Quinazolinone derivative as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a quinazolinone derivative as well as a preparation method and application thereof. According to the derivative disclosed by the invention, aromatic heterocyclic substitution is carried out on the C6 site of a quinazolinone mother nucleus, and diversified aromatic or heteroaromatic groups are introduced at the N3 site through linking groups (such as alkynyl, triazolyl, amido and the like) to carry out structural modification, so that a synthesis method which is mild in reaction condition, simple in step and relatively high in yield is provided. In-vivo and in-vitro antitumor activity test results show that the compound provided by the invention can inhibit BRD4 so as to resist tumors, has the advantages of high activity, good treatment effect and the like, and can be used for preparing antitumor drugs.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, and in particular relates to a quinazolinone derivative, its preparation method, and its application. Background Technology

[0002] In recent years, cancer has become one of the leading causes of death worldwide, claiming approximately 10 million lives annually from malignant tumors. Although various chemotherapy drugs are currently used in clinical treatment, their efficacy remains limited, and they are often accompanied by severe toxic side effects and drug resistance, making it difficult to effectively curb tumor progression and metastasis. Therefore, the development of novel, highly effective, and low-toxicity anti-tumor drugs has become a current research hotspot in the pharmaceutical field.

[0003] Among numerous biologically active heterocyclic compounds, quinazolinones have attracted considerable attention due to their broad pharmacological activity, particularly showing promising applications in the anti-tumor field. Existing studies have demonstrated that quinazolinone structures can inhibit tumor cell proliferation through various mechanisms, including inhibiting tyrosine kinases, interfering with the cell cycle, and inducing apoptosis. However, most existing quinazolinone compounds still suffer from insufficient activity, poor selectivity, complex synthetic routes, and harsh reaction conditions, limiting their further development and application. In particular, current research on the C6 and N3 structural modifications of the quinazolinone core is insufficient, lacking systematic structure-activity relationship analysis, making it difficult to obtain candidate compounds with both high activity and favorable pharmacokinetic properties. Furthermore, existing synthetic methods are often cumbersome, have low yields, and involve numerous side reactions, hindering large-scale preparation and subsequent drug development.

[0004] Therefore, there is an urgent need in this field to develop a class of novel quinazolinone derivatives that are structurally novel, easy to synthesize, and highly active, in order to overcome the shortcomings of existing technologies and provide new candidate molecules for the development of anti-tumor drugs. Summary of the Invention

[0005] The purpose of this invention is to provide a quinazolinone derivative, its preparation method, and its application. By modifying the structure at the C6 and N3 positions, a quinazolinone derivative with mild, simple, and efficient reaction conditions is obtained.

[0006] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0007] Synthesize key intermediates of quinazolinone core and modify the core;

[0008] A quinazolinone derivative, said quinazolinone derivative being a chemical structure having the chemical structures shown in Formula I, Formula II and Formula III or / and its pharmaceutically acceptable derivatives:

[0009]

[0010] In formulas I, II, and III, X1 and X2 are independently selected from the following groups:

[0011]

[0012] R is selected from hydrogen atom, deuterium atom, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted deuterated C1-C8 alkyl, substituted or unsubstituted C2-C8 olefinic group, substituted or unsubstituted C1-C8 alkoxy group, halogen, amino, nitro, hydroxyl, acyl, cyano, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 5-8 membered heterocyclic group containing 1-3 heteroatoms selected from N, O, S, substituted or unsubstituted 5-8 membered aryl, substituted or unsubstituted group containing 1-3 heteroatoms selected from N, O, S. 5-8 membered heteroaryl groups containing S heteroatoms; wherein the substitution is one or more substituents selected from the group consisting of: C1-C8 alkyl, halo-C1-C8 alkyl, halogen, amino, nitro, cyano, hydroxyl, C1-C8 alkoxy, halo-C1-C8 alkoxy, hydroxy-C1-C8 alkyl, C3-C8 cycloalkyl, 5-8 membered heterocyclic groups containing 1-3 N, O, or S heteroatoms, 5-8 membered aryl, and 5-8 membered heteroaryl groups containing 1-3 N, O, or S heteroatoms.

[0013] Furthermore, X and R in Formulas I, II, and III are: X1 is the common structure of series I and II, X2 is the common structure of series III, and R is selected from substituted or unsubstituted C1-C8 alkoxy groups, substituted or unsubstituted C1-C8 alkyl groups, substituted or unsubstituted C3-C8 cycloalkyl groups, substituted or unsubstituted 5-8 membered heterocyclic groups containing 1-3 heteroatoms selected from N, O, and S, substituted or unsubstituted 5-8 membered aryl groups, and substituted or unsubstituted 5-8 membered heteroaryl groups containing 1-3 heteroatoms selected from N, O, and S; wherein the substitution is one or more substituents selected from the following group: C 1- C8 alkyl, halogenated C1-C8 alkyl, halogen, amino, nitro, cyano, hydroxy, hydroxymethyl, hydroxyethyl, mercapto, carboxyl, ester, C1-C6 alkyl monosubstituted amino, C1-C6 alkyl disubstituted amino, C1-C6 alkoxy, C1-C6 alkyl carbonyloxy, C3-C6 cycloalkyl carbonyloxy, 5-8 membered heterocyclic carbonyloxy containing 1-3 heteroatoms selected from N, O, and S, C3-C6 alkyl carbonyl, C3-C6 cycloalkoxy carbonyl, 5-8 membered heterocyclic carbonyloxy containing 1-3 heteroatoms selected from N, O, and S, C1-C6 alkoxyformamide, C1-C6 alkyl mercapto.

[0014] Furthermore, pharmaceutically acceptable salts of the compounds represented by general formulas I, II, and III include: inorganic acid salts, organic acid salts, and alkyl sulfonates. Inorganic acid salts include hydrochlorides, hydrobromides, nitrates, sulfates, and phosphates; organic acid salts include formates, acetates, propionates, benzoates, maleates, fumarates, succinates, tartrates, and citrates; alkyl sulfonates include methanesulfonates, ethylsulfonates, etc.; and also include aryl sulfonates, such as benzenesulfonates and p-toluenesulfonates.

[0015] Furthermore, pharmaceutically acceptable solvates of the compounds represented by formulas I, II, and III include solvates of the compounds represented by formulas I, II, and III with water, ethanol, isopropanol, diethyl ether, and acetone.

[0016] Furthermore, the quinazolinone derivatives represented by general formulas I, II, and III have the following structural formulas:

[0017]

[0018]

[0019]

[0020] A method for preparing a quinazolinone derivative includes the following steps:

[0021] (1) Using 2-aminobenzophenone 1 as raw material, intermediate 4 is obtained by bromination, acylation and oxidation. Triphosgene is added under ice bath to carry out cyclization reaction to generate intermediate 5. Intermediate 5 is methylated by dimethyl sulfate under alkaline conditions to generate quinazolinone key intermediate A.

[0022] (2) Compound A reacts with compound B in a Suzuki reaction to give compound C;

[0023] (3) For compound I: react compound C with an alkyne reagent to introduce an alkyne group to obtain compound D, and then synthesize compound I by a click chemistry reaction of compound D with an azide compound derived from substituted aniline;

[0024] (4) For compound II: react compound C with an alkyne reagent to introduce an alkyne group to obtain compound E, hydrolyze the ester group of compound E to a carboxyl group to obtain compound F, and then synthesize compound II by condensation reaction of compound F with substituted aniline;

[0025] (5) For compound III: intermediate A is coupled with compound G via a Suzuki coupling reaction to obtain compound H. Compound H is then reacted with an alkyne reagent to introduce an alkyne group to obtain compound I. Compound I is then synthesized with an azide compound derived from substituted aniline via a click chemistry reaction to obtain compound III. The specific reaction route is as follows:

[0026] ;

[0027] .

[0028] A pharmaceutical composition comprising a quinazolinone derivative represented by formula I, formula II and formula III, a pharmaceutically acceptable salt or pharmaceutically acceptable solvate thereof, and optionally a pharmaceutically acceptable excipient.

[0029] The application of a pharmaceutical composition in the preparation of drugs for treating breast cancer, gastric cancer, and leukemia achieves a therapeutic effect by inhibiting the proliferation of breast cancer cells MCF-7, leukemia cells HL-60, or gastric cancer cells HGC-27. Attached Figure Description

[0030] Figure 1 This is a graph showing the in vivo antitumor activity test results of the compound in application I-11. Among them, (A) is a graph showing the trend of tumor volume changes during treatment; (B) is a graph showing the actual size of the dissected tumor tissue; (C) is a graph showing the changes in mouse body weight during treatment; and (D) is a graph showing the results of histopathological examination. Example

[0031] Experimental methods not specifically described in the following examples were performed under standard conditions or as recommended by the manufacturer. The structures of the compounds were determined using a Bruker-500MHz NMR spectrometer with deuterated dimethyl sulfoxide (DMSO) as the solvent and tetramethylsilane (TMS) as the internal standard. 200-300 mesh silica gel was typically used as the support for the chromatography column.

[0032] I. Examples of Compound Preparation

[0033] Example 1: Preparation of Class I Compounds

[0034]

[0035] Synthesis of compound C:

[0036] 0.54 g (1.7 mmol) of compound A and 0.4 g (1.7 mmol) of compound B were weighed and placed in a reaction flask. 12 mL of methanol was added, and the mixture was stirred to dissolve. Then, under nitrogen protection, the catalyst Pd(dppf)₂Cl₂ was added. The mixture was stirred at room temperature for 10 min, and then 3 mL of potassium carbonate solution dissolved in water was slowly added. The reaction was carried out at 110 °C for 10 h. After the reaction was completed, the reaction solution was filtered, the solvent was evaporated, and the mixture was stirred and subjected to column chromatography (DCM:MeOH = 30:1) to obtain a yellow solid product C with a yield of 71.4%.

[0037] 1 H NMR (500 MHz, Chloroform-d) δ 7.45 (dd, J = 9.4, 2.7 Hz, 1H), 7.39 (dd, J = 7.2, 1.5 Hz, 1H), 7.37 (s, 1H), 7.35-7.31 (m, 4H), 7.29 (dd, J =8.4, 2.3 Hz, 1H), 6.98 (d, J = 8.5 Hz, 1H), 6.86 (d, J = 2.2 Hz, 1H), 6.60(d, J = 9.4 Hz, 1H), 5.59 (s, 1H), 5.38 (s, 1H), 3.57 (s, 3H), 3.40 (s, 3H).

[0038] Synthesis of compound D:

[0039] 0.5 g (1.45 mmol) of compound C was weighed and placed in a reaction flask, dissolved in 5 mL of N,N-dimethylformamide, and 0.07 g (2.9 mmol) of sodium hydride was slowly added under ice bath conditions, stirring for 10 min. Then, 0.25 mL (2.9 mmol) of bromopropyne was added dropwise, and the reaction was allowed to proceed at room temperature for 1 h. After the reaction was complete, the sodium hydride was quenched with water, extracted with EA, washed with saturated brine, and dried over anhydrous sodium sulfate. The product was evaporated to dryness to give a yellow solid, product D, in 91% yield.

[0040] Synthesis of Class I compounds:

[0041] 0.2 g (1 eq) of the substituted aniline compound was added to a 50 mL round-bottom flask, along with 2 mL of hydrochloric acid and 2 mL of water. The mixture was stirred at room temperature for 20 min, then cooled to below 0°C on an ice bath. 2 mL of a solution of sodium nitrite dissolved in water (1.1 eq) was added submerged in the mixture, and the mixture was stirred for 30 min. Then, 2 mL of a solution of sodium azide dissolved in water (2 eq) was added dropwise, and the mixture was stirred for 4–8 h. After the reaction was complete, the mixture was extracted with DCM, dried over anhydrous sodium sulfate, and evaporated to dryness at low temperature to obtain the azide product.

[0042] Compound D (1 mmol) and aniline azide M1-R (2 mmol) were added to a reaction flask and dissolved in 9 mL of methanol and 3 mL of water. The mixture was stirred at room temperature until dissolved. Copper sulfate pentahydrate aqueous solution (60 mg / mL) was added, and the mixture was stirred for 10 min. Sodium ascorbate solution (32 mg / mL) was then added dropwise, and the reaction was carried out at room temperature (TLC monitoring). An appropriate amount of 200-300 mesh silica gel was added and the sample was mixed. The solvent was recovered under reduced pressure, and the corresponding compound of formula I was obtained by column chromatography on silica gel, with a yield between 50% and 70%.

[0043]

[0044] I-1: 1 H NMR (500 MHz, DMSO-d6) δ 8.31 (d, J = 2.0 Hz, 1H), 8.04 (d, J= 2.7 Hz, 1H), 7.75 (dd, J = 9.5, 2.8 Hz, 1H), 7.52 (d, J = 2.2 Hz, 1H), 7.47(dd, J = 8.5, 2.3 Hz, 1H), 7.40-7.34 (m, 4H), 7.30-7.26 (m, 1H), 7.23 (d, J =9.2 Hz, 1H), 7.18 (d, J = 3.1 Hz, 1H), 7.008 (dd, J = 9.1, 3.1 Hz, 1H), 7.05(d, J = 8.6 Hz, 1H), 6.43 (d, J = 9.4 Hz, 1H), 5.74 (s, 1H), 5.25 (d, J =15.2 Hz, 1H), 4.07 (d, J = 15.3 Hz, 1H), 3.76 (d, J = 4.1 Hz, 6H), 3.47 (s,3H). 13 C NMR (125 MHz, DMSO-d6) δ 153.36, 145.33, 138.54, 136.60, 129.00,127.88, 126.22, 125.52, 124.96, 123.13, 119.09, 115.65, 114.25, 113.94,110.92, 60.64, 56.51, 55.79, 36.99, 30.24. HRMS (ESI) m / z: calculated forC 32 H 30N6O4 [M+Na] + : 585.2226, found: 585.2215.

[0045] I-2: 1 H NMR (500 MHz, DMSO-d6) δ 8.95 (s, 1H), 8.45-8.42 (m, 2H),8.22-8.19 (m, 2H), 8.02 (d, J = 2.7 Hz, 1H), 7.72 (dd, J = 9.5, 2.8 Hz, 1H),7.49-7.45 (m, 2H), 7.40-7.35 (m, 4H), 7.30-7.27 (m, 1H), 7.06 (d, J = 8.5 Hz,1H), 6.43 (d, J = 9.4 Hz, 1H), 5.70 (s, 1H), 5.28 (d, J = 15.4 Hz, 1H), 4.08(d, J = 15.4 Hz, 1H), 3.46 (s, 3H), 3.36 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ161.02, 153.33, 146.65, 144.94, 141.54, 140.81, 138.52, 136.58, 136.35,129.82, 129.04, 127.95, 126.27, 125.55, 124.96, 124.08, 123.24, 122.54,120.52, 119.10, 116.79, 113.96, 79.19, 60.63, 36.99, 30.26. HRMS (ESI) m / z:calculated for C 30 H 25 N7O4 [M+H] + : 548.2046, found: 548.2040.

[0046] I-3: 1H NMR (500 MHz, DMSO-d6) δ 8.48 (s, 1H), 8.03 (d, J = 2.7 Hz,1H), 7.83 (d, J = 2.4 Hz, 1H), 7.79 (d, J = 8.7 Hz, 1H), 7.74 (dd, J = 9.4,2.7 Hz, 1H), 7.71 (dd, J = 8.7, 2.5 Hz, 1H), 7.50 (d, J = 2.2 Hz, 1H), 7.47(dd, J = 8.5, 2.3 Hz, 1H), 7.41-7.33 (m, 4H), 7.30-7.26 (m, 1H), 7.05 (d, J =8.5 Hz, 1H), 6.44 (d, J = 9.4 Hz, 1H), 5.75 (s, 1H), 5.25 (d, J = 15.4 Hz,1H), 4.13 (d, J = 15.3 Hz, 1H), 3.48 (s, 3H), 3.34 (s, 3H). 13 C NMR (125 MHz,DMSO-d6) δ 143.53, 141.73, 138.71, 136.77, 136.57, 135.53, 132.55, 132.10,131.56, 130.00, 129.18, 128.30, 128.05, 127.64, 126.42, 126.12, 125.14,124.36, 123.31, 119.28, 116.97, 114.12, 79.35, 60.96, 37.17, 30.42. HRMS(ESI) m / z: calculated for C 30 H 24 Cl2N6O2 [M+H] + : 571.1416, found: 571.1416.

[0047] I-4: 1H NMR (500 MHz, DMSO-d6) δ 8.61 (s, 1H), 8.02 (d, J = 2.7 Hz,1H), 7.79-7.76 (m, 2H), 7.73 (dd, J = 9.4, 2.8 Hz, 1H), 7.50 (d, J = 2.2 Hz,1H), 7.46 (dd, J = 8.5, 2.2 Hz, 1H), 7.41-7.35 (m, 4H), 7.30-7.26 (m, 1H),7.12-7.09 (m, 2H), 7.05 (d, J = 8.6 Hz, 1H), 6.43 (d, J = 9.4 Hz, 1H), 5.70(s, 1H), 5.28 (d, J = 15.3 Hz, 1H), 4.02 (d, J = 15.3 Hz, 1H), 3.82 (s, 3H),3.46 (s, 3H), 3.35 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 153.79, 144.37,141.99, 138.98, 137.04, 136.85, 130.48, 130.24, 129.48, 128.37, 126.71,125.38, 124.56, 123.66, 122.37, 122.06, 119.53, 117.25, 115.27, 114.37,79.64, 60.84, 56.01, 37.43, 30.71. HRMS (ESI) m / z: calculated for C 31 H 28 N6O3 [M+H] + : 533.2301, found:533.2308.

[0048] I-5: 1H NMR (500 MHz, DMSO-d6) δ 8.32-8.30 (m, 1H), 8.05-8.03 (m, 1H),7.78-7.72 (m, 1H), 7.60-7.56 (m, 1H), 7.54-7.49 (m, 2H), 7.48-7.45 (m, 1H),7.41- 7.34 (m, 4H), 7.31-7.26 (m, 2H), 7.15-7.10 (m, 1H), 7.06-7.02 (m, 1H),6.45-6.42 (m, 1H), 5.74 (s, 1H), 5.29-5.23 (m, 1H), 4.10-4.03 (m, 1H), 3.82(s, 3H), 3.47 (s, 3H), 3.36 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 160.86,153.20, 151.30, 1 2.74, 141.41, 138.37, 136.43, 136.26, 130.49, 129.63,128.84, 127.71, 126.06, 125.46, 125.41, 124.77, 124.02, 122.96, 120.67,118.92, 116.63, 113.76, 112.82, 79.02, 60.47, 55.92, 36.82, 30.07. HRMS (ESI)m / z: calculated for C 31 H 28 N6O3 [M+Na] + :555.2121, found:555.2117.

[0049] I-6: 1H NMR (500 MHz, DMSO-d6) δ 8.47 (d, J = 2.0 Hz, 1H), 8.03 (d, J= 2.6 Hz, 1H), 7.79 (td, J = 7.9, 1.7 Hz, 1H), 7.74 (dd, J = 9.4, 2.7 Hz,1H), 7.63-7.52 (m, 2H), 7.51 (d, J = 2.2 Hz, 1H), 7.48-7.41 (m, 2H), 7.41-7.33 (m, 4H), 7.27 (t, J = 7.0 Hz, 1H), 7.05 (dd, J = 8.6, 1.7 Hz, 1H), 6.44(d, J = 9.4 Hz, 1H), 5.75 (s, 1H), 5.25 (d, J = 15.3 Hz, 1H), 4.11 (d, J =15.3 Hz, 1H), 3.47 (s, 3H), 3.35 (d, J = 2.6 Hz, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 161.53, 155.20, 153.86, 153.21, 144.32, 142.12, 139.04, 137.10, 136.89,131.67, 130.32, 129.50, 128.39, 126.73, 126.33, 126.01, 125.65, 125.46,125.17, 124.66, 123.67, 119.59, 117.72, 117.56, 117.31, 114.44, 79.68, 61.32,37.49, 30.74. HRMS (ESI) m / z: calculated for C 30 H 26 FN6O2[M+H] + :521.2101, found:521.2094.

[0050] I-7: 1H NMR (500 MHz, DMSO-d6) δ 9.10 (s, 1H), 8.62 (s, 2H), 8.25 (s,1H), 8.01 (d, J = 2.9 Hz, 1H), 7.72 (dd, J = 9.5, 2.7 Hz, 1H), 7.49-7.45 (m,2H), 7.41-7.35 (m, 4H), 7.28 (tt, J = 6.4, 1.7 Hz, 1H), 7.06 (d, J = 8.4 Hz,1H), 6.43 (d, J = 9.4 Hz, 1H), 5.70 (s, 1H), 5.30 (d, J = 15.4 Hz, 1H), 4.09(d, J = 15.4 Hz, 1H), 3.46 (s, 3H), 3.37 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ161.18, 153.46, 145.03, 141.71, 138.67, 138.01, 136.74, 136.50, 132.10,131.83, 130.00, 129.20, 128.10, 126.46, 125.15, 124.22, 124.03, 123.41,123.07, 121.86, 120.90, 119.26, 116.94, 114.13, 79.35, 60.82, 37.14, 30.43.HRMS (ESI) m / z: calculated for C 32 H 24 F6N6O2 [M+H]+:639.1943, found:639.1937.

[0051] I-8: 1H NMR (500 MHz, DMSO-d6) δ 8.81 (s, 1H), 8.02 (d, J = 2.9 Hz,1H), 7.83 (dt, J = 10.0, 2.3 Hz, 1H), 7.79 (ddd, J = 8.1, 2.1, 0.9 Hz, 1H),7.73 (dd, J = 9.5, 2.7 Hz, 1H), 7.63 (td, J = 8.3, 6.3 Hz, 1H), 7.49 (d, J =2.2 Hz, 1H), 7.46 (dd, J = 8.5, 2.3 Hz, 1H), 7.41-7.35 (m, 4H), 7.33 (ddd, J= 8.6, 2.5, 0.9 Hz, 1H), 7.30-7.26 (m, 1H), 7.05 (d, J = 8.5 Hz, 1H), 6.45-6.41 (m, 1H), 5.71-5.67 (m, 1H), 5.29 (d, J = 15.4 Hz, 1H), 4.04 (d, J = 15.3Hz, 1H), 3.46 (s, 3H), 3.36 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 161.37,153.67, 144.74, 141.86, 138.87, 136.93, 136.70, 132.20, 130.15, 129.39,128.29, 126.61, 125.30, 124.42, 123.58, 122.57, 119.43, 117.14, 116.21,115.59, 114.29, 107.88, 107.6, 79.53, 60.82, 37.33, 30.61. HRMS (ESI) m / zCalculated for C 30 H 25 FN6O2 [M+H] + :521.2101, found:521.2095.

[0052] I-9: 1H NMR (500 MHz, DMSO-d6) δ 8.47 (s, 1H), 8.21 (s, 1H), 8.07 (s,1H), 8.03 (d, J = 2.7 Hz, 1H), 7.74 (dd, J = 9.4, 2.8 Hz, 1H), 7.50 (d, J =2.2 Hz, 1H), 7.47 (dd, J = 8.5, 2.2 Hz, 1H), 7.40-7.33 (m, 4H), 7.28 (tt, J =6.3, 1.7 Hz, 1H), 7.05 (d, J = 8.5 Hz, 1H), 6.44 (d, J = 9.4 Hz, 1H), 5.74(s, 1H), 5.24 (d, J = 15.4 Hz, 1H), 4.12 (d, J = 15.4 Hz, 1H), 3.48 (s, 3H),3.34 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 161.54, 153.84, 143.98, 142.07,139.05, 137.11, 136.90, 134.62, 134.18, 132.19, 131.38, 130.35, 130.04,129.53, 128.81, 128.41, 126.76, 126.52, 125.49, 124.69, 123.65, 119.62,117.30, 114.47, 79.69, 61.36, 37.52, 30.76. HRMS (ESI) m / z Calculated forC 30 H 23 Cl3N6O2 [M+H] + :605.1026, found:605.1027.

[0053] I-10: 1H NMR (500 MHz, DMSO-d6) δ 8.97 (d, J = 2.5 Hz, 1H), 8.78 (s,1H), 8.72 (dd, J = 8.8, 2.5 Hz, 1H), 8.03 (d, J = 2.7 Hz, 1H), 7.74 (dd, J =9.4, 2.8 Hz, 1H), 7.49-7.46 (m, 2H), 7.40-7.36 (m, 4H), 7.30-7.26 (m, 1H),7.09-7.04 (m, 1H), 6.44 (d, J = 9.4 Hz, 1H), 5.74 (s, 1H), 5.24 (d, J = 15.5Hz, 1H), 4.14 (d, J = 15.4 Hz, 1H), 3.47 (s, 3H), 3.41(s,1H),3.36(s,3H). 13 CNMR (125 MHz, DMSO-d6) δ 161.05, 153.34, 147.34, 144.63, 141.55, 139.89,138.56, 136.63, 136.37, 132.79, 129.90, 129.05, 128.61, 128.12, 127.98,126.28, 125.05, 124.13, 123.20, 121.32, 119.14, 116.84, 114.02, 105.00,61.00, 37.02, 30.27. HRMS (ESI) m / z Calculated for C 30 H 24 N8O6 [M+H] + :593.1897,found:593.1896.

[0054] I-11: 1H NMR (500 MHz, DMSO-d6) δ 8.78 (s, 1H), 8.03 (d, J = 2.7 Hz,1H), 7.74 (dd, J = 9.5, 2.7 Hz, 1H), 7.52 (d, J = 2.2 Hz, 1H), 7.46 (dd, J =8.5, 2.2 Hz, 1H), 7.42-7.36 (m, 4H), 7.29 (ddd, J = 8.5, 5.9, 2.2 Hz, 1H),7.18 (s, 2H), 7.05 (d, J = 8.6 Hz, 1H), 6.43 (d, J = 9.4 Hz, 1H), 5.68 (s,1H), 5.31 (d, J = 15.4 Hz, 1H), 4.03 (d, J = 15.2 Hz, 1H), 3.86 (s, 6H), 3.70(s, 3H), 3.46 (s, 3H), 3.36 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 161.04,153.48, 153.36, 144.04, 141.48, 138.53, 137.26, 136.60, 136.38, 132.50,129.83, 129.07, 127.96, 126.25, 124.95, 124.10, 123.20, 122.31, 119.09,116.77, 113.95, 97.94, 79.19, 60.36, 60.21, 56.29, 37.00, 30.28. HRMS (ESI)m / z Calculated for C 33 H 32 N6O5 [M+H] + :593.2512, found:593.2513.

[0055] I-12: 1H NMR (500 MHz, DMSO-d6) δ 8.76 (s, 1H), 8.02 (d, J = 2.7 Hz,1H), 7.95-7.92 (m, 2H), 7.72-7.70 (m, 2H), 7.49 (d, J = 2.2 Hz, 1H), 7.46(dd, J = 8.5, 2.2 Hz, 1H), 7.40-7.34 (m, 4H), 7.28 (ddt, J = 8.5, 6.3, 1.9Hz, 1H), 7.05 (d, J = 8.5 Hz, 1H), 6.43 (d, J = 9.4 Hz, 1H), 5.69 (s, 1H),5.28 (d, J = 15.3 Hz, 1H), 4.03 (d, J = 15.3 Hz, 1H), 3.46 (s, 3H), 3.41 (s,1H), 3.35 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 161.03, 153.32, 144.41,141.54, 139.88, 138.55, 136.58, 136.37, 136.24, 129.80, 129.03, 127.93,126.26, 124.94, 124.09, 123.23, 121.94, 121.85, 119.16, 119.09, 116.80,113.93, 104.99, 94.26, 60.50, 36.99, 30.26. HRMS (ESI) m / z Calculated forC 30 H 25 IN6O2 [M+H] + :629.1162, found:629.1163.

[0056] I-13: 1H NMR (500 MHz, DMSO-d6) δ 8.77 (s, 1H), 8.02 (d, J = 2.7 Hz,1H), 7.88-7.85 (m, 2H), 7.80-7.77 (m, 2H), 7.73 (dd, J = 9.5, 2.7 Hz, 1H),7.50 (d, J = 2.2 Hz, 1H), 7.46 (dd, J = 8.5, 2.2 Hz, 1H), 7.41-7.36 (m, 4H),7.30-7.26 (m, 1H), 7.05 (d, J = 8.6 Hz, 1H), 6.43 (d, J = 9.4 Hz, 1H), 5.70(s, 1H), 5.28 (d, J = 15.4 Hz, 1H), 4.04 (d, J = 15.3 Hz, 1H), 3.46 (s, 3H),3.36 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 144.42, 142.02, 141.54, 138.52,136.59, 136.37, 135.78, 132.75, 129.80, 129.04, 127.94, 126.27, 123.23,122.07, 121.89, 121.25, 119.09, 116.80, 114.01-113.88 (m), 104.99, 60.51,37.00, 30.26. HRMS (ESI) m / z Calculated for C 30 H 25 BrN6O2 [M+H] + :581.1301,found:581.1309.

[0057] I-14: 1H NMR (500 MHz, DMSO-d6) δ 8.46 (s, 1H), 8.04 (d, J = 2.7 Hz,1H), 7.88 (d, J = 2.4 Hz, 1H), 7.84 (d, J = 8.7 Hz, 1H), 7.76 (d, J = 2.4 Hz,1H), 7.74 (d, J = 4.1 Hz, 1H), 7.50 (d, J = 2.2 Hz, 1H), 7.47 (dd, J = 8.5,2.2 Hz, 1H), 7.40-7.36 (m, 4H), 7.30-7.26 (m, 1H), 7.05 (d, J = 8.5 Hz, 1H),6.45 (d, J = 9.4 Hz, 1H), 5.72 (s, 1H), 5.26 (d, J = 15.3 Hz, 1H), 4.11 (d, J= 15.4 Hz, 1H), 3.48 (s, 3H), 3.35 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ161.05, 153.35, 143.21, 139.88, 138.56, 137.29, 136.61, 136.42, 135.15,134.62, 131.21, 129.84, 129.03, 127.89, 126.28, 126.00, 124.20, 123.16,120.94, 119.16, 118.26, 116.81, 113.97, 105.00, 79.19, 60.65, 37.03, 36.74,30.28. HRMS (ESI) m / z Calculated for C 30 H 24 Br2N6O2 [M+Na] + :681.0225, found:681.0221.

[0058] I-15: 1H NMR (500 MHz, DMSO-d6) δ 8.77 (s, 1H), 8.03 (d, J = 2.7 Hz,1H), 7.94-7.92 (m, 2H), 7.73 (dd, J = 9.4, 2.7 Hz, 1H), 7.67-7.64 (m, 2H),7.50 (d, J = 2.3 Hz, 1H), 7.46 (dd, J = 8.5, 2.2 Hz, 1H), 7.41-7.36 (m, 4H),7.30-7.26 (m, 1H), 7.05 (d, J = 8.6 Hz, 1H), 6.43 (d, J = 9.4 Hz, 1H), 5.69(s, 1H), 5.29 (d, J = 15.4 Hz, 1H), 4.04 (d, J = 15.3 Hz, 1H), 3.46 (s, 3H),3.36 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 161.03, 153.33, 144.39, 141.54,138.52, 136.58, 136.37, 135.38, 132.86, 129.83, 129.03, 127.93, 126.27,124.94, 124.09, 123.23, 122.12, 121.65, 119.09, 116.80, 113.93, 79.19, 60.50,53.24, 36.99, 30.26. HRMS (ESI) m / z Calculated for C 30 H 25 ClN6O2 [M+H] + :537.1806, found:537.1809.

[0059] I-16: 1H NMR (500 MHz, DMSO-d6) δ 8.46 (d, J = 1.8 Hz, 1H), 8.04 (d, J= 2.7 Hz, 1H), 7.85 (td, J = 8.8, 5.9 Hz, 1H), 7.75 (dd, J = 9.4, 2.7 Hz,1H), 7.67 (ddd, J = 11.2, 8.9, 2.8 Hz, 1H), 7.51 (d, J = 2.2 Hz, 1H), 7.47(dd, J = 8.5, 2.2 Hz, 1H), 7.40-7.31 (m, 5H), 7.28 (t, J = 6.8 Hz, 1H), 7.05(d, J = 8.5 Hz, 1H), 6.44 (d, J = 9.4 Hz, 1H), 5.74 (s, 1H), 5.24 (d, J =15.4 Hz, 1H), 4.10 (d, J = 15.3 Hz, 1H), 3.47 (s, 3H), 3.34 (s, 3H). 13 C NMR(125 MHz, DMSO-d6) δ 161.04, 153.35, 143.82, 141.61, 138.55, 136.61, 136.38,129.82, 129.01, 127.90, 127.61, 127.53, 126.23, 125.31, 124.97, 124.16,123.18, 119.11, 116.81, 113.96, 112.75, 112.60, 79.19, 60.85, 37.00, 30.24.HRMS (ESI) m / z Calculated for C 30 H 24 F2N6O2 [M+H] + :539.2007, found:539.2007.

[0060] I-17: 1H NMR (500 MHz, DMSO-d6) δ 8.81 (s, 1H), 8.05-7.99 (m, 3H),7.73 (dd, J = 9.5, 2.7 Hz, 1H), 7.50-7.45 (m, 2H), 7.37 (d, J = 6.8 Hz, 4H),7.31-7.26 (m, 1H), 7.06 (d, J = 8.4 Hz, 1H), 6.43 (d, J = 9.4 Hz, 1H), 5.68(s, 1H), 5.27 (d, J = 15.4 Hz, 1H), 4.04 (d, J = 15.4 Hz, 1H), 3.46 (s, 3H),3.36 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 161.03, 153.31, 144.77, 141.51,138.52, 136.59, 136.33, 129.83, 129.05, 127.96, 126.27, 124.97, 124.05,123.24, 122.47, 119.11, 116.79, 113.97, 105.66, 105.45, 60.64, 37.00, 30.26.HRMS (ESI) m / z Calculated for C 30 H 23 F3N6O2 [M+H] + :557.1913, found:557.1913.

[0061] I-18: 1H NMR (500 MHz, DMSO-d6) δ 8.73 (s, 1H), 8.03 (d, J = 2.7 Hz,1H), 7.95-7.90 (m, 2H), 7.73 (dd, J = 9.5, 2.8 Hz, 1H), 7.50 (d, J = 2.2 Hz,1H), 7.48-7.42 (m, 3H), 7.41-7.35 (m, 4H), 7.30-7.26 (m, 1H), 7.05 (d, J =8.5 Hz, 1H), 6.43 (d, J = 9.4 Hz, 1H), 5.70 (s, 1H), 5.29 (d, J = 15.3 Hz,1H), 4.03 (d, J = 15.3 Hz, 1H), 3.46 (s, 3H), 3.36 (s, 3H). 13 C NMR (125 MHz,DMSO-d6) δ 161.03, 160.61, 153.34, 144.23, 141.54, 138.53, 136.59, 136.38,133.17, 129.80, 129.04, 127.93, 126.27, 124.95, 124.10, 123.23, 122.36,122.29, 119.09, 116.79, 116.61, 113.94, 60.46, 36.99, 30.26. HRMS (ESI) m / zCalculated for C 30 H 25 FN6O2 [M+Na] + :543.1921, found:543.1911.

[0062] I-19: 1H NMR (500 MHz, DMSO-d6) δ 8.50 (s, 1H), 8.05 (d, J = 2.7 Hz,1H), 7.76 (dd, J = 9.4, 2.7 Hz, 1H), 7.74-7.68 (m, 1H), 7.53 (d, J = 2.2 Hz,1H), 7.49-7.42 (m, 3H), 7.37 (dd, J = 13.9, 6.6 Hz, 4H), 7.31-7.25 (m, 1H),7.05 (d, J = 8.5 Hz, 1H), 6.44 (d, J = 9.4 Hz, 1H), 5.78 (s, 1H), 5.23 (d, J= 15.4 Hz, 1H), 4.13 (d, J = 15.4 Hz, 1H), 3.48 (s, 3H), 3.34 (s, 3H). 13 C NMR(125 MHz, DMSO-d6) δ 161.05, 153.35, 143.69, 141.61, 138.57, 136.64, 136.37,132.64, 129.87, 129.02, 127.92, 126.63, 126.20, 125.01, 124.17, 123.15,119.11, 116.82, 113.98, 112.95, 112.77, 79.19, 61.19, 37.01, 30.24. HRMS(ESI) m / z Calculated for C 30 H 25 FN6O2 [M+H] + :539.2007, found:539.2003

[0063] I-20: 1H NMR (500 MHz, DMSO-d6) δ 8.86 (s, 1H), 8.03 (d, J = 2.7 Hz,1H), 7.80-7.76 (m, 2H), 7.73 (dd, J = 9.5, 2.7 Hz, 1H), 7.50-7.45 (m, 2H),7.43 (dt, J = 9.3, 2.3 Hz, 1H), 7.41-7.35 (m, 5H), 7.31-7.26 (m, 1H), 7.06(d, J = 8.5 Hz, 1H), 6.43 (d, J = 9.4 Hz, 1H), 5.68 (s, 1H), 5.28 (d, J =15.4 Hz, 1H), 4.03 (d, J = 15.3 Hz, 1H), 3.46 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 161.03, 153.32, 144.64, 141.50, 139.88, 139.84, 138.53, 136.59, 136.35,129.83, 129.05, 127.96, 126.28, 124.97, 124.06, 123.25, 122.40, 119.16,119.10, 116.80, 113.96, 104.99, 103.91, 103.67, 60.56, 36.99, 30.27. HRMS(ESI) m / z Calculated for C 30 H 25 FN6O2 [M+H] + :539.2007, found:539.2007.

[0064] Example 2: Preparation of Class II Compounds

[0065]

[0066] Synthesis of compound E:

[0067] 0.5 g (1.45 mmol) of compound C was dissolved in N,N-dimethylformamide. 0.07 g (2.9 mmol) of sodium hydride was slowly added under ice bath conditions, and the mixture was stirred for 10 min. Then, 0.24 mL (2.16 mmol) of bromopropyne was added dropwise, and the reaction was allowed to proceed at room temperature for 4 h. After the reaction was complete, water was added to quench the reaction mixture. The reaction solution was extracted with ethyl acetate, the organic phase was washed with saturated brine, and dried over anhydrous sodium sulfate. The product was evaporated to dryness to give a yellow solid, product E, in 88.3% yield.

[0068] Synthesis of compound F:

[0069] Compound E (0.6 g, 1.39 mmol) and 4 mL of methanol were added to a reaction flask and stirred until dissolved. Then, 0.17 g (6.95 mmol) of lithium hydroxide was added at room temperature, and the reaction was allowed to proceed for approximately 2 h at room temperature (monitored by TLC). After the reaction was complete, the reaction solution was concentrated, and the pH was adjusted to 5 with 1 M hydrochloric acid. A white solid precipitated from the system. The product was filtered, washed twice with distilled water, and dried to obtain a white solid powder F, with a yield of 89%.

[0070] Synthesis of Class II compounds:

[0071] Compound F (0.2 mmol), EDC·HCl (0.4 mmol), and DMAP (0.3 mmol) were added to a reaction flask and dissolved in 15 mL of dichloromethane. The mixture was stirred at room temperature for 10 min, and then M2-R (0.4 mmol) was added. The reaction was continued at room temperature (detected by TLC). A suitable amount of 200-300 mesh silica gel was added as a sample, the solvent was recovered under reduced pressure, and the corresponding class II compound was obtained by column chromatography on silica gel, with yields between 50% and 80%.

[0072]

[0073] II-1: 1H NMR (500 MHz, DMSO-d6) δ 8.05 (d, J = 2.7 Hz, 1H), 7.75 (dd,J = 9.5, 2.7 Hz, 1H), 7.50 (dd, J = 8.5, 2.2 Hz, 1H), 7.44 (d, J = 2.1 Hz,1H), 7.37-7.32 (m, 4H), 7.28 (td, J = 6.0, 2.6 Hz, 1H), 7.08 (d, J = 8.5 Hz,1H), 6.46 (d, J = 9.4 Hz, 1H), 5.68 (s, 1H), 4.40 (d, J = 17.3 Hz, 1H), 3.74(d, J = 17.3 Hz, 1H), 3.61 (s, 3H), 3.49 (s, 3H), 3.37 (s, 10H), 3.31 (s,3H). 13 C NMR (125 MHz, DMSO-d6) δ 170.11, 161.51, 153.67, 142.10, 139.04,137.13, 136.84, 130.43, 129.40, 128.42, 126.80, 125.57, 124.51, 123.65,119.59, 117.31, 114.50, 62.88, 52.26, 48.51, 37.48, 30.57. HRMS (ESI) m / zCalculated for C 32 H 32 N4O6 [M+Na] + :591.2220, found:591.1926.

[0074] II-2: 1H NMR (500 MHz, DMSO-d6) δ 9.78 (s, 1H), 8.10 (s, 1H), 8.05 (d,J = 2.7 Hz, 1H), 7.91 (s, 1H), 7.75 (dd, J = 9.4, 2.8 Hz, 1H), 7.50 (dd, J =8.4, 2.0 Hz, 1H), 7.46 (d, J = 2.2 Hz, 1H), 7.39-7.33 (m, 4H), 7.27 (tt, J =5.7, 3.0 Hz, 1H), 7.09 (d, J = 8.6 Hz, 1H), 6.46 (d, J = 9.4 Hz, 1H), 5.69(s, 1H), 4.65 (d, J = 16.9 Hz, 1H), 3.81 (d, J = 16.9 Hz, 1H), 3.48 (s, 3H),3.33 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 168.22, 166.04, 163.81, 156.10,155.84, 153.66, 153.47, 145.65, 141.98, 138.76, 136.83, 136.59, 134.75,130.71, 129.17, 128.14, 126.54, 124.30, 123.42, 122.42, 121.13, 119.33,37.20, 30.40. HRMS (ESI) m / z Calculated for C 29 H 23 Cl3N4O3 [M+Na] + :603.0733,found:603.0725.

[0075] II-3: 1H NMR (500 MHz, DMSO-d6) δ 9.52 (s, 1H), 8.05 (d, J = 2.7 Hz,1H), 7.76 (dd, J = 9.5, 2.7 Hz, 1H), 7.67 (d, J = 7.7 Hz, 1H), 7.64 (dd, J =8.0, 1.2 Hz, 1H), 7.52-7.47 (m, 2H), 7.39-7.34 (m, 5H), 7.28 (tt, J = 5.6,2.7 Hz, 1H), 7.14-7.06 (m, 2H), 6.46 (d, J = 9.4 Hz, 1H), 5.71 (s, 1H), 4.66(d, J = 16.8 Hz, 1H), 3.70 (d, J = 16.8 Hz, 1H), 3.48 (s, 3H), 3.34 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 167.77, 161.51, 153.97, 142.11, 139.03, 137.08,136.94, 136.26, 133.08, 130.36, 129.48, 128.53, 128.45, 127.33, 126.79,126.53, 125.49, 124.62, 123.70, 119.60, 117.33, 114.49, 62.77, 37.49, 30.71.HRMS (ESI) m / z Calculated for C 29 H 25 BrN4O3 [M+Na] + :579.1008, found:579.1000.

[0076] II-4: 1H NMR (500 MHz, DMSO-d6) δ 9.89 (s, 1H), 8.05 (d, J = 2.7 Hz,1H), 7.76 (dd, J = 9.5, 2.7 Hz, 1H), 7.51-7.46 (m, 4H), 7.39-7.35 (m, 4H),7.29 (dt, J = 8.7, 4.2 Hz, 1H), 7.08 (d, J = 8.4 Hz, 1H), 6.89-6.85 (m, 2H),6.46 (d, J = 9.4 Hz, 1H), 5.67 (s, 1H), 4.59 (d, J = 16.5 Hz, 1H), 3.71 (s,3H), 3.55 (d, J = 16.5 Hz, 1H), 3.49 (s, 3H), 3.32 (s, 3H). 13 C NMR (125 MHz,DMSO-d6) δ 166.82, 161.54, 155.64, 153.88, 142.42, 139.04, 137.06, 136.99,132.47, 130.24, 129.47, 128.39, 126.73, 125.44, 124.57, 123.69, 121.03,119.59, 117.41, 114.30, 62.79, 55.59, 49.74, 37.49, 30.61. HRMS (ESI) m / zCalculated for C 30 H 28 N4O4 [M+Na] + :531.2008, found:531.1998.

[0077] II-5: 1H NMR (500 MHz, DMSO-d6) δ 8.05 (d, J = 2.7 Hz, 1H), 7.75 (dd,J = 9.4, 2.8 Hz, 1H), 7.50 (dd, J = 8.5, 2.3 Hz, 1H), 7.43 (d, J = 2.2 Hz,1H), 7.34 (d, J = 6.5 Hz, 4H), 7.27 (ddt, J = 8.9, 6.1, 3.1 Hz, 1H), 7.08 (d,J = 8.6 Hz, 1H), 6.46 (d, J = 9.4 Hz, 1H), 5.67 (s, 1H), 4.39 (d, J = 17.2Hz, 1H), 3.74 (d, J = 17.2 Hz, 1H), 3.60 (s, 3H), 3.48 (s, 3H), 3.30 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 170.11, 161.51, 153.67, 142.10, 139.05, 137.14,136.84, 130.43, 129.40, 128.42, 126.80, 125.57, 124.51, 123.65, 119.60,117.30, 114.51, 62.87, 52.26, 48.51, 37.48, 30.57. HRMS (ESI) m / z Calculatedfor C 29 H 24 Cl2N4O3 [M+Na] + :519.1123, found:519.1117.

[0078] II-6: 1H NMR (500 MHz, DMSO-d6) δ 9.61 (s, 1H), 8.05 (d, J = 2.7 Hz,1H), 7.91 (d, J = 2.4 Hz, 1H), 7.76 (dd, J = 9.4, 2.8 Hz, 1H), 7.60 (d, J =8.6 Hz, 1H), 7.51 (dd, J = 8.5, 2.2 Hz, 1H), 7.47 (d, J = 2.2 Hz, 1H), 7.39-7.33 (m, 4H), 7.30 (ddd, J = 11.1, 7.3, 2.8 Hz, 2H), 7.10 (d, J = 8.5 Hz,1H), 6.46 (d, J = 9.4 Hz, 1H), 5.72 (s, 1H), 4.63 (d, J = 16.8 Hz, 1H), 3.78(d, J = 16.8 Hz, 1H), 3.49 (s, 3H), 3.34 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ168.20, 161.53, 153.97, 142.18, 139.04, 137.77, 137.10, 136.89, 134.70,130.40, 129.65, 129.46, 128.44, 126.82, 125.53, 124.62, 123.71, 120.77,119.60, 118.35, 117.34, 114.54, 79.63, 62.83, 37.49, 30.71. HRMS (ESI) m / zCalculated for C 29 H 24 Br2N4O3 [M+Na] + :657.0113, found:657.0107.

[0079] II-7: 1H NMR (500 MHz, DMSO-d6) δ 10.16 (s, 1H), 8.05 (d, J = 2.7 Hz,1H), 7.76 (dd, J = 9.5, 2.7 Hz, 1H), 7.62-7.57 (m, 2H), 7.53-7.46 (m, 2H),7.40-7.33 (m, 6H), 7.29 (h, J = 4.1 Hz, 1H), 7.08 (d, J = 8.5 Hz, 1H), 6.46(d, J = 9.4 Hz, 1H), 5.68 (s, 1H), 4.58 (d, J = 16.5 Hz, 1H), 3.62 (d, J =16.5 Hz, 1H), 3.49 (s, 3H), 3.32 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 167.55,161.54, 153.88, 142.39, 139.04, 138.27, 137.07, 136.95, 130.29, 129.47,129.13, 128.41, 127.26, 126.74, 125.48, 124.54, 123.69, 121.04, 119.59,117.40, 114.42, 62.88, 50.03, 37.49, 30.60. HRMS (ESI) m / z Calculated forC 29 H 25 ClN4O3 [M+Na] + :535.1513, found:535.1505.

[0080] II-8: 1H NMR (500 MHz, DMSO-d6) δ 8.04 (d, J = 2.7 Hz, 1H), 7.75 (dd,J = 9.4, 2.7 Hz, 1H), 7.50 (dd, J = 8.5, 2.1 Hz, 1H), 7.43 (d, J = 2.1 Hz,1H), 7.35 (dt, J = 14.0, 6.3 Hz, 5H), 7.27 (ddd, J = 8.5, 5.4, 2.1 Hz, 1H),7.08 (d, J = 8.5 Hz, 1H), 6.46 (d, J = 9.4 Hz, 1H), 5.68 (s, 1H), 4.39 (d, J= 17.3 Hz, 1H), 3.74 (d, J = 17.3 Hz, 1H), 3.61 (s, 3H), 3.49 (s, 3H), 3.31(s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 170.11, 161.54, 153.67, 142.09, 139.07,137.13, 136.84, 130.43, 129.40, 128.43, 126.79, 125.59, 124.52, 123.65,119.59, 117.35, 114.51, 62.87, 52.27, 48.51, 37.49, 30.57. HRMS (ESI) m / zCalculated for C 29 H 25 ClN4O3 [M+Na] + :535.1513, found:535.1506.

[0081] II-9: 1H NMR (500 MHz, DMSO-d6) δ 9.22 (s, 1H), 8.06 (d, J = 2.7 Hz,1H), 7.97 (d, J = 7.9 Hz, 1H), 7.76 (dd, J = 9.5, 2.7 Hz, 1H), 7.54-7.48 (m,2H), 7.38-7.33 (m, 4H), 7.28 (tt, J = 5.7, 3.0 Hz, 1H), 7.10 (d, J = 8.4 Hz,1H), 7.05 (t, J = 7.8 Hz, 1H), 7.01 (d, J = 8.1 Hz, 1H), 6.89 (t, J = 7.6 Hz,1H), 6.46 (d, J = 9.4 Hz, 1H), 5.69 (s, 1H), 4.66 (d, J = 16.8 Hz, 1H), 3.77-3.69 (m, 4H), 3.49 (s, 3H), 3.33 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 167.42,161.54, 153.98, 142.22, 139.04, 137.08, 136.98, 130.35, 129.43, 128.38,127.46, 126.72, 125.49, 124.70, 123.66, 121.42, 120.73, 119.61, 117.40,114.46, 111.54, 79.62, 62.80, 56.08, 37.49, 30.67. HRMS (ESI) m / z Calculatedfor C 30 H 28 N4O4 [M+Na] + :531.2008, found:531.1998.

[0082] II-10: 1H NMR (500 MHz, DMSO-d6) δ 8.05 (t, J = 3.0 Hz, 1H), 7.76 (dt,J = 9.4, 3.3 Hz, 1H), 7.50 (dt, J = 8.5, 2.7 Hz, 1H), 7.45 (dd, J = 18.8, 2.2Hz, 1H), 7.38 – 7.32 (m, 4H), 7.28 (dt, J = 9.7, 3.2 Hz, 1H), 7.08 (dd, J =8.6, 2.9 Hz, 1H), 6.46 (d, J = 9.4 Hz, 1H), 5.68 (d, J = 3.8 Hz, 1H), 4.39(d, J = 17.2 Hz, 1H), 3.74 (d, J = 17.2 Hz, 1H), 3.60 (s, 2H), 3.49 (s, 3H),3.31 (d, J = 7.4 Hz, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 170.11, 161.55, 153.67,142.09, 139.07, 137.13, 136.84, 130.42, 129.47, 129.40, 128.43, 126.79,126.75, 125.59, 124.52, 123.65, 119.59, 117.36, 114.51, 112.42, 107.79,107.34, 102.97, 99.35, 62.87, 52.27, 48.51, 37.49, 30.57. HRMS (ESI) m / zCalculated for C 29 H 24 F2N4O3 [M+Na] + :537.1714, found:537.1707.

[0083] II-11: 1H NMR (500 MHz, DMSO-d6) δ 8.05 (s, 1H), 7.75 (dd, J = 9.4,2.3 Hz, 1H), 7.51 – 7.43 (m, 2H), 7.41 – 7.31 (m, 5H), 7.31 – 7.25 (m, 1H),7.16 (t, J = 8.2 Hz, 1H), 7.08 (dd, J = 8.5, 4.9 Hz, 1H), 6.46 (dd, J = 9.4,3.8 Hz, 1H), 5.67 (d, J = 9.9 Hz, 1H), 4.74 (d, J = 16.8 Hz, 1H), 3.61 (s,2H), 3.48 (d, J = 2.8 Hz, 3H), 3.32 (d, J = 11.3 Hz, 3H). 13 C NMR (125 MHz,DMSO-d6) δ 170.11, 161.55, 153.86, 153.68, 139.07, 137.13, 137.07, 136.92,136.84, 130.42, 130.28, 129.54, 129.40, 128.49, 128.43, 126.79, 126.74,125.59, 125.47, 119.59, 117.38, 114.52, 114.43, 62.87, 62.71, 52.27, 37.49,30.66, 30.57. HRMS (ESI) m / z Calculated for C 29 H 24 F2N4O3 [M+Na] + :537.1714,found:537.1705.

[0084] II-12: 1H NMR (500 MHz, DMSO-d6) δ 10.13 (s, 1H), 8.06 (d, J = 2.8 Hz,1H), 7.76 (dd, J = 9.5, 2.7 Hz, 1H), 7.65-7.62 (m, 2H), 7.52-7.46 (m, 2H),7.41 (d, J = 8.8 Hz, 2H), 7.37 (d, J = 4.3 Hz, 4H), 7.29 (h, J = 4.0 Hz, 1H),7.08 (d, J = 8.5 Hz, 1H), 6.46 (d, J = 9.4 Hz, 1H), 5.76 (s, 1H), 5.68 (s,1H), 4.57 (d, J = 16.5 Hz, 1H), 3.62 (d, J = 16.5 Hz, 1H), 3.49 (s, 3H), 3.32(s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 167.57, 153.87, 142.39, 139.15, 139.04,137.85, 137.08, 129.46, 126.74, 124.54, 123.69, 121.69, 119.59, 117.37,114.41, 87.19, 62.88, 55.38, 50.08, 39.68, 37.49, 30.60. HRMS (ESI) m / zCalculated for C 29 H 25 IN4O3 [M+Na] + :627.0869, found:627.0862.

[0085] II-13: 1H NMR (500 MHz, DMSO-d6) δ 10.40 (s, 1H), 8.05 (d, J = 2.7 Hz,1H), 7.76 (dd, J = 9.4, 2.7 Hz, 1H), 7.50 (dd, J = 8.5, 2.2 Hz, 1H), 7.49-7.43 (m, 3H), 7.36 (d, J = 3.7 Hz, 4H), 7.29 (ddd, J = 8.7, 5.8, 3.2 Hz, 1H),7.09 (d, J = 8.5 Hz, 1H), 6.46 (d, J = 9.4 Hz, 1H), 5.68 (s, 1H), 4.53 (d, J= 16.5 Hz, 1H), 3.66 (d, J = 16.5 Hz, 1H), 3.48 (s, 3H), 3.32 (s, 3H). 13 C NMR(125 MHz, DMSO-d6) δ 170.11, 161.55, 153.67, 139.07, 137.13, 129.47, 129.40,128.43, 126.79, 126.75, 125.59, 124.52, 119.59, 117.36, 114.51, 62.87, 52.27,48.51, 39.67, 37.49, 30.57. HRMS (ESI) m / z Calculated for C 29 H 23 F3N4O3 [M+Na] + :555.1620, found:555.1611.

[0086] II-14: 1H NMR (500 MHz, DMSO-d6) δ 9.86 (s, 1H), 8.05 (d, J = 2.7 Hz,1H), 7.83 (td, J = 9.0, 6.2 Hz, 1H), 7.75 (dd, J = 9.5, 2.7 Hz, 1H), 7.52 –7.48 (m, 1H), 7.45 (dd, J = 15.3, 2.2 Hz, 1H), 7.40 – 7.33 (m, 4H), 7.33 –7.26 (m, 2H), 7.10 – 7.03 (m, 2H), 6.46 (d, J = 9.4 Hz, 1H), 5.67 (s, 1H),4.67 (d, J = 16.8 Hz, 1H), 3.67 (d, J = 16.8 Hz, 1H), 3.48 (s, 3H), 3.32 (s,3H). 13 C NMR (125 MHz, DMSO-d6) δ 167.92, 161.52, 153.87, 139.03, 137.07,136.95, 130.29, 129.46, 128.42, 126.77, 125.46, 124.56, 123.68, 119.59,117.37, 114.41, 62.83, 49.66, 39.70, 37.48, 30.62. HRMS (ESI) m / z Calculatedfor C 29 H 23 F2N4O3 [M+Na] + :537.1714, found:537.1707.

[0087] II-15: 1H NMR (500 MHz, DMSO-d6) δ 8.05 (d, J = 2.6 Hz, 1H), 7.75 (dt,J = 9.4, 2.7 Hz, 1H), 7.50 (ddd, J = 8.4, 5.7, 2.2 Hz, 1H), 7.45 (dd, J =17.5, 2.2 Hz, 1H), 7.39-7.31 (m, 4H), 7.31-7.25 (m, 1H), 7.09 (t, J = 8.5 Hz, 1H), 6.46 (d, J = 9.4 Hz, 1H), 5.69 (d, J = 19.5 Hz, 1H), 4.49 (dd, J = 95.3,16.9 Hz, 1H), 3.73 (dd, J = 16.9, 9.9 Hz, 1H), 3.60 (s, 2H), 3.48 (s, 3H), 3.32 (d, J = 11.9 Hz, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 161.53, 142.09,139.06, 137.14, 129.49, 129.40, 128.43, 126.80, 124.51, 123.65, 119.59,114.51, 62.87, 52.27, 48.51, 39.68, 39.50, 37.49, 30.57. HRMS (ESI) m / zCalculated for C 31 H 24 F6N4O3 [M+Na] + :637.1650, found:637.1646.

[0088] Example 3: Preparation of Class III Compounds

[0089]

[0090] Synthesis of compound H:

[0091] 0.3 g (0.95 mmol) of compound A and 0.32 g (1.45 mmol) of compound G were weighed and placed in a reaction flask. 8 mL of methanol was added, and the mixture was stirred to dissolve. Then, under nitrogen protection, the catalyst Pd(dppf)₂Cl₂ was added. The mixture was stirred at room temperature for 10 min, and then 2 mL of potassium carbonate solution dissolved in water was slowly added. The reaction was carried out at 110 °C for 10 h. After the reaction was completed, the reaction solution was filtered, the solvent was evaporated, and the mixture was stirred and subjected to column chromatography (DCM:MeOH = 50:1) to obtain a pale yellow solid product H in 81.5% yield.

[0092] Synthesis of Compound I:

[0093] Compound H (0.3 g, 0.90 mmol) dissolved in N,N-dimethylformamide was added to a reaction flask. Sodium hydride (0.4 g, 1.8 mmol) was slowly added under ice bath conditions, and the mixture was stirred for 10 min. Bromopropyne (0.15 mL, 1.8 mmol) was added dropwise, and the reaction was allowed to proceed at room temperature for 1 h. After the reaction was complete, the sodium hydride was quenched with water, extracted with ethyl acetate, washed with saturated brine, and dried over anhydrous sodium sulfate. The product was evaporated to dryness to give a yellow solid, product I, in 91% yield.

[0094] Synthesis of Class III compounds:

[0095] 0.2 g (1 eq) of the substituted aniline compound was added to a 50 mL round-bottom flask, along with 2 mL of hydrochloric acid and 2 mL of water. The mixture was stirred at room temperature for 20 min, then cooled to below 0°C on an ice bath. 2 mL of a solution of sodium nitrite dissolved in water (1.1 eq) was added submerged in the mixture, and the mixture was stirred for 30 min. Then, 2 mL of a solution of sodium azide dissolved in water (2 eq) was added dropwise, and the mixture was stirred for 4–8 h. After the reaction was complete, the mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, and evaporated to dryness at low temperature to obtain the azide product.

[0096] Compound I (1 mmol) and aniline azide M3-R (2 mmol) were added to a reaction flask and dissolved in 9 mL of methanol and 3 mL of water. The mixture was stirred at room temperature until dissolved. Copper sulfate pentahydrate aqueous solution (60 mg / mL) was added, and the mixture was stirred for 10 min. Sodium ascorbate solution (32 mg / mL) was then added dropwise, and the reaction was carried out at room temperature (TLC monitoring). A suitable amount of 200-300 mesh silica gel was added as a sample, and the solvent was recovered under reduced pressure. The corresponding compound of formula III was obtained by column chromatography on silica gel, with yields between 50% and 80%.

[0097]

[0098] III-1: 1H NMR (500 MHz, DMSO-d6) δ 8.61 (s, 1H), 8.47 (d, J = 2.6 Hz,1H), 8.20 (td, J = 8.2, 2.7 Hz, 1H), 7.80-7.75 (m, 2H), 7.70 (d, J = 2.2 Hz,1H), 7.63 (dd, J = 8.5, 2.2 Hz, 1H), 7.41 (d, J = 7.3 Hz, 2H), 7.36 (t, J =7.6 Hz, 2H), 7.30-7.25 (m, 1H), 7.22 (dd, J = 8.6, 2.9 Hz, 1H), 7.14-7.08 (m,3H), 5.77 (s, 1H), 5.27 (d, J = 15.3 Hz, 1H), 4.03 (d, J = 15.3 Hz, 1H), 3.81(s, 3H), 3.37 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 161.80, 159.64, 153.77,145.30, 145.18, 144.34, 142.03, 140.19, 138.05, 133.64, 130.49, 129.85,129.49, 128.38, 127.06, 126.67, 125.32, 124.79, 122.38, 122.09, 115.26,114.54, 110.13, 109.83, 60.80, 56.01, 49.06, 30.76. HRMS (ESI) m / z Calculatedfor C 30 H 25 FN6O2 [M+Na] + :539.1921, found:539.1909.

[0099] III-2: 1H NMR (500 MHz, DMSO-d6) δ 8.47 (d, J = 2.6 Hz, 1H), 8.45 (d,J = 2.0 Hz, 1H), 8.20 (td, J = 8.2, 2.7 Hz, 1H), 7.78 (td, J = 7.9, 1.7 Hz,1H), 7.70 (d, J = 2.3 Hz, 1H), 7.63 (dd, J = 8.5, 2.3 Hz, 1H), 7.57-7.53 (m,1H), 7.44-7.38 (m, 3H), 7.35 (t, J = 7.6 Hz, 2H), 7.29-7.24 (m, 1H), 7.22(dd, J = 8.6, 2.9 Hz, 1H), 7.11 (d, J = 8.6 Hz, 1H), 5.80 (s, 1H), 5.23 (d, J= 15.3 Hz, 1H), 4.15-4.10 (m, 2H), 3.16 (d, J = 5.2 Hz, 3H). 13 C NMR (125 MHz,DMSO-d6) δ 155.16, 153.80, 153.17, 145.31, 145.19, 144.24, 142.08, 140.21,138.04, 131.69, 129.88, 129.46, 128.36, 127.09, 126.63, 126.31, 125.99,125.65, 125.27, 124.84, 117.67, 117.52, 114.57, 110.15, 109.85, 61.20, 49.06,30.74. HRMS (ESI) m / z Calculated for C 29 H 22 F2N6O [M+Na] + :531.1721, found:531.1711.

[0100] III-3: 1H NMR (500 MHz, DMSO-d6) δ 8.49 (d, J = 2.6 Hz, 1H), 8.31 (s,1H), 8.22 (td, J = 8.2, 2.6 Hz, 1H), 7.72 (d, J = 2.2 Hz, 1H), 7.65 (dd, J =8.5, 2.2 Hz, 1H), 7.58 (dd, J = 7.8, 1.7 Hz, 1H), 7.52 (ddd, J = 9.3, 7.5,1.7 Hz, 1H), 7.43-7.39 (m, 2H), 7.36 (t, J = 7.6 Hz, 2H), 7.29 (q, J = 7.7,7.2 Hz, 2H), 7.23 (dd, J = 8.6, 2.8 Hz, 1H), 7.17-7.09 (m, 2H), 5.81 (s, 1H),5.25 (d, J = 15.3 Hz, 1H), 4.09 (d, J = 15.3 Hz, 1H), 3.83 (s, 3H), 3.35 (s,3H). 13 C NMR (125 MHz, DMSO-d6) δ 153.81, 151.93, 145.32, 145.20, 143.33,142.04, 140.21, 138.06, 131.13, 129.86, 129.47, 128.35, 127.07, 126.62,126.13, 126.06, 125.23, 124.86, 121.28, 114.56, 113.42, 110.15, 109.85,61.01, 56.53, 49.06, 30.75. HRMS (ESI) m / z Calculated for C 30 H 25 FN6O2 [M+Na] + :543.1921, found:543.1911.

[0101] III-4: 1H NMR (500 MHz, DMSO-d6) δ 8.49 (d, J = 2.6 Hz, 1H), 8.46 (s,1H), 8.21 (td, J = 8.2, 2.7 Hz, 1H), 7.88 (d, J = 2.3 Hz, 1H), 7.85 (d, J =8.6 Hz, 1H), 7.76 (dd, J = 8.6, 2.4 Hz, 1H), 7.69 (d, J = 2.2 Hz, 1H), 7.65(dd, J = 8.5, 2.2 Hz, 1H), 7.43-7.34 (m, 4H), 7.31-7.26 (m, 1H), 7.24 (dd, J= 8.6, 2.8 Hz, 1H), 7.13 (d, J = 8.6 Hz, 1H), 5.77 (s, 1H), 5.26 (d, J = 15.3Hz, 1H), 4.13 (d, J = 15.3 Hz, 1H), 3.32 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ153.79, 145.33, 145.21, 143.59, 141.95, 140.23, 138.05, 137.74, 135.58,135.09, 131.68, 129.90, 129.50, 128.36, 127.12, 126.67, 126.49, 125.23,124.86, 121.39, 118.77, 114.60, 110.18, 60.97, 30.78. HRMS (ESI) m / zCalculated for C 29 H 21 Br2FN6O [M+Na] + :669.0025, found:669.0020.

[0102] III-5: 1H NMR (500 MHz, DMSO-d6) δ 8.76 (s, 1H), 8.48 (d, J = 2.6 Hz,1H), 8.20 (td, J = 8.2, 2.7 Hz, 1H), 7.71 (d, J = 2.2 Hz, 1H), 7.64 (dd, J =8.4, 2.2 Hz, 1H), 7.47-7.34 (m, 4H), 7.29 (t, J = 7.2 Hz, 1H), 7.22 (dd, J =8.6, 2.9 Hz, 1H), 7.17 (s, 2H), 7.12 (d, J = 8.5 Hz, 1H), 5.75 (s, 1H), 5.29(d, J = 15.3 Hz, 1H), 4.05 (d, J = 15.2 Hz, 1H), 3.86 (s, 6H), 3.70 (s, 3H),3.36 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 153.91, 145.30, 144.43, 141.90,140.18, 138.00, 137.65, 132.95, 129.90, 129.51, 128.40, 127.07, 126.65,125.30, 124.76, 122.75, 114.56, 110.12, 109.83, 98.36, 60.65, 56.72. HRMS(ESI) m / z Calculated for C 32 H 29 FN6O4 [M+Na] + :603.2132, found:603.2122.

[0103] III-6: 1H NMR (500 MHz, DMSO-d6) δ 8.80 (s, 1H), 8.47 (d, J = 2.6 Hz,1H), 8.20 (td, J = 8.2, 2.6 Hz, 1H), 8.02 (dd, J = 8.7, 6.1 Hz, 2H), 7.68 (d,J = 2.2 Hz, 1H), 7.64 (dd, J = 8.5, 2.2 Hz, 1H), 7.43-7.33 (m, 4H), 7.28 (t,J = 7.2 Hz, 1H), 7.23 (dd, J = 8.6, 2.9 Hz, 1H), 7.13 (d, J = 8.5 Hz, 1H),5.75 (s, 1H), 5.26 (d, J = 15.4 Hz, 1H), 4.05-4.00 (m, 1H), 3.36-3.34 (m,3H). 13 C NMR (125 MHz, DMSO-d6) δ 153.75, 145.19, 141.96, 140.20, 140.13,137.98, 129.89, 129.51, 128.43, 127.11, 126.67, 125.34, 124.73, 122.92,114.60, 106.13, 105.93, 61.02, 30.76. HRMS (ESI) m / z Calculated for C 29 H 20 F4N6O[M+Na] + :567.1532, found:567.1523.

[0104] III-7: 1H NMR (500 MHz, DMSO-d6) δ 8.49 (s, 2H), 8.22 (td, J = 8.2,2.6 Hz, 1H), 7.74-7.69 (m, 2H), 7.65 (dd, J = 8.5, 2.1 Hz, 1H), 7.48-7.39 (m,4H), 7.36 (t, J = 7.6 Hz, 2H), 7.30-7.27 (m, 1H), 7.24 (dd, J = 8.4, 2.9 Hz,1H), 7.13 (d, J = 8.5 Hz, 1H), 5.85 (s, 1H), 5.21 (d, J = 15.5 Hz, 1H), 4.16(d, J = 15.4 Hz, 1H), 3.37 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 153.80,145.34, 145.22, 144.12, 142.07, 140.23, 140.17, 138.03, 133.66, 129.93,129.47, 128.38, 127.13, 127.08, 126.60, 125.24, 124.85, 114.60, 113.39,113.21, 110.17, 109.86, 61.58, 30.74. HRMS (ESI) m / z Calculated for C 29 H 21 F3N6O[M+Na] + :549.1627, found:549.1613.

[0105] III-8: 1H NMR (500 MHz, DMSO-d6) δ 8.49 (d, J = 4.9 Hz, 2H), 8.21 (td,J = 8.2, 2.7 Hz, 1H), 7.83 (d, J = 2.5 Hz, 1H), 7.80 (d, J = 8.7 Hz, 1H),7.72 (dd, J = 8.7, 2.4 Hz, 1H), 7.70 (d, J = 2.2 Hz, 1H), 7.65 (dd, J = 8.5,2.2 Hz, 1H), 7.42-7.34 (m, 4H), 7.30-7.26 (m, 1H), 7.24 (dd, J = 8.6, 2.8 Hz,1H), 7.13 (d, J = 8.6 Hz, 1H), 5.80 (s, 1H), 5.24 (d, J = 15.4 Hz, 1H), 4.15(d, J = 15.3 Hz, 1H), 3.37 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 153.79,145.33, 145.21, 143.78, 142.01, 140.22, 138.06, 135.83, 133.65, 132.83,132.39, 131.86, 129.91, 129.48, 128.61, 128.36, 127.95, 127.12, 126.66,126.43, 125.23, 124.88, 114.59, 110.17, 109.87, 61.16, 30.76. HRMS (ESI) m / zCalculated for C 29 H 21 Cl2FN6O [M+Na] + :581.1036, found:581.1024.

[0106] III-9: 1H NMR (500 MHz, DMSO-d6) δ 8.75 (s, 1H), 8.48 (d, J = 2.6 Hz,1H), 8.20 (td, J = 8.2, 2.7 Hz, 1H), 7.98-7.91 (m, 2H), 7.74-7.68 (m, 3H),7.64 (dd, J = 8.5, 2.2 Hz, 1H), 7.43-7.34 (m, 4H), 7.30-7.25 (m, 1H), 7.23(dd, J = 8.6, 2.8 Hz, 1H), 7.13 (d, J = 8.6 Hz, 1H), 5.76 (s, 1H), 5.26 (d, J= 15.3 Hz, 1H), 4.05 (d, J = 15.3 Hz, 1H), 3.41 (s, 3H). 13 C NMR (125 MHz,DMSO-d6) δ 153.76, 145.30, 145.18, 144.83, 142.01, 140.18, 140.12, 138.99,138.02, 136.69, 133.63, 129.85, 129.49, 128.39, 127.07, 126.67, 125.33,124.76, 122.39, 122.32, 114.56, 110.14, 109.84, 94.72, 60.90, 30.76. HRMS(ESI) m / z Calculated for C 29 H 22 FIN6O [M+Na] + :638.0782, found:638.0771.

[0107] III-10: 1H NMR (500 MHz, DMSO-d6) δ 8.95 (s, 1H), 8.47 (d, J = 2.7 Hz,1H), 8.45-8.42 (m, 2H), 8.23-8.20 (m, 2H), 8.19 (dd, J = 8.1, 2.7 Hz, 1H),7.68 (d, J = 2.2 Hz, 1H), 7.64 (dd, J = 8.5, 2.2 Hz, 1H), 7.45-7.32 (m, 4H),7.27 (t, J = 7.2 Hz, 1H), 7.23 (dd, J = 8.6, 2.8 Hz, 1H), 7.13 (d, J = 8.6Hz, 1H), 5.77 (s, 1H), 5.28 (d, J = 15.4 Hz, 1H), 4.10 (d, J = 15.4 Hz, 1H), 3.38 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 153.77, 147.10, 145.36, 145.18,142.01, 141.27, 140.19, 140.12, 138.00, 129.88, 129.50, 128.41, 127.09,126.68, 126.01, 125.34, 124.76, 122.99, 120.98, 114.59, 110.15, 109.85,99.99, 61.01, 30.77. HRMS (ESI) m / z Calculated for 29 H 22 FN7O3 [M+Na] + :558.1666, found:558.1658.

[0108] II. Antitumor Activity Test Section

[0109] The novel quinazolinone derivatives provided by this invention possess several significant advantages, and their excellent effects have been verified through systematic biological experiments. First, these compounds exhibit novel structures. Through innovative modification of the C6 and N3 sites of the quinazolinone core, structurally diverse molecules with high targeting specificity are obtained, effectively inhibiting BRD4 protein function. Second, the synthetic method offers significant advantages. Key steps such as Suzuki coupling and click chemistry reactions are characterized by mild conditions, simple operation, and high yields, with most steps exceeding 70% yield. Furthermore, the route is suitable for large-scale preparation, laying a solid foundation for subsequent development.

[0110] In terms of efficacy verification, in vitro and in vivo antitumor activity tests showed that multiple compounds exhibited strong inhibitory activity against breast cancer MCF-7, leukemia HL-60, and gastric cancer HGC-27 cell lines. Some compounds, such as I-11, I-15, and I-18, showed high IC50 values. 50 The activity levels reached the micromolar to nanomolar range, exhibiting superior or equivalent activity to the positive control drug JQ1, and demonstrating good cell selectivity. Further investigation into the mechanism of action focused on the highly active compound I-11: Western blotting experiments demonstrated that this compound dose-dependently inhibited the expression of the key oncogenic protein c-Myc, achieving the same inhibitory effect as 5 μM JQ1 at a concentration of 1.25 μM; scratch assays showed that it significantly inhibited the migration ability of MCF-7 cells; colony formation assays demonstrated that it effectively inhibited cell proliferation in a dose-dependent manner, achieving a 90% inhibition rate at a concentration of 5 μM; flow cytometry analysis showed that I-11 arrested the cell cycle at the G2 / M phase and induced significant apoptosis, with the apoptosis rate increasing to 37.44% at a concentration of 5 μM. These experimental results collectively demonstrate that the compound of the present invention effectively inhibits the proliferation and migration of tumor cells and promotes their apoptosis by regulating the BRD4 / c-Myc signaling pathway. The mechanism of action is clear. In vivo anti-tumor results show that I-11 can effectively inhibit the growth of tumors in vivo (see Figure 1A, B), and there is no significant weight loss in mice, nor is there significant damage or inflammation in organs such as the heart, liver, spleen, lungs and kidneys (see Figure 1C, D), indicating that it has good safety at an intraperitoneal injection dose of 20 mg / kg.

[0111] Example 4: Determination of the inhibitory activity of compounds of formulas I, II, and III against BRD4

[0112] The inhibitory activity of compounds against BRD4 was tested using homogeneous time-resolved fluorescence (HTRF), a method that integrates time-resolved measurement (TR) with fluorescence resonance energy transfer (FRET). The principle of TR-FRET is to use time-resolved fluorescence and energy transfer to detect the proximity between two labeled molecules, thereby reflecting enzyme activity. When a compound inhibits enzyme activity, substrate modification or intermolecular interactions decrease, leading to a reduction in the FRET signal. Measuring this signal change allows for the assessment of the compound's inhibitory ability.

[0113] IC50 analysis of the compound was performed using Graphpad Prism 9.5 software. 50 The LogP values ​​of the compounds were calculated using ChemDraw software, and the specific results are shown in the table below.

[0114]

[0115]

[0116] Note: a Mean of two experiments, the inhibition rate of the compound against BRD4; ND indicates not tested or not calculated; b ChemDraw 21.1 calculation results.

[0117] Experimental methods: First, weigh the target compound and prepare a solution for storage. For inhibition rate determination, replicate tests were performed at a concentration of 2 μM; for IC50... 50 The assay was performed using replicates at concentration gradients of 5 μM, 2.5 μM, 1.25 μM, 0.62 μM, 0.31 μM, 0.15 μM, 0.07 μM, and 0.03 μM, with JQ1 as a positive control compound. The inhibitory effects of different compounds on the two BRD4 protein isoforms, BD1 and BD2, were tested according to the instructions of the BRD4 TR-FRET kit (Cayman Laboratories, USA). In short, 5 μL of the compound with the concentration gradient and 10 μL of europium chloride solution of BRD4-BD1 / BD2 were added to each well of a 384-well plate and incubated at room temperature in the dark for 15 min. Then, 5 μL of BRD4 ligand / APC receptor complex was added to each well and incubated at room temperature in the dark for 1 h. The plate was read using a multi-plate reader, with the excitation wavelength set to 340 nm, and the absorbance values ​​at emission wavelengths of 620 nm and 670 nm were recorded as F670nm / F620nm. The difference in absorbance between the negative control and the compound group, divided by the absorbance value of the negative control, represents the BRD4 inhibition rate. The concentration of the small molecule compound corresponding to a 50% inhibition rate is the half-maximal inhibitory concentration (IC50). 50 ).

[0118] Example 5: Inhibitory effects of compounds of formulas I, II, and III on three cancer cell lines

[0119] The experiment utilized the antiproliferative activity of drug-sensitive breast cancer cells (MCF-7), leukemia cells (HL-60), and gastric cancer cells (HGC-27) with clearly defined sources and pathological classifications to observe the direct growth inhibition or killing effect of the synthesized target compounds on cells in vitro.

[0120] Experimental duration: 48 h

[0121] Inhibition rate % = (OD value of negative control group – OD value of experimental group) / (OD value of negative control group – OD value of blank group) * 100%, IC50 50 The values ​​were calculated using the software GraphPad Prism 9.5.

[0122]

[0123] Experimental steps:

[0124] Tumor cells in the logarithmic growth phase were collected and seeded at 3000-5000 cells / well into 96-well plates and cultured for 24 hours. Cells were observed under an inverted microscope. If the cells were in good condition, 100 μL of the desired compound at different concentrations was added to each well; control wells were treated with only 100 μL of complete culture medium. The plates were then incubated for another 48 hours. After incubation, 20 μL of CCK-8 was added to each well, and incubation continued for 1-4 hours until the color turned orange. The absorbance at 450 nm was then measured.

[0125] Example 6: In vivo antitumor experiment of compound I-11

[0126] All experiments were conducted according to the guidelines and protocols approved by the Henan Provincial Association for Animal Experimentation and Protection. The animals were housed in a sterile environment with sterile food and water under standard laboratory conditions. Female Balb / c nude mice (6 weeks old) were subcutaneously injected with MCF-7 cells into the dorsal region of their right limb to construct a xenograft model. Tumors were allowed to grow to 5 mm in diameter. 3 At approximately 21 days, mice were injected intraperitoneally with a specific dose of the compound for drug intervention, while the control group received an equal volume of drug-free solvent. Mice were randomly divided into three groups (n=5 per group): a blank control group, a compound I-11 group (20 mg / kg), and a positive control JQ-1 group (20 mg / kg). Tumor volume and mouse body weight changes were measured every other day, and tumor size and body weight change curves were established. Furthermore, mice were sacrificed at the observation endpoint, and serum was collected from the orbital bloodstream to detect pathologically relevant serum indicators. Tumors and major organs (heart, liver, spleen, lungs, and kidneys) were also harvested for paraffin sectioning and HE staining to observe histopathological changes and to preliminarily evaluate the dosage safety of the selected molecule.

[0127] The preparation and activity testing of the specific compounds described above fully demonstrate that the quinazolinone derivatives provided by this invention have a series of outstanding advantages, such as efficient and reliable synthesis methods, novel product structures, significant antitumor activity, and certain target selectivity, and have great potential to be developed into novel antitumor drugs.

Claims

1. A quinazolinone derivative, characterized in that, The quinazolinone derivative or its pharmaceutically acceptable salt or solvate has a chemical structure shown in Formula I, Formula II or Formula III: ； In Formulas I, II, and III, X1 and X2 are independently selected from the following groups: ； R is selected from hydrogen atom, deuterium atom, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted deuterated C1-C8 alkyl, substituted or unsubstituted C2-C8 alkenyl, substituted or unsubstituted C1-C8 alkoxy, halogen, amino, nitro, hydroxyl, acyl, cyano, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 5-8 membered heterocyclic group containing 1-3 heteroatoms selected from N, O, S, substituted or unsubstituted 5-8 membered aryl, substituted or unsubstituted group containing 1-3 heteroatoms selected from N, O, S. One of the 5- to 8-membered heteroaryl groups containing O and S heteroatoms; the substitution refers to substitution by one or more substituents selected from the group consisting of: C1-C8 alkyl, halo-C1-C8 alkyl, halogen, amino, nitro, cyano, hydroxyl, C1-C8 alkoxy, halo-C1-C8 alkoxy, hydroxy-C1-C8 alkyl, C3-C8 cycloalkyl, 5- to 8-membered heterocyclic group containing 1 to 3 N, O, and S heteroatoms, 5- to 8-membered aryl, and 5- to 8-membered heteroaryl group containing 1 to 3 N, O, and S heteroatoms.

2. The quinazolinone derivative as described in claim 1, characterized in that: The R is selected from one of the following: substituted or unsubstituted C1-C8 alkoxy groups, substituted or unsubstituted C1-C8 alkyl groups, substituted or unsubstituted C3-C8 cycloalkyl groups, substituted or unsubstituted 5-8 membered heterocyclic groups containing 1-3 heteroatoms selected from N, O, and S, substituted or unsubstituted 5-8 membered aryl groups, and substituted or unsubstituted 5-8 membered heteroaryl groups containing 1-3 heteroatoms selected from N, O, and S; the substitution is one or more substituents selected from the group consisting of: C1-C8 alkyl, halo-C1-C8 alkyl, halogen, amino, nitrate. The following groups are included: alkyl, cyano, hydroxy, hydroxymethyl, hydroxyethyl, mercapto, carboxyl, ester, C1-C6 alkyl monosubstituted amino, C1-C6 alkyl disubstituted amino, C1-C6 alkoxy, C1-C6 alkyl carbonyloxy, C3-C6 cycloalkyl carbonyloxy, 5-8 membered heterocyclic carbonyloxy containing 1-3 heteroatoms selected from N, O, and S, C3-C6 alkyl carbonyl, C3-C6 cycloalkoxy carbonyl, 5-8 membered heterocyclic carbonyloxy containing 1-3 heteroatoms selected from N, O, and S, C1-C6 alkoxyformamide, and C1-C6 alkyl mercapto.

3. The quinazolinone derivative as described in claim 1, characterized in that, The pharmaceutically acceptable salt is an inorganic acid salt, an organic acid salt, or an alkyl sulfonate; the inorganic acid salt includes hydrochloride, hydrobromide, nitrate, sulfate, or phosphate; the organic acid salt includes formate, acetate, propionate, benzoate, maleate, fumarate, succinate, tartrate, or citrate; and the alkyl sulfonate includes methanesulfonate or ethyl sulfonate.

4. The quinazolinone derivative as described in claim 1, characterized in that, The pharmaceutically acceptable solvates are solvates formed by quinazolinone derivatives with water, ethanol, isopropanol, diethyl ether, or acetone.

5. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises any one of the quinazolinone derivatives of claims 1-4, a pharmaceutically acceptable salt or a pharmaceutically acceptable solvate thereof, and one or more pharmaceutically acceptable excipients.

6. Use of the pharmaceutical composition of claim 5 in the preparation of a medicament for treating breast cancer, leukemia, or gastric cancer.

7. The method for preparing the quinazolinone derivative as described in claims 1-4, characterized in that, Includes the following steps: (1) Starting from 2-aminobenzophenone, the key intermediate A of quinazolinone was synthesized by sequentially undergoing bromination, acylation, reduction, cyclization and methylation reactions; (2) Compound C is obtained by reacting intermediate A with compound B via a Suzuki coupling reaction; (3) For compound I: react compound C with an alkyne reagent to introduce an alkyne group to obtain compound D, and then synthesize compound I by a click chemistry reaction of compound D with an azide compound derived from substituted aniline; (4) For compound II: react compound C with an alkyne reagent to introduce an alkyne group to obtain compound E, hydrolyze the ester group of compound E to a carboxyl group to obtain compound F, and then synthesize compound II by condensation reaction of compound F with substituted aniline; (5) For compound III: intermediate A is coupled with compound G via a Suzuki coupling reaction to obtain compound H. Compound H is then reacted with an alkyne reagent to introduce an alkyne group to obtain compound I. Compound I is then synthesized with an azide compound derived from substituted aniline via a click chemistry reaction to obtain compound III. The specific reaction route is as follows: ； 。

Images