Targeting sirt3 agonists and their use in anti-tumor therapeutics

Abstract

The present application relates to a SIRT3 targeting agonist and its application in anti-tumor therapeutic drugs, and belongs to the technical field of tumor therapeutic drug discovery. The compound of the present application or a pharmaceutically acceptable salt or isomer structure thereof is shown as formula I, wherein Ar and R are as described in the claims and the specification. The compound described in the present application or a pharmaceutically acceptable salt or isomer thereof and a pharmaceutical composition thereof have SIRT3 agonistic activity. And have obvious anti-tumor effect, especially for breast cancer, the inhibitory effect is remarkable, and can be used for preparing an anti-tumor drug.

Description

Technical Field

[0001] This invention relates to SIRT3-targeting agonists and their application in antitumor therapeutics, belonging to the field of tumor therapeutic drug discovery technology. Background Technology

[0002] Cancer is one of the leading causes of death worldwide in recent years, and its threat to humanity is becoming increasingly prominent. Globally, there are approximately 20 million new cancer cases and 9.7 million cancer deaths, meaning about one-fifth of people will develop cancer in their lifetime, making it a major public health issue worldwide. Current cancer treatments include surgical resection, radiotherapy, chemotherapy, targeted drug therapy, and immunotherapy. Drug therapy has become an effective and widely used treatment for malignant tumors. Clinically used anti-tumor drugs are numerous, including those that directly act on DNA, interfere with DNA synthesis, inhibit cell mitosis, interfere with tumor signal transduction, and are novel anti-tumor drugs. Examples include paclitaxel, anthracyclines, and platinum compounds.

[0003] In the field of malignant tumor drug therapy, discovering new therapeutic targets, designing novel targeted small molecule drugs, and deeply exploring their mechanisms of action are key to solving clinical problems and have significant research significance and value. SIRT3, as an important oncogene, is abnormally underexpressed in malignant tumors. Designing targeted small molecule agonists to mediate metabolic reprogramming of tumor cells is an important strategy for treating breast cancer, especially triple-negative breast cancer. Summary of the Invention

[0004] The technical problem solved by this invention is to provide a compound that can be used as a SIRT3 agonist.

[0005] This invention provides compounds with the structural formula shown in Formula I, or pharmaceutically acceptable salts or isomers thereof:

[0006]

[0007] Wherein, Ar is a 6-10 aryl or heteroaryl, and the heteroaryl contains 1-3 heteroatoms of N, O or S;

[0008] The Ar is substituted by one or more of the following substituents: H, cyano, C1-C6 alkoxy, C1-C6 alkyl, halogen, C1-C6 ester, halo-C1-C6 alkyl, phenyl, benzyloxy, methyl sulfone;

[0009] R represents substituted or unsubstituted phenyl, naphthyl, thiophene, or furanyl groups;

[0010] The substituents are C1-C6 alkoxy, C1-C6 alkyl, halogen, or halogenated C1-C6 alkyl;

[0011] The present invention preferably contains compounds with the following structures or pharmaceutically acceptable salts or isomers thereof:

[0012] Wherein, Ar is naphthyl, 2,3-methylenedioxy-phenyl, pyridyl, or phenyl;

[0013] The Ar is substituted by one or more of the following substituents: H, cyano, C1-C6 alkoxy, C1-C6 alkyl, halogen, halo-C1-C4 alkyl, C1-C6 ester, phenyl, benzyloxy, methyl sulfone;

[0014] R represents substituted or unsubstituted phenyl, naphthyl, thiophene, or furanyl groups;

[0015] The substituents are C1-C4 alkoxy, C1-C4 alkyl, halo-C1-C4 alkyl, or halogen.

[0016] The present invention preferably contains compounds with the following structures or pharmaceutically acceptable salts or isomers thereof:

[0017] Wherein, Ar is naphthyl, 2,3-methylenedioxy-phenyl, 4,5-dichloropyridyl,

[0018] X1 is hydrogen, phenyl, C1-C6 alkyl, C1-C4 alkoxy, F, Cl, Br, I, benzyloxy, CF3, methylsulfonyl, cyano, or methoxyformyl.

[0019] R is Naphthyl, thiophenyl, furanyl;

[0020] X2 can be hydrogen, C1-C4 alkoxy, C1-C4 alkyl, halogenated C1-C4 alkyl, or halogen.

[0021] The present invention preferably contains compounds with the following structures or pharmaceutically acceptable salts or isomers thereof:

[0022] Wherein, Ar is naphthyl, 2,3-methylenedioxy-phenyl, 4,5-dichloropyridyl, or...

[0023] X1 represents single or multiple substitutions at positions 2-5;

[0024] R is

[0025] X2 represents 3-5 positions of single or multiple substitutions.

[0026] The present invention preferably contains compounds with the following structures or pharmaceutically acceptable salts or isomers thereof:

[0027] Wherein, Ar is 4,5-dichloropyridinyl;

[0028] R is naphthyl, thiophene, furanyl, or other compounds.

[0029] X2 is a C1-C4 alkoxy, C1-C4 alkyl, halo-C1-C4 alkyl, or halogen.

[0030] The present invention preferably contains compounds with the following structures or pharmaceutically acceptable salts or isomers thereof:

[0031] Where Ar is X1 is a methyl sulfone group;

[0032] R is Naphthyl, thiophene, furanyl, X2 is hydrogen, C1-C4 alkoxy, C1-C4 alkyl, CF3, Br, Cl.

[0033] The present invention preferably contains compounds with the following structures or pharmaceutically acceptable salts or isomers thereof:

[0034]

[0035]

[0036]

[0037] The pharmaceutically acceptable salts of the compounds described in this invention can be nitrates, hydrochlorides, sulfates, phosphates, or citrates of the compounds.

[0038] The present invention also provides the use of the above-mentioned compounds or their pharmaceutically acceptable salts or isomers in the preparation of SIRT3 agonists.

[0039] The present invention also provides the use of the above-mentioned compounds or their pharmaceutically acceptable salts or isomers in the preparation of antitumor drugs.

[0040] Furthermore, the compounds described in this invention, or their pharmaceutically acceptable salts or isomers, can be used as SIRT3 agonists for the treatment of tumors and related diseases.

[0041] The tumor is breast cancer, and the breast cancer is further described as triple-negative breast cancer.

[0042] This invention also provides a pharmaceutical composition comprising an effective dose of the above-described compound or its pharmaceutically acceptable salts or isomers. The compounds of this invention can be formulated into the following forms by methods known in the art: tablets, capsules, aqueous or oily solutions, suspensions, emulsions, creams, ointments, gels, nasal sprays, suppositories, finely dispersed powders or aerosols or sprays for inhalation, and sterile aqueous or oily solutions or suspensions or sterile emulsions for parenteral (including intravenous, intramuscular, or infusion) administration. Liquid formulations can be prepared using sterile water or a water-propylene glycol solution as a solvent, and the active ingredient can also be formulated in an aqueous polyethylene glycol solution. Aqueous solutions for oral administration can be prepared by dissolving the active ingredient in water and adding suitable colorants, flavoring agents, stabilizers, and thickeners as needed. Orally administered aqueous suspensions can be made by dispersing finely dispersed active components in water along with a viscous substance, such as natural synthetic gums, resins, methylcellulose, carboxymethylcellulose, and other suspending agents known in the pharmaceutical field.

[0043] The pharmaceutical composition may be in unit dose form. In these forms, the composition is divided into unit doses containing an appropriate amount of the active ingredient. The unit dose form may be a packaged formulation comprising a portion of the formulation, such as boxed tablets, capsules, and powders in tubular vials or ampoules. The unit dose form may also be capsules or tablets, or any of these packaging forms in appropriate quantities.

[0044] The active ingredient of the pharmaceutical composition of the present invention may be the compound of the present invention alone, or it may be combined with other antitumor compounds as the active ingredient.

[0045] In the treatment of tumors, the compounds of the present invention or their pharmaceutically acceptable salts, isomers or pharmaceutical compositions may be used in combination with other antitumor drugs.

[0046] In the treatment of tumors, such combination therapy can be achieved by administering various therapeutic components simultaneously, sequentially, or individually. Such combination products utilize compounds of the present invention within their effective dose range and other pharmaceutically active agents within their permitted dose range. Attached Figure Description

[0047] Figure 1 The proliferation of BT-549 and MDA-MB-231 cells after treatment with compound 42 for 6, 12, 24, 36 and 48 hours was detected by MTT assay.

[0048] Figure 2 To detect the proliferation of BT-549 and MDA-MB-231 cells treated with compound 42 for 24 and 48 hours in a 3D tumor spheroid assay.

[0049] Figure 3To detect DNA replication in BT-549 and MDA-MB-231 cells treated with 0 μM and 2 μM compound 42 using the EdU assay.

[0050] Figure 4 The cell migration of BT-549 cells treated with 0 μM, 1 μM, 2 μM and 4 μM compound 42 for 24 hours and 48 hours was detected by scratch assay.

[0051] Figure 5 The cell migration of MDA-MB-231 cells treated with 0 μM, 1 μM, 2 μM and 4 μM compound 42 for 24 hours and 48 hours was detected by scratch assay.

[0052] Figure 6 To detect glucose uptake and lactose production in BT-549 and MDA-MB-231 cells treated with 0 μM, 1 μM, 2 μM and 4 μM compound 42 by flow cytometry.

[0053] Figure 7 To detect mitochondrial membrane potential and ATP production in BT-549 and MDA-MB-231 cells treated with 0 μM, 1 μM, 2 μM and 4 μM compound 42 by flow cytometry.

[0054] Figure 8 To detect apoptosis in BT-549 and MDA-MB-231 cells treated with 0 μM, 1 μM, 2 μM and 4 μM compound 42 by flow cytometry.

[0055] Figure 9 To investigate the in vivo antitumor activity of compound 42 in a nude mouse model with a BT-549 xenograft.

[0056] Figure 10 Daily body weight and organ ratio of animals in the BT-549 xenograft nude mouse model. Detailed Implementation

[0057] The specific embodiments of the present invention will be further described below with reference to examples, but the present invention is not limited to the scope of the embodiments described herein.

[0058] Example 1: Synthesis of compounds 1-50

[0059] Compounds 1–50 were synthesized using the following reaction formula:

[0060]

[0061] Synthetic route and conditions: (I) Glacial acetic acid, refluxed at 80°C for 8 hours; (II) DIEA, dissolved in dichloromethane, reacted at room temperature for 10 hours; where Ar and R are as described above.

[0062] Preparation of intermediate 1-23a

[0063] Different substituted aromatic aldehydes or heteroaromatic aldehydes (8.92 mmol), ethyl propynate (17.84 mmol), and ammonium acetate (8.92 mmol) were dissolved in glacial acetic acid (5 mL), and the mixture was stirred under reflux at 80 °C for 8 h. The reaction progress was monitored by TLC. The reaction was stopped, the reaction flask was brought to room temperature, 50 mL of water was added to quench the reaction, and an appropriate amount of sodium hydroxide solid was slowly added while stirring to adjust the pH of the reaction solution to 7-8. The mixture was then stirred at room temperature for 10 min. The solution was extracted with ethyl acetate (3 × 30 mL), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a brown crude product. This crude product was purified by silica gel column chromatography in a PE:EA = 3:1 elution system to give intermediate 1-23a, a yellow solid, in a yield of 30-40%.

[0064] Intermediate 1a, 1 H-NMR (400MHz, CDCl3), δ (ppm): 7.55 (d, 2H, J = 8.33Hz), 7.46 (d, 2H, J = 8.33Hz), 7.37 ( d,2H,J=5.31Hz),6.53(s,1H),4.97(s,1H),4.03-4.13(m,4H),1.18(t,6H,J=7.14Hz), 13 C-NMR (100MHz, CDCl3), δ (ppm): 166.6 (2C), 152.0, 134.0 (2C), 131.9 (2C), 129.2 (2C), 119.2, 110.1, 107.7 (2C), 60.3 (2C), 38.2, 14.2 (2C).

[0065] Intermediate 2a, 1 H-NMR (400MHz, CDCl3), δ (ppm): 7.28 (d, 2H, J = 5.35Hz), 7.25 (d, 2H, J = 8.65Hz), 6.86 (brs, 1H) ,6.78(d,2H,J=8.65Hz),4.83(s,1H),4.02-4.13(m,4H),3.74(s,3H),1.20(t,6H,J=7.20Hz), 13 C-NMR (100MHz, CDCl3), δ (ppm): 167.4 (2C), 158.1, 139.6, 133.7 (2C), 129.3 (2C), 113.3 (2C), 108.4 (2C), 60.0 (2C), 55.1, 36.7, 14.2 (2C).

[0066] Intermediate 3a,1 H-NMR (400MHz, CDCl3), δ (ppm): 7.32 (d, 2H, J = 5.30Hz), 7.23 (d, 2H, J = 8.00Hz), 7.05 (d, 2H, J = 8.00Hz),6.62(brs,1H),4.86(s,1H),4.03-4.13(m,4H),2.28(s,3H),1.19(t,6H,J=7.12Hz), 13 C-NMR (100MHz, CDCl3), δ (ppm): 167.0 (2C), 144.0, 133.3 (2C), 128.7 (3C), 128.2 (2C), 108.9 (2C), 60.0 (2C), 39.1, 21.1, 14.2 (2C).

[0067] Intermediate 4a, 1 H-NMR (400MHz, CDCl3), δ (ppm): 7.36 (d, 2H, J = 8.40Hz), 7.32 (d, 2H, J = 5.32Hz), 7.22 ( d,2H,J=8.40Hz),6.75(s,1H),4.87(s,1H),4.03-4.13(m,4H),1.19(t,6H,J=7.14Hz), 13 C-NMR (100MHz, CDCl3), δ (ppm): 167.0 (2C), 146.0, 133.9 (2C), 131.0 (2C), 130.1 (2C), 120.3, 108.1 (2C), 60.2 (2C), 37.3, 14.2 (2C).

[0068] Intermediate 5a, 1 H-NMR (400MHz, CDCl3), δ (ppm): 7.31 (d, 2H, J = 5.26Hz), 6.78-6.83 (m, 1H), 6.72 (t, 1H, J = 7.69Hz), 6. 62-6.83(m,1H),6.22(brs,1H),5.95(s,2H),5.02(s,1H),4.02-4.14(m,4H),1.19(t,6H,J=7.15Hz), 13 C-NMR (100MHz, CDCl3), δ (ppm): 167.0 (2C), 147.1, 145.1, 133.8 (2C), 128.6, 123.0, 121.0, 107.1, 106.6, 100.6, 60.0 (2C), 33.6, 14.2 (2C).

[0069] Intermediate 6a, 1H-NMR (400MHz, CDCl3), δ (ppm): 7.92 (d, 2H, J = 8.36Hz), 7.42 (d, 2H, J = 8.36Hz), 7.36 (d, 2H, J = 5.32Hz),6.44(brs,1H),4.97(s,1H),4.00-4.12(m,4H),3.88(s,3H),1.18(t,6H,J=7.19Hz), 13 C-NMR (100MHz, CDCl3), δ (ppm): 167.2, 166.8 (2C), 151.9, 133.8 (2C), 129.4 (2C), 128.5 (2C), 128.2, 108.2 (2C), 60.2 (2C), 52.0, 37.9, 14.2 (2C).

[0070] Intermediate 7a, 1 H-NMR (400MHz, CDCl3), δ (ppm): 7.56 (d, 2H, J = 8.27Hz), 7.32 (d, 2H, J = 5.26Hz), 7. 09(d,2H,J=8.27Hz),6.47(brs,1H),4.85(s,1H),4.03-4.13(m,4H),1.19(t,6H,J=

[0071] 7.17Hz), 13 C-NMR (100MHz, CDCl3), δ (ppm): 166.9 (2C), 146.6, 137.0 (2C), 133.7 (2C), 130.5 (2C), 108.2 (2C), 92.0, 60.2 (2C), 37.4, 14.2 (2C).

[0072] Intermediate 8a, 1 H-NMR (400MHz, CDCl3), δ (ppm): 8.26 (d, 1H, J = 2.16Hz), 7.72 (d, 1H, J = 2.16Hz), 7. 38(d,2H,J=5.33Hz),6.59(brs,1H),4.94(s,1H),4.07-4.15(m,4H),1.22(t,6H,J=

[0073] 7.08Hz), 13 C-NMR (100MHz, CDCl3), δ (ppm): 166.2 (2C), 147.2 (2C), 142.7, 138.8, 134.3 (3C), 107.1 (2C), 60.5 (2C), 35.0, 14.2 (2C).

[0074] Intermediate 9a,1 H-NMR (400 MHz, CDCl₃), δ (ppm): 7.70-7.80 (m, 4H), 7.50-7.57 (m, 1H), 7.71-7.75 (m, 2H), 7.37-7.43 (m, 2H), 7.36 (d, 2H, J=5.33 Hz), 6.42 (brs, 1H), 5.07 (s, 1H), 4.00-4.09 (m, 4H), 1.17 (t, 6H, J=7.10 Hz), 13 C-NMR (100 MHz, CDCl₃), δ (ppm): 167.0 (2C), 144.5, 133.4 (2C), 132.4, 127.9, 127.5 (2C), 127.2, 126.9, 126.4, 125.6, 125.3, 108.8 (2C), 60.1 (2C), 37.8, 14.2 (2C). Intermediate 10a, 1 H-NMR (400 MHz, CDCl₃), δ (ppm): 7.31 (d, 2H, J=5.26 Hz), 6.98 (dd, 1H, J=3.17 Hz), 6.78-6.83 (m, 1H), 6.74 (dd, 1H, J=4.54 Hz), 6.28 (brs, 1H), 5.20 (s, 1H), 4.00-4.11 (m, 4H), 3.83 (s, 3H), 1.17 (t, 6H, J=7.09 Hz), 13 C-NMR (100 MHz, CDCl₃), δ (ppm): 167.0 (2C), 153.3, 134.1 (2C), 117.6, 117.4, 113.7, 113.4, 111.7, 108.0 (2C), 59.9 (2C), 56.3, 32.5, 14.2 (2C).

[0075] Intermediate 11a, 1 H-NMR (400 MHz, CDCl₃), δ (ppm): 7.31-7.34 (m, 4H), 7.23 (d, 2H, J=7.47 Hz), 7.14 (t, 1H, J=6.92 Hz), 6.58 (brs, 1H), 4.89 (s, 1H), 4.02-4.13 (m, 4H), 1.19 (t, 6H, J=7.16 Hz), 13 C-NMR (100 MHz, CDCl₃), δ (ppm): 167.2 (2C), 146.9, 133.7 (2C), 128.3 (2C), 128.0 (2C), 126.4, 108.5 (2C), 60.1 (2C), 37.6, 14.2 (2C).

[0076] Intermediate 12a, 1 H-NMR (400MHz, CDCl3), δ (ppm): 7.55 (d, 2H, J = 7.34Hz), 7.47 (d, 2H, J = 8.26Hz), 7.37-7.41 (m, 4H), 7.34 (d, 2 H,J=5.23Hz),7.30(d,1H,J=7.34Hz),6.55(brs,1H),4.94(s,1H),4.03-4.14(m,4H),1.20(t,6H,J=7.10Hz), 13 C-NMR (100MHz, CDCl3), δ (ppm): 167.1 (2C), 146.0, 141.2, 139.2, 133.7 (2C), 128. 7(2C),128.6(2C),127.0(3C),126.8(2C),108.5(2C),60.1(2C),37.3,14.2(2C).

[0077] Intermediate 13a, 1 H-NMR (400MHz, CDCl3), δ (ppm): 7.27-7.36 (m, 4H), 6.89-6.96 (t, 2H, J = 8.78 Hz),6.33(brs,1H),4.89(s,1H),4.01-4.15(m,4H),1.19(t,6H,J=6.96Hz), 13 C-NMR (100MHz, CDCl3), δ (ppm): 166.9 (2C), 133.4 (2C), 129.8 (2C), 114.8 (2C), 114.6 (2C), 108.7 (2C), 60.1 (2C), 37.0, 14.2 (2C).

[0078] Intermediate 14a, 1 H-NMR (400MHz, CDCl3), δ (ppm): 7.29 (d, 2H, J = 5.28Hz), 7.23 (d, 2H, J = 8.53Hz), 6.77 (d, 2H, J = 8.58Hz), 6.58 (brs,1H),4.83(s,1H),4.02-4.12(m,4H),3.94-4.00(m,2H),1.37(t,3H,J=7.01Hz),1.19(t,6H,J=7.13Hz), 13C-NMR (100MHz, CDCl3), δ (ppm): 167.3 (2C), 157.5, 139.4, 133.4 (2C), 129.3 (2C), 113.9 (2C), 108.7 (2C), 63.3, 60.0 (2C), 36.7, 14.9, 14.2 (2C).

[0079] Intermediate 15a, 1 H-NMR (400MHz, CDCl3), δ (ppm): 7.34-7.43 (m, 4H), 7.28-7.33 (m, 3H), 7.24 (s, 1H), 6.86 (d, 2H, J=8.72Hz),6.21(brs,1H),5.00(s,2H),4.85(s,1H),4.03-4.13(m,4H),1.19(t,6H,J=7.11Hz), 13 C-NMR (100MHz, CDCl3), δ (ppm): 167.1 (2C), 157.4, 139.7, 137.2, 133.2 (2C), 129.3 (2C), 128.5 (2C), 127.9, 127.6 (2C), 114.2 (2C), 109.0 (2C), 70.0, 60.0 (2C), 36.7, 14.2 (2C). Intermediate 16a, 1 H-NMR (600MHz, CDCl3), δ (ppm): 7.39 (d, 1H, J = 2.02Hz), 7.35 (d, 2H, J = 5.13Hz), 7.31 (d, 1H, J = 8.21H z),7.20(dd,1H,J=2.00Hz),6.42(brs,1H),4.88(s,1H),4.04-4.15(m,4H),1.21(t,6H,J=7.15Hz), 13 C-NMR (100MHz, CDCl3), δ (ppm): 166.6 (2C), 147.1, 133.8 (2C), 132.0, 130.4, 130.3, 129.8, 127.9, 108.0 (2C), 60.3 (2C), 37.2, 14.2 (2C).

[0080] Intermediate 17a, 1H-NMR (400 MHz, CDCl3), δ (ppm): 7.38(d,1H,J=7.92Hz),7.34(d,2H,J=5.24Hz),6.95(d,1H,J=1.85Hz),6.78(dd,1H,J=1.85Hz),6.17(brs,1H),4.87(s,1H),4.06-4.12(m,6H),1.44(t,3H,J=6.98Hz),1.20(t,6H,J=7.12Hz), 13 C-NMR (100 MHz, CDCl3), δ (ppm): 166.8(2C),154.8,147.5,133.4(2C),132.7(2C),121.6(2C),113.8,108.5,64.7,60.2(2C),37.5,14.7,14.3(2C).

[0081] Intermediate 18a, 1 H-NMR (400 MHz, CDCl3), δ (ppm): 7.68(d,1H,J=2.06Hz),7.33(d,2H,J=5.33Hz),7.29(dd,1H,J=2.06Hz),6.70(d,1H,J=8.59Hz),6.25(brs,1H),4.81(s,1H),4.02-4.17(m,4H),3.83(s,3H),1.21(t,6H,J=7.04Hz), 13 C-NMR (100 MHz, CDCl3), δ (ppm): 166.9(2C),156.6,141.5,139.2,133.4(3C),129.5,110.3,108.6,85.7,60.1(2C),56.3,36.5,14.2(2C).

[0082] Intermediate 19a, 1 H-NMR (600 MHz, CDCl3), δ (ppm): 7.50(d,2H,J=8.25Hz),7.46(d,2H,J=8.25Hz),7.36(d,2H,J=5.30Hz),6.53(brs,1H),4.98(s,1H),4.04-4.12(m,4H),1.19(t,6H,J=

[0083] 7.15Hz), 13C-NMR (100MHz, CDCl3), δ (ppm): 166. (2C), 150.5, 133.8 (3C), 128.7 (3C), 125.0 (2C), 108.2 (2C), 60.2 (2C), 37.7, 14.2 (2C).

[0084] Intermediate 20a, 1 H-NMR (500MHz, CDCl3), δ (ppm): 7.82 (d, 2H, J = 8.55Hz), 7.56 (d, 2H, J = 8.36Hz), 7.39 (d, 2H, J = 5.28Hz),6.60(brs,1H),5.03(s,1H),4.04-4.13(m,4H),3.03(s,3H),1.19(t,6H,J=7.10Hz), 13 C-NMR (100MHz, CDCl3), δ (ppm): 166.6 (2C), 152.9, 138.3, 134.1 (2C), 129.4 (2C), 127.2 (2C), 107.7 (2C), 60.3 (2C), 44.6, 38.0, 14.2 (2C).

[0085] Intermediate 21a, 1 H-NMR (400MHz, CDCl3), δ (ppm): 7.33 (d, 2H, J = 5.24Hz), 7.23 (d, 2H, J = 7.97Hz), 7. 04(d,2H,J=8.01Hz),6.12(brs,1H),4.87(s,1H),4.02-4.14(m,4H),2.53(t,2H,J=

[0086] 7.76Hz),1.53-1.60(m,4H),1.19(t,6H,J=7.20Hz),0.87(t,3H,J=7.15Hz), 13 C-NMR (100MHz, CDCl3), δ (ppm): 166.6 (2C), 150.5, 133.7 (2C), 128.7 (2C), 125.0, 124.9,124.4,108.2(2C),60.2(2C),37.7,35.0,31.5,30.1,22.7,14.2(2C),14.1.

[0087] Intermediate 22a, 1H-NMR (600MHz, DMSO-d6), δ (ppm): 7.33 (d, 2H, J = 5.45Hz), 7.07 (d, 2H, J = 8.10Hz), 7.03 (d, 2H, J = 8.10Hz), 4.67 ( s,1H),3.93-4.04(m,4H),1.48-1.58(m,2H),1.14-1.28(m,2H),1.12(t,6H,J=7.15Hz),0.87(t,3H,J=7.15Hz), 13 C-NMR (100MHz, DMSO-d6), δ (ppm): 166.9 (2C), 145.1, 135.3 (2C), 128.3 (2C) ,127.9(2C),106.4(2C),61.8,59.8(2C),37.4,36.9,24.5,14.6(2C),14.2.

[0088] Intermediate 23a, 1 H-NMR (600MHz, DMSO-d6), δ (ppm): 7.34 (d, 2H, J = 5.16Hz), 7.08 (d, 2H, J = 7.97Hz), 7.03 (d, 2H, J = 7.97Hz), 4.68 ( s,1H),3.94-4.04(m,4H),1.47-1.52(m,2H),1.14-1.39(m,4H),1.12(t,6H,J=7.14Hz),0.87(t,3H,J=7.30Hz), 13 C-NMR (100MHz, DMSO-d6), δ (ppm): 166.9 (2C), 145.0, 140.6, 135.3 (2C), 128.3 ( 2C),127.9(2C),106.4(2C),59.7(2C),36.9,34.9,33.6,22.3,14.6(2C),14.2.

[0089] General Synthetic Methods for Compounds 1-50

[0090] In a 50 mL round-bottom flask, intermediate 1-23a (0.17 mmol) was dissolved in anhydrous dichloromethane (7 mL), followed by the addition of triethylamine (0.85 mmol), and then slowly added dropwise with a small amount of anhydrous DCM solution of different substituted acyl chlorides (0.26 mmol). The mixture was stirred overnight at room temperature. The reaction was monitored by TLC, and 25 mL of water was added to quench the reaction. The mixture was stirred for 10 min at room temperature. The mixture was extracted with ethyl acetate (3 × 15 mL), and the organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a brown crude product. The crude product was purified by silica gel column chromatography in a PE:EA = 2:1 elution system to give final product 1-51, a white solid, in 35-40% yield.

[0091] The following are the NMR results for compounds 1-50.

[0092] Compound 1, 1 H-NMR (600MHz, DMSO-d6), δ (ppm): 7.96 (s, 2H), 7.77 (d, 2H, J = 8.27Hz), 7.56 (d, 2H, J = 8.27Hz), 7.06(s,2H),4.84(s,1H),3.99-4.08(m,4H),3.82(s,6H),3.77(s,3H),1.09(t,6H,J=7.27Hz), 13 C-NMR (100MHz, CDCl3), δ (ppm): 167.4, 165.3 (2C), 153.5 (2C), 153.4, 149.1, 132.5, 132.2 (2C), 132.0 (2C), 12 9.3(2C),128.4,125.9,114.1,106.9(2C),106.8,61.1(3C),56.5(2C),39.2,14.1(2C).HRMS(ESI+)m / z:calcd for C 28 H 28 N₂O₈([M+H)) + ),521.1918; found,521.1900; HPLC t R = 12.612 min (96.63% purity).

[0093] Compound 2, 1H-NMR (600 MHz, DMSO-d6), δ (ppm): 7.91(s,2H),7.22(d,2H,J=8.74Hz),7.04(s,2H),6.84(d,2H,J=8.77Hz),4.71(s,1H),4.00-4.08(m,4H),3.82(s,6H),3.77(s,3H),3.71(s,3H),1.11(t,6H,J=7.09Hz), 13 C-NMR (150 MHz, DMSO-d6), δ (ppm): 167.6, 165.6(2C), 158.6, 153.3(3C), 141.2, 136.4, 131.2, 129.9(2C), 127.0, 114.5, 114.0(2C), 107.3(3C), 60.8(4C), 56.7(2C), 37.9, 14.4(2C). HRMS (ESI+) m / z: calcd for C 28 H 31 NO9([M+Na] + ), 548.1891; found, 548.1876;

[0094] HPLC t R =14.741 min (100.00% purity).

[0095] Compound 3, 1 H-NMR (400 MHz, CDCl3), δ (ppm): 8.11(s,2H),7.23(d,2H,J=8.08Hz),7.09(d,2H,J=7.95Hz),6.90(s,2H),4.91(s,1H),4.07-4.017(m,4H),3.94(s,3H),3.89(s,6H),2.30(s,3H),1.20(t,6H,J=7.10Hz), 13 C-NMR (100 MHz, CDCl3), δ (ppm): 167.6, 165.8(2C), 153.4(2C), 142.0, 141.0, 136.7, 131.3(2C), 129.0(2C), 128.3(2C), 126.5, 115.6, 106.8(3C), 61.1, 60.8(2C), 56.4(2C), 38.4, 21.1, 14.4(2C). HRMS (ESI+) m / z: calcd for C 28 H 31 NO8([M+H] +),510.2122; found,510.2108; HPLC t R = 15.186 min (97.91% purity).

[0096] Compound 4, 1 H-NMR (600MHz, CDCl3), δ (ppm): 8.13 (s, 2H), 7.41 (d, 2H, J = 8.21Hz), 7.22 (d, 2H, J = 8.21Hz), 6 .88(s,2H),4.92(s,1H),4.08-4.17(m,4H),3.94(s,3H),3.89(s,6H),1.20(t,6H,J=7.15Hz), 13 C-NMR (100MHz, CDCl3), δ (ppm): 167.5, 165.5 (2C), 153.4 (2C), 143.0, 142.1, 131.6 (2C), 131.4 (3C), 130. 2(3C),126.2,121.0,114.8,106.8(2C),61.1,60.9(2C),56.4(2C),38.5,14.1(2C).HRMS(ESI+)m / z:calcd for C 27 H 28 BrNO8([M+H] + ),574.1071; found,574.1060; HPLC t R = 19.486 min (95.20% purity).

[0097] Compound 5, 1 H-NMR (600MHz, DMSO-d6), δ (ppm): 7.92 (s, 2H), 7.01 (s, 2H), 6.77-6.81 (m, 3H), 5.99 (s ,2H),4.86(s,1H),4.01-4.09(m,4H),3.82(s,6H),3.77(s,3H),1.11(t,6H,J=7.11Hz), 13 C-NMR (150MHz, DMSO-d6), δ (ppm): 167.4, 165.5 (2C), 153.4 (3C), 147.5, 145.8, 141.3, 132.2, 126.9, 125.4, 12 3.4,121.7,112.3,107.8,107.2(3C),101.2,60.9(2C),60.8,56.7(2C),34.2,14.4(2C).HRMS(ESI+)m / z:calcd for C 28H 29 NO 10 ([M+Na)) + ),562.1684; found,562.1674; HPLC t R = 13.065 min (98.23% purity).

[0098] Compound 6, 1 H-NMR (600MHz, DMSO-d6), δ (ppm): 7.96 (s, 2H), 7.89 (d, 2H, J = 8.33Hz), 7.49 (d, 2H, J = 8.34Hz), 7.06 ( s,2H),4.85(s,1H),3.98-4.08(m,4H),3.83(s,3H),3.82(s,6H),3.77(s,3H),1.09(t,6H,J=7.13Hz), 13 C-NMR (150MHz, DMSO-d6), δ (ppm): 167.6, 166.5, 165.4 (2C), 153.3 (3C), 149.6, 141.3, 132.0, 129.6 (2C), 129.4 ( 2C),128.8,126.9,113.5,107.4(3C),61.0(2C),60.8,56.7(2C),52.6,39.1,14.4(2C).HRMS(ESI+)m / z:calcdfor C 29 H 31 NO 10 ([M+H)) + ),554.2021; found,554.2005; HPLC t R = 13.440 min (95.47% purity).

[0099] Compound 7, 1 H-NMR (400MHz, CDCl3), δ (ppm): 8.13 (s, 2H), 7.61 (d, 2H, J = 8.31Hz), 7.09 (d, 2H, J = 8.31Hz), 6 .87(s,2H),4.90(s,1H),4.06-4.17(m,4H),3.94(s,3H),3.89(s,6H),1.20(t,6H,J=7.11Hz), 13C-NMR (150MHz, DMSO-d6), δ (ppm): 167.6, 165.4 (2C), 153.3 (2C), 147.6, 144.2, 141.2, 138.4 (2C), 137.4 (2C), 131.3(2C),126.9,118.9,107.4(2C),93.6,61.0(2C),60.8,56.8(2C),38.6,14.4(2C).HRMS(ESI+)m / z:calcd for C 27 H 28 INO8([M+H)) + ),622.0932; found,622.0916;

[0100] HPLC t R = 20.515 min (98.14% purity).

[0101] Compound 8, 1 H-NMR (600MHz, DMSO-d6), δ (ppm): 8.42 (d, 1H, J = 2.13Hz), 8.16 (d, 1H, J = 2.13Hz), 7.97 (s, 2H), 7.09(s,2H),4.85(s,1H),4.00-4.11(m,4H),3.83(s,6H),3.78(s,3H),1.10(t,6H,J=7.11Hz), 13 C-NMR (150MHz, DMSO-d6), δ (ppm): 167.6, 165.2 (2C), 153.3 (3C), 148.6, 146.4, 141.5, 141.2, 139.8, 13 2.8,129.5,126.9,111.8,107.5(3C),61.1(2C),60.8,56.7(2C),36.2,14.3(2C).HRMS(ESI+)m / z:calcd for C 26 H 26 Cl2N2O8([M+H)) + ),565.1139; found,565.1123; HPLC t R = 15.196 min (98.92% purity).

[0102] Compound 9, 1H-NMR (500 MHz, CDCl₃), δ (ppm): 8.18(s,2H),7.76-7.80(m,4H),7.50-7.52(m,1H),7.43-7.46(m,2H),6.94(s,2H),5.13(s,1H),4.05-4.13(m,4H),3.94(s,3H),3.88(s,6H),1.18(t,6H,J=7.42Hz), 13 C-NMR (125 MHz, CDCl₃), δ (ppm): 167.6, 165.7(2C), 153.4(2C), 142.0, 141.4, 133.4, 132.6, 131.4, 127.9, 127.8, 127.6, 127.3, 126.8, 126.4, 126.1, 125.8, 115.4(2C), 106.9(3C), 61.1, 60.9(2C), 56.4(2C), 39.0, 14.1(2C). HRMS (ESI+) m / z: calcd for C 31 H 31 NO₈([M+Na] + ), 568.1942; found, 568.1926; HPLC t R =19.345 min (96.57% purity).

[0103] Compound 10, 1 H-NMR (400 MHz, CDCl₃), δ (ppm): 8.11(s,2H),7.01-7.07(m,1H),6.96(s,2H),6.86-6.93(m,1H, Hz),6.78-6.82(m,1H),5.10(s,1H),4.06-4.18(m,4H),3.94(s,3H),3.89(s,6H),3.80(s,3H),1.20(t,6H,J=7.11Hz), 13 C-NMR (125 MHz, CDCl₃), δ (ppm): 167.4, 165.8(2C), 153.4(2C), 142.0, 132.5(2C), 126.5, 118.4, 118.2, 114.5, 114.3, 113.5, 112.5(2C), 107.0(3C), 61.1, 60.7(2C), 56.5, 56.4(2C), 35.8, 14.1(2C). HRMS (ESI+) m / z: calcd for C 28 H 30 FNO₉([M+H] +), 544.1977; found, 544.1959; HPLC t R =15.501 min (99.38% purity).

[0104] Compound 11, 1 H-NMR (600 MHz, DMSO-d6), δ (ppm): 7.97(s,2H),7.64(d,2H,J=7.62Hz),7.59(d,2H,J=8.19Hz),7.45(t,2H,J=7.62Hz),7.41(d,2H,J=8.19Hz),7.35(t,1H,J=7.62Hz),7.07(s,2H),4.82(s,1H),4.01-4.12(m,4H),3.83(s,6H),3.77(s,3H),1.12(t,6H,J=7.09Hz), 13 C-NMR (150 MHz, DMSO-d6), δ (ppm): 167.1, 165.0(2C), 152.3(2C), 142.9, 140.6, 139.7, 138.7, 131.1, 128.9(2C), 128.8(4C), 126.5(5C), 126.4, 113.5, 106.8(2C), 60.3(2C), 60.2, 56.2(2C), 37.9, 13.8(2C). HRMS (ESI+) m / z: calcd for C 33 H 33 NO8([M+Na] + ), 594.2098; found, 594.2088; HPLC t R =17.496 min (97.50% purity).

[0105] Compound 12, 1 H-NMR (400 MHz, CDCl3), δ (ppm): 8.12(s,2H),7.28-7.32(m,2H),6.97(t,2H,J=7.61Hz),6.88(s,2H),4.94(s,1H),4.07-4.18(m,4H),3.94(s,3H),3.89(s,6H),1.20(t,6H,J=7.11Hz), 13C-NMR (150MHz, CDCl3), δ (ppm): 167.5, 165.6 (2C), 153.4 (2C), 142.0, 139.7, 131.4 (2C), 130.0, 129.9, 12 6.2,115.2(2C),115.1(2C),106.8(3C),61.1,60.9(2C),56.4(2C),38.2,14.1(2C).HRMS(ESI+)m / z:calcd for C 27 H 28 FNO8([M+H)) + ),514.1872; found,514.1859; HPLC t R = 15.920 min (97.56% purity).

[0106] Compound 13, 1 H-NMR (600MHz, DMSO-d6), δ (ppm): 7.90 (s, 2H), 7.20 (d, 2H, J = 8.72Hz), 7.04 (s, 2H), 6.82 (d, 2H, J = 8.72Hz), 4.70 (s, 1H),4.00-4.08(m,4H),3.95-3.99(m,2H),3.82(s,6H),3.77(s,3H),1.29(t,3H,J=7.05Hz),1.11(t,6H,J=7.08Hz), 13 C-NMR (100MHz, CDCl3), δ (ppm): 167.6, 165.8 (2C), 158.0, 153.4 (2C), 141.9, 136.0, 131.1 (2C), 129.4 (3C), 126 .5,115.7,114.2(2C),106.8(2C),63.4,61.1,60.8(2C),56.4(2C),38.0,14.9,14.2(2C).HRMS(ESI+)m / z:calcd for C 29 H 33 NO9([M+Na)) + ),562.2048; found,562.2035; HPLC t R = 16.967 min (98.28% purity).

[0107] Compound 14, 1H-NMR (600 MHz, CDCl3), δ (ppm): 8.10(s,2H),7.30-7.43(m,5H),7.24(s,1H),7.04(s,2H),6.89(d,4H,J=8.15Hz),5.02(s,2H),4.90(s,1H),4.06-4.20(m,4H),3.94(s,3H),3.88(s,6H),1.20(t,6H,J=7.15Hz), 13 C-NMR (150 MHz, DMSO-d6), δ (ppm): 167.0, 165.0(2C), 157.2, 152.7(2C), 140.6, 137.0, 136.1, 130.6, 129.4(2C), 128.3(3C), 127.7, 127.6(3C), 126.5, 114.3(2C), 113.9, 106.7(2C), 69.0, 60.3(2C), 60.2, 56.2(2C), 37.3, 13.8(2C). HRMS (ESI+) m / z: calcd for C 34 H 35 NO9([M+Na] + ), 624.2204; found, 624.2195; HPLC t R =17.183 min (97.65% purity).

[0108] Compound 15, 1 H-NMR (500 MHz, CDCl3), δ (ppm): 8.14(s,2H),7.34-7.38(m,3H),7.21-7.23(m,1H),6.88(s,2H),4.92(s,1H),4.10-4.18(m,4H),3.95(s,3H),3.90(s,6H),1.22(t,6H,J=7.02Hz), 13 C-NMR (100 MHz, CDCl3), δ (ppm): 167.5, 165.4(2C), 153.5(2C), 144.2, 132.4, 131.9(2C), 131.1, 130.3, 130.2, 128.1, 126.1, 114.3(2C), 106.8(3C), 61.1, 61.0(2C), 56.4(2C), 38.3, 14.1(2C). HRMS (ESI+) m / z: calcd for C 27 H 27 Cl2NO8([M+H] +), 564.1186; found, 564.1169; HPLC t R =

[0109] 21.063 min (99.08% purity).

[0110] Compound 16, 1 H-NMR (600 MHz, DMSO-d6), δ (ppm): 7.93(s,2H),7.48(d,1H,J=8.10Hz),7.03(s,2H),6.97(d,1H,J=2.16Hz),6.84(m,1H,J=2.15Hz),4.76(s,1H),4.03-4.11(m,6H),3.82(s,6H),3.78(s,3H),1.34(t,3H,J=6.91Hz),1.11(t,6H,J=7.24Hz), 13 C-NMR (150 MHz, DMSO-d6), δ (ppm): 167.0, 165.0(2C), 154.1, 152.7(3C), 144.9, 140.7, 13.9(2C), 132.6, 131.2, 126.3, 121.8, 114.0, 113.0, 109.5, 106.8(3C), 64.3, 60.4(2C), 60.2, 56.1(2C), 38.2, 14.4. HRMS (ESI+) m / z: calcd for C 29 H 32 BrNO9([M+H] + ), 618.1333; found, 618.1319; HPLC t R =17.176 min (95.00% purity).

[0111] Compound 17, 1 H-NMR (500 MHz, CDCl3), δ (ppm): 8.10(s,2H),7.66(d,1H,J=2.09Hz),7.33(dd,1H,J=2.14Hz),6.90(s,2H),6.74(d,1H,J=8.43Hz),4.85(s,1H),4.09-4.18(m,4H),3.94(s,3H),3.91(s,6H),3.85(s,3H),1.22(t,6H,J=7.05Hz), 13C-NMR (125MHz, CDCl3), δ (ppm): 167.6, 165.6 (2C), 157.1, 153.4 (2C), 142.0, 139.1, 138.3, 131.5 (2C), 129.9 ( 2C),126.3,115.0,110.5,106.8(3C),61.1,60.9(2C),56.5(2C),56.3,37.6,14.2(2C).HRMS(ESI+)m / z:calcd for C 28 H 30 INO9([M+Na)) + ),674.0857; found,674.0837; HPLCt R = 16.208 min (96.57% purity).

[0112] Compound 18, 1 H-NMR (400MHz, CDCl3), δ (ppm): 8.17 (s, 2H), 7.55 (d, 2H, J = 8.17Hz), 7.46 (d, 2H, J = 8.17Hz), 6 .89(s,2H),5.03(s,1H),4.08-4.18(m,4H),3.95(s,3H),3.89(s,6H),1.20(t,6H,J=7.11Hz), 13 C-NMR (125MHz, CDCl3), δ (ppm): 167.5, 165.4 (2C), 153.5 (2C), 147.7, 142.2, 131.8 (2C), 128.8 (3C), 1 26.1,125.3(2C),114.6,106.9(4C),61.1,61.0(2C),56.5(2C),38.9,14.1(2C).HRMS(ESI+)m / z:calcd for C 28 H 28 F3NO8([M+H) + ),564.1840; found,564.1833; HPLC t R = 18.793 min (96.44% purity).

[0113] Compound 19, 1H-NMR (400 MHz, CDCl3), δ (ppm): 8.18 (s, 2H), 7.87 (d, 2H, J=8.30 Hz), 7.55 (d, 2H, J=8.30 Hz), 6.88 (s, 2H), 5.06 (s, 1H), 4.06-4.19 (m, 4H), 3.95 (s, 3H), 3.90 (s, 6H), 3.05 (s, 3H), 1.20 (t, 6H, J=7.15 Hz), 13 C-NMR (150 MHz, CDCl3), δ (ppm): 167.4, 165.3 (2C), 153.5 (2C), 150.0, 142.2, 139.2, 132.0 (2C), 129.5 (3C), 127.5 (2C), 125.9, 114.1, 106.9 (2C), 61.1 (3C), 56.5 (2C), 44.5, 39.0, 14.1 (2C). HRMS (ESI+) m / z: calcd for C 28 H 31 NO 10 S([M+H] + ), 574.1741; found, 574.1732; HPLC t R =10.389 min (97.96% purity).

[0114] Compound 20, 1 H-NMR (600 MHz, DMSO-d6), δ (ppm): 7.91 (s, 2H), 7.19 (d, 2H, J=8.15 Hz), 7.09 (d, 2H, J=8.15 Hz), 7.02 (s, 2H), 4.72 (s, 1H), 3.98-4.07 (m, 4H), 3.81 (s, 6H), 3.75 (s, 3H), 1.49-1.54 (m, 2H), 1.14-1.35 (m, 6H), 1.08 (t, 6H, J=7.15 Hz), 0.83 (t, 3H, J=7.03 Hz), 13 C-NMR (125 MHz, CDCl3), δ (ppm): 167.6, 165.8 (2C), 153.4 (2C), 141.7, 141.0, 131.3 (2C), 128.3 (3C), 128.2 (2C), 126.5, 115.6, 106.8 (3C), 61.1, 60.8 (2C), 56.4 (2C), 38.4, 35.6, 31.6, 31.0, 22.5, 14.1 (2C), 14.0. HRMS (ESI+) m / z: calcd for C 32 H 39NO8([M+Na] + ), 588.2568; found, 588.2550; HPLC t R =20.996 min (95.46% purity).

[0115] Compound 21, 1 H-NMR (400 MHz, CDCl3), δ (ppm): 8.11(s, 2H), 7.24(d, 2H, J=8.15 Hz), 7.09(d, 2H, J=8.15 Hz), 6.90(s, 2H), 4.92(s, 1H), 4.07-4.18(m, 4H), 3.94(s, 3H), 3.89(s, 6H), 2.54(t, 2H, J=7.25 Hz), 1.58-1.64(m, 2H), 1.20(t, 6H, J=7.08 Hz), 0.87(t, 3H, J=7.27 Hz), 13 C-NMR (125 MHz, CDCl3), δ (ppm): 167.6, 165.8(2C), 153.4, 141.9, 141.4, 141.1, 131.3, 128.4(3C), 128.2(3C), 126.5, 115.6, 106.8(3C), 61.1, 60.8(2C), 56.4(2C), 38.4, 37.7, 24.4, 14.1(2C), 13.9. HRMS (ESI+) m / z: calcd for C 30 H 35 NO8([M+H] + ), 538.2435; found, 538.2433; HPLC t R =18.097 min (97.56% purity).

[0116] Compound 22, 1 H-NMR (600 MHz, CDCl3), δ (ppm): 8.11(s, 2H), 7.23(d, 2H, J=8.09 Hz), 7.09(d, 2H, J=8.09 Hz), 6.89(s, 2H), 4.92(s, 1H), 4.07-4.19(m, 4H), 3.94(s, 3H), 3.89(s, 6H), 2.52-2.59(m, 2H), 1.23-1.45(m, 4H), 1.20(t, 6H, J=7.17 Hz), 0.91(t, 3H, J=7.36 Hz), 13C-NMR (150MHz, CDCl3), δ (ppm): 167.6, 165.8 (2C), 153.4 (2C), 142.0, 141.7, 141.0, 131.3 (2C), 128.4 (2C), 128.2 (2C), 126.5,115.6(2C),106.8(2C),61.1,60.8(2C),56.4(2C),38.4,35.3,33.5,22.4,14.1(2C),14.0.HRMS(ESI+)m / z:calcd for C 31 H 37 NO8([M+Na)) + ),574.2411; found,574.2400;

[0117] HPLC t R = 19.629 min (95.19% purity).

[0118] Compound 23, 1 H-NMR (600MHz, CDCl3), δ (ppm): 8.15 (s, 2H), 7.88 (d, 2H, J = 8.42Hz), 7.63-7.67 (m, 3H), 7.55- 7.57(m,4H),4.89(s,1H),5.06(s,1H),4.07-4.16(m,4H),3.05(s,3H),1.19(t,6H,J=7.11Hz), 13 C-NMR (150MHz, CDCl3), δ (ppm): 167.9, 165.3 (2C), 149.9, 139.1, 133.0, 131.9, 131.3, 129.5 (3C),129.1(4C),127.5(3C),114.3,61.1(2C),44.5,39.0,14.1(2C).HRMS(ESI+)m / z:calcd for C 25 H 25 NO7S([M+H)) + ),484.1424; found,484.1383; HPLC t R = 11.029 min (96.00% purity).

[0119] Compound 24, 1H-NMR (400 MHz, CDCl3), δ (ppm): 8.00 (s, 2H), 7.87 (d, 2H, J=8.37 Hz), 7.52 (d, 2H, J=8.37 Hz), 7.45-7.49 (m, 1H), 7.33-7.36 (m, 3H), 5.03 (s, 1H), 4.04-4.14 (m, 4H), 3.04 (s, 3H), 2.39 (s, 3H), 1.18 (t, 6H, J=7.14 Hz), 13 C-NMR (150 MHz, CDCl3), δ (ppm): 168.2, 165.2 (2C), 149.7, 139.1, 131.4, 131.3, 130.9, 129.6 (4C), 127.4 (5C), 126.4, 114.7, 61.1 (2C), 44.5, 39.1, 19.5, 14.1 (2C). HRMS (ESI+) m / z: calcd for C 26 H 27 NO7S ([M+H] + ), 498.1580; found, 498.1540; HPLC t R =13.703 min (95.14% purity).

[0120] Compound 25, 1 H-NMR (600 MHz, CDCl3), δ (ppm): 8.15 (s, 2H), 7.88 (d, 2H, J=8.26 Hz), 7.56 (d, 4H, J=7.52 Hz), 7.35 (d, 2H, J=8.07 Hz), 5.06 (s, 1H), 4.08-4.16 (m, 4H), 3.05 (s, 3H), 2.47 (s, 3H), 1.20 (t, 6H, J=7.12 Hz), 13 C-NMR (150 MHz, CDCl3), δ (ppm): 167.9, 165.4 (2C), 150.1, 144.0, 139.1, 132.2 (2C), 129.7 (2C), 129.5 (2C), 129.3 (2C), 127.5 (2C), 114.0, 61.0 (2C), 44.5, 39.0, 21.7, 14.1 (2C). HRMS (ESI+) m / z: calcd for C 26 H 27 NO7S ([M+H] + ), 498.1580; found, 498.1534; HPLC t R =13.242 min (98.95% purity).

[0121] Compound 26, 1 H-NMR (400MHz, CDCl3), δ (ppm): 8.15 (s, 2H), 7.88 (d, 2H, J = 8.11Hz), 7.55 (d, 2H, J = 8.19Hz), 7.45 (t, 1H, J = 8 .05Hz),7.15-7.19(m,3H),5.05(s,1H),4.07-4.16(m,4H),3.88(s,3H),3.05(s,3H),1.20(t,6H,J=7.18Hz), 13 C-NMR (125MHz, CDCl3), δ (ppm): 167.7, 165.3 (2C), 160.1, 149.9, 139.1, 132.6, 131.9, 130.1, 129.5 (3C ),127.5(3C),120.9,118.8,114.4,114.3,61.1(2C),55.6,44.5,39.0,14.1(2C).HRMS(ESI+)m / z:calcd for C 26 H 27 NO8S([M+H)) + ),514.1529; found,514.1490; HPLC t R = 11.500 min (97.39% purity).

[0122] Compound 27, 1 H-NMR (400MHz, CDCl3), δ (ppm): 8.16 (s, 2H), 7.88 (d, 2H, J = 8.31Hz), 7.66 (d, 2H, J = 8.78Hz), 7.56 (d, 2H, J = 8.3 1Hz),7.03(d,2H,J=8.78Hz),5.06(s,1H),4.07-4.17(m,4H),3.91(s,3H),3.05(s,3H),1.20(t,6H,J=7.15Hz), 13 C-NMR (100MHz, CDCl3), δ (ppm):

[0123] 167.5,165.5(2C),163.5,150.2,139.1,132.5(2C),131.7(2C),129.5(2C),127.5(3C), 123.1,114.4(2C),113.7,61.0(2C),55.6,44.5,39.0,14.1(2C).HRMS(ESI+)m / z:calcd for C26 H 27 NO8S([M+H] + ), 514.1529; found, 514.1486; HPLC t R =11.626 min (98.25% purity).

[0124] Compound 28, 1 H-NMR (500 MHz, CDCl3), δ (ppm): 8.17 (s, 2H), 7.88 (d, 2H, J=8.38 Hz), 7.57 (d, 2H, J=8.38 Hz), 7.27 (s, 1H), 7.21-7.25 (m, 1H), 6.97 (d, 2H, J=8.36 Hz), 5.07 (s, 1H), 4.08-4.17 (m, 4H), 3.98 (s, 3H), 3.95 (s, 3H), 3.05 (s, 3H), 1.20 (t, 6H, J=7.15 Hz), 13 C-NMR (125 MHz, CDCl3), δ (ppm): 167.5, 165.4 (2C), 153.3, 150.2, 149.6, 139.1, 132.5 (2C), 129.5 (2C), 127.7, 127.5 (2C), 123.2 (2C), 113.8, 112.5, 110.5, 61.0 (2C), 56.2 (2C), 44.5, 39.0, 14.2 (2C). HRMS (ESI+) m / z: calcd for C 27 H 29 NO9S([M+H] + ), 544.1635; found, 544.1588; HPLC t R =9.810 min (88.72% purity).

[0125] Compound 29, 1 H-NMR (400 MHz, CDCl3), δ (ppm): 8.09 (s, 2H), 7.89 (d, 2H, J=8.35 Hz), 7.84 (d, 2H, J=8.35 Hz), 7.76 (d, 2H, J=8.18 Hz), 7.55 (d, 2H, J=8.35 Hz), 5.06 (s, 1H), 4.07-4.17 (m, 4H), 3.05 (s, 3H), 1.19 (t, 6H, J=7.10 Hz), 13C-NMR (125 MHz, CDCl3), δ (ppm): 166.5, 165.1 (2C), 149.5, 139.3, 131.1, 129.6 (4C), 129.4 (4C), 127.6 (4C), 126.2, 115.2, 61.2 (2C), 44.5, 39.1, 14.1 (2C). HRMS (ESI+) m / z: calcd for C 26 H 24 F3NO7S ([M+H] + ), 552.1298; found, 552.1250; HPLC t R =

[0126] 14.396 min (95.41% purity).

[0127] Compound 30, 1 H-NMR (400 MHz, CDCl3), δ (ppm): 8.12 (s, 2H), 7.88 (d, 2H, J=8.35 Hz), 7.68-7.72 (m, 2H), 7.55 (d, 2H, J=8.35 Hz), 7.24 (d, 2H, J=7.44 Hz), 5.06 (s, 1H), 4.07-4.17 (m, 4H), 3.05 (s, 3H), 1.20 (t, 6H, J=7.10 Hz), 13 C-NMR (100 MHz, CDCl3), δ (ppm): 166.8, 165.2 (2C), 149.8, 139.2, 131.9 (2C), 131.7 (3C), 129.5 (3C), 127.5 (3C), 116.6, 116.4, 114.5, 61.1 (2C), 44.5, 39.0, 14.1 (2C). HRMS (ESI+) m / z: calcd for C 25 H 24 FNO7S ([M+H] + ), 502.1330; found, 502.1285; HPLC t R =11.600 min (96.56% purity).

[0128] Compound 31, 1H-NMR (400 MHz, CDCl3), δ (ppm): 8.10 (s, 2H), 7.88 (d, 2H, J=8.33 Hz), 7.61 (d, 2H, J=8.52 Hz), 7.53-7.56 (m, 4H), 5.06 (s, 1H), 4.07-4.17 (m, 4H), 3.05 (s, 3H), 1.20 (t, 6H, J=7.06 Hz), 13 C-NMR (125 MHz, CDCl3), δ (ppm): 167.0, 166.2 (2C), 149.7, 139.2, 132.5 (3C), 131.5, 130.6 (3C), 130.2, 129.5 (2C), 128.0, 127.5 (2C), 114.7, 61.2 (2C), 44.5, 39.0, 14.1 (2C). HRMS (ESI+) m / z: calcd for C 25 H 24 ClNO7S ([M+H] + ), 518.1034; found, 518.0987; HPLC t R =13.905 min (98.35% purity).

[0129] Compound 32, 1 H-NMR (400 MHz, CDCl3), δ (ppm): 8.10 (s, 2H), 7.88 (d, 2H, J=8.20 Hz), 7.71 (d, 2H, J=8.52 Hz), 7.53-7.56 (m, 4H), 5.06 (s, 1H), 4.07-4.17 (m, 4H), 3.05 (s, 3H), 1.20 (t, 6H, J=7.12 Hz), 13 C-NMR (125 MHz, CDCl3), δ (ppm): 167.0, 165.2 (2C), 149.7, 139.3, 132.5 (3C), 131.5, 130.6 (3C), 129.5 (3C), 127.5 (3C), 114.7, 61.2 (2C), 44.5, 39.0, 14.1 (2C). HRMS (ESI+) m / z: calcd for C 25 H 24 BrNO7S ([M+H] + ), 562.0529; found, 562.0481; HPLC t R =14.658 min (97.92% purity). Compound 33, 1H-NMR (600 MHz, CDCl3), δ (ppm): 8.07-8.08 (m, 2H), 7.96-7.98 (m, 1H), 7.91 (d, 3H, J=8.20 Hz), 7.55-7.65 (m, 7H), 5.05 (s, 1H), 4.02-4.10 (m, 4H), 3.05 (s, 3H), 1.14 (t, 6H, J=7.18 Hz), 13 C-NMR (150 MHz, CDCl3), δ (ppm): 167.7, 165.2 (2C), 149.7, 139.1, 133.7, 132.2, 129.6 (4C), 127.5 (3C), 129.0, 128.2 (2C), 127.2 (2C), 126.6, 124.8, 124.3, 114.7, 61.0 (2C), 44.5, 39.1, 14.0 (2C). HRMS (ESI+) m / z: calcd for C 29 H 27 NO7S ([M+H] + ), 534.1580; found, 534.1527; HPLC t R =14.407 min (97.56% purity).

[0130] Compound 34, 1 H-NMR (500 MHz, CDCl3), δ (ppm): 8.21 (s, 2H), 7.95-8.01 (m, 3H), 7.90-7.92 (m, 3H), 7.66-7.69 (m, 2H), 7.59-7.64 (m, 3H), 5.09 (s, 1H), 4.06-4.15 (m, 4H), 3.06 (s, 3H), 1.18 (t, 6H, J=7.17 Hz), 13 C-NMR (125 MHz, CDCl3), δ (ppm): 168.0, 165.3 (2C), 150.0, 139.2, 132.4, 132.0 (2C), 130.6 (2C), 129.6 (3C), 129.1 (2C), 128.9, 128.0, 127.6, 127.5 (2C), 124.5 (2C), 114.3, 61.1 (2C), 44.5, 39.1, 14.1 (2C). HRMS (ESI+) m / z: calcd for C 29 H 27 NO7S ([M+H] + ), 534.1580;

[0131] found, 534.1535; HPLC tR =15.321min (96.17% purity).

[0132] Compound 35, 1 H-NMR (600MHz, CDCl3), δ (ppm): 8.44(s,2H), 7.85(d,2H,J=8.44Hz), 7.72-7.73(m,1H), 7.55(d,1H,J=8.39Hz), 7.44-7.46(dd,2H), 6.68-6.69(m,1H), 5.06(s,1H), 4.11-4.19(m,4H), 3.03(s,3H), 1.23(t,6H,J=7.09Hz), 13 C-NMR (150MHz, CDCl3), δ (ppm): 165.4(2C), 155.9, 149.7, 147.2, 145.2, 139.1, 131.6, 129.6, 127.5(3C), 122.2, 114.7(2C), 112.8(2C), 61.1(2C), 44.5, 38.9, 14.1(2C). HRMS(ESI+) m / z: calcd for C 23 H 23 NO8S([M+H] + ), 474.1216; found, 474.1170; HPLC t R =8.937min (95.75% purity).

[0133] Compound 36, 1 H-NMR (500MHz, CDCl3), δ (ppm): 8.32(s,2H), 7.87(d,2H,J=8.27Hz), 7.78(d,1H,J=4.96Hz), 7.66(d,2H,J=3.89Hz), 7.55(d,2H,J=8.36Hz), 7.19-7.25(m,1H), 5.08(s,1H), 4.10-4.19(m,4H), 3.04(s,3H), 1.22(t,6H,J=7.06Hz), 13 C-NMR (125MHz, CDCl3), δ (ppm): 165.3(2C), 161.0, 149.8, 139.2, 134.2, 133.5, 132.0(2C), 129.5(2C), 128.1(2C), 127.6(2C), 114.5(2C), 61.1(2C), 44.5, 39.0, 14.1(2C). HRMS(ESI+) m / z: calcd for C 23 H23 NO7S2([M+H] + ), 490.0988; found, 490.0947; HPLC t R =10.925 min (96.64% purity).

[0134] Compound 37, 1 H-NMR (500 MHz, CDCl3), δ (ppm): 8.29 (d, 1H, J=1.87 Hz), 8.14 (s, 2H), 7.73 (d, 1H, J=1.87 Hz), 7.62-7.68 (m, 3H), 7.54-7.59 (m, 2H), 4.96 (s, 1H), 4.09-4.18 (m, 4H), 1.22 (t, 6H, J=7.03 Hz), 13 C-NMR (125 MHz, CDCl3), δ (ppm): 167.8, 165.1 (2C), 147.4 (2C), 139.9, 138.8, 133.0 (2C), 132.2 (2C), 131.2, 130.3, 129.1 (4C), 113.3, 61.3 (2C), 36.0, 14.1 (2C). HRMS (ESI+) m / z: calcd for C 23 H 20 Cl2N2O5([M+H] + ), 475.0821; found, 475.0777; HPLC t R =19.403 min (96.76% purity).

[0135] Compound 38, 1 H-NMR (500 MHz, CDCl3), δ (ppm): 8.26 (d, 1H, J=1.87 Hz), 8.01 (s, 2H), 7.69-7.70 (m, 1H), 7.45-7.49 (m, 1H), 7.31-7.37 (m, 3H), 4.93 (s, 1H), 4.07-4.16 (m, 4H), 2.38 (s, 3H), 1.20 (t, 6H, J=7.06 Hz), 13 C-NMR (125 MHz, CDCl3), δ (ppm): 168.1, 165.0 (2C), 147.4 (2C), 138.8 (2C), 139.8, 131.5 (2C), 131.3 (2C), 130.2, 127.4 (2C), 126.5 (2C), 113.8, 61.2 (2C), 36.1, 19.5, 14.1 (2C). HRMS (ESI+) m / z: calcd for C24 H 22 Cl2N2O5([M+H + ),489.0978; found, 489.0921; HPLC t R =20.906min (95.12% purity).

[0136] Compound 39, 1 H-NMR (500MHz, CDCl3), δ(ppm): 8.29(d,1H,J=2.15Hz), 8.15(s,2H), 7.73(d,1H,J=2.15Hz), 7.55(d,2H,J=8.13Hz), 7.35(d,2H,J=8.13Hz), 4.95(s,1H), 4.10-4.18(m,4H), 2.46(s,3H), 1.22(t,6H,J=7.08Hz), 13 C-NMR (125MHz, CDCl3), δ(ppm): 167.9, 165.1(2C), 147.4, 144.1, 140.0, 138.8, 132.5(2C), 129.8(3C), 129.3(3C), 128.2, 113.0(2C), 61.2, 36.0, 21.7, 14.1(2C). HRMS(ESI+) m / z: calcd for C 24 H 22 Cl2N2O5([M+H + ),489.0978; found, 489.0929; HPLC t R =21.877min (98.24% purity).

[0137] Compound 40, 1 H-NMR (500MHz, CDCl3), δ(ppm): 8.28(d,1H,J=2.17Hz), 8.15(s,2H), 7.72(d,1H,J=2.17Hz), 7.41-7.48(m,1H), 7.11-7.20(m,3H), 4.95(s,1H), 4.10-4.18(m,4H), 3.88(s,3H), 1.22(t,6H,J=7.13Hz), 13C-NMR (125 MHz, CDCl3), δ (ppm): 167.7, 165.0 (2C), 160.1, 147.4 (2C), 139.9, 138.8 (2C), 132.3 (2C), 130.1, 120.9, 119.0 (2C), 114.3 (2C), 113.3, 61.2 (2C), 55.6, 36.0, 14.1 (2C). HRMS (ESI+) m / z: calcd for C 24 H 22 Cl2N2O6 ([M+H + ), 505.0927; found, 505.0870; HPLC t R =18.930 min (98.53% purity).

[0138] Compound 41, 1 H-NMR (500 MHz, CDCl3), δ (ppm): 8.29 (d, 1H, J=2.10 Hz), 8.15 (s, 2H), 7.74 (d, 1H, J=2.09 Hz), 7.65 (d, 2H, J=8.79 Hz), 7.03 (d, 2H, J=8.79 Hz), 4.96 (s, 1H), 4.10-4.19 (m, 4H), 3.91 (s, 3H), 1.22 (t, 6H, J=7.09 Hz), 13 C-NMR (125 MHz, CDCl3), δ (ppm): 167.4, 165.2 (2C), 163.6, 147.3, 140.2, 138.8, 132.8 (2C), 131.7 (3C), 130.2, 122.9, 114.5 (3C), 112.8, 61.2 (2C), 55.7, 36.0, 14.1 (2C). HRMS (ESI+) m / z: calcd for C 24 H 22 Cl2N2O6 ([M+H + ), 505.0927; found, 505.0879; HPLC t R =18.940 min (99.69% purity).

[0139] Compound 42, 1H-NMR (600 MHz, CDCl₃), δ (ppm): 8.30 (d, 1H, J=2.06 Hz), 8.17 (s, 2H), 7.74 (d, 1H, J=2.06 Hz), 7.24-7.26 (m, 1H), 6.97 (d, 2H, J=8.22 Hz), 4.97 (s, 1H), 4.11-4.19 (m, 4H), 3.98 (s, 3H), 3.95 (s, 3H), 1.23 (t, 6H, J=7.09 Hz), 13 C-NMR (150 MHz, CDCl₃), δ (ppm): 167.4, 165.2 (2C), 153.3, 149.5, 147.5, 147.3 (2C), 140.2, 138.8, 132.8 (2C), 130.3, 123.3, 123.0, 112.8, 112.4, 110.5, 61.2 (2C), 56.2 (2C), 36.0, 14.2 (2C). HRMS (ESI+) m / z: calcd for C 25 H 24 Cl₂N₂O₇ ([M+H] + ), 535.1033; found, 535.0975; HPLC t R =17.440 min (96.17% purity). Compound 43, 1 H-NMR (500 MHz, CDCl₃), δ (ppm): 8.28 (d, 1H, J=2.11 Hz), 8.09 (s, 2H), 7.84 (d, 2H, J=8.22 Hz), 7.77 (d, 2H, J=8.22 Hz), 7.72 (d, 2H, J=2.11 Hz), 4.96 (s, 1H), 4.11-4.18 (m, 4H), 1.22 (t, 6H, J=7.16 Hz), 13 C-NMR (125 MHz, CDCl₃), δ (ppm): 166.5, 164.8 (2C), 147.7, 147.4 (2C), 139.5 (2C), 138.8 (2C), 134.7, 131.5, 130.3, 129.4 (3C), 126.3, 126.2, 114.2, 61.4 (2C), 36.1, 14.1 (2C). HRMS (ESI+) m / z: calcd for C 24 H 19 Cl₂F₃N₂O₅ ([M+H] + ), 543.0695; found, 543.0642; HPLC t R =21.626 min (98.11% purity).

[0140] Compound 44, 1 H-NMR (600MHz, CDCl3), δ (ppm): 8.30 (d, 1H, J = 2.20Hz), 8.12 (s, 2H), 7.74 (d, 1H, J = 2.20Hz), 7.67-7.72(m,2H),7.24-7.26(m,2H),4.97(s,1H),4.10-4.20(m,4H),1.22(t,6H,J=7.13Hz), 13 C-NMR (100MHz, CDCl3), δ (ppm): 166.7, 165.0 (2C), 147.6, 147.3 (2C), 139.8, 138.8 (2C), 132.1 (2C) ),131.9,131.8,130.2,116.6,116.4,113.5(2C),61.3(2C),36.0,14.1(2C).HRMS(ESI+)m / z:calcd for C 23 H 19 Cl2FN2O5([M+H) + ),493.0727; found,493.0677; HPLC t R = 18.929 min (96.07% purity).

[0141] Compound 45, 1 H-NMR (500MHz, CDCl3), δ (ppm): 8.28 (s, 1H), 8.10 (s, 2H), 7.72 (s, 1H), 7.60 (d, 2H, J = 8 .42Hz),7.54(d,2H,J=8.42Hz),4.96(s,1H),4.10-4.19(m,4H),1.22(t,6H,J=7.12Hz), 13 C-NMR (125MHz, CDCl3), δ (ppm): 166.8, 164.9 (2C), 147.6, 147.3, 139.7, 139.6, 138.8, 13 1.9(2C),130.5(3C),129.5(4C),113.7,61.3(2C),36.0,14.1(2C).HRMS(ESI+)m / z:calcd for C 23 H 19 Cl3N2O5([M+H) + ),509.0432; found,509.0376; HPLC t R = 21.342 min (98.26% purity). Compound 46,1 H-NMR (600 MHz, CDCl3), δ (ppm): 8.28 (d, 1H, J=2.17 Hz), 8.10 (s, 2H), 7.72 (d, 1H, J=2.17 Hz), 7.71 (d, 2H, J=8.56 Hz), 7.52 (d, 2H, J=8.56 Hz), 4.96 (s, 1H), 4.11-4.19 (m, 4H), 1.22 (t, 6H, J=7.15 Hz), 13 C-NMR (150 MHz, CDCl3), δ (ppm): 166.9, 164.9 (2C), 147.6, 147.3, 139.7, 138.8, 132.5 (3C), 131.9, 130.6 (3C), 130.3, 129.9, 128.1, 113.7, 61.4 (2C), 36.0, 14.1 (2C). HRMS (ESI+) m / z: calcd for C 23 H 19 BrCl2N2O5 ([M+H] + ), 552.9926; found, 552.9901; HPLC t R =22.202 min (98.28% purity).

[0142] Compound 47, 1 H-NMR (500 MHz, CDCl3), δ (ppm): 8.29 (d, 1H, J=2.11 Hz), 8.04-8.12 (m, 2H), 7.98 (d, 1H, J=8.31 Hz), 7.85 (s, 2H), 7.72 (d, 1H, J=2.11 Hz), 7.57-7.70 (m, 4H), 4.94 (s, 1H), 4.05-4.11 (m, 4H), 1.17 (t, 6H, J=7.05 Hz), 13 C-NMR (125 MHz, CDCl3), δ (ppm): 167.7, 164.9 (2C), 147.5, 147.4 (2C), 139.7 (2C), 138.8 (2C), 133.6, 132.3, 131.5, 130.2, 129.0, 128.3, 127.2 (3C), 126.7, 124.9, 124.1, 113.8, 61.2 (2C), 36.1, 14.0 (2C). HRMS (ESI+) m / z: calcd for C 27 H 22 Cl2N2O5 ([M+H] + ), 525.0978; found, 525.0920; HPLC tR =22.831min (99.04% purity). Compound 48, 1 H-NMR (500MHz, CDCl3), δ(ppm): 8.29(d,1H,J=2.11Hz), 8.07-8.09(m,1H), 7.98(d,1H,J=8.31Hz), 7.85(s,2H), 7.72(d,1H,J=2.11Hz), 7.54-7.74(m,4H), 4.94(s,1H), 4.09-4.17(m,4H), 1.17(t,6H,J=7.05Hz), 13 C-NMR (125MHz, CDCl3), δ(ppm): 167.9, 165.1(2C), 147.6, 147.4, 140.0, 138.9(2C), 135.2, 132.4(2C), 130.7, 130.3, 129.2, 129.1, 128.9, 128.2, 128.0, 127.6, 124.5(2C), 113.3, 61.2(2C), 36.1, 14.1(2C). HRMS(ESI+) m / z: calcd for C 27 H 22 Cl2N2O5([M+H] + ), 525.097; found, 525.0927; HPLC t R =23.130min (96.28% purity). Compound 49, 1 H-NMR (500MHz, CDCl3), δ(ppm): 8.44(s,2H), 8.29(d,1H,J=2.17Hz), 7.73(s,1H), 7.70(d,1H,J=2.17Hz), 7.46(d,1H,J=3.52Hz), 6.68-6.69(dd,1H), 4.96(s,1H), 4.13-4.21(m,4H), 1.25(t,6H,J=7.11Hz), 13 C-NMR (125MHz, CDCl3), δ(ppm): 165.1(2C), 155.8, 147.5, 147.3(2C), 145.1, 139.7, 138.7(2C), 132.0(2C), 122.4(2C), 113.7, 112.9, 61.3(2C), 35.9, 14.1(2C). HRMS(ESI+) m / z: calcd for C 21 H 18 Cl2N2O6([M+H] +),465.0614; found,465.0566; HPLC t R = 16.576 min (98.27% purity).

[0143] Compound 50, 1 H-NMR (500MHz, CDCl3), δ (ppm): 8.31 (s, 2H), 8.29 (d, 1H, J = 2.10Hz), 7.79 (d, 1H, J = 4.90Hz), 7.73 (d, 1H, J = 2. 10Hz),7.65(d,1H,J=3.76Hz),7.23(t,1H,J=4.11Hz),4.98(s,1H),4.12-4.21(m,4H),1.24(t,6H,J=7.21Hz), 13 C-NMR (125MHz, CDCl3), δ (ppm): 165.1 (2C), 160.9, 147.6, 147.3, 139.8, 138.7, 134.4, 133.6 ,132.4(2C),130.3,128.2(2C),113.5(2C),61.3(2C),36.0,14.1(2C).HRMS(ESI+)m / z:calcd forC 21 H 18 Cl2N2O5S([M+H) + ),481.0385; found,481.0331; HPLC t R = 17.981 min (99.15% purity).

[0144] SIRT3 enzyme activating activity of compounds 1-50 in Experimental Example 1

[0145] The purpose of this experiment was to detect the in vitro SIRT3 agonist activity of the compounds of the present invention (Table 1).

[0146] All enzymatic reactions were carried out at 37°C for 30 minutes. Each 50 μL reaction mixture contained 25 mM Tris, pH 8.0, 1 mM MgCl2, 0.1 mg / mL BSA, 1.37 mM NaCl, 2.7 mM KCl, SIRT3, and the enzyme substrate. The compounds were diluted with 10% DMSO, and 5 μL of diluent was added to each 50 μL reaction mixture to bring the final concentration of DMSO in all reactions to 1%. Analysis was performed by quantifying the amount of fluorescent product in solution during the enzymatic reaction. Fluorescence analysis was then performed using a SpectraMax M5 microtiter plate reader at excitation wavelengths of 350–360 nm and emission wavelengths of 450–460 nm. IC50 50The values ​​were calculated using nonlinear regression and normalized dose-response fitting with Prism GraphPad software.

[0147] Table 1. Agonistaltic activity of compounds 1-50 against SIRT3

[0148]

[0149]

[0150] Experimental Example 2: Compound 42 inhibited the proliferation of MDA-MB-231 and BT-549 cells.

[0151] BT-549 and MDA-MB-231 cells were digested with trypsin, placed in DMEM medium, centrifuged, and the supernatant was discarded. Culture medium was added and the cells were gently dispersed by pipetting. The cells were then seeded in 6-well plates and cultured in a 5% CO2 incubator at 37°C. Cell growth was observed daily. Once cells entered the logarithmic growth phase, DMSO solution containing compound 42 (experimental group) and DMSO (blank control group) were added, and culture continued. When obvious clones appeared, the culture medium was aspirated, the cells were washed twice with PBS, and then fixed with 4% polymethanol. After 15 minutes, the supernatant was gently aspirated, and the cells were stained with crystal violet. After 30 minutes, the staining solution was washed off, and cell clones were photographed. The colony formation rate was calculated by counting clones with more than 10 cells per well.

[0152] Next, compound 42, with the best agonistic activity, was selected for antiproliferative activity studies. First, we tested the antiproliferative capacity of compound 42 against triple-negative breast cancer cells. The results showed that compound 42 had an antiproliferative effect on MDA-MB-231 cells (6hIC). 50 =60.76±1.784μM; 12h IC 50 =11.16±1.047μM; 24h IC 50 =1.61±0.207μM; 36hIC 50 =0.94±0.270μM; 48h IC 50 =0.76±0.121μM) and BT-549 cells (6h IC50) 50 =4.272±0.631μM; 12h IC 50 =1.81±0.259μM; 24h IC 50 =1.95±0.289μM; 36h IC 50 =1.47±0.168μM; 48hIC 50 Compound 42 (0.93 ± 0.030 μM) exhibited good anti-proliferative activity. Experimental results showed that BT-549 cells were relatively sensitive to compound 42 (0.93 ± 0.030 μM). Figure 1 ).

[0153] Dissolve the prepared 2% low-soluble agarose solution in a 60°C water bath, then quickly add 50 μL / well to a 96-well plate and allow it to cool and solidify at room temperature. Digest logarithmically grown BT-549 and MDA-MB-231 cells with trypsin to adjust the cell concentration to 1 × 10⁻⁶ cells / well. 4 / mL, take 200μL of cell suspension and add it to a 96-well plate pre-embedded with 2% low-soluble agarose. Incubate at 37℃ in an incubator containing 5% CO2. The next day, gently aspirate 100μL of the old culture medium along the well wall with a 100μL pipette, and then add 100μL of fresh 1640 medium containing FBS. After culturing for 3 days, observe under a microscope that the tumor cells aggregate into regularly edged spherical shapes. After the 3D tumor spherical model is successfully established, discard 100μL of the old culture medium, add compound 42, and observe and record the morphology of the tumor sphericals at 0h, 24h, and 48h under a microscope.

[0154] 3D tumor spheroids can more accurately simulate the growth process of tumors in vivo. Therefore, we tested the effect of compound 42 on the formation of 3D tumor spheroids in BT-549 and MDA-MB-231 cells. The experimental results showed that compound 42 inhibited the formation of 3D tumor spheroids in a dose-dependent manner. Figure 2 Therefore, it was further confirmed that compound 42 can significantly inhibit the proliferation of triple-negative breast cancer.

[0155] BT-549 and MDA-MB-231 cells in logarithmic growth phase were seeded in 96-well plates and cultured until adherent. 2 μM of compound 42 was added to the cells for EdU medium, which was added to the wells and incubated at 37°C for 2 hours. The medium was discarded, and the cells were washed with PBS on a shaker. Cell fixation solution was added and incubated at room temperature for 30 min. The fixation solution was discarded, glycine was added, and the cells were incubated on a shaker for 5 min to decolorize. The glycine solution was discarded, and the cells were washed with PBS on a shaker. The PBS was discarded, and the cells were incubated on a shaker for 10 min with permeabilizer. The cells were then washed with PBS. Apollo staining solution was added, and the cells were incubated in the dark at room temperature on a shaker for 30 min to decolorize. The staining solution was discarded, and the cells were washed with permeabilizer on a shaker to decolorize. The permeabilizer was discarded. DAPI reaction solution was prepared, and the cells were incubated in the dark at room temperature on a shaker for 30 min to decolorize. The staining solution was discarded, and the cells were washed with PBS. The cells were observed under a fluorescence microscope, and three fields of view were randomly selected from each well for photographs.

[0156] Then, we used the EdU assay to examine the effect of compound 42 on DNA replication activity in BT-549 and MDA-MB-231 cells. Figure 5As shown, compound 42 significantly reduced the green fluorescence intensity of EdU in BT-549 and MDA-MB-231 cells, indicating that compound 42 significantly inhibited the proliferation activity of TNBC cells. Figure 3 In summary, the experimental data demonstrate that compound 42 can effectively inhibit the proliferation of TNBC cells.

[0157] Experimental Example 3: Compound 42 inhibited the migration of BT-549 and MDA-MB-231 cells.

[0158] BT-549 and MDA-MB-231 cells were digested with trypsin, placed in DMEM medium, centrifuged, and the supernatant was discarded. Culture medium was added and the cells were gently pipetted to disperse them evenly. The cells were then seeded in 6-well plates, divided into a blank control group and experimental groups (0 μM, 1 μM, 2 μM, 4 μM 42). The plates were incubated in a 5% CO2 incubator at 37°C. Cell status was observed. Once cells entered the logarithmic growth phase, cross-shaped lines were drawn in the wells using a pipette, the culture medium was aspirated, and the cells were washed three times with PBS. The cells were observed and photographed under a microscope, and the width of the scratches was recorded at 0 h. Serum-free culture medium was added, and the healing of the scratches was observed and photographed at 24 h and 48 h.

[0159] TNBC, the most common malignant tumor in women, is characterized by rapid cancer cell proliferation and a tendency to metastasize and recur. Therefore, this study investigated whether compound 42 could inhibit the migration of BT-549 and MDA-MB-231 cells. BT-549 and MDA-MB-231 cells were treated with different concentrations (0 μM, 1 μM, 2 μM, 4 μM) of compound 42 for 24 and 48 hours. Cell scratch assays were then used to verify whether compound 42 inhibited tumor cell migration. The results of the cell scratch assay showed that compound 42 inhibited the migration of BT-549 and MDA-MB-231 cells in a dose-dependent manner. Figure 4 , 5 ).

[0160] Compound 42 in Experimental Example 4 inhibited glycolysis in BT-549 and MDA-MB-231 cells by activating SIRT3.

[0161] Cellular glucose uptake was determined by flow cytometry analysis of 2-NBDG uptake. BT-549 and MDA-MB-231 cells were seeded in 96-well plates with a black transparent bottom and cultured for 24 h. After discarding the original culture medium, growth medium containing 1% FBS was added for 10 h of starvation treatment. The starvation treatment medium was then discarded, and culture medium containing 0 μM, 1 μM, 2 μM, and 4 μM of compound 42 was added. Ten minutes before the end of the intervention, the intervention medium was discarded, and 2-NBDG was added to glucose-free medium at a rate of 100 μL per well. The plates were shaken to mix thoroughly, and then incubated for another 30 min at 37°C in a humidified atmosphere containing 5% CO2 and 95% air. At the end of the culture, the samples were analyzed using an EasyCyte Plus flow cytometer system (Millipore, Chicago, IL, USA).

[0162] To measure lactate production, BT-549 and MDA-MB-231 cells were seeded in 24-well plates for 24 hours, in triplicate, and then refreshed overnight with 1 mM glucose medium. The medium was harvested, and lactate concentration was measured using lactate test strips and a lactate titrator (Nova Biomedical). Lactic acid production was standardized by cellular protein quality.

[0163] To verify whether compound 42 inhibits glycolysis in TNBC cells by activating SIRT3, we examined the glucose uptake and lactate production of BT-549 and MDA-MB-231 cells after treatment with different concentrations (0 μM, 1 μM, 2 μM, 4 μM) of compound 42 for 24 h. Glucose uptake and lactate production are key characteristics of glycolysis. The results showed that compound 42 inhibited glucose uptake in BT-549 and MDA-MB-231 cells in a dose-dependent manner. Figure 6 A, C) and lactic acid production ( Figure 6 (B, D). In short, compound 42 can inhibit glycolysis in TNBC cells by activating SIRT3.

[0164] In Experiment 5, compound 42 induced mitochondrial metabolic abnormalities and apoptosis in BT-549 and MDA-MB-231 cells by activating SIRT3.

[0165] Prepare the JC-1 working solution according to the kit instructions. Seed BT-549 and MDA-MB-231 cells in logarithmic growth phase into 6-well plates and incubate at 37°C with 5% CO2. When cells adhere and reach 80% confluence, replace the medium with medium containing (0 μM, 1 μM, 2 μM, 4 μM) compound 42. Perform triple replicates for each group and incubate again for 48 hours. Wash cells three times with PBS, trypsinize, and terminate digestion with complete culture medium. Collect cells in 1.5 mL centrifuge tubes. Centrifuge at 1000 rpm / min for 5 min at 4°C, discard the supernatant, resuspend in 500 μL of JC-1 solution, and incubate for 15 min. Centrifuge again, discard the supernatant containing JC-1, and wash twice with 1×Incubation Buffer. Finally, add 500 μL of 1×Incubation Buffer, gently resuspend the cells, and then perform flow cytometry analysis.

[0166] Logarithmically growing BT-549 and MDA-MB-231 cells were seeded in 6-well plates and cultured in a 5% CO2 incubator at 37°C. Once the cells adhered, the medium was replaced with different concentrations of compound 42: 0 μM, 1 μM, 2 μM, and 4 μM. Each group was in triplicate, and incubated at 37°C for 48 hours. The medium was removed, and the cells were washed three times with PBS, digested with trypsin, and digested with complete medium to stop the digestion. Cells were collected in centrifuge tubes, centrifuged, and the supernatant was discarded. Extraction buffer was added according to the manufacturer's instructions, and the cells were thoroughly mixed. Cells were sonicated for 1 min, centrifuged at 10000g for 10 min at 4°C, and the supernatant was transferred to a new EP tube. 500 μL of chloroform was added, and the mixture was thoroughly mixed. The tubes were centrifuged again for 3 min, and the supernatant was collected and stored on ice. ATP standards were diluted according to the manufacturer's instructions to prepare working solutions, and ATP levels were measured using a microplate reader.

[0167] Because tumor cell mitochondrial proteins are highly acetylated, we then investigated the effects of compound 42 on mitochondria in BT-549 and MDA-MB-231 cells. First, BT-549 and MDA-MB-231 cells were treated with different concentrations (0 μM, 1 μM, 2 μM, 4 μM) of compound 42 for 24 h, and then cell membrane potential was measured by flow cytometry (JC-1). The JC-1 results showed that compound 42 induced a dose-dependent decrease in mitochondrial membrane potential in both BT-549 and MDA-MB-231 cells. Figure 7 A, C). Simultaneously, we further examined the ATP content in the cells. Experimental results showed that compound 42 significantly reduced the ATP content in BT-549 and MDA-MB-231 cells (A, C). Figure 7 (B, D)

[0168] Since apoptosis is the most classic and widely studied cell death model, we then investigated whether compound 42 could induce apoptosis in BT-549 and MDA-MB-231 cells. After treating BT-549 and MDA-MB-231 cells with different concentrations (0 μM, 1 μM, 2 μM, 4 μM) of compound 42 for 24 h, Annexin V-PI staining was performed by flow cytometry. The Annexin V-PI staining results showed that compound 42 induced apoptosis in BT-549 and MDA-MB-231 cells in a dose-dependent manner. Figure 8 ).

[0169] Study on the in vivo anti-TNBC activity of compound 42 in Experimental Example 6

[0170] BALB / c mice (female, 3-4 weeks old, 19±1g) were provided by Spifort (Beijing) Biotechnology Co., Ltd. (Certificate No.: SCXK[Jing]2019-0010). Based on in vitro cytotoxicity assays and preliminary experimental results, the following treatment regimen was determined: Under standard experimental conditions, after the nude mice had adapted to the new environment and been normally fed for one week, 12 female BALB / c mice were subcutaneously inoculated with 5×10⁶ BT-549 cells. When the tumor volume reached 0.2 cm⁻¹... 3 Mice were randomly divided into two groups: a control group (n=6) and a treatment group (n=6, 20 mg / kg intraperitoneally, daily). Throughout the experiment, tumor volume and body weight were recorded daily. After 7 days of treatment, mice were euthanized by cervical dislocation, tumor tissue was collected, and its volume and weight were recorded. The tissue was then fixed and preserved in 4% paraformaldehyde. Heart, liver, spleen, lungs, and kidneys were also harvested and their weights recorded.

[0171] The antitumor activity of compound 42 in nude mice xenografted with BT-549 cells was evaluated based on tumor growth rate, tumor volume, and tumor weight. The experimental results showed that the tumor proliferation rate and volume indicated that compound 42 could effectively slow down the proliferation of BT-549 tumor cells. Figure 9 Next, the effect of compound 42 on the proliferation marker Ki-67 in tumor tissue was evaluated by immunohistochemistry. The results showed that the expression of the proliferation marker Ki-67 was reduced after treatment with compound 42, and the difference was statistically significant. Figure 9 EF). Daily body weight and organ ratios in animals showed that compound 42 had no significant toxic effects on nude mice xenografted with BT-549 cells. Figure 10 ).

Claims

1. Compounds with the following structures or pharmaceutically acceptable salts thereof: 。 2. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

3. The use of the compound of claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 2 in the preparation of a SIRT3 agonist.

4. The use of the compound of claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 2 in the preparation of an antitumor drug, wherein the tumor is triple-negative breast cancer.

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