5-hydroxy-1H-pyrazole derivative and application thereof

Abstract

The invention discloses a 5-hydroxy-1H-pyrazole derivative and application thereof, and belongs to the technical field of medicines. The 5-hydroxy-1H-pyrazole derivative provided by the invention is novel in structure, has excellent inhibitory activity on IGF2BP2 protein, interferes IGF2BP2-mRNA binding and reduces the stability of related mRNA, and can be used for treating numerous tumor-related diseases, such as breast cancer, colorectal cancer, pancreatic cancer, hepatocellular carcinoma, acute myelogenous leukemia, T-cell acute lymphocytic leukemia, nasopharyngeal carcinoma and the like.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a class of 5-hydroxy-1H-pyrazole derivatives and their uses. Background Technology

[0002] m6A modification is the most common modification in eukaryotic RNA and is currently a hot research topic. Numerous studies have revealed the regulatory role of m6A in cancer development and progression; disruption of m6A homeostasis in vivo often leads to physiological and even pathological changes. RNA m6A modification involves "writers" (m6A methyltransferases), "erasers" (m6A demethylases), and "readers" (m6A methyl recognition proteins). IGF2BP2, as an RNA-binding protein, can specifically recognize m6A modification sites, enhancing the stability and translation of target gene mRNAs (such as MYC, NOTCH1, and SLC1A5), thereby exerting its oncogenic effect. In conclusion, IGF2BP2 can serve as a biomarker for cancer diagnosis and prognosis and has the potential to be a target for various cancer therapies.

[0003] Currently, there are no commercially available small molecule inhibitors of IGF2BP2. Therefore, designing and synthesizing highly active small molecule inhibitors of IGF2BP2 has significant practical application value and is of great importance. Summary of the Invention

[0004] The purpose of this invention is to provide a class of 5-hydroxy-1H-pyrazole derivatives, which can be used as small molecule inhibitors of IGF2BP2 to inhibit RNA readers at the protease level and to treat IGF2BP2-related diseases.

[0005] To achieve the above objectives, the present invention adopts the following technical solution:

[0006] Compounds of general formula I or their pharmaceutically acceptable salts:

[0007]

[0008] R is selected from H, halogen, -COOCH3, carboxyl, hydroxyl, C1-C4 alkyl, C1-C4 alkoxy, amide, trifluoromethyl, nitro or cyano;

[0009] L is selected from amide bonds, methylene, oxyethers, thioethers, ketone carbonyl groups, ester bonds, or C1-C6 saturated carbon chains;

[0010] Ring A is selected from one of the following structures:

[0011]

[0012] Where R aSelected from H, carboxyl, -COOCH3, -CONH2, cyano, hydroxyl, amino, nitro, halogen, trifluoromethyl, C1-C4 alkyl or C1-C4 alkoxy, R b Selected from H, halogen, amino, nitro, hydroxyl, C1-C4 alkyl or C1-C4 alkoxy;

[0013] X and Y may be the same or different from each other, and each is independently selected from carbon, oxygen, nitrogen or sulfur.

[0014] In some preferred embodiments, L is selected from amide bonds, methylene, or oxyethers.

[0015] In some preferred embodiments, ring A is selected from one of the following structures:

[0016]

[0017] Where R a H, hydroxyl, or carboxyl group; R b It can be H, halogen, or methyl;

[0018] X and Y may be the same or different from each other, and each is independently selected from oxygen, nitrogen or sulfur.

[0019] A second objective of this invention is to provide the following compounds or pharmaceutically acceptable salts thereof:

[0020]

[0021]

[0022]

[0023] The compounds of general formula I mentioned above in this invention can also exist in the form of their salts, which are converted into compounds of formula I in vivo. For example, within the scope of this invention, the compounds of this invention are converted into pharmaceutically acceptable salt forms according to processes known in the art, and used in salt form.

[0024] All tautomers of compounds of general formula I of this invention are included within the scope of this invention. The compounds of this invention may exist in specific geometric or stereoisomer forms. Additional asymmetric carbon atoms may be present in alkyl or other substituents; all such isomers and mixtures thereof are included within the scope of this invention.

[0025] The present invention also discloses a method for preparing the relevant compound, such as method a, b or c shown below.

[0026] Method A includes the following steps:

[0027]

[0028] Reagents and conditions: (a) Thioaminourea, 1,4-dioxane, room temperature, 12h; (b) sulfoxide, room temperature, 12h; (c) Miescherichia coli, pyridine, dichloromethane, room temperature, 1h; (d) anhydrous ethanol, 78℃, 3h; (e) acetic acid, 80℃, 2h.

[0029] Method B includes the following steps:

[0030]

[0031] Reagents and conditions: (a) Thioaminourea, 1,4-dioxane, room temperature, 12 h; (f) Diethyl oxaloacetate, acetic acid, 80 °C, 2 h; (g) 4M sodium hydroxide solution, room temperature, 1 h; (h) R-Ph, O-(7-azabenzotriazole)-N,N,N,N-tetramethylurea hexafluorophosphate, N,N-diisopropylethylamine, dichloromethane, room temperature, 12 h.

[0032] Method C includes the following steps:

[0033]

[0034] Reagents and conditions: (e) Acetic acid, 80℃, 2h.

[0035] Method D includes the following steps:

[0036]

[0037] Reagents and conditions: (f) Diethyl oxaloacetate, acetic acid, 80℃, 2h; (g) 4M sodium hydroxide solution, room temperature, 1h; (h) R-Ph, O-(7-azabenzotriazole)-N,N,N,N-tetramethylurea hexafluorophosphate, N,N-diisopropylethylamine, dichloromethane, room temperature, 12h.

[0038] All compounds of general formula I of this invention can be prepared by the methods described above or similarly described, with the appropriate starting materials selected according to the different substituents and their positions. Those skilled in the art should recognize that the above-described route helps in understanding this invention, but does not limit its scope; unless otherwise specified, variables are defined as mentioned in general formula I.

[0039] Another object of the present invention is to provide a pharmaceutical composition comprising an effective amount of an active ingredient and a pharmaceutically acceptable carrier, said active ingredient comprising the above-mentioned compound or a pharmaceutically acceptable salt thereof.

[0040] Another object of the present invention is to provide the use of the above-mentioned compound or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating diseases associated with IGF2BP2 protein dysfunction. These diseases associated with IGF2BP2 protein dysfunction include various tumors such as breast cancer, colorectal cancer, pancreatic cancer, hepatocellular carcinoma, acute myeloid leukemia, T-cell acute lymphoblastic leukemia, and nasopharyngeal carcinoma. Detailed Implementation

[0041] The preparation method of the compound of general formula I of the present invention is described below with reference to specific embodiments, but these specific methods do not constitute any limitation on the present invention. The compounds of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such combinations can be easily performed by those skilled in the art.

[0042] The starting materials and reaction reagents used in the specific embodiments of this invention are all commercially available.

[0043] Experimental methods in the embodiments of this invention that do not specify specific conditions are generally performed under conventional conditions or as recommended by the raw material or product manufacturer. Reagents whose specific source is not specified are commercially available conventional reagents.

[0044] The structure of the compound was determined by 1H NMR using a Bruker Avance 300MHz spectrometer with TMS as an internal standard and a recording temperature of 300K.

[0045] Unless otherwise specified in the examples, all reactions were carried out under an air atmosphere. Unless otherwise specified in the examples, "solution" refers to an aqueous solution. Unless otherwise specified in the examples, the reaction temperature was room temperature, specifically 20–30°C.

[0046] Chemical reactions were detected using a 0.25 mm GF254 thin-layer chromatography silica gel plate and observed using a ZF7 three-way ultraviolet analyzer.

[0047] Example 1: 5-hydroxy-1-(4-(4-hydroxyphenyl)thiazo-2-yl)-N-phenyl-1H-pyrazole-3-carboxamide (1)

[0048]

[0049] Step 1: Synthesize compound b-2

[0050] Compound b-1 (5 g, 23.25 mmol) was dissolved in 20 mL of 1,4-dioxane, and thioaminourea (2.33 g, 25.58 mmol) was added. The mixture was reacted at room temperature for 12 h. The mixture was filtered to obtain a white solid, dried, and then 20 mL of saturated bicarbonate solution was added. The mixture was stirred at room temperature for 4 h, and then filtered to obtain 4.23 g of a yellow solid, with a yield of 87.76%. 1 H NMR(500MHz,Chloroform-d)δ7.95(t,J=3.4Hz,1H),7.66-7.60(m,2H),7.19(s,1H),7. 14(s,1H),6.93-6.87(m,2H),4.56(dd,J=8.3,3.4Hz,1H),4.49(dd,J=8.3,3.4Hz,1H).

[0051] Step 2: Synthesize compound b-3

[0052] Compound b-2 (2 g, 10.63 mmol) was dissolved in 15 mL of acetic acid, and diethyl oxaloacetate (2.64 g, 12.75 mmol) was added. The reaction was carried out at 80 °C for 2 h. After the reaction was completed, the solvent was evaporated under reduced pressure, and sand column chromatography (PE:EA = 10:1) was performed to give 1.41 g of yellow solid, with a yield of 40.06%.

[0053] Step 3: Synthesize compound b-4

[0054] Compound b-3 (1.40 g, 4.41 mmol) was dissolved in 15 mL of water, and 5 mL of 4 M sodium hydroxide solution was added dropwise. The reaction was allowed to proceed at room temperature for 1 h. After the reaction was complete, dilute hydrochloric acid was added dropwise until a solid just precipitated. The mixture was filtered to give 667 mg of a yellow-green solid, with a yield of 49.77%. 1 H NMR (300MHz, DMSO-d6) δ12.82(s,1H),9.57(s,1H),7.68(d,J=8.2Hz,2H),7.62(s,1H),6.73(d,J=8.5Hz,2H),5.84(s,1H).

[0055] Step 4: Synthesize compound 1

[0056] Compound b-4 (200 mg, 659.43 μmol) was dissolved in 10 mL of anhydrous dichloromethane. 2 mL of anhydrous N,N-dimethylformamide was added to aid dissolution. O-(7-azabenzotriazole)-N,N,N,N-tetramethylurea hexafluorophosphate (376.11 mg, 989.15 μmol) and N,N-diisopropylethylamine (127.84 mg, 989.15 μmol) were added, and the reaction was carried out at room temperature for 30 min. Aniline (64.48 mg, 692.40 μmol) was then added, and the reaction was carried out at room temperature for 12 h. After the reaction was complete, the reaction solution was added dropwise to 200 mL of water with vigorous stirring. The mixture was extracted three times with ethyl acetate, and the solution was subjected to sand column chromatography (PE:EA = 5:1) to give 230 mg of a white solid, with a yield of 79.91%. 1 HNMR(300MHz,DMSO-d6)δ10.12(s,1H),9.68(s,1H),7.83(s,2H),7.82-7.79(m,2H),7.78(s,1H),7 .36(t,J=7.8Hz,2H),7.12(t,J=7.3Hz,1H),6.86(dt,J=9.3,2.8Hz,2H),6.09(s,1H),5.77(s,1H).

[0057] Example 2: 5-hydroxy-N-(2-hydroxyphenyl)-1-(4-(4-hydroxyphenyl)thiazolyl-2-yl)-1H-pyrazole-3-carboxamide (2)

[0058]

[0059] The preparation method was the same as in Example 1, except that o-aminophenol was used instead of aniline in the preparation of compound 2. 43 mg of a white solid was obtained, with a yield of 16.53%. 1 H NMR(300MHz,DMSO-d6)δ10.27(s,1H),9.70(s,1H),9.35(s,1H),8.20(s,1H) ,7.78(s,3H),6.94(s,2H),6.85(d,J=8.7Hz,3H),6.05(s,1H),3.17(s,2H).

[0060] Example 3: 5-Hydroxy-N-(4-hydroxyphenyl)-1-(4-(4-hydroxyphenyl)thiazolyl-2-yl)-1H-pyrazole-3-carboxamide

[0061]

[0062] The preparation method was the same as in Example 1, except that aniline was replaced with p-aminophenol in the preparation of compound 3. 37 mg of a white solid was obtained, with a yield of 14.23%. 1H NMR(300MHz,DMSO-d6)δ9.88(s,1H),9.68(s,1H),9.29(s,1H),7.94-7.67(m,3H), 7.56(d,J=8.5Hz,2H), 6.85(d,J=8.3Hz,2H), 6.74(d,J=8.7Hz,2H), 6.03(s,1H).

[0063] Example 4: Methyl 2-(5-hydroxy-1-(4-(4-hydroxyphenyl)thiazo-2-yl)-1H-pyrazole-3-carboxamido)benzoate (4)

[0064]

[0065] The preparation method was the same as in Example 1, except that methyl anthranilate was used instead of aniline in the preparation of compound 4. 165 mg of a white solid was obtained, with a yield of 57.33%. 1 H NMR (300MHz, DMSO-d6) δ12.02 (s, 1H), 9.68 (s, 1H), 8.74 (d, J = 8.4Hz, 1H), 8.12-8.01 (m, 1H), 7.90-7. 77(m,3H),7.69(t,J=7.7Hz,1H),7.24(t,J=7.6Hz,1H),6.90-6.81(m,2H),6.07(s,1H),3.98(s,3H).

[0066] Example 5: Methyl 3-(5-hydroxy-1-(4-(4-hydroxyphenyl)thiazo-2-yl)-1H-pyrazole-3-carboxamido)benzoate (5)

[0067]

[0068] The preparation method was the same as in Example 1, except that methyl m-aminobenzoate was used instead of aniline in the preparation of compound 5. 113 mg of a white solid was obtained, with a yield of 39.26%. 1 H NMR (300MHz, DMSO-d6) δ10.04(s,1H),9.68(s,1H),8.53(s,1H),8.09(d,J=8.3Hz,1H),7.91-7.74(m ,3H),7.71(d,J=7.6Hz,1H),7.51(t,J=7.9Hz,1H),6.85(d,J=8.1Hz,2H),6.08(s,1H),3.88(s,3H).

[0069] Example 6: Methyl 4-(5-hydroxy-1-(4-(4-hydroxyphenyl)thiazo-2-yl)-1H-pyrazole-3-carboxamido)benzoate (6)

[0070]

[0071] The preparation method was the same as in Example 1, except that methyl para-aminobenzoate was used instead of aniline in the preparation of compound 6. 135 mg of a white solid was obtained, with a yield of 46.91%. 1 H NMR (300MHz, DMSO-d6) δ10.46(s,1H),9.67(s,1H),7.98(t,J=6.3Hz,4H),7.81( s,1H),7.78(d,J=3.2Hz,2H),6.84(d,J=8.5Hz,2H),6.08(s,1H),3.83(s,3H).

[0072] Example 7: 2-(5-hydroxy-1-(4-(4-hydroxyphenyl)thiazo-2-yl)-1H-pyrazole-3-carboxamido)benzoic acid (7)

[0073]

[0074] Compound 4 (100 mg, 229.13 μmol) was added to a reaction flask, followed by 15 mL of water and 2 mL of 4M sodium hydroxide solution. The reaction was carried out at room temperature for 1 h. After the reaction was complete, dilute hydrochloric acid was added dropwise until a solid just precipitated. The solid was filtered to obtain 47 mg of a brown solid, with a yield of 48.56%. 1 H NMR (300MHz, DMSO-d6) δ12.47(s,1H),9.73(s,1H),8.79(d,J=8.4Hz,1H),8.08(d,J=7.9Hz,1H),7.8 1(d,J=8.9Hz,3H),7.68(t,J=8.0Hz,1H),7.22(t,J=7.7Hz,1H),6.88(d,J=8.0Hz,2H),6.09(s,1H).

[0075] Example 8: 3-(5-hydroxy-1-(4-(4-hydroxyphenyl)thiazo-2-yl)-1H-pyrazole-3-carboxamido)benzoic acid (8)

[0076]

[0077] The preparation method was the same as in Example 7, except that compound 4 was replaced with compound 5. 78 mg of a brown solid was obtained, with a yield of 67.16%. 1H NMR (300MHz, DMSO-d6) δ10.18(s,1H),8.52(s,1H),8.05(d,J=8.1Hz,1H),7.83(d,J=8. 1Hz,2H),7.68(d,J=7.8Hz,1H),7.58-7.32(m,2H),6.84(d,J=8.1Hz,2H),5.56(s,1H).

[0078] Example 9: 4-(5-hydroxy-1-(4-(4-hydroxyphenyl)thiazo-2-yl)-1H-pyrazole-3-carboxamido)benzoic acid (9)

[0079]

[0080] The preparation method was the same as in Example 7, except that compound 4 was replaced with compound 6. 77 mg of a brown solid was obtained, with a yield of 79.55%. ¹H NMR (300 MHz, DMSO-d6) δ 10.40 (s, 1H), 9.69 (s, 1H), 7.97 (t, J = 6.3 Hz, 4H), 7.83 (d, J = 8.2 Hz, 2H), 7.74 (s, 1H), 6.86 (d, J = 8.2 Hz, 2H), 5.98 (s, 1H).

[0081] Example 10: N-(2-chlorophenyl)-5-hydroxy-1-(4-(4-hydroxyphenyl)thiazolyl-2-yl)-1H-pyrazole-3-carboxamide (10)

[0082]

[0083] The preparation method was the same as in Example 1, except that o-chloroaniline was used instead of aniline in the preparation of compound 10. 24 mg of a white solid was obtained, with a yield of 12.49%. 1 H NMR (300MHz, DMSO-d6) δ9.73 (s, 1H), 9.71 (s, 1H), 8.04 (d, J = 8.1Hz, 1H), 7.87-7.75 (m, 3H), 7.59 (d,J=8.0Hz,1H),7.43(t,J=7.8Hz,1H),7.26(t,J=7.9Hz,1H),6.91-6.83(m,2H),6.09(s,1H).

[0084] Example 11: N-(3-chlorophenyl)-5-hydroxy-1-(4-(4-hydroxyphenyl)thiazo-2-yl)-1H-pyrazole-3-carboxamide (11)

[0085]

[0086] The preparation method was the same as in Example 1, except that m-chloroaniline was used instead of aniline in the preparation of compound 11. 117 mg of a white solid was obtained, with a yield of 42.97%. 1 H NMR(300MHz,DMSO-d6)δ10.35(s,1H),9.69(s,1H),8.01(s,1H),7.90-7.67(m,4H) ,7.39(t,J=8.1Hz,1H),7.18(d,J=8.0Hz,1H),6.86(d,J=8.2Hz,2H),6.09(s,1H).

[0087] Example 12: N-(4-chlorophenyl)-5-hydroxy-1-(4-(4-hydroxyphenyl)thiazolyl-2-yl)-1H-pyrazole-3-carboxamide (12)

[0088]

[0089] The preparation method was the same as in Example 1, except that p-chloroaniline was used instead of aniline in the preparation of compound 12. 167 mg of a white solid was obtained, with a yield of 61.34%. 1 H NMR(300MHz,DMSO-d6)δ10.25(s,1H),9.68(s,1H),7.85(dd,J=16.7,8.3Hz, 4H), 7.74 (s, 1H), 7.42 (d, J = 8.3Hz, 2H), 6.85 (d, J = 8.1Hz, 2H), 5.97 (s, 1H).

[0090] Example 13: 5-hydroxy-1-(4-(4-hydroxyphenyl)thiazolyl-2-yl)-N-(2-methoxyphenyl)-1H-pyrazole-3-carboxamide (13)

[0091]

[0092] The preparation method was the same as in Example 1, except that o-methoxyaniline was used instead of aniline in the preparation of compound 13. 157 mg of a white solid was obtained, with a yield of 58.29%. 1 H NMR (300MHz, DMSO-d6) δ9.71 (s, 1H), 9.37 (s, 1H), 8.26 (d, J = 7.9Hz, 1H), 7.93-7.74 (m, 3H), 7.14 (d,J=4.2Hz,2H),7.01(dt,J=8.6,4.5Hz,1H),6.88(d,J=8.2Hz,2H),6.07(s,1H),3.95(s,3H).

[0093] Example 14: 5-hydroxy-1-(4-(4-hydroxyphenyl)thiazolyl-2-yl)-N-(3-methoxyphenyl)-1H-pyrazole-3-carboxamide (14)

[0094]

[0095] The preparation method was the same as in Example 1, except that m-methoxyaniline was used instead of aniline in the preparation of compound 14. 134 mg of a white solid was obtained, with a yield of 49.75%. 1 H NMR (300MHz, DMSO-d6) δ10.07(s,1H),9.66(s,1H),7.84-7.74(m,3H),7.49(t,J=2.2Hz,1H),7.41(d,J=8.3H z,1H),7.24(t,J=8.1Hz,1H),6.83(d,J=8.3Hz,2H),6.68(dd,J=8.1,2.5Hz,1H),6.06(s,1H),3.74(s,3H).

[0096] Example 15: 5-Hydroxy-1-(4-(4-hydroxyphenyl)thiazolyl-2-yl)-N-(4-methoxyphenyl)-1H-pyrazole-3-carboxamide (15)

[0097]

[0098] The preparation method was the same as in Example 1, except that p-methoxyaniline was used instead of aniline in the preparation of compound 15. 214 mg of a white solid was obtained, with a yield of 79.45%. 1 H NMR(300MHz,DMSO-d6)δ10.01(s,1H),9.67(s,1H),7.84-7.78(m,2H),7.76(s,1H), 7.74-7.67(m,2H),6.96-6.90(m,2H),6.88-6.82(m,2H),6.06(s,1H),3.75(s,3H).

[0099] Example 16: 5-hydroxy-1-(4-(4-hydroxyphenyl)thiazolyl-2-yl)-N-(3-(trifluoromethyl)phenyl)-1H-pyrazole-3-carboxamide (16)

[0100]

[0101] The preparation method was the same as in Example 1, except that m-trifluoromethylaniline was used instead of aniline in the preparation of compound 16. 63 mg of a gray solid was obtained, with a yield of 21.40%. 1H NMR (300MHz, DMSO-d6) δ10.50(s,1H),9.68(s,1H),8.29(s,1H),8.11(d,J=8.2Hz,1H),7.80(d, J=8.2Hz,3H),7.60(t,J=8.0Hz,1H),7.46(d,J=7.9Hz,1H),6.84(d,J=8.2Hz,2H),6.09(s,1H).

[0102] Example 17: 5-hydroxy-1-(4-(4-hydroxyphenyl)thiazolyl-2-yl)-N-(4-(trifluoromethyl)phenyl)-1H-pyrazole-3-carboxamide (17)

[0103]

[0104] The preparation method was the same as in Example 1, except that aniline was replaced with p-trifluoromethylaniline in the preparation of compound 17. 154 mg of a white solid was obtained, with a yield of 52.50%. 1 H NMR (300MHz, DMSO-d6) δ10.53 (s, 1H), 9.70 (s, 1H), 8.08 (d, J = 8.4Hz, 2H), 7.8 3(d,J=8.7Hz,3H),7.75(d,J=8.5Hz,2H),6.87(d,J=8.2Hz,2H),6.12(s,1H).

[0105] Example 18: 3-Benzyl-1-(4-(4-hydroxyphenyl)thiazo-2-yl)-1H-pyrazole-5-ol (18)

[0106]

[0107] Step 1: Synthesize compound b-2

[0108] The preparation method was the same as in Example 1, yielding 4.01 g of dark brown solid, with a yield of 83.19%.

[0109] Step 2: Synthesize compound 18-2

[0110] Compound 18-1 (1 g, 7.34 mmol) was added to a reaction flask, and 5 mL of thionyl chloride was added dropwise using a constant pressure dropping funnel under ice bath conditions. The addition was completed in about 10 min, and the reaction was allowed to proceed under ice bath conditions for 0.5 h. After removing the ice bath, the reaction mixture was allowed to react at room temperature for 12 h. After the reaction was completed, the reaction solution was evaporated to dryness under reduced pressure to obtain a transparent oily substance, which was then added to the next step without purification.

[0111] Step 3: Synthesize compound 18-3

[0112] Michaelis acid (1.17 g, 8.11 mmol) was dissolved in 10 mL of anhydrous dichloromethane, and pyridine (1.28 g, 16.22 mmol) was added. The mixture was stirred in an ice bath for 10 min. Compound 19-2 obtained in the previous step was dissolved in 5 mL of anhydrous dichloromethane, and then added dropwise to the reaction system. After the addition was complete, the mixture was reacted in an ice bath for 0.5 h, and then at room temperature for 1 h after removing the ice bath. After the reaction was complete, 20 mL of anhydrous dichloromethane was added, and the mixture was washed three times with water, washed with saturated ammonium chloride, dried over sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a brown solid, which was added to the next step without purification.

[0113] Step 4: Synthesize compound 18-4

[0114] The compound 18-3 obtained in the previous step was dissolved in 20 ml of anhydrous ethanol and refluxed at 78 °C for 3 h. After the reaction was completed, the solvent was evaporated under reduced pressure, and the mixture was subjected to sand column chromatography (PE:EA = 20:1) to give 640 mg of a pale yellow oil, with a yield of 48.12%.

[0115] Step 5: Synthesize compound 18

[0116] 18-4 (400 mg, 1.94 mmol) was dissolved in 4 mL of acetic acid, and compound b-2 (482.35 mg, 2.33 mmol) was added. The reaction was carried out at 80 °C for 2 h. After the reaction was completed, the solvent was evaporated under reduced pressure, and sand column chromatography (PE:EA = 10:1) was performed to give 443 mg of white solid, with a yield of 65.44%. 1 H NMR (300MHz, DMSO-d6) δ9.64(s,1H),7.81(d,J=8.2Hz,2H),7.50(s,1H),7.35(d,J= 4.4Hz, 4H), 7.27 (d, J = 4.3Hz, 1H), 6.83 (d, J = 8.4Hz, 2H), 5.22 (s, 1H), 3.93 (s, 2H).

[0117] Example 19: 3-(3-hydroxybenzyl)-1-(4-(4-hydroxyphenyl)thiazo-2-yl)-1H-pyrazole-5-ol (19)

[0118]

[0119] Step 1: Synthesize compound b-2

[0120] The preparation method was the same as in Example 1, yielding 4.01 g of dark brown solid, with a yield of 83.19%.

[0121] Step 2: Synthesize compound 19-2

[0122] The preparation method is the same as in Example 18, except that compound 18-1 is replaced by compound 19-1 to obtain a colorless oily substance, which is then introduced into the next step without purification.

[0123] Step 3: Synthesize compound 19-3

[0124] The preparation method is the same as in Example 18, except that compound 18-2 is replaced by compound 19-2, resulting in a brown solid, which is then added to the next step without purification.

[0125] Step 4: Synthesize compound 19-4

[0126] The preparation method was the same as in Example 19, except that compound 18-3 was replaced with compound 19-3, yielding 680 mg of a yellow oily substance with a yield of 46.57%.

[0127] Step 5: Synthesize compound 19

[0128] The preparation method was the same as in Example 19, except that compound 18-4 was replaced with compound 19-4, yielding 242 mg of a pale yellow solid with a yield of 43.91%. 1 H NMR (300MHz, DMSO-d6) δ12.66(s,1H),9.68(s,1H),9.45(s,1H),7.85(d,J=8.1Hz,2H),7.55(s,1H),7.1 7(t,J=7.7Hz,1H),6.87(d,J=8.2Hz,2H),6.77(s,1H),6.69(d,J=8.0Hz,1H),5.27(s,1H),3.87(s,2H).

[0129] Example 20: 3-(4-hydroxybenzyl)-1-(4-(4-hydroxyphenyl)thiazo-2-yl)-1H-pyrazole-5-ol (20)

[0130]

[0131] Step 1: Synthesize compound b-2

[0132] The preparation method was the same as in Example 1, yielding 4.01 g of dark brown solid, with a yield of 83.18%.

[0133] Step 2: Synthesize compound 20-2

[0134] The preparation method is the same as in Example 18, except that compound 18-1 is replaced with compound 20-1 to obtain a colorless oily substance, which is then introduced into the next step without purification.

[0135] Step 3: Synthesize compound 20-3

[0136] The preparation method is the same as in Example 18, except that compound 18-2 is replaced with compound 20-2 to obtain a brown solid, which is then added to the next step without purification.

[0137] Step 4: Synthesize compound 20-4

[0138] The preparation method was the same as in Example 18, except that compound 18-3 was replaced with compound 20-3, yielding 580 mg of a yellow oily substance with a yield of 39.72%.

[0139] Step 5: Synthesize compound 20

[0140] The preparation method was the same as in Example 18, except that compound 18-4 was replaced with compound 20-4, yielding 267 mg of a pale yellow solid with a yield of 48.60%. 1 H NMR (300MHz, DMSO-d6) δ9.60 (s, 1H), 9.30 (s, 1H), 7.78 (d, J = 8.1Hz, 2H), 7.46 (s, 1 H),7.10(d,J=7.9Hz,2H),6.75(dd,J=31.7,8.2Hz,4H),5.14(s,1H),3.76(s,2H).

[0141] Example 21: 1-(4-(4-hydroxyphenyl)thiazolyl-2-yl)-3-(3-methoxybenzyl)-1H-pyrazole-5-ol (21)

[0142]

[0143] Step 1: Synthesize compound b-2

[0144] The preparation method was the same as in Example 1, yielding 4.01 g of dark brown solid, with a yield of 83.18%.

[0145] Step 2: Synthesize compound 21-2

[0146] The preparation method is the same as in Example 18, except that compound 18-1 is replaced with compound 21-1 to obtain a colorless oily substance, which is then introduced into the next step without purification.

[0147] Step 3: Synthesize compound 21-3

[0148] The preparation method is the same as in Example 18, except that compound 18-2 is replaced by compound 21-2, resulting in a brown solid, which is then added to the next step without purification.

[0149] Step 4: Synthesize compound 21-4

[0150] The preparation method was the same as in Example 18, except that compound 18-3 was replaced with compound 21-3, yielding 614 mg of a yellow oily substance with a yield of 45.82%.

[0151] Step 5: Synthesize compound 21

[0152] The preparation method was the same as in Example 18, except that compound 18-4 was replaced with compound 21-4, yielding 281 mg of white solid with a yield of 63.44%. 1 H NMR (300MHz, DMSO-d6) δ12.61(s,1H),9.63(s,1H),7.81(d,J=8.0Hz,2H),7.50(s,1H),7.26(t,J= 7.7Hz,1H),6.92(d,J=8.3Hz,2H),6.83(d,J=8.3Hz,3H),5.23(s,1H),3.89(s,2H),3.75(s,3H).

[0153] Example 22: 1-(4-(4-hydroxyphenyl)thiazolyl-2-yl)-3-(4-methoxybenzyl)-1H-pyrazole-5-ol (22)

[0154]

[0155] Step 1: Synthesize compound b-2

[0156] The preparation method was the same as in Example 1, yielding 4.01 g of dark brown solid, with a yield of 83.18%.

[0157] Step 2: Synthesize compound 22-2

[0158] The preparation method is the same as in Example 18, except that compound 18-1 is replaced with compound 22-1 to obtain a colorless oily substance, which is then introduced into the next step without purification.

[0159] Step 3: Synthesize compound 22-3

[0160] The preparation method is the same as in Example 18, except that compound 18-2 is replaced with compound 22-2, resulting in a brown solid, which is then added to the next step without purification.

[0161] Step 4: Synthesize compound 22-4

[0162] The preparation method was the same as in Example 18, except that compound 18-3 was replaced with compound 22-3, yielding 720 mg of a yellow oily substance with a yield of 53.73%.

[0163] Step 5: Synthesize compound 22

[0164] The preparation method was the same as in Example 18, except that compound 18-4 was replaced with compound 22-4, yielding 348 mg of white solid with a yield of 78.67%. 1H NMR (300MHz, DMSO-d6) δ12.63(s,1H),9.69(s,1H),7.87(d,J=8.1Hz,2H),7.55(s,1H),7.32(t ,J=8.1Hz,2H),7.00-6.93(m,2H),6.92-6.86(m,2H),5.23(s,1H),3.91(s,2H),3.79(s,3H).

[0165] Example 23: 4-(5-hydroxy-1-(4-(4-hydroxyphenyl)thiazolyl-2-yl)-1H-pyrazole-3-yl)methyl)benzonitrile (23)

[0166]

[0167] Step 1: Synthesize compound b-2

[0168] The preparation method was the same as in Example 1, yielding 4.01 g of dark brown solid, with a yield of 83.18%.

[0169] Step 2: Synthesize compound 23-2

[0170] The preparation method is the same as in Example 18, except that compound 18-1 is replaced with compound 23-1 to obtain a colorless oily substance, which is then introduced into the next step without purification.

[0171] Step 3: Synthesize compound 23-3

[0172] The preparation method is the same as in Example 18, except that compound 18-2 is replaced with compound 23-2, resulting in a brown solid, which is then added to the next step without purification.

[0173] Step 4: Synthesize compound 23-4

[0174] The preparation method was the same as in Example 18, except that compound 18-3 was replaced with compound 23-3, yielding 530 mg of a yellow oily substance with a yield of 57.33%.

[0175] Step 5: Synthesize compound 23

[0176] The preparation method was the same as in Example 18, except that compound 18-4 was replaced with compound 23-4, yielding 30 mg of brown solid with a yield of 6.17%. 1 H NMR (300MHz, DMSO-d6) δ9.61 (s, 1H), 7.80 (t, J = 7.5Hz, 4H), 7.63-7.41 (m, 3H), 6.83 (d, J = 8.7Hz, 2H), 5.30 (s, 1H), 4.02 (s, 2H).

[0177] Example 24: 3-(2-fluorobenzyl)-1-(4-(4-hydroxyphenyl)thiazo-2-yl)-1H-pyrazole-5-ol (24)

[0178]

[0179] Step 1: Synthesize compound b-2

[0180] The preparation method was the same as in Example 1, yielding 4.01 g of dark brown solid, with a yield of 83.18%.

[0181] Step 2: Synthesize compound 24-2

[0182] The preparation method is the same as in Example 18, except that compound 18-1 is replaced with compound 24-1 to obtain a colorless oily substance, which is then introduced into the next step without purification.

[0183] Step 3: Synthesize compound 24-3

[0184] The preparation method is the same as in Example 18, except that compound 18-2 is replaced with compound 24-2 to obtain a brown solid, which is then added to the next step without purification.

[0185] Step 4: Synthesize compound 24-4

[0186] The preparation method was the same as in Example 18, except that compound 18-3 was replaced with compound 24-3, yielding 670 mg of a yellow oily substance with a yield of 51.54%.

[0187] Step 5: Synthesize compound 24

[0188] The preparation method was the same as in Example 18, except that compound 18-4 was replaced with compound 24-4, yielding 140 mg of a light pink solid with a yield of 25.94%. 1 H NMR (300MHz, DMSO-d6) δ9.62 (s, 1H), 7.80 (d, J = 8.7Hz, 2H), 7.51 (s, 1H), 7.46 -7.28(m,2H),7.23(d,J=10.9Hz,2H),6.85(s,2H),5.14(s,1H),3.96(s,2H)..

[0189] Example 25: 3-(3-fluorobenzyl)-1-(4-(4-hydroxyphenyl)thiazo-2-yl)-1H-pyrazole-5-ol (25)

[0190]

[0191] Step 1: Synthesize compound b-2

[0192] The preparation method was the same as in Example 1, yielding 4.01 g of dark brown solid, with a yield of 83.18%.

[0193] Step 2: Synthesize compound 25-2

[0194] The preparation method is the same as in Example 18, except that compound 18-1 is replaced with compound 25-1 to obtain a colorless oily substance, which is then introduced into the next step without purification.

[0195] Step 3: Synthesize compound 25-3

[0196] The preparation method is the same as in Example 18, except that compound 18-2 is replaced with compound 25-2 to obtain a brown solid, which is then added to the next step without purification.

[0197] Step 4: Synthesize compound 25-4

[0198] The preparation method was the same as in Example 18, except that compound 18-3 was replaced with compound 25-3, yielding 700 mg of a yellow oily substance with a yield of 53.84%.

[0199] Step 5: Synthesize compound 25

[0200] The preparation method was the same as in Example 18, except that compound 18-4 was replaced with compound 25-4, yielding 200 mg of brown solid with a yield of 30.52%. 1 H NMR(300MHz,DMSO-d6)δ9.62(s,1H),7.81(d,J=6.5Hz,2H),7.50(s,1H),7.45-7.32(m,1H),7 .19(d,J=7.8Hz,2H),7.09(t,J=8.7Hz,1H),6.83(d,J=8.7Hz,2H),5.28(s,1H),3.94(s,2H).

[0201] Example 26: 3-(4-fluorobenzyl)-1-(4-(4-hydroxyphenyl)thiazo-2-yl)-1H-pyrazole-5-ol (26)

[0202]

[0203] Step 1: Synthesize compound b-2

[0204] The preparation method was the same as in Example 1, yielding 4.01 g of dark brown solid, with a yield of 83.18%.

[0205] Step 2: Synthesize compound 26-2

[0206] The preparation method is the same as in Example 18, except that compound 18-1 is replaced with compound 26-1 to obtain a colorless oily substance, which is then introduced into the next step without purification.

[0207] Step 3: Synthesize compound 26-3

[0208] The preparation method is the same as in Example 18, except that compound 18-2 is replaced with compound 26-2 to obtain a brown solid, which is then added to the next step without purification.

[0209] Step 4: Synthesize compound 26-4

[0210] The preparation method was the same as in Example 18, except that compound 18-3 was replaced with compound 26-3, yielding 660 mg of a yellow oily substance with a yield of 57.77%.

[0211] Step 5: Synthesize compound 26

[0212] The preparation method was the same as in Example 18, except that compound 18-4 was replaced with compound 26-4, yielding 268 mg of white solid with a yield of 42.49%. 1 H NMR (300MHz, DMSO-d6) δ12.63(s,1H),9.65(d,J=2.7Hz,1H),7.82(d,J=8.1Hz,2H),7.52( s,1H),7.40(s,2H),7.19(t,J=8.6Hz,2H),6.90-6.81(m,2H),5.23(s,1H),3.93(s,2H).

[0213] Example 27: 1-(4-(4-hydroxyphenyl)thiazolyl-2-yl)-3-(2-(trifluoromethyl)benzyl)-1H-pyrazole-5-ol (27)

[0214]

[0215] Step 1: Synthesize compound b-2

[0216] The preparation method was the same as in Example 1, yielding 4.01 g of dark brown solid, with a yield of 83.18%.

[0217] Step 2: Synthesize compound 27-2

[0218] The preparation method is the same as in Example 18, except that compound 18-1 is replaced with compound 27-1 to obtain a colorless oily substance, which is then introduced into the next step without purification.

[0219] Step 3: Synthesize compound 27-3

[0220] The preparation method is the same as in Example 18, except that compound 18-2 is replaced with compound 27-2 to obtain a brown solid, which is then added to the next step without purification.

[0221] Step 4: Synthesize compound 27-4

[0222] The preparation method was the same as in Example 18, except that compound 18-3 was replaced with compound 27-3, yielding 520 mg of a yellow oily substance with a yield of 39.09%.

[0223] Step 5: Synthesize compound 27

[0224] The preparation method was the same as in Example 18, except that compound 18-4 was replaced with compound 27-4, yielding 140 mg of pink solid with a yield of 22.99%. 1 H NMR (300MHz, DMSO-d6) δ9.63 (s, 1H), 7.78 (dd, J = 13.6, 8.1Hz, 3H), 7.67 (t, J =7.5Hz,1H),7.54(d,J=9.9Hz,3H),6.85(s,2H),5.01(s,1H),4.11(s,2H).

[0225] Example 28: 1-(4-(4-hydroxyphenyl)thiazolyl-2-yl)-3-(3-(trifluoromethyl)benzyl)-1H-pyrazole-5-ol (28)

[0226]

[0227] Step 1: Synthesize compound b-2

[0228] The preparation method was the same as in Example 1, yielding 4.01 g of dark brown solid, with a yield of 83.18%.

[0229] Step 2: Synthesize compound 28-2

[0230] The preparation method is the same as in Example 18, except that compound 18-1 is replaced by compound 28-1 to obtain a colorless oily substance, which is then introduced into the next step without purification.

[0231] Step 3: Synthesize compound 28-3

[0232] The preparation method is the same as in Example 18, except that compound 18-2 is replaced with compound 28-2 to obtain a brown solid, which is then added to the next step without purification.

[0233] Step 4: Synthesize compound 28-4

[0234] The preparation method was the same as in Example 18, except that compound 18-3 was replaced with compound 28-3, yielding 500 mg of a yellow oily substance with a yield of 37.59%.

[0235] Step 5: Synthesize compound 28

[0236] The preparation method was the same as in Example 18, except that compound 18-4 was replaced with compound 28-4, yielding 92 mg of white solid with a yield of 15.11%.1 H NMR (300MHz, DMSO-d6) δ9.62 (s, 1H), 7.80 (d, J = 8.7Hz, 2H), 7.74 (s, 1H), 7.70 -7.54(m,3H),7.51(s,1H),6.83(d,J=8.7Hz,2H),5.29(s,1H),4.04(s,2H)..

[0237] Example 29: 1-(4-(4-hydroxyphenyl)thiazol-2-yl)-3-(4-(trifluoromethyl)benzyl)-1H-pyrazole-5-ol (29)

[0238]

[0239] Step 1: Synthesize compound b-2

[0240] The preparation method was the same as in Example 1, yielding 4.01 g of dark brown solid, with a yield of 83.18%.

[0241] Step 2: Synthesize compound 29-2

[0242] The preparation method is the same as in Example 18, except that compound 18-1 is replaced with compound 29-1 to obtain a colorless oily substance, which is then introduced into the next step without purification.

[0243] Step 3: Synthesize compound 29-3

[0244] The preparation method is the same as in Example 18, except that compound 18-2 is replaced with compound 29-2 to obtain a brown solid, which is then added to the next step without purification.

[0245] Step 4: Synthesize compound 29-4

[0246] The preparation method was the same as in Example 18, except that compound 18-3 was replaced with compound 29-3, yielding 750 mg of a yellow oily substance with a yield of 56.39%.

[0247] Step 5: Synthesize compound 29

[0248] The preparation method was the same as in Example 18, except that compound 18-4 was replaced with compound 29-4, yielding 167 mg of white solid with a yield of 27.43%. 1 H NMR (300MHz, DMSO-d6) δ9.62 (s, 1H), 7.80 (d, J = 8.7Hz, 2H), 7.74 (s, 1H), 7.70 -7.54(m,3H),7.51(s,1H),6.83(d,J=8.7Hz,2H),5.30(s,1H),4.04(s,2H)..

[0249] Example 30: 1-(4-(4-hydroxyphenyl)thiazolyl-2-yl)-3-(2-methylbenzyl)-1H-pyrazole-5-ol (30)

[0250]

[0251] Step 1: Synthesize compound b-2

[0252] The preparation method was the same as in Example 1, yielding 4.01 g of dark brown solid, with a yield of 83.18%.

[0253] Step 2: Synthesize compound 30-2

[0254] The preparation method is the same as in Example 18, except that compound 18-1 is replaced with compound 30-1 to obtain a colorless oily substance, which is then introduced into the next step without purification.

[0255] Step 3: Synthesize compound 30-3

[0256] The preparation method is the same as in Example 18, except that compound 18-2 is replaced with compound 30-2 to obtain a brown solid, which is then added to the next step without purification.

[0257] Step 4: Synthesize compound 30-4

[0258] The preparation method was the same as in Example 18, except that compound 18-3 was replaced with compound 30-3, yielding 510 mg of a colorless oil with a yield of 35.42%.

[0259] Step 5: Synthesize compound 30

[0260] The preparation method was the same as in Example 18, except that compound 18-4 was replaced with compound 30-4, yielding 212 mg of a pale purple solid with a yield of 32.12%. 1 H NMR (300MHz, DMSO-d6) δ9.62(s,1H),7.81(d,J=7.7Hz,2H),7.50(s,1H),7.21(d,J=14.9Hz,4H),6.83(d,J=8.7Hz,2H),5.00(s,1H),3.92(s,2H).

[0261] Example 31: 1-(4-(4-hydroxyphenyl)thiazolyl-2-yl)-3-(3-methylbenzyl)-1H-pyrazole-5-ol (31)

[0262]

[0263] Step 1: Synthesize compound b-2

[0264] The preparation method was the same as in Example 1, yielding 4.01 g of dark brown solid, with a yield of 83.18%.

[0265] Step 2: Synthesize compound 31-2

[0266] The preparation method is the same as in Example 18, except that compound 18-1 is replaced with compound 31-1 to obtain a colorless oily substance, which is then introduced into the next step without purification.

[0267] Step 3: Synthesize compound 31-3

[0268] The preparation method is the same as in Example 18, except that compound 18-2 is replaced with compound 31-2 to obtain a brown solid, which is then added to the next step without purification.

[0269] Step 4: Synthesize compound 31-4

[0270] The preparation method was the same as in Example 18, except that compound 18-3 was replaced with compound 31-3, yielding 700 mg of a colorless oil with a yield of 48.61%.

[0271] Step 5: Synthesize compound 31

[0272] The preparation method was the same as in Example 18, except that compound 18-4 was replaced with compound 31-4, yielding 95 mg of gray solid with a yield of 14.41%. 1 H NMR (300MHz, DMSO-d6) δ9.62 (s, 1H), 7.81 (d, J = 8.4Hz, 2H), 7.49 (s, 1H), 7.22 (t, J = 7.4Hz, 1H),7.14(s,2H),7.06(d,J=7.8Hz,1H),6.83(d,J=8.6Hz,2H),5.21(s,1H),3.88(s,2H).

[0273] Example 32: 1-(4-(4-hydroxyphenyl)thiazolyl-2-yl)-3-(4-methylbenzyl)-1H-pyrazole-5-ol (32)

[0274]

[0275] Step 1: Synthesize compound b-2

[0276] The preparation method was the same as in Example 1, yielding 4.01 g of dark brown solid, with a yield of 83.18%.

[0277] Step 2: Synthesize compound 32-2

[0278] The preparation method is the same as in Example 18, except that compound 18-1 is replaced with compound 32-1 to obtain a colorless oily substance, which is then introduced into the next step without purification.

[0279] Step 3: Synthesize compound 32-3

[0280] The preparation method is the same as in Example 18, except that compound 18-2 is replaced with compound 32-2, resulting in a brown solid, which is then added to the next step without purification.

[0281] Step 4: Synthesize compound 32-4

[0282] The preparation method is the same as in Example 18, except that compound 18-3 is replaced with compound 32-3, yielding 600 mg of a colorless oil with a yield of 41.67%.

[0283] Step 5: Synthesize compound 32

[0284] The preparation method was the same as in Example 18, except that compound 18-4 was replaced with compound 32-4, yielding 62 mg of brownish-yellow solid with a yield of 9.41%. 1 H NMR(300MHz, DMSO-d6)δ9.62(s,1H),7.81(d,J=8.3Hz,2H),7.49(s,1H),7.22(d,J= 7.8Hz,2H),7.14(d,J=7.9Hz,2H),6.83(d,J=8.3Hz,2H),5.19(s,1H),3.86(s,2H).

[0285] Example 33: 1-(4-(4-hydroxyphenyl)thiazolyl-2-yl)-3-(2-nitrobenzyl)-1H-pyrazole-5-ol (33)

[0286]

[0287] Step 1: Synthesize compound b-2

[0288] The preparation method was the same as in Example 1, yielding 4.01 g of dark brown solid, with a yield of 83.18%.

[0289] Step 2: Synthesize compound 33-2

[0290] The preparation method is the same as in Example 18, except that compound 18-1 is replaced with compound 33-1 to obtain a colorless oily substance, which is then introduced into the next step without purification.

[0291] Step 3: Synthesize compound 33-3

[0292] The preparation method is the same as in Example 18, except that compound 18-2 is replaced with compound 33-2, resulting in a brown solid, which is then added to the next step without purification.

[0293] Step 4: Synthesize compound 33-4

[0294] The preparation method was the same as in Example 18, except that compound 18-3 was replaced with compound 33-3, yielding 330 mg of a yellow oily substance with a yield of 24.44%.

[0295] Step 5: Synthesize compound 33

[0296] The preparation method was the same as in Example 18, except that compound 18-4 was replaced with compound 33-4, yielding 40 mg of a dark red solid with a yield of 8.51%. 1 H NMR (300MHz, DMSO-d6) δ9.62(s,1H),8.05(d,J=8.1Hz,1H),7.84-7.66(m,3H),7.65-7.43(m,3H),6.83(d,J=8.1Hz,2H),4.25(s,2H).

[0297] Example 34: 1-(4-(4-hydroxyphenyl)thiazolyl-2-yl)-3-(3-nitrobenzyl)-1H-pyrazole-5-ol (34)

[0298]

[0299] Step 1: Synthesize compound b-2

[0300] The preparation method was the same as in Example 1, yielding 4.01 g of dark brown solid, with a yield of 83.18%.

[0301] Step 2: Synthesize compound 34-2

[0302] The preparation method is the same as in Example 18, except that compound 18-1 is replaced with compound 34-1 to obtain a colorless oily substance, which is then introduced into the next step without purification.

[0303] Step 3: Synthesize compound 34-3

[0304] The preparation method is the same as in Example 18, except that compound 18-2 is replaced with compound 34-2, resulting in a brown solid, which is then added to the next step without purification.

[0305] Step 4: Synthesize compound 34-4

[0306] The preparation method was the same as in Example 18, except that compound 18-3 was replaced with compound 34-3, yielding 460 mg of a yellow oily substance with a yield of 34.07%.

[0307] Step 5: Synthesize compound 34

[0308] The preparation method was the same as in Example 18, except that compound 18-4 was replaced with compound 34-4, yielding 100 mg of brown solid with a yield of 15.94%. 1H NMR (300MHz, DMSO-d6) δ9.64(s,1H),8.36-8.03(m,2H),7.80(s,3H),7.64(s,1H),7.51(s,1H),6.82(d,J=8.0Hz,2H),5.35(s,1H),4.09(s,2H).

[0309] Example 35: 1-(4-(4-hydroxyphenyl)thiazolyl-2-yl)-3-(4-nitrobenzyl)-1H-pyrazole-5-ol (35)

[0310]

[0311] Step 1: Synthesize compound b-2

[0312] The preparation method was the same as in Example 1, yielding 4.01 g of dark brown solid, with a yield of 83.18%.

[0313] Step 2: Synthesize compound 35-2

[0314] The preparation method is the same as in Example 18, except that compound 18-1 is replaced with compound 35-1 to obtain a colorless oily substance, which is then introduced into the next step without purification.

[0315] Step 3: Synthesize compound 35-3

[0316] The preparation method is the same as in Example 18, except that compound 18-2 is replaced with compound 35-2 to obtain a brown solid, which is then added to the next step without purification.

[0317] Step 4: Synthesize compound 35-4

[0318] The preparation method was the same as in Example 18, except that compound 18-3 was replaced with compound 35-3, yielding 390 mg of a yellow oily substance with a yield of 28.89%.

[0319] Step 5: Synthesize compound 35

[0320] The preparation method was the same as in Example 18, except that compound 18-4 was replaced with compound 35-4, yielding 178 mg of white solid with a yield of 28.63%. 1 H NMR(300MHz,DMSO-d6)δ9.64(s,1H),8.06(d,J=8.3Hz,1H),7.80(d,J=7.9Hz,3H), 7.67-7.57(m,2H),7.53(s,1H),6.83(d,J=8.4Hz,2H),5.09(s,1H),4.25(s,2H).

[0321] Example 36: 3-(2-(5-hydroxy-3-(3-(trifluoromethyl)benzyl)-1H-pyrazol-1-yl)thiazolyl-4-yl)benzoic acid (36)

[0322]

[0323] Step 1: Synthesize compound c-2

[0324] Compound c-1 (1 g, 4.11 mmol) was dissolved in 10 mL of 1,4-dioxane, and thiourea (412.44 mg, 4.53 mmol) was added. The mixture was reacted at room temperature for 12 h. The mixture was filtered to obtain a white solid, dried, and then 20 mL of saturated bicarbonate solution was added. The mixture was stirred at room temperature for 4 h, the pH was adjusted to 4-5 with dilute hydrochloric acid, and the mixture was extracted with ethyl acetate. The solvent was evaporated under reduced pressure to obtain 830 mg of a purple solid, with a yield of 85.75%.

[0325] Step 2: Synthesize compound 28-2

[0326] The preparation method is the same as in Example 28.

[0327] Step 3: Synthesize compound 28-3

[0328] The preparation method is the same as in Example 28.

[0329] Step 4: Synthesize compound 28-4

[0330] The preparation method is the same as in Example 28.

[0331] Step 5: Synthesize compound 36

[0332] The preparation method was the same as in Example 28, except that compound c-2 was used instead of compound b-2, yielding 248 mg of gray solid with a yield of 38.21%. 1 H NMR (300MHz, DMSO-d6) δ8.61-8.45(m,1H),8.02(d,J=7.7Hz,1H),7.85(d,J=7.7Hz ,1H),7.62(d,J=2.8Hz,2H),7.53(s,3H),7.40(s,2H),4.41(s,1H),3.81(s,2H).

[0333] Example 37: 1-(benzo[d]thiazol-2-yl)-3-(3-(trifluoromethyl)benzyl)-1H-pyrazole-5-ol (37)

[0334]

[0335] Step 1: Synthesize compound 28-2

[0336] The preparation method is the same as in Example 28.

[0337] Step 2: Synthesize compound 28-3

[0338] The preparation method is the same as in Example 28.

[0339] Step 3: Synthesize compound 28-4

[0340] The preparation method is the same as in Example 28.

[0341] Step 4: Synthesize compound 37

[0342] The preparation method was the same as in Example 28, except that compound b-2 was replaced with 2-hydrazinobenzo[d]thiazole, yielding 430 mg of white solid with a yield of 78.53%. 1 H NMR (300MHz, Chloroform-d) δ7.83(t,J=8.2Hz,2H),7.57(s,1H),7.55-7.41(m,4H),7.36(t,J=7.7Hz,1H),5.43(s,1H),4.00(s,2H).

[0343] Example 38: 1-(benzo[d]thiazol-2-yl)-3-(4-(trifluoromethyl)benzyl)-1H-pyrazole-5-ol (38)

[0344]

[0345] Step 1: Synthesize compound 29-2

[0346] The preparation method is the same as in Example 29.

[0347] Step 2: Synthesize compound 29-3

[0348] The preparation method is the same as in Example 29.

[0349] Step 3: Synthesize compound 29-4

[0350] The preparation method is the same as in Example 29.

[0351] Step 4: Synthesize compound 38

[0352] The preparation method was the same as in Example 29, except that compound b-2 was replaced with 2-hydrazinobenzo[d]thiazole, yielding 206 mg of white solid with a yield of 78.53%. 1H NMR(300MHz,DMSO-d6)δ8.02(d,J=7.8Hz,1H),7.83(d,J=8.1Hz,1H),7.75(s,1H),7.71 -7.53(m,3H),7.47(t,J=7.6Hz,1H),7.34(t,J=7.6Hz,1H),5.26(s,1H),4.01(s,2H)..

[0353] Example 39: 1-(pyridin-2-yl)-3-(3-(trifluoromethyl)benzyl)-1H-pyrazole-5-ol (39)

[0354]

[0355] Step 1: Synthesize compound 28-2

[0356] The preparation method is the same as in Example 28.

[0357] Step 2: Synthesize compound 28-3

[0358] The preparation method is the same as in Example 28.

[0359] Step 3: Synthesize compound 28-4

[0360] The preparation method is the same as in Example 28.

[0361] Step 4: Synthesize compound 39

[0362] The preparation method was the same as in Example 28, except that compound b-2 was replaced with 2-hydrazinopyridine, yielding 280 mg of a white solid with a yield of 60.12%. 1 H NMR (300MHz, Chloroform-d) δ7.83(t,J=8.2Hz,2H),7.57(s,1H),7.55-7.41(m,4H),7.36(t,J=7.7Hz,1H),5.43(s,1H),4.00(s,2H).

[0363] Example 40: 1-(pyridin-2-yl)-3-(4-(trifluoromethyl)benzyl)-1H-pyrazole-5-ol (40)

[0364]

[0365] Step 1: Synthesize compound 29-2

[0366] The preparation method is the same as in Example 29.

[0367] Step 2: Synthesize compound 29-3

[0368] The preparation method is the same as in Example 29.

[0369] Step 3: Synthesize compound 29-4

[0370] The preparation method is the same as in Example 29.

[0371] Step 4: Synthesize compound 40

[0372] The preparation method was the same as in Example 29, except that compound b-2 was replaced with 2-hydrazinobenzo[d]thiazole, yielding 110 mg of a pale yellow solid with a yield of 23.60%. 1 H NMR(300MHz,Chloroform-d)δ8.25(d,J=5.1Hz,1H),7.98-7.81(m,2H),7.55(d,J=8.0 Hz, 2H), 7.41 (d, J = 8.0 Hz, 2H), 7.15 (dd, J = 7.1, 4.9 Hz, 1H), 5.37 (s, 1H), 3.98 (s, 2H).

[0373] Example 41: 1-(benzo[d]thiazol-2-yl)-5-hydroxy-N-(4-(trifluoromethyl)phenyl)-1H-pyrazole-3-carboxamide (41)

[0374]

[0375] Step 1: Synthesize compound d-1

[0376] 2-Hydroxybenzo[d]thiazole (482.88 mg, 2.92 mmol) was dissolved in 6 mL of acetic acid, and diethyl oxaloacetate (500 mg, 2.66 mmol) was added. The reaction was carried out at 80 °C for 2 h. After the reaction was completed, the solvent was evaporated under reduced pressure, and sand column chromatography (PE:EA = 20:1 to 5:1) was performed to give 312 mg of a yellow solid, with a yield of 35.41%.

[0377] Step 2: Synthesize compound d-2

[0378] Compound c-1 (310 mg, 1.07 mmol) was dissolved in 15 mL of water, and 3 mL of 4 M sodium hydroxide solution was added dropwise. The reaction was allowed to proceed at room temperature for 1 h. After the reaction was complete, dilute hydrochloric acid was added dropwise until a solid just precipitated. The mixture was filtered to give 206 mg of a yellow-green solid, with a yield of 66.45%.

[0379] Step 3: Synthesize compound 41

[0380] Compound d-1 (200 mg, 0.765 mmol) was dissolved in 6 mL of anhydrous dichloromethane. 2 mL of anhydrous N,N-dimethylformamide was added to aid dissolution. O-(7-azabenzotriazole)-N,N,N,N-tetramethylurea hexafluorophosphate (436.52 mg, 1.148 mmol) and N,N-diisopropylethylamine (148.38 mg, 1.148 mmol) were added, and the reaction was carried out at room temperature for 30 min. Then, p-trifluoromethylaniline (147.92 mg, 0.919 mmol) was added, and the reaction was carried out at room temperature for 12 h. After the reaction was complete, the reaction solution was added dropwise to 200 mL of water with vigorous stirring. The mixture was extracted three times with ethyl acetate, and the solution was subjected to sand column chromatography (PE:EA = 5:1) to give 23 mg of a gray solid, with a yield of 7.43%. 1 H NMR (300MHz, DMSO-d6) δ10.57(s,1H),8.13(d,J=7.9Hz,1H),8.07(d,J=8.4Hz,2H),7.98(d ,J=8.0Hz,1H),7.74(d,J=8.5Hz,2H),7.61-7.51(m,1H),7.51-7.41(m,1H),6.12(s,1H).

[0381] Example 42: 5-hydroxy-1-(pyridin-2-yl)-N-(4-(trifluoromethyl)phenyl)-1H-pyrazole-3-carboxamide (42)

[0382]

[0383] Step 1: Synthesize compound e-1

[0384] The preparation method was the same as in Example 41, except that 2-hydrazinopyridine was used instead of 2-hydrazinobenzo[d]thiazole to obtain 370 mg of white solid, with a yield of 59.71%.

[0385] Step 2: Synthesize compound e-2

[0386] The preparation method was the same as in Example 41, except that compound c-1 was replaced with compound d-1, yielding 315 mg of yellow solid with a yield of 96.77%.

[0387] Step 3: Synthesize compound 42

[0388] The preparation method was the same as in Example 41, except that compound e-2 was used instead of compound d-2, yielding 90 mg of white solid with a yield of 17.67%. 1H NMR(300MHz,Chloroform-d)δ8.85(s,1H),8.37(d,J=5.1Hz,1H),8.08-7.93(m,2H),7.83 (d,J=8.4Hz,2H),7.62(d,J=8.3Hz,2H),7.32(ddd,J=6.8,5.1,1.7Hz,1H),6.22(s,1H).

[0389] Example 43: 1-(benzo[d]oxazol-2-yl)-5-hydroxy-N-(4-(trifluoromethyl)phenyl)-1H-pyrazole-3-carboxamide (43)

[0390]

[0391] Step 1: Synthesize compound f-1

[0392] The preparation method was the same as in Example 41, except that 2-hydrazinobenzo[d]thiazole was replaced with 2-hydrazinobenzo[d]thiazole, yielding 320 mg of brown oily substance with a yield of 34.93%.

[0393] Step 2: Synthesize compound f-2

[0394] The preparation method was the same as in Example 41, except that compound f-1 was used instead of compound d-1, yielding 124 mg of a grayish-white solid with a yield of 41.81%.

[0395] Step 3: Synthesize compound 43

[0396] The preparation method was the same as in Example 41, except that compound f-2 was used instead of compound d-2, yielding 35 mg of gray solid with a yield of 18.42%. 1 H NMR (300MHz, DMSO-d6) δ10.65(s,1H),8.08(d,J=8.4Hz,2H),7.89-7.80(m,2H),7.73(d,J=8.4Hz,2H),7.51-7.43(m,2H),6.10(s,1H).

[0397] Example 44: 1-(benzo[d]oxazol-2-yl)-3-(4-(trifluoromethyl)benzyl)-1H-pyrazole-5-ol (44)

[0398]

[0399] Step 1: Synthesize compound 29-2

[0400] The preparation method is the same as in Example 29.

[0401] Step 2: Synthesize compound 29-3

[0402] The preparation method is the same as in Example 29.

[0403] Step 3: Synthesize compound 29-4

[0404] The preparation method is the same as in Example 29.

[0405] Step 4: Synthesize compound 44

[0406] The preparation method was the same as in Example 29, except that compound b-2 was replaced with 2-hydrazinobenzo[d]oxazole, yielding 25 mg of gray solid with a yield of 4.77%. 1 H NMR (300MHz, DMSO-d6) δ7.72 (dd, J=14.4, 6.3Hz, 4H), 7.55 (d, J=8.0Hz, 2H), 7.46-7.30 (m, 2H), 5.47 (s, 1H), 3.98 (s, 2H).

[0407] The following method is used in this invention to express and purify IGF2BP2 protein.

[0408] Escherichia coli BL21(DE3) strain was transfected with a recombinant plasmid, and the cells were revived in sterile LB medium at 37°C. Single colonies were picked and transferred to 10 mL of LB liquid medium (containing 50 μg / mL kanamycin, Amp), and cultured overnight at 37°C with shaking (220 rpm). The culture was then transferred to 1 L of LB liquid medium (containing 50 μg / mL kanamycin, Amp), and cultured at 37°C with shaking (220 rpm) for 6-8 hours until the OD600 reached 0.6-0.8. The culture was then cooled to 12°C, and 1 mMIPTG (Merck) was added to induce expression for 16 hours (180 rpm). The cells were then harvested and stored at -80°C for later use.

[0409] Add 4g of the bacterial sludge from the previous step to 40mL of lysis buffer, mix well, then add PMSF (Beyotime), and sonicate for 40 minutes. The lysed mixture is then centrifuged at low temperature and high speed (10000rpm, 20min, 4℃). The supernatant is filtered (0.4μm microporous membrane) and purified using an AKTA pure25 (GE Healthcare, Life Sciences) His column (equilibration buffer: 20mM pH 8.0 Tris-HCl, 300mM NaCl, 10mM imidazole; elution buffer: 20mM pH 8.0 Tris-HCl, 300mM NaCl, 500mM imidazole). 10% SDS-PAGE is used to confirm the molecular weight and purity of the bands. Dialysis is performed overnight (20mM pH 8.0 Tris-HCl, 300mM NaCl). The obtained protein concentration is determined by BCA and stored at -80℃ for later use.

[0410] Activity testing examples:

[0411] The inhibitory activity of the compound against IGF2BP2 was tested using a fluorescence polarization assay (FP experiment).

[0412] The instrument used in this experiment was a SpectraMax Multi-Mode Microplate Reader (Molecular Devices), with excitation and emission wavelengths of 485 nm and 535 nm, respectively. The protein used was IGF2BP2 at a final concentration of 80 nM, and the probe was fluorescently labeled m6A-ssRNA (sequence) at a final concentration of 5 nM. The buffer formulation for the test system was: 0.1 mM sodium tetraborate, 1.6 mM boric acid, pH = 7.8. The 384-well blackboard used in the experiment was manufactured by Corning.

[0413] Experimental steps:

[0414] The final test volume was 60 μL. Compounds of different concentrations (each compound was serially diluted 6-9 times with a two-fold gradient, initial concentration 100 μM) were added to the wells in the following order: 20 μL of each compound, 20 μL of IGF2BP2 protein, and 20 μL of fluorescent probe. A blank control (40 μL buffer + 20 μL fluorescent probe) and a negative control (20 μL buffer + 20 μL IGF2BP2 protein + 20 μL fluorescent probe) were included in each experiment. After loading, the 384-well plate was covered with aluminum foil and incubated on a shaker at room temperature for 0.5 hours. Fluorescence was read using a SpectraMax Paradigm Multi-Mode Microplate Reader at an excitation wavelength of 485 nm and an emission wavelength of 535 nm. The mP value was calculated. The inhibition rate was calculated using the formula: Inhibition rate (%) = [1 - (Pobs - Pmin) / (Pmax - Pmin)] × 100%. The IC50 was calculated using GraphpadPrism 6.0 software. 50 .

[0415] The experimental results are shown in Table 1.

[0416] Table 1. IC50 of the compounds of the present invention against IGF2BP2 protein 50 value

[0417]

[0418] As shown in Table 1, the compounds obtained in this invention have significant inhibitory activity against IGF2BP2 and can be used as small molecule inhibitors of IGF2BP2 protein to inhibit the binding of IGF2BP2 to mRNA.

[0419] In summary, the 5-hydroxy-1H-pyrazole derivative compounds provided by this invention exhibit significant inhibitory activity against IGF2BP2, making them effective IGF2BP2 inhibitors. Therefore, drugs containing these compounds as active ingredients can be used to prepare medications for treating clinical conditions related to IGF2BP2.

[0420] As described above, although specific preferred embodiments have been referenced and the invention has been shown and described, this should not be construed as limiting the invention itself. Various changes in form and detail may be made without departing from the spirit and scope of the invention as defined in the appended claims.

Claims

1. A compound of Formula I: ###0001### or a pharmaceutically acceptable salt thereof. R is selected from H, halogen, -COOCH3, carboxyl, hydroxyl, C1-C4 alkyl, C1-C4 alkoxy, amido, trifluoromethyl, nitro, or cyano; L is selected from an amide bond, methylene, an oxygen ether bond, a sulfur ether bond, a ketone carbonyl, an ester bond, or a C1-C6 saturated carbon chain; Ring A is selected from one of the following structures: ###0002### Where R a Selected from H, carboxyl, -COOCH3, -CONH2, cyano, hydroxyl, amino, nitro, halogen, trifluoromethyl, C1-C4 alkyl or C1-C4 alkoxy, R b Selected from H, halogen, amino, nitro, hydroxyl, C1-C4 alkyl or C1-C4 alkoxy; X and Y are the same or different from each other and are each independently selected from carbon, oxygen, nitrogen, or sulfur.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein, L is selected from an amide bond, methylene, or an oxygen ether bond.

3. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein Ring A is selected from one of the following structures: ###0003### wherein R a is H, hydroxy or carboxy; R b is H, halo or methyl; X and Y are the same or different from each other and are each independently selected from oxygen, nitrogen, or sulfur.

4. A compound of the following formula: ###0004### or a pharmaceutically acceptable salt thereof.

5. A pharmaceutical composition, characterized by, a pharmaceutically effective amount of an active ingredient comprising a compound of any one of claims 1-4 or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

6. Use of a compound of any one of claims 1-4 or a pharmaceutically acceptable salt thereof for the manufacture of an IGF2BP2 inhibitor.

7. Use of a compound of any one of claims 1-4 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition of claim 5 for the manufacture of a medicament for the treatment of a disease associated with a dysfunction of IGF2BP2 protein.

8. The use according to claim 7, wherein the disease associated with a dysfunction of IGF2BP2 protein is breast cancer, colorectal cancer, pancreatic cancer, hepatocellular carcinoma, acute myeloid leukemia, T-cell acute lymphoblastic leukemia, or nasopharyngeal carcinoma.

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