Compound capable of degrading MYC in targeted manner and application thereof

Abstract

The invention discloses a compound capable of degrading MYC in a targeted manner and application of the compound, and belongs to the technical field of medicines. The compound can degrade MYC in a targeted manner after entering the body of a patient, so that proliferation of tumor cells such as lymphoma and osteosarcoma and cancer cells such as colorectal cancer is effectively inhibited, and the compound has a good anti-tumor effect and an anti-cancer effect; the compound can also inhibit the proliferation of myeloid monocyte leukemia cells in the body of a patient, and a new treatment scheme is provided for treating myeloid monocyte leukemia; when the compound is combined with doxorubicin, a synergistic effect is achieved, and the compound has better anti-tumor activity and has a wide application prospect in the field of medicine and pharmacology.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and particularly relates to a compound capable of targeting and degrading MYC and its application. Background Art

[0002] The occurrence of many diseases is caused by the mutation or accumulation of specific proteins. Specifically degrading pathogenic proteins can fundamentally intervene in or treat diseases. Targeted protein degradation (TPD) hijacks the endogenous protein degradation mechanism to induce the consumption or reduction of pathogenic proteins, thereby providing a new and highly potential intervention method different from traditional drugs. This new type of drug development includes proteolysis-targeting chimera (PROTAC), molecular glue, lysosome-targeting chimeras (LYTAC), autophagosome-binding compounds (Perspectives of autophagy-tethering compounds, ATTEC), autophagy-targeting chimeras (Autophagy-targeting chimeras, AUTAC and AUTOTAC), degrader-antibody conjugates (DAC), and other types. Targeted protein degradation currently mainly degrades target proteins through the ubiquitin-proteasome and lysosome. Among them, the fastest developing are molecular glue and PROTAC technologies. PROTAC uses a linker to chemically connect a target protein ligand and an E3 ligase ligand, "recruits" the E3 ligase to the vicinity of the target protein, and uses the intracellular ubiquitin-proteasome system to achieve ubiquitination labeling and protein degradation of the target protein. Molecular glue is a small molecule that induces proximity and can precisely control various biological processes in terms of time, such as signal transduction, transcription, chromatin regulation, and protein folding, localization, and degradation. Molecular glue has a very small molecular weight and its physical and chemical properties are convenient for optimization. Molecular glue mainly induces or stabilizes the protein interaction between the ubiquitin ligase and the substrate protein, thereby leading to protein degradation, and can degrade inaccessible target proteins without the need for a binding pocket on the target protein. In summary, the special mechanism of TPD drugs provides very good drug development prospects for these undruggable proteins.

[0003] MYC belongs to nuclear transcription factors, usually referring to c-Myc, which regulates cell growth and development, promotes cell metabolism and proliferation in normal cells. However, when the expression level and function of MYC are abnormal, excessive cell proliferation causes imbalance of the body's homeostasis. MYC can not only lead to the carcinogenesis of normal cells, but also promote the escape of cancer cells from immune responses, and is closely related to tumor recurrence, metastasis and drug resistance. In addition, MYC is also a key stem cell regulator and has the ability to "reprogram" adult cells into pluripotent stem cells. Studies have shown that the dysfunction or overexpression of MYC protein occurs in 70-80% of tumors, and this persistent high expression will abnormally activate tumor cells and become a driving factor for tumor development. Therefore, the development of MYC-targeted drugs can provide more treatment options for clinical cancer patients and has important biological and clinical significance. However, MYC has its inherent limitations as a drug target: 1) Antibody drugs cannot target it; 2) MYC has a smooth binding surface and lacks traditional drug-binding pockets, making it difficult for small molecule inhibitors to bind, and it is difficult for existing traditional drugs to directly target MYC; 3) Biotech drugs such as siRNA drugs have also failed in phase I / II clinical trials. A number of studies have shown that post-translational modification regulation of MYC, such as phosphorylation, ubiquitination and acetylation, regulates its transcriptional activity by affecting the formation of MYC transcription complexes or proteasomal degradation. Dysregulation of post-translational modification leading to excessive accumulation of MYC is the key reason for the activation of downstream proliferation, metastasis and stemness signaling pathways, suggesting that targeting MYC degradation is a new strategy for inhibiting tumors. Therefore, it is necessary to develop a compound that can target and degrade MYC to solve the above problems. Summary of the Invention

[0004] Aiming at the deficiencies in the prior art, the purpose of the present invention is to provide a compound that can target and degrade MYC, so as to provide a new treatment plan for cancer, tumors and leukemia.

[0005] To achieve the above purpose, the present invention adopts the following technical solutions: A compound that can target and degrade MYC, the structure of the compound is: ; wherein, R 1Selected from acetyl, o-fluorobenzoyl, m-fluorobenzoyl, p-fluorobenzoyl, 2,6-difluorobenzoyl, cyclohexanecarbonyl, cyclopentanecarbonyl, 2-(piperidin-1-yl)acetyl, 2-(4-methylpiperazin-1-yl)acetyl, cyclopropanecarbonyl, cyclobutanecarbonyl, 2,2,3,3-tetramethylcyclopropane-1-carbonyl, 2-morpholinoacetyl, 3-methylbutyryl, pyrimidin-2-yl, 3,3-dimethylbutyryl, 3,5-difluorobenzoyl, hydrogen, p-carboxybenzoyl, 3-hydroxy-3-methylbutyryl, thiazol-2-ylcarbonyl, 3-methyl-2-((methylformamido)amino)butyryl, 2-amino-pyrimidin-5-ylcarbonyl, 3-amino-3-methylbutyryl, 3,4-dimethoxybenzoyl, 3,4,5-trimethoxybenzoyl, 4-amino-5-(ethylsulfonyl)-benzoyl, 2-(4-(4-methylpiperazin-1-yl)piperidine)acetyl, 4-hydroxy-2,6-dimethylbenzoyl, 4-bromothiazol-2-ylcarbonyl, 4-cyclopropylthiazol-2-ylcarbonyl, 4,5-dimethylthiazol-2-ylcarbonyl, 5-methylthiazol-2-ylcarbonyl, 2-aminopyrimidin-5-ylcarbonyl, 4-hydroxy-2-methylbenzoyl, 4-amino-2-methylbenzoyl, 4-amino-2-methoxybenzoyl, 3-methyl-1H-pyrazol-4-ylcarbonyl, 3,5-dimethyl-1H-pyrazol-4-ylcarbonyl, 3-methyl-2-((tert-butoxycarbonyl)amino)butyramide, 4-amino-4-methyl-valeryl, 4-amino-3,5-dimethylbenzoyl, 2-aminoacetyl, 4-amino-5-chloro-2-methoxybenzoyl, 4-amino-5-chloro-2,3-dihydrobenzofuran-7-ylcarbonyl, 7-methoxy-4-oxo-1,4-dihydroquinoline-6-ylcarbonyl, 4-amino-5-chloro-2-ethoxybenzoyl, 5-chloro-2-methoxy-4-(methylamino)benzoyl, 4-amino-2,3,5,6-tetrafluorobenzoyl, -2,5-dimethoxy-3-nitrobenzoyl, oxazol-4-ylcarbonyl, 3,5-diamino-6-chloropyrazine-2-ylcarbonyl, oxazol-4-carboxamide, 1H-imidazol-2-ylcarbonyl, pyrimidin-2-yl, thiazol-2-yl, 1H-imidazol-2-yl, 2,4(1H,3H)-dioxopyrimidin-6-yl, 1-methyl-1H-1,2,3-triazol-4-yl, 5-methylthiazol-4-yl, 3-methylthiazol-2-yl, 5-chlorothiazol-2-yl, 4,5-dimethylthiazol-2-yl, 4-chlorothiazol-2-yl, 2-methylthiazol-4-yl, 1,3-dimethyl-1H-pyrazol-4-yl, 5-phenyloxazol-2-yl, 4,5-dimethyloxazol-2-yl, 5-methylformylthiazol-4-yl, 5-thiazol-4-carboxylic acid group, oxazol-2-yl, 4-methylthiazol-2-yl, 5-methyl-1,3,4-oxadiazol-2-yl, 5-phenyl-1,3,4 - Oxadiazol - 2 - yl, oxazole - 5 - carboxylic acid ethyl ester - 2 - yl, 2 - (trifluoromethyl)pyrimidine - 5 - carbonyl, 4 - amino - 2 - methylpyrimidine - 5 - carbonyl, 5 - amino - pyridazine - 4 - carbonyl, 2 - amino - 4 - (trifluoromethyl)pyrimidine - 5 - carbonyl, 6 - amino - pyrazine - 2 - carbonyl, 3 - amino - 2 - methoxy - isonicotinoyl, oxadiazol - 4 - yl, 2,2,3,3 - tetramethylcyclopropane - 1 - carbonyl, 3,3 - dimethylbutyryl, 3,3 - dimethylbutyryl, 2 - cyclopropyl - 4 - fluorobenzene - 1 - carbonyl, 2 - fluorobenzene - 1 - carbonyl, 3,3 - dimethyl - butyryl, 2 - (methoxycarbonyl)aminobutyryl, 2,4 - dimethylbenzoyl, 2 - ((3 - aminopyrazin - 2 - yl)thio)acetyl, 4 - ((tert - butoxycarbonyl)amino)benzoyl, 4 - amino - 5 - (ethylsulfonyl)-2 - methoxybenzoyl, methyl formate - 1 - yl, ethyl formate - 1 - yl, 2,5 - dichloro - thiazole - 4 - carbonyl, 5 - fluoro - 2 - hydroxy - nicotinoyl, 6 - hydroxy - 2 - methyl - nicotinoyl, 5 - oxo - 4,5 - dihydro - 1H - 1,2,4 - triazole - 3 - carbonyl, 2 - hydroxy - 6 - methylisonicotinoyl, 1,2,5 - oxadiazole - 3 - nitrile - 6 - yl; R 2 selected from 2 - cyclopropyl - 4 - fluorophenyl, 2 - cyclopentyl - 4 - fluorophenyl, 5 - fluoro - 4'-methyl - [1,1'-biphenyl]-2 - yl, 2 - cyclohexyl - 4 - fluorophenyl, 2 - hydroxy - 4 - fluorophenyl, 2 - methyl - 4 - fluorophenyl, 2 - methoxy - 4 - carboxyphenyl, 2 - cyclopropyl - 4 - carbonylphenyl, 2 - methyl - 5 - carboxyphenyl, neopentyl, 2 - cyclohexyl - 4 - formylphenyl, 2 - cyclohexyl - 5 - carboxyphenyl, 2 - methyl - 4 - fluoro - 5 - carboxyphenyl, 5 - fluoro - 4'-carboxy - [1,1'-biphenyl]-2 - yl.

[0006] The present invention also provides a compound capable of targeting and degrading MYC and its applications, and the drug includes the compound as described above.

[0007] Furthermore, the drug also includes pharmaceutically acceptable drug excipients, and the drug excipients are pharmaceutically acceptable salts, excipients or carriers.

[0008] Furthermore, the carrier is any one or more of fillers, wetting agents, binders, disintegrants or lubricants.

[0009] Furthermore, the dosage form of the pharmaceutical composition is any one or more of water infusions, powders, lotions, tinctures, oils, emulsions, ointments, plasters or aerosols.

[0010] Another object of the present invention is to provide the use of the compound capable of targetedly degrading MYC as described above in the preparation of anti-tumor drugs, anti-cancer drugs, drugs for treating leukemia, and drugs for anti-tumor in combination with doxorubicin.

[0011] The beneficial effects of the present invention are as follows: The present invention provides a compound capable of targetedly degrading MYC. After entering the patient's body, the compound can targetedly degrade MYC, thereby effectively inhibiting the proliferation of tumor cells such as lymphoma and osteosarcoma and cancer cells such as colorectal cancer, and having good anti-tumor and anti-cancer effects; the compound can also inhibit the proliferation of myelomonocytic leukemia cells in the patient's body, providing a new treatment plan for treating myelomonocytic leukemia; the compound has a synergistic effect when combined with doxorubicin and has better anti-tumor activity, and has broad application prospects in the field of medicine.

[0012] The above description is only an overview of the technical solution of the present invention. In order to be able to understand the technical means of the present invention more clearly and implement it in accordance with the content of the specification, the following will describe in detail with reference to the preferred embodiments of the present invention and the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings required for the description of the specific embodiments or the prior art. Obviously, some of the following drawings are embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative efforts.

[0014] Figure 1 It is a time- and dose-dependent statistical chart for detecting the degradation of MYC protein expression by a series of compounds in Daudi cells at different times and concentrations using Western blot technology; Figure 2 It is a result chart for detecting the inhibitory effect of compound 8 at different concentrations on the tumor growth of Daudi cells in an in vivo study by constructing a Daudi subcutaneous tumor model; Figure 3 It is for constructing a Daudi advanced-stage transplanted tumor model (>500 mm 3 ) to detect the result chart of the inhibitory effect of compound 30 on the tumor growth of Daudi cells; Figure 4 It is a result chart for detecting the anti-tumor effect of the combined treatment of compound 30 and the lymphoma first-line drug doxorubicin by constructing a Daudi cell subcutaneous tumor model. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0015] To better describe the present invention, the following provides further illustration through specific embodiments. Unless otherwise specified, the methods in the following embodiments are all conventional methods.

[0016] Unless otherwise specified, the technical solutions of the present invention are all conventional solutions in the art; unless otherwise specified, the reagents or materials are all from commercial channels.

[0017] The following embodiments are directed to the intermediate compounds and final products determined in the specification and synthetic schemes. The following embodiments are used to describe in detail the preparation of the compounds of the present invention, but the described chemical reactions are disclosed according to their general applicability to the preparation of the compounds of the present invention. Sometimes, the described reaction may not be applicable to each compound within the scope of the present invention as described. Those skilled in the art can easily identify the compounds for which this occurs. In these cases, the reaction can be successfully carried out by conventional improvements known to those skilled in the art. In all preparation methods, all raw materials are known or can be easily prepared using known raw materials. All temperatures are given in degrees Celsius, unless otherwise explicitly stated. When referring to yields, all parts and percentages are based on the amount of substance. When referring to solvents and eluents, all parts are by volume.

[0018] The abbreviations used in the following embodiments are as follows: Et 3 N is triethylamine; Toluene is toluene; Pd(PPh 3 ) 4 is tetrakis(triphenylphosphine)palladium; Na 2 CO 3 is sodium carbonate; ACN is acetonitrile; HATU is 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; DMF is dimethylformamide; PBS is phosphate buffered saline; K 2 CO 3 is potassium carbonate; Boc is tert-butoxycarbonyl; OH is hydroxyl; TFA is trifluoroacetic acid; DCM is dichloromethane; Pyridine is pyridine; AcOH is acetic acid; PBST is phosphate buffered saline; DMSO is dimethyl sulfoxide; PVDF is polyvinylidene fluoride; GAPDH is glyceraldehyde-3-phosphate dehydrogenase; PEG is polyethylene glycol.

[0019] Example 1 Synthesis of Compound 1 ; N-(2-Cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)acetamide Synthesis Scheme: ; Step 1: Synthesis of 2-Cyclopropyl-4-fluoroaniline ; Add cyclopropylboronic acid (4.97 g, 57.89 mmol), 2-bromo-4-fluoroaniline (10.00 g, 52.63 mmol), potassium phosphate (27.93 g, 131.57 mmol), toluene (100 ml), and purified water (20 ml) into the reaction flask. Stir at room temperature for 10 min. Then add palladium acetate (0.60 g, 2.63 mmol) and tricyclohexylphosphine (1.48 g, 5.26 mmol) into the reaction flask. Replace nitrogen, and stir the reaction at 90 °C for 8 hours. After the reaction is completed, cool to room temperature, evaporate the solvent, extract with dichloromethane, concentrate, and purify by column chromatography to obtain 2-cyclopropyl-4-fluoroaniline as a brown liquid (7.00 g, yield 88%, purity 95.3%). Characterization: LCMS m / z = 152.08 [M+1] + 。

[0020] Step 2: Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine ; Add methyl 6-amino-4'-methyl-[1,1'-biphenyl]-3-carboxylate (7.00 g, 46.30 mmol), 4-fluoro-7-nitrobenzo[c][1,2,5]oxadiazole (9.33 g, 50.93 mmol), acetonitrile (28 ml) and purified water (7 ml) into the reaction flask. Stir the reaction at 75 °C for 8 h. After the reaction is complete, cool to room temperature, and a large amount of red solid precipitates. Filter and wash with acetonitrile to obtain N-(2-cyclopropyl-4-fluorophenyl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine as a bright red solid (14.26 g, yield: 98%, purity 99.1%). Characterization: LCMS m / z = 315.08 [M+1]+.

[0021] Step 3: Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)acetamide ; Add acetic acid (76 mg, 1.27 mmol), TCTU (566 mg, 1.59 mmol), DMF (2 ml), and triethylamine (193 mg, 1.91 mmol) to the reaction flask respectively, and stir at room temperature for 5 min. Add N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)acetamide (200 mg, 0.64 mmol) to the reaction solution, stir at room temperature for 4 h, dilute with purified water, extract with dichloromethane, concentrate the organic phase to obtain a brown liquid, and purify by column chromatography (n-heptane:ethyl acetate = 1:2) to obtain an oily substance. Purify by preparative HPLC to obtain N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)acetamide, which is a light yellow solid (122 mg, yield: 54%, purity 100%). Characterization: LCMS m / z = 357.09 [M+1] + 。

[0022] Synthesis of Compound 2 in Example 2 ; Synthesis of N-(2-cyclopropyl-4-fluorophenyl)-2-fluoro-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide Prepare N-(2-cyclopropyl-4-fluorophenyl)-2-fluoro-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide in a similar manner to Steps 1-3 of Example 1. Characterization: LCMS m / z = 437.10 [M+1] +

[0023] Synthesis of Compound 3 in Example 3 ; Synthesis of N-(2-cyclopropyl-4-fluorophenyl)-3-fluoro-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide Prepare N-(2-cyclopropyl-4-fluorophenyl)-3-fluoro-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide in a similar manner to Steps 1-3 of Example 1. Characterization: LCMS m / z = 437.10 [M+1] +

[0024] Synthesis of Compound 4 in Example 4 ; Synthesis of N-(2-cyclopropyl-4-fluorophenyl)-2,6-difluoro-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide Prepare N-(2-cyclopropyl-4-fluorophenyl)-2,6-difluoro-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide in a similar manner to Steps 1-3 of Example 1. Characterization: LCMS m / z = 455.09 [M+1] +

[0025] Example 5 Synthesis of Compound 5 ; Synthesis of N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclohexanecarboxamide Prepare N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclohexanecarboxamide in a similar manner to Steps 1-3 of Example 1. Characterization: 1 H NMR (400 MHz, CDCl3) δ 8.41 (d, J = 8.0 Hz, 1H), 7.11 (d, J = 8.0Hz, 1H), 6.98 (ddd, J = 8.6, 7.6, 2.9 Hz, 1H), 6.66 (dd, J = 9.7, 2.9 Hz,1H), 2.48 – 2.28 (m, 1H), 1.97 – 1.53 (m, 12H), 1.37 – 1.19 (m, 4H), 1.17 –0.95 (m, 3H), 0.81 – 0.69 (m, 2H), 0.67 – 0.52 (m, 1H). LCMS m / z = 425.15[M+1] +

[0026] Example 6 Synthesis of Compound 6 Synthesis of N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopentanecarboxamide.

[0027] Prepare N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopentanecarboxamide in a similar manner to Steps 1-3 of Example 1. Characterization: LCMS m / z = 411.14 [M+1] + ; 。

[0028] Example 7 Synthesis of Compound 7 ; Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)-2-(piperidin-1-yl)acetamide Prepare N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)-2-(piperidin-1-yl)acetamide in a similar manner to Steps 1-3 of Example 1. Characterization: LCMS m / z = 440.17 [M+1] +

[0029] Example 8 Synthesis of Compound 8 ; Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-2-(4-methylpiperazin-1-yl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)acetamide Prepare N-(2-cyclopropyl-4-fluorophenyl)-2-(4-methylpiperazin-1-yl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)acetamide in a similar manner to Steps 1-3 of Example 1. Characterization: LCMS m / z = 455.18 [M+1] +

[0030] Example 9 Synthesis of Compound 9 ; Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopropanecarboxamide Prepare N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopropanecarboxamide in a similar manner to Steps 1-3 of Example 1. Characterization: LCMS m / z = 383.11[M+1] +

[0031] Example 10 Synthesis of Compound 10 ; Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclobutanecarboxamide Prepare N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclobutanecarboxamide in a similar manner to Steps 1-3 of Example 1. Characterization: LCMS m / z = 397.12 [M+1] +

[0032] Example 11 Synthesis of Compound 11 Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-2,2,3,3-tetramethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopropane-1-carboxamide ; Prepare N-(2-Cyclopropyl-4-fluorophenyl)-2,2,3,3-tetramethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopropane-1-carboxamide in a similar manner to Steps 1-3 of Example 1. Characterization: 1 H NMR (400 MHz, CDCl3) δ 8.42 (d, J = 8.1 Hz, 1H), 7.10 (d, J = 8.1Hz, 1H), 7.05 – 6.97 (td, 1H), 6.67 (dd, J = 9.7, 2.9 Hz, 1H), 1.88 – 1.76(m, 1H), 1.39 (s, 3H), 1.32 (s, 4H), 1.08 (s, 3H), 1.01 (s, 5H), 0.79 (ddd, J= 11.3, 9.9, 5.1 Hz, 1H), 0.71 – 0.61 (m, 1H), 0.43 (ddt, J = 9.6, 6.5, 4.9Hz, 1H).LCMS m / z=438.17 [M+1] +

[0033] Example 12 Synthesis of Compound 12 ; Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-2-morpholinyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)acetamide Prepare N-(2-Cyclopropyl-4-fluorophenyl)-2-morpholinyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)acetamide in a similar manner to Steps 1-3 of Example 1. Characterization: 11H NMR (400 MHz, CDCl3) δ 8.48 (d, J = 8.0 Hz, 1H), 7.49 (dd, J =8.4, 5.5 Hz, 2H), 7.02 (ddd, J = 8.7, 7.5, 2.8 Hz, 1H), 6.66 (dd, J = 9.5,2.8 Hz, 1H), 4.22 (d, J = 16.4 Hz, 1H), 4.05 (t, J = 4.7 Hz, 4H), 3.78 (d, J= 16.4 Hz, 1H), 1.97 – 1.87 (m, 1H), 1.18 – 1.07 (m, 1H), 0.86 – 0.72 (m,2H), 0.62 – 0.49 (m, 1H). LCMS m / z=442.14 [M+1] +

[0034] Example 13 Synthesis of Compound 13 ; Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-3-methyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)butanamide N-(2-Cyclopropyl-4-fluorophenyl)-3-methyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)butanamide was prepared in a similar manner to Steps 1-3 of Example 1. Characterization results are as follows: 1 1H NMR (400 MHz, CDCl3) δ 8.44 (d, J = 8.0 Hz, 1H), 7.28 – 7.21 (m,1H), 7.14 (d, J = 8.0 Hz, 1H), 7.00 (ddd, J = 8.6, 7.6, 2.9 Hz, 1H), 6.66(dd, J = 9.7, 2.9 Hz, 1H), 2.29 (q, J = 14.8 Hz, 2H), 1.95 – 1.85 (m, 1H),1.15 (s, 8H), 1.12 – 1.05 (m, 1H), 0.84 – 0.74 (m, 2H), 0.65 – 0.56 (m, 1H). LCMS m / z=399.14 [M+1] +

[0035] Example 14 Synthesis of Compound 14 ; Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-7-nitro-N-(pyrimidin-2-yl)benzo[c][1,2,5]oxadiazol-4-amine N-(2-Cyclopropyl-4-fluorophenyl)-7-nitro-N-(pyrimidin-2-yl)benzo[c][1,2,5]oxadiazol-4-amine was prepared in a similar manner to Steps 1-3 of Example 1. Characterization: LCMS m / z = 393.10 [M+1] +

[0036] Example 15 Synthesis of Compound 15 Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-3,3-dimethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)butyramide ; N-(2-Cyclopropyl-4-fluorophenyl)-3,3-dimethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)butyramide was prepared in a similar manner to Steps 1-3 of Example 1. Characterization: LCMS m / z = 413.15 [M+1] +

[0037] Example 16 Synthesis of Compound 16 ; Synthesis of N-(2-Cyclohexyl-4-fluorophenyl)-3,3-dimethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)butyramide N-(2-Cyclohexyl-4-fluorophenyl)-3,3-dimethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)butyramide was prepared in a similar manner to Steps 1-3 of Example 1. Characterization: 11H NMR (400 MHz, CDCl3) δ 8.42 (d, J = 8.0 Hz, 1H), 7.35 (dd, J = 8.6, 5.3 Hz, 1H), 7.15 – 7.05 (m, 2H), 7.04 (d, J = 8.0 Hz, 1H), 2.74 (td, J = 11.9, 2.0 Hz, 1H), 2.28 – 2.15 (m, 2H), 1.46 (ddd, J = 12.5, 10.3, 6.4 Hz, 2H), 1.28 (s, 3H), 1.14 (s, 11H), 0.89 (dd, J = 20.1, 9.5 Hz, 2H), 0.70 – 0.55 (m, 1H). LCMS m / z = 455.20 [M+1] +

[0038] Example 17 Synthesis of Compound 17 ; Synthesis of N-(2-Cyclohexyl-4-fluorophenyl)-3,5-difluoro-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide Prepare N-(2-Cyclohexyl-4-fluorophenyl)-3,3-dimethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)butanamide in a similar manner to Steps 1-4 of Example 1. Characterization: LCMS m / z = 455.09 [M+1] +

[0039] Example 18 Synthesis of Compound 18 ; Synthesis of 5-Fluoro-2-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)phenol Prepare 5-Fluoro-2-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)phenol in a similar manner to Steps 1-4 of Example 1. Characterization: LCMS m / z = 291.05 [M+1] +

[0040] Example 19 Synthesis of Compound 19 ; Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-4-fluoro-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide Prepare N-(2-cyclopropyl-4-fluorophenyl)-4-fluoro-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide in a similar manner to Steps 1 and 3-4 of Example 1. Characterization: LCMS m / z = 437.10 [M+1] +

[0041] Example 20 Synthesis of Compound 20 ; Synthesis of 4-((2-cyclopropyl-4-fluorophenyl)(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)carbamoyl)benzoic acid Prepare 4-((2-cyclopropyl-4-fluorophenyl)(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)carbamoyl)benzoic acid in a similar manner to Steps 1 and 3-4 of Example 1. Characterization: LCMS m / z = 463.10 [M+1] +

[0042] Example 21 Synthesis of Compound 21 ; Synthesis of N-(2-cyclopropyl-4-fluorophenyl)-3-hydroxy-3-methyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)butyramide Prepare N-(2-cyclopropyl-4-fluorophenyl)-3-hydroxy-3-methyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)butyramide in a similar manner to Steps 1-3 of Example 1. Characterization: LCMS m / z = 415.13 [M+1] +

[0043] Example 22 Synthesis of Compound 22 ; Synthesis of N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)thiazole-2-carboxamide Prepare N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)thiazole-2-carboxamide in a similar manner to Steps 1-3 of Example 1. Characterization: 11H NMR (600 MHz, DMSO) δ 9.81 (s, 1H), 9.08 (s, 1H), 8.49 (d, J = 8.9Hz, 1H), 7.33 (dd, J = 8.8, 5.7 Hz, 1H), 6.96 (td, J = 8.5, 2.9 Hz, 1H), 6.76– 6.68 (m, 2H), 2.90 (s, 2H), 1.93 (td, J = 8.4, 4.4 Hz, 1H), 1.65 (s, 6H),0.86 (tt, J = 12.2, 6.1 Hz, 2H), 0.67 – 0.62 (m, 2H).LCMS m / z=426.06 [M+1] +

[0044] Example 23 Synthesis of Compound 23 ; Synthesis of Methyl (1-(((2-Cyclopropyl-4-fluorophenyl)(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate Methyl (1-(((2-Cyclopropyl-4-fluorophenyl)(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate was prepared in a similar manner to Steps 1-3 of Example 1. Characterization: LCMS m / z=472.16 [M+1] +

[0045] Example 24 Synthesis of Compound 24 ; Synthesis of 2-Amino-N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)pyrimidine-5-carboxamide 2-Amino-N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)pyrimidine-5-carboxamide was prepared in a similar manner to Steps 1-3 of Example 1. Characterization: LCMS m / z=436.11 [M+1] +

[0046] Example 25 Synthesis of Compound 25

[0047] Synthesis of 3-amino-N-(2-cyclopropyl-4-fluorophenyl)-3-methyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)butanamide 3-Amino-N-(2-cyclopropyl-4-fluorophenyl)-3-methyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)butanamide was prepared in a similar manner to Steps 1-3 of Example 1. Characterization was as follows: 1 H NMR (400 MHz, CDCl3) δ 8.46 (d, J = 8.0 Hz, 1H), 7.57 (q, J = 3.1Hz, 2H), 7.10 (d, J = 8.0 Hz, 1H), 6.94 (ddd, J = 8.6, 7.7, 2.9 Hz, 1H), 6.74(dd, J = 9.7, 2.9 Hz, 1H), 1.96 – 1.88 (m, 1H), 0.89 – 0.82 (m, 2H), 0.77 –0.69 (m, 1H), 0.64 – 0.56 (m, 1H), 0.47 (ddt, J = 9.7, 6.4, 4.9 Hz, 1H).LCMSm / z=414.16 [M+1] +

[0048] Example 26 Synthesis of Compound 26 ; Synthesis of N-(2-cyclopropyl-4-fluorophenyl)-3,4-dimethoxy-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide N-(2-Cyclopropyl-4-fluorophenyl)-3,4-dimethoxy-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide was prepared in a similar manner to Step 1 and Steps 3-4 of Example 1. Characterization was as follows: 11H NMR (400 MHz, CDCl3) δ 8.41 (d, J = 8.0 Hz, 1H), 7.23 (m, 1H), 7.12 (dd, J = 8.7, 5.2 Hz, 1H), 6.90 – 6.84 (m, 2H), 6.73 (d, J = 8.4 Hz, 1H), 6.62 (dd, J = 9.7, 2.8 Hz, 1H), 3.88 (s, 3H), 3.79 (s, 3H), 1.87 (ddd, J = 14.2, 8.9, 5.6 Hz, 2H), 0.98 – 0.91 (m, 1H), 0.73 (m, 1H), 0.63 – 0.55 (m, 1H), 0.47 – 0.39 (m, 1H). LCMS m / z = 479.13 [M+1] +

[0049] Example 27 Synthesis of Compound 27 ; Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-3,4,5-trimethoxy-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide Prepare N-(2-cyclopropyl-4-fluorophenyl)-3,4,5-trimethoxy-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide in a similar manner to Steps 1-3 of Example 1. Characterization results are as follows: 1 1H NMR (400 MHz, CDCl3) δ 8.43 (d, J = 8.0 Hz, 1H), 7.12 (dd, J = 8.7, 5.2 Hz, 1H), 6.94 – 6.85 (m, 4H), 6.60 (dd, J = 9.6, 2.8 Hz, 1H), 3.86 (s, 3H), 3.72 (s, 6H), 1.94 – 1.85 (m, 1H), 0.99 – 0.91 (m, 1H), 0.73 (tt, J = 10.0, 5.2 Hz, 1H), 0.64 – 0.56 (m, 1H), 0.43 (ddd, J = 11.3, 9.6, 4.9 Hz, 1H). LCMS m / z = 509.14 [M+1] +

[0050] Example 28 Synthesis of Compound 28 ; Synthesis of 4-Amino-N-(2-cyclopropyl-4-fluorophenyl)-5-(ethylsulfonyl)-2-methoxy-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide Prepare 4-Amino-N-(2-cyclopropyl-4-fluorophenyl)-5-(ethylsulfonyl)-2-methoxy-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide according to the steps of Step 1-3 of Example 1. Characterization: 1 H NMR (600 MHz, DMSO) δ 8.59 (d, J = 8.1 Hz, 1H), 7.73 (s, 1H), 7.27(dd, J = 8.7, 5.5 Hz, 1H), 7.20 (d, J = 8.1 Hz, 1H), 6.98 (td, J = 8.4, 2.8Hz, 1H), 6.81 (dd, J = 10.3, 2.8 Hz, 1H), 6.52 (s, 2H), 6.20 (s, 1H), 3.42(s, 3H), 3.14 (q, J = 7.3 Hz, 2H), 1.92 – 1.88 (m, 1H), 1.01 (t, J = 7.3 Hz,3H), 0.88 – 0.80 (m, 4H), 0.55 – 0.48 (m, 1H), 0.33 (s, 1H).LCMS m / z=556.12[M+1] +

[0051] Example 29 Synthesis of Compound 29 ; Synthesis of 4-Bromo-N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)thiazole-2-carboxamide Prepare 4-Bromo-N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)thiazole-2-carboxamide according to the steps of Step 1-3 of Example 1. Characterization: 11H NMR (600 MHz, CDCl3) δ 8.47 (d, J = 8.0 Hz, 1H), 7.45 (s, 1H), 7.15 (d, J = 8.0 Hz, 1H), 6.97 – 6.93 (m, 1H), 6.73 (dd, J = 9.7, 2.8 Hz, 1H), 1.92 – 1.86 (m, 1H), 0.92 – 0.86 (m, 1H), 0.74 – 0.70 (m, 1H), 0.66 – 0.61 (m, 1H), 0.54 – 0.49 (m, 1H). LCMS m / z = 505.98 [M+1] +

[0052] Example 30 Synthesis of Compound 30 ; Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-4,5-dimethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)thiazole-2-carboxamide N-(2-Cyclopropyl-4-fluorophenyl)-4,5-dimethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)thiazole-2-carboxamide was prepared according to the steps of Steps 1-3 of Example 1. Characterization: LCMS m / z = 454.09 [M+1] +

[0053] Example 31 Synthesis of Compound 31 ; Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-5-methyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)thiazole-2-carboxamide N-(2-Cyclopropyl-4-fluorophenyl)-5-methyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)thiazole-2-carboxamide was prepared according to the steps of Steps 1-3 of Example 1. Characterization: LCMS m / z = 440.09 [M+1] +

[0054] Example 32 Synthesis of Compound 32 ; Synthesis of 2-Amino-N-(2-cyclopropyl-4-fluorophenyl)-4-methyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)pyrimidine-5-carboxamide Prepare 2-amino-N-(2-cyclopropyl-4-fluorophenyl)-4-methyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)pyrimidine-5-carboxamide according to the steps of Step 1-3 of Example 1. Characterization: LCMS m / z = 450.12 [M+1] +

[0055] Example 33 Synthesis of Compound 33 ; Synthesis of N-(2-cyclopropyl-4-fluorophenyl)-4-hydroxy-2-methyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide Prepare N-(2-cyclopropyl-4-fluorophenyl)-4-hydroxy-2-methyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide according to the steps of Step 1-3 of Example 1. Characterization: LCMS m / z = 449.12 [M+1] +

[0056] Example 34 Synthesis of Compound 34 ; Synthesis of 4-amino-N-(2-cyclopropyl-4-fluorophenyl)-2-methoxy-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide Prepare 4-amino-N-(2-cyclopropyl-4-fluorophenyl)-2-methoxy-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide according to the steps of Step 1-3 of Example 1. Characterization: LCMS m / z = 464.13 [M+1] +

[0057] Example 35 Synthesis of Compound 35 ; Synthesis of N-(2-cyclopropyl-4-fluorophenyl)-3-methyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)-1H-pyrazole-4-carboxamide Prepare N-(2-cyclopropyl-4-fluorophenyl)-3-methyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)-1H-pyrazole-4-carboxamide according to the steps of Step 1-3 of Example 1. Characterization: LCMS m / z = 423.11 [M+1] +

[0058] Example 36 Synthesis of Compound 36 ; Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-3,5-dimethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)-1H-pyrazole-4-carboxamide Prepare N-(2-Cyclopropyl-4-fluorophenyl)-3,5-dimethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)-1H-pyrazole-4-carboxamide according to the steps of Steps 1-3 of Example 1. Characterization: LCMS m / z = 437.13 [M+1] +

[0059] Example 37 Synthesis of Compound 37 ; Synthesis of 4-Amino-N-(2-cyclopropyl-4-fluorophenyl)-4-methyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)pentanamide Prepare 4-Amino-N-(2-cyclopropyl-4-fluorophenyl)-4-methyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)pentanamide according to the steps of Steps 1-3 of Example 1. Characterization: 1 H NMR (400 MHz, DMSO) δ 8.67 (d, J = 8.0 Hz, 1H), 7.80 (s, 3H), 7.62 (dd, J = 8.6, 5.5 Hz, 1H), 7.46 (d, J = 8.0 Hz, 1H), 7.18 (td, J = 8.4, 2.7 Hz, 1H), 6.95 (dd, J = 10.2, 2.6 Hz, 1H), 3.94 (s, 5H), 2.42 – 2.30 (m, 2H), 1.90 (dd, J = 16.3, 7.5 Hz, 4H), 1.15 (m, 7H), 0.95 (m, 1H), 0.86 – 0.78 (m, 1H), 0.66 – 0.53 (m, 2H). LCMS m / z = 428.17 [M+1] +

[0060] Example 38 Synthesis of Compound 38 ; Synthesis of 4-Amino-N-(2-cyclopropyl-4-fluorophenyl)-3,5-dimethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide Prepare 4-amino-N-(2-cyclopropyl-4-fluorophenyl)-3,5-dimethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide according to the steps of Step 1-3 in Example 1. Characterization: LCMS m / z = 462.15 [M+1] +

[0061] Example 39 Synthesis of Compound 39 ; Synthesis of 2-amino-N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)acetamide Prepare 2-amino-N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)acetamide according to the steps of Step 1-3 in Example 1. Characterization: LCMS m / z = 372.10 [M+1] +

[0062] Example 40 Synthesis of Compound 40 ; Prepare 4-amino-5-chloro-N-(2-cyclopropyl-4-fluorophenyl)-2-methoxy-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide according to the steps of Step 1-3 in Example 1.

[0063] Characterization: 1 H NMR (600 MHz, DMSO) δ 8.58 (d, J = 8.1 Hz, 1H), 7.43 (s, 1H), 7.22(dd, J = 8.6, 5.5 Hz, 1H), 7.14 (d, J = 8.1 Hz, 1H), 6.98 (td, J = 8.4, 2.8Hz, 1H), 6.81 (dd, J = 10.3, 2.8 Hz, 1H), 6.14 (s, 1H), 3.24 (s, 3H), 1.94 –1.89 (m, 1H), 0.87 – 0.78 (m, 2H), 0.55 – 0.48 (m, 1H), 0.42 – 0.34 (m, 1H). LCMS m / z = 498.09 [M+1] +

[0064] Example 41 Synthesis of Compound 41 ; Synthesis of 5-chloro-N-(2-cyclopropyl-4-fluorophenyl)-2-methoxy-4-(methylamino)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide Prepare 5-chloro-N-(2-cyclopropyl-4-fluorophenyl)-2-methoxy-4-(methylamino)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide according to the steps of steps 1-3 of Example 1. Characterization: LCMS m / z = 512.11 [M+1] +

[0065] Example 42 Synthesis of Compound 42 ; Synthesis of 4-amino-N-(2-cyclopropyl-4-fluorophenyl)-2,3,5,6-tetrafluoro-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide Prepare 4-amino-N-(2-cyclopropyl-4-fluorophenyl)-2,3,5,6-tetrafluoro-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide according to the steps of steps 1-3 of Example 1. Characterization: 1 H NMR (600 MHz, DMSO) δ 8.63 (d, J = 8.0 Hz, 1H), 7.40 – 7.33 (m,2H), 7.07 (td, J = 8.4, 2.9 Hz, 1H), 6.85 (dd, J = 10.2, 2.8 Hz, 1H), 6.65(s, 2H), 1.90 – 1.84 (m, 1H), 0.89 – 0.79 (m, 2H), 0.67 – 0.60 (m, 1H), 0.46– 0.39 (m, 1H). LCMS m / z = 506.09 [M+1] +

[0066] Example 43 Synthesis of Compound 43 ; Synthesis of N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)oxazole-4-carboxamide Prepare N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)oxazole-4-carboxamide according to the steps of steps 1-3 of Example 1. Characterization: 11H NMR (400 MHz, DMSO) δ 8.68 – 8.61 (m, 1H), 8.29 (s, 1H), 7.42 (dd, J = 8.6, 5.5 Hz, 1H), 7.28 (d, J = 8.0 Hz, 1H), 7.09 (td, J = 8.4, 2.7 Hz, 1H), 6.95 (dd, J = 10.1, 2.5 Hz, 1H), 1.89 – 1.78 (m, 1H), 1.23 (s, 1H), 0.82 – 0.68 (m, 1H), 0.58 – 0.44 (m, 1H). LCMS m / z = 410.08[M+1] +

[0067] Example 44 Synthesis of Compound 44 ; Synthesis of 3,5-diamino-6-chloro-N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)pyrazine-2-carboxamide 3,5-diamino-6-chloro-N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)pyrazine-2-carboxamide was prepared according to the steps of Step 1-3 in Example 1. Characterization: LCMS m / z = 485.08[M+1] +

[0068] Example 45 Synthesis of Compound 45 ; Synthesis of N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)-1H-imidazole-2-carboxamide N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)-1H-imidazole-2-carboxamide was prepared according to the steps of Step 1-3 in Example 1. Characterization: LCMS m / z = 409.10[M+1] +

[0069] Example 46 Synthesis of Compound 46 ; Synthesis Scheme: ; Step 1: Synthesis of 2-cyclopropyl-4-fluoroaniline: 2-Cyclopropyl-4-fluoroaniline was prepared according to the procedure of Step 1 in Example 1. Characterization: LCMS m / z = 152.08 [M+1] + Step 2: Synthesis of N-(2-cyclopropyl-4-fluorophenyl)pyrimidin-2-amine ; To a reaction flask were added 2-bromopyrimidine (1.16 g, 7.28 mmol), 2-bromo-4-fluoroaniline (1.00 g, 6.61 mmol), p-toluenesulfonic acid (1.71 g, 9.92 mmol), and isopropanol (5 ml). The temperature was raised to 90 °C and the mixture was stirred for 8 hours. After the reaction was completed, it was cooled to room temperature, diluted with pure water, extracted with dichloromethane, concentrated, and purified by column chromatography to obtain N-(2-cyclopropyl-4-fluorophenyl)pyrimidin-2-amine as a light brown liquid (1.38 g, yield 91%, purity 94.6%). Characterization: LCMS m / z = 230.10 [M+1] + Step 3: Synthesis of N-(2-cyclopropyl-4-fluorophenyl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine N-(2-Cyclopropyl-4-fluorophenyl)-7-nitro-N-(pyrimidin-2-yl)benzo[c][1,2,5]oxadiazol-4-amine was prepared according to the procedure of Step 3 in Example 1. Characterization: LCMS m / z = 393.10 [M+1] +

[0070] Example 47 Synthesis of Compound 47 ; Synthesis of N-(2-cyclopropyl-4-fluorophenyl)-7-nitro-N-(thiazol-2-yl)benzo[c][1,2,5]oxadiazol-4-amine N-(2-Cyclopropyl-4-fluorophenyl)-7-nitro-N-(thiazol-2-yl)benzo[c][1,2,5]oxadiazol-4-amine was prepared according to the procedures of Steps 1 - 3 in Example 46. Characterization: LCMS m / z = 397.06 [M+1] +

[0071] Example 48 Synthesis of Compound 48 ; Synthesis of N-(2-cyclopropyl-4-fluorophenyl)-N-(1H-imidazol-2-yl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine Prepare N-(2-cyclopropyl-4-fluorophenyl)-N-(1H-imidazol-2-yl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine according to the steps of Step 1-3 in Example 46. Characterization: LCMS m / z = 381.10 [M+1] +

[0072] Example 49 Synthesis of Compound 49 ; Synthesis of 6-((2-cyclopropyl-4-fluorophenyl)(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)pyrimidine-2,4(1H,3H)-dione Prepare 6-((2-cyclopropyl-4-fluorophenyl)(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)pyrimidine-2,4(1H,3H)-dione according to the steps of Step 1-3 in Example 46. Characterization: 1 H NMR (400 MHz, DMSO) δ 10.95 (s, 1H), 10.58 (s, 1H), 8.68 (d, J = 7.7 Hz, 1H), 8.43 (s, 1H), 7.71 (d, J = 7.7 Hz, 1H), 7.05 (dd, J = 8.7, 5.6Hz, 1H), 6.85 (td, J = 8.5, 2.9 Hz, 1H), 6.61 (dd, J = 10.4, 2.9 Hz, 1H), 2.01-1.99 (m, 1H), 0.99 – 0.92 (m, 2H), 0.69-0.67 (m, 2H). LCMS m / z = 425.09 [M+1] +

[0073] Example 50 Synthesis of Compound 50 ; Synthesis of N-(2-cyclopropyl-4-fluorophenyl)-N-(1-methyl-1H-1,2,3-triazol-4-yl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine Prepare N-(2-cyclopropyl-4-fluorophenyl)-N-(1-methyl-1H-1,2,3-triazol-4-yl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine according to the steps of Step 1-3 in Example 46. Characterization: LCMS m / z = 396.11 [M+1] +

[0074] Example 51 Synthesis of Compound 51 ; Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-N-(5-methylthiazol-2-yl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine Prepare N-(2-Cyclopropyl-4-fluorophenyl)-N-(5-methylthiazol-2-yl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine according to the steps of steps 1-3 of Example 46. Characterization: LCMS m / z = 412.08 [M+1] +

[0075] Example 52 Synthesis of Compound 52 ; Synthesis of N-(5-Chlorothiazol-2-yl)-N-(2-cyclopropyl-4-fluorophenyl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine Prepare N-(5-Chlorothiazol-2-yl)-N-(2-cyclopropyl-4-fluorophenyl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine according to the steps of steps 1-3 of Example 46. Characterization: LCMS m / z = 432.03 [M+1] +

[0076] Example 53 Synthesis of Compound 53 ; Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-N-(4,5-dimethylthiazol-2-yl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine Prepare N-(2-Cyclopropyl-4-fluorophenyl)-N-(4,5-dimethylthiazol-2-yl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine according to the steps of steps 1-3 of Example 46. Characterization: LCMS m / z = 426.10 [M+1] +

[0077] Example 54 Synthesis of Compound 54 ; Synthesis of N-(4-Chlorothiazol-2-yl)-N-(2-cyclopropyl-4-fluorophenyl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine Prepare N-(4-Chlorothiazol-2-yl)-N-(2-cyclopropyl-4-fluorophenyl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine according to the steps of steps 1-3 of Example 46. Characterization: LCMS m / z = 432.03 [M+1] +

[0078] Example 55 Synthesis of Compound 55 ; Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-N-(2-methylthiazol-4-yl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine Prepare N-(2-Cyclopropyl-4-fluorophenyl)-N-(2-methylthiazol-4-yl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine according to the procedures of Steps 1-3 in Example 46. Characterization: LCMS m / z = 412.08 [M+1] +

[0079] Example 56 Synthesis of Compound 56 ; Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-N-(1,3-dimethyl-1H-pyrazol-4-yl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine Prepare N-(2-Cyclopropyl-4-fluorophenyl)-N-(1,3-dimethyl-1H-pyrazol-4-yl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine according to the procedures of Steps 1-3 in Example 46. Characterization: LCMS m / z = 409.13 [M+1] +

[0080] Example 57 Synthesis of Compound 57 ; Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-N-(4,5-dimethyloxazol-2-yl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine Prepare N-(2-Cyclopropyl-4-fluorophenyl)-N-(4,5-dimethyloxazol-2-yl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine according to the procedures of Steps 1-3 in Example 46. Characterization: 11H NMR (600 MHz, DMSO) δ 8.65 (d, J = 8.5 Hz, 1H), 7.45 (dd, J = 8.7, 5.4 Hz, 1H), 7.17 (td, J = 8.4, 2.9 Hz, 1H), 7.11 (d, J = 8.5 Hz, 1H), 7.01 (dd, J = 10.1, 2.8 Hz, 1H), 2.20 (s, 3H), 2.04 (s, 3H), 1.78 – 1.71 (m, 1H), 0.77 - 0.60 (m, 4H). LCMS m / z = 410.12 [M+1] +

[0081] Example 58 Synthesis of Compound 58 ; Synthesis of N-(4-Fluoro-2-methylphenyl)-7-nitro-N-(oxazol-2-yl)benzo[c][1,2,5]oxadiazol-4-amine Prepare N-(4-Fluoro-2-methylphenyl)-7-nitro-N-(oxazol-2-yl)benzo[c][1,2,5]oxadiazol-4-amine according to the steps of Steps 1 - 3 of Example 46. Characterization: LCMS m / z = 356.07 [M+1] +

[0082] Example 59 Synthesis of Compound 59 ; Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-N-(4-methylthiazol-2-yl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine Prepare N-(2-Cyclopropyl-4-fluorophenyl)-N-(4-methylthiazol-2-yl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine according to the steps of Steps 1 - 3 of Example 46. Characterization: LCMS m / z = 412.08 [M+1] +

[0083] Example 60 Synthesis of Compound 60 ; Synthesis of N-(2-Cyclopropyl-4-fluorophenyl)-N-(5-methyl-1,3,4-oxadiazol-2-yl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine Prepare N-(2-cyclopropyl-4-fluorophenyl)-N-(5-methyl-1,3,4-oxadiazol-2-yl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine according to the steps of Step 1-3 in Example 46. Characterization: LCMS m / z = 397.10 [M+1] +

[0084] Example 61 Synthesis of Compound 61 ; Synthesis of N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)-2-(trifluoromethyl)pyrimidine-5-carboxamide Prepare N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)-2-(trifluoromethyl)pyrimidine-5-carboxamide according to the steps of Step 1-3 in Example 1. Characterization: 1 H NMR (400 MHz, DMSO) δ 9.22 (s, 1H), 8.71 (d, J = 8.0 Hz, 1H), 7.63(dd, J = 11.2, 5.7 Hz, 1H), 7.10 (td, J = 8.4, 2.6 Hz, 1H), 6.85 (dd, J =10.1, 2.4 Hz, 1H), 2.04 – 1.93 (m, 1H), 0.91-0.95 (m, 1H), 0.73 – 0.64 (m,1H), 0.57 (d, J = 4.0 Hz, 1H). LCMS m / z = 489.09 [M+1] +

[0085] Example 62 Synthesis of Compound 62 ; Synthesis of 4-amino-N-(2-cyclopropyl-4-fluorophenyl)-2-methyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)pyrimidine-5-carboxamide Prepare 4-amino-N-(2-cyclopropyl-4-fluorophenyl)-2-methyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)pyrimidine-5-carboxamide according to the steps of Step 1-3 in Example 1. Characterization: 11H NMR (600 MHz, DMSO) δ 12.89 (s, 1H), 10.66 (s, 1H), 9.34 (s, 1H), 8.80 (dd, J = 22.3, 8.5 Hz, 2H), 7.36 (dd, J = 8.5, 5.7 Hz, 1H), 7.09 (td, J = 8.4, 2.8 Hz, 1H), 6.85 (dd, J = 10.3, 2.6 Hz, 1H), 2.74 (s, 3H), 2.09 – 2.01 (m, 1H), 0.93 (t, J = 6.4 Hz, 2H), 0.71 (d, J = 4.4 Hz, 2H). LCMS m / z = 450.12 [M+1] +

[0086] Example 63 Synthesis of Compound 63 ; Synthesis of 5-Amino-N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)pyridazine-4-carboxamide 5-Amino-N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)pyridazine-4-carboxamide was prepared according to the procedures of Steps 1 - 3 in Example 1. Characterization data are as follows: 1 1H NMR (400 MHz, DMSO) δ 10.97 (s, 1H), 8.51 (d, J = 8.8 Hz, 1H), 7.38 (dd, J = 8.7, 5.6 Hz, 1H), 7.15 (td, J = 8.4, 2.9 Hz, 1H), 6.92 (dd, J = 10.4, 2.9 Hz, 1H), 6.01 (d, J = 8.8 Hz, 1H), 1.98 – 1.89 (m, 1H), 0.91 – 0.85 (m, 2H), 0.77 – 0.71 (m, 2H). LCMS m / z = 436.11 [M+1] +

[0087] Example 64 Synthesis of Compound 64 ; Synthesis of 2-Amino-N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)-4-(trifluoromethyl)pyrimidine-5-carboxamide Prepare 2-amino-N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)-4-(trifluoromethyl)pyrimidine-5-carboxamide according to the steps of Step 1-3 in Example 1. Characterization: LCMS m / z = 504.10 [M+1] +

[0088] Example 65 Synthesis of Compound 65 ; Synthesis of 6-amino-N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)pyrazine-2-carboxamide Prepare 6-amino-N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)pyrazine-2-carboxamide according to the steps of Step 1-3 in Example 1. Characterization: LCMS m / z = 436.11 [M+1] +

[0089] Example 66 Synthesis of Compound 66 ; Synthesis Scheme: ; Synthesis of (R)-N-(4-fluoro-2-methylphenyl)-7-nitro-N-(oxetan-2-ylmethyl)benzo[c][1,2,5]oxadiazol-4-amine Prepare (R)-N-(4-fluoro-2-methylphenyl)-7-nitro-N-(oxetan-2-ylmethyl)benzo[c][1,2,5]oxadiazol-4-amine according to the steps of Step 2-3 in Example 46. Characterization: LCMS m / z = 359.11 [M+1] +

[0090] Example 67 Synthesis of Compound 67 ; Synthesis of 3-methoxy-4-(2,2,3,3-tetramethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopropane-1-carboxamido)benzoic acid Prepare 3-methoxy-4-(2,2,3,3-tetramethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopropane-1-carboxamido)benzoic acid according to the steps of Step 2-3 in Example 1. Characterization: LCMS m / z = 455.15 [M+1] +

[0091] Example 68 Synthesis of Compound 68 ; Synthesis of 3-Cyclopropyl-4-(2,2,3,3-tetramethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopropane-1-carboxamide)benzamide Prepare 3-cyclopropyl-4-(2,2,3,3-tetramethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopropane-1-carboxamide)benzamide according to the steps of Step 1-3 in Example 1. Characterization: LCMS m / z = 464.19 [M+1] +

[0092] Example 69 Synthesis of Compound 69 ; Synthesis of N-(4-Fluoro-2-methylphenyl)-3,3-dimethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)butanamide Prepare N-(4-fluoro-2-methylphenyl)-3,3-dimethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)butanamide according to the steps of Step 2-3 in Example 1. Characterization: LCMS m / z = 387.14 [M+1] +

[0093] Example 70 Synthesis of Compound 70 ; Synthesis of 4-Methyl-3-(2,2,3,3-tetramethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopropane-1-carboxamido)benzoic Acid Prepare 4-methyl-3-(2,2,3,3-tetramethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopropane-1-carboxamido)benzoic acid according to the steps of Step 2-3 in Example 1. Characterization: LCMS m / z = 389.15 [M+1] +

[0094] Example 71 Synthesis of Compound 71 ; Synthesis of 3-Methyl-4-(2,2,3,3-tetramethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopropane-1-carboxamido)benzoic Acid Prepare 3-methyl-4-(2,2,3,3-tetramethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopropane-1-carboxamido)benzoic acid according to the steps of Step 2-3 in Example 1. Characterization: LCMS m / z = 389.15 [M+1] +

[0095] Example 72 Synthesis of Compound 72 ; Synthesis of 3-cyclopropyl-4-(2,2,3,3-tetramethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopropane-1-carboxamido)benzoic acid Prepare 3-cyclopropyl-4-(2,2,3,3-tetramethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopropane-1-carboxamido)benzoic acid according to the steps of Step 1-3 in Example 1. Characterization: LCMS m / z = 465.17 [M+1] +

[0096] Example 73 Synthesis of Compound 73 ; Synthesis of N-(2-cyclopropyl-5-fluorophenyl)-2,2,3,3-tetramethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopropane-1-carboxamide Prepare N-(2-cyclopropyl-5-fluorophenyl)-2,2,3,3-tetramethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopropane-1-carboxamide according to the steps of Step 1-3 in Example 1. Characterization: LCMS m / z = 439.17 [M+1] +

[0097] Example 74 Synthesis of Compound 74 ; Synthesis of 2-cyclopropyl-4-fluoro-N-(4-fluoro-2-methylphenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide Prepare 2-cyclopropyl-4-fluoro-N-(4-fluoro-2-methylphenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamide according to the steps of Step 2-3 in Example 1. Characterization: LCMS m / z = 451.11 [M+1] +

[0098] Example 75 Synthesis of Compound 75 ; Synthesis of 4-Cyclopropyl-3-(2,2,3,3-tetramethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopropane-1-carboxamido)benzoic Acid Prepare 4-cyclopropyl-3-(2,2,3,3-tetramethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopropane-1-carboxamido)benzoic acid according to the steps of Step 1-3 in Example 1. Characterization: LCMS m / z = 465.17 [M+1] +

[0099] Example 76 Synthesis of Compound 76 ; Synthesis of 2-Fluoro-4-methyl-5-(2,2,3,3-tetramethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopropane-1-carboxamido)benzoic Acid Prepare 2-fluoro-4-methyl-5-(2,2,3,3-tetramethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)cyclopropane-1-carboxamido)benzoic acid according to the steps of Step 2-3 in Example 1. Characterization: LCMS m / z = 457.14 [M+1] +

[0100] Example 77 Synthesis of Compound 77 ; Synthesis of 3-(2,4-Dimethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamido)-4-methylbenzoic Acid Prepare 3-(2,4-dimethyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)benzamido)-4-methylbenzoic acid according to the steps of Step 2-3 in Example 1. Characterization: LCMS m / z = 447.12 [M+1] +

[0101] Example 78 Synthesis of Compound 78 ; Synthesis of Methyl (2-Cyclopropyl-4-fluorophenyl)(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)carbamate Prepare methyl (2-cyclopropyl-4-fluorophenyl)(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)carbamate according to the steps of Step 1-3 in Example 1. Characterization: LCMS m / z = 373.09 [M+1] +

[0102] Example 79 Synthesis of Compound 79 ; Synthesis of Ethyl (2-cyclopropyl-4-fluorophenyl)(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)carbamate Ethyl (2-cyclopropyl-4-fluorophenyl)(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)carbamate was prepared according to the steps of Steps 1-3 of Example 1. Characterization: LCMS m / z = 387.10 [M+1] +

[0103] Example 80 Synthesis of Compound 80 ; Synthesis of 2,5-Dichloro-N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)thiazole-4-carboxamide 2,5-Dichloro-N-(2-cyclopropyl-4-fluorophenyl)-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)thiazole-4-carboxamide was prepared according to the steps of Steps 1-3 of Example 1. Characterization: 1 H NMR (400 MHz, DMSO) δ 8.66 (d, J = 8.0 Hz, 1H), 7.40 (dd, J = 8.7,5.5 Hz, 1H), 7.32 (d, J = 8.0 Hz, 1H), 7.10 (td, J = 8.4, 2.9 Hz, 1H), 6.92(dd, J = 10.2, 2.9 Hz, 1H), 1.89 – 1.80 (m, 1H), 0.85 (tt, J = 6.3, 4.5 Hz,1H), 0.78 (dt, J = 10.6, 4.3 Hz, 1H), 0.63 – 0.55 (m, 1H), 0.51 (dq, J =10.8, 4.9 Hz, 1H). LCMS m / z = 493.98 [M+1] +

[0104] Example 81 Synthesis of Compound 81 ; N-(2-Cyclopropyl-4-fluorophenyl)-2-hydroxy-6-methyl-N-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)isonicotinamide was prepared according to the steps of Steps 1-3 of Example 1.

[0105] Characterization: 1 1H NMR (400 MHz, DMSO) δ 8.66 (d, J = 8.0 Hz, 1H), 7.51 (dd, J = 8.5,5.5 Hz, 1H), 7.36 (d, J = 8.0 Hz, 1H), 7.08 (td, J = 8.3, 2.5 Hz, 1H), 6.87(dd, J = 10.1, 2.3 Hz, 1H), 6.27 (s, 1H), 6.14 (s, 1H), 2.12 (s, 3H), 1.86-1.82 (m, 1H), 0.96-0.93 (m, 1H), 0.81 – 0.73 (m, 1H), 0.56-0.54 (m, 2H).LCMSm / z=450.11[M+1] +

[0106] Example 82 The inhibitory activity of the disclosed compound of the present invention against human Burkitt's lymphoma cells (Daudi cells) and proliferation.

[0107] Daudi cells (Cell Resource Center, Institute of Basic Medicine, Chinese Academy of Medical Sciences) were cultured in complete RPMI Medium 1640 basic (C11875500BT) containing 10% fetal bovine serum. On the first day of the experiment, Daudi cells were seeded in a 96-well plate at a density of 10,000 cells / well with 100 μL of cell suspension per well using RPMI 1640 medium containing 10% fetal bovine serum. 300 μL of PBS was added to the side wells. The plate was placed in a cell culture incubator at 37 °C with 5% CO 2 overnight. On the second day, 100 μL of the test compound at different concentrations prepared with the medium was added to each well. The final concentrations of the compound were set at 7 concentration points: 25 μM, 10 μM, 7.5 μM, 5 μM, 3.75 μM, 2.5 μM, and 1 μM (adjusted as needed according to the experimental purpose). A blank control was set, and the plate was placed in a cell culture incubator at 37 °C with 5% CO 2 for 24 hours. On the third day, the 96-well plate was taken out, and the medium in the experimental wells and the blank control wells was completely aspirated. CCK-8 was prepared at a ratio of 10:1 (RPMI 1640: Cell Counting Kit-8, CK04), and 100 μL was added to each well. The plate was placed in a cell culture incubator at 37 °C with 5% CO 2After incubation in a cell incubator for 2 h, the luminescence signal value was read using a multi-label microplate reader (BioTek, Synergy H1), and the IC of the inhibitory activity of the compound was calculated using Graphpad Prism software based on the concentration of the compound and the luminescence signal value. 50 value.

[0108] The IC of the inhibitory effect of the compounds disclosed in the present invention on the proliferation of Daudi cells 50 values are shown in Table 1 below.

[0109] Table 1 IC of the inhibitory effect of the compounds disclosed in the present invention on the proliferation of Daudi cells 50 values (μM) Serial number IC50 Serial number IC50 Serial number IC50 Serial number IC50 Compound 1 3.169 Compound 2 4.843 Compound 3 66.121 Compound 4 7.108 Compound 5 0.782 Compound 6 3.892 Compound 7 4.728 Compound 8 0.026 Compound 9 4.827 Compound 10 0.174 Compound 11 5.839 Compound 12 12.839 Compound 13 28.337 Compound 14 7.822 Compound 15 0.468 Compound 16 0.047 Compound 17 44.282 Compound 18 16.892 Compound 19 6.821 Compound 20 5.927 Compound 21 5.892 Compound 22 38.921 Compound 23 1.836 Compound 24 0.468 Compound 25 0.728 Compound 26 6.821 Compound 27 33.921 Compound 28 3.731 Compound 29 6.302 Compound 30 0.563 Compound 31 7.839 Compound 32 58.392 Compound 33 7.382 Compound 34 5.382 Compound 35 6.934 Compound 36 0.842 Compound 37 15.621 Compound 38 29.483 Compound 39 7.389 Compound 40 28.893 Compound 41 0.362 Compound 42 6.871 Compound 43 0.284 Compound 44 5.839 Compound 45 6.389 Compound 46 0.837 Compound 47 4.785 Compound 48 6.832 Compound 49 2.893 Compound 50 6.832 Compound 51 8.932 Compound 52 8.921 Compound 53 6.829 Compound 54 0.983 Compound 55 0.883 Compound 56 68.236 Compound 57 3.841 Compound 58 2.715 Compound 59 6.391 Compound 60 16.378 Compound 61 6.381 Compound 62 16.381 Compound 63 28.392 Compound 64 0.047 Compound 65 8.492 Compound 66 1.382 Compound 67 6.932 Compound 68 6.921 Compound 69 16.29 Compound 70 2.832 Compound 71 5.291 Compound 72 4.291 Compound 73 2.142 Compound 74 0.372 Compound 75 0.472 Compound 76 18.38 Compound 77 6.382 Compound 78 5.281 Compound 79 8.392 Compound 80 22.392 Compound 81 9.392 The above results indicate that the compounds provided by the present invention have a good inhibitory effect on the proliferation of Daudi cells.

[0110] Example 83 The inhibitory activity of the compounds disclosed in the present invention on the proliferation of human colorectal cancer cells (A2780 cells) is shown in Table 2 below.

[0111] Referring to the experimental procedure of Example 82, the inhibitory effect of the compounds disclosed in the present invention on the proliferation of A2780 cells was determined. Among them, A2780 cells (Cell Resource Center, Institute of Basic Medicine, Chinese Academy of Medical Sciences) were cultured in RPMI Medium 1640 basic (C11875500BT) complete medium containing 10% fetal bovine serum.

[0112] The IC of the inhibitory effect of the compounds disclosed in the present invention on the proliferation of A2780 cells 50 values are shown in Table 2 below.

[0113] Table 2 IC of the inhibitory effect of the compounds disclosed in the present invention on the proliferation of A2780 cells 50 values (μM) Serial number IC50 Serial number IC50 Serial number IC50 Serial number IC50 Compound 1 0.742 Compound 2 6.382 Compound 3 3.842 Compound 4 12.483 Compound 5 6.382 Compound 6 5.382 Compound 7 0.382 Compound 8 0.473 Compound 9 4.282 Compound 10 0.989 Compound 11 1.483 Compound 12 28.382 Compound 13 8.378 Compound 14 9.387 Compound 15 0.373 Compound 16 0.854 Compound 17 36.483 Compound 18 18.593 Compound 19 5.839 Compound 20 6.393 Compound 21 19.483 Compound 22 5.932 Compound 23 5.382 Compound 24 7.392 Compound 25 5.433 Compound 26 0.954 Compound 27 0.467 Compound 28 5.383 Compound 29 0.854 Compound 30 0.542 Compound 31 8.383 Compound 32 18.372 Compound 33 5.822 Compound 34 9.320 Compound 35 11.392 Compound 36 0.092 Compound 37 6.861 Compound 38 18.963 Compound 39 5.862 Compound 40 7.825 Compound 41 0.854 Compound 42 0.863 Compound 43 0.527 Compound 44 5.865 Compound 45 6.574 Compound 46 0.754 Compound 47 7.953 Compound 48 3.876 Compound 49 19.853 Compound 50 5.876 Compound 51 7.382 Compound 52 3.982 Compound 53 6.937 Compound 54 5.839 Compound 55 5.836 Compound 56 6.893 Compound 57 5.882 Compound 58 16.922 Compound 59 6.832 Compound 60 18.392 Compound 61 29.483 Compound 62 48.683 Compound 63 67.382 Compound 64 0.846 Compound 65 17.983 Compound 66 4.978 Compound 67 6.983 Compound 68 7.831 Compound 69 11.832 Compound 70 8.832 Compound 71 15.842 Compound 72 5.832 Compound 73 29.842 Compound 74 0.902 Compound 75 7.832 Compound 76 4.832 Compound 77 7.832 Compound 78 3.985 Compound 79 18.393 Compound 80 19.348 Compound 81 2.483 The above results indicate that the compounds provided by the present invention have a good inhibitory effect on the proliferation of A2780 cells.

[0114] Example 84 The inhibitory activity of the compounds disclosed in the present invention on the proliferation of human colorectal cancer cells (COLO320DM cells) is shown in Table 3 below.

[0115] Referring to the experimental procedure of Example 82, the inhibitory effect of the disclosed compounds of the present invention on the proliferation of COLO320DM cells was determined. Among them, COLO320DM cells (Cell Resource Center, Institute of Basic Medicine, Chinese Academy of Medical Sciences) were cultured in complete medium of RPMI Medium 1640 basic (C11875500BT) containing 10% fetal bovine serum.

[0116] The inhibitory IC of the disclosed compounds of the present invention on the proliferation of COLO320DM cells 50 values are shown in Table 3 below. A = compound IC 50 less than or equal to 0.1 μM, B = compound IC 50 greater than 0.1 μM and less than or equal to 1 μM, C = compound IC 50 greater than 1 μM and less than or equal to 10 μM, D = compound IC 50 greater than 10 μM.

[0117] Table 3 The inhibitory IC of the disclosed compounds of the present invention on the proliferation of COLO320DM cells 50 values (μM) Serial number IC50 Serial number IC50 Serial number IC50 Serial number IC50 Compound 1 1.849 Compound 2 0.532 Compound 3 15.982 Compound 4 11.831 Compound 5 5.282 Compound 6 6.382 Compound 7 0.637 Compound 8 0.842 Compound 9 2.473 Compound 10 7.348 Compound 11 5.392 Compound 12 28.493 Compound 13 6.382 Compound 14 3.752 Compound 15 9.487 Compound 16 0.462 Compound 17 2.472 Compound 18 184.93 Compound 19 1.482 Compound 20 7.348 Compound 21 0.372 Compound 22 0.662 Compound 23 0.473 Compound 24 0.742 Compound 25 7.492 Compound 26 0.832 Compound 27 0.732 Compound 28 4.621 Compound 29 18.392 Compound 30 3.921 Compound 31 3.842 Compound 32 6.372 Compound 33 37.323 Compound 34 15.721 Compound 35 8.931 Compound 36 0.037 Compound 37 4.928 Compound 38 5.923 Compound 39 0.932 Compound 40 0.462 Compound 41 0.483 Compound 42 0.573 Compound 43 2.472 Compound 44 7.482 Compound 45 0.383 Compound 46 6.483 Compound 47 0.783 Compound 48 0.458 Compound 49 8.482 Compound 50 6.387 Compound 51 5.383 Compound 52 6.483 Compound 53 0.593 Compound 54 0.573 Compound 55 7.438 Compound 56 18.38 Compound 57 2.483 Compound 58 5.483 Compound 59 6.493 Compound 60 11.482 Compound 61 0.942 Compound 62 28.503 Compound 63 18.583 Compound 64 0.489 Compound 65 17.483 Compound 66 0.993 Compound 67 0.463 Compound 68 6.382 Compound 69 7.483 Compound 70 6.833 Compound 71 6.483 Compound 72 7.832 Compound 73 19.842 Compound 74 8.483 Compound 75 7.483 Compound 76 8.483 Compound 77 5.372 Compound 78 3.942 Compound 79 19.458 Compound 80 4.382 Compound 81 8.437 The above results show that the compounds provided by the present invention have good inhibitory effects on the proliferation of COLO320DM cells.

[0118] Example 85 The inhibitory activity of the disclosed compounds of the present invention on the proliferation of human osteosarcoma cells (SJSA-1 cells) is shown in Table 4 below.

[0119] Referring to the experimental procedure of Example 82, the inhibitory effect of the disclosed compounds of the present invention on the proliferation of SJSA-1 cells was determined. Among them, SJSA-1 cells (Cell Resource Center, Institute of Basic Medicine, Chinese Academy of Medical Sciences) were cultured in complete medium of RPMI Medium 1640 basic (C11875500BT) containing 10% fetal bovine serum.

[0120] The inhibitory IC of the disclosed compounds of the present invention on the proliferation of SJSA-1 cells 50 values are shown in Table 4 below.

[0121] Table 4 The inhibitory IC of the disclosed compounds of the present invention on the proliferation of SJSA-1 cells 50 values Serial number IC50 Serial number IC50 Serial number IC50 Serial number IC50 Compound 1 7.392 Compound 2 0.921 Compound 3 7.382 Compound 4 10.482 Compound 5 6.392 Compound 6 8.331 Compound 7 4.881 Compound 8 0.856 Compound 9 7.326 Compound 10 10.731 Compound 11 0.063 Compound 12 11.724 Compound 13 7.392 Compound 14 6.372 Compound 15 3.801 Compound 16 0.744 Compound 17 6.302 Compound 18 13.482 Compound 19 5.184 Compound 20 5.717 Compound 21 0.583 Compound 22 7.382 Compound 23 8.312 Compound 24 0.884 Compound 25 8.382 Compound 26 0.462 Compound 27 0.642 Compound 28 0.718 Compound 29 19.372 Compound 30 7.881 Compound 31 4.281 Compound 32 6.972 Compound 33 18.143 Compound 34 3.582 Compound 35 6.385 Compound 36 0.286 Compound 37 8.372 Compound 38 6.382 Compound 39 0.063 Compound 40 0.174 Compound 41 6.382 Compound 42 0.763 Compound 43 3.750 Compound 44 6.876 Compound 45 5.282 Compound 46 9.371 Compound 47 0.483 Compound 48 0.882 Compound 49 6.353 Compound 50 7.533 Compound 51 8.482 Compound 52 7.372 Compound 53 0.372 Compound 54 0.528 Compound 55 4.722 Compound 56 12.467 Compound 57 9.362 Compound 58 8.819 Compound 59 10.48 Compound 60 10.472 Compound 61 8.438 Compound 62 7.742 Compound 63 16.38 Compound 64 0.362 Compound 65 19.328 Compound 66 0.642 Compound 67 0.437 Compound 68 3.841 Compound 69 6.472 Compound 70 0.921 Compound 71 9.382 Compound 72 9.371 Compound 73 10.246 Compound 74 0.682 Compound 75 6.428 Compound 76 4.817 Compound 77 4.821 Compound 78 9.376 Compound 79 3.471 Compound 80 7.300 Compound 81 2.472 The above results show that the compounds provided by the present invention have good inhibitory effects on the proliferation of SJSA-1 cells.

[0122] Example 86 The inhibitory activity of the disclosed compounds of the present invention on the proliferation of human myelomonocytic leukemia cells (MV4-11 cells) is shown in Table 5 below.

[0123] Referring to the experimental procedure of Example 82, the inhibitory effect of the disclosed compounds of the present invention on the proliferation of MV4-11 cells was determined. Among them, MV4-11 cells (Cell Resource Center, Institute of Basic Medicine, Chinese Academy of Medical Sciences) were cultured in RPMI Medium 1640 basic (C11875500BT) complete medium containing 10% fetal bovine serum.

[0124] The inhibitory IC 50 value of the disclosed compounds of the present invention on the proliferation of MV4-11 cells is shown in Table 5 below.

[0125] Table 5 The inhibitory IC 50 value Serial number IC50 Serial number IC50 Serial number IC50 Serial number IC50 Compound 1 5.832 Compound 2 0.732 Compound 3 17.389 Compound 4 28.472 Compound 5 0.742 Compound 6 7.348 Compound 7 5.382 Compound 8 0.842 Compound 9 6.382 Compound 10 0.164 Compound 11 8.182 Compound 12 6.831 Compound 13 7.831 Compound 14 5.831 Compound 15 0.732 Compound 16 0.027 Compound 17 19.482 Compound 18 28.372 Compound 19 12.842 Compound 20 7.830 Compound 21 6.382 Compound 22 13.482 Compound 23 6.382 Compound 24 4.281 Compound 25 0.482 Compound 26 3.842 Compound 27 48.392 Compound 28 7.821 Compound 29 0.642 Compound 30 0.078 Compound 31 6.382 Compound 32 4.832 Compound 33 5.281 Compound 34 11.382 Compound 35 1.482 Compound 36 1.472 Compound 37 16.382 Compound 38 11.372 Compound 39 4.272 Compound 40 5.324 Compound 41 0.482 Compound 42 6.382 Compound 43 0.742 Compound 44 6.482 Compound 45 0.472 Compound 46 0.984 Compound 47 6.382 Compound 48 2.482 Compound 49 6.821 Compound 50 5.382 Compound 51 0.482 Compound 52 0.572 Compound 53 7.382 Compound 54 0.762 Compound 55 0.472 Compound 56 4.382 Compound 57 8.492 Compound 58 8.498 Compound 59 19.481 Compound 60 17.381 Compound 61 5.382 Compound 62 10.482 Compound 63 5.382 Compound 64 0.931 Compound 65 9.482 Compound 66 0.172 Compound 67 7.480 Compound 68 0.462 Compound 69 7.825 Compound 70 4.731 Compound 71 8.612 Compound 72 5.281 Compound 73 6.382 Compound 74 0.462 Compound 75 0.562 Compound 76 10.382 Compound 77 1.482 Compound 78 5.382 Compound 79 0.792 Compound 80 6.382 Compound 81 7.382 The above results indicate that the compounds provided by the present invention have a good inhibitory effect on the proliferation of MV4-11 cells.

[0126] Example 87 The inhibitory activity of the disclosed compounds of the present invention on the expression of MYC in Daudi cells 1. Cell culture: Take the Daudi cells cryopreserved in the liquid nitrogen tank and resuscitate and culture them until the logarithmic growth phase. On the first day of the experiment, use RPMI 1640 medium containing 10% fetal bovine serum to inoculate Daudi cells at a density of 2×10^5 cells / well in a 6-well plate, with 1 mL of cell suspension in each well, and place it in a cell culture incubator at 37°C and 5% CO 2 overnight.

[0127] 2. Drug treatment Dose-effect: Weigh an appropriate amount of the compound and dissolve it in DMSO solvent, vortex and mix well to prepare a stock solution of 12 mM. Add the drug to act on Daudi cells respectively. The solvent control group is only treated with DMSO, and the drug treatment groups are respectively added with the corresponding compounds for treatment. The final concentration of its action is set to 12 μM, 6 μM, 3 μM, 1.5 μM and 0.75 μM, and the cells are collected after 12 h.

[0128] Time-effect: Weigh an appropriate amount of the compound and dissolve it in DMSO solvent, vortex and mix well to prepare a stock solution of 7.5 mM. Add the drug to act on Daudi cells respectively. The solvent control group is only treated with DMSO, and the drug treatment groups are respectively added with the corresponding compounds for treatment. The final concentration of its action is set to 7.5 μM (the inhibitory rate of activity screening reaches more than 90%), and the cells are collected after 0 h, 3 h, 6 h, 9 h, 12 h and 24 h.

[0129] 3. Cell collection After treating the cells with the compound and the solvent control group cells with 0.25% trypsin digestion, centrifuge again at 3000 rpm for 5 min at 4°C in 1 mL of RPMI 1640 complete medium, and completely discard the PBS after centrifugation.

[0130] 4. Cell lysis and protein collection Add an appropriate amount of lysis buffer to the collected cells, add protease inhibitor cocktail (P6730, 100×) and protein phosphatase inhibitor mixture (P1260, 100×) to the lysis buffer, pipette 20 times with a 1 mL pipette, lyse on ice for 30 min (oscillate once every 5 - 10 min), centrifuge at 12000 rpm for 30 min, and take the supernatant.

[0131] 5. Protein concentration determination by BCA method Prepare standard proteins with gradient concentrations, take 10 μL and put it in a 96 - well plate, and make two replicates for each standard. Take 2 μL of protein lysate and dissolve it in 8 μL of lysis buffer, and make two replicates for each sample. Add 100 μL of BCA working solution (Solution A: Solution B = 50:1) to each well, incubate at 37°C for 30 min. Measure the absorbance at 562 nm. Calculate the protein concentration of each sample and calculate the volume of protein lysate required for 10 μg.

[0132] 6. Detection of protein expression level by Western blot Take the protein lysate, add a constant volume and then add an appropriate amount of 5× protein loading buffer, denature at 98°C for 10 min. Use a 10% precast gel, electrophorese at 80 V for 30 min and then adjust to 120 V. When the sample electrophoreses to the appropriate position, cut off the stacking gel, transfer the protein on the separating gel to the PVDF membrane by wet transfer method, 200 mA, 2 h. Place the PVDF membrane in 5% skim milk and block at room temperature for 1 h. Cut the PVDF membrane according to the molecular weight of the protein Marker, place it in MYC and GAPDH antibodies respectively, and incubate overnight at 4°C. The next day, place the PVDF membrane in TBST, wash it by shaking 3 times, 10 min each time. Place the PVDF membrane in the corresponding secondary antibodies (goat anti - rabbit and goat anti - mouse) of MYC and GAPDH antibodies, incubate by shaking at room temperature for 2 h. After washing 3 times in TBST by shaking, add ECL luminescent solution and perform luminescence imaging in the Tanon 5200 fully automatic chemiluminescence image analysis system.

[0133] The experimental results obtained are shown in Figure 1 , Table 6 and Table 7.

[0134] Table 6 Dose - dependent degradation rate of the disclosed compound of the present invention on MYC expression in Daudi cells Serial number 12 μM 6 μM 3 μM 1.5 μM 0.75 μM Compound 8 96.38% 88.47% 62.47% 23.17% 9.27% Compound 11 91.47% 43.63% 15.18% 8.18% 9.27% Compound 24 87.27% 82.74% 80.36% 65.83% 26.81% Compound 28 87.18% 84.18% 77.82% 67.29% 7.29% Compound 70 95.10% 58.92% 35.19% 12.93% 4.91% Compound 72 96.01% 89.38% 82.19% 73.93% 67.15% Compound 75 98.05% 97.18% 96.99% 74.10% 48.94% Compound 81 91.29% 89.19% 81.10% 61.23% 43.90% Table 7 Time-dependent degradation rate of the compounds disclosed in the present invention on MYC expression in Daudi cells Serial number 24 h 12 h 9 h 6 h 3 h Compound 8 98.26% 95.37% 92.48% 88.28% 78.19% Compound 11 77.95% 63.19% 55.14% 53.82% 42.91% Compound 24 96.12% 82.19% 78.63% 62.77% 51.38% Compound 28 97.11% 82.14% 74.88% 69.18% 60.14% Compound 70 87.38% 89.28% 79.15% 76.92% 62.19% Compound 72 94.18% 90.05% 83.18% 79.37% 55.20% Compound 75 92.18% 85.19% 73.58% 1.47% 0.58% Compound 81 97.19% 81.29% 72.11% 65.28% 44.27% It shows that the compounds provided by the present invention have good degradation activity on MYC protein in Daudi cells.

[0135] In vivo anti-tumor inhibitory activity of continuous treatment with Compound 88 1. Tumor model construction Daudi cells (Cell Resource Center, Institute of Basic Medicine, Chinese Academy of Medical Sciences) were cultured in complete medium of RPMI Medium 1640 basic (C11875500BT) containing 10% fetal bovine serum. When the cells grew to 80-90% confluence and were passaged to a sufficient number, they were inoculated subcutaneously into BALB / C nude mice (purchased from Beijing Huafukang Biotechnology Co., Ltd.) at a density of 1×10^6 cells / mouse.

[0136] 2. Experimental animals BALB / C nude mice, 5-6 weeks old, weighing 18-20 g, were purchased from Beijing Huafukang Biotechnology Co., Ltd. The mice were housed in an independent ventilated cage (IVC) system for small animals at a temperature of 20-25°C and a humidity of 40%-70%, with 12 hours of light and 12 hours of darkness per day. Four nude mice were housed in each cage, and they could freely eat and drink. The feed was sterilized by ultraviolet irradiation, and the water was autoclaved. After three days of feeding, the mice were grouped for experiments, and Daudi cells were directly inoculated subcutaneously under the right axilla of the nude mice.

[0137] 3. Grouping and dosing regimen design When the average tumor volume reached 200 mm 3 , the mice were randomly grouped according to tumor volume and body weight, and the grouped mice were marked. They were divided into 3 groups, with 5 mice in each group: vehicle group (Vehicle), Compound 8 (10 mg / kg), and Compound 8 (15 mg / kg). Vehicle: 10% DMSO + 55% PEG400 + 35% (20% β-cyclodextrin - Saline). Administered intraperitoneally. Tumor size was recorded with vernier calipers.

[0138] 4. Statistical analysis The tumor volume was calculated using the following formula: length × width × width / 2. Analysis was performed using GraphPad Prism 8.0 software, and the significance was set at P < 0.05. All quantitative data were expressed as mean ± SEM. Comparison between two unpaired groups was evaluated using parametric Student's t-test or non-parametric Mann-Whitney U test. If the data were normally distributed, one-way analysis of variance (ANOVA) was used to test for differences between groups. If the data were not normally distributed, the Kruskal-Wallis test was used to test for differences between groups. The Shapiro-Wilk test was used to test the normality of the data. The Levene test was used to test for homogeneity of variance. Unless otherwise stated, all experiments used at least three individual mice or samples.

[0139] The results of the inhibition of tumor growth by Compound 8 within 30 days are shown in Figure 2 , indicating that Compound 8 of the present invention has good in vivo anti-tumor activity.

[0140] Example 89 In Vivo Anti-Tumor Inhibitory Activity of Compound 28 after Stopping Administration after Short-Term Treatment Referring to the experimental procedure of Example 88, the in vivo anti-tumor inhibitory activity of the compounds disclosed in the present invention after stopping administration after short-term treatment was determined. When the average tumor volume of nude mice subcutaneously inoculated with Daudi cells reached 200 mm 3 , they were randomly grouped according to tumor volume and body weight, and the grouped mice were marked and divided into 2 groups, with 4 mice in each group: the vehicle group (Vehicle) and Compound 28 (15 mg / kg). Vehicle: 10% DMSO + 55% PEG400 + 35% (20% β-cyclodextrin-Saline). Intraperitoneal administration was continued until the control tumors grew to 1500 mm 3 , and then the administration was stopped. The tumor size was recorded with vernier calipers.

[0141] The results of the inhibition of tumor growth by Compound 28 within 50 days are shown in Figure 3 , indicating that Compound 28 of the present invention not only has good in vivo anti-tumor activity during treatment, but also has a long-term tumor inhibitory effect after early treatment and stopping administration.

[0142] Example 90 In Vivo Anti-Tumor Inhibitory Activity of Compound 28 in Combination with Doxorubicin Referring to the experimental procedure of Example 88, the in vivo anti-tumor inhibitory activity of Compound 30 of the present invention in combination with doxorubicin was determined. When the average tumor volume of nude mice subcutaneously inoculated with Daudi cells reached 200 mm 3At that time, random grouping was performed according to tumor volume and body weight, and the grouped mice were marked and divided into 4 groups, with 4 mice in each group: the vehicle group (Vehicle), the doxorubicin group (4 mg / kg), the compound 28 group (15 mg / kg), and the combination group of doxorubicin (4 mg / kg) & compound 28 (15 mg / kg). Solvent: 10% DMSO + 55% PEG400 + 35% (20% β-cyclodextrin - Saline). A vernier caliper was used to record the tumor size.

[0143] The results of the inhibition of tumor growth by compound 28 and the first-line lymphoma drug doxorubicin within 30 days are shown in Figure 4 , which indicates that the compound 28 of the present invention significantly reduces the tumor growth in mice, and has better anti-tumor activity in combination with doxorubicin.

[0144] In summary, the present invention provides a compound capable of targeted degradation of MYC. After entering the patient's body, the compound can target and degrade MYC, thereby effectively inhibiting the proliferation of tumor cells such as lymphoma, osteosarcoma, and cancer cells such as colorectal cancer, and having good anti-tumor and anti-cancer effects; the compound can also inhibit the proliferation of myelomonocytic leukemia cells in the patient's body, providing a new treatment plan for the treatment of myelomonocytic leukemia; the compound has a synergistic effect when combined with doxorubicin and has better anti-tumor activity, and has broad application prospects in the field of medicine.

[0145] The technical features of the above-described embodiments can be combined arbitrarily. For the sake of brevity of description, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, it should be considered as the scope recorded in this specification.

[0146] The above-described embodiments only represent several implementation manners of the present invention, and their descriptions are relatively specific and detailed, but they should not be construed as limiting the scope of the invention patent. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can be made, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the present invention patent should be subject to the appended claims.

Claims

1. A compound capable of targeting and degrading MYC, characterized in that: The structure of the compound is: ； Among them, R 1 is selected from acetyl, o-fluorobenzoyl, m-fluorobenzoyl, p-fluorobenzoyl, 2,6-difluorobenzoyl, cyclohexanecarbonyl, cyclopentanecarbonyl, 2-(piperidin-1-yl)acetyl, 2-(4-methylpiperazin-1-yl)acetyl, cyclopropanecarbonyl, cyclobutanecarbonyl, 2,2,3,3-tetramethylcyclopropane-1-carbonyl, 2-morpholinoacetyl, 3-methylbutyryl, pyrimidin-2-yl, 3,3-dimethylbutyryl, 3,5-difluorobenzoyl, hydrogen, p-carboxybenzoyl, 3-hydroxy-3-methylbutyryl, thiazole-2-carbonyl, 3-methyl-2-((methylformyl)amino)butyryl, 2-amino-pyrimidine-5-carbonyl, 3-amino-3-methylbutyryl, 3,4-dimethyl Oxybenzoyl, 3,4,5-trimethoxyphenylpropionyl, 4-amino-5-(ethylsulfonyl)-benzoyl, 2-(4-(4-methylpiperazin-1-yl)piperidinyl)acetyl, 4-hydroxy-2,6-dimethylbenzoyl, 4-bromothiazole-2-carbonyl, 4-cyclopropylthiazole-2-carbonyl, 4,5-dimethylthiazole-2-carbonyl, 5-methylthiazole-2-carbonyl, 2-aminopyrimidine-5-carbonyl, 4-hydroxy-2-methylbenzoyl, 4-amino-2-methylbenzoyl, 4-amino-2-methoxybenzoyl, 3-methyl-1H-pyrazole-4-carbonyl, 3,5-dimethyl-1H-pyrazole-4-carbonyl, 3-methyl-2-((tert-butyloxycarbonyl)amino)butyramide, 4-amino-4-methyl-pentanoyl, 4-amino-3,5-dimethylbenzoyl, 2-aminoacetyl, 4-amino-5-chloro-2-methoxyphenylpropanoyl, 4-amino-5-chloro-2,3-dihydrobenzofuran-7-carbonyl, 7-methoxy-4-oxo-1,4-dihydroquinoline-6-carbonyl, 4-amino-5-chloro-2-ethoxyphenylpropanoyl, 5-chloro-2-methoxy-4-(methylamino)benzoyl, 4-amino-2,3,5,6-tetrafluorobenzoyl, -2,5-dimethoxy-3-nitrobenzoyl, oxazole-4-carbonyl, 3,5-diamino-6-chloropyrazine-2-carbonyl, oxazole-4-carboxylic acid, 1H-imidazole-2-carbonyl, pyrimidine- 2-yl, thiazol-2-yl, 1H-imidazol-2-yl, 2,4 (1H, 3H)-dicarbonyl pyrimidin-6-yl, 1-methyl-1H-1,2,3-triazol-4-yl, 5-methylthiazol-4-yl, 3-methylthiazol-2-yl, 5-chlorothiazol-2-yl, 4,5-dimethylthiazol-2-yl, 4-chlorothiazol-2-yl, 2-methylthiazol-4-yl, 1,3-dimethyl-1H-pyrazol-4-yl, 5-phenyloxazol-2-yl, 4,5-dimethyloxazol-2-yl, 5-methylformylthiazol-4-yl, 5-thiazole-4-carboxylic acid, oxazol-2-yl, 4-methylthiazol-2-yl, 5-methyl-1,3,4-oxadiazol-2-yl, 5-phenyl-1,3,4-oxadiazol-2-yl, oxadiazol-5-carboxylic acid ethyl ester-2-yl, 2-(trifluoromethyl)pyrimidine-5-carbonyl, 4-amino-2-methylpyrimidine-5-carbonyl, 5-amino-pyridazine-4-carbonyl, 2-amino-4-(trifluoromethyl)pyrimidine-5-carbonyl, 6-amino-pyrazine-2-carbonyl, 3-amino-2-methoxy-isonicotinyl, oxadiazol-4-yl, 2,2,3,3-tetramethylcyclopropane-1-carbonyl, 3,3-dimethylbutyryl, 3,3-dimethylbutyryl, 2-cyclopropyl-4-fluorobenzene-1-carbonyl, 2-fluorobenzene-1-carbonyl, 3,3-dimethyl-butyryl, 2-(Methoxycarbonyl)aminobutyryl, 2,4-dimethylbenzoyl, 2-((3-aminopyrazin-2-yl)thio)acetyl, 4-((tert-butoxycarbonyl)amino)benzoyl, 4-amino-5-(ethylsulfonyl)-2-methoxyphenylpropanoyl, methyl formate-1-yl, ethyl formate-1-yl, 2,5-dichloro-thiazole-4-carbonyl, 5-fluoro-2-hydroxy-nicotinoyl, 6-hydroxy-2-methyl-nicotinoyl, 5-oxo-4,5-dihydro-1H-1,2,4-triazole-3-carbonyl, 2-hydroxy-6-methylisonicotinyl, 1,2,5-oxadiazole-3-carbonitrile-6-yl; R 2 Selected from 2-cyclopropyl-4-fluorophenyl, 2-cyclopentyl-4-fluorophenyl, 5-fluoro-4'-methyl-[1,1'-biphenyl]-2-yl, 2-cyclohexyl-4-fluorophenyl, 2-hydroxy-4-fluorophenyl, 2-methyl-4-fluorophenyl, 2-methoxy-4-carboxyphenyl, 2-cyclopropyl-4-formylphenyl, 2-methyl-5-carboxyphenyl, neopentyl, 2-cyclohexyl-4-formylphenyl, 2-cyclohexyl-5-carboxyphenyl, 2-methyl-4-fluoro-5-carboxyphenyl, 5-fluoro-4'-carboxy-[1,1'-biphenyl]-2-yl.

2. A compound capable of targeting and degrading MYC and its application, characterized in that: The drug comprises the compound as claimed in claim 1.

3. The compound capable of targeted degradation of MYC and its use as claimed in claim 2, characterized in that: The drug also includes pharmaceutically acceptable excipients, which are pharmaceutically acceptable salts, excipients or carriers.

4. The compound capable of targeted degradation of MYC and its use as claimed in claim 3, characterized in that: The carrier is any one or more of a filler, a wetting agent, a binder, a disintegrant or a lubricant.

5. The compound capable of targeted degradation of MYC and its use as claimed in claim 4, characterized in that: The pharmaceutical composition is in the form of any one or more of aqueous infusion, powder, lotion, tincture, oil, emulsion, ointment, plaster or aerosol.

6. Use of the compound capable of targeted degradation of MYC as claimed in claim 1 in the preparation of anti-tumor drugs.

7. Use of the compound capable of targeted degradation of MYC as claimed in claim 1 in the preparation of anticancer drugs.

8. Use of the compound capable of targeted degradation of MYC as claimed in claim 1 in the preparation of a drug for treating leukemia.

9. Use of the compound capable of targeted degradation of MYC as claimed in claim 1 in the preparation of an anti-tumor drug in combination with doxorubicin.

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