Evodiamine derivatives containing pyridine quaternary ammonium salt, preparation method and antibacterial application thereof

Abstract

The application discloses a series of evodiamine derivatives containing pyridine quaternary ammonium salt, a preparation method and antibacterial application thereof. The series of compounds are prepared by introducing a pyridine ring and then connecting with a chloroacetamide fragment based on a natural product evodiamine as a mother structure, so as to prepare a series of evodiamine derivatives containing pyridine quaternary ammonium salt, and the structural general formula is shown in the following formula (1). The series of compounds have strong in-vivo and in-vitro antibacterial activities on Staphylococcus aureus ATCC 29213 and clinically isolated Methicillin-resistant S. aureus (MRSA). The water solubility and antibacterial activity of the application are enhanced through further structure optimization, and the in-vivo and in-vitro toxicity, safe hemolytic activity and low drug resistance are low. Therefore, the evodiamine derivatives containing pyridine quaternary ammonium salt have the potential to be further developed as new antibacterial drugs.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of pharmaceutical chemistry, and specifically relates to pyridine quaternary ammonium salt-containing evonine derivatives, a preparation method thereof and antibacterial application of the evonine derivatives. BACKGROUND

[0002] Bacterial pathogens are the main cause of human diseases and deaths, and antibiotics are mainly used to treat infections caused by various bacteria. Due to the abuse of antibiotics, many pathogenic bacteria have evolved resistance to major classes 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, and induces various diseases after infection, from skin soft tissue infection to blood stream and internal organ infection, sepsis, pneumonia, meningitis, etc. At present, more than 60% of S. aureus isolates are resistant to methicillin (methicillin-resistant S. aureus / MRSA), MRSA has a gene encoding PBP2a in vivo, has low sensitivity to β-lactam antibiotics, and can synthesize cell wall under lethal β-lactam concentration. Peptidoglycan is the core component of the bacterial cell wall, and PBP2a enables the bacteria to maintain the biosynthesis of peptidoglycan and complete bacterial reproduction. (Ambade S S, Gupta V K, Bhole R P, et al. A review on five and six-membered heterocyclic compounds targeting the penicillin-binding protein 2 (PBP2A) of Methicillin-resistant Staphylococcus aureus (MRSA) [J]. Molecules, 2023, 28(20): 7008.) Antibiotic resistance has caused serious health and economic problems, and it is urgent to develop new antibacterial drugs with potential therapeutic effects on MRSA infection.

[0003] Quaternary Ammonium Salt is also called quaternary ammonium salt, which is easily soluble in water and has certain biological activity. Quaternary ammonium salt is an amphiphilic substance with good antibacterial performance, which can kill bacteria by electrostatic adsorption or insertion into the cell membrane / altering the permeability of the cell membrane, thereby reducing the probability of bacterial drug 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 fruits of Evodia rutaecarpa (Juss.) Benth. of Rutaceae. It has been widely studied for its treatment of gastrointestinal diseases, anti-tumor, hypertension and other related researches. However, there is little research on its antibacterial effect. Moreover, its water solubility and bioavailability are poor, making it difficult to be directly used as 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

[0004] The present application provides a pyridine quaternary ammonium salt-containing rutaecarpine derivative, a preparation method thereof and antibacterial applications thereof. The present application is based on the characteristics of quaternary ammonium salt as a cationic surfactant with antibacterial effect. The natural product rutaecarpine is used as the parent compound, and its structure is modified by introducing a pyridine ring and then connecting it with a chloroacetamide fragment to form a quaternary ammonium salt. The purpose is to obtain a pyridine quaternary ammonium salt-containing rutaecarpine derivative with high water solubility, high activity and low toxicity. The pyridine quaternary ammonium salt-containing rutaecarpine derivative has good antibacterial effect on Staphylococcus aureus (S. aureus) ATCC 29213, clinical isolated MRSA strains and MRSA standard strain N315, and solves the problems of high toxicity and poor water solubility.

[0005] Technical solutions: In order to achieve the above-mentioned purposes of the application, the technical solutions adopted by the present application are as follows:

[0006] The evodiamine derivative containing pyridine quaternary ammonium salt has the following formula (1):

[0007]

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

[0009] As a preferred solution, the heteroatoms in the ring A, in addition to the N atom connected to the mother nucleus, the rest of the heteroatoms are not present or selected from one of N, O, S.

[0010] As a preferred solution, the n, R is one of the following combinations:

[0011] (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 = CH3 (27) n = 5, R = CH3 (28) n = 5, R = CH3 (29) n = 5, R = CH3 (30) n = 5, R = CH3

[0012] The above specific selection represents compounds 1-30 in the following examples (each compound corresponds to the same number as described above).

[0013] The present application also provides a preparation method of the pyridine quaternary ammonium salt-containing evodiamine derivative, comprising the following steps:

[0014] (1) taking evodiamine 1 as a substrate, and reacting with dibromoalkane 5 to obtain a bromoalkyl-substituted intermediate 2;

[0015] (2) reacting the intermediate 2 with 4-mercaptopyridine to generate the intermediate 3;

[0016] (3) reacting the chloroacetamide 6 with the intermediate 3 under the action of a catalyst to generate the pyridine quaternary ammonium salt-containing evodiamine derivative 4, and the reaction formula is as shown below:

[0017]

[0018] wherein n and R are as described above.

[0019] As a specific embodiment, in step (1), the molar ratio of the evodiamine 1 to the dibromoalkane 5 is 1: (7.0-12.0), the catalyst is potassium carbonate, the molar ratio of the evodiamine 1 to the potassium carbonate is 1: (6.5-7.0), the reaction temperature is 45±55°C, and the reaction solvent is acetone.

[0020] As a specific embodiment, in step (2), the molar ratio of the intermediate 2 to 4-mercaptopyridine is 1: (1.0-1.5), the catalyst is potassium carbonate and potassium iodide, the molar ratio of the intermediate 2 to the potassium carbonate is 1: (1.0-1.7), and the molar ratio of the intermediate 2 to the potassium iodide is 1: (0.01-0.03), the reaction temperature is 45±50°C, and the reaction solvent is acetone.

[0021] As a specific embodiment, in step (3), the preparation method of the chloroacetamide 6 comprises the following steps: under the action of an alkaline catalyst, chloroacetyl chloride is subjected to a substitution reaction with a fatty amine / aromatic amine containing different substituents to generate the chloroacetamide 6:

[0022]

[0023] Preferably, the molar ratio of the chloroacetyl chloride to the aliphatic / aromatic amine with different substituents is (1.5-1.7) : 1, the basic catalyst is triethylamine, the molar ratio of the aliphatic / aromatic amine with different substituents to the triethylamine is (1.0-1.2) : 1.5, the reaction temperature is 0±5℃, and the reaction solvent is anhydrous dichloromethane.

[0024] More preferably, the molar ratio of the chloroacetyl chloride to the aliphatic / aromatic amine with different substituents is 1.5:1, the basic catalyst is triethylamine, the molar ratio of the aliphatic / aromatic amine with different substituents to the triethylamine is 1:1.5, the reaction temperature is 0℃, and the reaction solvent is anhydrous dichloromethane.

[0025] As a specific embodiment, in step (3), the molar ratio of the intermediate 3 to the chloroacetamide 6 is 1:(2.0-2.5), nitrogen protection is used, the reaction temperature is 75±80℃, and the reaction solvent is anhydrous acetonitrile.

[0026] Further, as a preferred embodiment:

[0027] In step (1), the molar ratio of the evodiamine to the dibromoalkane is 1:12, the catalyst is potassium carbonate, the molar ratio of the evodiamine to the potassium carbonate is 1:7, the reaction temperature is 50℃, and the reaction solvent is acetone.

[0028] In step (2), the molar ratio of the intermediate 2 to the 4-mercaptopyridine is 1:1, the catalyst is potassium carbonate and potassium iodide, the molar ratio of the intermediate 2 to the potassium carbonate is 1:1.5, the molar ratio to the potassium iodide is 1:0.02, the reaction temperature is 50℃, and the reaction solvent is acetone.

[0029] In step (3), the molar ratio of the intermediate 3 to the chloroacetamide is 1:2, nitrogen protection is used, the reaction temperature is 80℃, and the reaction solvent is anhydrous acetonitrile.

[0030] The application finally provides the use of the evodiamine derivative containing a pyridine quaternary ammonium salt 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 S. aureus ATCC 29213 and various clinical methicillin-resistant Staphylococcus aureus MRSA.

[0031] The compound of the present application connects natural product evonine with chloroacetamide fragment containing different substituents to generate quaternary ammonium salt, and designs and synthesizes a series of evonine derivatives containing quaternary ammonium pyridine salt by virtue of the characteristics of quaternary ammonium salt as cationic surfactant, which has antibacterial effect. The hydrophobic part of evonine-quaternary ammonium salt compound is beneficial to the insertion of the compound into the bacterial phospholipid bilayer membrane, and the positively charged cationic part is beneficial to the interaction with the negatively charged bacterial cell membrane, so as to cause the death of bacteria. The in-vitro antibacterial activity evaluation of all target compounds shows that all target compounds exhibit good in-vitro antibacterial activity on S.aureus ATCC 29213 and clinical isolated MRSA strains, and the minimum inhibitory concentration (MIC≤64 μg / mL).

[0032] In particular, the in-vitro and in-vivo anti-MRSA activity of the preferred compound 14 is most prominent, and the in-vivo antibacterial effect is equivalent to that of vancomycin at the same dose. In addition, the target compound has safe hemolytic activity, low toxicity, and plasma stability. Therefore, such compounds provide certain enlightenment for the research of new anti-MRSA drugs, and have wide application prospects.

[0033] Technical effects: The evonine derivatives containing quaternary ammonium pyridine salt prepared by the present application have good in-vitro and in-vivo bacteriostatic effect on S.aureus ATCC 29213 and various clinical MRSA strains, and have low biological toxicity, high yield, and are expected to be further developed into potential antibacterial drugs in clinic. BRIEF DESCRIPTION OF DRAWINGS

[0034] Figure 1 It is the dynamic bactericidal curve of compound 14.

[0035] Figure 2 It is the in-vivo blood routine and blood biochemical index of compound 14 (subcutaneous injection).

[0036] Figure 3 It is the change of bacterial load in mouse skin of compound 14 (subcutaneous injection).

[0037] Figure 4 It is the in-vivo blood routine and blood biochemical index of compound 14 (intraperitoneal injection).

[0038] Figure 5 It is the change of organ bacterial load of mice of compound 14 at lethal concentration and sublethal concentration (intraperitoneal injection).

[0039] Figure 6 It is the change of organ bacterial load of mice of compound 14 (intraperitoneal injection).

[0040] Figure 7 It is the survival rate of mice of compound 14 (intraperitoneal injection).

[0041] Figure 8 Compound 14 is 1 H-NMR spectrum.

[0042] Figure 9 Compound 14 is 13 C-NMR spectrum. DETAILED DESCRIPTION

[0043] The application is further described in detail by examples as follows.

[0044] Preparation of Example 1 Intermediate 2

[0045] In a 25 mL round-bottom flask, evodiamine (1 mmol) was dissolved in acetone as the reaction solvent, and dibromoalkane (12 mmol) and inorganic base potassium carbonate (7 mmol) were added. The mixture was stirred at 50°C under reflux in an oil bath for 72 h. After the reaction of evodiamine was completed, which was monitored by thin layer chromatography (TLC), the acetone in the reaction solution was concentrated under reduced pressure, and the mixture was extracted with dichloromethane and dried over anhydrous sodium sulfate. The concentrated solution was purified by silica gel column chromatography using petroleum ether: ethyl acetate = 5:1 as the developing agent to obtain white solid intermediate 2.

[0046] Preparation of Example 2 Intermediate 3

[0047] After intermediate 2 (1 mmol) was dissolved in an appropriate amount of acetone, 4-mercaptopyridine (1 mmol), a catalyst inorganic base potassium carbonate (1.5 mmol), and a catalyst potassium iodide (0.02 mmol) were added. The mixture was stirred at 50°C under reflux in an oil bath for 12 h. After the reaction of intermediate 2 was completed, which was monitored by thin layer chromatography (TLC), the acetone in the reaction solution was concentrated under reduced pressure, and the mixture was extracted with dichloromethane and dried over anhydrous sodium sulfate. The concentrated solution was purified by silica gel column chromatography using dichloromethane:methanol:acetone = 35:1:1 as the developing agent to obtain yellow solid intermediate 3.

[0048] Preparation of Example 3 Chloroacetamide

[0049] In a 25 mL round-bottom flask, an aliphatic amine / aromatic amine containing different substituents (1 mmol) was added, and an appropriate amount of dichloromethane was added as the reaction solvent. The mixture was placed in an ice bath (0°C) and chloroacetyl chloride (1.5 mmol) was slowly added dropwise over 10 min. After stirring for 2-3 min, triethylamine (1.5 mmol) was quickly added, and the reaction was carried out under nitrogen protection for 5 h. After the reaction was completed, which was monitored by thin layer chromatography (TLC), the mixture was extracted with dichloromethane and dried over anhydrous sodium sulfate. The concentrated solution was purified by silica gel column chromatography using dichloromethane:methanol = 10:1 as the developing agent to obtain brown oil chloroacetamide containing different substituents.

[0050] Preparation of Example 4 intermediate 4

[0051] Dissolve intermediate 3 (1 mmol) in an appropriate amount of anhydrous acetonitrile as the reaction solvent, add chloroacetamide (2 mmol), and stir under reflux at 80°C in an oil bath under nitrogen protection for 12 h. After monitoring the completion of the reaction by thin layer chromatography (TLC), purify by silica gel column chromatography with dichloromethane:methanol = 10:1 to obtain the evodiamine quaternary ammonium salt compound as a light yellow solid.

[0052] Example 5 Compound 1

[0053] Dissolve intermediate 3 (1 mmol) in an appropriate amount of anhydrous acetonitrile as the reaction solvent, add chloroacetamide (2 mmol), and stir under reflux at 80°C in an oil bath under nitrogen protection for 12 h. After monitoring the completion of the reaction by thin layer chromatography (TLC), purify by silica gel column chromatography with dichloromethane:methanol = 10:1 to obtain the evodiamine quaternary ammonium salt compound as a light yellow solid.

[0054] The physicochemical properties of Compound 1 are as follows:

[0055] 1), white solid;

[0056] 2), the nuclear magnetic resonance spectrum of the compound (H / 1 H / 13 C NMR, 400 MHz) characteristics:

[0057] In CDCl3 as the solvent, each peak is assigned as follows: 1H NMR (400 MHz CDCl3) δ: 8.73 (d, J = 6.48 Hz, 2H, -Ph), 8.31 (dd, J = 7.92, 1.08 Hz, 1H, -Ph), 7.75 (t, J = 6.96 Hz, 1H, -Ph), 7.66 (d, J = 5.76 Hz, 2H, -Ph), 7.42-7.47 (m, 4H, -Ph), 7.39 (d, J = 7.68 Hz, 1H, -Ph), 7.21 (t, J = 7.16 Hz, 1H, -Ph), 6.04 (s, 2H, -CH2-), 5.06 (t, J = 6.72 Hz, 2H, -CH2-), 4.54 (t, J = 6.72 Hz, 2H, -CH2-), 3.46 (t, J = 7.64 Hz, 2H, -CH2-), 3.30 (t, J = 7.6 Hz, 2H, -S-CH2-), 3.18-3.24 (m, 4H, -CH2-), 2.43-2.50 (m, 2H, -CH2-), 1.61-1.69 (m, 2H, -CH2-), 1.47-1.55 (m, 2H, -CH2-), 1.37-1.43 (m, 2H, -CH2-), 1.24-1.31 (m, 2H, -CH2-), 0.97 (t, J = 7.28 Hz, 3H, -CH3), 0.90 (t, J = 7.28 Hz, 3H, -CH3); HRMS (ESI) C 36 H 42 ClN5O2S[M-Cl] + calcd = 608.3054; found = 608.3061.

[0058] Example 6 Compound 2

[0059] Compound 2 was synthesized by the method described in Example 5, and the physical and chemical properties of Compound 2 were as follows:

[0060] 1) white solid;

[0061] 2) the nuclear magnetic resonance spectrum (H / 1 H / 13 C NMR, 400 MHz) characteristics of the compound:

[0062] In CDCl3as solvent, each peak is assigned as: 1HNMR (400 MHz CDCl3) δ: 8.74 (d, J = 5.92 Hz, 2H, -Ph), 8.31 (d, J = 7.32 Hz, 1H, -Ph), 7.75 (t, J = 7.08 Hz, 1H, -Ph), 7.66 (dd, J = 7.84, 3.2 Hz, 2H, -Ph), 7.41-7.45 (m, 4H, -Ph), 7.39 (d, J = 8.08 Hz, 1H, -Ph), 7.20 (t, J = 7.2 Hz, 1H, -Ph), 6.06 (s, 2H, -CH2-), 5.06 (t, J = 6.64 Hz, 2H, -CH2-), 4.53 (t, J = 6.72 Hz, 2H, -CH2-), 3.43 (t, J = 7.52 Hz, 2H, -CH2-), 3.27 (t, J = 7.68 Hz, 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 = 7.28 Hz, 3H, -CH3), 0.87 (t, J = 7.32 Hz, 3H, -CH3); HRMS (ESI) C 34 H 38 ClN5O2S[M-Cl] + calcd = 580.2741; found = 580.2745.

[0063] Example 7 Compound 3

[0064] Compound 3 was synthesized using the method described in Example 5, and the physical and chemical properties of Compound 3 are as follows:

[0065] 1) white solid;

[0066] 2) the nuclear magnetic resonance spectrum (H 1 H / 13 C NMR, 400 MHz) characteristics of the compound are as follows:

[0067] In CDCl3 as solvent, wherein each peak is assigned as: 1HNMR (400 MHz CDCl3) δ: 9.28 (t, J = 5.44 Hz, 1H, -Ph), 8.81 (d, J = 6.76 Hz, 2H, -Ph), 8.32 (dd, J = 7.92, 1.16 Hz, 1H, -NH-), 7.76 (t, J = 6.88 Hz, 1H, -Ph), 7.68 (dd, J = 12.48, 7.96 Hz, 2H, -Ph), 7.43-7.48 (m, 4H, -Ph), 7.40 (td, J = 7.68, 0.84 Hz, 1H, -Ph), 7.22 (td, J = 8.8, 7.8 Hz, 1H, -Ph), 5.59 (s, 2H, -CH2-), 5.09 (t, J = 6.6 Hz, 2H, -CH2-), 4.54 (t, J = 6.76 Hz, 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 = 7.32 Hz, 3H, -CH3); HRMS (ESI) C 32 H 34 ClN5O2S[M-Cl] + calcd = 552.2428; found = 552.2435.

[0068] Example 8 Compound 4

[0069] Compound 4 was synthesized using the method described in Example 5. The physicochemical properties of compound 4 are as follows:

[0070] 1) yellow solid;

[0071] 2) the nuclear magnetic resonance spectrum (H NMR, 400 MHz) of the compound is characterized by: 1 H / 13 C NMR, 400 MHz) is characterized by:

[0072] in DMSO, in which each peak is assigned as: 1HNMR (400 MHz DMSO) δ: 8.63 (t, J = 5.52 Hz, 1H, -Ph), 8.59 (d, J = 7.04 Hz, 2H, -Ph), 8.17 (dd, J = 7.88, 1.08 Hz, 1H, -NH-), 7.98 (d, J = 7.08 Hz, 2H, -Ph), 7.82 (td, J = 8.28, 1.44 Hz, 1H, -Ph), 7.69-7.33 (m, 3H, -Ph), 7.49 (t, J = 7.84 Hz, 1H, -Ph), 7.38 (t, J = 7.4 Hz, 1H, -Ph), 7.17 (t, J = 7.56 Hz, 1H, -Ph), 5.23 (s, 2H, -CH2-), 5.02 (t, J = 6.92 Hz, 2H, -CH2-), 4.43 (t, J = 6.72 Hz, 2H, -CH2-), 3.45 (t, J = 6.96 Hz, 2H, -CH2-), 3.17 (t, J = 6.64 Hz, 2H, -CH2-), 3.10 (t, J = 5.84 Hz, 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 = 6.80 Hz, 3H, -CH3); HRMS (ESI) C 33 H 36 ClN5O2S[M-Cl] + calcd = 566.2584; found = 566.2589.

[0073] Example 9 Compound 5

[0074] Compound 5 was synthesized using the method described in Example 5, and the physical and chemical properties of compound 5 are as follows:

[0075] 1) white solid;

[0076] 2) the nuclear magnetic resonance spectrum of the compound (H NMR, 400 MHz) is characterized by: 1 H / 13 C NMR, 400 MHz) is characterized by:

[0077] in DMSO, wherein each peak is assigned as: 1HNMR (400 MHz DMSO) δ: 8.67 (t, J = 5.40 Hz, 1H, -Ph), 8.60 (d, J = 7.08 Hz, 2H, -Ph), 8.16 (dd, J = 7.96, 1.2 Hz, 1H, -NH-), 7.98 (d, J = 7.16 Hz, 2H, -Ph), 7.82 (td, J = 6.84, 1.48 Hz, 1H, -Ph), 7.69-7.73 (m, 3H, -Ph), 7.49 (td, J = 7.04, 0.96 Hz, 1H, -Ph), 7.38 (td, J = 7.32, 0.8 Hz, 1H, -Ph), 7.17 (t, J = 7.64 Hz, 1H, -Ph), 5.24 (s, 2H, -CH2-), 5.02 (t, J = 6.96 Hz, 2H, -CH2-), 4.43 (t, J = 6.72 Hz, 2H, -CH2-), 3.46 (t, J = 6.96 Hz, 2H, -CH2-), 3.17 (t, J = 6.72 Hz, 2H, -CH2-), 3.09 (q, J = 6.84 Hz, 2H, -CH2-), 2.27-2.32 (m, 2H, -CH2-), 1.42-1.51 (m, 2H, -CH2-), 0.88 (t, J = 7.36 Hz, 3H, -CH3); HRMS (ESI) C 31 H 32 ClN5O2S[M-Cl] + calcd = 538.2271 ; found = 538.2278.

[0078] Example 10 Compound 6

[0079] Compound 6 was synthesized using the method described in Example 5. The physical and chemical properties of compound 6 are as follows:

[0080] 1) white solid;

[0081] 2) the nuclear magnetic resonance spectrum of the compound (H NMR, 400 MHz) is characterized by: 1 H / 13 C NMR, 400 MHz) is characterized by:

[0082] in DMSO, in which each peak is assigned as: 1HNMR (400 MHz DMSO) δ: 8.57 (d, J = 7.08 Hz, 2H, -Ph), 8.16 (dd, J = 7.92, 1.2 Hz, 1H, -Ph), 8.01 (d, J = 7.12 Hz, 2H, -Ph), 7.82 (td, J = 8.32, 1.44 Hz, 1H, -Ph), 7.72 (t, J = 5.12 Hz, 3H, -Ph), 7.50 (td, J = 6.24, 0.8 Hz, 1H, -Ph), 7.38 (t, J = 7.12 Hz, 1H, -Ph), 7.18 (t, J = 7.68 Hz, 1H, -Ph), 5.60 (s, 2H, -CH2-), 5.03 (t, J = 6.84 Hz, 2H, -CH2-), 4.43 (t, J = 6.72 Hz, 2H, -CH2-), 3.70 (t, J = 4.28 Hz, 2H, -CH2-), 3.61 (t, J = 4.32 Hz, 2H, -CH2-), 3.44-3.48 (m, 6H, -O-CH2-, -N-CH2-), 3.17 (t, J = 6.72 Hz, 2H, -CH2-), 2.28-2.35 (m, 2H, -CH2-), 13 C NMR (100 MHz 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.

[0083] Example 11 Compound 7

[0084] Compound 7 was synthesized using the method described in Example 5. The physical and chemical properties of compound 7 are as follows:

[0085] 1) yellow solid;

[0086] 2) the nuclear magnetic resonance spectrum (H / 1 H / 13 C NMR, 400 MHz) characteristics of the compound are as follows:

[0087] in DMSO, in which each peak is 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.

[0088] Example 12 Compound 8

[0089] Compound 8 was synthesized using the method described in Example 5. The physicochemical properties of compound 8 are as follows:

[0090] 1) Yellow solid;

[0091] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400 MHz) characteristics:

[0092] With DMSO as solvent, the peaks are attributed to: 1HNMR (400 MHz DMSO) δ: 8.57 (d, J = 6.96 Hz, 2H, -Ph), 8.17 (d, J = 7.20 Hz, 1H, -Ph), 8.01 (d, J = 7.00 Hz, 2H, -Ph), 7.83 (t, J = 7.04 Hz, 1H, -Ph), 7.72 (td, J = 4.64, 3.96 Hz, 3H, -Ph), 7.50 (t, J = 7.52 Hz, 1H, -Ph), 7.38 (t, J = 7.32 Hz, 1H, -Ph), 7.18 (t, J = 7.60 Hz, 1H, -Ph), 5.48 (s, 2H, -CH2-), 5.02 (t, J = 6.76 Hz, 2H, -CH2-), 4.43 (t, J = 6.68 Hz, 2H, -CH2-), 3.43 - 3.51 (m, 6H, -CH2-), 3.17 (t, J = 6.68 Hz, 2H, -CH2-), 2.27 - 2.34 (m, 2H, -CH2-), 1.93 - 2.00 (m, 2H, -CH2-), 1.79 - 1.86 (m, 2H, -CH2-); HRMS (ESI) C 32 H 32 ClN5O2S[M-Cl] + calcd = 550.2271 ; found = 550.2280.

[0093] Example 13 Compound 9

[0094] Compound 9 was synthesized using the method described in Example 5. The physical and chemical properties of compound 9 are as follows:

[0095] 1) yellow solid;

[0096] 2) the nuclear magnetic resonance spectrum (H NMR, 400 MHz) of the compound is characterized by: 1 H / 13 C NMR, 400 MHz) is as follows:

[0097] in DMSO, wherein each peak is assigned as: 1HNMR (400 MHz DMSO) δ: 8.58 (d, J = 6.76 Hz, 2H, -Ph), 8.16 (dd, J = 7.88, 1.08 Hz, 1H, -Ph), 8.00 (d, J = 6.92 Hz, 2H, -Ph), 7.82 (td, J = 8.32, 1.44 Hz, 1H, -Ph), 7.71 (t, J = 6.64 Hz, 3H, -Ph), 7.56-7.58 (m, 4H, -Ph), 7.49 (td, J = 8.00, 0.96 Hz, 2H, -Ph), 7.38 (t, J = 7.12 Hz, 1H, -Ph), 7.17 (t, J = 7.60 Hz, 1H, -Ph), 5.16 (s, 2H, -CH2-), 5.02 (t, J = 6.92 Hz, 2H, -CH2-), 4.42 (t, J = 6.72 Hz, 2H, -CH2-), 3.46 (t, J = 7.00 Hz, 2H, -CH2-), 3.23 (s, 3H, -CH3), 3.17 (t, J = 6.72 Hz, 2H, -CH2-), 2.27-2.34 (m, 2H, -CH2-); HRMS (ESI) C 35 H 32 ClN5O2S[M-Cl] + calcd = 586.2271 ; found = 586.2279.

[0098] Example 14 Compound 10

[0099] Compound 10 was synthesized using the method described in Example 5. The physical and chemical properties of compound 10 are as follows:

[0100] 1) yellow solid;

[0101] 2) the nuclear magnetic resonance spectrum (H NMR, 400 MHz) of the compound is characterized by: 1 H / 13 C NMR, 400 MHz) is characterized by:

[0102] in DMSO, wherein each peak is assigned as: 1HNMR (400 MHz DMSO) δ: 8.62 (d, J = 7.20 Hz, 1H, -Ph), 8.58 (d, J = 7.08 Hz, 2H, -Ph), 8.17 (dd, J = 7.96, 1.20 Hz, 1H, -NH-), 7.97 (d, J = 7.12 Hz, 2H, -Ph), 7.82 (t, J = 6.84 Hz, 1H, -Ph), 7.69-7.73 (m, 3H, -Ph), 7.49 (t, J = 8.00 Hz, 1H, -Ph), 7.38 (t, J = 7.24 Hz, 1H, -Ph), 7.17 (t, J = 7.40 Hz, 1H, -Ph), 5.16 (s, 2H, -CH2-), 5.02 (t, J = 6.64 Hz, 2H, -CH2-), 4.43 (t, J = 6.80 Hz, 2H, -CH2-), 3.96-4.05 (m, 1H, -CH-), 3.45 (t, J = 6.84 Hz, 2H, -CH2-), 3.17 (t, J = 6.76 Hz, 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 (100 MHz 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.

[0103] Example 15 Compound 11

[0104] Compound 11 was synthesized using the method described in Example 5, and the physical and chemical properties of compound 11 are as follows:

[0105] 1) yellow solid;

[0106] 2) the nuclear magnetic resonance spectrum (H / 1 H / 13 C NMR, 400 MHz) characteristics of this compound are:

[0107] in DMSO, in which each peak is assigned as: 1HNMR (400 MHz DMSO) δ: 8.60 (d, J = 6.80 Hz, 2H, -Ph), 8.16 (d, J = 7.96 Hz, 1H, -Ph), 7.93 (d, J = 7.00 Hz, 2H, -Ph), 7.78 (t, J = 6.96 Hz, 1H, -Ph), 7.69 (t, J = 8.72 Hz, 2H, -Ph), 7.62 (d, J = 8.08 Hz, 1H, -Ph), 7.47 (t, J = 7.12 Hz, 1H, -Ph), 7.37 (t, J = 7.28 Hz, 1H, -Ph), 7.16 (t, J = 7.24 Hz, 1H, -Ph), 5.61 (s, 2H, -CH2-), 4.92 (t, J = 7.00 Hz, 2H, -CH2-), 4.43 (t, J = 6.72 Hz, 2H, -CH2-), 3.27-3.37 (m, 6H, -CH2-), 3.17 (t, J = 6.60 Hz, 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 = 7.24 Hz, 3H, -CH3), 0.87 (t, J = 7.28 Hz, 3H, -CH3); HRMS (ESI) C 37 H 44 ClN5O2S[M-Cl] + calcd = 622.3210; found = 622.3214.

[0108] Example 16 Compound 12

[0109] Compound 12 was synthesized using the method described in Example 5. The physical and chemical properties of compound 12 are as follows:

[0110] 1) yellow solid;

[0111] 2) the nuclear magnetic resonance spectrum (H NMR, 400 MHz) of the compound is characterized by: 1 H / 13 C NMR, 400 MHz) is as follows:

[0112] in DMSO, wherein each peak is assigned as: 1HNMR (400 MHz DMSO) δ: 8.60 (d, J = 7.08 Hz, 2H, -Ph), 8.17 (dd, J = 7.96, 1.16 Hz, 1H, -Ph), 7.93 (d, J = 7.12 Hz, 2H, -Ph), 7.78 (t, J = 6.88 Hz, 1H, -Ph), 7.69 (t, J = 8.76 Hz, 2H, -Ph), 7.62 (d, J = 7.88 Hz, 1H, -Ph), 7.47 (t, J = 7.92 Hz, 1H, -Ph), 7.37 (t, J = 7.24 Hz, 1H, -Ph), 7.16 (t, J = 7.44 Hz, 1H, -Ph), 5.60 (s, 2H, -CH2-), 4.92 (t, J = 7.12 Hz, 2H, -N-CH2-), 4.43 (t, J = 6.68 Hz, 2H, -N-CH2-), 3.24-3.35 (m, 6H, -N-CH2-, -CH2-), 3.17 (t, J = 6.64 Hz, 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 = 7.28 Hz, 3H, -CH3), 0.83 (t, J = 7.36 Hz, 3H, -CH3), 13 C NMR (100 MHz 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.

[0113] Example 17 Compound 13

[0114] Compound 13 was synthesized using the method described in Example 5. The physicochemical properties of compound 13 are as follows:

[0115] 1) yellow solid;

[0116] 2) the nuclear magnetic resonance spectrum of the compound 1 H / 13C NMR, 400 MHz) characteristics:

[0117] In DMSO as solvent, where each peak is assigned: 1 HNMR (400 MHz DMSO) δ: 8.71 (t, J = 5.40 Hz, 1H, -Ph), 8.60 (d, J = 7.12 Hz, 2H, -Ph), 8.16 (dd, J = 7.96, 1.28 Hz, 1H, -NH-), 7.91 (d, J = 7.16 Hz, 2H, -Ph), 7.78 (t, J = 6.88 Hz, 1H, -Ph), 7.68 (t, J = 8.92 Hz, 2H, -Ph), 7.62 (d, J = 7.84 Hz, 1H, -Ph), 7.47 (t, J = 7.04 Hz, 1H, -Ph), 7.37 (t, J = 7.20 Hz, 1H, -Ph), 7.16 (t, J = 7.32 Hz, 1H, -Ph), 5.27 (s, 2H, -CH2-), 4.91 (t, J = 7.08 Hz, 2H, -CH2-), 4.42 (t, J = 6.72 Hz, 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 = 7.28 Hz, 3H, -CH3), 13 C NMR (100 MHz 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.

[0118] Compound 14 of Example 18

[0119] Compound 14 was synthesized using the method described in Example 5, and the physicochemical properties of compound 14 are as follows:

[0120] 1) yellow solid;

[0121] 2), the nuclear magnetic resonance spectrum of the compound 1 H / 13 C NMR, 400MHz) characteristics:

[0122] In DMSO as solvent, each peak is assigned as follows: 1 H NMR (400MHz DMSO) δ: 8.71 (t, J = 5.44 Hz, 1H, -Ph), 8.60 (d, J = 7.16 Hz, 2H, -Ph), 8.16 (dd, J = 7.96, 1.24 Hz, 1H, -NH-), 7.91 (d, J = 7.16 Hz, 2H, -Ph), 7.78 (td, J = 8.36, 1.48 Hz, 1H, -Ph), 7.68 (t, J = 9.00 Hz, 2H, -Ph), 7.62 (d, J = 7.88 Hz, 1H, -Ph), 7.47 (td, J = 8.00, 0.96 Hz, 1H, -Ph), 7.37 (t, J = 7.24 Hz, 1H, -Ph), 7.16 (t, J = 7.56 Hz, 1H, -Ph), 5.26 (s, 2H, -CH2-), 4.91 (t, J = 7.08 Hz, 2H, -CH2-), 4.42 (t, J = 6.68 Hz, 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 = 6.76 Hz, 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, 122.04, 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.

[0123] Compound 15 of Example 19

[0124] Compound 15 was synthesized by the method described in Example 5, and the physicochemical properties of compound 15 were as follows:

[0125] 1) yellow solid;

[0126] 2) the nuclear magnetic resonance spectrum of the compound 1 H / 13 C NMR, 400MHz) characteristics:

[0127] In DMSO as solvent, each peak is assigned as follows: 1 HNMR (400MHz DMSO) δ: 8.73 (t, J = 4.96 Hz, 1H, -Ph), 8.61 (d, J = 7.04 Hz, 2H, -Ph), 8.16 (dd, J = 7.88, 1.16 Hz, 1H, -NH-), 7.91 (d, J = 7.12 Hz, 2H, -Ph), 7.78 (t, J = 6.92 Hz, 1H, -Ph), 7.69 (t, J = 8.88 Hz, 2H, -Ph), 7.62 (d, J = 7.96 Hz, 1H, -Ph), 7.47 (t, J = 7.08 Hz, 1H, -Ph), 7.37 (t, J = 7.24 Hz, 1H, -Ph), 7.16 (t, J = 7.28 Hz, 1H, -Ph), 5.27 (s, 2H, -CH2-), 4.91 (t, J = 7.00 Hz, 2H, -CH2-), 4.42 (t, J = 6.76 Hz, 2H, -CH2-), 3.35-3.36 (m, 2H, -CH2-), 3.17 (t, J = 6.68 Hz, 2H, -CH2-), 3.10 (q, J = 6.76 Hz, 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 = 7.36 Hz, 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, 125.98, 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.

[0128] Example 20 Compound 16

[0129] Compound 16 was synthesized using the method described in Example 5. The physicochemical properties of compound 16 are as follows:

[0130] 1) Yellow solid;

[0131] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400 MHz) characteristics:

[0132] With DMSO as solvent, the peaks are attributed to: 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.

[0133] Example 21 Compound 17

[0134] Compound 17 was synthesized using the method described in Example 5. The physicochemical properties of compound 17 are as follows:

[0135] 1) Yellow solid;

[0136] 2) The NMR spectrum of the compound ( 1 H / 13C NMR, 400 MHz) characteristics:

[0137] In DMSO as solvent, each peak is assigned as follows: 1 HNMR (400 MHz DMSO) δ: 8.57 (d, J = 6.68 Hz, 2H, -Ph), 8.16 (dd, J = 7.84, 1.04 Hz, 1H, -Ph), 7.94 (d, J = 7.04 Hz, 2H, -Ph), 7.78 (t, J = 6.88 Hz, 1H, -Ph), 7.69 (t, J = 8.12 Hz, 2H, -Ph), 7.62 (d, J = 7.96 Hz, 1H, -Ph), 7.48 (t, J = 7.84 Hz, 1H, -Ph), 7.37 (t, J = 7.68 Hz, 1H, -Ph), 7.16 (t, J = 7.48 Hz, 1H, -Ph), 5.61 (s, 2H, -CH2-), 4.92 (t, J = 7.04 Hz, 2H, -CH2-), 4.43 (t, J = 6.64 Hz, 2H, -CH2-), 3.72-3.75 (m, 4H, -CH2-), 3.34-3.37 (m, 2H, -CH2-), 3.17 (t, J = 6.68 Hz, 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 (100 MHz DMSO) δ: 163.53, 162.95, 160.51, 146.77, 145.30, 143.78, 139.44, 134.40, 126.37, 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.

[0138] Example 22 Compound 18

[0139] Compound 18 was synthesized using the method described in Example 5, and the physicochemical properties of compound 18 are as follows:

[0140] 1) yellow solid;

[0141] 2) the nuclear magnetic resonance spectrum of the compound 1 H / 13C NMR, 400 MHz) features:

[0142] In DMSO as solvent, each peak is assigned as follows: 1 HNMR (400 MHz DMSO) δ: 8.57 (d, J = 6.84 Hz, 2H, -Ph), 8.16 (dd, J = 7.92, 1.12 Hz, 1H, -Ph), 7.93 (d, J = 7.12 Hz, 2H, -Ph), 7.78 (t, J = 6.88 Hz, 1H, -Ph), 7.69 (t, J = 8.92 Hz, 2H, -Ph), 7.62 (d, J = 8.04 Hz, 1H, -Ph), 7.48 (t, J = 7.88 Hz, 1H, -Ph), 7.37 (t, J = 7.92 Hz, 1H, -Ph), 7.16 (t, J = 7.56 Hz, 1H, -Ph), 5.51 (s, 2H, -CH2-), 4.92 (t, J = 6.96 Hz, 2H, -CH2-), 4.43 (t, J = 6.72 Hz, 2H, -CH2-), 3.51 (t, J = 6.72 Hz, 2H, -CH2-), 3.36-3.37 (m, 4H, -CH2-), 3.17 (t, J = 6.68 Hz, 2H, -CH2-), 1.94-2.05 (m, 4H, -CH2-), 1.81-1.86 (m, 4H, -CH2-), 13 C NMR (100 MHz DMSO) δ: 162.89, 162.84, 160.51, 146.78, 145.30, 143.70, 139.45, 134.40, 126.37, 125.97, 123.68, 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.

[0143] Compound 19 of Example 23

[0144] Compound 19 was synthesized using the method described in Example 5, and the physical and chemical properties of compound 19 are as follows:

[0145] 1) yellow solid;

[0146] 2) the nuclear magnetic resonance spectrum of the compound 1 H / 13 C NMR, 400 MHz) features:

[0147] With DMSO as solvent, the peaks are attributed to: 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.

[0148] Example 24 Compound 20

[0149] Compound 20 was synthesized using the method described in Example 5. The physicochemical properties of compound 20 are as follows:

[0150] 1) Yellow solid;

[0151] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400 MHz) characteristics:

[0152] With DMSO as solvent, the peaks are attributed to: 1HNMR (400 MHz DMSO) δ: 8.69 (d, J = 6.84 Hz, 1H, -Ph), 8.59 (d, J = 7.00 Hz, 2H, -Ph), 8.17 (dd, J = 7.92, 1.12 Hz, 1H, -NH-), 7.90 (d, J = 7.04 Hz, 2H, -Ph), 7.78 (t, J = 6.96 Hz, 1H, -Ph), 7.69 (t, J = 8.00 Hz, 2H, -Ph), 7.62 (d, J = 8.04 Hz, 1H, -Ph), 7.48 (t, J = 7.12 Hz, 1H, -Ph), 7.37 (t, J = 7.32 Hz, 1H, -Ph), 7.16 (t, J = 7.32 Hz, 1H, -Ph), 5.20 (s, 2H, -CH2-), 4.91 (t, J = 7.08 Hz, 2H, -CH2-), 4.42 (t, J = 6.76 Hz, 2H, -CH2-), 3.98-4.06 (m, 1H, -CH-), 3.33-3.35 (m, 2H, -CH2-), 3.17 (t, J = 6.68 Hz, 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 (100 MHz DMSO) δ: 163.79, 162.69, 160.51, 146.78, 145.30, 143.64, 139.44, 134.42, 126.37, 125.96, 123.67, 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.

[0153] Example 25 Compound 21

[0154] Compound 21 was synthesized using the method described in Example 5, and the physical and chemical properties of compound 21 are as follows:

[0155] 1) yellow solid;

[0156] 2) the nuclear magnetic resonance spectrum of this compound (H 1 H / 13 C NMR, 400 MHz) characteristics:

[0157] With DMSO as solvent, the peaks are attributed to: 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.

[0158] Example 26 Compound 22

[0159] Compound 22 was synthesized using the method described in Example 5. The physicochemical properties of compound 22 are as follows:

[0160] 1) Yellow solid;

[0161] 2) the nuclear magnetic resonance spectrum of the compound 1 H / 13 C NMR, 400 MHz) characteristics:

[0162] In DMSO as solvent, each peak is assigned as follows: 1 HNMR (400 MHz DMSO) δ: 8.57 (d, J = 6.72 Hz, 2H, -Ph), 8.17 (d, J = 7.00 Hz, 1H, -Ph), 7.92 (d, J = 6.88 Hz, 2H, -Ph), 7.80 (t, J = 8.24 Hz, 1H, -Ph), 7.62-7.71 (m, 3H, -Ph), 7.48 (t, J = 7.60 Hz, 1H, -Ph), 7.36 (t, J = 7.72 Hz, 1H, -Ph), 7.15 (t, J = 7.52 Hz, 1H, -Ph), 5.57 (s, 2H, -CH2-), 4.88 (t, J = 7.16 Hz, 2H, -CH2-), 4.43 (t, J = 6.60 Hz, 2H, -CH2-), 3.24-3.26 (m, 6H, -CH2-), 3.17 (t, J = 6.68 Hz, 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 = 7.28 Hz, 3H, -CH3), 0.82 (t, J = 7.28 Hz, 3H, -CH3); 13 C NMR (100 MHz 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, 120.37, 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.

[0163] Example 27 Compound 23

[0164] Compound 23 was synthesized by the method described in Example 5, and the physical and chemical properties of compound 23 are as follows:

[0165] 1) yellow solid;

[0166] 2) the nuclear magnetic resonance spectrum of the compound 1 H / 13 C NMR, 400 MHz) characteristics:

[0167] in DMSO as solvent, in which each peak is attributed to: 1 H NMR (400 MHz DMSO) δ: 8.62 (d, J = 5.56 Hz, 1H, -Ph), 8.58 (d, J = 6.92 Hz, 2H, -Ph), 8.17 (dd, J = 7.92, 1.12 Hz, 1H, -NH-), 7.90 (d, J = 7.00 Hz, 2H, -Ph), 7.79 (td, J = 8.32, 1.36 Hz, 1H, -Ph) 7.62-7.71 (m, 3H, -Ph), 7.48 (t, J = 7.08 Hz, 1H, -Ph), 7.36 (t, J = 7.32 Hz, 1H, -Ph), 7.15 (t, J = 7.44 Hz, 1H, -Ph), 5.23 (s, 2H, -CH2-), 4.87 (t, J = 7.24 Hz, 2H, -CH2-), 4.42 (t, J = 6.72 Hz, 2H, -CH2-), 3.25 (t, J = 7.20 Hz, 2H, -CH2-), 3.16 (t, J = 6.72 Hz, 2H, -CH2-), 3.12 (q, J = 6.76 Hz, 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 = 7.28 Hz, 3H, -CH3); 13 C NMR (100 MHz 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 Cl N5O2S [M-Cl] + calcd = 580.2741; found = 580.2744.

[0168] Compound 24 of Example 28

[0169] Compound 24 was synthesized by the method described in Example 5, and the physicochemical properties of compound 24 were as follows:

[0170] 1) yellow solid;

[0171] 2) the nuclear magnetic resonance spectrum of the compound (H NMR, 400MHz) was characterized as follows: 1 H / 13 C NMR, 400MHz) was characterized as follows:

[0172] In DMSO, each peak was assigned as follows: 1 H NMR (400MHz DMSO) δ: 8.70 (t, J = 5.36 Hz, 1H, -Ph), 8.59 (d, J = 6.96 Hz, 2H, -Ph), 8.16 (dd, J = 7.84, 1.04 Hz, 1H, -NH-), 7.91 (d, J = 7.00 Hz, 2H, -Ph), 7.79 (t, J = 6.96 Hz, 1H, -Ph) 7.61-7.70 (m, 3H, -Ph), 7.48 (t, J = 7.28 Hz, 1H, -Ph), 7.35 (t, J = 7.36 Hz, 1H, -Ph), 7.15 (t, J = 7.44 Hz, 1H, -Ph), 5.25 (s, 2H, -CH2-), 4.87 (t, J = 7.20 Hz, 2H, -CH2-), 4.42 (t, J = 6.72 Hz, 2H, -CH2-), 3.25 (t, J = 7.24 Hz, 2H, -CH2-), 3.16 (t, J = 6.72 Hz, 2H, -CH2-), 3.11 (q, J = 6.76 Hz, 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 = 6.72 Hz, 3H, -CH3); HRMS (ESI) C 35 H 40 ClN5O2S[M-Cl] + calcd = 594.2897; found = 594.2906.

[0173] Compound 25 of Example 29

[0174] Compound 25 was synthesized by the method described in Example 5, and the physicochemical properties of compound 25 were as follows:

[0175] 1) yellow solid;

[0176] 2) The NMR spectrum of the compound ( 1 H / 13 C NMR, 400 MHz) characteristics:

[0177] With DMSO as solvent, the peaks are attributed to: 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.

[0178] Example 30 Compound 26

[0179] Compound 26 was synthesized using the method described in Example 5. The physicochemical properties of compound 26 are as follows:

[0180] 1) yellow solid;

[0181] 2) the nuclear magnetic resonance spectrum of the compound 1 H / 13 C NMR, 400 MHz) characteristics:

[0182] in DMSO as solvent, in which each peak is attributed to: 1 HNMR (400 MHz DMSO) δ: 8.56 (d, J = 6.68 Hz, 2H, -Ph), 8.16 (d, J = 6.84 Hz, 1H, -Ph), 7.93 (d, J = 7.00 Hz, 2H, -Ph), 7.79 (t, J = 6.92 Hz, 1H, -Ph), 7.62-7.70 (m, 3H, -Ph), 7.48 (t, J = 7.20 Hz, 1H, -Ph), 7.36 (t, J = 7.28 Hz, 1H, -Ph), 7.15 (t, J = 7.40 Hz, 1H, -Ph), 5.62 (s, 2H, -CH2-), 4.87 (t, J = 7.24 Hz, 2H, -CH2-), 4.42 (t, J = 6.72 Hz, 2H, -CH2-), 3.70 (t, J = 4.16 Hz, 2H, -CH2-), 3.61 (t, J = 4.36 Hz, 2H, -CH2-), 3.45-3.48 (m, 4H, -CH2-), 3.26 (t, J = 7.24 Hz, 2H, -CH2-), 3.16 (t, J = 6.68 Hz, 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 (100 MHz DMSO) 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 Cl N5O3S [M-Cl] + calcd = 594.2533; found = 594.2542.

[0183] Example 31 Compound 27

[0184] Compound 27 was synthesized by the method described in Example 5. The physico-chemical properties of compound 27 are as follows:

[0185] 1) yellow solid;

[0186] 2) the nuclear magnetic resonance spectrum of the compound 1 H / 13 C NMR, 400MHz) characteristics:

[0187] In DMSO as solvent, each peak is assigned as follows: 1 HNMR (400MHz DMSO) δ: 8.57 (d, J = 6.92 Hz, 2H, -Ph), 8.16 (dd, J = 7.92, 1.12 Hz, 1H, -Ph), 7.94 (d, J = 7.08 Hz, 2H, -Ph), 7.79 (td, J = 8.32, 1.36 Hz, 1H, -Ph), 7.62-7.71 (m, 3H, -Ph), 7.48 (t, J = 7.16 Hz, 1H, -Ph), 7.36 (t, J = 7.32 Hz, 1H, -Ph), 7.15 (t, J = 7.36 Hz, 1H, -Ph), 5.61 (s, 2H, -CH2-), 4.88 (t, J = 7.08 Hz, 2H, -CH2-), 4.42 (t, J = 6.72 Hz, 2H, -CH2-), 3.71-3.74 (m, 4H, -CH2-), 3.26 (t, J = 7.28 Hz, 2H, -CH2-), 3.16 (t, J = 6.72 Hz, 2H, -CH2-), 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 (100MHz DMSO) 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.

[0188] Compound 28 of Example 32

[0189] Compound 28 was synthesized by the method described in Example 5. The physicochemical properties of compound 28 are as follows:

[0190] 1) yellow solid;

[0191] 2) the nuclear magnetic resonance spectrum of the compound 1 H / 13 C NMR, 400MHz) characteristics:

[0192] In DMSO as solvent, each peak is assigned as follows: 1 HNMR (400MHz DMSO) δ: 8.56 (d, J = 6.32 Hz, 2H, -Ph), 8.17 (dd, J = 7.88, 1.08 Hz, 1H, -Ph), 7.93 (d, J = 7.08 Hz, 2H, -Ph), 7.79 (t, J = 6.96 Hz, 1H, -Ph), 7.62-7.70 (m, 3H, -Ph), 7.48 (t, J = 7.20 Hz, 1H, -Ph), 7.36 (t, J = 7.32 Hz, 1H, -Ph), 7.15 (t, J = 7.40 Hz, 1H, -Ph), 5.50 (s, 2H, -CH2-), 4.87 (t, J = 7.16 Hz, 2H, -CH2-), 4.42 (t, J = 6.68 Hz, 2H, -CH2-), 3.50 (t, J = 6.76 Hz, 2H, -CH2-), 3.26 (t, J = 7.24 Hz, 4H, -CH2-), 3.16 (t, J = 6.68 Hz, 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.

[0193] Compound 29 of Example 33

[0194] Compound 29 was synthesized by the method described in Example 5. The physico-chemical properties of compound 29 are as follows:

[0195] 1) yellow solid;

[0196] 2) the nuclear magnetic resonance spectrum of the compound 1 H / 13 C NMR, 400MHz) characteristics:

[0197] In DMSO as solvent, each peak is assigned as follows: 1 HNMR (400MHz DMSO) δ: 8.56 (d, J = 6.72 Hz, 2H, -Ph), 8.16 (dd, J = 7.84, 0.96 Hz, 1H, -Ph), 7.92 (d, J = 6.80 Hz, 2H, -Ph), 7.79 (td, J = 8.32, 1.40 Hz, 1H, -Ph), 7.62-7.70 (m, 3H, -Ph), 7.53-7.57 (m, 4H, -Ph) 7.48 (t, J = 7.00 Hz, 2H, -Ph), 7.35 (t, J = 7.32 Hz, 1H, -Ph), 7.15 (t, J = 7.48 Hz, 1H, -Ph), 5.16 (s, 2H, -CH2-), 4.87 (t, J = 7.12 Hz, 2H, -CH2-), 4.42 (t, J = 6.68 Hz, 2H, -CH2-), 3.25 (t, J = 7.24 Hz, 2H, -CH2-), 3.23 (s, 3H, -CH3), 3.16 (t, J = 6.72 Hz, 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, 125.97, 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.

[0198] Compound 30 of Example 34

[0199] Compound 30 was synthesized by the method described in Example 5. The physicochemical properties of compound 30 are as follows:

[0200] 1) yellow solid;

[0201] 2) the nuclear magnetic resonance spectrum of the compound 1 H / 13 C NMR, 400MHz) characteristics:

[0202] In DMSO as solvent, each peak is assigned as follows: 1 HNMR (400MHz DMSO) δ: 8.73 (d, J = 6.60 Hz, 1H, -Ph), 8.59 (d, J = 6.92 Hz, 2H, -Ph), 8.16 (d, J = 6.88 Hz, 1H, -NH-), 7.90 (d, J = 7.04 Hz, 2H, -Ph), 7.79 (t, J = 7.00 Hz, 1H, -Ph), 7.62-7.70 (m, 3H, -Ph), 7.48 (t, J = 7.16 Hz, 1H, -Ph), 7.36 (t, J = 7.32 Hz, 1H, -Ph), 7.15 (t, J = 7.36 Hz, 1H, -Ph), 5.22 (s, 2H, -CH2-), 4.87 (t, J = 7.28 Hz, 2H, -CH2-), 4.42 (t, J = 6.68 Hz, 2H, -CH2-), 3.97-4.05 (m, 1H, -CH-), 3.25 (t, J = 7.24 Hz, 2H, -CH2-), 3.16 (t, J = 6.68 Hz, 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, 121.99, 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.

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

[0204] 1. Test bacteria:

[0205] Staphylococcus aureus ATCC 29213; Escherichia coli ATCC 25922; Methicillin-resistant Staphylococcus aureus (MRSA).

[0206] 2. Samples and reagents:

[0207] The samples were evodiamine, vancomycin, meropenem and compounds 1-30 prepared in the examples.

[0208] 3. Test method:

[0209] According to the standard of the Clinical and Laboratory Standards Institute (CLSI), the in vitro antibacterial activity of evodiamine and compounds 1-30 of the present application and the clinical antibacterial drug vancomycin were tested by using the micro-broth dilution method, and the minimum concentration of the drug for completely clear wells was observed by naked eye as the MIC value.

[0210] Table 1. MIC values of pyridinium salt-containing evodiamine derivatives 1-30 of the present application against 10 clinical isolates of MRSA and MRSA standard strain N315

[0211]

[0212]

[0213] S.a a : S. aureus ATCC 29213; M11-23 b : 10 clinical MRSA strains; E.c c : E. coli ATCC 25922; SI d : Selectivity index (HC 50 / MICs of S. aureus); ND e : Not detected; Van f : Vancomycin; MEM g : Meropenem. The experiment was repeated at least 3 times.

[0214] As can be seen from Table 1, the pyridinium salt-containing evodiamine derivatives prepared in the present application have significantly enhanced activity against gram-positive bacteria compared to the substrate evodiamine. The synthesized compounds have no significant antibacterial effect on E. coli. Compound 14 is the most active compound against gram-positive bacteria among the 30 pyridinium salt-containing evodiamine derivatives, with a MIC value of 0.5-2 μg / mL. Moreover, compound 14 has a relatively safe hemolytic activity (HC 50= 282.63 / 277.23) and good membrane selectivity and safety (SI = 565.26).

[0215] Example 2: Time-kill kinetics experiment of compound 14:

[0216] 1. Test bacteria:

[0217] S. aureus ATCC 29213; MRSA-11 (clinical isolate).

[0218] 2. Samples and reagents:

[0219] Samples: vancomycin and compound 14 prepared in the examples.

[0220] 3. Test method:

[0221] Single colonies of S. aureus ATCC 29213 and MRSA-11 were inoculated in 1 mL of MHB broth and incubated overnight. 2 μL of the bacterial solution was inoculated in 2 mL of MHB broth and incubated for 2.5 h (37 °C, 200 rpm). The concentration of the bacteria was determined to be 1 x 10 7 CFU / mL. Four groups of experiments were set up, i.e. 4 x MIC / 8 x MIC compound group, 8 x MIC vancomycin group, Control group. Compound 14 solution dissolved in DMSO and vancomycin solution were added to the cultured bacterial solution, and the Control group was added with the same amount of DMSO. Then the incubation was continued in a shaking incubator. 100 μL of the bacterial solution was taken at 0, 0.5, 1, 2, 4, 6, 8 h, respectively, washed and resuspended, and then gradient diluted in a 96-well plate. 10 μL of the sample dilution at different concentrations was taken and dropped onto MHA solid culture dishes, dried, and inverted in a 37 °C incubator for 16-24 h. The number of colonies at different time points was read and the killing curve was drawn. Three parallel controls were set up for each concentration. The experiment was repeated three times. The results are shown in Figures A and B. Figure 1 A, B.

[0222] Figure 1A, B show that the bactericidal rate of compound 14 on MRSA-11 and S. aureus ATCC29213 is higher than that of vancomycin at the same 8xMIC concentration. For compound 14, the CFU / mL of MRSA-11 is reduced by more than 3 logs (killing 99.9% of bacteria) within 4h at 8xMIC and within 4h at 4xMIC. In addition, the bacterial growth is also significantly inhibited at the concentration of 4xMIC of compound 14. Compound 14 shows rapid bactericidal activity on S. aureus ATCC29213, which can completely kill the bacteria within 4h at 8xMIC. These results show that compound 14 has rapid bactericidal activity on gram-positive bacteria MRSA-11 and S. aureus ATCC29213.

[0223] Example 3: Drug resistance induction experiment of compound 14:

[0224] After determining the initial MIC value of target compound 14 and positive control drug norfloxacin by drug sensitivity test, single colony of S. aureus ATCC 29213 was picked and placed in two 1mL MHB liquid culture medium, and the test compound and norfloxacin were added respectively to make the final concentration be sub-inhibitory concentration, and cultured for 12h. The bacterial liquid was taken with a bacterial loop, and the bacterial liquid was inoculated into 1 / 2MIC target compound and positive control drug-containing MHA solid culture medium and drug-free solid culture medium by three-zone streaking method. After the single colony appeared, the MIC value was re-determined and recorded. The drug-free plate was the first generation MIC value, and the drug-containing plate was the second generation MIC value. The above operation was repeated, and the sub-inhibitory concentration was adjusted according to the MIC value of each generation. A total of 20 generations were cultured. The results are shown in Table 2. Figure 1 C.

[0225] From Figure 1 C results show that S. aureus ATCC29213 fails to produce drug-resistant mutants within 20 days in the presence of sub-inhibitory concentration (1 / 2MIC) of compound 14, indicating that the target is non-specific. After 4 generations, the MIC of norfloxacin begins to rise, and drug resistance appears. After 18 generations, the MIC of norfloxacin increases by 128 times. It shows that the low drug resistance of compound 14 is beneficially affected by its rapid bactericidal activity.

[0226] Example 4: In vivo safety evaluation experiment - subcutaneous injection

[0227] 1. Reagents:

[0228] Compound 14 prepared in the examples, 0.9% NaCl.

[0229] 2. Test animals

[0230] SPF grade KM female mice (purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., body weight 20-22 g, 7-8 weeks old).

[0231] 3. Test method

[0232] After compound 14 was dissolved with DMSO, physiological saline was used to prepare test compound solutions with concentrations of 80, 40, 20, 10, and 5 mg / kg. Twenty-four healthy female KM mice with consistent body weight were selected and randomly divided into six groups, including five experimental groups and one blank control group, with four mice in each group. The mice in the experimental groups were subcutaneously injected with 60 μL of compound solutions with different concentrations on the back, and the mice in the blank control group were subcutaneously injected with 60 μL of physiological saline on the back. The survival state of the mice and whether the skin showed irritant reactions were observed. After 24 h, the mice with the maximum injection concentration that did not show death, hair loss, and other irritant reactions were subjected to eyeball blood sampling for blood routine test and liver and kidney function test to observe whether there were significant changes compared with the blank control group.

[0233] The results showed that when the concentration of compound 14 was ≤20 mg / kg, no adverse reactions (such as redness, hardness, and ulceration) occurred in the skin of the mice after subcutaneous injection of the compound with different concentrations. Therefore, blood routine test and blood biochemical index test were performed on the mice with this dose. Figure 2 The blood samples were subjected to blood routine test and blood biochemical 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 there was no significant difference in each result after subcutaneous injection of compound 14 (20 mg / kg) into KM mice compared with the blank control group (0.9% NaCl), indicating that compound 14 had certain in vivo safety.

[0234] Application Example 5: In vivo anti-MRSA infection activity experiment of compound 14—subcutaneous injection

[0235] 1. Test bacteria:

[0236] MRSA-11 (clinical isolate)

[0237] 2. Samples and reagents:

[0238] The samples were vancomycin and compound 14 prepared in the examples, and 0.9% NaCl.

[0239] 3. Test animals:

[0240] SPF grade KM mice (purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., body weight 20-22 g, 7-8 weeks old).

[0241] 4. Test method:

[0242] Thirty healthy female KM mice with uniform body weight were depilated on the back, 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 back of the mice was disinfected, 60 μL of MRSA-11 bacterial suspension with a concentration of 6×10 8 CFU / mL was subcutaneously injected to the mice in each group except the blank group. After 2 h, 60 μL of the compound with different contents, vancomycin, or normal saline was injected in situ. The survival and skin infection of the mice in each group were recorded. After 24 h of administration, the mice in each group were sacrificed by dislocation, and the skin in the infected area was taken. The skin tissue in the infected area was ground to a broken state using a tissue grinder, and the skin tissue grinding liquid was sucked. The bacterial colony count was read using a dipstick counting method. The results are shown in Table 1. Figure 3

[0243] The results show that the bacterial load in the skin tissue of the mice in the Control group is about 6.9 log 10 CFU / g, and the number of MRSA cells in the skin of the mice is significantly reduced after treatment with low-dose 14 (5 mg / kg) and vancomycin (5 mg / kg) (P<0.0001), and the bacterial load is reduced by 1.8 log 10 CFU / g, 1.4 log 10 CFU / g, respectively, showing a moderate treatment effect. After treatment with 14 (10 mg / kg), the bacterial load in the skin is reduced by about 5.4 log 10 CFU / g, and the treatment effect is the most significant. Therefore, it is confirmed that the evodiamine derivative 14 containing a pyridine quaternary ammonium salt has a good treatment effect on MRSA-infected mouse skin abscesses, and the treatment effect is the same as that of the control drug vancomycin at the same dose. The compound 14 is expected to be developed into a new drug for resisting MRSA.

[0244] Application Example 6: In vivo safety evaluation experiment - intraperitoneal injection

[0245] 1. Reagents:

[0246] The compound 14 prepared in the example, corn oil.

[0247] 2. Test animals

[0248] SPF grade BALB / c female mice (purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., body weight 16-18 g, 7-8 weeks old). ​

[0249] 3. Test method

[0250] Compound 14 was dissolved in DMSO and then prepared into test compound solutions with concentrations of 40, 20, 15, 10, and 5 mg / kg using corn oil. Thirty-six healthy female BALB / c mice with uniform body 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 groups were injected intraperitoneally with 100 μL of compound solutions with different concentrations, and the mice in the blank control group were injected subcutaneously with 100 μL of corn oil on the back, and then injected intraperitoneally with drugs every 12 h, for two consecutive times. The survival state and survival rate of the mice were recorded for seven consecutive days from the first day of administration, and food and water were normally provided during the period. After the maximum tolerated dose was determined, the mice were administered again, and blood was taken from the eyeballs 24 h later for blood routine test and liver and kidney function test to observe whether there were significant changes compared with the blank control group.

[0251] The results showed that, by injecting different concentrations of compound into the abdominal cavity of the mice, when the concentration of compound 14 was ≤15 mg / kg, the skin of the mice did not show any adverse reactions (such as death, listlessness, and tangled hair, etc.), and therefore the blood routine test and blood biochemical index test were performed on the mice at this dose. Figure 4 The blood samples were subjected to blood routine test and blood biochemical 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 the BALB / c mice were injected intraperitoneally with compound 14 (15 mg / kg), there was no significant difference in each result, indicating that compound 14 had certain in vivo safety.

[0252] Application Example 7: In vivo anti-MRSA infection activity experiment of compound 14—intraperitoneal injection

[0253] 1. Test bacteria:

[0254] MRSA-11 (clinical isolate)

[0255] 2. Samples and reagents:

[0256] The samples were: vancomycin, compound 14 prepared in the examples, corn oil, and 0.9% NaCl.

[0257] 3. Test animals:

[0258] SPF-level BALB / c female mice (purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., body weight 16-18 g, 7-8 weeks old).

[0259] 4. Test method:

[0260] 4.1 Determination of sublethal and lethal bacterial counts

[0261] 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. 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 counted on a plate to calculate the bacterial load. Figure 5 shown.

[0262] 4.2 Organ bacterial burden and survival rate experiments

[0263] 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 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 compound (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 skin tissue grinding fluid from 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.

[0264] 30 SPF level BALB / c female mice with consistent body size and weight were randomly divided into blank group, control group, vancomycin group (12 mg / kg), low-dose compound group (6 mg / kg), and high-dose compound group (12 mg / kg), with 6 mice in each group. The blank group was not given any drug, and the other groups were injected with 100 μL of normal saline containing a lethal amount of bacteria. After 0.5 h, the control group was injected intraperitoneally with 100 μL of corn oil, and the two groups were injected with 100 μL of compound (6 mg / kg, 12 mg / kg), and one group was injected with positive control drug vancomycin (12 mg / kg), with administration every 12 h, and continuous administration for two times. Continuous observation was carried out for seven days, the survival state of the mice was recorded, the mortality rate was calculated, and the results are shown in Table 1. Figure 7 Table 1: Survival rate of mice in different groups

Claims

1. A quaternary ammonium pyridinium salt-containing 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; the heteroatoms in ring A, except the N atom connected to the parent nucleus, are absent or selected from one of N, O and S.

2. 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, 3. The method for preparing the quaternary pyridinium ammonium salt-containing evodia rutaecarpine derivative according to claim 1 or 2, characterized in that: The following steps are involved: (1) Using rutaecarpine 1 as a substrate, it reacts with dibromoalkane 5 to obtain monobromoalkane-substituted intermediate 2; (2) Intermediate 2 reacts with 4-mercaptopyridine to produce 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 claim 1 or 2.

4. The method for preparing the quaternary pyridinium ammonium salt-containing evodia rutaecarpine derivative according to claim 3, characterized in that: In step (1), the reaction molar ratio of the rutaecarpine 1 to the dibromoalkane 5 is 1:(7.0-12.0), the catalyst is potassium carbonate, the reaction molar ratio of the rutaecarpine 1 to potassium carbonate is 1:(6.5-7.0), the reaction temperature is 45-50°C, and the reaction solvent is acetone.

5. The method for preparing the rutaecarpine derivative containing a quaternary pyridinium ammonium salt according to claim 3, characterized in that: In step (2), the monobromoalkane-substituted intermediate 2 reacts with 4-mercaptopyridine in a molar ratio of 1:(1.0-1.5), the catalysts are potassium carbonate and potassium iodide, the molar ratio of the intermediate 2 to potassium carbonate is 1:(1.0-1.7), and the molar ratio of the intermediate 2 to potassium iodide is 1:(0.01-0.03), the reaction temperature is 45-50°C, and the reaction solvent is acetone.

6. The method for preparing the quaternary pyridinium ammonium salt-containing evodia rutaecarpine derivative according to claim 3, characterized in that: In step (3), the preparation method of chloroacetamide 6 comprises the following steps: chloroacetyl chloride and aliphatic amine / aromatic amine containing different substituents undergo substitution reaction under alkaline catalyst conditions to generate chloroacetamide 6:

7. The method for preparing the quaternary pyridinium ammonium salt-containing evodia rutaecarpine derivative according to claim 6, characterized in that: 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 quaternary pyridinium ammonium salt-containing evodiamine derivative 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 quaternary pyridinium ammonium salt-containing evodiamine derivative according to claim 1 or 2 in the preparation of antibacterial drugs, characterized in that: The antibacterial drug can inhibit Staphylococcus aureus or methicillin-resistant Staphylococcus aureus.

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