Indazole-based ALK5 inhibitors, their preparation methods and uses

Abstract

The present invention belongs to the field of medicinal chemistry, and discloses indazole compounds with the structure shown in formula III or their stereoisomers, pharmaceutically acceptable salts, prodrugs or solvates, where Ar is a substituted aromatic ring or a six-membered heteroaromatic ring; R is selected from Y being a heteroatom; R 1 and R 2 are each independently selected from hydrogen, fluorine, hydroxyl, aldehyde group, carbonyl group, carboxyl group, cyano group, formamido group, nitro group, C1-C6 alkyl group, C3-C6 cycloalkyl group, C3-C8 cycloheteroalkyl group containing 1 oxygen atom, C6-C 10 aryl group. The indole compounds of the present invention have significant inhibitory activity against ALK5, and the inhibition rate can reach up to 68% - 70% at a concentration of 50 nM, which is comparable to that of the lead compound, and can be used to block the TGF-β / SMAD signaling pathway. The present invention discloses the use of the indazole compounds or their stereoisomers, pharmaceutically acceptable salts, prodrugs or solvates in the preparation of ALK5 inhibitors.

Description

Technical Field

[0001] The present invention belongs to the field of medicinal chemistry, and relates to 3-aryl-5-aminoindazole compounds, their preparation methods and uses. Specifically, it relates to 3-aryl-5-aminoindazole compounds, their preparation methods, pharmaceutical compositions containing these compounds or their pharmaceutically acceptable salts, and their uses in blocking the ALK5 signaling pathway. Background Art

[0002] TGF-β (transforming growth factor-β) is an important regulator involved in the activities of numerous signaling pathways in major cellular processes such as cell proliferation, migration, survival, and differentiation. To date, five different TGF-β (TGF-β1-5) family isomers have been reported. Among them, TGF-β1 is mainly expressed in mammalian endothelial cells, hematopoietic cells, and connective tissue cells, with a proportion of over 90%. It is a key mediator in the pathogenesis of liver fibrosis.

[0003] ALK5 is an important type I receptor of the TGF-β1 family. TGF-β signals through two related transmembrane type I and type II serine / threonine kinase receptors. After active TGF-β binds to TGF-βII, the type I receptor (activin receptor-like kinase 5, ALK5) is phosphorylated and generates a binding site for Smad2 / Smad3 proteins, and Smad2 / Smad3 proteins are further phosphorylated. The phosphorylated Smad2 / Smad3 proteins form a heteromeric complex with Smad4, and Smad4 translocates into the nucleus, assembles with specific DNA-binding cofactors and co-regulators, and binds to the promoters of TGF-β target genes involved in cell differentiation, proliferation, apoptosis, migration, and extracellular matrix production. ALK5 plays an important role in TGF-β signal transduction. Therefore, the research on ALK5 inhibitors has attracted the attention of many drug researchers. Most of the reported ALK-5 inhibitors are small molecule inhibitors. Representative ALK5 small molecule inhibitors in clinical phase I or II studies are EW-7197, LY2157299, GFH018, SH3051, and YL-13027. Most of these compounds are used in the research for treating tumors. Among the reported small molecule inhibitors or biologics tested for anti-liver fibrosis, there is currently no inhibitor entering clinical research for anti-liver fibrosis. Summary of the Invention

[0004] The object of the invention is that in view of the current lack of small molecule drugs for effectively treating liver fibrosis diseases, the present invention takes ALK5 as a target, develops small molecule compounds with 3-aryl-5-aminoindazole as the core, and obtains good ALK5 inhibitory activity and has a therapeutic effect on liver fibrosis.

[0005] The object of the present invention is achieved by the following technical solutions:

[0006] An indazole compound having a structure as shown in Formula III, or a stereoisomer, pharmaceutically acceptable salt, prodrug or solvate thereof:

[0007]

[0008] Wherein:

[0009] Ar is a substituted aromatic ring or a six-membered heteroaromatic ring;

[0010] R is selected from

[0011] Y is a heteroatom;

[0012] R 1 and R 2 are each independently selected from hydrogen, fluorine, hydroxyl, aldehyde group (-CHO), carbonyl group, carboxyl group, cyano group (-CN), formamido group -CONH2, nitro group, C1-C6 alkyl group, C3-C6 cycloalkyl group, C3-C8 cycloheteroalkyl group containing one oxygen atom, C6-C 10 aryl group.

[0013] Preferably, Ar is selected from 3-pyridyl group and 3-nitrophenyl group;

[0014] Y is selected from oxygen atom and sulfur atom; R 1 is selected from hydrogen, fluorine, cyano group, formamido group, nitro group;

[0015] R 2 is selected from tetrahydropyran-4-yl group, cyclohexyl group, phenyl group.

[0016] More preferably, Ar is selected from 3-pyridyl group and 3-nitrophenyl group;

[0017] Y is oxygen atom and sulfur atom; R 1 is selected from -H, o-F, m-F, p-F, m-CN, p-CN, m-CONH2, p-CONH2;

[0018] R 2 is selected from

[0019] As a further technical solution of the present invention, an indazole compound having a structure as shown in Formula I, or a stereoisomer, pharmaceutically acceptable salt, prodrug or solvate thereof:

[0020]

[0021] Wherein: Ar, Y, R 1 are as described above, and are the same as Ar, Y, R of the indazole compound represented by Formula III 1The same.

[0022] As a further technical solution of the present invention, an indazole compound having a structure as shown in Formula II or a stereoisomer, pharmaceutically acceptable salt, prodrug or solvate thereof:

[0023]

[0024] Wherein: Ar, R 2 As described above, the Ar and R of the indazole compound represented by Formula III 2 The same.

[0025] Specifically, an indole compound having the following structure or a pharmaceutically acceptable salt thereof:

[0026]

[0027]

[0028] As a preferred technical solution of the present invention, an indazole compound having a structure as shown in Formula III or a stereoisomer, pharmaceutically acceptable salt, prodrug or solvate thereof:

[0029]

[0030] Wherein:

[0031] Ar is selected from 3-pyridyl, 3-nitrophenyl;

[0032] R is selected from

[0033] Y is an oxygen atom; R 1 Is selected from o-F, m-CONH2;

[0034] R 2 Is selected from

[0035] Excluding: Ar is selected from 3-nitrophenyl, and R is selected from R 2 Is selected from

[0036] An indazole compound having the following structure or a stereoisomer, pharmaceutically acceptable salt, prodrug or solvate thereof:

[0037]

[0038] Wherein:

[0039] Ar is selected from 3-pyridyl;

[0040] R is selected from

[0041] Y is an oxygen atom; R 1 is selected from o-F;

[0042] R 2 is selected from

[0043] The pharmaceutically acceptable salt is an acid addition salt formed by an indazole compound and the following acids: hydrogen chloride, hydrogen bromide, sulfuric acid, carbonic acid, oxalic acid, citric acid, succinic acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or ferulic acid.

[0044] The solvate may be a hydrate of the indazole compound.

[0045] Another object of the present invention is to provide the use of the indazole compound or its stereoisomer, pharmaceutically acceptable salt, prodrug or solvate in the preparation of an ALK5 inhibitor.

[0046] Another object of the present invention is to provide the use of the indazole compound or its stereoisomer, pharmaceutically acceptable salt, prodrug or solvate in the preparation of a drug for treating liver fibrosis.

[0047] Another object of the present invention is to provide a pharmaceutical composition, which uses the indazole compound or its stereoisomer, pharmaceutically acceptable salt, prodrug or solvate as an active ingredient.

[0048] Advantages of the present invention:

[0049] The indole compound of the present invention has the characteristics of novel skeleton, strong plasticity and great potential for future modification. It has significant inhibitory activity against ALK5, and the inhibition rate can reach up to 68% - 70% at a concentration of 50 nM, which is comparable to the lead compound. It can be used to block the TGF-β / SMAD signaling pathway and can be used to develop anti-liver fibrosis drugs with strong activity. Specific implementation methods

[0050] Example 1

[0051] N-(3-(Pyridin-3-yl)-1H-indazol-5-yl)benzamide (I-1a)

[0052]

[0053] Step (1): Synthesis of 3-bromo-5-nitro-1H-indazole (Compound 2)

[0054] 5-Nitro-1H-indazole (Compound 1, 5 g, 30.65 mmol) was added to 10 mL of DMF. After potassium hydroxide (5.16 g, 91.95 mmol) was crushed, it was added to the above mixed solution. The mixture was sonicated until part of the potassium hydroxide was dissolved, and then stirred for 10 min to fully mix Compound 1 with KOH. N-Bromosuccinimide (8.18 g, 45.97 mmol) was added in batches. The mixture was heated and stirred at 80 °C, and monitored by TLC until the raw material spot completely reacted. Heating was stopped. After the reaction solution cooled to room temperature, 20 mL of ethyl acetate was added to the reaction solution, and it was washed successively with saturated NaHCO3 solution (30 mL × 3) and saturated NaCl solution (30 mL × 6). The obtained organic phase was dried over anhydrous Na2SO4, filtered, and the organic phase was concentrated under reduced pressure. The obtained residue was triturated and subjected to silica gel column chromatography (eluent: PE:EA = 8:1 V / V) to obtain Compound 2 (yellow solid, 6.6 g, 27.28 mmol, yield 89%).

[0055] 1 H NMR (300 MHz, DMSO-d6) δ 14.11 (s, 1H), 8.48 (d, J = 1.7 Hz, 1H), 8.26 (dd, J = 9.2, 2.2 Hz, 1H), 7.78 (d, J = 9.8 Hz, 1H).

[0056] Step (2): Synthesis of 3-bromo-5-nitro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazole (Compound 3)

[0057] Compound 2 (5.94 g, 24.54 mmol) was dissolved in 40 mL of dichloromethane. A catalytic amount of tetrabutylammonium bromide (0.08 g, 0.25 mmol) and 50% (wt) aqueous potassium hydroxide solution (40 mL) were added successively. During this process, the color of the reaction solution gradually became darker. The reaction solution was transferred to an ice bath. After the reaction system cooled to 0 °C, 2-(trimethylsilyl)ethoxymethyl chloride (4.89 mL, 27.00 mmol) was slowly added dropwise. The reaction was monitored by TLC until the raw material spot disappeared. The ice bath was removed and the reaction was stopped. The reaction solution was concentrated, 20 mL of ethyl acetate was added, and it was washed with water (30 mL × 3) respectively. The organic phase was washed with saturated NaCl solution (30 mL × 3), dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure. The obtained residue was triturated and subjected to silica gel column chromatography (PE:EA = 12:1 V / V) to obtain Compound 3 (yellow solid, 7.86 g, 21.10 mmol, yield 86%).

[0058] 11H NMR (300 MHz, Chloroform-d) δ 8.63 (d, J = 1.5 Hz, 1H), 8.36 (dd, J = 9.2, 2.1 Hz, 1H), 7.66 (d, J = 9.9 Hz, 1H), 5.73 (s, 2H), 3.59 (t, 2H), 0.89 (t, 2H), -0.06 (s, 9H).

[0059] Step (3): Synthesis of 3-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-amine (Compound 4)

[0060] Dissolve Compound 3 (7.2 g, 19.34 mmol) in 30 mL of a mixed solvent (ethanol:water = 10:1 V / V). Add iron powder (5.40 g, 96.70 mmol) and anhydrous ammonium chloride (3.10 g, 58.02 mmol) in sequence. After stirring at room temperature for 10 min to activate the iron powder, then heat and stir at 80 °C for 1 h. Monitor by TLC and stop heating when the starting material spot disappears. Cool to room temperature. Filter the reaction solution through diatomaceous earth, concentrate under reduced pressure to remove ethanol, add 20 mL of ethyl acetate. Wash the organic phase successively with saturated NaHCO3 solution (30 mL × 3), saturated NaCl solution (30 mL × 3), dry over anhydrous Na2SO4, filter, concentrate under reduced pressure. Grind the obtained residue and perform silica gel column chromatography (PE:EA = 12:1 V / V) to obtain Compound 4 (brownish-yellow oily substance, 7.86 g, 21.10 mmol, yield 94%).

[0061] 1 1H NMR (300 MHz, DMSO-d6) δ 7.47 (d, J = 8.9 Hz, 1H), 6.92 (dd, J = 8.9, 2.0 Hz, 1H), 6.56 (d, J = 2.0 Hz, 1H), 5.59 (s, 2H), 5.14 (s, 2H), 3.48 (t, J = 8.0 Hz, 2H), 0.78 (t, J = 7.9 Hz, 2H), -0.11 (s, 9H); 13 13C NMR (75 MHz, DMSO-d6) δ 140.23, 127.60, 124.32, 123.78, 121.45, 114.15, 112.76, 77.74, 66.45, 17.57.

[0062] Step (4): Synthesis of 3-(3-pyridyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-amine (Compound 5)

[0063] Compound 4 (2.20 g, 6.43 mmol), 3-Pyridylboronic acid (0.87 g, 7.07 mmol), tetrakis(triphenylphosphine)palladium(0) (0.74 g, 0.64 mmol), and potassium carbonate (2.66 g, 19.28 mmol) were placed in a three-necked flask. After purging with nitrogen three times, 30 mL of a mixed solvent (dioxane:H2O = 7:1 V / V) was added. After stirring at room temperature for 10 min to activate the catalyst, the mixture was heated and stirred at 100 °C. The reaction was monitored by TLC until completion, then heating was stopped and the mixture was cooled to room temperature. The reaction solution was filtered through diatomaceous earth, and the filtrate was extracted with dichloromethane (30 mL × 3). The organic phase was washed successively with saturated NaHCO3 solution (30 mL × 3), saturated NaCl solution (30 mL × 3), dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure. The resulting residue was triturated and subjected to silica gel column chromatography (eluent: PE:EA = 3:1 V / V) to obtain Compound 5 (pale yellow oil, 1.44 g, 4.24 mmol, yield 66%).

[0064] 1 H NMR (300 MHz, DMSO-d6) δ 9.10 (s, 1H), 8.58 (d, J = 4.8 Hz, 1H), 8.24 (d, J = 7.9 Hz, 1H), 7.65 - 7.43 (m, 2H), 7.15 (s, 1H), 6.92 (d, J = 8.8 Hz, 1H), 5.71 (s, 2H), 5.06 (s, 2H), 3.56 (t, J = 7.9 Hz, 2H), 0.81 (t, J = 7.9 Hz, 2H), -0.11 (s, 9H).

[0065] Step (5): Synthesis of N-((3-Pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)benzamide (Compound 6a)

[0066] Benzoic acid (0.063 g, 0.52 mmol) and a catalytic amount of 4-dimethylaminopyridine (0.006 g, 0.051 mmol) were added to dichloromethane (3 mL). Carbodiimide hydrochloride (0.20 g, 0.52 mmol) was added in portions, and the mixture was stirred at room temperature for 10 min. Compound 5 (0.17 g, 0.51 mmol) was added, and the reaction was carried out at room temperature. The reaction was monitored by TLC until completion, then the reaction was stopped. The reaction solution was washed successively with saturated NaHCO3 solution (30 mL × 3), saturated NaCl solution (30 mL × 3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain Compound 6a (reddish-brown oil), which was used directly in the next step without purification.

[0067] Step (6): Synthesis of N-(3-(pyridin-3-yl)-1H-indazol-5-yl)benzamide (Compound I-1a)

[0068] Dissolve Compound 6a (0.11 g, 0.247 mmol) in tetrahydrofuran (3 mL). While stirring, add ethylenediamine (0.05 mL, 0.742 mmol) and tetrabutylammonium fluoride solution (1 M, 1.24 mL, 1.29 mmol, prepared from THF) in sequence. Reflux the reaction until TLC monitoring shows that the reaction is complete. Stop heating and cool to room temperature. Distill off part of the solvent under reduced pressure, add 10 mL of ethyl acetate, wash with saturated NaHCO3 solution (15 mL × 3) in sequence, wash with saturated NaCl solution (15 mL × 3), dry over anhydrous Na2SO4, filter, concentrate under reduced pressure to obtain a pale yellow crude product. After dissolving the crude product in a small amount of tetrahydrofuran, add a large amount of petroleum ether. White solid precipitates. Filter by suction, wash the filter cake with a small amount of ethyl acetate, and dry to obtain Compound I-1a (white solid, 0.036 mg, yield 46%).

[0069] Mp: 250 - 252 °C; 1 1H NMR (300 MHz, DMSO-d6) δ 13.43 (s, 1H), 10.36 (s, 1H), 9.17 (s, 1H), 8.62 (d, J = 6.1 Hz, 2H), 8.32 (d, J = 8.0 Hz, 1H), 8.01 (d, J = 7.2 Hz, 2H), 7.85 (d, J = 9.0 Hz, 1H), 7.69 - 7.48 (m, 5H); 13 13C NMR (75 MHz, DMSO-d6) δ 165.82, 148.96, 147.60, 140.64, 139.01, 135.31, 134.06, 133.80, 131.90, 130.11, 128.78, 127.95, 124.49, 121.88, 120.27, 111.15, 110.86; HRMS (+ESI) m / z calcd for C 19 H 14 N4O [M + H] + 314.1168, found 314.1168.

[0070] Example 2

[0071] 2-Fluoro-N-(3-(pyridin-3-yl)-1H-indazol-5-yl)benzamide (Compound I-1b)

[0072] Step (1): Synthesis of 2-fluoro-N-(3-(pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)benzamide (6b)

[0073] Referring to the method described in step (5) of Reference Example 1, replace benzoic acid with o-fluorobenzoic acid in equimolar amounts to obtain compound 6b (reddish-brown oil).

[0074] Step (2): Synthesis of 2-fluoro-N-(3-(pyridin-3-yl)-1H-indazol-5-yl)benzamide (Compound I-1b)

[0075] Referring to the method described in step (6) of Reference Example 1, replace compound 6a with compound 6b in equimolar amounts to obtain compound I-1b, a white solid, with a yield of 40%.

[0076] Mp: 254 - 256 °C; 1 H NMR (300 MHz, DMSO-d6) δ 13.52 (s, 1H), 10.54 (s, 1H), 9.16 (s, 1H), 8.62 (s, 2H), 8.30 (d, J = 7.9 Hz, 1H), 7.76 - 7.53 (m, 5H), 7.36 (q, J = 8.3, 7.3 Hz, 2H); 13 C NMR (75 MHz, DMSO-d6) δ 163.25, 161.08, 157.78, 149.14, 147.78, 140.89, 139.21, 134.27, 133.71, 133.09, 132.98, 130.47, 130.22, 125.62, 125.42, 125.13, 124.66, 121.43, 120.43, 116.85, 116.57, 111.57, 110.38; HRMS (+ESI) m / z calcd for C 19 H 13 ON4F [M + H] + 332.1073, found 332.1078. Example 3

[0077] 3-fluoro-N-(3-(pyridin-3-yl)-1H-indazol-5-yl)benzamide (Compound I-1c)

[0078] Step (1): Synthesis of 3-fluoro-N-(3-(pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)benzamide (Compound 6c)

[0079] Referring to the method described in step (5) of Reference Example 1, replace benzoic acid with m-fluorobenzoic acid in equimolar amounts to obtain a reddish-brown oil. Grind it into sand and perform silica gel column chromatography (eluent: DCM:MeOH = 200:1 V / V) to purify and obtain Compound 6c, a pale yellow solid, with a yield of 70%.

[0080] 1 H NMR (300 MHz, DMSO-d6) δ 10.46 (s, 1H), 9.16 (d, J = 1.7 Hz, 1H), 8.66 (dd, J = 4.7, 1.7 Hz, 1H), 8.60 (s, 1H), 8.30 (dt, J = 8.1, 2.0 Hz, 1H), 7.94 - 7.79 (m, 4H), 7.62 (dt, J = 8.4, 5.7 Hz, 2H), 7.46 (td, J = 8.2, 2.3 Hz, 1H), 5.84 (s, 2H), 3.61 (t, J = 7.9 Hz, 2H), 0.83 (t, J = 7.9 Hz, 2H), -0.11 (s, 9H); 13 C NMR (75 MHz, DMSO-d6) δ 163.64, 163.10, 159.86, 148.64, 146.94, 140.16, 137.95, 136.69, 136.59, 133.51, 133.39, 130.21, 130.10, 128.46, 123.73, 123.37, 123.34, 121.42, 120.75, 118.19, 117.91, 114.12, 113.82, 110.51, 110.21, 76.48, 65.33, 16.62, -1.91.

[0081] Step (2): Synthesis of 3-fluoro-N-(3-(pyridin-3-yl)-1H-indazol-5-yl)benzamide (Compound I-1c)

[0082] Referring to the method described in step (6) of Reference Example 1, replace Compound 6a with Compound 6c in equimolar amounts to obtain Compound I-1c, a white solid, with a yield of 42%.

[0083] Mp: 262 - 264 °C; 1 H NMR (300 MHz, DMSO-d6) δ 13.46 (s, 1H), 10.43 (s, 1H), 9.17 (s, 1H), 8.68 - 8.55 (m, 2H), 8.31 (dt, J = 8.0, 2.0 Hz, 1H), 7.93 - 7.76 (m, 3H), 7.69 - 7.53 (m, 3H), 7.52 - 7.39 (m, 1H); 1313C NMR(75MHz, DMSO-d6) δ 165.13, 164.64, 161.40, 149.67, 148.31, 141.40, 139.84, 138.25, 134.79, 134.19, 131.71, 131.60, 130.79, 125.20, 124.90, 122.63, 120.96, 119.64, 119.36, 115.67, 115.36, 111.84; HRMS(+ESI) m / z calcd for C 19 H 13 ON4F [M + H] + 332.1073, found 332.1078.

[0084] Example 4

[0085] 4-Fluoro-N-(3-(pyridin-3-yl)-1H-indazol-5-yl)benzamide (Compound I-1d)

[0086] Step (1): Synthesis of 4-fluoro-N-(3-(pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)benzamide (Compound 6d)

[0087] Referring to the method described in Step (5) of Reference Example 1, replace benzoic acid with p-fluorobenzoic acid in equimolar amounts to obtain a reddish-brown oil. Perform silica gel column chromatography (eluent: DCM:MeOH = 200:1 V / V) to purify and obtain Compound 6d as a white solid with a yield of 72%.

[0088] 1 1H NMR(300MHz, DMSO-d6) δ 10.44(s, 1H), 9.20 - 9.10(m, 1H), 8.66(dd, J = 4.8, 1.6Hz, 1H), 8.59(s, 1H), 8.30(dt, J = 8.1, 1.9Hz, 1H), 8.09(dd, J = 8.6, 5.6Hz, 2H), 7.95 - 7.81(m, 2H), 7.61(dd, J = 8.1, 4.9Hz, 1H), 7.40(t, J = 8.8Hz, 2H), 5.84(s, 2H), 3.60(t, J = 7.9Hz, 2H), 0.83(t, J = 8.0Hz, 2H), -0.11(s, 10H).; 1313C NMR (75 MHz, DMSO-d6) δ 163.77, 161.78, 148.44, 146.74, 139.94, 137.70, 133.39, 133.32, 130.56, 129.73, 129.61, 128.29, 123.54, 121.27, 120.58, 114.86, 114.58, 110.21, 109.99, 76.28, 65.13, 16.43, -2.09.

[0089] Step (2): Synthesis of 4-fluoro-N-(3-(pyridin-3-yl)-1H-indazol-5-yl)benzamide (Compound I-1d)

[0090] Referring to the method described in Step (6) of Reference Example 1, Compound 6d was used to replace Compound 6a in an equimolar amount to obtain Compound I-1d, which is a white solid with a yield of 48%.

[0091] Mp: 266 - 268 °C; 1 1H NMR (300 MHz, DMSO-d6) δ 13.25 (s, 1H), 10.18 (s, 1H), 8.96 (d, J = 1.4 Hz, 1H), 8.46 - 8.33 (m, 2H), 8.11 (dt, J = 8.0, 1.9 Hz, 1H), 7.94 - 7.82 (m, 2H), 7.62 (dd, J = 9.0, 1.8 Hz, 1H), 7.48 - 7.32 (m, 2H), 7.19 (t, J = 8.9 Hz, 2H).; 13 13C NMR (75 MHz, DMSO-d6) δ 166.20, 164.89, 162.90, 149.11, 147.76, 140.82, 139.22, 134.22, 133.80, 131.85, 130.87, 130.75, 130.23, 124.63, 122.11, 120.41, 116.00, 115.71, 111.32, 111.16; HRMS(+ESI) m / z calcd for C 19 H 13 ON4F [M + H] + 332.1073, found 332.1077.

[0092] Example 5

[0093] 3-cyano-N-(3-(pyridin-3-yl)-1H-indazol-5-yl)benzamide (Compound I-1e)

[0094] Step (1): Synthesis of 3-cyano-N-(3-(pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)benzamide (Compound 6e)

[0095] Referring to the method described in Step (5) of Reference Example 1, replacing benzoic acid with an equimolar amount of m-cyanobenzoic acid gave a red-brown oil, which was purified by silica gel column chromatography (eluent: DCM:MeOH = 200:1 V / V) to obtain Compound 6e, a white solid, with a yield of 70%.

[0096] 1 H NMR (300 MHz, DMSO-d6) δ 10.55 (s, 1H), 9.21 - 8.86 (m, 1H), 8.78 - 8.56 (m, 2H), 8.50 - 8.38 (m, 1H), 8.31 (dt, J = 8.0, 1.8 Hz, 2H), 8.09 (dd, J = 7.8, 1.5 Hz, 1H), 7.97 - 7.83 (m, 2H), 7.78 (t, J = 7.9 Hz, 1H), 7.72 - 7.57 (m, 1H), 5.85 (s, 2H), 3.61 (t, J = 7.9 Hz, 2H), 0.83 (t, J = 7.9 Hz, 2H), -0.10 (s, 10H); 13 C NMR (75 MHz, DMSO-d6) δ 164.14, 149.61, 147.87, 141.17, 139.00, 136.33, 135.55, 134.59, 134.27, 133.01, 131.76, 130.41, 129.44, 124.76, 122.36, 121.74, 118.87, 112.06, 111.53, 111.30, 77.49, 66.33, 46.03, 17.62, 9.04, -0.90, -0.97.

[0097] Step (2): Synthesis of 3-cyano-N-(3-(pyridin-3-yl)-1H-indazol-5-yl)benzamide (Compound I-1e)

[0098] Referring to the method described in Step (6) of Reference Example 1, replacing Compound 6a with an equimolar amount of Compound 6e gave Compound I-1e, a white solid, with a yield of 64%.

[0099] Mp: 258 - 260 °C; 11H NMR (300 MHz, DMSO-d6) δ 13.48 (s, 1H), 10.55 (s, 1H), 9.17 (d, J = 2.3 Hz, 1H), 8.71 - 8.54 (m, 2H), 8.46 (s, 1H), 8.31 (dt, J = 6.3, 1.9 Hz, 2H), 8.08 (d, J = 7.8 Hz, 1H), 7.91 - 7.71 (m, 2H), 7.71 - 7.51 (m, 2H); 13 13C NMR (75 MHz, DMSO-d6) δ 164.81, 149.88, 148.48, 141.58, 140.08, 137.14, 136.18, 134.97, 134.21, 133.70, 132.44, 131.09, 130.90, 125.38, 122.70, 121.11, 119.61, 112.74, 112.20, 112.01; HRMS(+ESI) m / z calcd for C 20 H 13 N5O [M + H] + 339.1120, found 339.1120.

[0100] Example 6

[0101] 4-Cyano-N-(3-(pyridin-3-yl)-1H-indazol-5-yl)benzamide (Compound I-1f)

[0102] Step (1): Synthesis of 4-cyano-N-(3-(pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)benzamide (Compound 6f)

[0103] Referring to the method described in Step (5) of Reference Example 1, p-cyanobenzoic acid was used to replace benzoic acid in equimolar amounts to obtain Compound 6f, a reddish-brown oil, which was used directly in the next reaction without purification.

[0104] Step (2): Synthesis of 4-cyano-N-(3-(pyridin-3-yl)-1H-indazol-5-yl)benzamide (I-1f)

[0105] Referring to the method described in Step (6) of Reference Example 1, Compound 6f was used to replace Compound 6a in equimolar amounts to obtain Compound I-1f, a white solid, with a yield of 56%.

[0106] Mp: 254 - 256 °C; 11H NMR (300 MHz, DMSO-d6) δ 13.47 (s, 1H), 10.60 (s, 1H), 9.17 (d, J = 2.3 Hz, 1H), 8.69 - 8.55 (m, 2H), 8.31 (dt, J = 8.0, 2.0 Hz, 1H), 8.21 - 8.13 (m, 2H), 8.09 - 8.01 (m, 2H), 7.82 (dd, J = 9.0, 1.9 Hz, 1H), 7.69 - 7.55 (m, 2H); 13 13C NMR (75 MHz, DMSO-d6) δ 164.34, 148.78, 147.41, 140.64, 139.26, 139.14, 134.21, 133.31, 132.79, 130.06, 128.79, 124.51, 121.82, 120.20, 118.67, 114.11, 111.23, 111.15; HRMS(+ESI) m / z calcd for C 20 H 13 N5O [M + H] + 339.1120, found 339.1116.

[0107] Example 7

[0108] N-(3-(Pyridin-3-yl)-1H-indazol-5-yl)isophthalamide (Compound I-1g)

[0109]

[0110] Procedure: Dissolve Compound I-1e (0.06 g, 0.18 mmol) in 2 mL of dimethyl sulfoxide, and successively add 30% aqueous hydrogen peroxide solution (0.069 mL, 0.884 mmol) and potassium carbonate (0.098 g, 0.71 mmol). Stir at room temperature for 2 h. At this time, the starting material spot disappears by TLC monitoring, and the reaction is complete. Stop the reaction. Add water to the reaction solution, and a large amount of white solid precipitates. Filter by suction, wash the filter cake repeatedly with water, and dry it to obtain Compound I-1g (pale yellow solid, 0.03 g, yield 48%).

[0111] Mp: 288 - 290 °C; 11H NMR (300 MHz, DMSO-d6) δ 13.46 (s, 1H), 10.48 (s, 1H), 9.17 (d, J = 2.3 Hz, 1H), 8.62 (dd, J = 5.1, 1.7 Hz, 2H), 8.50 (s, 1H), 8.32 (dt, J = 8.0, 2.0 Hz, 1H), 8.19 - 8.00 (m, 3H), 7.85 (dd, J = 9.0, 1.8 Hz, 1H), 7.69 - 7.50 (m, 4H); 13 13C NMR (75 MHz, DMSO-d6) δ 167.94, 165.64, 149.15, 147.80, 140.86, 139.26, 135.67, 135.04, 134.21, 133.86, 130.83, 130.25, 128.98, 128.07, 127.38, 124.64, 122.04, 120.45, 111.38, 111.12; HRMS (+ESI) m / z calcd for C 20 H 15 N5O2 [M + H] + 357.1226, found 357.1228.

[0112] Example 8

[0113] N-(3-(Pyridin-3-yl)-1H-indazol-5-yl)terephthalamide (Compound I-1h)

[0114] Referring to the method described in Reference Example 7, Compound I-1e was replaced with an equimolar amount of Compound I-1f to obtain Compound I-1h, a white solid, with a yield of 33%.

[0115] Mp: 286 - 288 °C; 1 1H NMR (300 MHz, DMSO-d6) δ 10.48 (s, 1H), 9.18 (d, J = 2.2 Hz, 1H), 8.63 (d, J = 4.7 Hz, 2H), 8.32 (d, J = 8.0 Hz, 1H), 8.17 (s, 1H), 8.05 (q, J = 8.2 Hz, 4H), 7.85 (d, J = 9.0 Hz, 1H), 7.72 - 7.51 (m, 3H); 1313C NMR(75MHz,DMSO-d6)δ167.75,165.36,149.14,147.79,140.84,139.30,137.75,137.27,134.23,133.77,130.23,128.09,128.06,124.64,122.07,120.43,111.40,111.21;HRMS(+ESI)m / z calcd for C 20 H 15 N5O2[M+H] + 357.1226,found 357.1227.

[0116] Example 9

[0117] N-(3-(Pyridin-3-yl)-1H-indazol-5-yl)benzothioamide (Compound I-1i)

[0118]

[0119] Lawesson's reagent (0.077 g, 0.19 mmol) and toluene (2 mL) were added to a three-necked flask, and compound I-1a (0.03 g, 0.095 mmol) was added. The mixture was purged with nitrogen three times, refluxed and stirred for 2 h. The reaction was monitored by TLC and was found to be complete. The reaction was stopped and cooled to room temperature. The reaction mixture was added with CH2Cl2 (5 mL), washed with saturated NaHCO3 solution (10 mL), washed with saturated NaCl solution (10 mL × 3), dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure. The obtained residue was triturated, and silica gel column chromatography (PE:EA = 2:1 V / V) was carried out to obtain compound I-1i as a yellow solid (0.015 g) with a yield of 48%.

[0120] Mp: >300 °C; 1 1H NMR(300MHz,DMSO-d6)δ13.58(s,1H),11.90(s,1H),9.19(s,1H),8.78 - 8.55(m,2H),8.34(d,J = 7.1Hz,1H),7.86(dd,J = 16.8,8.1Hz,3H),7.69(d,J = 8.9Hz,1H),7.53(ddt,J = 18.8,14.5,6.3Hz,4H); 1313C NMR (75 MHz, DMSO) δ 197.49, 149.00, 147.50, 142.71, 140.92, 139.98, 134.41, 133.99, 131.00, 129.69, 128.27, 127.79, 124.70, 124.41, 119.70, 115.67, 110.90; HRMS(+ESI) m / z calcd for C 19 H 14 N4S [M + H] + 330.0939, found 330.0941.

[0121] Example 10

[0122] N-(3-(3-Nitrophenyl)-1H-indazol-5-yl)benzamide (Compound I-2a)

[0123]

[0124] Step (1): Synthesis of 3-(3-Nitrophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-amine (Compound 7)

[0125] Compound 4 (2.36 g, 6.89 mmol), 3-nitrophenylboronic acid (1.27 g, 7.58 mmol), Pd(OAc)2 (0.15 g, 0.69 mmol), X-Phos (0.66 g, 1.38 mmol), and K2CO3 (2.86 g, 20.68 mmol) were placed in a three-necked flask. The flask was purged with N2 three times, and 30 mL of a mixed solvent (dioxane:H2O = 7:1 V / V) was added. After stirring at room temperature for 10 min to activate the catalyst, the mixture was heated and stirred at 100 °C. After monitoring the reaction by TLC until completion, heating was stopped, and the reaction solution was cooled to room temperature. It was filtered through diatomaceous earth, and the filtrate was extracted with CH2Cl2 (30 mL × 3). The organic phase was washed with saturated NaHCO3 solution (30 mL × 3) and saturated NaCl solution (30 mL × 3), dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (PE:EA = 8:1 V / V) to obtain Compound 7 (yellow solid, 1.85 g, 4.82 mmol, yield 70%).

[0126] 11H NMR (300 MHz, DMSO-d6) δ 8.67 (s, 1H), 8.34 (d, J = 7.8 Hz, 1H), 8.22 (dd, J = 8.2, 2.3 Hz, 1H), 7.81 (t, J = 8.0 Hz, 1H), 7.54 (d, J = 8.8 Hz, 1H), 7.18 (s, 1H), 6.96 (dd, J = 8.9, 1.9 Hz, 1H), 5.73 (s, 2H), 5.15 (s, 2H), 3.57 (t, J = 7.9 Hz, 2H), 0.82 (t, J = 7.9 Hz, 2H), -0.11 (s, 9H); 13 13C NMR (75 MHz, DMSO-d6) δ 148.70, 144.83, 139.41, 136.17, 135.84, 132.73, 130.98, 122.80, 122.38, 120.71, 119.06, 111.57, 100.52, 77.33, 66.03, 17.50, -1.00. HRMS(+ESI) m / z calcd for C 19 H 24 O3Si [M + H] + 384.1618, found 445.2764.

[0127] Step (2): Synthesis of N-(3-(3-nitrophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)benzamide (Compound 8a)

[0128] Benzoic acid (0.063 g, 0.52 mmol) and a catalytic amount of 4-dimethylaminopyridine (0.006 g, 0.051 mmol) were added to dichloromethane (3 mL). Carbodiimide hydrochloride (0.20 g, 0.52 mmol) was added in portions, and the mixture was stirred at room temperature for 10 min. Compound 7 (0.17 g, 0.51 mmol) was added, and the reaction was carried out at room temperature. The reaction was monitored by TLC until it was complete, and then the reaction was stopped. The reaction mixture was washed successively with saturated NaHCO3 solution (30 mL × 3), saturated NaCl solution (30 mL × 3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain a red-brown oily compound. Trituration and silica gel column chromatography (PE:EA = 6:1 V / V) were carried out to obtain Compound 8a, a pale yellow oily substance, with a yield of 89%.

[0129] 11H NMR (300 MHz, DMSO-d6) δ 10.47 (s, 1H), 8.73 (t, J = 2.0 Hz, 1H), 8.64 (s, 1H), 8.40 (dt, J = 7.8, 1.3 Hz, 1H), 8.30 (ddd, J = 8.2, 2.4, 1.0 Hz, 1H), 8.05 - 7.97 (m, 2H), 7.95 - 7.84 (m, 3H), 7.64 - 7.53 (m, 3H), 5.86 (s, 2H), 3.62 (t, J = 7.9 Hz, 2H), 0.84 (t, J = 7.9 Hz, 2H), -0.10 (s, 9H); 13 13C NMR (75 MHz, DMSO-d6) δ 165.94, 148.61, 141.29, 138.80, 135.15, 134.88, 134.70, 132.94, 131.92, 131.05, 128.73, 127.94, 122.90, 122.44, 121.30, 121.06, 111.09, 110.92, 77.31, 66.13, 17.39, -1.14.

[0130] Step (3): Synthesis of N-(3-(3-nitrophenyl)-1H-indazol-5-yl)benzamide (Compound I-2a)

[0131] Dissolve Compound 8a (0.126 g, 0.258 mmol) in tetrahydrofuran (3 mL). While stirring, add ethylenediamine (0.225 mL, 0.774 mmol) and tetrabutylammonium fluoride solution (1 M, 1.29 mL, 1.29 mmol, prepared from THF) in sequence. Reflux the reaction until TLC monitoring shows that the reaction is complete. Stop heating and cool to room temperature. Distill off part of the solvent under reduced pressure, add 10 mL of ethyl acetate, wash with saturated NaHCO3 solution (15 mL × 3) and saturated NaCl solution (15 mL × 3) in sequence, dry over anhydrous Na2SO4, filter, and concentrate under reduced pressure to obtain a pale yellow crude product. After dissolving the crude product in a small amount of tetrahydrofuran, add a large amount of petroleum ether. A white solid precipitates. Filter by suction, wash the filter cake with a small amount of ethyl acetate, and dry to obtain Compound I-2a (0.069 mg, yield 75%).

[0132] Mp: 242 - 244 °C; 1 1H NMR (300 MHz, DMSO-d6) δ 13.55 (s, 1H), 10.42 (s, 1H), 8.74 (t, J = 2.0

[0133] Hz, 1H), 8.63 (s, 1H), 8.41 (d, J = 7.8 Hz, 1H), 8.26 (dd, J = 8.0, 2.0 Hz, 1H), 8.00 (d, J = 6.6 Hz, 2H), 7.91 - 7.80 (m, 2H), 7.69 - 7.51 (m, 4H); 13 C NMR (75 MHz, DMSO-d6) δ 165.83, 148.57, 141.03, 139.12, 135.67, 135.22, 133.92, 132.67, 131.77, 130.87, 128.65, 127.87, 122.35, 122.04, 120.70, 119.95, 111.22, 110.62; HRMS(+ESI) m / z calcd for C 20 H 14 O3N4 [M + H] + 358.1066, found 358.1068.

[0134] Example 11

[0135] 2-Fluoro-N-(3-(3-nitrophenyl)-1H-indazol-5-yl)benzamide (Compound I-2b)

[0136] Step (1): Referring to the method described in Step (2) of Example 10, replace benzoic acid with o-fluorobenzoic acid in equimolar amounts to obtain Compound 8b, a pale yellow oil, with a yield of 82%.

[0137] 1 H NMR (300 MHz, DMSO-d6) δ 10.62 (s, 1H), 8.71 (t, J = 2.0 Hz, 1H), 8.64 (s, 1H), 8.38 (dt, J = 7.9, 1.3 Hz, 1H), 8.29 (ddd, J = 8.3, 2.4, 1.0 Hz, 1H), 7.93 - 7.79 (m, 3H), 7.73 (td, J = 7.4, 1.7 Hz, 1H), 7.66 - 7.54 (m, 1H), 7.45 - 7.31 (m, 2H), 5.85 (s, 2H), 3.61 (t, J = 7.9 Hz, 2H), 0.84 (t, 2H), -0.10 (s, 9H); 1313C NMR (75 MHz, DMSO-d6) δ 162.19, 147.65, 140.36, 137.84, 133.84, 133.50, 131.99, 130.12, 129.30, 123.95, 122.00, 120.73, 120.31, 120.09, 115.69, 115.41, 110.39, 109.22, 76.36, 65.17, 16.41, -2.09.

[0138] Step (2): Synthesis of 2-fluoro-N-(3-(3-nitrophenyl)-1H-indazol-5-yl)benzamide (Compound I-2b)

[0139] Referring to the method described in Step (3) of Reference Example 10, Compound 8b was used to replace Compound 8a in equimolar amounts to obtain Compound I-2b, a white solid, with a yield of 72%.

[0140] Mp: 206 - 208 °C; 1 1H NMR (300 MHz, DMSO-d6) δ 13.57 (s, 1H), 10.57 (s, 1H), 8.73 (t, J = 2.0

[0141] Hz, 1H), 8.63 (s, 1H), 8.40 (d, J = 7.6 Hz, 1H), 8.26 (ddd, J = 8.4, 2.5, 1.1 Hz, 1H), 7.86 (t, J = 8.0 Hz, 1H), 7.79 - 7.54 (m, 4H), 7.43 - 7.30 (m, 2H); 13 13C NMR (75 MHz, DMSO-d6) δ 163.09, 160.85, 157.55, 148.63, 141.11, 139.17, 135.68, 133.78, 132.75, 130.97, 130.26, 125.43, 125.23, 124.92, 124.87, 122.47, 121.33, 120.75, 119.97, 116.64, 116.35, 111.55, 109.90; HRMS(+ESI) m / z calcd for C 20 H 13 O3N4F [M + H] + 376.0972, found 376.0978.

[0142] Example 12

[0143] 3-fluoro-N-(3-(3-nitrophenyl)-1H-indazol-5-yl)benzamide (Compound I-2c)

[0144] Step (1): Synthesis of 3-fluoro-N-(3-(3-nitrophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)benzamide (Compound 8c)

[0145] Referring to the method described in Step (2) of Reference Example 10, replace benzoic acid with m-fluorobenzoic acid in equimolar amounts to obtain Compound 8c, a colorless oil, with a yield of 84%.

[0146] 1 H NMR (300 MHz, DMSO-d6) δ 10.51 (s, 1H), 8.71 (t, J = 2.0 Hz, 1H), 8.62 (s, 1H), 8.38 (d, J = 7.8 Hz, 1H), 8.28 (dd, J = 8.3, 1.4 Hz, 1H), 7.94 - 7.78 (m, 5H), 7.61 (td, J = 8.0, 5.8 Hz, 1H), 7.51 - 7.41 (m, 1H), 5.85 (s, 2H), 3.61 (t, J = 7.9 Hz, 2H), 0.84 (t, J = 7.9 Hz, 2H), -0.11 (s, 9H); 13 C NMR (75 MHz, DMSO-d6) δ 164.71, 164.08, 148.83, 141.54, 139.09, 137.68, 137.59, 135.03, 134.59, 133.16, 131.28, 131.20, 131.09, 124.39, 124.36, 123.16, 122.64, 121.47, 121.28, 119.18, 118.90, 115.13, 114.83, 111.36, 77.52, 66.35, 17.59.

[0147] Step (2): Synthesis of 3-fluoro-N-(3-(3-nitrophenyl)-1H-indazol-5-yl)benzamide (Compound I-2c)

[0148] Referring to the method described in Step (3) of Reference Example 10, replace Compound 8a with Compound 8c in equimolar amounts to obtain Compound I-2c, a white solid, with a yield of 66%.

[0149] Mp: 196 - 198 °C; 11H NMR (300 MHz, DMSO-d6) δ 13.60 (s, 1H), 10.51 (s, 1H), 8.74 (s, 1H), 8.63 (s, 1H), 8.41 (d, J = 7.8 Hz, 1H), 8.26 (dd, J = 8.3, 2.4 Hz, 1H), 7.85 (dd, J = 13.1, 8.5 Hz, 4H), 7.64 (dd, J = 15.9, 7.8 Hz, 2H), 7.46 (td, J = 8.4, 2.6 Hz, 1H); 13 13C NMR (75 MHz, DMSO-d6) δ 164.67, 164.08, 160.84, 148.85, 141.31, 139.47, 137.79, 137.70, 135.92, 133.90, 132.96, 131.17, 131.05, 124.40, 122.65, 122.31, 120.97, 120.17, 119.09, 118.82, 115.16, 114.86, 111.56, 111.10; HRMS(+ESI) m / z calcd for C 20 H 13 O3N4F [M + H] + 376.0972, found 376.0978.

[0150] Example 13

[0151] 4-Fluoro-N-(3-(3-nitrophenyl)-1H-indazol-5-yl)benzamide (Compound I-2d)

[0152] Step (1): Synthesis of 4-fluoro-N-(3-(3-nitrophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)benzamide (Compound 8d)

[0153] Referring to the method described in Step (2) of Reference Example 10, p-fluorobenzoic acid was used to replace benzoic acid in an equimolar amount to obtain Compound 8d, a pale yellow oil, with a yield of 86%.

[0154] 11H NMR (300 MHz, DMSO-d6) δ 10.46 (s, 1H), 8.71 (t, J = 2.0 Hz, 1H), 8.61 (s, 1H), 8.39 (d, J = 8.0 Hz, 1H), 8.28 (ddd, J = 8.3, 2.4, 1.0 Hz, 1H), 8.14 - 8.03 (m, 2H), 7.93 - 7.79 (m, 3H), 7.39 (t, J = 8.9 Hz, 2H), 5.85 (s, 2H), 3.61 (t, J = 8.0 Hz, 2H), 0.84 (t, J = 8.0 Hz, 2H), -0.10 (s, 9H); 13 13C NMR (75 MHz, DMSO-d6) δ 164.80, 162.76, 148.61, 141.28, 138.81, 134.89, 134.60, 132.93, 131.58, 131.02, 130.74, 130.62, 122.89, 122.45, 121.30, 121.07, 115.81, 115.52, 111.09, 111.00, 77.32, 66.14, 17.39, -1.14.

[0155] Step (2): 4-Fluoro-N-(3-(3-nitrophenyl)-1H-indazol-5-yl)benzamide (Compound I-2d)

[0156] Referring to the method described in Step (3) of Reference Example 10, replace Compound 8a with an equimolar amount of Compound 8d to obtain I-2d, a white solid, with a yield of 68%.

[0157] Mp: 236 - 238 °C; 1 1H NMR (300 MHz, DMSO-d6) δ 13.56 (s, 1H), 10.44 (s, 1H), 8.73 (s, 1H), 8.61 (s, 1H), 8.41 (d, J = 7.8 Hz, 1H), 8.26 (dd, J = 8.2, 2.3 Hz, 1H), 8.09 (dd, J = 8.6, 5.7 Hz, 2H), 7.92 - 7.77 (m, 2H), 7.66 (d, J = 9.0 Hz, 1H), 7.39 (t, J = 8.8 Hz, 2H).; 1313C NMR(75MHz,DMSO-d6)δ164.58,162.51,148.45,140.91,139.01,135.52,133.66,132.56,131.50,130.76,130.52,130.40,122.25,121.95,120.57,119.80,115.61,115.32,111.11,110.59;HRMS(+ESI)m / z calcd for C 20 H 13 O3N4F[M+H] + 376.0972,found 376.0973.

[0158] Example 14

[0159] 3-Cyano-N-(3-(3-nitrophenyl)-1H-indazol-5-yl)benzamide (Compound I-2e)

[0160] Step (1): Synthesis of 3-cyano-N-(3-(3-nitrophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)benzamide (Compound 8e)

[0161] Referring to the method described in Step (2) of Reference Example 10, benzoic acid was replaced with isomolar m-cyanobenzoic acid to obtain Compound 8e, a pale yellow oil, with a yield of 85%.

[0162] 1 1H NMR(300MHz,DMSO-d6)δ10.63(s,1H),8.71(t,J=2.0Hz,1H),8.62(s,1H),8.45(s,1H),8.38(d,J=7.8Hz,1H),8.29(td,J=6.3,3.0Hz,2H),8.12-8.04(m,1H),7.93-7.83(m,3H),7.77(t,J=7.8Hz,1H),5.85(s,2H),3.62(t,J=7.9Hz,2H),0.84(t,J=7.9Hz,2H),-0.10(s,10H); 1313C NMR (75 MHz, DMSO-d6) δ 164.33, 148.95, 141.68, 139.25, 136.44, 135.66, 135.11, 134.59, 133.29, 133.14, 131.88, 131.42, 130.52, 123.32, 122.64, 121.57, 121.41, 118.98, 112.16, 111.58, 111.41, 77.65, 66.47, 17.70, -0.80.

[0163] Step (2): Synthesis of 3-cyano-N-(3-(3-nitrophenyl)-1H-indazol-5-yl)benzamide (Compound I-2e)

[0164] Referring to the method described in Step (3) of Reference Example 10, Compound 8e was used to replace Compound 8a in equimolar amounts to obtain Compound I-2e, a white solid, with a yield of 70%.

[0165] Mp: 232 - 234 °C; 1 1H NMR (300 MHz, DMSO-d6) δ 13.58 (s, 1H), 10.65 (s, 1H), 8.73 (s, 1H), 8.63 (s, 1H), 8.41 (d, J = 7.8 Hz, 1H), 8.27 (d, J = 8.3 Hz, 1H), 8.16 (d, J = 7.9 Hz, 2H), 8.06 (d, J = 8.1 Hz, 2H), 7.85 (dd, J = 16.3, 8.3 Hz, 2H), 7.67 (d, J = 8.9 Hz, 1H); 13 13C NMR (75 MHz, DMSO-d6) δ 164.27, 148.94, 141.49, 139.61, 136.54, 135.97, 135.58, 133.86, 133.12, 133.07, 131.86, 131.26, 130.47, 122.79, 122.27, 121.09, 120.28, 118.99, 112.14, 111.74, 111.17; HRMS(+ESI) m / z calcd for C 21 H 13 O3N5[M + H] + 383.1018, found 383.1022.

[0166] Example 15

[0167] 4-cyano-N-(3-(3-nitrophenyl)-1H-indazol-5-yl)benzamide (Compound I-2f)

[0168] Step (1): Synthesis of 4-cyano-N-(3-(3-nitrophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)benzamide (Compound 8f)

[0169] Referring to the method described in Step (2) of Reference Example 10, equimolar replacement of benzoic acid with p-cyanobenzoic acid gave Compound 8f, a pale yellow oil, with a yield of 81%.

[0170] 1 H NMR (300 MHz, DMSO-d6) δ 10.72 (s, 1H), 8.70 (s, 1H), 8.63 (s, 1H), 8.38 (d, J = 7.9 Hz, 1H), 8.32 - 8.25 (m, 1H), 8.17 (d, J = 8.2 Hz, 2H), 8.04 (d, J = 8.1 Hz, 2H), 7.96 - 7.82 (m, 3H), 5.85 (s, 2H), 3.61 (t, J = 7.9 Hz, 2H), 0.84 (t, J = 7.9 Hz, 2H), -0.11 (s, 9H).; 13 C NMR (75 MHz, DMSO-d6) δ 164.52, 148.64, 141.40, 139.17, 138.97, 134.84, 134.30, 132.99, 132.85, 131.09, 128.87, 122.99, 122.40, 121.30, 121.12, 118.69, 114.24, 111.23, 77.38, 66.21, 17.44, -1.08.

[0171] Step (2): 4-cyano-N-(3-(3-nitrophenyl)-1H-indazol-5-yl)benzamide (Compound I-2f)

[0172] Referring to the method described in Step 3 of Reference Example 10, equimolar replacement of Compound 8a with Compound 8f gave Compound I-2f, a white solid, with a yield of 64%.

[0173] Mp: 278 - 280 °C; 1 H NMR (300 MHz, DMSO-d6) δ 10.55 (s, 1H), 8.76 (s, 1H), 8.66 (s, 1H), 8.44 (d, J = 7.8 Hz, 1H), 8.28 (dd, J = 8.1, 2.3 Hz, 1H), 8.19 (s, 1H), 8.07 (q, J = 8.2 Hz, 4H), 7.94 - 7.82 (m, 2H), 7.69 (d, J = 9.0 Hz, 1H), 7.60 (s, 1H); 1313C NMR(75MHz, DMSO-d6) δ 164.61, 148.80, 141.34, 139.44, 135.80, 133.69, 132.96, 131.13, 128.98, 122.66, 122.19, 120.94, 120.11, 118.82, 114.27, 111.58, 111.12; HRMS(+ESI) m / z calcd for C 21 H 13 O3N5[M + H] + 383.1018, found 383.1016.

[0174] Example 16

[0175] N-(3-(3-Nitrophenyl)-1H-indazol-5-yl) isophthalamide (Compound I-2g)

[0176] Referring to the method described in Reference Example 7, Compound I-1e was replaced with an equimolar amount of Compound I-2e to obtain Compound I-2g, a white solid, with a yield of 58%.

[0177] Mp: >300 °C; 1 1H NMR(300MHz, DMSO-d6) δ 13.60(s, 1H), 10.57(s, 1H), 8.76(s, 1H), 8.66

[0178] (s, 1H), 8.59 - 8.38(m, 2H), 8.28(dd, J = 8.2, 2.3Hz, 1H), 8.22 - 8.04(m, 3H), 7.88(t, J = 8.1Hz, 2H), 7.76 - 7.50(m, 3H); 13 13C NMR(75MHz, DMSO-d6) δ 167.84, 165.66, 148.77, 141.22, 139.36, 135.82, 135.59, 134.94, 133.97, 132.89, 131.11, 130.79, 128.92, 127.32, 122.61, 122.17, 120.88, 120.10, 111.50, 110.82; HRMS(+ESI) m / z calcd for C 21 H 15 O4N5[M + H] + 401.1124, found 401.1124.

[0179] Example 17

[0180] N-(3-(3-Nitrophenyl)-1H-indazol-5-yl)terephthalamide (Compound I-2h)

[0181] Referring to the method described in Reference Example 7, Compound I-1e was replaced with an equimolar amount of Compound I-2f to obtain Compound I-2h, a white solid, with a yield of 65%.

[0182] Mp: 282 - 284 °C; 1 H NMR (300 MHz, DMSO-d6) δ 10.55 (s, 1H), 8.76 (s, 1H), 8.66 (s, 1H), 8.44 (d, J = 7.8 Hz, 1H), 8.28 (dd, J = 8.1, 2.3 Hz, 1H), 8.19 (s, 1H), 8.07 (q, J = 8.2 Hz, 4H), 7.94 - 7.82 (m, 2H), 7.69 (d, J = 9.0 Hz, 1H), 7.60 (s, 1H); 13 C NMR (75 MHz, DMSO-d6) δ 167.60, 165.35, 148.75, 141.13, 139.59, 137.67, 137.16, 135.90, 133.85, 132.82, 131.07, 128.02, 127.96, 122.49, 122.06, 120.84, 120.14, 111.67, 110.89; HRMS (+ESI) m / z calcd for C 21 H 15 O4N5[M + H] + 401.1124, found 401.1121.

[0183] Example 18

[0184] N-(3-(3-Nitrophenyl)-1H-indazol-5-yl)benzothioamide (Compound I-2i)

[0185] Referring to the method described in Reference Example 9, Compound I-2a was replaced with an equimolar amount of Compound I-1a. After concentration under reduced pressure, a yellow solid was obtained. It was triturated and purified by silica gel column chromatography (PE:EA = 3:1 V / V) to obtain Compound I-2i, a yellow solid, with a yield of 61%.

[0186] Mp: 254 - 256 °C; 11H NMR (300 MHz, DMSO-d6) δ 13.67 (s, 1H), 11.92 (s, 1H), 8.82 - 8.64 (m, 2H), 8.44 (d, J = 7.8 Hz, 1H), 8.25 (dd, J = 8.2, 2.4 Hz, 1H), 7.96 - 7.77 (m, 4H), 7.71 (d, J = 8.9 Hz, 1H), 7.52 (dq, J = 14.1, 6.9 Hz, 3H); 13 13C NMR (75 MHz, DMSO-d6) δ 198.14, 161.70, 148.97, 143.08, 141.84, 140.56, 135.66, 135.09, 133.15, 133.04, 132.89, 131.42, 131.33, 128.69, 128.13, 125.31, 122.93, 121.14, 119.92, 115.78, 114.18, 113.98, 111.51; HRMS (+ESI) m / z calcd for C 20 H 14 O2N4S [M + H] + 374.0837, found 374.0835.

[0187] Example 19

[0188] 4 - ((3 - (Pyridin - 3 - yl) - 1H - indazol - 5 - yl)amino) - N - (tetrahydropyran - 4 - yl)benzamide (Compound II - 1a)

[0189]

[0190] Step (1): Synthesis of 4 - bromo - N - (tetrahydropyran - 4 - yl)benzamide (Compound 10a)

[0191] Compound 9 (1 g, 4.97 mmol) was added to 3 mL of dichloromethane. Catalytic amounts of 4 - dimethylaminopyridine (0.061 g, 0.50 mmol) and carbodiimide hydrochloride (1.05 g, 5.47 mmol) were added successively. After stirring at room temperature for 10 min, 4 - aminotetrahydropyran (0.559 mL, 4.97 mmol) was added. Stirring was continued at room temperature until TLC showed complete reaction, and then the reaction was stopped. The reaction mixture was washed with 1 M HCl solution (10 mL × 3), extracted with dichloromethane (10 mL × 3), the organic phase was washed with saturated NaCl solution (10 mL × 3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to remove the solvent, obtaining Compound 10a (white solid, 1.39 g, 4.89 mmol, yield 99%).

[0192] 1 1H NMR (300 MHz, DMSO-d6) δ 8.41 (d, J = 7.7 Hz, 1H), 7.85 - 7.60 (m, 4H), 4.07 - 3.78 (m, 3H), 1.74 (ddd, J = 12.3, 4.5, 2.0 Hz, 2H), 1.56 (qd, J = 11.9, 4.4 Hz, 2H). 1 1H NMR (300 MHz, DMSO-d6 + D2O) δ 7.87 - 7.63 (m, 4H), 3.43 (td, J = 11.8, 2.2 Hz, 2H), 1.87 - 1.74 (m, 2H), 1.62 (qd, J = 11.9, 4.4 Hz, 2H).

[0193] Step (2): 4 - ((3-(Pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)amino)-N-(tetrahydropyran-4-yl)benzamide (Compound 11a)

[0194] Compound 5 (0.13 g, 0.38 mmol), Compound 10a (0.108 g, 0.38 mmol), tris(dibenzylideneacetone)dipalladium (0.035 g, 0.039 mmol), X-Phos (0.036 g, 0.076 mmol), and potassium carbonate (0.21 g, 1.53 mmol) were placed in a three-necked flask. The flask was purged with nitrogen three times, and then refluxed with stirring at 100 °C for 4 h. TLC monitoring showed that the reaction was complete at this time. Heating was stopped, and the reaction solution was cooled to room temperature. It was filtered through diatomaceous earth, and the filtrate was extracted with CH2Cl2 (15 mL × 3). The organic phase was washed successively with saturated NaHCO3 solution (20 mL × 3) and saturated NaCl solution (30 mL × 3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain Compound 11a (brown oil), which was used directly in the next step without purification.

[0195] Step (3): Synthesis of 4 - ((3-(Pyridin-3-yl)-1H-indazol-5-yl)amino)-N-(tetrahydropyran-4-yl)benzamide (Compound II-1a)

[0196] Referring to the method described in Step (6) of Reference Example 1, Compound 6a was replaced with an equimolar amount of Compound 11a to obtain Compound II-1a, a white solid, and the yield of the two-step reaction was 26%.

[0197] Mp: 264 - 266 °C; 11H NMR (300 MHz, DMSO-d6) δ 13.38 (s, 1H), 9.12 (s, 1H), 8.64 - 8.47 (m, 2H), 8.29 (d, J = 7.9 Hz, 1H), 7.99 (d, J = 7.8 Hz, 1H), 7.74 (d, J = 9.8 Hz, 3H), 7.61 (d, J = 8.8 Hz, 1H), 7.54 (dd, J = 8.1, 4.9 Hz, 1H), 7.29 (d, J = 8.9 Hz, 1H), 7.01 (d, J = 8.3 Hz, 2H), 4.07 - 3.79 (m, 3H), 1.83 - 1.42 (m, 4H); 1 1H NMR (300 MHz, DMSO-d6 + CD3OD) δ 9.12 (d, J = 2.2 Hz, 1H), 8.59 (dd, J = 4.8, 1.6 Hz, 1H), 8.31 (dt, J = 8.0, 1.9 Hz, 1H), 8.08 (d, J = 7.8 Hz, 1H), 7.74 (dd, J = 9.1, 2.0 Hz, 3H), 7.64 (d, J = 8.9 Hz, 1H), 7.60 - 7.51 (m, 1H), 7.32 (dd, J = 8.9, 1.9 Hz, 1H), 7.07 - 6.98 (m, 2H), 3.93 - 3.82 (m, 2H), 3.38 (dd, J = 12.7, 10.4 Hz, 2H), 1.74 (dd, J = 11.7, 4.0 Hz, 2H), 1.57 (qd, J = 11.9, 4.2 Hz, 2H); 13 13C NMR (75 MHz, DMSO-d6) δ 166.07, 148.92, 148.21, 147.60, 140.22, 138.58, 136.51, 134.44, 130.23, 129.48, 124.74, 124.37, 122.85, 121.07, 113.76, 112.26, 109.22, 66.76, 45.97, 32.97; HRMS (+ESI) m / z calcd for C 24 H 23 N5O2 [M + H] + 413.1852, found 413.1851.

[0198] Example 20

[0199] N-Cyclohexyl-4-((3-(pyridin-3-yl)-1H-indazol-5-yl)amino)benzamide (Compound II-1b)

[0200] Step (1): Synthesis of 4-bromo-N-cyclohexylbenzamide (Compound 10b)

[0201] Referring to the method described in step (1) of Reference Example 19, cyclohexylamine was used to replace 4-aminotetrahydropyran in an equimolar amount to obtain Compound 10b, a white solid, with a yield of 98%.

[0202] 1 H NMR (300 MHz, DMSO-d6) δ 8.30 (s, 1H), 7.95 - 7.50 (m, 4H), 3.74 (s, 1H), 1.91 - 1.48 (m, 5H), 1.29 (s, 5H).

[0203] Step (2): N-Cyclohexyl-4-((3-(pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)amino)benzamide (Compound 11b)

[0204] Referring to the method described in step (2) of Reference Example 19, Compound 10b was used to replace Compound 10a in an equimolar amount to obtain Compound 11b (a brown oil), which was used directly in the next reaction without purification.

[0205] Step (3): Synthesis of N-Cyclohexyl-4-((3-(pyridin-3-yl)-1H-indazol-5-yl)amino)benzamide (Compound II-1b)

[0206] Referring to the method described in step (6) of Reference Example 1, Compound 11b was used to replace Compound 6a in an equimolar amount to obtain Compound II-1b, a white solid, with a two-step reaction yield of 19%.

[0207] Mp: 264 - 266 °C; 1 H NMR (300 MHz, DMSO-d6) δ 13.39 (s, 1H), 9.12 (d, J = 2.3 Hz, 1H),

[0208] 8.62 - 8.49 (m, 2H), 8.29 (dt, J = 8.0, 2.0 Hz, 1H), 7.87 (d, J = 8.0 Hz, 1H), 7.78 - 7.67 (m, 3H), 7.65 - 7.50 (m, 2H), 7.34 - 7.25 (m, 1H), 7.00 (d, J = 8.6 Hz, 2H), 3.73 (s, 1H), 1.68 (dd, J = 45.2, 16.1 Hz, 5H), 1.37 - 1.07 (m, 5H); 1313C NMR(75MHz,DMSO-d6)δ165.69,149.05,148.28,147.87,140.38,138.90,136.92,134.52,129.66,125.03,124.83,122.91,121.35,113.98,112.47,109.21,48.78,33.33,26.03,25.77;HRMS(+ESI)m / z calcd for C 25 H 25 N5O[M+H] + 411.2059,found 411.2059.

[0209] Example 21

[0210] N-Phenyl-4-((3-(pyridin-3-yl)-1H-indazol-5-yl)amino)benzamide (Compound II-1c)

[0211] Step (1): Synthesis of 4-bromo-N-phenylbenzamide (Compound 10c)

[0212] Referring to the method described in Step (1) of Reference Example 19, aniline was used to replace 4-aminotetrahydropyran in an equimolar amount to obtain Compound 10c, a white solid, with a yield of 96%.

[0213] 1 1H NMR(300MHz,DMSO-d6)δ10.48(s,1H),8.91(d,J = 2.5Hz,1H),8.23(dd,J = 8.3,2.6Hz,1H),7.85(d,J = 8.3Hz,1H),7.75(d,J = 8.0Hz,2H),7.37(t,J = 7.8Hz,2H),7.13(t,J = 7.4Hz,1H).

[0214] Step (2): N-Phenyl-4-((3-(pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)amino)benzamide (Compound 11c)

[0215] Referring to the method described in Step (2) of Reference Example 19, Compound 10c was used to replace Compound 10a in an equimolar amount to obtain Compound 11c (a brown oil), which was used directly in the next reaction without purification.

[0216] Step (3): Synthesis of N-Phenyl-4-((3-(pyridin-3-yl)-1H-indazol-5-yl)amino)benzamide (Compound II-1c)

[0217] Referring to the method described in step (6) of Reference Example 1, compound 6a was replaced with an equimolar amount of compound 11c to obtain compound II-1c, a white solid, with a two-step reaction yield of 33%.

[0218] Mp: 234 - 236 °C; 1 1H NMR (300 MHz, DMSO-d6) δ 13.47 (s, 1H), 10.00 (s, 1H), 9.20 (s, 1H), 8.74 (s, 1H), 8.65 (d, J = 4.2 Hz, 1H), 8.37 (dt, J = 7.9, 1.9 Hz, 1H), 7.93 (d, J = 8.7 Hz, 2H), 7.90 - 7.78 (m, 3H), 7.70 (d, J = 8.9 Hz, 1H), 7.61 (dd, J = 8.1, 4.8 Hz, 1H), 7.38 (t, J = 7.9 Hz, 3H), 7.18 - 7.06 (m, 3H); 13 13C NMR (75 MHz, DMSO-d6) δ 165.56, 149.00, 148.88, 147.81, 140.36, 140.13, 138.88, 136.35, 134.32, 130.32, 130.05, 129.05, 124.65, 124.40, 123.64, 122.97, 121.18, 120.69, 113.73, 112.32, 109.79. HRMS (+ESI) m / z calcd for C 25 H 19 N5O [M + H] + 405.1590, found 405.1590.

[0219] Example 22

[0220] 4 - ((3-(3-Nitrophenyl)-1H-indazol-5-yl)amino)-N-(tetrahydropyran-4-yl)benzamide (Compound II-2a)

[0221] Step (1): Synthesis of 4 - ((3-(3-nitrophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)amino)-N-(tetrahydropyran-4-yl)benzamide (Compound 12a)

[0222] Referring to the method described in step (2) of Reference Example 19, compound 5 was replaced with an equimolar amount of compound 7 to obtain compound 12a (a brown oil), which was used directly in the next step without purification.

[0223] Step (2): Synthesis of 4-((3-(3-nitrophenyl)-1H-indazol-5-yl)amino)-N-(tetrahydropyran-4-yl)benzamide (Compound II-2a)

[0224] Referring to the method described in Step (6) of Reference Example 1, Compound 12a was used to replace Compound 6a in equimolar amounts to obtain Compound II-2a, a white solid, and the yield of the two-step reaction was 24%.

[0225] Mp: 222 - 224 °C; 1 H NMR (300 MHz, DMSO-d6) δ 13.38 (s, 1H), 9.12 (s, 1H), 8.64 - 8.47 (m, 2H), 8.29 (d, J = 7.9 Hz, 1H), 7.99 (d, J = 7.8 Hz, 1H), 7.74 (d, J = 9.8 Hz, 3H), 7.61 (d, J = 8.8 Hz, 1H), 7.54 (dd, J = 8.1, 4.9 Hz, 1H), 7.29 (d, J = 8.9 Hz, 1H), 7.01 (d, J = 8.3 Hz, 2H), 4.07 - 3.79 (m, 3H), 1.83 - 1.42 (m, 4H); 1 H NMR (300 MHz, DMSO-d6 + D2O) δ 9.12 (d, J = 2.2 Hz, 1H), 8.59 (dd, J = 4.8, 1.6 Hz, 1H), 8.31 (dt, J = 8.0, 1.9 Hz, 1H), 8.08 (d, J = 7.8 Hz, 1H), 7.74 (dd, J = 9.1, 2.0 Hz, 3H), 7.64 (d, J = 8.9 Hz, 1H), 7.60 - 7.51 (m, 1H), 7.32 (dd, J = 8.9, 1.9 Hz, 1H), 7.07 - 6.98 (m, 2H), 3.93 - 3.82 (m, 2H), 3.38 (dd, J = 12.7, 10.4 Hz, 2H), 1.74 (dd, J = 11.7, 4.0 Hz, 2H), 1.57 (qd, J = 11.9, 4.2 Hz, 2H); 13 C NMR (75 MHz, DMSO) δ 166.07, 148.92, 148.21, 147.60, 140.22, 138.58, 136.51, 134.44, 130.23, 129.48, 124.74, 124.37, 122.85, 121.07, 113.76, 112.26, 109.22, 66.76, 45.97, 32.97; HRMS(+ESI) m / z calcd for C 25 H 23 O4N5[M + H] +457.1750, found 457.1748.

[0226] Example 23

[0227] N-Cyclohexyl-4-((3-nitrophenyl)-1H-indazol-5-yl)amino)benzamide (Compound II-2b)

[0228] Step (1): N-Cyclohexyl-4-((3-(3-nitrophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)amino)benzamide (Compound 12b)

[0229] Referring to the method described in Step (2) of Reference Example 19, Compound 5 was replaced with an equimolar amount of Compound 7, and Compound 10a was replaced with an equimolar amount of Compound 10b to obtain Compound 12b (brown oil), which was used directly in the next reaction without purification.

[0230] Step (2): Synthesis of N-Cyclohexyl-4-((3-nitrophenyl)-1H-indazol-5-yl)amino)benzamide (Compound II-2b)

[0231] Referring to the method described in Step (6) of Reference Example 1, Compound 6a was replaced with an equimolar amount of Compound 12b to obtain Compound II-2b, a white solid, and the yield of the two-step reaction was 17%.

[0232] Mp: 204-206 °C; 1 1H NMR (300 MHz, DMSO-d6) δ 13.48 (s, 1H), 8.70 (t, J = 2.0 Hz, 1H),

[0233] 8.58 (s, 1H), 8.40 (d, J = 7.8 Hz, 1H), 8.22 (dd, J = 8.0, 1.9 Hz, 1H), 7.91-7.76 (m, 3H), 7.73 (d, J = 8.5 Hz, 2H), 7.63 (d, J = 8.9 Hz, 1H), 7.32 (dd, J = 9.0, 1.6 Hz, 1H), 7.01 (d, J = 8.4 Hz, 2H), 3.73 (s, 1H), 1.68 (dd, J = 45.1, 15.9 Hz, 5H), 1.38-1.06 (m, 5H); 1313C NMR(75MHz,DMSO)δ165.49,148.84,148.09,143.68,142.71,140.72,138.92,136.99,136.02,132.96,131.15,129.43,124.97,122.81,122.44,120.95,113.78,112.44,109.12,48.59,33.13,25.55;HRMS(+ESI)m / z calcd for C 26 H 25 O3N5[M+H] + 455.1957,found 455.1956.

[0234] Example 24

[0235] 4-((3-(3-Nitrophenyl)-1H-indazol-5-yl)amino)-N-phenylbenzamide (Compound II-2c)

[0236] Step (1): Synthesis of 4-((3-(3-nitrophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)amino)-N-phenylbenzamide (Compound 12c)

[0237] Referring to the method described in Step (2) of Reference Example 19, Compound 7 was replaced with an equimolar amount of Compound 5, and Compound 10c was replaced with an equimolar amount of Compound 10a to obtain Compound 12c (brown oil), which was used directly in the next reaction without purification.

[0238] Step (2): Synthesis of 4-((3-(3-nitrophenyl)-1H-indazol-5-yl)amino)-N-phenylbenzamide (Compound II-2c)

[0239] Referring to the method described in Step (6) of Reference Example 1, Compound 12b was replaced with an equimolar amount of Compound 6a to obtain Compound II-2c, a white solid, and the yield of the two-step reaction was 24%.

[0240] Mp: 280 - 282 °C; 11H NMR(300MHz, DMSO-d6) δ 13.62(s, 1H), 10.03(s, 1H), 8.86 - 8.78(m, 2H), 8.51(d, J = 7.8Hz, 1H), 8.32(ddd, J = 8.2, 2.4, 1.0Hz, 1H), 7.95(dd, J = 12.5, 8.3Hz, 4H), 7.90 - 7.81(m, 2H), 7.75(d, J = 8.9Hz, 1H), 7.49 - 7.38(m, 3H), 7.21 - 7.10(m, 3H); 13 13C NMR(75MHz, DMSO) δ 165.58, 148.91, 140.87, 140.18, 139.15, 136.70, 136.04, 133.08, 131.23, 130.08, 129.08, 124.54, 123.69, 123.10, 122.55, 121.05, 120.79, 113.81, 112.54, 109.87; HRMS(+ESI) m / z calcd for C 26 H 19 O3N5[M + H] + 449.1488, found 449.1485.

[0241] Example 25

[0242] 1. ALK5 Kinase Activity Test and Analysis

[0243] The inhibitory effect of the target compound on ALK5 kinase is reflected by the activity of the kinase treated with the target compound of the present invention, and the inhibitory activity of the compound on the ALK5 interaction is preliminarily evaluated.

[0244] The inhibitory activity of the compounds of the present invention on the ALK5 interaction is shown in Table 1. The results show that the target compounds of the present invention can significantly inhibit the ALK5 interaction.

[0245]

[0246] Table 1. Inhibition Rate of Target Compounds on ALK5 Kinase at a Concentration of 50 nM

[0247]

[0248]

[0249] 2. Cell Proliferation Experiment

[0250] (1), Experimental Purpose: To Determine the Inhibitory Activity of Target Compounds on Human Hepatic Stellate Cells (LX - 2)

[0251] (2) Experimental principle: WST-8 (water-soluble tetrazolium salt: 2-(2-methoxy-4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium monosodium salt) in the CCK-8 (Cell Counting Kit-8) kit can be reduced by certain dehydrogenases in mitochondria under the action of the electron coupling agent 1-methoxy-5-methylphenazinium sulfate dimethyl ester (1-Methoxy PMS) to generate a highly water-soluble orange-yellow formazan dye. The amount of formazan generated is linearly related to the number of live cells, that is, the darker the color, the more live cells, and the weaker the inhibitory ability of the compound on cells.

[0252] (3) Experimental procedure

[0253] Preparation of the drug solution:

[0254] Positive control drug: Add 1 mg of GW-788388 to 50 μL of DMSO to completely dissolve the drug, and prepare a stock solution of 47 μmol / mL for later use. When using, dilute the 47 μmol / mL stock solution 10 times, take 1.36 μl and add it to 100 μL of cell culture medium to make its final concentration 64 μmol / L. Sequentially perform serial dilutions on the 10-fold diluted mother liquor, take 1.36 μL and add it to 100 μL of cell culture medium to make the final drug concentrations 32, 16, 8, 4, 2, 1, 0.5, 0.25 μmol / L respectively.

[0255] Compound I-1b: Add 1 mg of compound I-1b to 50 μL of DMSO to completely dissolve the drug, and prepare a stock solution of 60 μmol / mL for later use. When using, dilute the 60 μmol / mL stock solution 10 times, take 1.07 μL and add it to 100 μL of cell culture medium to make its final concentration 64 μmol / L. Sequentially perform serial dilutions on the 10-fold diluted mother liquor, take 1.07 μL and add it to 100 μL of cell culture medium to make the final drug concentrations 32, 16, 8, 4, 2, 1, 0.5, 0.25 μmol / L respectively.

[0256] Compound II-1a: Add 1 mg of compound II-1a to 50 μL of DMSO to completely dissolve the drug, and prepare a stock solution of 48.4 μmol / mL for later use. When using, dilute the 48.4 μmol / mL stock solution 10 times, take 1.32 μl and add it to 100 μL of cell culture medium to make its final concentration 64 μmol / L. Sequentially perform serial dilutions on the 10-fold diluted mother liquor, take 1.32 μL and add it to 100 μL of cell culture medium to make the final drug concentrations 32, 16, 8, 4, 2, 1, 0.5, 0.25 μmol / L respectively.

[0257] Compound II-1c: 1 mg of compound II-1c was added to 50 μL of DMSO to completely dissolve the drug, and a stock solution of 49.4 μmol / mL was prepared for later use. When in use, the stock solution of 49.4 μmol / mL was diluted 10-fold. 1.30 μL was taken and added to 100 μL of cell culture medium to make the final concentration 64 μmol / L. Subsequently, the 10-fold diluted mother liquor was serially diluted. 1.30 μL was taken and added to 100 μL of cell culture medium to make the final drug concentrations 32, 16, 8, 4, 2, 1, 0.5, and 0.25 μmol / L, respectively.

[0258] Effect of the compound on the activity of LX-2 cells determined by the CCK8 method: LX-2 cells in the logarithmic growth phase were taken, and the cell concentration was adjusted to 1×10 4 cells / ml. They were seeded in 96-well plates at 100 μL / well and cultured in an incubator at 37 °C and 5% CO2 for 24 h. The LX-2 cells were then replaced with complete medium (DMEM + 10% FBS) containing 5 ng / ml TGF-β1 (recombinant human TGF-beta 1 protein (Active), Abcam, (ab50036)) and cultured for another 24 h. Test compounds with different final concentrations (0, 0.25, 0.5, 1, 2, 4, 8, 16, 32, and 64 μmol / L) were added, and three replicates were set for each concentration; GW-788388 (MCE, HY-10326) was used as a positive control drug; the 96-well cell culture plate was placed in the incubator and cultured for another 24 h. 10 μL of CCK8 solution was added to each well and cultured for 2 h. The absorbance value OD450 of each well was measured at 450 nm, the inhibition rate of the drug on the cells was calculated, and the IC 50 value was calculated.

[0259] (4) Experimental results

[0260] Table 2. IC 50

[0261]

[0262] The experimental results are shown in Table 2, indicating that: The IC 50 values of compounds I-1b, II-1a, and II-1c for the LX-2 cell line model with TGF-β-induced fibrotic lesions were approximately 5 times, 4 times, and 17 times that of compound GW-788388, respectively, all showing significant anti-cell proliferation ability and having a therapeutic effect on liver fibrosis.

Claims

1. An indazole compound having a structure as shown in Formula III or a pharmaceutically acceptable salt thereof: Wherein: Ar is selected from 3-pyridyl, 3-nitrophenyl; R is selected from Y is selected from an oxygen atom, a sulfur atom; R 1 selected from hydrogen, fluorine, cyano, formamido, nitro; R 2 Selected from tetrahydropyran-4-yl, cyclohexyl, phenyl.

2. The indazole compound according to claim 1, wherein: Ar is selected from 3-pyridyl, 3-nitrophenyl; Y is selected from an oxygen atom and a sulfur atom; R 1 is selected from hydrogen, o-F, m-F, p-F, m-CN, p-CN, m-CONH2, p-CONH2; R 2 Selected from 3. An indole compound having the following structure or a pharmaceutically acceptable salt thereof:

4. An indazole compound having a structure as shown in Formula III or a pharmaceutically acceptable salt thereof: Wherein: Ar is selected from 3-pyridyl, 3-nitrophenyl; R is selected from Y is an oxygen atom; R 1 selected from o-F, m-CONH2; R 2 selected from Provided that it does not include: Ar is selected from 3-nitrophenyl, and R is selected from R 2 selected from 5. An indazole compound having the following structure or a pharmaceutically acceptable salt thereof: Wherein: Ar is selected from 3-pyridyl; R is selected from Y is an oxygen atom; R 1 selected from o-F; R 2 selected from 6. The indazole compound according to any one of claims 1 to 5, characterized in that: The pharmaceutically acceptable salt is an acid addition salt formed by the indazole compound and the following acids: hydrogen chloride, hydrogen bromide, sulfuric acid, carbonic acid, oxalic acid, citric acid, succinic acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or ferulic acid.

7. Use of the indazole compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5 in the preparation of a drug for treating liver fibrosis diseases.

8. Use of the indazole compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 6 in the preparation of a drug for treating liver fibrosis.

9. A pharmaceutical composition, characterized in that: The pharmaceutical composition uses the indazole compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5 as an active ingredient.

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