,J=6.88Hz,1H, -Ph),7.69(t,J=8.92Hz,2H,-Ph),7.62(d,J=8.04Hz,1H,-Ph),7.48(t,J=7.88Hz,1H,-Ph),7.37(t,J=7.92Hz,1H,-Ph),7.1 6(t,J=7.56Hz,1H,-Ph),5.51(s,2H,-CH2-),4.92(t,J=6.96Hz,2H,-CH2-),4.43(t,J=6.72Hz,2H,-CH2-),3.51(t,J=6.72H z,2H,-CH2-),3.36-3.37(m,4H,-CH2-),3.17(t,J=6.68Hz,2H,-CH2-),1.94-2.05(m,4H,-CH2-),1.81-1.86(m,4H,-CH2-), 13 C NMR(100MHz DMSO)δ:162.89,162.84,160.51,146.78,145.30,143.70,139.45,134.40,126.37,125.97,1 23.68,122.07,120.34,119.56,46.06,40.55,30.25,25.46,24.70,23.72,18.91; HRMS(ESI)C 33 H 34 ClN5O2S[M-Cl] + calcd=564.2428; found=564.2437.

[0144] Example 23 Compound 19

[0145] Compound 19 was synthesized by the method described in Example 5. The physicochemical properties of compound 19 are as follows:

[0146] 1) Yellow solid;

[0147] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0148] DMSO was used as solvent, and the peaks were assigned as follows: 1 HNMR(400MHz DMSO)δ:8.57(d,J=6.72Hz,2H,-Ph),8.16(dd,J=7.88,1.04Hz,1H,-Ph),7.92(d,J=6.76Hz,2H,-Ph),7.77(t,J=6.96Hz, 1H,-Ph),7.68(t,J=8.80Hz,2H,-Ph),7.57-7.62(m,5H,-Ph),7.47(t,J=7.12Hz,2H,-Ph),7.37(t,J=7.36Hz,1H,-Ph),7 .16(t,J=7.28Hz,1H,-Ph),5.17(s,2H,-CH2-),4.91(t,J=7.08Hz,2H,-CH2-),4.42(t,J=6.72Hz,2H,-CH2-),3.37-3.38 (m,2H,-CH2-),3.24(s,3H,-N-CH3),3.17(t,J=6.72Hz,2H,-CH2-),1.99-2.07(m,2H,-CH2-),1.78-1.86(m,2H,-CH2-), 13 C NMR(100MHz DMSO)δ:164.34,162.93,160.51,146.78,145.30,143.88,139.44,134.40,130.00,127.64,12 6.37,125.96,123.67,121.84,120.33,119.55,40.55,37.43,30.25,24.72,18.92; HRMS(ESI)C 36 H 34 ClN5O2S[M-Cl] + calcd=600.2428; found=600.2436.

[0149] Example 24 Compound 20

[0150] Compound 20 was synthesized by the method described in Example 5. The physicochemical properties of compound 20 are as follows:

[0151] 1) Yellow solid;

[0152] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0153] DMSO was used as solvent, and the peaks were assigned as follows: 1HNMR(400MHz DMSO)δ:8.69(d,J=6.84Hz,1H,-Ph),8.59(d,J=7.00Hz,2H,-Ph),8.17(dd,J=7.92,1.12Hz,1H,-NH-),7.90(d,J=7.04Hz,2H,-Ph),7.78(t,J=6 .96Hz,1H,-Ph),7.69(t,J=8.00Hz,2H,-Ph),7.62(d,J=8.04Hz,1H,-Ph),7.48(t,J=7.12Hz,1H,-Ph),7.37(t,J=7.32Hz,1H,-Ph),7.16(t,J=7 .32Hz,1H,-Ph),5.20(s,2H,-CH2-),4.91(t,J=7.08Hz,2H,-CH2-),4.42(t,J=6.76Hz,2H,-CH2-),3.98-4.06(m,1H,-CH-),3.33-3.35(m,2H,- CH2-),3.17(t,J=6.68Hz,2H,-CH2-),1.99-2.06(m,2H,-CH2-),1.80-1.85(m,4H,-CH2-),1.65-1.68(m,2H,-CH2-),1.43-1.55(m,4H,-CH2-); 13 C NMR(100MHz DMSO)δ:163.79,162.69,160.51,146.78,145.30,143.64,139.44,134.42,126.37,125.96,123.6 7,122.01,120.34,119.56,110.84,59.93,40.55,32.17,30.21,24.73,23.37,18.91; HRMS(ESI)C 34 H 36 ClN5O2S[M-Cl] + calcd=578.2584; found=578.2589.

[0154] Example 25 Compound 21

[0155] Compound 21 was synthesized by the method described in Example 5. The physicochemical properties of compound 21 are as follows:

[0156] 1) Yellow solid;

[0157] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0158] DMSO was used as solvent, and the peaks were assigned as follows: 1 HNMR(400MHz DMSO)δ:8.58(d,J=6.76Hz,2H,-Ph),8.16(d,J=7.92Hz,1H,-Ph),7.92(d,J=6.96Hz,2H,-Ph),7.79(t,J=7.00Hz,1H,-Ph),7.62-7.71(m,3H,-Ph), 7.48(t,J=7.16Hz,1H,-Ph),7.36(t,J=7.24Hz,1H,-Ph),7.15(t,J=7.32Hz,1H,-Ph),5.58(s,2H,-CH2-),4.88(t,J=7.24Hz,2H,-CH2-),4.43(t,J= 6.80Hz,2H,-CH2-),3.24-3.29(m,6H,-CH2-),3.16(t,J=6.76Hz,2H,-CH 2-),1.86-1.94(m,2H,-CH2-),1.74-1.82(m,2H,-CH2-),1.56-1.67(m,4H ,-CH2-),1.42-1.49(m,2H,-CH2-),1.35-1.38(m,2H,-CH2-),1.22-1.26( m,2H,-CH2-),0.95(t,J=7.24Hz,3H,-CH3),0.86(t,J=7.24Hz,3H,-CH3), 13 C NMR(100MHz DMSO)δ:164.11,162.92,160.53,146.81,145.35,143.89,139.42,134.39,126.39,125.98,125.12,123.68,121.97 ,120.37,119.46,46.29,45.58,44.20,40.54,30.42,30.11,29.24,27.26,25.35,19.54,18.94,13.66; HRMS(ESI)C 38 H 46 ClN5O2S[M-Cl] + calcd=636.3367; found=636.3370.

[0159] Example 26 Compound 22

[0160] Compound 22 was synthesized by the method described in Example 5. The physicochemical properties of compound 22 are as follows:

[0161] 1) Yellow solid;

[0162] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0163] DMSO was used as solvent, and the peaks were assigned as follows: 1 HNMR(400MHz DMSO)δ:8.57(d,J=6.72Hz,2H,-Ph),8.17(d,J=7.00Hz,1H,-Ph),7.92(d,J=6.88Hz,2H,-Ph),7.80(t,J=8.24Hz,1H,-Ph),7.62-7.71(m,3H, -Ph),7.48(t,J=7.60Hz,1H,-Ph),7.36(t,J=7.72Hz,1H,-Ph),7.15(t,J=7.52Hz,1H,-Ph),5.57(s,2H,-CH2-),4.88(t,J=7.16Hz,2H,-CH2- ),4.43(t,J=6.60Hz,2H,-CH2-),3.24-3.26(m,6H,-CH2-),3.17(t,J=6.68Hz,2H,-CH2-),1.88-1.92(m,2H,-CH2-),1.76-1.80(m,2H,-CH2- ),1.64-1.71(m,2H,-CH2-),1.54-1.62(m,2H,-CH2-),1.45-1.52(m,2H,-CH2-),0.93(t,J=7.28Hz,3H,-CH3),0.82(t,J=7.28Hz,3H,-CH3); 13 C NMR(100MHz DMSO)δ:164.22,162.93,160.53,146.81,145.35,143.89,139.41,134.41,126.39,125.98,123.67,121.97,12 0.37,119.45,48.08,47.44,44.21,40.54,30.40,29.45,27.26,25.35,21.27,20.33,18.93,11.10; HRMS(ESI)C 36 H 42 ClN5O2S[M-Cl] + calcd=608.3054; found=608.3057.

[0164] Example 27 Compound 23

[0165] Compound 23 was synthesized by the method described in Example 5. The physicochemical properties of compound 23 are as follows:

[0166] 1) Yellow solid;

[0167] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0168] DMSO was used as solvent, and the peaks were assigned as follows: 1 HNMR(400MHz DMSO)δ:8.62(d,J=5.56Hz,1H,-Ph),8.58(d,J=6.92Hz,2H,-Ph),8.17(dd,J =7.92,1.12Hz,1H,-NH-),7.90(d,J=7.00Hz,2H,-Ph),7.79(td,J=8.32,1.3 6Hz,1H,-Ph)7.62-7.71(m,3H,-Ph),7.48(t,J=7.08Hz,1H,-Ph),7.36(t,J= 7.32Hz,1H,-Ph),7.15(t,J=7.44Hz,1H,-Ph),5.23(s,2H,-CH2-),4.87(t,J =7.24Hz,2H,-CH2-),4.42(t,J=6.72Hz,2H,-CH2-),3.25(t,J=7.20Hz,2H,- CH2-),3.16(t,J=6.72Hz,2H,-CH2-),3.12(q,J=6.76Hz,2H,-CH2-),1.86-1 .93(m,2H,-CH2-),1.73-1.81(m,2H,-CH2-),1.53-1.61(m,2H,-CH2-),1.39 -1.46(m,2H,-CH2-),1.28-1.32(m,2H,-CH2-),0.87(t,J=7.28Hz,3H,-CH3); 13 C NMR(100MHz DMSO)δ:164.34,162.84,160.52,146.80,145.35,143.65,139.41,134.40,126.39,125.98,123. 67,122.00,120.36,119.46,44.18,30.94,30.37,29.41,27.28,25.34,19.46,18.93; HRMS(ESI)C 34 H 38 ClN5O2S[M-Cl] + calcd=580.2741; found=580.2744.

[0169] Example 28 Compound 24

[0170] Compound 24 was synthesized by the method described in Example 5. The physicochemical properties of compound 24 are as follows:

[0171] 1) Yellow solid;

[0172] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0173] DMSO was used as solvent, and the peaks were assigned as follows: 1 HNMR(400MHz DMSO)δ:8.70(t,J=5.36Hz,1H,-Ph),8.59(d,J=6.96Hz,2H,-Ph),8.16(dd,J =7.84,1.04Hz,1H,-NH-),7.91(d,J=7.00Hz,2H,-Ph),7.79(t,J=6.96Hz,1H, -Ph)7.61-7.70(m,3H,-Ph),7.48(t,J=7.28Hz,1H,-Ph),7.35(t,J=7.36Hz, 1H,-Ph),7.15(t,J=7.44Hz,1H,-Ph),5.25(s,2H,-CH2-),4.87(t,J=7.20Hz, 2H,-CH2-),4.42(t,J=6.72Hz,2H,-CH2-),3.25(t,J=7.24Hz,2H,-CH2-),3. 16(t,J=6.72Hz,2H,-CH2-),3.11(q,J=6.76Hz,2H,-CH2-),1.86-1.93(m,2H, -CH2-),1.73-1.80(m,2H,-CH2-),1.53-1.61(m,2H,-CH2-),1.41-1.48(m,2H ,-CH2-),1.25-1.29(m,4H,-CH2-),0.86(t,J=6.72Hz,3H,-CH3); HRMS(ESI)C 35 H 40 ClN5O2S[M-Cl] + calcd=594.2897; found=594.2906.

[0174] Example 29 Compound 25

[0175] Compound 25 was synthesized by the method described in Example 5. The physicochemical properties of compound 25 are as follows:

[0176] 1) Yellow solid;

[0177] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0178] DMSO was used as solvent, and the peaks were assigned as follows: 1 HNMR(400MHz DMSO)δ:8.73(t,J=5.46Hz,1H,-Ph),8.60(d,J=6.88Hz,2H,-Ph),8.16(d,J=6.96Hz,1H,-NH-),7.91(d,J=7.04Hz,2H,-Ph),7.79(t,J=6.96Hz, 1H,-Ph),7.61-7.70(m,3H,-Ph),7.48(t,J=7.20Hz,1H,-Ph),7.35(t,J=7.28Hz,1H,-Ph),7.15(t,J=7.36Hz,1H,-Ph),5.26(s,2H,-CH2-),4.8 7(t,J=7.16Hz,2H,-CH2-),4.42(t,J=6.68Hz,2H,-CH2-),3.25(t,J=7.24Hz,2H,-CH2-),3.16(t,J=6.72Hz,2H,-CH2-),3.09(q,J=6.76Hz,2H, -CH2-),1.86-1.93(m,2H,-CH2-),1.73-1.80(m,2H,-CH2-),1.53-1.61(m,2H,-CH2-),1.41-1.50(m,2H,-CH2-),0.88(t,J=7.36Hz,3H,-CH3); 13 C NMR(100MHz DMSO)δ:164.40,162.82,160.52,146.80,145.34,143.64,139.40,134.40,126.85,126.38,125.97,125.11,123. 66,122.01,120.36,119.45,110.78,59.89,44.19,30.37,29.45,27.28,25.33,22.12,18.93,11.41; HRMS(ESI)C 33 H 36 ClN5O2S[M-Cl] + calcd=566.2584; found=566.2587.

[0179] Example 30 Compound 26

[0180] Compound 26 was synthesized by the method described in Example 5. The physicochemical properties of compound 26 are as follows:

[0181] 1) Yellow solid;

[0182] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0183] DMSO was used as solvent, and the peaks were assigned as follows: 1 HNMR(400MHz DMSO)δ:8.56(d,J=6.68Hz,2H,-Ph),8.16(d,J=6.84Hz,1H,-Ph),7.93(d,J=7.00Hz,2H,-Ph),7.79(t,J=6.92Hz,1H,-Ph),7.62-7.70(m ,3H,-Ph),7.48(t,J=7.20Hz,1H,-Ph),7.36(t,J=7.28Hz,1H,-Ph),7.15(t,J=7.40Hz,1H,-Ph),5.62(s,2H,-CH2-),4.87(t,J=7.24Hz, 2H,-CH2-),4.42(t,J=6.72Hz,2H,-CH2-),3.70(t,J=4.16Hz,2H,-CH2-),3.61(t,J=4.36Hz,2H,-CH2-),3.45-3.48(m,4H,-CH2-),3.26 (t,J=7.24Hz,2H,-CH2-),3.16(t,J=6.68Hz,2H,-CH2-),1.86-1.93(m,2H,-CH2-),1.74-1.81(m,2H,-CH2-),1.53-1.61(m,2H,-CH2-); 13 C NMR(100MHzDMSO)δ:163.70,163.07,160.52,146.80,145.35,143.77,139.41,134.40,126.88,126.39,125 .98,123.67,122.07,120.36,120.19,119.46,59.48,44.19,30.43,29.45,27.25,25.34,18.93; HRMS(ESI)C 34 H 36 ClN5O3S[M-Cl] + calcd=594.2533; found=594.2542.

[0184] Example 31 Compound 27

[0185] Compound 27 was synthesized by the method described in Example 5. The physicochemical properties of compound 27 are as follows:

[0186] 1) Yellow solid;

[0187] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0188] DMSO was used as solvent, and the peaks were assigned as follows: 1 HNMR(400MHz DMSO)δ:8.57(d,J=6.92Hz,2H,-Ph), 8.16(dd,J=7.92,1.12Hz,1H,-Ph), 7.94(d,J=7.08Hz,2H,-Ph), 7.79(td,J=8.32,1.36Hz,1H,-Ph), 7.62-7.71(m,3H,-Ph),7.48(t,J=7.16Hz,1H,-Ph),7.36(t,J=7.32Hz,1H,-Ph),7.15(t,J=7.36Hz,1H,-Ph),5.61(s,2H,-CH2-),4.88(t ,J=7.08Hz,2H,-CH2-),4.42(t,J=6.72Hz,2H,-CH2-),3.71-3.74(m,4H,-CH2-),3.26(t,J=7.28Hz,2H,-CH2-),3.16(t,J=6.72Hz,2H,-C H2-),2.75-2.82(m,2H,-CH2-),2.59-2.61(m,2H,-CH2-),1.86-1.93(m,2H,-CH2-),1.74-1.81(m,2H,-CH2-),1.54-1.61(m,2H,-CH2-); 13 C NMR(100MHzDMSO)δ:163.53,163.05,160.53,146.80,145.35,143.80,139.41,134.41,126.39, 125.98,123.67,122.04,120.36,119.46.44.18,30.42,29.45,27.25,25.34,18.93; HRMS(ESI)C 34 H 36 ClN5O2S2[M-Cl] + calcd=610.2305; found=610.2307.

[0189] Example 32 Compound 28

[0190] Compound 28 was synthesized by the method described in Example 5. The physicochemical properties of compound 28 are as follows:

[0191] 1) Yellow solid;

[0192] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0193] DMSO was used as solvent, and the peaks were assigned as follows: 1 HNMR(400MHz DMSO)δ:8.56(d,J=6.32Hz,2H,-Ph),8.17(dd,J=7.88,1.08Hz,1H,-Ph),7.93(d,J=7.08Hz,2H,-Ph),7.79(t,J=6.96Hz,1H ,-Ph),7.62-7.70(m,3H,-Ph),7.48(t,J=7.20Hz,1H,-Ph),7.36(t,J=7.32Hz,1H,-Ph),7.15(t,J=7.40Hz,1H,-Ph),5.50(s ,2H,-CH2-),4.87(t,J=7.16Hz,2H,-CH2-),4.42(t,J=6.68Hz,2H,-CH2-),3.50(t,J=6.76Hz,2H,-CH2-),3.26(t,J=7.24H z,4H,-CH2-),3.16(t,J=6.68Hz,2H,-CH2-),1.88-2.00(m,4H,-CH2-),1.74-1.86(m,4H,-CH2-),1.54-1.61(m,2H,-CH2-); 13 C NMR(100MHz DMSO)δ:162.94,162.88,160.53,146.80,145.35,143.70,139.41,134.40,126.88,126.39,125.98,125.10,123.67 .122.04,120.36,119.45,59.82,46.05,45.04,44.19,30.41,29.45,27.26,25.46,25.35,23.71,18.94; HRMS(ESI)C 34 H 36 ClN5O2S[M-Cl] + calcd=578.2584; found=578.2592.

[0194] Example 33 Compound 29

[0195] Compound 29 was synthesized by the method described in Example 5. The physicochemical properties of compound 29 are as follows:

[0196] 1) Yellow solid;

[0197] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0198] DMSO was used as solvent, and the peaks were assigned as follows: 1 HNMR(400MHz DMSO)δ:8.56(d,J=6.72Hz,2H,-Ph),8.16(dd,J=7.84,0.96Hz,1H,-Ph),7.92(d,J=6.80Hz,2H,-Ph),7.79(td,J=8.32,1.40Hz, 1H,-Ph),7.62-7.70(m,3H,-Ph),7.53-7.57(m,4H,-Ph)7.48(t,J=7.00Hz,2H,-Ph),7.35(t,J=7.32Hz,1H,-Ph),7.15(t,J=7.4 8Hz,1H,-Ph),5.16(s,2H,-CH2-),4.87(t,J=7.12Hz,2H,-CH2-),4.42(t,J=6.68Hz,2H,-CH2-),3.25(t,J=7.24Hz,2H,-CH2-), 3.23(s,3H,-CH3),3.16(t,J=6.72Hz,2H,-CH2-),1.86-1.93(m,2H,-CH2-),1.73-1.81(m,2H,-CH2-),1.53-1.61(m,2H,-CH2-); 13 C NMR(100MHz DMSO)δ:164.33,160.52,146.80,145.34,143.86,139.40,134.42,130.01,127.65,126.39,12 5.97,123.66,121.83,120.36,119.45,44.19,30.41,29.44,27.24,25.35,18.93; HRMS(ESI)C 37 H 36 ClN5O2S[M-Cl] + calcd=614.2584; found=614.2586.

[0199] Example 34 Compound 30

[0200] Compound 30 was synthesized by the method described in Example 5. The physicochemical properties of compound 30 are as follows:

[0201] 1) Yellow solid;

[0202] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400MHz) characteristics:

[0203] DMSO was used as solvent, and the peaks were assigned as follows: 1 HNMR(400MHz DMSO)δ:8.73(d,J=6.60Hz,1H,-Ph),8.59(d,J=6.92Hz,2H,-Ph),8.16(d,J=6.88Hz,1H,-NH-),7.90(d,J=7.04Hz,2H,-Ph),7.79(t,J =7.00Hz,1H,-Ph),7.62-7.70(m,3H,-Ph),7.48(t,J=7.16Hz,1H,-Ph),7.36(t,J=7.32Hz,1H,-Ph),7.15(t,J=7.36Hz,1H,-Ph),5.22( s,2H,-CH2-),4.87(t,J=7.28Hz,2H,-CH2-),4.42(t,J=6.68Hz,2H,-CH2-),3.97-4.05(m,1H,-CH-),3.25(t,J=7.24Hz,2H,-CH2-),3 .16(t,J=6.68Hz,2H,-CH2-),1.86-1.93(m,2H,-CH2-),1.75-1.84(m,4H,-CH2-),1.65-1.68(m,2H,-CH2-),1.42-1.59(m,6H,-CH2-); 13 C NMR(100MHz DMSO)δ:163.79,162.78,160.53,146.80,145.35,143.66,139.40,126.38,125.97,123.66,1 21.99,120.35,119.45,59.93,32.15,30.37,29.44,27.28,25.33,23.37,18.94; HRMS(ESI)C 35 H 38 ClN5O2S[M-Cl] + calcd=592.2741; found=592.2744.

[0204] Application Example 1: In vitro antibacterial activity assay

[0205] 1. Test bacteria:

[0206] Staphylococcus aureus (Staphylococcus aureus ATCC 29213); Escherichia coli (EscherichiacoliATCC25922); Methicillin-resistant Staphylococcus aureus (MRSA).

[0207] 2. Samples and reagents:

[0208] The samples are: evodiamine, vancomycin, meropenem and compounds 1-30 prepared in the examples.

[0209] 3. Test method:

[0210] According to the standards of the Clinical Laboratory Standards Institute (CLSI), the in vitro antibacterial activities of evodiamine, compounds 1-30 of the present invention and the clinical antibacterial drug vancomycin were tested using the broth microdilution method, and the minimum drug concentration that completely cleared the wells observed by naked eyes was taken as the MIC value.

[0211] Table 1. MIC values ​​of the pyridinium quaternary ammonium salt-containing evodia rutaecarpine derivatives 1-30 of the present invention against 10 MRSA clinical isolates and MRSA standard strain N315

[0212]

[0213]

[0214] Sa a .:S.aureusATCC 29213;M11-23 b :10 clinical MRSA strains; Ec c .:E.coli ATCC 25922; SI d :Selectivity coefficient (HC 50 / MICs of S.aureus); ND e : Not detected; Van f : Vancomycin; MEM g :Meropenem. The experiment was repeated at least 3 times.

[0215] It can be concluded from Table 1 that the quaternary pyridinium salt-containing evodiamine derivatives prepared by the present invention have significantly enhanced anti-Gram-positive bacteria activity compared to the substrate evodiamine. The synthesized compounds have no obvious antibacterial effect on E. coli. Compound 14 is the most significant compound in the 30 quaternary pyridinium salt-containing evodiamine derivatives against Gram-positive bacteria, with an MIC value between 0.5-2 μg / mL. Compound 14 has a relatively safe hemolytic activity (HC 50=282.63 / 277.231) and good membrane selectivity and safety (SI=565.26).

[0216] Application Example 2: Time-killing kinetics experiment of compound 14:

[0217] 1. Test bacteria:

[0218] S. aureus ATCC 29213; MRSA-11 (clinical isolate).

[0219] 2. Samples and reagents:

[0220] The samples were: vancomycin and compound 14 prepared in Example.

[0221] 3. Test method:

[0222] Monoclonal colonies of S. aureus ATCC 29213 and MRSA-11 were inoculated into 1 mL of MHB broth for overnight culture. 2 μL of bacterial solution was placed in 2 mL of MHB broth for expansion culture for 2.5 h (37 °C, 200 rpm). After the culture was completed, the bacterial concentration was determined to be 1×10 7 CFU / mL. Four groups of experiments were set up, namely 4×MIC / 8×MIC compound group, 8×MIC vancomycin group, and Control group. Compound 14 solution and vancomycin solution dissolved in DMSO were added to the cultured bacterial solution, and an equal amount of DMSO was added to the Control group. Then, the culture continued in a shaker, and 100 μL of bacterial solution was taken at 0, 0.5, 1, 2, 4, 6, and 8 h, respectively. After washing and resuspending, the samples were gradiently diluted in a 96-well plate. 10 μL of sample dilutions at different concentrations were aspirated and dropped onto the MHA solid culture dish, dried, and inverted in a 37°C incubator for 16-24 h. The number of colonies at different time points was read and a sterilization curve was drawn. Three parallel controls were set for each concentration. The experiment was repeated three times. The results are as follows Figure 1 As shown in A and B.

[0223] Figure 1A and B show that the bactericidal rate of compound 14 against MRSA-11 and S.aureus ATCC29213 is higher than that of vancomycin at the same 8×MIC concentration. For compound 14, the CFU / mL of MRSA-11 was reduced by more than 3 logarithms (killing 99.9% of bacteria) within 4h of 8×MIC and within 4h of 4×MIC. In addition, bacterial growth was also significantly inhibited under the action of 4×MIC concentration of compound 14. Compound 14 exhibits rapid bactericidal activity against S.aureusATCC29213, and can completely kill bacteria within 4 hours at 8×MIC. These results show that compound 14 has rapid bactericidal activity against Gram-positive bacteria MRSA-11 and S.aureusATCC29213.

[0224] Application Example 3: Drug resistance induction experiment of compound 14:

[0225] After determining the initial MIC values ​​of the target compound 14 and the positive control drug norfloxacin through drug sensitivity test, pick the monoclonal colonies of S. aureus ATCC 29213 and place them in two tubes of 1 mL MHB liquid culture medium, add the test compound and norfloxacin respectively, so that the final concentration is sub-inhibitory concentration, and culture for 12 hours. Use the inoculation loop to dip the bacterial solution, and use the three-zone streak method to inoculate the bacterial solution into the MHA solid culture medium containing 1 / 2MIC target compound and positive control drug and the solid culture medium without drug. After the monoclonal colonies appear, re-determine the MIC value and record it. The plate without drug is the first-generation MIC value, and the plate with drug is the second-generation MIC value. Repeat the above operation, and the sub-inhibitory concentration is adjusted according to the MIC value of each generation. A total of 20 generations were cultured. Results are shown in Figure 1 C.

[0226] from Figure 1 C The results showed that in the presence of sub-inhibitory concentration (1 / 2MIC) of compound 14, S. aureus ATCC29213 failed to produce resistant mutants after continuous passage within 20 days, indicating that the target was non-specific. After 4 passages, the MIC of norfloxacin began to rise and drug resistance appeared. After 18 passages, the MIC of norfloxacin increased by 128 times. This shows that the low drug resistance of compound 14 is beneficially affected by its rapid bactericidal activity.

[0227] Application Example 4: In vivo safety evaluation experiment - subcutaneous injection

[0228] 1. Reagents:

[0229] Compound 14 prepared in Example, 0.9% NaCl.

[0230] 2. Test animals

[0231] SPF grade KM female mice (purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd., weighing 20-22g, 7-8 weeks old).

[0232] 3. Test methods

[0233] After compound 14 was dissolved in DMSO, it was prepared into test compound solutions with concentrations of 80, 40, 20, 10, and 5 mg / kg with normal saline. 24 healthy female KM mice of the same size and weight were selected and randomly divided into six groups, including five experimental groups and one blank control group, with 4 mice in each group, and the backs were depilated. 60 μL of compound solutions of different concentrations were subcutaneously injected into the back of the mice in the experimental group, and 60 μL of normal saline was subcutaneously injected into the back of the mice in the blank control group. The survival status of the mice and whether there was an irritation reaction on the skin were observed. After 24 hours, blood was collected from the eyeballs of mice at the maximum injection concentration without death, frizzy hair and other irritation reactions, and blood routine, liver function, and kidney function tests were performed to observe whether there were significant changes compared with the blank control group.

[0234] The results showed that by subcutaneously injecting different concentrations of the compound into mice, when the concentration of compound 14 was ≤20 mg / kg, the mouse skin did not show any adverse reactions (such as redness, swelling, hardening, ulceration, etc.). Therefore, routine blood tests and blood biochemical index tests were performed on the mice at this dosage. Figure 2 The results showed that blood samples were subjected to routine blood tests and blood biochemistry analysis, and the test items included white blood cell count (WBC), red blood cell count (RBC), hemoglobin (HGB), hematocrit (HCT), mean corpuscular volume (MCV), platelet count (PLT), albumin (ALB), urea (UREA) and creatinine (CREA). The statistical results showed that compared with the blank control group (0.9% NaCl), after subcutaneous injection of compound 14 (20 mg / kg) into KM mice, there was no significant difference in the results, indicating that compound 14 has a certain in vivo safety.

[0235] Application Example 5: In vivo anti-MRSA infection activity experiment of compound 14 - subcutaneous injection

[0236] 1. Test bacteria:

[0237] MRSA-11 (clinical isolate)

[0238] 2. Samples and reagents:

[0239] The samples were: vancomycin, compound 14 prepared in Example, and 0.9% NaCl.

[0240] 3. Test animals:

[0241] SPF grade KM mice (purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd., weighing 20-22 g, 7-8 weeks old).

[0242] 4. Test method:

[0243] Thirty healthy female KM mice of the same size and weight were depilated from their backs and randomly divided into a blank group, a control group, a vancomycin group (5 mg / kg), a high-dose compound group (10 mg / kg), and a low-dose compound group (5 mg / kg), with 6 mice in each group. After the backs of the mice were disinfected, mice in the other groups except the blank group were subcutaneously injected with 60 μL of a 6×10 8 CFU / mL of MRSA-11 bacterial suspension. 2 hours later, 60 μL of different contents of compounds and vancomycin were injected in situ, and the control group was injected with normal saline. The survival and skin infection of each group of mice were recorded. 24 hours after administration, the mice in each group were killed by dislocation, and the skin of the infected area was taken and ground into a broken state using a tissue grinder. The grinding liquid of the skin tissue at the infected site was aspirated, and the number of colonies was read using the drop plate counting method. The results are as follows Figure 3 shown.

[0244] The results showed that the bacterial load in the skin tissue of mice in the Control group was about 6.9 log 10 CFU / g, while after treatment with low-dose 14 (5 mg / kg) and vancomycin (5 mg / kg), the number of MRSA cells in the mouse skin was significantly reduced (P<0.0001), and the bacterial load was reduced by 1.8 log 10 CFU / g, 1.4log 10 CFU / g, showing a moderate therapeutic effect. After 14 (10 mg / kg) treatment, the skin bacterial load decreased by about 5.4 log 10 CFU / g, the therapeutic effect is the most significant. Therefore, it is confirmed that the pyridinium quaternary ammonium salt-containing rutaecarpine derivative 14 has a good therapeutic effect on mouse skin abscesses infected with MRSA. The therapeutic effect is the same as that of the control drug vancomycin at the same dose. Compound 14 is expected to be developed into a new anti-MRSA drug.

[0245] Application Example 6: In vivo safety evaluation experiment - intraperitoneal injection

[0246] 1. Reagents:

[0247] Compound 14 prepared in Example, corn oil.

[0248] 2. Test animals

[0249] SPF BALB / c female mice (purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd., weighing 16-18 g, 7-8 weeks old).

[0250] 3. Test methods

[0251] After dissolving compound 14 with DMSO, corn oil was used to prepare the test compound solution with concentrations of 40, 20, 15, 10, and 5 mg / kg. 36 healthy female BALB / c mice of the same size and weight were selected and randomly divided into six groups, including five experimental groups and one blank control group, with 6 mice in each group. The mice in the experimental group were intraperitoneally injected with 100 μL of compound solutions of different concentrations, and the mice in the blank control group were subcutaneously injected with 100 μL of corn oil on the back. The mice were intraperitoneally injected once every 12 hours for two consecutive times. From the first day of administration, the mice were observed for seven consecutive days, and the survival status and survival rate of the mice were recorded. During this period, food and water were given normally. After determining the maximum tolerated dose, the drug was re-administered, and blood was collected from the eyeballs 24 hours later for routine blood tests and liver and kidney function tests to observe whether there were significant changes compared with the blank control group.

[0252] The results showed that by intraperitoneally injecting different concentrations of the compound into mice, when the concentration of compound 14 was ≤15 mg / kg, no adverse reactions (such as death, mental depression, hair tangling, etc.) occurred on the mouse skin. Therefore, routine blood tests and blood biochemical index tests were performed on the mice at this dosage. Figure 4 The results showed that blood samples were subjected to routine blood tests and blood biochemistry analysis, and the test items included white blood cell count (WBC), red blood cell count (RBC), hemoglobin (HGB), hematocrit (HCT), mean corpuscular volume (MCV), platelet count (PLT), albumin (ALB), urea (UREA) and creatinine (CREA). The statistical results showed that compared with the blank control group (corn oil), after intraperitoneal injection of compound 14 (15 mg / kg) into BALB / c mice, there was no significant difference in the results, indicating that compound 14 has a certain in vivo safety.

[0253] Application Example 7: In vivo anti-MRSA infection activity experiment of compound 14 - intraperitoneal injection

[0254] 1. Test bacteria:

[0255] MRSA-11 (clinical isolate)

[0256] 2. Samples and reagents:

[0257] The samples were: vancomycin, compound 14 prepared in Example, corn oil, and 0.9% NaCl.

[0258] 3. Test animals:

[0259] SPF BALB / c female mice (purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd., weighing 16-18 g, 7-8 weeks old).

[0260] 4. Test method:

[0261] 4.1 Determination of sublethal and lethal bacterial counts

[0262] SPF BALB / c mice were randomly divided into 7 groups, with 6 mice in each group. They were intraperitoneally injected with 100 μL of 1×10 8 , 5×10 8 , 1×10 9 , 5×10 9 , 1×10 10 , 1×10 11 The mice were intraperitoneally injected with saline containing CFU of bacteria, and the blank group was injected with 100 μL of saline. The mice were observed for seven consecutive days and given normal food and water. The status of the mice was recorded and the survival rate was calculated. After determining the minimum lethal dose and sublethal dose, the bacterial solution was re-injected. After 48 hours, the liver, spleen and kidney of each group of mice under the sublethal dose were homogenized, and the count was counted on the plate to calculate the bacterial load. Figure 5 shown.

[0263] 4.2 Organ bacterial burden and survival rate experiments

[0264] Thirty SPF BALB / c female mice of the same size and weight were randomly divided into a blank group, a control group, a vancomycin group (12 mg / kg), a low-dose compound group (6 mg / kg), and a high-dose compound group (12 mg / kg), with 6 mice in each group. The blank group received no medication or bacteria, while the other groups were injected with 100 μL of saline containing sublethal amounts of bacteria. 0.5 h later, the control group was intraperitoneally injected with 100 μL of corn oil, two groups were injected with 100 μL of compounds (6 mg / kg, 12 mg / kg), and one group was injected with the positive control drug vancomycin (12 mg / kg). The drugs were administered once every 12 h for two consecutive times. 24 h after the last administration, the mice were euthanized, and their livers, kidneys, and spleens were removed and ground into a broken state using a tissue grinder. The grinding fluid of the skin tissue at the infected site was aspirated, and the number of colonies was read using the drop plate counting method. The results are as follows: Figure 6 The liver, kidney and spleen of mice were also taken for tissue section observation.

[0265] Thirty SPF-grade BALB / c female mice of the same size and weight were randomly divided into a blank group, a control group, a vancomycin group (12 mg / kg), a low-dose compound group (6 mg / kg), and a high-dose compound group (12 mg / kg), with 6 mice in each group. The blank group received no medication or bacteria, while the other groups were injected with 100 μL of saline containing a lethal amount of bacteria. 0.5 h later, the control group was intraperitoneally injected with 100 μL of corn oil, two groups were injected with 100 μL of compounds (6 mg / kg, 12 mg / kg), and one group was injected with the positive control drug vancomycin (12 mg / kg). The drugs were administered once every 12 h for two consecutive times. The mice were observed for seven consecutive days, the survival status was recorded, and the mortality rate was calculated. The results are as follows Figure 7 shown.

Claims

1. A quaternary ammonium pyridinium salt-containing evodia rutaecarpine derivative, the structure of which is shown in the following formula (1): in, n=3,4 or 5, R is Among them, R 1 and R 2 Independently selected from H, phenyl, 4-6 membered cycloalkyl or C1-C5 alkyl, ring A is a 4-6 membered heterocycloalkyl containing one or two heteroatoms.

2. The quaternary pyridinium ammonium salt-containing evodia rutaecarpine derivative according to claim 1, characterized in that: The heteroatoms in the ring A, except the N atom connected to the parent nucleus, are absent or selected from one of N, O and S.

3. The quaternary pyridinium ammonium salt-containing evodia rutaecarpine derivative according to claim 1, characterized in that: The n, R is one of the following combinations: (1)n=3, (2)n=3, (3)n=3, (4)n=3, (5)n=3, (6)n=3, (7)n=3, (8)n=3, (9)n=3, (10)n=3, (11)n=4, (12)n=4, (13)n=4, (14)n=4, (15)n=4, (16)n=4, (17)n=4, (18)n=4, (19)n=4, (20)n=4, (21)n=5, (22)n=5, (23)n=5, (24)n=5, (25)n=5, (26)n=5, (27)n=5, (28)n=5, (29)n=5, (30)n=5, 4. The method for preparing the evodia rutaecarpine derivative containing a quaternary pyridinium ammonium salt according to any one of claims 1 to 3, characterized in that: The following steps are involved: (1) Using evodia rutaecarpine 1 as a substrate, it reacts with dibromoalkane 5 to obtain monobromoalkane-substituted intermediate 2; (2) intermediate 2 reacts with 4-mercaptopyridine to generate intermediate 3; (3) Chloroacetamide 6 reacts with intermediate 3 under the action of a catalyst to generate the quaternary pyridine ammonium salt-containing evodia rutaecarpine derivative 4, as shown in the following reaction formula: Wherein, n and R are the same as those described in any one of claims 1-3.

5. The method for preparing the evodia rutaecarpine derivative containing quaternary pyridinium ammonium salt according to claim 4, characterized in that: In step (1), the reaction molar ratio of the evodia rutaecarpine 1 to the dibromoalkane 5 is 1:(7.0-12.0), the catalyst is potassium carbonate, the reaction molar ratio of the evodia rutaecarpine 1 to potassium carbonate is 1:(6.5-7.0), the reaction temperature is 45±50°C, and the reaction solvent is acetone.

6. The method for preparing the evodia rutaecarpine derivative containing quaternary pyridinium ammonium salt according to claim 4, characterized in that: In step (2), the molar ratio of the reaction of the monobromoalkane-substituted intermediate 2 and 4-mercaptopyridine is 1:(1.0-1.5), the catalysts are potassium carbonate and potassium iodide, the molar ratio of the reaction of the intermediate 2 and potassium carbonate is 1:(1.0-1.7), and the molar ratio of the reaction with potassium iodide is 1:(0.01-0.03), the reaction temperature is 45±50°C, and the reaction solvent is acetone.

7. The method for preparing the evodia rutaecarpine derivative containing quaternary pyridinium ammonium salt according to claim 4, characterized in that: In step (3), the preparation method of chloroacetamide 6 comprises the following steps: chloroacetyl chloride and aliphatic amines / aromatic amines containing different substituents undergo substitution reaction under alkaline catalyst conditions to generate chloroacetamide 6: Preferably, the reaction molar ratio of the chloroacetyl chloride to the aliphatic amine / aromatic amine containing different substituents is (1.5-1.7):1, the alkaline catalyst is triethylamine, the reaction molar ratio of the aliphatic amine / aromatic amine containing different substituents to triethylamine is (1.0-1.2):1.5, the reaction temperature is 0±5°C, and the reaction solvent is anhydrous dichloromethane.

8. The method for preparing the evodia rutaecarpine derivative containing quaternary pyridinium ammonium salt according to claim 4, characterized in that: In step (3), the reaction molar ratio of the intermediate 3 to chloroacetamide 6 is 1:(2.0-2.5), nitrogen protection is adopted, the reaction temperature is 75±80°C, and the reaction solvent is anhydrous acetonitrile.

9. Use of the evodia rutaecarpine derivative containing a quaternary pyridinium ammonium salt according to any one of claims 1 to 3 in the preparation of antibacterial drugs.

10. The use according to claim 9, characterized in that: The antibacterial drug can inhibit Staphylococcus aureus or methicillin-resistant Staphylococcus aureus.

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