Heterocyclic piperazine trichothecene derivatives and uses thereof

Abstract

The application discloses a heterocyclic piperazine truncated pleuromutilin derivative and application thereof, relates to the field of piperazine truncated pleuromutilin derivatives and application thereof, and the heterocyclic piperazine truncated pleuromutilin derivative comprises a compound or a stereoisomer or a pharmaceutically acceptable salt thereof as shown in a structural formula of formula I.The heterocyclic piperazine truncated pleuromutilin derivative of the application shows excellent antibacterial effects on gram-positive bacteria, such as Staphylococcus aureus drug-resistant strains ATCC 33591 and ATCC 43300, Staphylococcus aureus sensitive strains ATCC 29213, Staphylococcus epidermidis drug-resistant strains ATCC 51625 and Staphylococcus epidermidis sensitive strains ATCC 12228.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of antibacterial drugs, in particular to a heterocyclic piperazine pleuromutilin derivative and its use. BACKGROUND

[0002] The widespread use and even abuse of antibiotics has led to the emergence of increasingly serious drug resistance in many bacteria. It is estimated that the number of deaths caused by antibiotic resistance worldwide has reached 100 million per year, causing economic losses of up to 100 trillion US dollars. In particular, methicillin-resistant Staphylococcus aureus (MRSA) infection has led to higher morbidity and mortality, resulting in longer treatment times and higher treatment costs, which has attracted widespread attention from researchers. Therefore, it is increasingly important to explore and discover new drugs against drug-resistant bacteria.

[0003] Pleuromutilin is a natural product with antibacterial activity discovered in the 1950s, which is a class of diterpenoids with a tricyclic skeleton produced by deep culture of higher fungi Pleurotsmutilus and Pleurots passeckerianus species. Studies have shown that modification of C14 position of pleuromutilin can produce compounds with stronger antibacterial activity. After modification at C14, Tiamulin and Valnemulin were approved for use in poultry and pigs as veterinary antibiotics in 1979 and 1999, respectively. In 2007, Retapamulin was approved as the first human drug for the treatment of skin impetigo, small area skin damage infection and secondary infection mainly caused by Staphylococcus aureus. On August 19, 2019, Lefamulin was approved by FDA for intravenous and oral treatment of community-acquired bacterial pneumonia (CABP).

[0004]

[0005] Gastrointestinal side effects, liver toxicity, and challenges in compound synthesis are all factors that hinder the use of pleuromutilin as a human oral or injection preparation. Based on the above factors, researchers have made a lot of derivatization and modification of pleuromutilin. In 1982, Azamulin entered clinical research due to its excellent in vitro antibacterial activity, but due to the strong toxicity of the compound, it can inhibit the normal metabolism of human liver microsomes (CYP3A4, IC 50 =0.03-0.24 μ M, Drug Metab. Dispos. , 2004, 105). In addition, Hugues Chanteux et al. Drug Metab. Dispos., 2020, 778) found that Azamulin (3 μ M) can significantly inhibit CYP3A4 (>90%), which limits the subsequent clinical application of the compound. SUMMARY

[0006] The purpose of the present application is to provide a heterocyclic piperazine truncated side ear derivative and its use, to solve the problem that the truncated side ear and its derivatives in the prior art will inhibit the normal metabolism of human liver microsomes in the process of clinical application.

[0007] First, the embodiment of the present application provides a heterocyclic piperazine truncated side ear derivative, including a compound or its stereoisomer or a pharmaceutically acceptable salt of the structural formula as shown in formula I,

[0008]

[0009] Formula I

[0010] Among them, R is selected from 、 、 、 and .

[0011] As an optional implementation, when R is , R1 is selected from one of H, F, Cl, Br, CH3, OCH3, NO2 and NHBoc, and R2 is selected from one of H, F, Cl, Br, CH3 or OCH3.

[0012] As an optional implementation, when R is , R3 is H.

[0013] As an optional implementation, when R is , R4 is H.

[0014] As an optional implementation, when R is ,

[0015] R5 is selected from H or CH3, R6 is H, R7 is H, and R8 is H;

[0016] or R6 is selected from H or CH3, R5 is H, R7 is CH3 or H, and R8 is H;

[0017] or R7 is selected from one of NO2, Cl and Br, and R5, R6 and R8 are each selected from one of H, Cl and Br.

[0018] As an optional implementation, the compound includes one of the following structures:

[0019] 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 .

[0020] As an optional implementation, the pharmaceutically acceptable salt comprises one of hydrochloride, fumarate, malate, hydrobromide, succinate, phosphate, methanesulfonate and benzoate.

[0021] Secondly, the embodiment of the present application further provides a heterocyclic piperazine truncated side ear derivative, comprising the heterocyclic piperazine truncated side ear derivative for treating infectious diseases.

[0022] As an optional implementation, the infectious disease comprises mycoplasma or drug-resistant bacteria.

[0023] As an optional implementation, the heterocyclic piperazine truncated side ear derivative is combined with a pharmaceutically acceptable adjuvant before administration.

[0024] It should be noted that the "pharmaceutically acceptable salt" refers to a salt of the present application compound which maintains the bioavailability and characteristics of the free acid or free base, and the free acid is obtained by reacting with a non-toxic organic base or inorganic base, and the free base is obtained by reacting with a non-toxic inorganic acid or organic acid. "Pharmaceutically acceptable adjuvant" refers to an inert substance added to a pharmaceutical composition to facilitate the administration of the compound.

[0025] Compared with the prior art, the embodiment of the present application has the following advantages and beneficial effects:

[0026] The piperazine truncated pleuromutilin derivative provided by the embodiment of the present application has novel structure, good antibacterial activity and low inhibition to CYP3A4. The derivative shows excellent antibacterial effect on Gram-positive bacteria, such as Staphylococcus aureus resistant strain ATCC 33591 and ATCC 43300, Staphylococcus aureus sensitive strain ATCC 29213, Staphylococcus epidermidis resistant strain ATCC 51625 and Staphylococcus epidermidis sensitive strain ATCC 12228. BRIEF DESCRIPTION OF DRAWINGS

[0027] In order to more clearly illustrate the technical solutions of the exemplary embodiments of the present application, the drawings needed in the embodiments will be briefly introduced as follows. It should be understood that the following drawings only show some embodiments of the present application, and therefore should not be considered as a limitation to the scope. Other related drawings can also be obtained by those skilled in the art without creative effort. In the drawings:

[0028] Figure 1 is a bactericidal curve diagram, wherein Figure 1 A is a time-killing curve diagram of tiamulin on MRSA ATCC 33591, Figure 1 B is a time-killing curve diagram of compound 6a on MRSA ATCC 33591;

[0029] Figure 2 is a diagram of the inhibitory effect of compound 6a at different concentrations on CYP3A4 enzyme;

[0030] Figure 3 is a diagram of the bacterial load in the thigh of mice after treatment with 40 mg / kg of compound 6a and retapamulin after MRSA ATCC 33591 infection, wherein "*" represents a significant difference, and "**" represents a very significant difference;

[0031] Figure 4 is the therapeutic effect of compound 6a in a mouse systemic infection model;

[0032] Figure 5 is the therapeutic effect of tiamulin in a mouse systemic infection model;

[0033] Figure 6 is an analysis of the bacterial load in the main organs of systemic MRSA ATCC 33591 infected mice after treatment with compound 6a and tiamulin (40 mg / kg), wherein Figure 6 A corresponds to the organ "heart", Figure 6 B corresponds to the organ "liver", Figure 6 C corresponds to the organ "spleen", Figure 6 D corresponds to the organ "lung", Figure 6 E corresponds to the organ "kidney".

[0034] Figure 7 H&E stained organ tissue section results for MRSA ATCC 33591 infected mice. DETAILED DESCRIPTION

[0035] In order to make the objects, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be described clearly and completely below. Obviously, the described embodiments are some but not all of the embodiments of the present application.

[0036] Therefore, the detailed description of the embodiments of the present application provided below is not intended to limit the scope of the claimed application, but merely represents the selected embodiments of the application. Based on the embodiments in the present application, all other embodiments obtained by a person of ordinary skill in the art without creative work fall within the scope of protection of the present application.

[0037] Example 1: The synthetic route of compound 4 is shown as follows:

[0038]

[0039] The steps for synthesizing compound 2 are shown as follows:

[0040] p-toluenesulfonyl chloride (4.3 g, 22.7 mmol) and pleuromutilin (7.8 g, 20.6 mmol) were dissolved in a mixed solution of 25 mL methyl tert-butyl ether and water (v / v=4:1), and 5 mL sodium hydroxide solution (10 M) was slowly added dropwise to the above mixed solution under ice bath conditions. Then, the device was placed in heating reaction at 60 ℃ for about 1 h. After the reaction was completed, it was poured into a beaker containing an appropriate amount of water, and the filter cake was washed with water to obtain a white solid, which was dried to obtain the target product 2 (yield 92.3%).

[0041] 1 H NMR (400 MHz, CDCl3): δ (ppm) 7.81 (d, J = 8.4 Hz, 2H), 7.35 (d, J =8.4 Hz, 2H), 6.41 (dd, J = 17.2, 11.2 Hz, 1H), 5.76 (d, J = 8.4 Hz, 1H), 5.33(dd, J = 11.2, 1.2 Hz, 1H), 5.19 (dd, J= 17.2, 1.2 Hz, 1H), 4.48 (s, 2H), 3.34(d, J = 6.4 Hz, 1H), 2.45 (s, 3H), 2.33 – 1.99 (m, 5H), 1.81 – 1.41 (m, 6H), 1.40 (s, 3H), 1.38 – 1.30 (m, 1H), 1.29 – 1.20 (m, 1H), 1.15 (s, 3H), 1.13 –1.05 (m, 1H), 0.87 (d, J = 6.8 Hz, 3H), 0.62 (d, J = 6.8 Hz, 3H).

[0042] The steps for synthesizing compound 3 are as follows:

[0043] Pleurotus sulfonate (532.70 mg, 1.00 mmol) and NaI (15.00 mg, 0.10 mmol) were dissolved in dry acetonitrile solution and refluxed for 0.5 h. Then, anhydrous piperazine (172.30 mg, 2.00 mmol) and K₂CO₃ (276.40 mg, 2.00 mmol) were added to the solution, and the mixture was stirred and refluxed. The reaction progress was monitored by TLC. After the reaction was complete, the mixture was cooled to room temperature, concentrated under reduced pressure, and purified by column chromatography to obtain a white powdery intermediate 3 (yield 75.2%).

[0044] 1 H NMR (400 MHz, CDCl3): δ (ppm) 7.74 (d, J = 8.0 Hz, 1H), 7.22 (d, J =8.0 Hz, 1H), 6.48 (dd, J = 17.2, 11.2 Hz, 1H), 5.78 (d, J = 8.4 Hz, 1H), 5.33(dd, J = 11.2, 1.2 Hz, 1H), 5.19 (dd, J = 17.2, 1.6 Hz, 1H), 3.35 (d, J= 6.0 Hz,1H), 3.20 – 3.02 (m, 6H), 2.79 – 2.62 (m, 4H), 2.40 – 1.99 (m, 7H), 1.81 –1.31 (m, 10H), 1.16 (s, 3H), 1.14 – 1.07 m, 1H), 0.87 (d, J = 6.8 Hz, 3H), 0.69(d, J = 6.8 Hz, 3H).

[0045] The synthesis of compound 4 is shown in the following steps:

[0046] Intermediate 3 (1.00 g, 2.24 mmol) was dissolved in 15 mL of dichloromethane, K2CO3(0.62 g, 4.48 mmol) was added to the reaction system, then chloroacetyl chloride (0.28 g, 2.46 mmol) was slowly added, and the reaction was carried out in an ice bath for about 3 hours. TLC was used to detect the progress of the reaction, and after the reaction was completed, an appropriate amount of 15% sodium chloride aqueous solution was added to the reaction liquid, extracted with chloroform (3 x 20.00 mL), and the organic phase was collected, dried with anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the white solid target compound 4.

[0047] 1 H NMR (400 MHz, CDCl3): δ (ppm) 6.49 (dd, J = 17.2, 11.2 Hz, 1H, H19),5.79 (d, J = 8.4 Hz, 1H, H14), 5.34 (dd, J = 11.2, 1.6 Hz, 1H, H20), 5.20 (dd, J=17.2, 1.6 Hz, 1H, H20), 4.05 (s, 2H, H28), 3.75 – 3.63 (m, 2H, Piperazinyl-H), 3.61 – 3.54 (m, 2H, Piperazinyl-H), 3.40 – 3.32 (m, 1H, H11), 3.26 – 3.16 (m, 1H, H22), 3.15 – 3.03 (m, 1H, H22), 2.76 – 2.50 (m, 4H, Piperazinyl-H), 2.40 – 2.03 (m, 6H, H2, H4, H10, H13), 1.83 – 1.33 (m, 9H, H1, H6, H7, H8,H15), 1.16 (s, 3H, H18), 1.15 – 1.06 (m, 1H, H8), 0.88 (d, J = 6.8 Hz, 3H,H17), 0.71 (d, J = 6.8 Hz, 3H, H16).

[0048] Example 2: Compound 5a The synthetic route is shown below:

[0049]

[0050] The specific synthetic steps included dissolving truncated pleurotin intermediate 4 (1.00 mmol), 4-hydroxycoumarin (1.10 mmol), and K2CO3 (2.00 mmol) in N,N-dimethylformamide (6.00 mL), respectively. o The reaction was stirred at C. After the reaction was complete, 100 mL of water was added, the filter cake was collected by vacuum filtration, and purified by column chromatography to obtain the target product.

[0051] Compound 5a is a white powder; yield: 60.7%; melting point: 112.7-114.3℃;

[0052] 1 H NMR (400 MHz, CDCl3): δ (ppm) 7.87 (dd, J = 8.0, 1.6 Hz, 1H, H36), 7.59 – 7.51 (m, 1H, H33), 7.35 – 7.26 (m, 2H, H34, H35), 6.49 (dd, J = 17.2,11.2 Hz, 1H, H19), 5.78 (d,J = 8.4 Hz, 1H, H14), 5.64 (s, 1H, H30), 5.34 (dd, J = 11.2, 1.2 Hz, 1H, H20), 5.19 (dd, J = 17.2, 1.2 Hz, 1H, H20), 4.86 (s, 2H,H2, 3.76 – 3.66 (m, 2H, Piperazinyl-H), 3.60 – 3.47 (m, 2H, Piperazinyl-H),3.38 – 3.31 (m, 1H, H11), 3.26 – 3.17 (m, 1H, H22), 3.14 – 3.03 (m, 1H, H22),2.70 – 2.54 (m, 4H, Piperazinyl-H), 2.39 – 1.98 (m, 6H, H2, H4, H10, H13),1.83 – 1.32 (m, 9H, H1, H6, H7, H8, H15), 1.16 (s, 3H, H18), 1.14 – 1.06 (m,1H, H8)), 0.88 (d, J = 6.8 Hz, 3H, H17), 0.70 (d, J = 6.8 Hz, 3H, H16). 13C NMR (101 MHz, CDCl3): δ (ppm) 217.1 (C3), 168.8 (C21), 164.9 (C29), 162.5 (C31),153.4 (C32), 139.1 (C19), 132.7 (C34), 124.1 (C35), 123.1 (C36), 117.3 (C20),116.8 (C37), 115.3 (C33), 91.5 (C30), 74.6 (C11), 68.6 (C14), 59.5(Piperazinyl-C), 58.2 (C4), 52.7 (Piperazinyl-C), 52.3 (Piperazinyl-C),45.5(C22), 45.0 (C9), 44.0 (C12,C13), 41.8 (C5), 36.7 (C6), 36.1 (C10), 34.5(C2), 30.4 (C8), 26.8 (C7), 26.4 (C18), 24.9 (C1), 16.8 (C16), 14.9 (C15),11.5 (C17).HRMS: calculated for C 37 H 48 N2O8([M + H] + ): 649.3483; found 649.3500.

[0053] Example 3: Synthesis of compound 5b was synthesized from compound 4, the specific synthesis steps are referred to Example 2.

[0054] Compound 5b is a white powder; yield: 35.7 %; melting point: 113.2-115.7 °C;

[0055] 1 H NMR (400 MHz, CDCl3): δ (ppm) 7.75 (d, J = 8.8 Hz, 1H, H36), 6.84(dd, J = 8.8, 2.4 Hz, 1H, H33), 6.80 – 6.76 (m, 1H, H35), 6.49 (dd, J = 17.2,11.2 Hz, 1H, H19), 5.79 (d, J = 8.4 Hz, 1H, H14), 5.50 (s, 1H, H30), 5.34 (dd,J = 11.2, 1.2 Hz, 1H, H20), 5.20 (dd, J = 17.2, 1.2 Hz, 1H, H20), 4.83 (s, 2H,H28), 3.86 (s, 3H, OMe-H), 3.75 – 3.66 (m, 2H, Piperazinyl-H), 3.57 – 3.47(m, 2H, Piperazinyl-H), 3.39 – 3.30 (m, 1H, H11), 3.27 – 3.17 (m, 1H, H22),3.14 – 3.06 (m, 1H, H22), 2.71 – 2.52 (m, 4H, Piperazinyl-H), 2.38 – 2.01 (m,6H, H2, H4, H10, H13), 1.81 – 1.31 (m, 9H, H1, H6, H7, H8, H15), 1.16 (s, 3H,H18), 1.14 – 1.06 (m, 1H, H8), 0.88 (d, J = 6.8 Hz, 3H, H17), 0.70 (d, J = 6.8Hz, 3H, H16). 13 C NMR (101 MHz, CDCl3):δ (ppm) 217.1 (C3), 168.8 (C21), 165.3(C29), 163.6 (C31), 163.0 (C34), 155.2 (C32), 139.1 (C19), 124.2 (C36), 117.3(C20), 112.4 (C35), 108.5 (C37), 100.5 (C33), 88.9 (C30), 74.6 (C11), 68.6(C14), 59.5 (Piperazinyl-C), 58.2 (C14), 55.8 (C38), 52.5 (Piperazinyl-C),45.5 (C22), 45.0 (C9), 44.0 (C12,C13), 41.8 (C5), 36.7 (C6), 36.1 (C10), 34.5(C2), 30.4 (C8), 26.8 (C7), 26.4 (C18), 24.9 (C1), 16.7 (C16), 14.9 (C15),11.5 (C17). HRMS: calculated for C38 H 50 N2O9([M + H] + ): 679.3589; found679.3589.

[0056] Example 4: Synthesis of compound 5c The synthesis of compound 5c was still synthesized from compound 4, the specific synthesis steps refer to Example 2.

[0057] Compound 5c is a white powder; yield: 40.6 %; melting point 117.6-120.2 °C;

[0058] 1 H NMR (400 MHz, CDCl3): δ (ppm) 7.33 – 7.27 (m, 1H, H33), 7.24 (s,1H, H34), 7.17 – 7.10 (m, 1H, H36), 6.49 (dd, J = 17.2, 11.2 Hz, 1H, H19), 5.79(d, J = 8.4 Hz, 1H, H14), 5.65 (s, 1H, H30), 5.34 (d, J = 11.6 Hz, 1H, H20), 5.21(d, J = 17.2 Hz, 1H, H20), 4.88 (s, 2H, H28), 3.85 (s, 3H, OMe-H), 3.78 – 3.68(m, 2H, Piperazinyl-H), 3.57 – 3.50 (m, 2H, Piperazinyl-H), 3.41 – 3.34 (m,1H, H11), 3.27 – 3.18 (m, 1H, H22), 3.16 – 3.05 (m, 1H, H22), 2.76 – 2.52 (m,4H, Piperazinyl-H), 2.42 – 2.00 (m, 6H, H2, H4, H10, H13), 1.88 – 1.33 (m,9H, H1, H6, H7, H8, H15), 1.17 (s, 3H, H18), 1.16 – 1.08 (m, 1H, H8), 0.89(d, J = 6.8 Hz, 3H, H17), 0.72 (d, J = 6.8 Hz, 3H, H16). 13C NMR (101 MHz, CDCl3): δ (ppm) 216.9 (C3), 168.8 (C21), 164.6 (C29), 162.6 (C31), 156.0 (C35), 147.8(C32), 139.1 (C19), 120.8 (C33), 117.9 (C37), 117.2 (C20), 115.7 (C34), 105.0(C36), 91.7 (C30), 74.6 (C11), 68.6 (C14), 59.5 (Piperazinyl-C), 58.2 (C4),55.9 (C38), 52.4 (Piperazinyl-C), 45.5 (C22), 45.5 (C9), 44.9 (C13), 44.0(C12), 41.9 (C5), 36.7 (C6), 36.1 (C10), 34.4 (C2), 30.4 (C8), 26.8 (C7),26.5 (C18), 24.9 (C1), 16.7 (C16), 14.9 (C15), 11.5 (C17). HRMS: calculated for C 38 H 50 N2O9([M + H] + ): 679.3589; found 679.3585.

[0059] Example 5: Synthesis of compound 5d was synthesized from compound 4, the specific synthesis steps are referred to Example 2.

[0060] Compound 5d is a white powder; yield: 73.5 %; melting point: 113.4-115.7 °C;

[0061] 1 H NMR (400 MHz, CDCl3): δ (ppm) 7.65 (s, 1H, H36), 7.35 (d, J = 8.4 Hz,1H, H35), 7.19 (d, J = 8.4 Hz, 1H, H33), 6.47 (dd, J = 17.2, 11.2 Hz, 1H, H19),5.77 (d, J = 8.4 Hz, 1H, H14), 5.61 (s, 1H, H30), 5.32 (d, J= 11.2 Hz, 1H, H20),5.19 (d, J = 17.2 Hz, 1H, H20), 4.84 (s, 2H, H28), 3.83 – 3.66 (m, 2H,Piperazinyl-H), 3.56 – 3.48 (m, 2H, Piperazinyl-H), 3.40 – 3.30 (m, 1H, H11),3.27 – 3.16 (m, 1H, H22), 3.14 – 3.03 (m, 1H, H22), 2.76 – 2.49 (m, 4H,Piperazinyl-H), 2.39 (s, 3H, Me-H), 2.35 – 1.96 (m, 6H, H2, H4, H10, H13),1.83 – 1.31 (m, 9H, H1, H6, H7, H8, H15), 1.15 (s, 3H, H18), 1.13 – 1.07 (m,1H, H8), 0.87 (d, J = 6.8 Hz, 3H, H17), 0.70 (d, J = 6.8 Hz, 3H, H16). 13 C NMR (101MHz, CDCl3):δ (ppm) 216.9 (C3), 168.8 (C21), 164.9 (C29), 163.5 (C31), 162.7(C32), 151.5 (C34), 139.1 (C19), 133.7 (C36), 122.7 (C35), 117.2 (C20), 116.5(C33), 115.0 (C37), 91.4 (C30), 74.6 (C11), 68.6 (C14), 59.5 (Piperazinyl-C),58.2 (C4), 52.5 (Piperazinyl-C), 45.5 (C22), 45.1 (C9), 44.0 (C12,C13), 41.8(C5), 36.7 (C6), 36.1 (C10), 34.4 (C2), 30.4 (C8), 26.8 (C7), 26.5 (C18),24.9 (C1), 20.9 (C38), 16.7 (C16), 14.9 (C15), 11.5 (C17).HRMS: calculatedfor C 38 H 50N2O8([M + H] + ): 663.3640; found 663.3647.

[0062] Example 6: Synthesis of compound 5e The synthesis of compound 5e was still synthesized from compound 4, the specific synthesis steps are referred to Example 2.

[0063] Compound 5e is a white powder; yield: 75.8 %; melting point: 122.8-124.2 °C;

[0064] 1 H NMR (400 MHz, CDCl3): δ (ppm) 7.73 (d, J = 8.0 Hz, 1H, H36), 7.15 –7.05 (m, 2H, H33, H34), 6.49 (dd, J = 17.2, 11.2 Hz, 1H, H19), 5.79 (d, J = 8.4Hz, 1H, H14), 5.58 (s, 1H, H30), 5.34 (dd, J = 11.2, 1.2 Hz, 1H, H20), 5.20(dd, J = 17.2, 1.2 Hz, 1H, H20), 4.83 (s, 2H, H28), 3.77 – 3.62 (m, 2H,Piperazinyl-H), 3.60 – 3.45 (m, 2H, Piperazinyl-H), 3.41 – 3.30 (m, 1H, H11),3.28 – 3.16 (m, 1H, H22), 3.15 – 3.05 (m, 1H, H22), 2.72 – 2.51 (m, 4H,Piperazinyl-H), 2.44 (s, 3H, Me-H), 2.38 – 2.00 (m, 6H, H2, H4, H10, H13),1.86 – 1.32 (m, 9H, H1, H6, H7, H8, H15), 1.16 (s, 3H, H18), 1.15 – 1.08 (m,1H, H8), 0.88 (d, J = 6.8, Hz, 3H, H17), 0.71 (d, J = 6.8 Hz, 3H, H16). 13C NMR (101 MHz, CDCl3): δ (ppm) 216.9 (C3), 168.8 (C21), 164.9 (C29), 163.5 (C31),162.6 (C32), 151.6 (C35), 139.1 (C19), 133.8 (C34), 122.7 (C36), 117.3 (C20),116.6 (C37), 115.0 (C33), 91.4 (C30), 74.6 (C11), 68.7 (C14), 59.5(Piperazinyl-C), 58.2 (C4), 52.5 (Piperazinyl-C), 45.5 (C22), 45.1 (C9), 44.0(C12,C13), 41.8 (C5), 36.7 (C6), 36.1 (C10), 34.4 (C2), 30.4 (C8), 26.8 (C7),26.4 (C18), 24.9 (C1), 20.9 (C38), 16.7 (C16), 14.9 (C15), 11.5 (C17). HRMS:calculated for C 38 H 50 N2O8([M + H] + ): 663.3640; found 663.3645.

[0065] Example 7: Synthesis of compound 5f was synthesized from compound 4, the specific synthesis steps were referred to Example 2.

[0066] Compound 5f white powder; yield: 69.2 %; melting point: 121.3-123.6 °C;

[0067] 1 H NMR (400 MHz, CDCl3): δ (ppm) 8.77 (d, J = 2.8 Hz, 1H, H36), 8.42(dd, J = 8.8, 2.8 Hz, 1H, H34), 7.45 (d, J = 8.8 Hz, 1H, H33), 6.49 (dd, J = 17.2,11.2 Hz, 1H, H19), 5.79 (d, J= 8.4 Hz, 1H, H14), 5.73 (s, 1H, H30), 5.34 (dd, J = 11.2, 1.6 Hz, 1H, H20), 5.20 (dd, J = 17.2, 1.6 Hz, 1H, H20), 4.96 (s, 2H,H28), 3.78 – 3.65 (m, 2H Piperazinyl-H), 3.60 – 3.47 (m, 2H Piperazinyl-H),3.39 – 3.32 (m, 1H, H11), 3.27 – 3.19 (m, 1H, H22), 3.15 – 3.08 (m, 1H, H22),2.73 – 2.57 (m, 4H, Piperazinyl-H)), 2.39 – 2.00 (m, 6H, H2, H4, H10, H13),1.84 – 1.33 (m, 9H, H1, H6, H7, H8, H15), 1.16 (s, 3H, H18), 1.15 – 1.08 (m,1H, H8), 0.88 (d, J = 6.8 Hz, 3H, H17), 0.70 (d, J = 6.8 Hz, 3H, H16). 13 C NMR (101MHz, CDCl3):δ (ppm) 217.0 (C3), 168.8 (C21), 163.7 (C29), 162.76 (C31), 156.8(C32), 143.9 (C35), 127.4 (C34), 119.8 (C36), 118.1 (C33), 117.3 (C20), 115.9(C37), 92.9 (C30), 74.6 (C11), 68.7 (C14), 59.5 (Piperazinyl-C), 58.2 (C4),52.4 (Piperazinyl-C), 45.5 (C22), 45.0 (C9), 44.0 (C12,C13), 41.8 (C5), 36.7(C6), 36.1 (C10), 34.5 (C2), 30.4 (C8), 26.8 (C7), 26.4 (C18), 24.9 (C1),16.7 (C16), 14.9 (C15), 11.5 (C17).HRMS: calculated for C37 H 47 N3O 10 ([M + H] + ):694.3334; found 694.3339.

[0068] Example 8: Synthesis of compound 5g was synthesized from compound 4, the specific synthesis steps refer to Example 2.

[0069] Compound 5g white powder; yield: 66.8 %; melting point: 115.6-118.9 °C;

[0070] 1 H NMR (400 MHz, CDCl3): δ (ppm) 7.87 (dd, J = 8.8, 6.0 Hz, 1H, H36),7.05 – 6.97 (m, 2H, H33, H35), 6.48 (dd, J = 17.2, 11.2 Hz, 1H, H19), 5.78 (d, J = 8.4 Hz, 1H, H14), 5.58 (s, 1H, H30), 5.34 (dd, J = 11.2, 1.2 Hz, 1H, H20),5.19 (dd, J = 17.2, 1.2 Hz, 1H, H20), 4.86 (s, 2H, H28), 3.76 – 3.66 (m,2HPiperazinyl-H), 3.57 – 3.45 (m, 2HPiperazinyl-H), 3.39 – 3.31 (m, 1H, H11),3.25 – 3.19 (m, 1H, H22), 3.14 – 3.045 (m, 1H, H22), 2.69 – 2.53 (m, 4H,Piperazinyl-H), 2.38 – 2.00 (m, 6H, H2, H4, H10, H13), 1.80 – 1.33 (m, 9H,H1, H6, H7, H8, H15), 1.16 (s, 3H, H18), 1.15 – 1.07 (m, 1H, H8), 0.88 (d, J =6.8 Hz, 3H, H17), 0.70 (d, J = 6.8 Hz, 3H, H16). 13C NMR (101 MHz, CDCl3): δ(ppm) 217.1 (C3), 168.8 (C21), 164.1 (C29), 163.2 (C34), 162.1 (C31), 149.5(C32), 139.1 (C19), 120.3 (C36), 117.3 (C20), 116.3 (C35), 109.1 (C37), 108.9(C33), 92.2 (C30), 74.6 (C11), 68.6 (C14), 59.5 (Piperazinyl-C), 58.2 (C4),52.4 (Piperazinyl-C), 45.5 (C22), 45.3 (C9), 44.9 (C13), 44.0 (C12), 41.9(C5), 36.7 (C6), 36.1 (C10), 34.5 (C2), 30.4 (C8), 26.8 (C7), 26.4 (C18),24.9 (C1), 16.7 (C16), 14.9 (C15), 11.5 (C17).HRMS: calculated for C 37 H 47 FN2O8([M + H] + ): 667.3389; found 667.3393.

[0071] Example 9: Synthesis of compound 5h was synthesized from compound 4, the specific synthesis steps refer to Example 2.

[0072] Compound 5h was a white powder; yield: 64.9 %; melting point: 112.4-114.6 °C;

[0073] 1 H NMR (400 MHz, CDCl3): δ (ppm) 7.53 (dd, J = 8.0, 2.4 Hz, 1H, H36),7.32 – 7.24 (m, 2H, H33, H34), 6.47 (dd, J = 17.2, 11.2 Hz, 1H, H19), 5.77 (d, J = 8.4 Hz, 1H, H14), 5.67 (s, 1H, H30), 5.32 (dd, J = 11.2, 1.2 Hz, 1H, H20),5.18 (dd,J = 17.2, 1.2 Hz, 1H, H20), 4.87 (s, 2H, H28), 3.76 – 3.61 (m,2HPiperazinyl-H), 3.59 – 3.44(m, 2H,Piperazinyl-H), 3.39 – 3.31 (m, 1H, H11),3.28 – 3.16 (m, 1H, H22), 3.15 – 3.04 (m, 1H, H22), 2.71 – 2.52 (m, 4H,Piperazinyl-H), 2.39 – 2.00 (m, 6H, H2, H4, H10, H13), 1.91 – 1.31 (m, 9H,H1, H6, H7, H8, H15), 1.15 (s, 3H, H18), 1.13 – 1.06 (m, 1H, H8), 0.87 (d, J =6.8 Hz, 3H, H17), 0.70 (d, J = 6.8 Hz, 3H, H16). 13 C NMR (101 MHz, CDCl3): δ(ppm) 217.0 (C3), 168.8 (C22), 164.1 (C29), 162.1 (C31), 157.5 (C35), 149.5(C32), 139.1 (C19), 120.3 (C33), 120.0 (C37), 118.4 (C34), 117.2 (20), 116.3(C36), 92.2 (C30), 74.6 (C11), 68.7 (C14), 59.5 (Piperazinyl-C), 58.2 (C4),52.4 (Piperazinyl-C), 45.5 (C22), 45.1 (C9), 44.0 (C12,C13), 41.8 (C5), 36.7(C6), 36.1 (C10), 34.4 (C2), 30.4 (C8), 26.8 (C7), 26.5 (C18), 24.9 (C1),16.7 (C16), 14.9 (C15), 11.5 (C17).HRMS: calculated for C 37 H 47 FN2O8([M + H] + ):667.3389; found 667.3389.

[0074] Example 10: Synthesis of compound 5i The synthesis of compound 5i was still synthesized from compound 4, the specific synthesis steps are referred to Example 2.

[0075] Compound 5i is a white powder; yield: 67.4 %; melting point: 109.4-112.6 °C;

[0076] 1 H NMR (400 MHz, CDCl3): δ (ppm)7.81 (d, J = 8.4 Hz, 1H, H36), 7.33 (d, J = 2.0 Hz, 1H, H35), 7.26 (dd, J = 8.4, 2.0 Hz, 1H, H33), 6.49 (dd, J = 17.2, 11.2Hz, 1H, H19), 5.79 (d, J = 8.4 Hz, 1H, H14), 5.62 (s, 1H, H30), 5.34 (dd, J =11.2, 1.2 Hz, 1H, H20), 5.20 (dd, J = 17.2, 1.2 Hz, 1H, H20), 4.86 (s, 2H,H28), 3.76 – 3.63 (m, 2H, Piperazinyl-H), 3.59 – 3.49 (m, 2H, Piperazinyl-H),3.40 – 3.32 (m, 1H, H11), 3.27 – 3.18 (m, 1H, H22), 3.15 – 3.06 (m, 1H, H22),2.73 – 2.53 (m, 4H, Piperazinyl-H), 2.40 – 1.99 (m, 6H, H2, H4, H10, H13),1.83 – 1.33 (m, 9H, H1, H6, H7, H8, H15), 1.16 (s, 3H, H18), 1.15 – 1.08 (m,1H, H8), 0.88 (d, J = 6.8 Hz, 3H, H17), 0.71 (d, J = 6.8 Hz, 3H, H16). 13C NMR (101 MHz, CDCl3): δ (ppm) 216.9 (C3), 168.8 (C21), 164.4 (C29), 163.2 (C31), 153.7(C32), 139.1 (C19), 138.7 (C34), 124.7 (C36), 124.2 (C35), 117.3 (C20), 117.1(C33), 114.0 (C37), 91.4 (C30), 74.6 (C11), 68.7 (C14), 59.5 (Piperazinyl-C),58.2 (C4), 52.4 (Piperazinyl-C), 45.5 (C22), 45.1 (C9), 44.0 (C12,C13), 41.8(C5), 36.7 (C6), 36.1 (C10), 34.4 (C2), 30.4 (C8), 26.9 (C7), 26.4 (C18),24.9 (C1), 16.729 (C16), 14.9 (C15), 11.5 (C17).HRMS: calculated forC 37 H 47 ClN2O8([M + H] + ): 683.3094; found 683.3099.

[0077] Example 11: Synthesis of compound 5j The synthesis of compound 5j was still synthesized from compound 4, the specific synthesis steps were referred to Example 2.

[0078] Compound 5j was a white powder; yield: 68.3 %; melting point: 108.9-111.2 °C;

[0079] 1 H NMR (400 MHz, CDCl3): δ (ppm) 7.85 (d, J = 2.4 Hz, 1H, H36), 7.51(dd, J = 8.8, 2.5 Hz, 1H, H34), 7.28 (d, J = 3.2 Hz, 1H, H33), 6.50 (dd, J = 17.2,11.2 Hz, 1H, H19), 5.80 (d, J = 8.4 Hz, 1H, H14), 5.67 (s, 1H, H30), 5.35 (dd,J = 11.2, 1.2 Hz, 1H, H20), 5.21 (dd, J = 17.2, 1.2 Hz, 1H, H20), 4.88 (s, 2H,H28), 3.78 – 3.66 (m, 2H, Piperazinyl-H), 3.61 – 3.50 (m, 2H, Piperazinyl-H),3.41 – 3.33 (m, 1H, H11), 3.28 – 3.19 (m, 1H, H22), 3.18 – 3.06 (m, 1H, H22),2.75 – 2.54 (m, 4H, Piperazinyl-H), 2.41 – 2.03 (m, 6H, H2, H4, H10, H13),1.84 – 1.34 (m, 9H, H1, H6, H7, H8, H15), 1.18 (s, 3H, H18), 1.16 – 1.09 (m,1H, H8), 0.90 (d, J = 6.8 Hz, 3H, H17), 0.72 (d, J = 6.8 Hz, 3H, H16). 13 C NMR (101MHz, CDCl3): δ (ppm) 216.9 (C3), 168.8 (C21), 163.2 (C31), 161.8 (C29), 151.8(C32), 139.1 (C19), 132.7 (C35), 129.7 (C34), 122.8 (C36), 118.3 (C37), 117.3(C20), 116.5 (C33), 92.2 (C30), 74.6 (C11), 68.7 (C14), 66.7 (s), 59.5(Piperazinyl-C), 58.2 (C14), 52.4 (Piperazinyl-C), 45.5 (C22), 45.1 (C9),44.0 (C12,C13), 41.8 (C5), 36.7 (C6), 36.1 (C10), 34.5 (C2), 30.4 (C8), 26.8(C7), 26.4 (C18), 24.9 (C1), 16.7 (C16), 14.9 (C15), 11.5 (C17).HRMS:calculated for C 37 H 47CIN2O8([M + H] + ): 683.3094; found 683.3097.

[0080] Example 12: Synthesis of compound 5k The synthesis of compound 5k was still synthesized from compound 4, the specific synthesis steps were referred to Example 2.

[0081] Compound 5k was a white powder; yield: 65.2 %; melting point: 121.3-124.4 °C;

[0082] 1 H NMR (400 MHz, CDCl3): δ (ppm) 7.73 (d, J = 8.4 Hz, 1H, H33), 7.50 (d, J = 1.6 Hz, 1H, H36), 7.41 (dd, J = 8.4, 1H, H35), 6.49 (dd, J = 17.2, 11.2 Hz, 1H, H19), 5.79 (d, J = 8.4 Hz, 1H, H14), 5.63 (s, 1H, H30), 5.34 (dd, J = 11.2, 1.2 Hz, 1H, H20), 5.20 (dd, J = 17.2, 1.2 Hz, 1H, H20), 4.85 (s, 2H,H28), 3.77 – 3.63 (m, 2H, Piperazinyl-H), 3.56 – 3.47 (m, 2H, Piperazinyl-H),3.41 – 3.31 (m, 1H, H11), 3.26 – 3.18 (m, 1H, H22), 3.16 – 3.04 (m, 1H, H22),2.74 – 2.53 (m, 4H, Piperazinyl-H), 2.41 – 2.00 (m, 6H, H2, H4, H10, H13),1.85 – 1.32 (m, 9H, H1, H6, H7, H8, H15), 1.16 (s, 3H, H18), 1.15 – 1.08 (m,1H, H8), 0.88 (d, J = 6.8 Hz, 3H, H17), 0.71 (d, J= 6.8 Hz, 3H, H16). 13 C NMR (101 MHz, CDCl3): δ (ppm) 216.9 (C3), 168.8 (C21), 164.4 (C29), 163.2 (C31), 153.6(C32), 139.1 (C19), 127.5 (C36), 126.7 (C35), 124.3 (C34), 120.1 (C33), 117.3(C20), 114.4 (C37), 91.6 (C30), 74.6 (C14), 68.7 (C14), 59.5 (Piperazinyl-C),58.2 (C4), 52.4 (Piperazinyl-C), 45.5 (C22), 45.1 (C9), 44.8 (C13), 44.0(C12), 41.8 (C5), 36.7 (C6), 36.1 (C10), 34.4 (C2), 30.4 (C8), 26.9 (C7),26.4 (C18), 24.9 (C1), 16.7 (C16), 14.9 (C15), 11.5 (C17).HRMS: calculated for C 37 H 47 BrN2O8([M + H] + ): 727.2589; found 727.2584.

[0083] Example 13: Synthesis of compound 5l The synthesis of compound 5l was still synthesized from compound 4, the specific synthesis steps were referred to Example 2.

[0084] Compound 5l was a white powder; yield: 64.5 %; melting point: 110.8-112.6 °C;

[0085] 1 H NMR (400 MHz, CDCl3): δ (ppm) 7.98 (d, J = 2.4 Hz, 1H, H36), 7.63(dd, J = 8.8, 2.4 Hz, 1H, H34), 7.19 (d, J = 8.8 Hz, 1H, H33), 6.48 (dd, J = 17.2,11.2 Hz, 1H, H19), 5.78 (d, J= 8.4 Hz, 1H, H14), 5.65 (s, 1H, H30), 5.33 (dd, J = 11.2, 1.2 Hz, 1H, H20), 5.19 (dd, J = 17.2, 1.2 Hz, 1H, H20), 4.87 (s, 2H,H28), 3.76 – 3.62 (m, 2H, Piperazinyl-H), 3.57 – 3.46 (m, 2H, Piperazinyl-H),3.39 – 3.30 (m, 1H, H11), 3.26 – 3.15 (m, 1H, H22), 3.14 – 3.05 (m, 1H, H22),2.74 – 2.52 (m, 4H, Piperazinyl-H), 2.40 – 2.01 (m, 6H, H2, H4, H10, H13),1.86 – 1.32 (m, 9H, H1, H6, H7, H8, H15), 1.15 (s, 3H, H18), 1.14 – 1.07 (m,1H, H8), 0.87 (d, J = 6.8 Hz, 3H, H17), 0.70 (d, J = 6.8 Hz, 3H, H16). 13C NMR (101 MHz, CDCl3): δ (ppm) 216.9 (C3), 168.8 (C21), 163.7 (C29), 163.2 (C31), 161.7(C32), 152.2 (C34), 139.1 (C19), 135.5 (C36), 125.8 (C35), 118.5 (C37), 117.3(C20), 116.9 (C33), 92.2 (C30), 74.6 (C11), 68.7 (C14), 59.5 (Piperazinyl-C),58.2 (C4), 52.4 (Piperazinyl-C), 45.5 (C22), 45.1 (C9), 44.0 (C12,C13), 41.8(C5), 36.7 (C6), 36.1 (C10), 34.4 (C2), 30.4 (C8), 26.8 (C7), 26.4 (C18),24.9 (C1), 16.7 (C16), 14.9 (C15), 11.5 (C17).HRMS: calculated for C 37 H 47 BrN2O8([M + H] + ): 727.2589; found 727.2587.

[0086] Example 14: Synthesis of compound 5m The synthesis of compound 5m was still synthesized from compound 4, the specific synthesis steps were referred to Example 2.

[0087] Compound 5m was a white powder; yield: 56.8 %; melting point: 106.5-108.4 °C;

[0088] 1 H NMR (400 MHz, CDCl3): δ (ppm) 7.94 (s, 1H, H36), 7.47 (d, J = 8.8 Hz,1H, H34), 7.23 (d, J = 8.8 Hz, 1H, H33), 6.69 (s, 1H, -NH), 6.49 (dd, J = 17.2,11.2 Hz, 1H, H19), 5.78 (d, J = 8.4 Hz, 1H, H14), 5.62 (s, 1H, H30), 5.33 (d, J= 11.2 Hz, 1H, H20), 5.19 (d, J = 17.2 Hz, 1H, H20), 4.84 (s, 2H, H28), 3.75 –3.64 (m, 2H, Piperazinyl-H), 3.60 – 3.47 (m, 2H, Piperazinyl-H), 3.39 – 3.32(m, 1H, H11), 3.26 – 3.17 (m, 1H, H22), 3.14 – 3.06 (m, 1H, H22), 2.73 – 2.52(m, 4H, Piperazinyl-H), 2.38 – 2.03 (m, 6H, H2, H4, H10, H13), 1.83 – 1.32(m, 18H, H1, H6, H7, H8, H15, Boc-H), 1.16 (s, 3H, H18), 1.14 – 1.07 (m, 1H,H8), 0.88 (d, J = 6.8 Hz, 3H, H17), 0.70 (d, J = 6.8 Hz, 3H, H16). 13 C NMR (101MHz, CDCl3): δ (ppm) 217.0 (C3), 168.8 (C21), 164.6 (C29), 163.5 (C31), 152.4(C38),149.1 (C32), 139.1 (C19), 134.9 (C35), 123.3 (C34,C37), 117.3 (C20),115.5 (C33,C36), 91.8 (C30), 80.9 (C39), 68.60 (C14), 59.6 (Piperazinyl-C),58.2 (C4), 52.8 (Piperazinyl-C), 45.5 (C22), 45.1 (C9), 44.0 (C12,C13), 41.8(C5), 36.7 (C6), 36.1 (C10), 34.5 (C2), 30.4 (C8), 28.3 (C40,C41,C42), 26.8(C7), 26.4 (C18), 24.9 (C1), 16.7 (C16), 14.9 (C15), 11.5 (C17).HRMS:calculated for C 42 H 57 N3O10 ([M + H] + ): 764.4117; found 764.4115.

[0089] Example 15: Synthesis of compound 6a The synthesis of compound 6a was still synthesized from compound 4, the specific synthesis steps were referred to Example 2.

[0090] Compound 6a was a white powder; yield: 76.9 %; melting point: 94.5-97.2 °C;

[0091] 1 H NMR (400 MHz, CDCl3): δ (ppm) 7.66 (d, J = 9.6 Hz, 1H, H36), 7.32 –7.23 (m, 1H, H32), 7.15 (dd, J = 9.2, 2.8 Hz, 1H, H37), 7.02 (d, J = 2.8 Hz, 1H,H30), 6.62 – 6.35 (m, 2H, H19, H33), 5.78 (d, J = 8.4 Hz, 1H, H14), 5.37 – 5.28(m, 1H, H20), 5.19 (dd, J = 17.2, 1.2 Hz, 1H, H20), 4.74 (s, 2H, H28), 3.80 –3.65 (m, 2H, Piperazinyl-H), 3.65 – 3.56 (m, 2H, Piperazinyl-H), 3.40 – 3.33(m, 1H, H11), 3.27 – 3.15 (m, 1H, H22), 3.14 – 3.00 (m, 1H, H22), 2.76 – 2.46(m, 4H, Piperazinyl-H), 2.41 – 1.91 (m, 6H, H2,H4, H10, H13), 1.85 – 1.33(m, 9H, H1, H6, H7, H8, H15), 1.16 (s, 3H, H18), 1.15 – 1.07 (m, 1H, H8),0.89 (d, J = 6.8 Hz, 3H, H17), 0.71 (d, J = 6.8 Hz, 3H, H16). 13C NMR (101 MHz, CDCl3) δ 217.0 (C3), 168.8 (C21), 165.9(C29), 160.8 (C32), 154.4(C31), 143.1(C34), 139.2 (C19), 119.9(C36), 119.3(C20), 118.1 (C33), 117.2 (C35), 117.1(C37), 111.5(C30), 74.6 (C11), 68.6(C14), 59.5(Piperazinyl-C), 58.2(Piperazinyl-C), 52.8(Piperazinyl-C), 52.3(Piperazinyl-C), 45.4 (C22), 45.1(C9), 44.0, (C13) 42.0(C5), 36.7(C6), 36.1(C19), 34.4(C2), 30.4(C8), 26.8(C7), 26.5 (C18), 24.8(C1), 16.7 (C16), 14.9 (C15), 11.5. (C17). HRMS:calculated for C 37 H 48 N₂O₈([M + H]) + ): 649.3483; found 649.3494.

[0092] Example 16: Compound 7a The synthesis of [the compound] is still performed using compound 4, and the specific synthesis steps are described in Example 2.

[0093] Compound 7a is a white powder; yield: 69.9%; melting point: 96.4-98.4℃;

[0094] 1 H NMR (400 MHz, CDCl3): δ (ppm) 7.62 (d, J = 9.6 Hz, 1H, H34), 7.37 (d, J = 8.4 Hz, 1H, H36), 6.89 (dd, J = 8.4, 2.4 Hz, 1H, H30), 6.80 (d, J = 2.4 Hz, 1H, H37), 6.46 (dd, J = 17.2, 11.2 Hz, 1H, H19), 6.24 (d, J= 9.6 Hz, 1H, H33), 5.76 (d, J = 8.4 Hz, 1H, H14), 5.31 (dd, J = 11.0, 1.2 Hz, 1H, H20), 5.18 (dd, J = 17.2, 1.2 Hz, 1H, H20), 4.74 (s, 2H, H28), 3.79 - 3.62 (m, 2H, Piperazinyl-H), 3.61 - 3.50 (m, 2H, Piperazinyl-H), 3.43 - 3.29 (m, 1H, H11), 3.27 - 3.14 (m, 1H, H22), 3.14 - 3.00 (m, 1H, H22), 2.71 - 2.45 (m, 4H, Piperazinyl-H), 2.40 - 2.01 (m, 6H, H2, H4, H10, H13), 1.81 - 1.31 (m, 9H, H1, H6, H7, H8, H15), 1.14 (s, 3H, H18), 1.13 - 1.06 (m, 1H, H8), 0.86 (d, J = 6.8 Hz, 3H, H17), 0.69 (d, J = 6.8 Hz, 3H, H16). 13C NMR (101 MHz, CDCl3) δ (ppm) 217.0 (C3),168.8 (C21), 165.2 (C29), 160.9 (C32), 155.7 (C31), 143.2 (C34), 139.1 (C19),129.0 (C36), 117.2 (C20), 113.7 (C33), 113.3 (C35), 112.7 (C37), 102.0 (C30),74.6 (C11), 68.6 (C14), 59.6 (Piperazinyl-C), 58.2 (Piperazinyl-C), 52.8(Piperazinyl-C), 52.3 (Piperazinyl-C), 45.5 (C22), 45.1 (C9), 44.0 (C13),41.9 (C5), 36.7 (C6), 36.1 (C19), 34.4 (C2), 30.4 (C8), 26.8 (C7), 26.5(C18), 24.9 (C1), 16.7 (C16), 14.9 (C15), 11.5 (C17).HRMS: calculated forC 37 H 48 N2O8([M + H] + ): 649.3483; found 649.3488.

[0095] Example 17: Synthesis of compound 8a was synthesized from compound 4, the specific synthesis steps are referred to Example 2.

[0096] Compound 8a is a white powder; yield: 56 %; melting point: 96.4-98.4 °C;

[0097] 1 H NMR (400 MHz, CDCl3): δ (ppm) 8.83 (s, 1H, H36), 8.06 (d, J = 8.0Hz,1H, H34), 7.36 (s, 3H, H31, H32, H35), 7.12 (d, J = 3.6 Hz, 1H, H30), 6.37 (dd, J = 17.2, 11.2 Hz, 1H, H19), 5.67 (d, J = 8.0 Hz, 1H, H14), 5.22 (d,J = 10.8 Hz,1H, H20), 5.09 (d, J = 17.2 Hz, 1H, H20), 4.94 (s, 2H, H28), 3.78 – 3.66 (m,2H, Piperazinyl-H), 3.64 – 3.47 (m, 2H, Piperazinyl-H), 3.33 – 3.22 (m, 1H,H11), 3.07 – 2.97 (m, 1H, H22), 2.96 – 2.83 (m, 1H, H22), 2.50 – 2.33 (m,4H, Piperazinyl-H), 2.31 – 1.86 (m, 6H, H2,H4, H10, H13), 1.82 – 1.21 (m,9H, H1, H6, H7, H8, H15), 1.07 (s, 3H, H18), 1.04 – 0.95 (m, 1H, H8), 0.80(d, J = 6.4Hz, 3H), 0.58 (d, J = 6.4 Hz, 3H, H16). 13 C NMR (101 MHz, CDCl3): δ(ppm) 217.0 (C3), 168.8 (C21), 166.4 (C29), 153.4 (C36), 149.3 (C37), 139.1(C19), 136.0 (C34), 129.5 (C33), 126.64 (C31), 121.7 (C35), 120.8 (C32),117.0 (C20), 110.1 (C30), 74.5 (C11), 59.6 (Piperazinyl-C), 58.1(Piperazinyl-C), 52.9 (Piperazinyl-C), 52.4 (Piperazinyl-C), 45.4 (C22), 44.7(C9), 43.9 (C13), 42.0 (C12), 41.7 (C5), 36.6 (C6), 36.0 (C19), 34.4 (C2),30.4 (C8), 26.8 (C7), 26.7 (C18), 24.8 (C1), 16.6 (C16), 14.8 (C15), 11.4(C17). IR (KBr, cm -1) : 3436, 1731, 1633, 1574, 1504, 1470, 1454, 1384, 1120.HRMS: calculated for C 37 H 49 N3O6([M + H] + ): 632.3694; found 632.3692.

[0098] Example 18: Synthesis of compound 8b The synthesis of compound 8b was still synthesized from compound 4, the specific synthesis steps were referred to Example 2.

[0099] Compound 8b was a white powder; yield: 54 %; melting point: 101.2-104.3 °C;

[0100] 1 H NMR (400 MHz, CDCl3): δ (ppm)7.94 (d, J = 8.4 Hz, 1H, H34), 7.39 –7.27 (m, 2H), 7.23 (d, J = 8.4 Hz, 1H), 7.05 (d, J = 7.2 Hz, 1H, H30), 6.39 (dd, J = 17.2, 11.2 Hz, 1H, H19), 5.69 (d, J = 8.4 Hz, 1H, H14), 5.24 (d, J = 11.2 Hz,1H, H20), 5.11 (d, J = 17.2 Hz, 1H, H20), 4.92 (s, 2H, H28), 3.75 (s, 2H,Piperazinyl-H), 3.61 (s, 2H, Piperazinyl-H), 3.29 (d, J= 4.8 Hz, 1H, H11),3.10 – 3.03 (m, 1H, H22) , 2.98 – 2.90 (m, 1H, H22), 2.68 (s, 3H, Me-H), 2.55– 2.37 (m, 4H, Piperazinyl-H), 2.32 – 1.89 (m, 6H, H2 ,H4, H10, H13), 1.77 –1.24 (m, 9H, H1, H6, H7, H8, H15), 1.08 (s, 3H, H18), 1.06 – 0.98 (m, 1H,H8), 0.81 (d, J = 6.8 Hz, 3H, H17), 0.61 (d, J = 6.8 Hz, 3H, H16). 13 C NMR (101MHz, CDCl3): δ (ppm) 217.0 (C3), 168.8 (C21), 158.1 (C36), 152.9 (C29), 139.7(C37), 139.2 (C19), 136.1 (C34), 127.7 (C33), 125.6 (C31), 122.6 (C35), 120.8(C32), 117.0 (C20), 110.5 (C30), 74.5 (C11), 68.6 (C14), 59.6 (Piperazinyl-C), 58.1 (Piperazinyl-C), 53.0 (Piperazinyl-C), 52.5 (Piperazinyl-C), 45.4(C22), 45.1 (C9), 43.9 (C13), 41.8 (C12), 41.7 (C5), 36.6 (C6), 36.1 (C19),34.4 (C2), 30.4 (C8), 26.8 (C7), 26.6 (C18), 25.4 (C38), 24.8 (C1), 16.6(C16), 14.8 (C15), 11.4 (C17). IR (KBr, cm -1 ) : 3444, 2932, 1732, 1651, 1505,1455, 1434, 1382, 1260, 1235, 1191, 1153, 1118, 1010. HRMS: calculated forC 38 H 51N3O6([M + H] + ): 646.3851; found 646.3853.

[0101] Example 19: Synthesis of compound 8c The synthesis of compound 8c was still synthesized from compound 4, the specific synthesis steps were referred to Example 2.

[0102] Compound 8c was a white powder; yield: 54 %; melting point: 97.3-99.6 °C;

[0103] 1 H NMR (400 MHz, CDCl3): δ (ppm)8.70 (d, J = 2.0 Hz, 1H, H36), 7.85 (s,1H, H34), 7.41 – 7.28 (m, 2H, H32, H31), 7.12 – 7.04 (m, 1H, H30), 6.41 (dd, J = 17.2, 11.2 Hz, 1H, H19), 5.70 (d, J = 8.4 Hz, 1H, H14), 5.26 (dd, J = 9.6,1.6 Hz, 1H, H20), 5.13 (dd, J = 17.2, 1.2 Hz, 1H, H20), 4.96 (s, 2H, H28), 3.83– 3.70 (m, 2H, Piperazinyl-H), 3.65 – 3.54 (m, 2H, Piperazinyl-H), 3.30 (d, J =4.8 Hz, 1H, H11), 3.11 – 2.99 (m, 1H, H22), 3.65 – 3.53 (m, 1H, H22), 2.47(s, 3H, Me-H), 2.31 – 1.92 (m, 6H, H2,H4, H10, H13), 1.79 – 1.25 (m, 9H, H1,H6, H7, H8, H15), 1.10 (s, 3H, H18), 1.18 – 1.00 (m, 1H, H8), 0.83 (d, J = 6.8Hz, 3H, H17), 0.62 (d, J = 6.8 Hz, 3H, H16). 13C NMR (101 MHz, CDCl3): δ (ppm) 217.0 (C3), 168.8 (C21), 153.5 (C29), 151.2 (C36), 139.1 (C19), 138.3 (C37), 134.7 (C34), 131.2 (C35), 129.4 (C33), 126.7 (C31), 120.2 (C32), 117.1 (C20), 109.1 (C30), 74.5 (C11), 68.5 (C14), 59.6 (Piperazinyl-C), 58.1 (Piperazinyl-C), 53.0 (Piperazinyl-C), 52.5 (Piperazinyl-C), 45.5 (C22), 44.8 (C9), 43.9 (C13), 42.0 (C12), 41.7 (C5), 36.7 (C6), 36.1 (C19), 34.4 (C2), 30.4 (C8), 26.8 (C7), 26.5 (C18), 24.8 (C1), 18.6 (C38), 16.6 (C16), 14.8 (C15), 11.4 (C17). IR (KBr, cm -1 ) : 3417, 1731, 1659, 1651, 1643, 1633, 1454, 1402, 1262, 1107. HRMS: calculated for C 38 H 51 N3O6([M + H] + ): 646.3851; found 646.3851.

[0104] Example 20: Synthesis of compound 8d was synthesized from compound 4, the specific synthesis steps were referred to Example 2.

[0105] Compound 8d was a white powder; yield: 53 %; melting point: 94.6-98.4 °C;

[0106] 1 H NMR (400 MHz, CDCl3): δ (ppm) 8.93 – 8.79 (m, 1H, H36), 8.23 (dd, J =8.4, 1.4 Hz, 1H, H34), 7.46 – 7.35 (m, 1H, H35), 7.20 (d,J = 8.0 Hz, 1H,H31), 7.03 (d, J = 8.0 Hz, 1H, H30), 6.40 (dd, J = 17.2, 11.2 Hz, 1H, H19), 5.70(d, J = 8.4 Hz, 1H, H14), 5.25 (d, J = 11.2 Hz, 1H, H20), 5.12 (d, J = 17.2 Hz, 1H,H20), 4.94 (s, 2H, H28), 3.80 – 3.67 (m, 2H, Piperazinyl-H), 3.67 – 3.53 (m,2H, Piperazinyl-H), 3.36 – 3.24 (m, 1H, H11), 3.11 – 3.00 (m, 1H, H22), 2.99 J = 6.8 Hz, 3H,H17), 0.61 (d, J = 6.8 Hz, 3H, H16). 13C NMR (101 MHz, CDCl3): δ (ppm) 217.0(C3), 168.8 (C21), 166.7 (C29), 152.0 (C35), 148.8 (C37), 139.1 (C19), 132.6(C34), 128.6 (C33), 127.2 (C32), 126.6 (C31), 121.3 (C35), 117.1 (C20), 109.8(C30), 74.5 (C11), 68.5 (C14), 59.6 (Piperazinyl-C), 58.1 (Piperazinyl-C),52.9 (Piperazinyl-C), 52.5 (Piperazinyl-C), 45.5 (C22), 44.8 (C9), 43.9(C13), 42.0 (C12), 41.7 (C5), 36.7 (C6), 36.1 (C19), 34.4 (C2), 30.4 (C8),26.8 (C7), 26.5 (C18), 24.8 (C1), 18.1 (C38), 16.6 (C16), 14.8 (C15), 11.4(C17). IR (KBr, cm -1 ) : 3443, 2933, 1731, 1657, 1642, 1401, 1384, 1263, 1246,1194, 1152, 1117, 1010. HRMS: calculated for C 38 H 51 N3O6([M + H] + ): 646.3851;found 646.3849.

[0107] Example 21: Synthesis of compound 8e was synthesized from compound 4, the specific synthesis steps are referred to Example 2.

[0108] Compound 8e is a white powder; yield: 68 %; melting point: 97.4-101.5 °C;

[0109] 1 H NMR (400 MHz, CDCl3): δ (ppm)9.12 (d, J = 8.8 Hz, 1H, H34), 9.01 –8.87 (m, 1H, H36), 8.39 (d, J= 8.8 Hz, 1H, H31), 7.63 (dd, J = 8.8, 4.0 Hz, 1H,H35), 7.13 (d, J = 8.8 Hz, 1H, H30), 6.38 (dd, J = 17.2, 11.2 Hz, 1H, H19), 5.68(d, J = 8.4 Hz, 1H, H14), 5.23 (d, J = 10.8 Hz, 1H, H20), 5.18 – 5.03 (m, 3H,H20, H28), 3.70 – 3.63 (m, 2H, Piperazinyl-H), 3.62 – 3.53 (m, 2H,Piperazinyl-H), 3.29 (d, J = 6.0 Hz, 1H, H11), 3.14 – 3.04 (m, 1H, H22), 3.03 –2.92 (m, 1H, H22), 2.56 – 2.41 (m, 4H, Piperazinyl-H), 2.28 – 1.97 (m, 6H, H2,H4, H10, H13), 1.79 – 1.25 (m, 9H, H1, H6, H7, H8, H15), 1.08 (s, 3H, H18),1.06 – 0.99 (m, 1H, H8), 0.82 (d, J = 6.8 Hz, 3H, H17), 0.60 (d, J = 6.8 Hz, 3H,H16). 13C NMR (101 MHz, CDCl3): δ (ppm) 216.9 (C3), 168.7 (C21), 164.9 (C29),158.9 (C36), 150.2 (C32), 139.2 (C19), 138.4 (C37), 132.5 (C34), 127.1 (C31),124.6 (C35), 123.0 (C33), 117.0 (C20), 107.4 (C30), 74.5 (C11), 68.5 (C14),59.4 (Piperazinyl-C), 58.1 (Piperazinyl-C), 52.7 (Piperazinyl-C), 52.2(Piperazinyl-C), 45.4 (C22), 44.7 (C9), 43.9 (C13), 42.1 (C12), 41.7 (C5),36.6 (C6), 36.0 (C19), 34.4 (C2), 30.4 (C8), 26.8 (C7), 26.5 (C18), 24.8(C1), 16.6 (C16), 14.8 (C15), 11.4 (C17). IR (KBr, cm -1 ) : 3455, 1659, 1642,1632, 1513, 1503, 1462, 1452, 1402, 1240, 1118, 1016, 941, 906, 831. HRMS:calculated for C 37 H 48 N4O8([M + H] + ): 677.3545; found 677.3550.

[0110] Example 22: Synthesis of compound 8f was synthesized from compound 4, the specific synthesis steps were referred to Example 2.

[0111] Compound 8f was a white powder; yield: 58 %; melting point: 95.4-98.7 °C;

[0112] 1 H NMR (400 MHz, CDCl3): δ (ppm)8.86 (dd, J = 4.0, 1.2 Hz, 1H, H36),8.44 (d, J= 8.4 Hz, 1H, H34), 7.50 – 7.37 (m, 2H, H31, H35), 7.04 (d, J = 8.4 Hz, 1H, H30), 6.36 (dd, J = 17.2, 11.2 Hz, 1H, H19), 5.66 (d, J = 8.4 Hz, 1H,H14), 5.21 (d, J = 11.2 Hz, 1H, H20), 5.08 (d, J = 17.2 Hz, 1H, H20), 4.92 (s,2H, H28), 3.75 – 3.62 (m, 2H, Piperazinyl-H), 3.61 – 3.49 (m, 2H,Piperazinyl-H), 3.27 (d, J = 6.0 Hz, 1H, H11), 3.11 – 2.99 (m, 1H, H22), 2.98 –2.85 (m, 1H, H22), 2.50 – 2.33 (m, 4H, Piperazinyl-H), 2.27 – 1.92 (m, 6H, H2,H4, H10, H13), 1.73 – 1.20 (m, 9H, H1, H6, H7, H8, H15), 1.06 (s, 3H, H18),1.03 – 0.97 (m, 1H, H8), 0.79 (d, J = 6.8 Hz, 3H, H17), 0.58 (d, J = 6.8 Hz, 3H,H16). 13C NMR (101 MHz, CDCl3): δ (ppm) 216.9 (C3), 168.7 (C21), 166.0 (C36),152.7 (C29), 149.7 (C37), 139.1 (C19), 133.0 (C34), 127.1 (C31), 126.4 (C33),123.3 (C35), 122.4 (C32), 117.0 (C20), 110.0 (C30), 74.5 (C11), 68.5 (C14),59.5 (Piperazinyl-C), 58.1 (Piperazinyl-C), 52.8 (Piperazinyl-C), 52.3(Piperazinyl-C), 45.4 (C22), 45.4 (C9), 43.9 (C13), 41.8 (C12), 41.7 (C5),36.6 (C6), 36.0 (C19), 34.4 (C2), 30.3 (C8), 26.8 (C7), 26.6 (C18), 24.8(C1), 16.6 (C16), 14.8 (C15), 11.4 (C17). IR (KBr, cm -1 ) : 3439, 2924, 1731,1644, 1504, 1384, 1309, 1102, 789. HRMS: calculated for C 37 H 48 ClN3O6([M + H] + ):666.3304; found 666.3304.

[0113] Example 23: Synthesis of compound 8g was synthesized from compound 4, the specific synthesis steps were referred to Example 2.

[0114] Compound 8g was a white powder; yield: 63 %; melting point: 101.2-103.6 °C;

[0115] 1 H NMR (400 MHz, CDCl3): δ (ppm)7.62 (d, J = 9.6 Hz, 1H, H34), 7.37 (d, J = 8.4 Hz, 1H, H36), 6.89 (dd, J= 8.4, 2.4 Hz, 1H, H30), 6.80 (d, J = 2.4 Hz,1H, H37), 6.46 (dd, J = 17.2, 11.2 Hz, 1H, H19), 6.24 (d, J = 9.6 Hz, 1H, H33),5.76 (d, J = 8.4 Hz, 1H, H14), 5.31 (dd, J = 11.0, 1.2 Hz, 1H, H20), 5.18 (dd, J =17.2, 1.2 Hz, 1H, H20), 4.74 (s, 2H, H28), 3.79 – 3.62 (m, 2H, Piperazinyl-H), 3.61 – 3.50 (m, 2H, Piperazinyl-H), 3.43 – 3.29 (m, 1H, H11), 3.27 – 3.14(m, 1H, H22), 3.14 – 3.00 (m, 1H, H22), 2.71 – 2.45 (m, 4H, Piperazinyl-H),2.40 – 2.01 (m, 6H, H2,H4, H10, H13), 1.81 – 1.31 (m, 9H, H1, H6, H7, H8,H15), 1.14 (s, 3H, H18), 1.13 – 1.06 (m, 1H, H8), 0.86 (d, J = 6.8 Hz, 3H,H17), 0.69 (d, J = 6.8 Hz, 3H, H16). 13C NMR (101 MHz, CDCl3): δ (ppm) 217.0(C3), 168.8 (C21), 165.2 (C29), 160.9 (C32), 155.7 (C31), 143.2 (C34), 139.1(C19), 129.0 (C36), 117.2 (C20), 113.7 (C33), 113.3 (C35), 112.7 (C37), 102.0(C30), 74.6 (C11), 68.6 (C14), 59.6 (Piperazinyl-C), 58.2 (Piperazinyl-C),52.8 (Piperazinyl-C), 52.3 (Piperazinyl-C), 45.5 (C22), 45.1 (C9), 44.0(C13), 41.9 (C5), 36.7 (C6), 36.1 (C19), 34.4 (C2), 30.4 (C8), 26.8 (C7),26.5 (C18), 24.9 (C1), 16.7 (C16), 14.9 (C15), 11.5 (C17). IR (KBr, cm -1 ) :3444, 2928, 1731, 1644, 1486, 1469, 1499, 1384, 1293, 1243, 1203, 1153, 1116,1010. HRMS: calculated for C 37 H 47 Cl2N3O6([M + H] + ): 700.2915; found 700.2912.

[0116] Example 24: Synthesis of compound 8h was synthesized from compound 4, the specific synthesis steps were referred to Example 2.

[0117] Compound 8h was a white powder; yield: 56 %; melting point: 104.4-108.2 °C;

[0118] 1H NMR (400 MHz, CDC13): δ (ppm) 8.91 - 8.84 (m, 1H, H36), 8.52 - 8.38 (m, 1H, H34), 7.71 - 7.60 (m, 1H, H31), 7.54 - 7.45 (m, 1H, H35), 7.10 - 6.99 (m, 1H, H30), 6.41 (dd, J = 17.2, 11.2, 1H, H19), 5.76 - 5.63 (m, 1H, H14), 5.26 (dd, J = 10.8, 2.8 Hz, 1H, H20), 5.18 - 5.08 (m, 1H, H20), 5.02 - 4.90 (m, 2H, H28), 3.81 - 3.69 (m, 2H, Piperazinyl-H), 3.64 - 3.54 (m, 2H, Piperazinyl-H), 3.35 - 3.25 (m, 1H, H11), 3.13 - 3.02 (m, 1H, H22), 3.01 - 2.89 (m, 1H, H22), 2.52 - 2.35 (m, 4H, Piperazinyl-H), 2.33 - 1.92 (m, 6H, H2, H4, H10, H13), 1.78 - 1.26 (m, 9H, H1, H6, H7, H8, H15), 1.10 (d, J = 2.4 Hz, 3H, H18), 1.08 - 1.00 (m, 1H, H8), 0.89 - 0.77 (m, 3H, H17), 0.61 (dd, J = 6.8, 2.5 Hz, 3H, H16). 13C NMR (101 MHz, CDCl3): δ (ppm) 217.0 (C3), 168.8 (C21),153.4 (C29), 149.8 (C36), 140.8 (C37), 139.1 (C19), 135.6 (C34), 130.1 (C31),128.4 (C33), 122.8 (C35), 117.2 (C20), 113.2 (C30), 110.7 (C32), 74.5 (C11),68.5 (C14), 59.6 (Piperazinyl-C), 58.1 (Piperazinyl-C), 52.8 (Piperazinyl-C),52.4 (Piperazinyl-C), 45.4 (C22), 45.1 (C9), 43.9 (C13), 41.9 (C12), 41.7(C5), 36.6 (C6), 36.0 (C19), 34.4 (C2), 30.4 (C8), 26.8 (C7), 26.5 (C18),24.8 (C1), 16.6 (C16), 14.8 (C15), 11.5 (C17). IR (KBr, cm -1 ) : 3422, 2928,1733, 1647, 1498, 1458, 1401, 1383, 1362, 1307, 1228, 1193, 1162, 1116, 1101,1010. HRMS: calculated for C 37 H 48 BrN3O 6 ([M + H] + ): 710.2799; found 710.2797.

[0119] Example 25: Synthesis of compound 8i was synthesized from compound 4, the specific synthesis steps were referred to Example 2.

[0120] Compound 8i was a white powder; yield: 56 %; melting point: 107.4-110.6 °C;

[0121] 1 H NMR (400 MHz, CDCl3): δ (ppm)8.86 (dd, J = 4.0, 1.6 Hz, 1H, H36),8.44 (dd, J= 8.4, 1.6 Hz, 1H, H34), 7.95 (s, 1H, H35), 7.50 (dd, J = 8.4, 4.0Hz, 1H, H31), 6.47 (dd, J = 17.2, 11.2 Hz, 1H, H19), 5.76 (d, J = 8.4 Hz, 1H,H14), 5.30 (d, J = 11.7 Hz, 1H, H20), 5.16 (dd, J = 17.2, 1.2 Hz, 1H, H20), 5.10(s, 2H, H28), 4.00 – 3.83 (m, 2H, Piperazinyl-H), 3.80 – 3.63 (m, 2H,Piperazinyl-H), 3.33 (d, J = 5.6 Hz, 1H, H11), 3.26 – 3.15 (m, 1H, H22), 3.12 –3.03 (m, 1H, H22), 2.76 – 2.49 (m, 4H, Piperazinyl-H), 2.37 – 1.99 (m, 6H, H2,H4, H10, H13), 1.80 – 1.30 (m, 9H, H1, H6, H7, H8, H15), 1.13 (s, 3H, H18),1.11 – 1.03 (m, 1H, H8), 0.85 (d, J = 6.8 Hz, 3H, H17), 0.69 (d, J = 6.8 Hz, 3H,H16). 13C NMR (101 MHz, CDCl3): δ (ppm) 217.0 (C3), 168.9 (C21), 153.4 (C36),149.8 (C29), 140.8 (C37), 139.2 (C19), 135.6 (C34), 130.1 (C31), 128.4 (C33),122.8 (C35), 117.2 (C20), 113.2 (C30), 110.7 (C32), 74.6 (C11), 68.5 (C14),59.7 (Piperazinyl-C), 58.2 (Piperazinyl-C), 53.1 (Piperazinyl-C), 52.5(Piperazinyl-C), 45.4 (C22), 45.0 (C9), 43.9 (C13), 42.0 (C12), 41.7 (C5),36.7 (C6), 36.1 (C19), 34.4 (C2), 30.4 (C8), 26.8 (C7), 26.5 (C18), 24.8(C1), 16.7 (C16), 14.9 (C15), 11.5 (C17). IR (KBr, cm -1 ) : 3439, 2928, 1732,1651, 1644, 1574, 1454, 1384, 1345, 1262, 1202, 1116, 1089, 1011, 923, 859,799. HRMS: calculated for C 37 H 47 Br2N3O6([M + H] + ): 788.1904; found 778.1913.

[0122] Example 26: Synthesis of compound 9a was synthesized from compound 4, the specific synthesis steps were referred to Example 2.

[0123] Compound 9a was a white powder; yield: 44 %; melting point: 85.4-88.7 °C;

[0124] 1 H NMR (400 MHz, CDCl3): δ (ppm)8.54 (s, 1H, Amino-H), 7.25 (d, J = 9.6Hz, 1H, H30), 7.16 (t, J= 2.8 Hz, 1H, H35), 7.11 (d, J = 2.4 Hz, 1H, H36), 6.84(dd, J = 8.8, 2.4 Hz, 1H, H33), 6.54 – 6.37 (m, 2H, H19, H32), 5.77 (d, J = 8.4Hz, 1H, H14), 5.31 (dd, J = 11.2, 1.2 Hz, 1H, H20), 5.18 (dd, J = 17.2, 1.2 Hz,1H, H20), 4.68 (s, 2H, H28), 3.74 – 3.58 (m, 4H, Piperazinyl-H), 3.39 – 3.28(m, 1H, H11), 3.21 – 3.11 (m, 1H, H22), 3.10 – 2.99 (m, 1H, H22), 2.66 – 2.42(m, 4H, Piperazinyl-H), 2.39 – 1.98 (m, 6H, H2,H4, H10, H13), 1.82 – 1.30(m, 9H, H1, H6, H7, H8, H15), 1.15 (s, 3H, H18), 1.12 – 1.05 (m, 1H, H8),0.87 (d, J = 6.8 Hz, 3H, H17), 0.69 (d, J = 6.8 Hz, 3H, H16). 13C NMR (101 MHz, CDCl3): δ (ppm) 217.1 (C3), 168.9 (C21), 167.1 (C4), 152.3 (C29), 139.1 (C19), 131.6 (C34), 128.3 (C31), 125.3 (C33), 117.2 (C20), 112.3 (C36), 111.9 (C35), 104.0 (C30), 102.3 (C32), 74.6 (C11), 68.6 (C14), 59.7 (Piperazinyl-C), 58.2 (Piperazinyl-C), 53.0 (Piperazinyl-C), 52.5 (Piperazinyl-C), 45.5 (C22), 45.1 (C9), 44.0 (C13), 41.9 (C5), 36.7 (C6), 36.1 (C19), 34.5 (C2), 30.4 (C8), 26.8 (C7), 26.5 (C18), 24.9 (C1), 16.7 (C16), 14.9 (C15), 11.5 (C17). IR (KBr, cm -1 ) : 3424, 2919, 1731, 1639, 1459, 1401, 1384, 1289, 1216, 1157, 1117, 1010. HRMS: calculated for C 36 H 49 N3O6([M + H] + ): 620.3694; found 620.3700.

[0125] Test detection

[0126] One: minimum inhibitory concentration (MIC) activity study

[0127] MIC test method

[0128] 1. Experimental strain: Staphylococcus aureus drug-resistant strain ATCC 33591 and ATCC 43300, Staphylococcus aureus sensitive strain ATCC 29213, Staphylococcus epidermidis drug-resistant strain ATCC 51625, Staphylococcus epidermidis sensitive strain ATCC 12228 and Escherichia coli standard strain ATCC 25922 were selected as MIC value determination strains.

[0129] 2. Drug dilution: the target compound and tylosin (T) were dissolved and diluted with DMSO as solvent, and prepared into a concentration of 12800μg •mL -1 The mother liquor was placed in a refrigerator, sealed in the dark, and stored for standby use.

[0130] 3. Preparation of bacterial solution: The test bacteria were activated, and single colonies were selected and placed in 0.9% saline to prepare a bacterial solution at a McFarland concentration of 0.5 (1.5 x 10 8 CFU•mL -1 The solution was then diluted 10 times with Mueller-Hinton sterile broth medium (MHB) for standby use.

[0131] 4. Positive control: Tylosin was selected as the positive control.

[0132] 5. MIC / MBC determination: In a 96-well plate, 100 μ L MHB was added to the remaining wells except the edge wells and the second column of wells, 198 μ L MHB and 2 μ L mother liquor were added to the second column of wells. The compound and the positive control were diluted using a two-fold dilution method, and a total of 64 – 0.03125 μg •mL -1 12 different concentration gradients of the dilutions were prepared, 100 μ L bacterial solution was added to each well except the edge wells, and the mixture was thoroughly mixed. The mixture was incubated at 37°C for 18 – 24 h, and the growth of the test bacteria was observed. The lowest concentration of the drug without growth was taken as the MIC value of the drug for the test bacteria. Tylosin was used as the positive control, and an ethanol solution with the same concentration as the prepared compound was used as the negative control. Each test bacteria was subjected to three parallel experiments, and the experiments were repeated three times. The growth of the test bacteria in the negative control group was good, and the results of the remaining experiments are shown in Table 1.

[0133] Table 1 MIC of piperazine-linked benzoheterocyclic leptomycin derivatives (5a-9a)

[0134] Cpd (g / mL) MRSA ATCC 33591 MRSA ATCC 4 3300 MSSA ATCC 29213 MRSE ATCC 51625 MSSE ATCC 12228 E. coli ATCC 25922 5a　 0.25 0.25 0.25 0.0625 0.0625 >64 5b 0.25 0.25 0.25 0.0625 0.015625 >64 5c 0.125 0.25 0.25 0.03125 0.0625 >64 5d 0.25 0.5 0.5 0.0625 0.125 >64 5e 0.25 0.25 0.25 0.0625 0.125 >64 5f 0.25 0.25 0.25 0.03125 0.0625 >64 5g 0.25 0.25 0.25 0.0625 0.125 >64 5h 0.25 0.5 0.5 0.125 0.0625 >64 5i 0.25 0.25 0.25 0.03125 0.0625 >64 5j 0.25 0.5 0.5 0.03125 0.0625 >64 5k 0.25 0.25 0.25 0.03125 0.03125 >64 5l 0.25 0.5 0.25 0.125 0.0625 >64 6a 0.0625 0.125 0.125 0.03125 0.03125 >64 7a 0.0625 0.125 0.125 0.0625 0.03125 >64 8a 0.25 0.25 0.125 0.0625 0.125 >64 8b 0.25 0.25 0.25 0.03125 0.125 >64 8c 0.25 1 2 0.015625 0.0625 >64 8d 0.125 0.25 0.125 0.015625 0.03125 >64 8e 0.25 0.25 0.25 0.125 0.03125 >64 8f 0.125 0.25 0.25 0.015625 0.015625 >64 8g 0.25 0.5 0.5 0.125 0.0625 >64 8h 0.25 0.25 0.25 0.125 0.015625 >64 8i 0.25 0.25 0.25 0.125 0.0625 >64 9a 0.25 0.25 0.25 0.03125 0.03125 >64 T 0.5 0.5 1 0.125 0.125 32

[0135] As can be seen from Table 1, the compounds of the present application exhibit excellent antibacterial effects on the gram-positive bacteria Staphylococcus aureus resistant strains ATCC33591 and ATCC43300, Staphylococcus aureus sensitive strain ATCC 29213, Staphylococcus epidermidis resistant strain ATCC 51625, and Staphylococcus epidermidis sensitive strain ATCC 12228, especially compounds 5a-7a, which have a MIC of 0.3125-0.5 μg •mL -1, all of which are better than tiamulin. However, the compounds do not show good antibacterial activity against gram-negative bacteria Escherichia coli standard strain ATCC 25922. In summary, the piperazine-pleuromutilin compounds in the present application show excellent antibacterial effect against gram-positive bacteria, and are expected to treat bacterial infections caused by gram-positive bacteria.

[0136] II: Bacteriostatic curve activity research

[0137] Experimental method

[0138] In this experiment, the test bacteria were recovered and subcultured, and the bacterial liquid concentration was adjusted to about 0.6 McFarland by normal saline, and then diluted 10 3 times with Mueller-Hinton sterile broth medium (MHB). Compound 6a and tiamulin were prepared into working solutions with concentrations of 2xMIC, 4xMIC and 8xMIC, and an equal volume of prepared bacterial liquid was added to make the final concentration of the test compound 1xMIC, 2xMIC and 4xMIC, respectively. A bacterial growth curve without the test compound was prepared as a control. At 0h, 2h, 4h, 6h, 8h, 12h and 24h, 100 μ L of each concentration was sampled, diluted 100 times with normal saline, and 100 μ L of the diluted sample was evenly coated on MHA plates, which were further incubated at 37°C for 20h, and three groups of parallel controls were set. The total number of bacteria on the plates (CFU·mL -1 ) was determined. 10 CFU·mL -1 vs. time was plotted to draw the bactericidal curve.

[0139] The experimental data are shown in the following figure ( Figure 1 ). The bacterial growth trend of compound 6a and tiamulin is similar, and the bacteria in the blank control group are in exponential growth during the growth process. At a concentration of 1xMIC and 2xMIC, the bacterial growth can be inhibited compared with the blank control group, and the growth rate is significantly decreased. The inhibitory effect of compound 6a is slightly better than that of tiamulin. At a concentration of 4xMIC, in the presence of tiamulin, a decreasing trend can be observed at 0-12h, and the decreasing trend is obvious at 12-24h. At 24h, the bacteria can be basically inhibited. However, the bacterial growth can be basically inhibited at 8-12h in the presence of compound 6a, which is consistent with the MBC value of compound 6a measured by us.

[0140] III: Mouse thigh tissue infection model experiment

[0141] Experimental method

[0142] Determination of the optimal incubation conditions of testosterone: change the concentration of testosterone (1000, 2000, 3000, 4000, 5000, 6000, 7000 μ mol / L), try to find the optimal incubation conditions of testosterone by measuring the metabolic amount of testosterone, when the metabolic amount of testosterone is near 20%, it is the optimal incubation conditions of testosterone. Add 4 μ L SD rat liver microsomes (final concentration 0.2 mg / mL) to 182 μ L PBS buffer solution, respectively add 2 μ L testosterone solution of different concentrations (final concentration 10, 20, 30, 40, 50, 60, 70 μ mol / L) (the operation process is carried out on the ice bath at 4 o C), mix well, then pre-incubate at 37 o C water bath for 10 min, then add NADPH regeneration system (NADPH also needs to be pre-incubated for 5 min), start the reaction, incubate at 37 o C water bath for 20 min, then add 200 μ L pre-cooled methanol to terminate the reaction. The sample is centrifuged at 4 o C at 15000 r / min for 20 min, and the supernatant is taken to measure HPLC, the sample is 10 μ L. The metabolic amount of testosterone in the experimental group is measured by HPLC. Set the concentration of testosterone as C0, the peak area of testosterone metabolite of different concentrations corresponds to the concentration of testosterone C x in the standard curve, and the metabolic amount of testosterone is calculated as follows: X = (C0-C x ) / C0x100%. Since the optimal incubation conditions cannot be found by changing the concentration of testosterone only, the most suitable concentration of testosterone in the previous step is determined by changing the volume of liver microsomes, pre-incubation time and incubation time to determine the optimal incubation conditions of testosterone (set a single variable).

[0143] The preparation of the incubation system is as follows: add 2 μ L SD rat liver microsomes (final concentration 0.1 mg / mL) to 182 μ L PBS buffer solution, add 2 μ L testosterone solution and 2 μ L compound 6a solution (concentration is 0, 0.2, 0.5, 1.5, 10, 20, 50 μ mol / L), pre-incubate at 37 o C for 5 min, then add NADPH regeneration system 12 μ L to start the reaction, incubate at 37 o C for 20 min, then add 200 μ L pre-cooled methanol to terminate the reaction, 4 oC, 15000 r / min centrifugal 20 min, take the supernatant to measure HPLC. The metabolic amount of probe substrate in each test group was determined by HPLC, and the metabolic amount of the control group was V0, and the metabolic amount of the rest of the different concentrations was Vx, and then the residual enzyme activity I = (1-Vx / V0) x 100% was calculated.

[0144] The experimental results are shown in Figure ( Figure 2 ) as shown in the figure, it can be seen that the inhibitory effect of compound 6a on CYP3A4 enzyme in rat liver microsomes increases with the increase of the concentration of compound 6a, and the IC50 value is 24.32 μ M. The results show that compound 6a has weak inhibitory activity on CYP3A4 enzyme, and the MBC of compound 6a is only 0.25 μg / mL, at a lower use concentration, it will not affect the drug relying on CYP3A4 enzyme metabolism.

[0145] Four: Experimental study on thigh tissue infection model of mice

[0146] Experimental method

[0147] Randomly select 9 SPF level 5-6 week old female ICR mice, on the first day of the experiment, intraperitoneally inject cyclophosphamide at a dose of 150 mg / kg, on the fourth day, intraperitoneally inject cyclophosphamide at a dose of 100 mg / kg, and collect blood from the posterior orbital venous plexus of the mouse. Determine whether the number of neutrophils in the blood of the mouse is less than 100 / mm 3 , if the number of neutrophils is less than 100 / mm 3 , the mouse reaches immunosuppression, then the follow-up experiment can be carried out. A single MASA ATCC 33591 colony is inoculated in a 4 mL MHB broth test tube, and placed in a 37 o C constant temperature incubator for 12 h, then placed in a 37 o C, 210 rpm constant temperature incubator for 0.5 h, so that the bacteria grow to the logarithmic phase, and the bacterial solution is diluted with sterile saline, and the bacterial amount is adjusted to 10 7 CFU / mL for standby. The immunosuppressed mice are randomly divided into 3 groups according to only inoculating bacterial solution, tylosin treatment group and compound 6a treatment group, 3 mice in each group, 0.1 mL of prepared 10 7CFU / mL of bacteria solution was injected into the thigh muscle of neutropenic mice, 2 h later, tail vein injection was given for treatment, the blank control group was injected with 10 mL / kg solvent of the test compound. 24 h after administration, the mice were sacrificed by CO2 asphyxiation, and the thigh muscles of the mice were immediately separated and placed in pre-cooled 3 mL sterile normal saline, and homogenized with a sterilized tissue homogenizer. After homogenization, 0.1 mL of the slurry was diluted in 10 times the amount of sterile normal saline, and 25 μ L dilution was plated on MHA agar plates for counting, and the bacterial load per gram of muscle was calculated based on the colony count results.

[0148] The results of the experiment are shown in Figure ( Figure 3 ) Compared with the no-drug control group, both Retapamulin (40 mg / kg) and compound 6a (40 mg / kg) can reduce the bacterial load in the thigh, and the same dose of compound 6a can have a matching antibacterial effect as Retapamulin, and it can be seen that the bacterial load in the thigh of mice after treatment with compound 6a is slightly lower than that after treatment with Retapamulin. The results show that compound 6a has effective in vivo anti-MRSA activity.

[0149] Five, experimental study of mouse MRSA systemic infection model

[0150] Experimental method

[0151] The solutions were prepared as follows: for the tyrothricin solution, 10 mg of tyrothricin was weighed and dissolved in 10 mL of water to prepare a 1 mg / mL stock solution, which was stored at room temperature; for the compound 6a solution, 31 mg of compound was dissolved in 12.4 mL of 10% dimethyl sulfoxide to prepare a 2.5 mg / mL solution; finally, 262.5 mg of cyclophosphamide was dissolved in 7 mL of water to prepare a 37.5 mg / mL solution, which was stored at -20°C. Then the concentration was adjusted according to the needs of intraperitoneal injection. Kunming female mice with a body weight of about 22-25 g were randomly selected, 9 mice per group, 150 and 100 mg / kg of cyclophosphamide were injected into the abdominal cavity of the mice on the 1st and 4th days of the experiment, 24 h after the last cyclophosphamide injection, the mice were anesthetized with isoflurane, and 0.4 mL of 10 8 CFU / mL MRSA 33591 MRSA 33591 infection model was established. 1 h after infection, 0.1 mL / 10 g of tail vein injection was given for administration, and different concentrations of compound 6a (5, 10, 20, 40 mg / kg) and tyrothricin (5, 10, 20, 40 mg / kg) were injected in the experimental groups, and the same volume of 0.9% normal saline was injected in the control group. Observe twice a day, record the symptoms and death, and take the survival rate of mice 7 days after infection as the endpoint.

[0152] To determine the tissue distribution of MRSA and perform histological analysis, surviving mice were anesthetized with excess CO2 and euthanized at the endpoint. The liver, kidney, spleen, lung and heart of euthanized mice and the liver, kidney, spleen, lung and heart of mice that died during the treatment were collected, weighed and ground into sterile saline homogenate. Then it was diluted 10 times. The resulting dilution was spread on MH agar plates and incubated at 37 °C for 24 h. This procedure was performed in triplicate. The viable colony counts were enumerated and expressed as colony forming units (CFU / mL). In addition, tissue specimens were fixed with 10% formalin, paraffin-embedded, sectioned at 4 μ m thickness, HE stained and observed by confocal microscopy.

[0153] The experimental results are shown in FIGS. Figure 4 and Figure 5 , in the MRSA systemic infection mouse model, the positive control group of mice infected with MRSA showed clinical symptoms, and the survival rate at the endpoint was only 10%; while after 6a or teixobactin treatment for 7 days, both showed dose-dependent protective effects. The survival rates of 6a treatment group mice at doses of 20 mg / kg and 40 mg / kg were 50% and 70% respectively, which were significantly higher than those of the same dose of valnemulin treatment group (40% and 60% respectively). The results of this in vivo efficacy experiment show that compound 6a is a new candidate drug for treating MRSA infection.

[0154] The analysis of bacterial load in the main organs (heart, liver, spleen, lung, kidney) of systemic MRSA ATCC 33591 infected mice after treatment with compound 6a and teixobactin (40 mg / kg) is shown in Figure 6 As compared with the control group, the bacterial load in the heart, liver, spleen, lung and kidney of mice in the compound 6a treatment group was significantly reduced by 2, 1.29, 1.53, 2.2 and 2.1 log 10 CFU / mL respectively; while the reduction in the corresponding organs of the teixobactin treatment group was 2, 0.95, 1.34, 1.49 and 1.57 log 10 CFU / mL respectively. It is worth noting that compound 6a showed better antibacterial effect than teixobactin in all the tested organs, especially in the lung and kidney, showing more significant antibacterial activity (the reduction was 0.71 and 0.53 log 10 CFU / mL higher respectively). These data fully demonstrate that compound 6a has excellent therapeutic effect on systemic MRSA infection in mice, and its antibacterial efficacy significantly surpasses that of the clinically commonly used drug teixobactin.

[0155] The results of H&E staining of organ tissue sections of MRSA ATCC 33591 infected mice are shown inFigure 7 As shown, compared with the normal control group, the mice in the MRSA infection group showed obvious pathological damage in each organ—myocardial fibers were sparse, local myocardial cells were hypertrophic with reduced number, interstitial connective tissue was slightly proliferated and inflammatory infiltrated; liver parenchyma was diffusely damaged, mononuclear cell infiltrates were visible in the intercellular space of liver cells and around blood vessels, and local aggregated inflammatory plaques were formed; the number of white pulp in spleen tissue was reduced, the boundary of marginal zone was blurred, and a small amount of atrophic cells were distributed; lung tissue damage was more obvious, pulmonary interstitium was widely thickened, and the pulmonary alveoli were disappeared; a large number of inflammatory cells were aggregated in kidney tissue, and part of the cell nuclei were fragmented. Notably, the organ pathological damage of the mice in the compound 6a treatment group was significantly reduced, showing reduced inflammatory cell infiltration, tissue structure remodeling and cell morphology recovery, and the treatment effect was better than that of the positive control drug thymosin. The above results show that compound 6a has a repairing effect on the multiple organ damage caused by MRSA ATCC 33591 infection, and has potential value as a candidate drug for anti-MRSA infection.

[0156] The above specific embodiments further illustrate the purpose, technical solutions and beneficial effects of the present application. It should be understood that the above description is only a specific embodiment of the present application and is not intended to limit the protection scope of the present application. Any modification, equivalent replacement, improvement, etc. within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. A heterocyclic piperazine truncated pleurotin derivative, characterized in that, This includes compounds with structures as shown in Formula I, or their stereoisomers or pharmaceutically acceptable salts. Formula I Where R is selected from , , and One of them; R1 is selected from one of H, F, Cl, Br, CH3, OCH3, NO2 and NHBoc, and R2 is selected from one of H, F, Cl, Br, CH3 or OCH3; R3 is H; R4 is H.

2. The heterocyclic piperazine truncated pleurotin derivative according to claim 1, characterized in that, The pharmaceutically acceptable salts include one of the following: hydrochloride, fumarate, malate, hydrobromide, succinate, phosphate, methanesulfonate, and benzoate.

3. A heterocyclic piperazine truncated pleurotin derivative, characterized in that, The derivatives include compounds with one of the following structures: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 。 4. The use of a heterocyclic piperazine truncated pleurotin derivative as described in any one of claims 1-3 in the preparation of a medicament for treating infectious diseases.

5. The application according to claim 4, characterized in that, The infectious diseases mentioned include those caused by mycoplasma or drug-resistant bacteria.

6. The application according to claim 5, characterized in that, This includes administering the heterocyclic piperazine truncated pleurotin derivative in combination with pharmaceutically acceptable excipients.

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