WO2024109642A1 - USE OF BENZOAZACYCLIC COMPOUND AS ALLOSTERIC MODULATOR OF β2-ADRENOCEPTOR - Google Patents

Abstract

The present invention belongs to the field of pharmaceutical chemistry, and particularly discloses the use of a benzoazacyclic compound as an allosteric modulator of a β₂-adrenoceptor. The benzoazacyclic compound of the present invention has a structure as shown in formula (I), wherein X is N or CH; Y is N or CH; R₁ is any one or more of H, methyl, methoxy, phenyl, F, Cl, Br, I, CN, NO₂, NH₂, OH and CF₃ at any position on a benzene ring; R₂ is any one of H, methyl, phenyl and benzyl; R₃ is any one of hydrogen and alkyl; and R₄ is any one of H, alkyl, cycloalkyl, phenyl, substituted phenyl and substituted benzyl. The pharmacological results show that most synthesized benzoazacyclic compounds have a good allosteric antagonistic activity on β₂-AR, and can negatively allosterically modulate the functional activity of the receptor.

Description

Application of a benzazine heterocyclic compound as a β2-adrenaline receptor allosteric modulator Technical Field

The invention belongs to the field of medicinal chemistry, and specifically discloses the application of a benzazazepine heterocyclic compound as a β ₂ -adrenergic receptor (β _{2 -AR} ) allosteric modulator.

Background technique

G protein-coupled receptors (GPCRs) are the largest gene family in the human genome, with more than 800 members. GPCRs are closely related to a wide range of physiological functions, including development, immunity, hormone regulation, and neuronal activity. Typically, GPCRs convert extracellular stimuli into intracellular responses by activating heterotrimeric G proteins composed of Ga, Gb, and Gg subunits. (Cell Reports 2023, 42: 113173).

GPCRs signals are transmitted through two pathways: G protein-dependent and G protein-independent, producing different biological effects (Journal of Medicinal Chemistry 2018, 61(22): 9841-9878.). The former often involves the activation of G proteins, while the latter often triggers the recruitment of arrestins proteins.

G-protein-coupled receptors (GPCRs) are currently the most studied and most important drug targets, and are involved in almost all physiological, pathological and pharmacological processes in humans. So far, most GPCRs drugs on the market are their orthosteric ligands, which have many adverse reactions and poor specificity. GPCRs allosteric modulators have become a research hotspot in recent years due to their huge potential advantages. β ₂ -AR antagonists are very classic GPCRs drugs, playing an important role in the treatment of diseases such as heart failure, hypertension, coronary heart disease, arrhythmia, angina pectoris, etc., and are the cornerstone of the treatment of cardiovascular diseases. Therefore, the development and design of β ₂ -AR allosteric antagonists are of great significance for the treatment of various cardiovascular diseases.

In 2017, our research group and collaborators reported a small molecule negative allosteric modulator compound 15 (Cmpd-15), which is the first intracellular allosteric antagonist of β ₂ -AR (Proc. Natl. Acad. Sci. USA, 2017, 114: 1708-1713). However, since Cmpd-15 is a peptide compound with poor water solubility and low biological activity, the relative instability of its structure may affect its drugability. Later, our research group transformed Cmpd-15 into an N-benzyl-substituted pyrazole/phenylalanine derivative (see Formula 1; R' = substituted benzyl) through skeleton transition and structural simplification strategies, which improved the allosteric regulatory activity on β ₂ -AR and greatly improved the solubility (CN 115894373 A). However, the activity of these compounds is still not ideal, and the structure is relatively complex (containing chiral phenylalanine fragments), which is difficult to synthesize. Therefore, we tried to transform the structures of these compounds into benzopyrazole amide compounds (see the benzopyrazole general formula in the middle of Formula 2). Then, using the skeleton hopping strategy, we designed and synthesized benzimidazole and indole derivatives (see the two structural general formulas on the right side of Formula 2). Then, we tested the functional activity of all the new compounds synthesized through the G protein-dependent signaling pathway (GloSensor ^TM cAMP Assay Promega), and screened out some compounds with significantly better allosteric antagonistic activity than the lead compound Cmpd-15. Moreover, the structures of these compounds are obviously simple and can be obtained by only one to three steps of synthesis reaction.

Summary of the invention

The present invention uses various substituted 3-carboxylic acid benzazazepines and various amines as raw materials, and synthesizes a series of various substituted benzopyrazole amides, benzimidazole amides and indole amide derivatives through amide coupling reaction. The GloSensor cAMP accumulation experiment confirms that most of the synthesized compounds are allosteric antagonists of β ₂ -AR, and the activity of some of the compounds is significantly higher than that of the lead compound Cmpd-15. The present invention may provide a solid foundation for the creation of new drugs for cardiovascular, asthma and cancer diseases.

The present invention provides a benzene nitrogen heterocyclic compound, the structural formula of the benzene nitrogen heterocyclic compound is as follows:

X＝N,CH;

Y＝N,CH;

R ₁ =H, methyl, methoxy, phenyl, F, Cl, Br, I, CN, NO ₂ , NH ₂ , OH, CF ₃ ;

R ₂ =H, methyl, phenyl, benzyl;

R ₃ =H, alkyl;

R ₄ =H, alkyl, cycloalkyl, phenyl, substituted phenyl, substituted benzyl.

Furthermore, the benzazine heterocyclic compound is a benzimidazole amide compound, and its general structural formula is shown in Formula I:

Wherein R ₁ is H, methyl, methoxy, phenyl, F, Cl, Br, I, CN, NO ₂ , NH ₂ , OH, CF ₃ ; R ₃ is H, alkyl; R ₄ is H, phenyl, bromophenyl, fluorophenyl.

Furthermore, the benzazine heterocyclic compound is a benzopyrazole amide compound, and its general structural formula is shown in Formula II:

Wherein R ₁ is H, methyl, methoxy, phenyl, F, Cl, Br, I, CN, NO ₂ , NH ₂ , OH, CF ₃ ; R ₂ is H, methyl, methoxy, phenyl, benzyl; R ₃ is H, alkyl; R ₄ is H, phenyl, bromophenyl, fluorophenyl.

Furthermore, the benzazine heterocyclic compound is an indoleamide compound, and its general structural formula is shown in Formula III:

Wherein R ₁ is H, methyl, methoxy, phenyl, F, Cl, Br, I, CN, NO ₂ , NH ₂ , OH, CF ₃ ; R ₃ is H, alkyl; R ₄ is H, phenyl, bromophenyl, fluorophenyl.

Furthermore, the benzazazepine heterocyclic compound is an allosteric modulator of β ₂ -adrenergic receptor (β _{2 -AR} ).

The present invention also provides a method for synthesizing the above-mentioned benzazazepine heterocyclic compound, which specifically comprises the following steps: a simple amide coupling reaction is carried out between 3-carboxylic acid benzazazepine and various amines, firstly dissolving 3-carboxylic acid benzazazepine and an activator in a solvent, adding different types of amines under ice bath, then adding an acid binding agent and an amide coupling agent and continuing to stir to room temperature; the activator is 1-hydroxy-7-azabenzotriazole (HOAT), the acid binding agent is N-methylmorpholine (NMM), and the amide coupling agent is 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI); the molar ratio of 3-carboxylic acid benzazazepine: HOAT: amine: NMM: EDCI is 1:1.2-1.5:1-2:0.6-0.75:1.2-1.5.

The synthesis steps of benzyl azoheterocyclic compounds are as follows:

1. The synthetic route of benzyl azoheterocyclic compounds is as follows:

(1) The synthetic route of benzimidazole amide compounds is as follows:

(2) The synthetic route of benzopyrazole amide compounds is as follows:

(3) The synthetic route of indoleamide compounds is as follows:

2. The synthesis steps of benzyl azoheterocyclic compounds are as follows:

(1) The general synthesis steps of benzimidazole amide compounds are as follows:

1) Dissolve o-phenylenediamine (1 eq) and glycolic acid (4 eq) in 6N dilute hydrochloric acid and reflux for 2-3 h. Post-treatment: Cool to room temperature, adjust pH to neutral with concentrated ammonia in an ice bath, and filter to obtain benzimidazole alcohol.

2) Dissolve alcohol 2 (1 eq) and NaOH (2 eq) in water, stir at 80°C for 2 h, add KMnO ₄ (1.5 eq) in batches, reflux for 10 h, cool and filter, take the filtrate, add dilute hydrochloric acid in an ice bath to adjust the pH to 4, filter, and obtain benzimidazole acid.

3) Dissolve benzimidazole acid and activator in solvent, add different types of amines under ice bath, then add acid binder and amide coupling agent and continue stirring to room temperature. Wherein, the solvent is N,N-dimethylformamide, the activator is 1-hydroxy-7-azabenzotriazole (HOAT), the acid binder is N-methylmorpholine (NMM), and the amide coupling agent is 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI). Dissolve benzimidazole acid (1eq) and HOAT (1.2eq) in DMF, stir for 10min, and add different amines. Then, add NMM (0.7eq) at 0℃, stir for 10min, add EDCI (1.2eq), keep stirring at 0℃ for 1.5h, and react at room temperature for 12h. Spin dry DMF, extract, pass column to obtain benzazaheterocyclic amide compounds.

(2) The general synthesis steps of benzopyrazole amide compounds are as follows:

At room temperature, 3-carboxylic acid benzopyrazole is added to DMF, and then amine and HOAT are added. After stirring at room temperature for 10 minutes, N-methylmorpholine (NMM) is added at 0°C. After stirring at ⁰ °C for 10 minutes, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) is added, and then the mixture is reacted at 0°C for 9 hours. The mixture is extracted with ethyl acetate, and the organic phase is concentrated and separated by column chromatography to obtain benzazaheterocyclic amide compounds.

(3) The specific synthesis method steps of indoleamides are as follows:

Indolecarboxylic acid was dissolved in N,N-dimethylformamide (DMF) in a round-bottom flask, N-hydroxy-7-azabenzotriazole (HOAT) was added and reacted for 10 minutes, then amine and N-methylmorpholine (NMM) were added under ice bath conditions and reacted for 10 minutes, and then 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCl) was added and reacted at room temperature for 4 hours. After the reaction was completed by TLC monitoring, ethyl acetate and water were extracted three times to remove DMF, the organic phase was collected and dried over anhydrous sodium sulfate, and then the solvent was removed by rotary evaporation, and the obtained solid crude product was purified by silica gel column chromatography, and the crude product of the concentrated column chromatography was recrystallized with an appropriate amount of dichloromethane to obtain indoleamide compounds.

Table 1-1. List of compounds synthesized from benzimidazole amides

Table 1-2. List of compounds synthesized from benzopyrazoles

Table 1-3. List of compounds synthesized from indoleamides

The benzazazepine heterocyclic compound of the present invention is used for preparing beta ₂ -adrenaline receptor allosteric antagonist.

The present invention has the following beneficial effects:

The present invention discloses that benzopyrazole amide compounds are a new type of allosteric antagonist of β ₂ -AR, and some of the compounds have significantly higher activity than the lead compound Cmpd-15. Moreover, these compounds have simple structures and can be obtained by only one organic reaction. The present invention may provide a solid foundation for the creation of new drugs for cardiovascular, asthma and cancer diseases.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG1 is a graph showing a dose-response curve of ISO mediated by benzimidazole amide derivative A08;

FIG2 is a graph showing the ISO dose-response curve mediated by the benzopyrazole amide derivative B08;

FIG3 is a graph showing the ISO dose-response curve mediated by the indoleamide derivative C01;

FIG4 is a graph showing a dose-response curve of ISO mediated by indoleamide derivative C15;

FIG5 is a graph showing the ISO dose-response curve mediated by the indoleamide derivative C17.

Detailed ways

1. Preparation of benzimidazole amide compounds

Embodiment 1:

Preparation of 5-dimethyl-1H-benzo[d]imidazole-2-carboxamide(A01)

Step 1: Dissolve 4-methylbenzene-1,2-diamine (1 g, 8.2 mmol) and glycolic acid (2.5 g, 32.8 mmol) in 6N HCl (41 mL) under ice bath, and stir under reflux at 120 ^° C for 12 h. After the reaction, cool to room temperature, add concentrated ammonia water under ice bath to adjust pH to neutral, filter, and wash with water to obtain a pink solid compound (1.1 g, yield 79%).

Step 2: Dissolve the crude product of step 1 (1.1 g, 6.7 mmol) in 34 ml of water, add solid NaOH (0.54 g, 13.5 mmol), heat and stir at 80 degrees for 2 h. Then add KMnO4 (1.6 g, 10.2 mmol) in batches and reflux for 10 h. After the reaction is completed, cool to room temperature, adjust the pH to 4 with 6N HCl in an ice bath, filter, wash the solid with water, and column chromatography (dichloromethane: methanol: formic acid = 10: 1: 0.1) to obtain a milky white solid compound 5-methyl-1H-benzimidazole-2-carboxylic acid (766 mg, yield 60%).

Step 3: Dissolve 5-methyl-1H-benzimidazole-2-carboxylic acid (200 mg, 1.13 mmol) and HOAT (185 mg, 1.36 mmol) in DMF (6 mL), stir for 10 min, add compound methylamine hydrochloride (154 mg, 2.26 mmol), add N-methylmorpholine (0.092 mL, 0.79 mmol) under ice bath and stir for 10 min, add EDCI (261 mg, 1.36 mmol), keep stirring at 0 ° C for 1 h, and react at room temperature for 12 h. After the reaction is completed, spin-dry DMF, extract with ethyl acetate, wash the organic phase with saturated brine, concentrate column chromatography (petroleum ether: ethyl acetate = 3: 1), and obtain 150 mg of white solid product with a yield of 70%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 13.14 (s, 1H), 8.92 (s, 1H), 7.57 (d, J＝8.3 Hz, 1H), 7.31 (s, 1H), 7.06 (d, J＝8.4 Hz, 1H), 2.84 (s, 3H), 2.40 (s, 3H). MS (ESI, m/z): 190 [M+1] ⁺ .

Embodiment 2:

Preparation of N-cyclopentyl-5-methyl-1H-benzo[d]imidazole-2-carboxamide(A02)

The preparation method is the same as that of Example 1, except that cyclopentylamine is used in place of methylamine hydrochloride in step 3, and a white solid is finally obtained with a yield of 68%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ13.10 (s, 1H), 8.75 (d, J=8.0 Hz, 1H), 7.57-7.31 (m, 2H), 7.09 (s, 1H), 4.27 (m, 1H), 2.41 (s, 3H), 1.91-1.82 (m, 2H), 1.73-1.49 (m, 6H). MS (ESI, m/z): 244 [M+1] ⁺ .

Embodiment 3:

Preparation of N-cyclopentyl-5-methoxy-1H-benzo[d]imidazole-2-carboxamide(A03)

The preparation method is the same as that of Example 1, except that 4-methoxybenzene-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and cyclopentylamine is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 65%. ¹ H NMR (400MHz, DMSO-d ₆ ): δ13.08 (s, 1H), 8.67 (d, J＝8.1Hz, 1H), 7.57-6.91 (m, 3H), 4.26 (q, J＝7.3Hz, 1H), 3.79 (s, 3H), 1.91-1.82 (m, 2H), 1.72-1.50 (m, 6H). MS (ESI, m/z): 260 [M+1] ⁺ .

Embodiment 4:

Preparation of N-cyclohexyl-5-methyl-1H-benzo[d]imidazole-2-carboxamide(A04)

The preparation method is the same as that of Example 1, except that cyclopentylamine is used in place of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 65%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ13.19 (s, 1H), 8.71 (d, J=8.6 Hz, 1H), 7.66 (d, J=8.3 Hz, 1H), 7.40 (s, 1H), 7.16 (d, J=8.3 Hz, 1H), 3.88-3.84 (m, 1H), 1.82-1.78 (m, 4H), 1.66 (s, 1H), 1.51-1.48 (m, 2H), 1.37-1.34 (m, 2H), 1.17-1.14 (m, 1H). MS (ESI, m/z): 258 [M+1] ⁺ .

Embodiment 5:

Preparation of N-cyclohexyl-5-methoxy-1H-benzo[d]imidazole-2-carboxamide(A05)

The preparation method is the same as that of Example 1, except that 4-methoxybenzene-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and cyclohexylamine is used instead of methylamine hydrochloride in step 3, and the yield is 62%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ13.10 (s, 1H), 8.55 (d, J＝8.5 Hz, 1H), 7.54 (s, 1H), 6.97-6.89 (m, 2H), 3.78 (s, 3H), 3.38 (s, 1H), 1.80-1.69 (m, 4H), 1.60-1.57 (m, 1H), 1.48-1.25 (m, 4H), 1.15-1.06 (m, 1H). MS (ESI, m/z): 274 [M+1] ⁺ .

Embodiment 6:

Preparation of N-phenyl-1H-benzo[d]imidazole-2-carboxamide(A06)

The preparation method is the same as that of Example 1, except that benzene-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and aniline is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 55%. ¹ H NMR (400MHz, DMSO-d ₆ ): δ13.46 (s, 1H), 10.92 (s, 1H), 7.95 (d, J＝8.0Hz, 2H), 7.70 (s, 2H), 7.36 (d, J＝29.6Hz, 4H), 7.13 (d, J＝14.8Hz, 1H). MS (ESI, m/z): 238 [M+1] ⁺ .

Embodiment 7:

Preparation of 5-methyl-N-phenyl-1H-benzo[d]imidazole-2-carboxamide(A07)

The preparation method is the same as that of Example 1, except that aniline is used in place of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 55%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ13.45 (s, 1H), 11.09 (d, J=10.5 Hz, 1H), 8.25 (s, 1H), 7.93 (d, J=7.3 Hz, 1H), 7.68-7.57 (m, 1H), 7.49-7.38 (m, 1H), 7.33 (d, J=7.5 Hz, 2H), 7.19-7.12 (m, 1H), 2.43 (s, 3H). MS (ESI, m/z): 252 [M+1] ⁺ .

Embodiment 8:

Preparation of 5-methoxy-N-phenyl-1H-benzo[d]imidazole-2-carboxamide(A08)

The preparation method is the same as that of Example 1, except that 4-methoxybenzene-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and aniline is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 58%. ¹ HNMR (400MHz, DMSO-d ₆ ): δ13.32 (s, 1H), 10.79 (s, 1H), 7.92 (d, J＝8.0Hz, 2H), 7.61 (s, 1H), 7.37 (t, J＝7.7Hz, 2H), 7.12 (t, J＝7.4Hz, 1H), 7.05 (s, 1H), 6.96 (d, J＝9.1Hz, 1H), 3.82 (s, 3H). MS (ESI, m/z): 268 [M+1] ⁺ .

Embodiment 9:

Preparation of 5-fluoro-N-phenyl-1H-benzo[d]imidazole-2-carboxamide(A09)

The preparation method is the same as that of Example 1, except that 4-fluoro-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and aniline is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ9.67 (t, J＝6.4 Hz, 1H), 7.67-7.63 (m, 2H), 7.55 (s, 1H), 7.44 (d, J＝7.9 Hz, 1H), 7.36-7.29 (m, 3H), 4.49 (d, J＝6.2 Hz, 2H). MS (ESI, m/z): 256 [M+1] ⁺ .

Embodiment 10:

Preparation of 5-chloro-N-phenyl-1H-benzo[d]imidazole-2-carboxamide(A10)

The preparation method is the same as that of Example 1, except that 4-chloro-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and aniline is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. ¹ H NMR (400MHz, DMSO-d ₆ ): δ13.66 (s, 1H), 10.99 (s, 1H), 7.94 (d, J＝8.0Hz, 2H), 7.68 (s, 2H), 7.39-7.35 (m, 3H), 7.13 (t, J＝7.4Hz, 1H). MS (ESI, m/z): 272 [M+1] ⁺ .

Embodiment 11:

Preparation of 5-bromo-N-phenyl-1H-benzo[d]imidazole-2-carboxamide(A11)

The preparation method is the same as that of Example 1, except that 4-bromo-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and aniline is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 55%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ13.53 (s, 1H), 11.00 (s, 1H), 7.93 (d, J＝8.0 Hz, 3H), 7.64 (s, 1H), 7.48 (d, J＝8.6 Hz, 1H), 7.38 (t, J＝7.7 Hz, 2H), 7.14 (t, J＝7.4 Hz, 1H). MS (ESI, m/z): 316 [M+1] ⁺ .

Embodiment 12:

Preparation of 5-nitro-N-phenyl-1H-benzo[d]imidazole-2-carboxamide(A12)

The preparation method is the same as that of Example 1, except that 4-nitrobenzene-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and aniline is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ14.10 (s, 1H), 11.10 (s, 1H), 8.52 (s, 1H), 8.21 (d, J＝8.9 Hz, 1H), 7.93 (d, J＝7.9 Hz, 2H), 7.82 (d, J＝8.3 Hz, 1H), 7.39 (d, J＝7.7 Hz, 2H), 7.15 (t, J＝7.4 Hz, 1H). MS (ESI, m/z): 283 [M+1] ⁺ .

Embodiment 13:

Preparation of N-(3-bromophenyl)-1H-benzo[d]imidazole-2-carboxamide(A13)

The preparation method is the same as that of Example 1, except that benzene-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and m-bromoaniline is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. ¹ H NMR (400MHz, DMSO-d ₆ ): δ13.55 (s, 1H), 11.14 (s, 1H), 8.26 (s, 1H), 7.96-7.93 (m, 1H), 7.81 (d, J＝8.0 Hz, 1H), 7.59 (d, J＝8.0 Hz, 1H), 7.37-7.31 (m, 4H). MS (ESI, m/z): 316 [M+1] ⁺ .

Embodiment 14:

Preparation of N-(3-bromophenyl)-5-methyl-1H-benzo[d]imidazole-2-carboxamide(A14)

The preparation method is the same as that of Example 1, except that 4-methylbenzene-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and m-bromoaniline is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. ¹ H NMR (400MHz, DMSO-d ₆ ): δ13.32 (s, 1H), 10.84 (s, 1H), 7.93 (d, J＝7.9Hz, 2H), 7.37 (t, J＝7.8Hz, 3H), 7.13 (t, J＝7.3Hz, 2H), 2.45 (s, 3H). MS (ESI, m/z): 330 [M+1] ⁺ .

Embodiment 15:

Preparation of N-(3-bromophenyl)-5-methoxy-1H-benzo[d]imidazole-2-carboxamide(A15)

The preparation method is the same as that of Example 1, except that 4-methoxybenzene-1,2-diamine is used in place of 4-methylbenzene-1,2-diamine in step 1, and m-bromoaniline is used in place of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ13.35 (s, 1H), 11.02 (s, 1H), 8.25 (s, 1H), 7.93 (d, J＝11.3 Hz, 1H), 7.69 (s, 1H), 7.32 (d, J＝23.3 Hz, 2H), 6.99 (d, J＝17.6 Hz, 2H), 3.82 (s, 3H). MS (ESI, m/z): 346 [M+1] ⁺ .

Embodiment 16:

Preparation of N-(3-bromophenyl)-5-fluoro-1H-benzo[d]imidazole-2-carboxamide(A16)

The preparation method is the same as that of Example 1, except that 4-fluoro-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and m-bromoaniline is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ13.59 (s, 1H), 11.13 (s, 1H), 8.25 (s, 1H), 7.94-7.91 (m, 1H), 7.72-7.69 (m, 1H), 7.45 (d, J＝9.2 Hz, 1H), 7.36-7.33 (m, 2H), 7.23-7.18 (m, 1H). MS (ESI, m/z): 333 [M+1] ⁺ .

Embodiment 17:

Preparation of N-(3-bromophenyl)-5-chloro-1H-benzo[d]imidazole-2-carboxamide(A17)

The preparation method is the same as that of Example 1, except that 4-chloro-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and m-bromoaniline is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. ¹ H NMR (400MHz, DMSO-d ₆ ): δ13.67 (s, 1H), 11.19 (s, 1H), 8.23 (s, 1H), 7.93 (d, J＝6.4Hz, 1H), 7.71 (s, 2H), 7.38-7.33 (m, 3H). MS (ESI, m/z): 349 [M+1] ⁺ .

Embodiment 18:

Preparation of 5-bromo-N-(3-bromophenyl)-1H-benzo[d]imidazole-2-carboxamide(A18)

The preparation method is the same as that of Example 1, except that 4-bromo-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and m-bromoaniline is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. ¹ H NMR (400MHz, DMSO-d ₆ ): δ13.70 (s, 1H), 11.21 (s, 1H), 7.94-7.91 (m, 2H), 7.55 (s, 1H), 7.34-7.32 (m, 2H). MS (ESI, m/z): 395 [M+1] ⁺ .

Embodiment 19:

Preparation of N-(3-bromophenyl)-5-nitro-1H-benzo[d]imidazole-2-carboxamide(A19)

The preparation method is the same as that of Example 1, except that 4-nitro-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and m-bromoaniline is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. ¹ H NMR (400MHz, DMSO-d ₆ ): δ14.12 (s, 1H), 11.30 (s, 1H), 8.51 (s, 1H), 8.22-8.19 (m, 2H), 7.94-7.80 (m, 2H), 7.36-7.33 (m, 2H). MS (ESI, m/z): 360 [M+1] ⁺ .

Embodiment 20:

Preparation of N-(3-bromobenzyl)-5-methyl-1H-benzo[d]imidazole-2-carboxamide(A20)

The preparation method is the same as that of Example 1, except that 4-methyl-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and m-bromobenzylamine is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. ¹ H NMR (400MHz, DMSO-d ₆ ): δ13.15 (s, 1H), 9.57 (s, 1H), 7.55 (s, 1H), 7.50 (dd, J＝8.4, 4.4 Hz, 1H), 7.44 (d, J＝7.8 Hz, 1H), 7.36-7.27 (m, 3H), 7.12 (s, 1H), 4.48 (d, J＝6.4 Hz, 2H), 2.42 (s, 3H). MS (ESI, m/z): 344 [M+1] ⁺ .

Embodiment 21:

Preparation of N-(3-bromobenzyl)-5-methoxy-1H-benzo[d]imidazole-2-carboxamide(A21)

The preparation method is the same as that of Example 1, except that 4-methoxy-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and m-bromobenzylamine is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. ¹ H NMR (400MHz, DMSO-d ₆ ): δ13.15 (s, 1H), 9.52 (s, 1H), 7.59-6.90 (m, 7H), 4.49 (s, 2H), 3.79 (s, 3H). MS (ESI, m/z): 360 [M+1] ⁺ .

Embodiment 22:

Preparation of N-(3-bromobenzyl)-5-fluoro-1H-benzo[d]imidazole-2-carboxamide(A22)

The preparation method is the same as that of Example 1, except that 4-fluoro-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and m-bromobenzylamine is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ13.39 (s, 1H), 9.63 (t, J＝6.4 Hz, 1H), 7.66-7.61 (m, 1H), 7.55 (s, 1H), 7.45 (d, J＝7.8 Hz, 1H), 7.36-7.27 (m, 3H), 7.18 (t, J＝8.9 Hz, 1H), 4.49 (d, J＝6.3 Hz, 2H). MS (ESI, m/z): 348 [M+1] ⁺ .

Embodiment 23:

Preparation of N-(3-bromobenzyl)-5-chloro-1H-benzo[d]imidazole-2-carboxamide(A23)

The preparation method is the same as that of Example 1, except that 4-chloro-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and m-bromobenzylamine is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. ¹ H NMR (400MHz, DMSO-d ₆ ): δ12.98 (s, 1H), 9.67 (t, J＝6.4Hz, 1H), 7.70–7.61 (m, 2H), 7.55 (s, 1H), 7.45 (d, J＝6.3Hz, 1H), 7.36-7.27 (m, 3H), 4.49 (d, J＝6.2Hz, 2H). MS (ESI, m/z): 363 [M+1] ⁺ .

Embodiment 24:

Preparation of 5-bromo-N-(3-bromobenzyl)-1H-benzo[d]imidazole-2-carboxamide(A24)

The preparation method is the same as that of Example 1, except that 4-bromo-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and m-bromobenzylamine is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ9.70 (t, J=6.4 Hz, 1H), 7.82 (s, 1H), 7.59 (d, J=8.7 Hz, 1H), 7.55 (s, 1H), 7.44 (dd, J=8.7, 1.9 Hz, 2H), 7.35 (d, J=7.8 Hz, 1H), 7.28 (t, J=7.7 Hz, 1H), 4.49 (d, J=6.4 Hz, 2H). MS (ESI, m/z): 409 [M+1] ⁺ .

Embodiment 25:

Preparation of N-(3-bromobenzyl)-5-nitro-1H-benzo[d]imidazole-2-carboxamide(A25)

The preparation method is the same as that of Example 1, except that 4-nitro-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and m-bromobenzylamine is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ14.01 (s, 1H), 9.85 (t, J=6.4 Hz, 1H), 8.52 (s, 1H), 8.20 (dd, J=9.0, 2.3 Hz, 1H), 7.80 (d, J=9.1 Hz, 1H), 7.56 (s, 1H), 7.45 (d, J=7.9 Hz, 1H), 7.36 (d, J=7.8 Hz, 1H), 7.30 (t, J=7.7 Hz, 1H), 4.51 (d, J=6.4 Hz, 2H). MS (ESI, m/z): 375 [M+1] ⁺ .

Embodiment 26:

Preparation of N-(3-fluorophenyl)-5-methyl-1H-benzo[d]imidazole-2-carboxamide(A26)

The preparation method is the same as that of Example 1, except that 4-methyl-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and m-fluoroaniline is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ13.37 (d, J=7.9 Hz, 1H), 11.11 (d, J=10.1 Hz, 1H), 7.88 (d, J=11.8 Hz, 1H), 7.79 (d, J=8.2 Hz, 1H), 7.69-7.58 (m, 1H), 7.47-7.37 (m, 2H), 7.20-7.13 (m, 1H), 6.98-6.94 (m, 1H), 2.45 (s, 3H). MS (ESI, m/z): 270 [M+1] ⁺ .

Embodiment 27:

Preparation of N-(3-fluorophenyl)-5-methoxy-1H-benzo[d]imidazole-2-carboxamide(A27)

The preparation method is the same as that of Example 1, except that 4-methoxy-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and m-fluoroaniline is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. ¹ H NMR (400MHz, DMSO-d ₆ ): δ13.36 (s, 1H), 11.04 (s, 1H), 7.90-7.86 (m, 1H), 7.79-7.77 (m, 1H), 7.68-7.66 (d, J＝9.0Hz, 1H), 7.43-7.37 (m, 1H), 7.00 (s, 1H), 6.98-6.92 (m, 2H), 3.81 (s, 3H). MS (ESI, m/z): 286 [M+1] ⁺ .

Embodiment 28:

Preparation of N-(3-chlorophenyl)-5-methyl-1H-benzo[d]imidazole-2-carboxamide(A28)

The preparation method is the same as that of Example 1, except that 4-methyl-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and m-chloroaniline is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. ¹ H NMR (400MHz, DMSO-d ₆ ): δ13.35 (s, 1H), 11.02 (s, 1H), 8.25 (s, 1H), 7.93 (d, J＝11.3 Hz, 1H), 7.69 (s, 1H), 7.32 (d, J＝23.3 Hz, 2H), 6.99 (d, J＝17.6 Hz, 2H), 3.82 (s, 3H). MS (ESI, m/z): 286 [M+1] ⁺ .

Embodiment 29:

Preparation of N-(3-chlorophenyl)-5-methoxy-1H-benzo[d]imidazole-2-carboxamide(A29)

The preparation method is the same as that of Example 1, except that 4-chloromethoxy-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and m-chloroaniline is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. ¹ H NMR (400MHz, DMSO-d ₆ ): δ13.34 (s, 1H), 11.03 (s, 1H), 8.10 (t, J＝2.1Hz, 1H), 7.91-7.88 (m, 1H), 7.67 (d, J＝8.9Hz, 1H), 7.39 (t, J＝8.1Hz, 1H), 7.19-7.17 (m, 1H), 7.0-6.9 (m, 2H), 3.82 (s, 3H). MS (ESI, m/z): 302 [M+1] ⁺ .

Embodiment 30:

Preparation of N-(3,5-dibromophenyl)-5-methyl-1H-benzo[d]imidazole-2-carboxamide(A30)

The preparation method is the same as that of Example 1, except that 4-methyl-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and 3,5-dibromoaniline is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. MS (ESI, m/z): 409 [M+1] ⁺ .

Embodiment 31:

Preparation of N-(3,5-dibromophenyl)-5-methoxy-1H-benzo[d]imidazole-2-carboxamide(A31)

The preparation method is the same as that of Example 1, except that 4-methoxy-1,2-diamine is used instead of 4-methylbenzene-1,2-diamine in step 1, and 3,5-dibromoaniline is used instead of methylamine hydrochloride in step 3 to obtain a white solid with a yield of 52%. ¹ H NMR (400MHz, DMSO-d ₆ ): δ13.35 (s, 1H), 11.17 (s, 1H), 8.24 (s, 2H), 7.67 (d, J＝8.9Hz, 1H), 7.55-7.55 (m, 1H), 6.99 (d, J＝2.5Hz, 1H), 6.96-6.93 (m, 1H), 3.82 (s, 3H). MS (ESI, m/z): 425 [M+1] ⁺ .

2. Preparation of benzopyrazole amide compounds

Embodiment 32:

Synthesis of (S)-N-(3-(3-bromophenyl)-1-(methylamino)-1-oxopropan-2-yl)-1-phenyl-1H-indazole-3-carboxamide (B1)

At room temperature, 47.7 mg (0.2 mmol, 1 eq) of N-phenylindazole-3-carboxylic acid was added to N,N-dimethylformamide (2 ml), and then 51.4 mg (0.2 mmol, 1 eq) of 2-amino-3-(3-bromophenyl)-N-methylpropionamide and 41 mg (0.3 mmol, 1.5 eq) of N-hydroxy-7-azabenzotriazole were added. After stirring at room temperature for 10 minutes, N-methylmorpholine (18 μL, 0.15 mmol, 0.75 eq) was added at 0 ^{°C. After stirring at 0°} C for 10 minutes, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (58 mg, 0.3 mmol, 1.5 eq) was added. The mixture was reacted at 0°C for 9 hours, extracted with ethyl acetate, and the organic phase was concentrated and separated by column chromatography to obtain a white solid B1 (14 mg, yield 15%). ¹ H NMR (400 MHz, CDCl ₃ ): δ8.38 (d, J=8.2 Hz, 1H), 7.74-7.72 (m, 3H), 7.60-7.56 (m, 2H), 7.49-7.43 (m, 3H), 7.38-7.34 (m, 2H), 7.26-7.23 (m, 2H), 7.16 (t, J=7.8 Hz, 1H), 4.87 (t, J=7.0 Hz, 1H), 3.22 (dd, J=6.6, 3.4 Hz, 2H), 2.75 (s, 3H); HRMS (ESI) m/z: calculated for C ₂₄ H ₂₁ BrN ₄ NaO ₂ ⁺ [M+Na] ⁺ 499.0740, found 499.0742.

Embodiment 33:

Synthesis of N-(3-fluorophenyl)-1-phenyl-1H-indazole-3-carboxamide(B2)

The preparation method is the same as that of Example 32, except that 2-amino-3-(3-bromophenyl)-N-methylpropionamide is replaced by m-fluoroaniline, and compound B2 is finally obtained as a white solid with a yield of 78%. ¹ H NMR (400 MHz, CDCl ₃ ) δ9.01 (s, 1H), 8.52 (d, J=8.2 Hz, 1H), 7.80-7.71 (m, 4H), 7.63-7.58 (m, 2H), 7.52-7.46 (m, 2H), 7.41-7.36 (m, 2H), 7.34-7.29 (m, 1H), 6.87-6.82 (m, 1H).

Embodiment 34:

Synthesis of N-(3-chlorophenyl)-1-phenyl-1H-indazole-3-carboxamide(B3)

The preparation method is the same as that of Example 32, except that 2-amino-3-(3-bromophenyl)-N-methylpropionamide is replaced by m-chloroaniline, and compound B3 is finally obtained as a white solid with a yield of 67%. ¹ H NMR (400 MHz, CDCl ₃ ) δ8.97 (s, 1H), 8.51 (d, J=8.2 Hz, 1H), 7.93 (t, J=2.0 Hz, 1H), 7.76-7.71 (m, 3H), 7.63-7.57 (m, 3H), 7.52-7.46 (m, 2H), 7.41-7.37 (m, 1H), 7.29 (t, J=8.0 Hz, 1H), 7.13-7.10 (m, 1H).

Embodiment 35:

Synthesis of N-(3-bromophenyl)-1-phenyl-1H-indazole-3-carboxamide(B4)

The preparation method is the same as that of Example 32, except that 2-amino-3-(3-bromophenyl)-N-methylpropionamide is replaced by m-bromoaniline, and compound B4 is finally obtained as a white solid with a yield of 68%. ¹ H NMR (400 MHz, CDCl ₃ ): δ8.99 (s, 1H), 8.56 (d, J=8.2 Hz, 1H), 8.11 (t, J=1.9 Hz, 1H), 7.81-7.76 (m, 3H), 7.71-7.63 (m, 3H), 7.57-7.50 (m, 2H), 7.46-7.42 (m, 1H), 7.31-7.28 (m, 2H).

Embodiment 36:

Synthesis of (S)-1-benzyl-N-(3-(3-bromophenyl)-1-(methylamino)-1-oxopropan-2-yl)-1H-indazole-3-carboxamide(B5)

The preparation method is the same as that of Example 32, except that N-phenylindazole-3-carboxylic acid is replaced by N-benzylindazole-3-carboxylic acid, and compound B5 is finally obtained as a white solid with a yield of 68%. ¹ H NMR (400 MHz, CDCl ₃ ) δ8.26 (d, J=8.2 Hz, 1H), 7.59 (d, J=8.3 Hz, 1H), 7.48 (s, 1H), 7.39-7.19 (m, 10H), 7.13 (t, J=7.8 Hz, 1H), 6.37 (s, 1H), 5.59 (s, 2H), 4.91 (q, J=7.3 Hz, 1H), 3.27-3.18 (m, 2H), 2.74 (d, J=4.6 Hz, 3H). ¹³ C NMR (100 MHz, CDCl ₃ ):δ171.2,162.7,141.0,139.4,136.8,135.9,132.6,130.3,130.2,129.0,128.3,128.2,127.4,127.2,123.2,123.1,122.7,122.5,109.9,54.2,53.8,38.1,26.4.

Embodiment 37:

Synthesis of N-(3-bromophenyl)-1-methyl-1H-indazole-3-carboxamide(B6)

The preparation method is the same as that of Example 32, except that N-phenylindazole-3-carboxylic acid is replaced by N-methylindazole-3-carboxylic acid, and 2-amino-3-(3-bromophenyl)-N-methylpropionamide is replaced by m-bromoaniline, and compound B6 is finally obtained as a white solid with a yield of 90%. ¹ H NMR (400 MHz, (CD ₃ ) ₂ SO): δ10.59 (s, 1H), 8.27 (t, J＝2.0 Hz, 1H), 8.24-8.21 (m, 1H), 7.89-7.86 (m, 1H), 7.80-7.77 (m, 1H), 7.53-7.48 (m, 1H), 7.35-7.26 (m, 3H), 4.20 (s, 3H). ¹³ C NMR (100 MHz, (CD ₃ ) ₂ SO):δ160.9,141.2,140.6,136.5,130.6,126.9,126.1,122.9,122.6,122.5,121.7,121.5,119.1,110.7,36.1.HRMS(ESI)m/z:calcd for C ₁₅ H ₁₂ BrN ₃ NaO ⁺ [M+Na] ⁺ 352.0056,found 352.0059.

Embodiment 38:

Synthesis of (S)-N-(3-(3-bromophenyl)-1-(methylamino)-1-oxopropan-2-yl)-1H-indazole-3-carboxamide(B7)

The preparation method is the same as that of Example 32, except that N-phenylindazole-3-carboxylic acid is replaced by indazole-3-carboxylic acid, and finally compound B7 is obtained as a white solid with a yield of 71%. ¹ H NMR (400 MHz, CDCl ₃ )δ14.41(s,1H),9.95(d,J＝8.3Hz,1H),8.24(d,J＝5.1Hz,1H),8.12(d,J＝8.2Hz,1H),7.76(d,J＝8.5Hz,1H),7.48(t,J＝7.6Hz,1H),7.37(t,J＝1.8Hz,1H),7.29(t,J＝7.6Hz,1H),7.23(d,J＝7.7Hz,1H),7.18(dd,J＝8.0,1.9Hz,1H),6.99(t,J＝7.8Hz,1H),5.32(q,J＝8.0Hz,1H),3.26-3.13(m,2H),2.95(d,J＝4.6Hz,3H). ¹³ C NMR(100MHz,CDCl ₃ )δ174.1,163.9,141.4,139.1,137.7,132.5,130.2,130.1,127.8,127.1,122.9,122.5,121.9,111.2,55.4,37.6,26.6.HRMS(ESI)m/z:calcd for C ₁₈ H ₁₇ BrN ₄ NaO ₂ ⁺ [M+Na] ⁺ 423.0427,found 423.0429.

Embodiment 39:

Synthesis of N-(3-bromophenyl)-1H-indazole-3-carboxamide(B8)

The preparation method is the same as that of Example 32, except that N-phenylindazole-3-carboxylic acid is replaced by indazole-3-carboxylic acid, and 2-amino-3-(3-bromophenyl)-N-methylpropionamide is replaced by m-bromoaniline, to finally obtain compound B8 as a white solid with a yield of 78%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ13.87 (s, 1H), 10.59 (s, 1H), 8.27 (s, 1H), 8.23 (d, J=8.1 Hz, 1H), 7.89 (dt, J=7.8, 1.8 Hz, 1H), 7.68 (d, J=8.4 Hz, 1H), 7.48-7.45 (m, 1H), 7.33-7.26 (m, 3H); ¹³ C NMR (100 MHz, DMSO-d ₆ ): δ161.3, 141.4, 140.6, 138.0, 130.6, 126.9, 126.0, 122.6, 122.5, 121.8, 121.5, 119.0, 111.0. HRMS (ESI) m/z: calcd for C ₁₄ H ₁₀ BrN ₃ NaO ⁺ [M+Na] ⁺ 337.9900, found 337.9900.

Embodiment 40:

Synthesis of N-(3-fluorophenyl)-1H-indazole-3-carboxamide(B9)

The preparation method is the same as that of Example 32, except that N-phenylindazole-3-carboxylic acid is replaced by indazole-3-carboxylic acid, and 2-amino-3-(3-bromophenyl)-N-methylpropionamide is replaced by m-fluoroaniline, to finally obtain compound B9 as a white solid with a yield of 63%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ13.86 (s, 1H), 10.61 (s, 1H), 8.24 (d, J=8.2 Hz, 1H), 7.91-7.88 (m, 1H), 7.75 (d, J=8.2 Hz, 1H), 7.68 (d, J=8.4 Hz, 1H), 7.47 (t, J=7.4 Hz, 1H), 7.38 (q, J=7.8 Hz, 1H), 7.31 (t, J=7.6 Hz, 1H), 6.94-6.89 (m, 1H). ¹³ C NMR (100 MHz, DMSO-d ₆ ):δ163.3,161.3,160.9,141.4,140.8,140.7,138.0,130.3,130.2,126.9,122.6,121.8,121.5,116.0,116.0,111.0,110.0,109.7,107.0,106.8.HRMS(ESI)m/z:calcd for C ₁₄ H ₁₀ FN ₃ NaO ⁺ [M+Na] ⁺ 278.0700,found 278.0705.

Embodiment 41:

Synthesis of N-(3-chlorophenyl)-1H-indazole-3-carboxamide(B10)

The preparation method is the same as that of Example 32, except that N-phenylindazole-3-carboxylic acid is replaced by indazole-3-carboxylic acid, and 2-amino-3-(3-bromophenyl)-N-methylpropionamide is replaced by m-chloroaniline, to finally obtain compound B10 as a white solid with a yield of 36%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ13.87 (s, 1H), 10.61 (s, 1H), 8.24 (d, J=8.0 Hz, 1H), 8.13 (s, 1H), 7.86 (dd, J=8.1, 2.1 Hz, 1H), 7.68 (d, J=8.4 Hz, 1H), 7.47 (t, J=7.4 Hz, 1H), 7.37 (t, J=8.1 Hz, 1H), 7.31 (t, J=7.6 Hz, 1H), 7.15-7.13 (m, 1H); ¹³ C NMR (100 MHz, DMSO-d ₆ ):δ161.3,141.4,140.5,138.0,133.0,130.3,126.9,123.1,122.6,121.8,121.5,119.6,118.6,111.0.HRMS(ESI)m/z:calcd for C ₁₄ H ₁₀ ClN ₃ NaO ⁺ [M+Na] ⁺ 294.0404,found 294.0409.

Embodiment 42:

Synthesis of N-(m-tolyl)-1H-indazole-3-carboxamide(B11)

The preparation method is the same as that of Example 32, except that N-phenylindazole-3-carboxylic acid is replaced by indazole-3-carboxylic acid, and 2-amino-3-(3-bromophenyl)-N-methylpropionamide is replaced by m-methylaniline, to finally obtain compound B11 as a light yellow solid with a yield of 76%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ13.87 (s, 1H), 10.61 (s, 1H), 8.24 (d, J=8.0 Hz, 1H), 8.13 (s, 1H), 7.86 (dd, J=8.1, 2.1 Hz, 1H), 7.68 (d, J=8.4 Hz, 1H), 7.47 (t, J=7.4 Hz, 1H), 7.37 (t, J=8.1 Hz, 1H), 7.31 (t, J=7.6 Hz, 1H), 7.15-7.13 (m, 1H); ¹³ C NMR (100 MHz, DMSO-d ₆ ):δ161.3,141.4,140.5,138.0,133.0,130.3,126.9,123.1,122.6,121.8,121.5,119.6,118.6,111.0.HRMS(ESI)m/z:calcd for C ₁₄ H ₁₀ ClN ₃ NaO ⁺ [M+Na] ⁺ 294.0404,found 294.0409.

Embodiment 43:

Synthesis of N-(3,5-dibromophenyl)-1H-indazole-3-carboxamide(B12)

The preparation method is the same as that of Example 32, except that N-phenylindazole-3-carboxylic acid is replaced by indazole-3-carboxylic acid, and 2-amino-3-(3-bromophenyl)-N-methylpropionamide is replaced by 3,5-dibromoaniline, to finally obtain compound B12 as a white solid with a yield of 9%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 13.93 (s, 1H), 10.75 (s, 1H), 8.24 (d, J = 1.8 Hz, 2H), 8.22 (d, J = 8.1 Hz, 1H), 7.68 (d, J = 8.4 Hz, 1H), 7.52 (d, J = 1.6 Hz, 1H), 7.47 (t, J = 7.4 Hz, 1H), 7.31 (t, J = 7.6 Hz, 1H). ¹³ C NMR (100 MHz, DMSO-d ₆ ) δ 161.4, 141.7, 141.4, 137.6, 127.9, 126.9, 122.8, 122.3, 121.8, 121.4, 121.4, 111.1.

Embodiment 44:

Synthesis of N-benzyl-1H-indazole-3-carboxamide(B13)

The preparation method is the same as that of Example 32, except that N-phenylindazole-3-carboxylic acid is replaced by indazole-3-carboxylic acid, and 2-amino-3-(3-bromophenyl)-N-methylpropionamide is replaced by benzylamine, to finally obtain compound B13 as a white solid with a yield of 93%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ13.64 (s, 1H), 9.02 (t, J=6.4 Hz, 1H), 8.19 (d, J=8.1 Hz, 1H), 7.62 (d, J=8.4 Hz, 1H), 7.43-7.39 (m, 1H), 7.37-7.35 (m, 2H), 7.33-7.29 (m, 2H), 7.26-7.20 (m, 2H), 4.51 (d, J=6.4 Hz, 2H); ¹³ C NMR (100 MHz, DMSO-d ₆ ):δ162.4,141.1,140.0,138.3,128.3,127.3,126.7,126.5,122.1,121.6,121.6,110.7,41.9.HRMS(ESI)m/z:calcd for C ₁₅ H ₁₃ N ₃ NaO ⁺ [M+Na] ⁺ 274.0951,found 274.0957.

Embodiment 45:

Synthesis of N-methyl-1H-indazole-3-carboxamide(B14)

The preparation method is the same as that of Example 32, except that N-phenylindazole-3-carboxylic acid is replaced by indazole-3-carboxylic acid, and 2-amino-3-(3-bromophenyl)-N-methylpropionamide is replaced by methylamine, to finally obtain compound B14 as a white solid with a yield of 86%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 13.56 (s, 1H), 8.38 (d, J = 4.9 Hz, 1H), 8.19 (d, J = 8.1 Hz, 1H), 7.60 (d, J = 8.4 Hz, 1H), 7.42-7.38 (m, 1H), 7.23 (t, J = 7.6 Hz, 1H), 2.82 (d, J = 4.7 Hz, 3H). ¹³ C NMR (100 MHz, DMSO-d ₆ ) δ 162.9, 141.1, 138.5, 126.5, 122.0, 121.7, 121.5, 110.7, 25.6.

Embodiment 46:

Synthesis of N-isopropyl-1H-indazole-3-carboxamide(B15)

The preparation method is the same as that of Example 32, except that N-phenylindazole-3-carboxylic acid is replaced by indazole-3-carboxylic acid, and 2-amino-3-(3-bromophenyl)-N-methylpropionamide is replaced by isopropylamine, to finally obtain compound B15 as a white solid with a yield of 83%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 13.55 (s, 1H), 8.19 (d, J = 7.9 Hz, 1H), 8.11 (d, J = 8.3 Hz, 1H), 7.60 (d, J = 8.4 Hz, 1H), 7.42-7.38 (m, 1H), 7.22 (t, J = 7.9 Hz, 1H), 4.24-4.12 (m, 1H), 1.19 (d, J = 6.6 Hz, 6H). ¹³ C NMR (100 MHz, DMSO-d ₆ ) δ 161.4, 141.1, 138.5, 126.5, 122.0, 121.7, 121.6, 110.7, 40.2, 22.4.

Embodiment 47:

Synthesis of N-cyclohexyl-1H-indazole-3-carboxamide(B16)

The preparation method is the same as that of Example 32, except that N-phenylindazole-3-carboxylic acid is replaced by indazole-3-carboxylic acid, and 2-amino-3-(3-bromophenyl)-N-methylpropionamide is replaced by cyclohexylamine, to finally obtain compound B16 as a white solid with a yield of 81%. ¹ H NMR (400 MHz, CD ₃ OD): δ8.21 (d, J＝8.2 Hz, 1H), 7.57-7.55 (m, 1H), 7.43-7.39 (m, 1H), 7.26-7.22 (m, 1H), 3.96-3.90 (m, 1H), 2.01-1.98 (m, 2H), 1.84-1.80 (m, 2H), 1.70-1.65 (m, 1H), 1.50-1.35 (m, 4H), 1.30-1.23 (m, 1H). HRMS (ESI) m/z: calcd for C ₁₄ H ₁₇ N ₃ NaO ⁺ [M+Na] ⁺ 266.1264, found 266.1271.

Embodiment 48:

Synthesis of 5-bromo-N-(3-bromophenyl)-1H-indazole-3-carboxamide(B17)

The preparation method is the same as that of Example 32, except that N-phenylindazole-3-carboxylic acid is replaced by 5-bromoindazole-3-carboxylic acid, and 2-amino-3-(3-bromophenyl)-N-methylpropionamide is replaced by m-bromoaniline, and compound B17 is finally obtained as a white solid with a yield of 36%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ14.07 (s, 1H), 10.66 (s, 1H), 8.37 (d, J=1.8 Hz, 1H), 8.25 (t, J=2.0 Hz, 1H), 7.87 (dt, J=7.7, 1.8 Hz, 1H), 7.67 (d, J=8.8 Hz, 1H), 7.59 (dd, J=8.8, 1.9 Hz, 1H), 7.34-7.26 (m, 2H); ¹³ C NMR (100 MHz, DMSO-d ₆ ):δ160.8,140.4,140.1,137.5,130.6,129.7,126.2,123.6,123.4,122.6,121.5,119.1,115.2,113.3.HRMS(ESI)m/z:calcd for C ₁₄ H ₉ Br ₂ N ₃ NaO ⁺ [M+Na] ⁺ 415.9004,found 415.9001.

Embodiment 49:

Synthesis of 5-bromo-N-phenyl-1H-indazole-3-carboxamide(B18)

The preparation method is the same as that of Example 32, except that N-phenylindazole-3-carboxylic acid is replaced by 5-bromoindazole-3-carboxylic acid, and 2-amino-3-(3-bromophenyl)-N-methylpropionamide is replaced by aniline, to finally obtain compound B18 as a white solid with a yield of 44%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 14.02 (s, 1H), 10.44 (s, 1H), 8.38 (d, J = 1.8 Hz, 1H), 7.90 (d, J = 7.6 Hz, 2H), 7.67 (d, J = 8.8 Hz, 1H), 7.58 (dd, J = 8.9, 1.9 Hz, 1H), 7.35 (t, J = 7.7 Hz, 2H), 7.10 (t, J = 7.4 Hz, 1H). ¹³ C NMR (100 MHz, DMSO-d ₆ ) δ 160.6, 140.1, 138.8, 137.8, 129.6, 128.6, 123.7, 123.6, 123.4, 120.4, 115.0, 113.2.

Embodiment 50:

Synthesis of 5-methyl-N-phenyl-1H-indazole-3-carboxamide(B19)

The preparation method is the same as that of Example 32, except that N-phenylindazole-3-carboxylic acid is replaced by 5-methylindazole-3-carboxylic acid, and 2-amino-3-(3-bromophenyl)-N-methylpropionamide is replaced by aniline, and compound B19 is finally obtained as a yellow solid with a yield of 80%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ13.68 (s, 1H), 10.30 (s, 1H), 8.03 (s, 1H), 7.92-7.90 (m, 2H), 7.56 (d, J＝8.6 Hz, 1H), 7.35 (t, J＝7.5 Hz, 2H), 7.29 (dd, J＝8.6, 1.6 Hz, 1H), 7.10-7.06 (m, 1H), 2.45 (s, 3H); ¹³ C NMR (100 MHz, DMSO-d ₆ ):δ161.1,140.0,139.0,137.7,131.5,128.8,128.6,123.4,122.2,120.5,120.2,110.6,21.2.HRMS(ESI)m/z:calcd for C ₁₅ H ₁₃ N ₃ NaO ⁺ [M+Na] ⁺ 274.0951,found 274.0957.

Embodiment 51:

Synthesis of 6-bromo-N-phenyl-1H-indazole-3-carboxamide(B20)

The preparation method is the same as that of Example 32, except that N-phenylindazole-3-carboxylic acid is replaced by 6-bromoindazole-3-carboxylic acid, and 2-amino-3-(3-bromophenyl)-N-methylpropionamide is replaced by aniline, to finally obtain compound B20 as a white solid with a yield of 83%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ13.93 (s, 1H), 10.42 (s, 1H), 8.16 (d, J=8.6 Hz, 1H), 7.91 (d, J=5.7 Hz, 2H), 7.88 (s, 1H), 7.43 (dd, J=8.6, 1.6 Hz, 1H), 7.35 (t, J=7.7 Hz, 2H), 7.09 (t, J=7.4 Hz, 1H); ¹³ C NMR (100 MHz, DMSO-d ₆ ) δ160.6, 142.1, 138.8, 138.7, 128.6, 125.7, 123.6, 123.4, 120.8, 120.4, 120.3, 113.6. HRMS (ESI) m/z: calculated for C ₁₄ H ₁₀ BrN ₃ NaO ⁺ [M+Na] ⁺ 337.9900, found 337.9906.

Embodiment 52:

Synthesis of 5-chloro-N-phenyl-1H-indazole-3-carboxamide (B21)

The preparation method is the same as that of Example 32, except that N-phenylindazole-3-carboxylic acid is replaced by 5-chloroindazole-3-carboxylic acid, and 2-amino-3-(3-bromophenyl)-N-methylpropionamide is replaced by aniline, to finally obtain compound B21 as a white solid with a yield of 75%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ14.02 (s, 1H), 10.43 (s, 1H), 8.22 (d, J=2.1 Hz, 1H), 7.90 (d, J=8.0 Hz, 2H), 7.72 (d, J=8.9 Hz, 1H), 7.48 (dd, J=8.8, 2.0 Hz, 1H), 7.35 (t, J=7.7 Hz, 2H), 7.10 (t, J=7.4 Hz, 1H). ¹³ C NMR (100 MHz, DMSO-d ₆ ): δ160.6, 139.9, 138.8, 138.0, 128.7, 127.2, 127.0, 123.6, 122.7, 120.5, 120.4, 112.9.

3. Preparation of indoleamide derivatives

Embodiment 53:

Synthesis of 6-bromo-N-phenyl-1H-indole-3-carboxamide(C ₀₁ )

At room temperature, 150 mg (0.6 mmol, 1 eq) of 6-bromo-1H-indole-3-carboxylic acid was added to N,N-dimethylformamide (5 mL), and then N-hydroxy-7-azabenzotriazole (102 mg, 0.75 mmol, 1.2 eq) was added. After reacting for 10 min, aniline (57 μL, 0.44 mmol, 1 eq) was added, and N-methylmorpholine (50 μL) was added at 0°C for 10 min, and then 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (144 mg, 0.75 mmol, 1.2 eq) was added, and reacted at room temperature for 4 h. After the reaction was completed as monitored by TLC, it was extracted with ethyl acetate, and the organic phase was concentrated and separated by column chromatography to obtain a solid crude product, which was then recrystallized from an appropriate amount of dichloromethane to obtain the target compound C ₀₁ as a white solid with a yield of 76%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 11.85 (s, 1H), 9.79 (s, 1H), 8.33 (d, J=3.0 Hz, 1H), 8.13 (d, J=8.5 Hz, 1H), 7.77-7.74 (m, 2H), 7.69 (d, J=1.8 Hz, 1H), 7.35-7.27 (m, 3H), 7.06-7.02 (m, 1H).

Embodiment 54:

Synthesis of 6-bromo-N-(3-bromobenzyl)-1H-indole-3-carboxamide(C ₀₂ )

The preparation method is the same as that of Example 53, except that aniline is replaced by 3-bromobenzylamine, and compound C ₀₂ is finally obtained as a white solid with a yield of 50%. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 11.72 (s, 1H), 8.61 (t, J = 6.0 Hz, 1H), 8.10 (d, J = 8.4 Hz, 2H), 7.65 (d, J = 1.9 Hz, 1H), 7.52 (s, 1H), 7.43 (d, J = 7.8 Hz, 1H), 7.34 (d, J = 7.6 Hz, 1H), 7.30 (d, J = 7.7 Hz, 1H), 7.25 (dd, J = 8.5, 1.8 Hz, 1H), 4.47 (d, J = 6.0 Hz, 2H). ¹³ C NMR (101 MHz, DMSO-d ₆ ): δ 164.3, 143.3, 137.1, 130.5, 129.9, 129.5, 128.8, 126.4, 125.3, 123.4, 122.8, 121.7, 114.7, 114.6, 110.4, 41.4. HRMS (ESI, m/z): Calcd. for C ₁₆ H ₁₂ Br ₂ N ₂ O[M+Na] ⁺ 428.9208, found: 428.9219.

Embodiment 55:

Synthesis of N-benzyl-6-bromo-1H-indole-3-carboxamide(C ₀₃ )

The preparation method is the same as that of Example 53, except that aniline is replaced by benzylamine, and compound C ₀₃ is finally obtained as a white solid with a yield of 70%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.70 (s, 1H), 8.57 (t, J = 6.0 Hz, 1H), 8.13–8.11 (m, 2H), 7.65 (d, J = 1.8 Hz, 1H), 7.36–7.30 (m, 4H), 7.26–7.20 (m, 2H), 4.48 (d, J = 6.0 Hz, 2H). ¹³ C NMR (101 MHz, DMSO-d ₆ ) δ 164.2, 140.4, 137.1, 128.7, 128.3, 127.3, 126.7, 125.4, 123.4, 122.9, 114.7, 114.5, 110.7, 41.9. HRMS (ESI, m/z): Calcd. for C ₁₆ H ₁₃ BrN ₂ O[M+Na] ⁺ 351.0103,found:351.0104.

Embodiment 56:

Synthesis of 6-bromo-N-(3-chlorophenyl)-1H-indole-3-carboxamide(C ₀₄ )

The preparation method is the same as that of Example 53, except that aniline is replaced by 3-chloroaniline, and compound C ₀₄ is finally obtained as a white solid with a yield of 29%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.91 (s, 1H), 9.96 (s, 1H), 8.33 (s, 1H), 8.12 (d, J = 8.5 Hz, 1H), 7.98 (t, J = 2.0 Hz, 1H), 7.70 (d, J = 1.8 Hz, 1H), 7.66 (dd, J = 8.0, 2.0 Hz, 1H), 7.36 (t, J = 8.0 Hz, 1H), 7.30 (dd, J = 8.5, 1.8 Hz, 1H), 7.10 (dd, J = 8.0, 2.1 Hz, 1H). ¹³ C NMR (101 MHz, DMSO-d ₆ )δ163.0,141.2,137.2,133.0,130.3,130.0,125.5,123.8,122.8,122.4,119.1,118.0,115.0,114.8,110.3.

Embodiment 57:

Synthesis of 6-bromo-N-(3-iodophenyl)-1H-indole-3-carboxamide(C ₀₅ )

The preparation method is the same as that of Example 53, except that aniline is replaced by 3-iodoaniline, and compound C ₀₅ is finally obtained as a white solid with a yield of 6%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.89 (s, 1H), 9.86 (s, 1H), 8.32 (s, 1H), 8.25 (t, J = 1.8 Hz, 1H), 8.12 (d, J = 8.5 Hz, 1H), 7.76 (dd, J = 8.1, 2.1 Hz, 1H), 7.69 (d, J = 1.8 Hz, 1H), 7.39 (d, J = 7.9 Hz, 1H), 7.29 (dd, J = 8.5, 1.8 Hz, 1H), 7.13 (t, J = 8.0 Hz, 1H). ¹³ C NMR (101 MHz, DMSO-d ₆ )δ162.9,141.2,137.1,131.2,130.7,129.9,127.7,125.5,123.8,122.8,118.8,115.0,114.7,110.3,94.5.

Embodiment 58:

Synthesis of 6-bromo-N-(m-tolyl)-1H-indole-3-carboxamide(C ₀₆ )

The preparation method is the same as that of Example 53, except that aniline is replaced by 3-methylaniline, and compound C ₀₆ is finally obtained as a white solid with a yield of 7%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.84 (s, 1H), 9.71 (s, 1H), 8.32 (s, 1H), 8.13 (d, J = 8.5 Hz, 1H), 7.68 (d, J = 1.8 Hz, 1H), 7.61 (s, 1H), 7.55 (dd, J = 8.1, 2.1 Hz, 1H), 7.28 (dd, J = 8.5, 1.8 Hz, 1H), 7.20 (t, J = 7.8 Hz, 1H), 6.86 (d, J = 7.5 Hz, 1H). ¹³ C NMR (101 MHz, DMSO-d ₆ )δ162.8,139.6,137.7,137.1,129.5,128.5,125.6,123.6,123.5,122.8,120.3,117.0,114.9,114.7,110.7,21.3.

Embodiment 59:

Synthesis of 6-bromo-N-(4-bromophenyl)-1H-indole-3-carboxamide(C ₀₇ )

The preparation method is the same as that of Example 53, except that aniline is replaced by 4-bromoaniline, to finally obtain compound C ₀₇ as a white solid with a yield of 9%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.88 (s, 1H), 9.91 (s, 1H), 8.32 (s, 1H), 8.12 (d, J = 8.5 Hz, 1H), 7.76–7.74 (m, 2H), 7.69 (d, J = 1.7 Hz, 1H), 7.52–7.50 (m, 2H), 7.29 (dd, J = 8.6, 1.8 Hz, 1H). ¹³ C NMR (101 MHz, DMSO-d ₆ ) δ 162.9, 139.1, 137.1, 131.4, 129.8, 125.5, 123.8, 122.8, 121.6, 115.0, 114.7, 114.3, 110.4.

Embodiment 60:

Synthesis of 6-bromo-N-cyclopentyl-1H-indole-3-carboxamide(C ₀₈ )

The preparation method is the same as that of Example 53, except that aniline is replaced by cyclopentylamine, and compound C ₀₈ is finally obtained as a white solid with a yield of 69%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.65 (s, 1H), 8.08 (dd, J = 5.6, 2.8 Hz, 2H), 7.81 (d, J = 7.3 Hz, 1H), 7.62 (d, J = 1.8 Hz, 1H), 7.22 (dd, J = 8.6, 1.9 Hz, 1H), 4.27-4.18 (m, 1H), 1.91-1.82 (m, 2H), 1.73-1.62 (m, 2H), 1.57-1.45 (m, 4H). ¹³ C NMR (101 MHz, DMSO-d ₆ )δ164.0,137.0,128.6,125.5,123.3,123.0,114.6,114.5,111.0,50.3,32.5,23.7.HRMS(ESI,m/z):Calcd.for C ₁₄ H ₁₅ BrN ₂ O[M+Na] ⁺ 329.0260,found:329.0254.

Example 61:

Synthesis of 6-bromo-N-cyclohexyl-1H-indole-3-carboxamide(C ₀₉ )

The preparation method is the same as that of Example 53, except that aniline is replaced by cyclohexylamine, and compound C ₀₉ is finally obtained as a white solid with a yield of 36%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.62 (s, 1H), 8.09–8.07 (m, 2H), 7.69 (d, J=7.9 Hz, 1H), 7.61 (s, 1H), 7.22 (d, J=8.5 Hz, 1H), 3.81–3.73 (m, 1H), 1.85-1.82 (m, 2H), 1.74-1.68 (m, 2H), 1.60 (d, J=12.6 Hz, 1H), 1.35-1.22 (m, 4H), 1.17–1.08 (m, 1H). ¹³ C NMR (101 MHz, DMSO-d ₆ )δ163.3,136.9,128.4,125.4,123.1,122.8,114.5,114.3,111.0,47.4,32.8,25.4,25.0.HRMS(ESI,m/z):Calcd.for C ₁₅ H ₁₇ BrN ₂ O[M+Na] ⁺ 343.0416,found:343.0412.

Embodiment 62:

Synthesis of 6-bromo-N-methyl-1H-indole-3-carboxamide(C ₁₀ )

The preparation method is the same as that of Example 53, except that aniline is replaced by methylamine hydrochloride, to finally obtain compound C ₁₀ as a white solid with a yield of 66%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.64 (s, 1H), 8.08 (d, J = 8.5 Hz, 1H), 7.98 (s, 1H), 7.95 (d, J = 4.8 Hz, 1H), 7.62 (d, J = 1.8 Hz, 1H), 7.23 (dd, J = 8.5, 1.8 Hz, 1H), 2.76 (d, J = 4.5 Hz, 3H). 13C NMR (101 MHz, DMSO-d 6 ) δ 164.8, 137.0, 128.3, 125.2, 123.3, 122.8, 114.6, 114.5, 111.0, 25.6. HRMS (ESI, m/z): Calcd. for C ₁₀ H ₉ BrN ₂ O [M + Na] ⁺ 274.9790,found:274.9789.

Embodiment 63:

Synthesis of 6-bromo-N,N-dimethyl-1H-indole-3-carboxamide(C ₁₁ )

The preparation method is the same as that of Example 53, except that aniline is replaced by dimethylamine hydrochloride, and compound C ₁₁ is finally obtained as a white solid with a yield of 54%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.68 (s, 1H), 7.76–7.73 (m, 2H), 7.61 (s, 1H), 7.21 (d, J=8.5 Hz, 1H), 3.07 (s, 6H). ¹³ C NMR (101 MHz, DMSO-d ₆ ) δ 165.7, 136.5, 128.8, 125.8, 123.0, 122.6, 114.5, 114.3, 110.0. HRMS (ESI, m/z): Calcd. for C ₁₁ H ₁₁ BrN ₂ O [M+Na] ⁺ 288.9947, found: 288.9944.

Embodiment 64:

Synthesis of N-(3-bromophenyl)-1H-indole-3-carboxamide(C ₁₂ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by indole-3-carboxylic acid, and aniline is replaced by 3-bromoaniline, to finally obtain compound C ₁₂ as a white solid with a yield of 51%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.90 (s, 1H), 8.83 (dd, J = 4.6, 1.4 Hz, 1H), 8.76–8.73 (m, 2H), 7.95 (dd, J = 7.1, 1.8 Hz, 1H), 7.67 (d, J = 4.5 Hz, 1H), 7.65 (d, J = 4.2 Hz, 1H), 7.36–7.28 (m, 2H). ¹³ C NMR (101 MHz, DMSO-d ₆ ) δ 159.9, 152.6, 150.7, 140.5, 136.8, 136.5, 134.5, 129.8, 125.6, 123.7, 122.8, 121.7, 120.0, 113.4, 99.0.

Embodiment 65:

Synthesis of N-phenyl-1H-indole-3-carboxamide(C ₁₃ )

The preparation method is the same as that of Example 32, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by indole-3-carboxylic acid, and compound C ₁₃ is finally obtained as a white solid with a yield of 8%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.74 (s, 1H), 9.72 (s, 1H), 8.30 (s, 1H), 8.20 (d, J = 7.7 Hz, 1H), 7.78 (d, J = 8.0 Hz, 2H), 7.47 (d, J = 7.8 Hz, 1H), 7.33 (t, J = 7.8 Hz, 2H), 7.21-7.13 (m, 2H), 7.04 (t, J = 7.4 Hz, 1H).

Embodiment 66:

Synthesis of N-(3-bromophenyl)-6-fluoro-1H-indole-3-carboxamide(C ₁₄ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 6-fluoro-1H-indole-3-carboxylic acid, and aniline is replaced by 3-bromoaniline, to finally obtain compound C ₁₄ as a light yellow solid with a yield of 17%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.84 (s, 1H), 9.90 (s, 1H), 8.31 (s, 1H), 8.16 (dd, J = 8.8, 5.7 Hz, 1H), 8.12 (t, J = 2.0 Hz, 1H), 7.74-7.71 (m, 1H), 7.32-7.27 (m, 2H), 7.24-7.21 (m, 1H), 7.05-7.00 (m, 1H). ¹³ C NMR (101 MHz, DMSO-d ₆ )δ163.2,160.4,158.0,141.4,136.3,136.2,130.6,129.7,129.7,125.2,123.15,122.2,122.1,121.9,121.5,118.3,110.3,109.5,109.3,98.4,98.1.

Embodiment 67:

Synthesis of 6-fluoro-N-phenyl-1H-indole-3-carboxamide(C ₁₅ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 6-fluoro-1H-indole-3-carboxylic acid, and compound C ₁₅ is finally obtained as a light yellow solid with a yield of 70%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.80 (s, 1H), 9.76 (s, 1H), 8.31 (d, J = 2.9 Hz, 1H), 8.18 (dd, J = 8.8, 5.7 Hz, 1H), 7.77 (d, J = 7.2 Hz, 1H), 7.33 (t, J = 7.36, 1.5 Hz, 1H), 7.28 (dd, J = 9.8, 2.4 Hz, 1H), 7.05 (d, J = 7.6 Hz, 1H), 7.02-6.99 (m, 1H). ¹³ C NMR (101 MHz, DMSO-d ₆ )δ163.3,160.6,158.2,140.0,136.5,136.4,129.6,129.5,128.8,123.5,123.0,122.5,122.4,120.1,110.9,109.6,109.3,98.5,98.2.HRMS(ESI,m/z):Calcd.for C ₁₅ H ₁₁ FN ₂ O[M+Na] ⁺ 277.0747,found:277.0754.

Embodiment 68:

Synthesis of 6-chloro-N-phenyl-1H-indole-3-carboxamide(C ₁₆ )

The preparation method is the same as that of Example 32, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 6-chloroindole-3-carboxylic acid, and compound C ₁₆ is finally obtained as a light yellow solid with a yield of 9%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.86 (s, 1H), 9.79 (s, 1H), 8.34 (s, 1H), 8.18 (d, J = 8.5 Hz, 1H), 7.76 (d, J = 8.0 Hz, 2H), 7.54 (d, J = 2.0 Hz, 1H), 7.33 (t, J = 7.7 Hz, 2H), 7.17 (dd, J = 8.6, 2.0 Hz, 1H), 7.04 (t, J = 7.4 Hz, 1H). ¹³ C NMR (101 MHz, DMSO-d ₆ )δ163.0,139.7,136.7,129.7,128.7,126.9,125.3,122.9,122.5,121.2,119.9,111.8,110.7. HRMS (ESI, m/z): Calcd. for C ₁₅ H ₁₁ ClN ₂ O[M+Na] ⁺ 293.0452, found: 293.0450.

Embodiment 69:

Synthesis of 6-cyano-N-phenyl-1H-indole-3-carboxamide(C ₁₇ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 6-cyanoindole-3-carboxylic acid, and compound C ₁₇ is finally obtained as a white solid with a yield of 6%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.26 (s, 1H), 9.88 (s, 1H), 8.56 (s, 1H), 8.34 (d, J = 8.4 Hz, 1H), 8.03 (s, 1H), 7.76 (d, J = 8.0 Hz, 2H), 7.50 (dd, J = 8.3, 1.5 Hz, 1H), 7.34 (t, J = 7.7 Hz, 2H), 7.06 (t, J = 7.4 Hz, 1H).

Embodiment 70:

Synthesis of 5-fluoro-N-phenyl-1H-indole-3-carboxamide(C ₁₈ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 5-fluoro-1H-indole-3-carboxylic acid, to finally obtain compound C ₁₈ as light green solid particles with a yield of 10%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.86 (s, 1H), 9.76 (s, 1H), 8.38 (s, 1H), 7.87 (dd, J = 10.2, 2.7 Hz, 1H), 7.77–7.75 (m, 2H), 7.49 (dd, J = 8.8, 4.6 Hz, 1H), 7.35–7.31 (m, 2H), 7.07–7.02 (m, 2H). ¹³ C NMR (101 MHz, DMSO-d ₆ ) δ 163.0, 156.8, 139.7, 132.9, 130.4, 128.6, 122.8, 119.8, 113.3, 113.2, 110.6, 105.9, 105.7.

Embodiment 71:

Synthesis of 6-bromo-N-phenyl-1H-indole-2-carboxamide(D ₀₁ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 6-bromoindole-2-carboxylic acid, to finally obtain compound D ₀₁ as a light yellow solid with a yield of 68%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.91 (s, 1H), 10.29 (s, 1H), 7.81–7.78 (m, 2H), 7.67 (d, J=8.6 Hz, 1H), 7.63 (s, 1H), 7.40 (s, 1H), 7.42–7.36 (m, 2H), 7.20 (dd, J=8.5, 1.8 Hz, 1H), 7.11 (t, J=7.4 Hz, 1H). ¹³ C NMR (101 MHz, DMSO-d ₆ ) δ 159.5, 138.9, 137.6, 132.4, 128.8, 126.1, 123.8, 123.8, 123.0, 120.3, 116.7, 114.9, 104.0.

Embodiment 72:

Synthesis of N-benzyl-6-bromo-1H-indole-2-carboxamide(D ₀₂ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 6-bromoindole-2-carboxylic acid, and aniline is replaced by benzylamine, to finally obtain compound D ₀₂ as a light yellow powder with a yield of 36%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.79 (s, 1H), 9.14 (t, J = 6.0 Hz, 1H), 7.61-7.59 (m, 2H), 7.34 (d, J = 4.4 Hz, 4H), 7.28-7.24 (m, 1H), 7.21 (s, 1H), 7.17 (dd, J = 8.6, 1.8 Hz, 1H), 4.52 (d, J = 6.1 Hz, 2H). ¹³ C NMR (101 MHz, DMSO-d ₆ )δ160.9,139.5,137.3,132.5,128.4,127.3,126.9,126.2,123.5,122.8,116.1,114.8,102.8,42.3. HRMS (ESI, m/z): Calcd. for C ₁₆ H ₁₃ BrN ₂ O[M+Na] ⁺ 351.0103, found: 351.0101.

Embodiment 73:

Synthesis of 6-bromo-N-(3-bromophenyl)-1H-indole-2-carboxamide(D ₀₃ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 6-bromoindole-2-carboxylic acid, and aniline is replaced by 3-bromoaniline, to finally obtain compound D ₀₃ as a white solid with a yield of 61%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.17 (s, 1H), 10.85 (s, 1H), 8.23 (t, J = 2.0 Hz, 1H), 7.94-7.92 (m, 1H), 7.64 (d, J = 8.9 Hz, 3H), 7.34-7.27 (m, 2H), 7.20 (dd, J = 8.5, 1.7 Hz, 1H). ¹³ C NMR (101 MHz, DMSO-d ₆ )δ159.6,140.7,137.6,132.1,130.7,126.2,126.0,123.8,123.1,122.4,121.5,118.9,116.8,114.9,105.2.HRMS(ESI,m/z):Calcd.for C ₁₅ H ₁₀ Br ₂ N ₂ O[M+Na] ⁺ 414.9052,found:414.9045.

Embodiment 74:

Synthesis of 6-bromo-N-(3-bromobenzyl)-1H-indole-2-carboxamide(D ₀₄ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 6-bromoindole-2-carboxylic acid, and aniline is replaced by 3-bromobenzylamine, to finally obtain compound D ₀₄ as a light yellow solid with a yield of 80%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.79 (s, 1H), 9.17 (t, J = 6.1 Hz, 1H), 7.62-7.59 (m, 2H), 7.53 (s, 1H), 7.47-7.44 (m, 1H), 7.36-7.29 (m, 2H), 7.20 (s, 1H), 7.17 (dd, J = 8.5, 1.8 Hz, 1H), 4.51 (d, J = 5.9 Hz, 2H). ¹³ C NMR (101 MHz, DMSO-d ₆ )δ160.9,142.4,137.3,132.3,130.6,130.0,129.8,126.4,126.1,123.6,122.8,121.7,116.2,114.8,102.9,41.7. HRMS (ESI, m/z): Calcd. for C ₁₆ H ₁₂ Br ₂ N ₂ O[M+Na] ⁺ 428.9208, found: 428.9204.

Embodiment 75:

Synthesis of 6-bromo-N-(3-fluorophenyl)-1H-indole-2-carboxamide(D ₀₅ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 6-bromoindole-2-carboxylic acid, and aniline is replaced by 3-fluoroaniline, to finally obtain compound D ₀₅ as a light yellow solid with a yield of 66%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.11 (s, 1H), 10.75 (s, 1H), 7.86 (dt, J = 11.8, 2.3 Hz, 1H), 7.70–7.63 (m, 3H), 7.58 (s, 1H), 7.43–7.37 (m, 1H), 7.20 (dd, J = 8.5, 1.8 Hz, 1H), 6.93 (td, J = 8.5, 2.6 Hz, 1H). ¹³ C NMR (101 MHz, DMSO-d ₆ )δ159.9,147.9,140.3,137.7,131.8,130.1,126.1,126.0,123.9,123.1,118.1,116.9,114.9,114.2,105.6.HRMS(ESI,m/z):Calcd.for C ₁₅ H ₁₀ BrFN ₂ O[M+Na] ⁺ 354.9853,found:354.9850.

Embodiment 76:

Synthesis of 6-bromo-N-(3-chlorophenyl)-1H-indole-2-carboxamide(D ₀₆ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 6-bromoindole-2-carboxylic acid, and aniline is replaced by 3-chloroaniline, to finally obtain compound D ₀₆ as a light yellow solid with a yield of 79%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.05 (s, 1H), 10.64 (s, 1H), 8.04 (t, J = 2.1 Hz, 1H), 7.80 (dd, J = 8.2, 2.0 Hz, 1H), 7.66 (d, J = 8.5 Hz, 1H), 7.64 (s, 1H), 7.54 (s, 1H), 7.40 (t, J = 8.1 Hz, 1H), 7.21 (dd, J = 8.5, 1.8 Hz, 1H), 7.16 (dd, J = 7.9, 2.1 Hz, 1H). ¹³ C NMR (101 MHz, DMSO-d ₆ )δ159.6,140.6,137.6,133.0,132.1,130.4,126.0,123.8,123.3,123.1,119.6,118.6,116.8,114.9,105.1. HRMS (ESI, m/z): Calcd. for C ₁₅ H ₁₀ BrClN ₂ O[M+H] ⁺ 348.9738, found: 348.9737.

Embodiment 77:

Synthesis of 6-bromo-N-(3-iodophenyl)-1H-indole-2-carboxamide(D ₀₇ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 6-bromoindole-2-carboxylic acid, and aniline is replaced by 3-iodoaniline, to finally obtain compound D ₀₇ as an off-white solid with a yield of 82%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.15 (s, 1H), 10.76 (s, 1H), 8.34 (t, J = 1.9 Hz, 1H), 7.95 (d, J = 8.2 Hz, 1H), 7.63 (t, J = 8.7 Hz, 3H), 7.45 (d, J = 7.8 Hz, 1H), 7.21–7.14 (m, 2H). ¹³ C NMR (101 MHz, DMSO-d ₆ ) δ 159.4, 140.5, 137.6, 132.1, 132.1, 130.8, 128.2, 126.0, 123.8, 123.0, 119.4, 116.8, 114.8, 105.0, 94.6. HRMS (ESI, m/z): Calcd. for C ₁₅ H ₁₀ BrIN ₂ O[M+Na] ⁺ 462.8913,found:462.8912.

Embodiment 78:

Synthesis of 6-bromo-N-(3-nitrophenyl)-1H-indole-2-carboxamide(D ₀₈ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 6-bromoindole-2-carboxylic acid, and aniline is replaced by 3-nitroaniline, to finally obtain compound D ₀₈ as a yellow powder with a yield of 78%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.19 (s, 1H), 11.21 (s, 1H), 8.93 (s, 1H), 8.38 (d, J = 8.2 Hz, 1H), 7.96 (dd, J = 8.2, 2.3 Hz, 1H), 7.70–7.68 (m, 2H), 7.66 (d, J = 2.4 Hz, 1H), 7.65 (s, 1H), 7.21 (dd, J = 8.4, 1.8 Hz, 1H). HRMS (ESI, m/z): Calcd. for C ₁₅ H ₁₀ BrN ₃ O ₃ [M+Na] ⁺ 381.9798, found: 381.9791.

Embodiment 79:

Synthesis of 6-bromo-N-(m-tolyl)-1H-indole-2-carboxamide (D ₀₉ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 6-bromoindole-2-carboxylic acid, and aniline is replaced by 3-methylaniline, to finally obtain compound D ₀₉ as a white needle-shaped solid with a yield of 39%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.88 (s, 1H), 10.21 (s, 1H), 7.66 (d, J = 8.5 Hz, 1H), 7.62 (d, J = 9.5 Hz, 3H), 7.45 (s, 1H), 7.25 (t, J = 7.7 Hz, 1H), 7.20 (dd, J = 8.5, 1.8 Hz, 1H), 6.93 (d, J = 7.5 Hz, 1H), 2.32 (s, 3H). ¹³ C NMR (101 MHz, DMSO-d ₆ )δ159.4,138.8,137.9,137.5,132.5,128.6,126.1,124.5,123.7,123.0,120.8,117.5,116.6,114.8,103.9,21.3. HRMS (ESI, m/z): Calcd. for C ₁₆ H ₁₃ BrN ₂ O[M+Na] ⁺ 351.0103, found: 351.0102.

Embodiment 80:

Synthesis of 6-bromo-N-(2-bromophenyl)-1H-indole-2-carboxamide(D ₁₀ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 6-bromoindole-2-carboxylic acid, and aniline is replaced by 2-bromoaniline, to finally obtain compound D ₁₀ as a light yellow solid with a yield of 28%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.95 (s, 1H), 10.15 (s, 1H), 7.74 (dd, J＝8.1, 1.4 Hz, 1H), 7.66 (d, J＝8.5 Hz, 1H), 7.62 (s, 1H), 7.58 (dd, J＝7.9, 1.6 Hz, 1H), 7.46 (td, J＝7.7, 1.4 Hz, 1H), 7.41 (d, J＝2.1 Hz, 1H), 7.25 (td, J＝7.7, 1.7 Hz, 1H), 7.21 (dd, J＝8.5, 1.8 Hz, 1H). HRMS (ESI, m/z): Calcd. for C ₁₅ H ₁₀ Br ₂ N ₂ O[MH] ⁺ 390.9087, found: 390.9076.

Embodiment 81:

Synthesis of 6-bromo-N-(4-bromophenyl)-1H-indole-2-carboxamide(D ₁₁ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 6-bromoindole-2-carboxylic acid, and aniline is replaced by 4-bromoaniline, to finally obtain compound D ₁₁ as a white solid with a yield of 81%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.94 (s, 1H), 10.41 (s, 1H), 7.79 (d, J = 8.8 Hz, 2H), 7.67 (d, J = 8.6 Hz, 1H), 7.63 (s, 1H), 7.57 (s, 1H), 7.55 (s, 1H), 7.46 (s, 1H), 7.21 (dd, J = 8.4, 1.8 Hz, 1H). ¹³ C NMR (101 MHz, DMSO-d ₆ ) δ 159.5, 138.3, 137.6, 132.1, 131.6, 126.0, 123.8, 123.1, 122.1, 116.8, 115.4, 114.9, 104.3. HRMS (ESI, m/z): Calcd. for C ₁₅ H ₁₀ Br ₂ N ₂ O[MH] ⁺ 390.9087,found:390.9092.

Embodiment 82:

Synthesis of 6-bromo-N-(3,5-dibromophenyl)-1H-indole-2-carboxamide(D ₁₂ )

The preparation method was the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid was replaced by 6-bromoindole-2-carboxylic acid, and aniline was replaced by 3,5-dibromoaniline, to finally obtain compound D ₁₂ as a white solid with a yield of 31%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.96 (s, 1H), 10.49 (s, 1H), 8.08 (d, J = 1.7 Hz, 2H), 7.69 (d, J = 8.6 Hz, 1H), 7.64 (s, 1H), 7.56 (t, J = 1.8 Hz, 1H), 7.45 (s, 1H), 7.22 (dd, J = 8.6, 1.8 Hz, 1H). ¹³ C NMR (101 MHz, DMSO-d ₆ ) δ 159.7, 141.6, 137.8, 131.5, 128.1, 125.9, 124.0, 123.2, 122.4, 121.2, 117.1, 114.9, 104.8. HRMS (ESI, m/z): Calcd. for C ₁₅ H ₉ Br ₃ N ₂ O[MH] ⁺ 468.8192,found:468.8192.

Embodiment 83:

Synthesis of 6-bromo-N-cyclohexyl-1H-indole-2-carboxamide (D ₁₃ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 6-bromoindole-2-carboxylic acid, and aniline is replaced by cyclohexylamine, to finally obtain compound D ₁₃ as a light yellow solid with a yield of 90%. ¹ H NMR (400 MHz, DMSO-d ₆ )δ11.70(s,1H),8.31(d,J＝8.0Hz,1H),7.59-7.57(m,2H),7.18(s,1H),7.15(dd,J＝8.6,1.7Hz,1H),3.78(s,1H),1.84(s,2H),1.73(s,2H),1.61(d,J＝12.7Hz,1H),1.36–1.26(m,4H),1.18–1.10(m,1H). ¹³ C NMR (101 MHz, DMSO-d ₆ )δ159.8,137.1,132.9,126.2,123.4,122.7,115.9,114.7,102.6,48.1,32.6,25.3,25.0.HRMS(ESI,m/z):Calcd.for C ₁₅ H ₁₇ BrN ₂ O[M+Na] ⁺ 343.0416,found:343.0415.

Embodiment 84:

Synthesis of 6-bromo-N-cyclopentyl-1H-indole-2-carboxamide (D ₁₄ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 6-bromoindole-2-carboxylic acid, and aniline is replaced by cyclopentylamine, to finally obtain compound D ₁₄ as a milky white solid with a yield of 70%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.70 (s, 1H), 8.38 (d, J = 7.5 Hz, 1H), 7.59-7.57 (m, 2H), 7.19 (s, 1H), 7.15 (dd, J = 8.5, 1.7 Hz, 1H), 4.29-4.20 (m, 1H), 1.94-1.86 (m, 2H), 1.74-1.66 (m, 2H), 1.59-1.49 (m, 4H). ¹³ C NMR (101 MHz, DMSO-d ₆ )δ160.4,137.1,132.9,126.2,123.4,122.7,115.9,114.7,102.7,50.7,32.2,23.7.HRMS(ESI,m/z):Calcd.for C ₁₄ H ₁₅ BrN ₂ O[M+Na] ⁺ 329.0260,found:329.0258.

Embodiment 85:

Synthesis of 6-bromo-N-(thiazol-2-yl)-1H-indole-2-carboxamide (D ₁₅ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 6-bromoindole-2-carboxylic acid, and aniline is replaced by 2-aminothiazole, to finally obtain compound D ₁₅ as a light yellow solid with a yield of 13%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.81 (s, 1H), 12.07 (s, 1H), 7.67–7.64 (m, 3H), 7.57 (d, J=3.5 Hz, 1H), 7.29 (d, J=3.5 Hz, 1H), 7.21 (dd, J=8.5, 1.8 Hz, 1H).

Embodiment 86:

Synthesis of 6-bromo-N-(pyridin-2-yl)-1H-indole-2-carboxamide(D ₁₆ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 6-bromoindole-2-carboxylic acid, and aniline is replaced by 2-aminopyridine, to finally obtain compound D ₁₆ as a white solid with a yield of 9%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.05 (s, 1H), 10.97 (s, 1H), 8.41 (d, J = 4.9 Hz, 1H), 8.23 (d, J = 8.3 Hz, 1H), 7.86 (t, J = 7.9 Hz, 1H), 7.66–7.62 (m, 3H), 7.21–7.16 (m, 2H). ¹³ C NMR (101 MHz, DMSO-d ₆ ) δ 159.8, 152.0, 148.0, 138.3, 137.7, 131.7, 126.1, 124.0, 123.1, 119.8, 116.9, 114.9, 114.7, 105.3.

Embodiment 87:

Synthesis of 6-bromo-N-(pyridin-3-yl)-1H-indole-2-carboxamide (D ₁₇ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 6-bromoindole-2-carboxylic acid, and aniline is replaced by 3-aminopyridine, to finally obtain compound D ₁₇ as a white solid with a yield of 75%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.00 (s, 1H), 10.52 (s, 1H), 8.97 (s, 1H), 8.33 (d, J = 4.6 Hz, 1H), 8.21 (d, J = 8.3 Hz, 1H), 7.69 (d, J = 8.5 Hz, 1H), 7.64 (s, 1H), 7.48 (s, 1H), 7.42 (dd, J = 8.3, 4.7 Hz, 1H), 7.22 (d, J = 7.9 Hz, 1H). ¹³ C NMR (101 MHz, DMSO-d ₆ )δ159.9,144.7,141.8,137.7,135.6,131.9,127.3,126.1,123.9,123.8,123.2,116.9,114.9,104.5. HRMS (ESI, m/z): Calcd. for C ₁₄ H ₁₀ BrN ₃ O[M+H] ⁺ 316.0080, found: 316.0072.

Embodiment 88:

Synthesis of N-(3-bromophenyl)-1H-indole-2-carboxamide(D ₁₈ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by indole-2-carboxylic acid, and aniline is replaced by 3-bromoaniline, to finally obtain compound D ₁₈ as a white block solid with a yield of 23%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.79 (s, 1H), 10.35 (s, 1H), 8.13 (t, J = 2.0 Hz, 1H), 7.82-7.79 (m, 1H), 7.69 (dd, J = 8.0, 1.0 Hz, 1H), 7.48 (dd, J = 8.2, 1.0 Hz, 1H), 7.44 (dd, J = 2.2, 1.0 Hz, 1H), 7.34 (t, J = 8.0 Hz, 1H), 7.31-7.28 (m, 1H), 7.26-7.22 (m, 1H), 7.10-7.06 (m, 1H). ¹³ C NMR (101 MHz, DMSO-d ₆ )δ159.9,140.7,137.0,131.1,130.8,127.0,126.1,124.1,122.3,121.9,121.6,120.1,118.8,112.5,104.4.

Embodiment 89:

Synthesis of 6-fluoro-N-phenyl-1H-indole-2-carboxamide(D ₁₉ )

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 6-fluoroindole-2-carboxylic acid, and compound _D19 is finally obtained as a light yellow solid with a yield of 79%. 1H NMR (400MHz, DMSO-d6) δ11.87 (s, 1H), 10.24 (s, 1H), 7.81 (d, J = 7.2 Hz, 1H), 7.71 (dd, J = 8.8, 5.5 Hz, 1H), 7.47 (d, J = 2.1 Hz, 1H), 7.39-7.35 (m, 2H), 7.19 (dd, J = 10.0, 2.4 Hz, 1H), 7.13-7.08 (m, 1H), 6.96 (td, J = 9.9, 18.5, 2.4 Hz, 1H).13C NMR (101 MHz, DMSO-d6) δ 161.4, 159.5, 159.0, 138.9, 136.9, 136.8, 132.4, 132.4, 128.8, 124.0, 123.6, 123.4, 123.3, 120.2, 109.3, 109.0, 104.1, 98.1, 97.8. HRMS (ESI, m/z): Calcd. for C15H11FN2O [M+Na] + 277.0747, found: 277.0748.

Embodiment 90:

Synthesis of 5-methoxy-N-phenyl-1H-indole-2-carboxamide (D ₂₀ ):

The preparation method is the same as that of Example 53, except that 6-bromo-1H-indole-3-carboxylic acid is replaced by 5-methoxyindole-2-carboxylic acid, to finally obtain compound D ₂₀ as a brown solid with a yield of 65%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.63 (s, 1H), 10.18 (s, 1H), 7.81 (d, J＝7.4 Hz, 2H), 7.39–7.35 (m, 4H), 7.14–7.08 (m, 2H), 6.88 (dd, J＝8.8, 2.5 Hz, 1H), 3.78 (s, 3H). ¹³ C NMR (101 MHz, DMSO-d ₆ ) δ 159.8, 153.9, 139.1, 132.2, 131.9, 128.8, 127.4, 123.6, 120.3, 115.2, 113.3, 103.7, 102.1, 55.3. HRMS (ESI, m/z): Calcd. for C ₁₆ H ₁₄ N ₂ O ₂ [M+Na] ⁺ 289.0949, found: 289.0947.

Bioactivity screening

GloSensor ^TM cAMP Assay measures cAMP levels:

cAMP is a key signaling molecule for many G protein-coupled receptors. The accumulation level of cAMP is mainly tested using GloSensor, a bioluminescence-based biosensor that can directly detect intracellular cAMP (Promega). The principle is to insert a cAMP binding domain into the N-terminus and C-terminus of firefly luciferase through genetic engineering technology to make the enzyme in an inactive state. When cAMP binds to the cAMP binding domain, the enzyme is activated, thereby oxidizing the substrate luciferin to produce bioluminescence. The cAMP accumulation experiment is used to test the functional activity of the target compound on β2AR and to clarify whether the new compound is a negative allosteric modulator (NAM) of _β2AR . Briefly described as follows, HEK 293T cells were seeded in 6-well plates, with 4×10 ⁵ cells per well. The next day, β2AR and pGloSensor-22F cAMP plasmids were transfected into HEK 293T cells simultaneously using FuGene transfection reagent (Promega). After 48 hours, the transfected cells were washed with _CO2- independent medium and incubated with a 2% v/v GloSensor cAMP reagent stock solution (dissolved in _CO2- independent medium containing 10% FBS). After incubation at 37°C for 1 hour and then at room temperature for 1 hour, the bioluminescent signal was detected by a multifunctional microplate reader until a steady-state baseline signal was obtained. Then, different concentration gradients of the new derivative and the control compound Cmpd-15 were added to the cells, and the positive control ISO (final concentration 1nM-100μM) was added after incubation at 37°C for 30 minutes. The changes in bioluminescence were read with a microplate reader.

The GloSensor cAMP accumulation test was used to compare the allosteric antagonistic activity of the new pyrazole derivatives (final concentration of 50 μM) with the lead compound Cmpd-15, using different concentration gradients of isoproterenol (ISO) as the positive control (final concentration of 1nM-100μM) and compound Cmpd-15 (final concentration of 50μM) as the reference control. The test results in Table 2 show that most compounds have allosteric antagonistic activity against β2AR, among which compound B8 has the highest activity, which is 4.17 times that of Cmpd-15.

Table 2-1. Comparison of allosteric activity of benzimidazole amide compounds relative to Cmpd-15

Table 2-2. Comparison of allosteric activity of benzopyrazole amide compounds relative to Cmpd-15

Table 2-3. Comparison of the allosteric activity of indoleamide compounds relative to Cmpd-15


Note: a value represents the blocking activity relative to Cmpd-15; “-” means the compound has no allosteric antagonistic activity

Allosteric antagonism mechanism studies

In addition, the cAMP accumulation test was used to further study the allosteric antagonism mechanism, and the target compounds were tested to determine whether they could allosterically regulate the functional activity of the β ₂ -AR endogenous ligand ISO to determine whether they were β ₂ -AR negative allosteric modulators (NAMs). Briefly, the new compounds (1nM-100μM) with multiple concentrations were first added to the cells, and after incubation at 37°C for 30 minutes, different concentration gradients of positive control ISO (final concentration 1nM-100μM) were added to the cells to test whether the change in bioluminescence at this time showed a concentration-dependent limited downward trend. The specific experimental steps were the same as the above Glosensor cAMP accumulation test. The experimental results showed that the target compounds A08, A26, B8, C01, C15, and C17 could negatively allosterically regulate the functional activity of the β ₂ -AR endogenous ligand ISO.

Claims (7)

A benzyl azoheterocyclic compound, characterized in that the structural formula of the benzyl azoheterocyclic compound is as follows:
X＝N,CH; Y＝N,CH; R ₁ =H, methyl, methoxy, phenyl, F, Cl, Br, I, CN, NO ₂ , NH ₂ , OH, CF ₃ ; R ₂ =H, methyl, phenyl, benzyl; R ₃ =H, alkyl; R ₄ =H, alkyl, cycloalkyl, phenyl, substituted phenyl, substituted benzyl. The benzazaheterocyclic compound according to claim 1 is characterized in that the benzazaheterocyclic compound is a benzimidazole amide compound, and its general structural formula is shown in Formula I:
Wherein R ₁ is H, methyl, methoxy, phenyl, F, Cl, Br, I, CN, NO ₂ , NH ₂ , OH, CF ₃ ; R ₃ is H, alkyl; R ₄ is H, phenyl, bromophenyl, fluorophenyl. The benzazaheterocyclic compound according to claim 1 is characterized in that the benzazaheterocyclic compound is a benzopyrazole amide compound, and its general structural formula is shown in Formula II:
Wherein R ₁ is H, methyl, methoxy, phenyl, F, Cl, Br, I, CN, NO ₂ , NH ₂ , OH, CF ₃ ; R ₂ is H, methyl, methoxy, phenyl, benzyl; R ₃ is H, alkyl; R ₄ is H, phenyl, bromophenyl, fluorophenyl. The benzazaheterocyclic compound according to claim 1, characterized in that the benzazaheterocyclic compound is an indoleamide compound, and its general structural formula is shown in Formula III:
Wherein R ₁ is H, methyl, methoxy, phenyl, F, Cl, Br, I, CN, NO ₂ , NH ₂ , OH, CF ₃ ; R ₃ is H, alkyl; R ₄ is H, phenyl, bromophenyl, fluorophenyl. The benzazazepine heterocyclic compound according to claim 1, characterized in that the _{benzazazepine} heterocyclic compound is an allosteric modulator of β ₂ -adrenergic receptor (β ₂ -AR). A method for synthesizing a benzazazepine compound according to any one of claims 1 to 5, characterized in that a simple amide coupling reaction occurs between 3-carboxylic acid benzazazepine and various amines, firstly dissolving 3-carboxylic acid benzazazepine and an activator in a solvent, adding different types of amines under an ice bath, then adding an acid binding agent and an amide coupling agent and continuing to stir to room temperature; the activator is 1-hydroxy-7-azabenzotriazole (HOAT), the acid binding agent is N-methylmorpholine (NMM), and the amide coupling agent is 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI); the molar ratio of 3-carboxylic acid benzazazepine: HOAT: amine: NMM: EDCI is 1:1.2-1.5:1-2:0.6-0.75:1.2-1.5. An application of the benzazazepine heterocyclic compound according to claim 1, characterized in that the benzazazepine heterocyclic compound is used in the preparation of β ₂ -adrenaline receptor allosteric antagonist drugs.