5-[1,2,4-oxadiazol]-6-oxo-pyrazolopyridine derivatives, processes for their preparation and use

Abstract

The present application relates to 5-[1,2,4-oxadiazole]-6-oxygen-pyrazolopyridine derivatives and preparation method and application thereof, and the 5-[1,2,4-oxadiazole]-6-oxygen-pyrazolopyridine derivatives and pharmaceutically acceptable salts thereof, the general formula is as shown in formula (II): wherein, the definition of substituent group is described in the specification. The 5-[1,2,4-oxadiazole]-6-oxygen-pyrazolopyridine derivatives and pharmaceutically acceptable salts thereof of the present application have the functions of inhibiting Hsp110-STAT3 interaction, thereby down-regulating p-STAT3 and c-Myc levels, inhibiting pulmonary arterial endothelial cell proliferation, migration and apoptosis resistance, reducing pulmonary arterial pressure and improving the degree of pulmonary vascular remodeling, and further playing the role of anti-pulmonary arterial hypertension.

Description

Technical Field

[0001] This invention relates to 5-[1,2,4-oxadiazole]-6-oxopyrazolopyridine derivatives and their pharmaceutically usable salts, and also to methods for their preparation and use as medicines, particularly as anti-pulmonary hypertension drugs. Background Technology

[0002] Pulmonary hypertension (PH) is a progressive and fatal cardiovascular disease characterized by the abnormal proliferation and migration of pulmonary vascular cells, leading to irreversible remodeling of pulmonary vessels. This results in elevated pulmonary arterial pressure (PAP) and pulmonary vascular resistance (PVR), ultimately causing right heart failure and death. The current diagnostic criteria for PH are a mean pulmonary arterial pressure (mPAP) > 20 mmHg and pulmonary vascular resistance ≥ 3 Wood units.

[0003] Studies generally agree that elevated PAP and PVR are closely related to pulmonary vascular remodeling caused by abnormal proliferation and migration of pulmonary artery endothelial cells (HPAECs) and pulmonary artery smooth muscle cells (HPASMCs). These cells, represented by HPAECs, HPASMCs, and HLF-1 fibroblasts, exhibit abnormal proliferation and migration. Simultaneously, endothelial / mesenchymal transition, fibrosis, and inflammatory progression promote pulmonary vascular remodeling, leading to thickening of the pulmonary vessel walls, reduction in diameter, increased pulmonary vascular pressure, right ventricular hypertrophy, and ultimately right heart failure and death. Due to the similarity in pathogenesis to cancer, pulmonary hypertension is often referred to as "cancer of the cardiovascular system." Pulmonary vascular remodeling is an intrinsic pathological feature of pulmonary hypertension, and endothelial dysfunction of pulmonary artery endothelial cells plays a crucial role in mediating these changes.

[0004] Currently, treatment strategies for pulmonary hypertension (PAH) utilize the nitric oxide (NO), endothelin (ET), and prostacyclin pathways, such as the endothelin receptor antagonist bosentan, the phosphodiesterase inhibitor sildenafil, and prostacyclin analogs. These treatments focus on vasodilation, providing short-term symptom relief and improving short-term survival, but they do not address the underlying pathology of pulmonary vascular remodeling. Therefore, drug therapy only offers a five-year survival rate. Consequently, there is an urgent need for potential therapeutic targets and drugs to improve vascular remodeling, thereby addressing PAH at its root.

[0005] Heat shock protein 110 (Hsp110, also known as Hsp105 or HSPH1) is a stress-induced chaperone protein that helps cells survive under stress by preventing the aggregation of misfolded or unfolded proteins. Numerous studies have shown that Hsp110 is upregulated in various cancers and promotes cancer cell proliferation and metastasis by stabilizing cancer cell proteins. Mechanistic studies have shown that Hsp110 can enhance the Wnt / β-catenin pathway, particularly the signal transduction and activating transcription factor 3 (STAT3) pathway, thereby regulating tumor growth.

[0006] Prior art application number 201910492351.1 discloses compounds with general structural formula (Ⅰ), wherein R1 or R2 is selected from hydrogen, methyl, trifluoromethyl, aromatic ring or substituted aromatic ring; R3 is selected from C1-C6 aliphatic chain, C3-C6 aliphatic ring or substituted C3-C6 aliphatic ring, aromatic ring or substituted aromatic ring, aromatic heterocycle or substituted aromatic heterocycle; X and Y are independently selected from hydrogen, hydroxyl, amino, halogen, respectively; R4 is selected from hydrogen, the following structural fragments and their pharmaceutically usable salts -NO-NH2 R5 is selected from C1-C6 aliphatic chains, C3-C6 aliphatic rings or substituted C3-C6 aliphatic rings, aromatic rings or substituted aromatic rings, aromatic heterocycles or substituted aromatic heterocycles; Z is selected from CH2, O, N.

[0007]

[0008] The patent discloses the use of compounds of general formula (I) in the preparation of drugs for treating pulmonary fibrosis. However, no one has used them to treat pulmonary hypertension.

[0009] Prior art with application number CN201610908428.5 discloses substituted pyrazolopyridine derivatives of the following formula and their pharmaceutically acceptable salts, isomers, prodrugs and pharmaceutical compositions.

[0010]

[0011] These compounds are stimulators of soluble guanylate cyclase, wherein R1 represents thiophene or a substituted pyridyl group, R2 represents -NH2, -NHCH3, or -NHC(=O)CH(CH3)2, R3 is hydrogen or a C1-C4 alkyl group, and R4 is a C1-C6 alkyl group. The invention also discloses the preparation of these compounds and their use as pharmaceuticals, particularly as drugs for treating cardiovascular diseases, such as pulmonary hypertension. However, their efficacy still needs improvement. Summary of the Invention

[0012] The purpose of this invention is to provide a 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivative, its preparation method, and its application in the preparation of pulmonary hypertension treatment drugs.

[0013] To achieve the above objectives, the technical solution of the present invention is as follows:

[0014] 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and their pharmaceutically usable salts are given by the general formula (II):

[0015]

[0016] Wherein, R1 or R2 is independently selected from C1-C8 straight-chain or branched alkyl, or substituted C1-C8 straight-chain or branched alkyl, aromatic ring, aromatic heterocyclic, substituted aromatic ring or substituted aromatic heterocyclic;

[0017] R3 is selected from hydrogen, aromatic ring, aromatic heterocyclic ring, substituted aromatic ring or substituted aromatic heterocyclic ring;

[0018] The substituents on the substituted C1-C8 straight-chain or branched alkyl groups are selected from: halogen, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, hydroxyl;

[0019] The substituents on the substituted aromatic heterocycle or the substituted aromatic ring are independently selected from: halogen, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy.

[0020] In one preferred embodiment, R1 or R2 is independently selected from C1-C4 straight-chain or branched alkyl groups or substituted C1-C4 straight-chain or branched alkyl groups; the substituent is a halogen.

[0021] In one preferred embodiment, R1 or R2 is independently selected from: monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoroethyl, trifluoropropyl, trifluorobutyl, trichloromethyl, trichloropropyl, trichloroethyl, trichlorobutyl, tribromomethyl, tribromoethyl, tribromopropyl, tribromobutyl, triiodomethyl, triiodoethyl, triiodopropyl, triiodobutyl.

[0022] In one preferred embodiment, the aromatic ring is selected from benzene, naphthalene, anthracene, and phenanthrene.

[0023] In one preferred embodiment, the aromatic heterocycle is selected from pyrrole, furan, thiophene, imidazole, thiazole, oxazole, pyrazole, isoxazole, thiadiazole, oxadiazole, tetrazolium, pyridine, pyrimidine, pyrazine, pyridazine, purine, quinoline, isoquinoline, indole, acridine, carbazole, and bioelectron isosteres of the above-mentioned ring systems.

[0024] In one preferred embodiment, R3 is selected from substituted aromatic rings or substituted heterocyclic aromatic rings;

[0025] The number of substituents on the substituted aromatic ring is arbitrarily 1-6, and the positions of the substituents are arbitrarily 1-6. The substituents are selected from: halogen, amino, haloalkyl, alkyl, hydroxy, hydroxyalkyl, alkoxy.

[0026] The number of substituents on the substituted aromatic heterocycle is arbitrarily 1-5, and the positions of the substituents are arbitrarily 1-5. The substituents are selected from: halogen, amino, haloalkyl, alkyl, hydroxy, hydroxyalkyl, alkoxy.

[0027] In one preferred embodiment, R3 is selected from substituted aromatic rings and substituted aromatic heterocycles; when the number of substituents is 1, the substituents are selected from: halogens, amino groups, halogenated C1-C4 alkyl groups, C1-C4 alkyl groups, hydroxyl groups, and C1-C4 hydroxyalkyl groups.

[0028] In one preferred embodiment, R3 is selected from substituted aromatic rings; when the number of substituents is 2, and the position of the substituents is arbitrary between 2 and 6, the substituents are selected from: halogens, amino groups, halogenated C1-C4 alkyl groups, C1-C4 alkyl groups, hydroxyl groups, and C1-C4 hydroxyalkyl groups.

[0029] In one preferred embodiment, R3 is selected from substituted aromatic rings; when there are 2 substituents, one substituent is located at position 1 and the other substituent is located at positions 2-6, the substituents at positions 2-6 are selected from: halogen, amino, halogenated C1-C4 alkyl, C1-C4 alkyl, hydroxyl, C1-C4 hydroxyalkyl; the substituent at position 1 is selected from: C1-C5 alkeneoxy, C1-C5 alkylene.

[0030] In one preferred embodiment, the substituent at position 1 is selected from: -C(CH3)2O-, -C((CH3)2)CH2O-, -C((CH3)2)CH2CH2O-, -C(CH3)2-, -C((CH3)2)CH2-, -C((CH3)2)CH2CH2-, -CH2CH2CH2-, -CH2CH2-, -CH2-, -CO-, -CH2CO-, -CH2CH2CO-.

[0031] In one preferred embodiment, R3 is a substituted aromatic heterocycle; when the number of substituents is 1, the substituents are selected from: -C(CH3)3, -C((CH3)2)CH2CH3, -CH2CH2CH3, -CH2CH3, -CH3.

[0032] In one preferred embodiment, R3 is selected from hydrogen, the following structural fragments, and pharmaceutically available salts thereof:

[0033]

[0034] In one preferred embodiment, the structure of the 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivative is shown in the table:

[0035]

[0036]

[0037]

[0038]

[0039]

[0040] In one preferred embodiment, the pharmaceutically acceptable salt is selected from: hydrochloride, sulfate, phosphate, perchlorate, methanesulfonate, trifluoromethanesulfonate, formate, acetate, propionate, butyrate, maleate, succinate, trifluoroacetate, succinate, salicylate, DL-aspartate, D-aspartate, L-aspartate, DL-glutamate, D-glutamate, L-glutamate, glycerate, stearate, DL-tartrate. D-Tartrate, L-Tartrate, (±)Mandelate, (R)-(-)Mandelate, (S)-(+)Mandelate, Citrate, Mucilage, Malonate, Benzoate, DL-Malate, (±)Lactate, L-(+)-Lactate, D-(+)-Lactate, Pyrate, D-α-galacturonate, Glycerate, DL-Cysteine, D-Cysteine, L-Cysteine, (4S)-Hydroxy-L-Proline, Cyclopropane-1,1-dicarboxylate, 2,2-methylmalonate, tyrosine salt, proline salt, fumarate, 1-hydroxy-2-naphthylcarboxate, phosphonoacetate, carbonate, bicarbonate, 3-phosphonopropionate, DL-pyroglutamate, D-pyroglutamate, L-pyroglutamate, p-toluenesulfonate, benzenesulfonate, ethanesulfonate, (±)camphorsulfonate, naphthalenesulfonate, 1R-(-)-camphorsulfonate, 1S-(+)-camphorsulfonate, 1, 5-Naphthalene disulfonate, 1,2-ethane disulfonate, 1,3-propane disulfonate, 3-(N-morpholino)propane sulfonate, biphenyl sulfonate, hydroxyethyl sulfonate, 1-hydroxy-2-naphthalene sulfonate, dihydrogen phosphate, potassium hydrogen phosphate, dipotassium phosphate, potassium phosphate, sodium hydrogen phosphate, disodium phosphate, sodium phosphate, sodium dihydrogen phosphate, calcium phosphate, third-generation calcium phosphate, hexafluorophosphate, vinyl phosphate, 2-hydroxyethyl phosphate, and phenyl phosphate.

[0041] In one preferred embodiment, the 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivative has the following structure:

[0042] 2-[3-(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 1);

[0043] 3-[3-(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 2);

[0044] 4-[3-(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 3);

[0045] 3-[3-(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]aniline (compound 4);

[0046] 2-Chloro-6-[3-(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]aniline (compound 5);

[0047] 5-(5-{2-[(4-chlorophenyl)oxy]propyl-2-yl}-1,2,4-oxadiazacyclopentan-3-yl)-3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridine (compound 6);

[0048] 2-[3-(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 7);

[0049] 3-[3-(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 8);

[0050] 4-[3-(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 9);

[0051] 2-Chloro-6-[3-(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]aniline (compound 10);

[0052] 3-[3-(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]aniline (compound 11);

[0053] 5-(5-{2-[(4-chlorophenyl)oxy]propyl-2-yl}-1,2,4-oxadiazacyclopentan-3-yl)-1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridine (compound 12);

[0054] 1-Methyl-6-{[(3-methylphenyl)methyl]oxy}-5-(1,2,4-oxadiazacyclopentan-3-yl)-3-(trifluoromethyl)pyrazolo[3,4-b]pyridine (compound 13);

[0055] 2-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 14);

[0056] 3-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 15);

[0057] 4-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 16);

[0058] 3-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]aniline (compound 17);

[0059] 4-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]aniline (compound 18);

[0060] 2-Chloro-6-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]aniline (compound 19);

[0061] 6-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]-2-fluoroaniline (compound 20);

[0062] 5-(5-{2-[(4-chlorophenyl)oxy]propyl-2-yl}-1,2,4-oxadiazacyclopentan-3-yl)-1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridine (compound 21);

[0063] 1,3-Dimethyl-6-{[(3-methylphenyl)methyl]oxy}-5-{5-[1-(2-methylpropyl-2-yl)pyrazol-4-yl]-1,2,4-oxadiazacyclopentan-3-yl}pyrazolo[3,4-b]pyridine (compound 22).

[0064] Based on the same inventive concept, the present invention also provides a method for preparing the 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivative, comprising the following steps:

[0065] S1. The cyanoamino group on compound I is converted to an amidine group to obtain compound II;

[0066] S2 and compound II react with formic acid substituted with R3 to prepare the 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivative;

[0067] The structure of compound I is:

[0068] The structure of compound II is as follows:

[0069] R1, R2, and R3 are defined as described above.

[0070] Based on the same inventive concept, the present invention also provides a synthetic route for the 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives, as shown below:

[0071]

[0072] R1, R2, and R3 are defined as described above.

[0073] Based on the same inventive concept, the present invention also provides the use of the 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and their pharmaceutically usable salts in the preparation of Hsp110-STAT3 interaction inhibitors.

[0074] Based on the same inventive concept, the present invention also provides the use of the 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and their pharmaceutically usable salts in the preparation of anti-pulmonary hypertension drugs.

[0075] Based on the same inventive concept, the present invention also provides the use of the 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and their pharmaceutically usable salts in the preparation of drugs that downregulate the expression levels of p-STAT3 and c-Myc.

[0076] Based on the same inventive concept, the present invention also provides the use of the 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and their pharmaceutically usable salts in the preparation of medicaments that inhibit pulmonary artery endothelial cell proliferation, migration and / or apoptosis resistance.

[0077] Based on the same inventive concept, the present invention also provides the use of the 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and their pharmaceutically usable salts in the preparation of medicaments for reducing pulmonary artery pressure and / or improving the degree of pulmonary vascular remodeling.

[0078] The 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and their pharmaceutically usable salts of the present invention inhibit Hsp110-STAT3 interaction, thereby downregulating p-STAT3 and c-Myc levels, inhibiting pulmonary artery endothelial cell proliferation, migration and apoptosis resistance; reducing pulmonary artery pressure, improving the degree of pulmonary vascular remodeling, and thus exerting an anti-pulmonary hypertension effect.

[0079] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0080] 1. The compound synthesized in this invention has a novel structure and different mechanisms of action. It is a novel Hsp110-STAT3 interaction inhibitor containing an oxadiazole core structure.

[0081] 2. The compounds synthesized in this invention exert an anti-pulmonary hypertension effect and can be used as clinical drugs for the treatment of related diseases. Attached Figure Description

[0082] Figure 1 The expression level of Hsp110 in human pulmonary artery endothelial cells after 12 hours of hypoxia under normoxic or hypoxic conditions and compound treatment, as described in Example 24.

[0083] Figure 2 The expression level of p-STAT3 in human pulmonary artery endothelial cells after 12 hours of hypoxia under normoxic or hypoxic conditions and compound treatment, as described in Example 24.

[0084] Figure 3 The c-Myc expression level in human pulmonary artery endothelial cells after 12 hours of hypoxia under normoxic or hypoxic conditions and compound treatment, as described in Example 24.

[0085] Figure 4 In Example 25, the binding of Hsp110 and STAT3 in human pulmonary artery endothelial cells was measured under normoxic conditions and after treatment with different concentrations of compounds for 12 hours. The ratio of Hsp110 level to STAT3 level in the IP group was used as a reference. Detailed Implementation

[0086] The present invention will be described in detail below with reference to examples.

[0087] Example 1

[0088] 2-[3-(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (Compound 1)

[0089]

[0090] 1-Ethyl-3-(dimethylaminopropyl)carbodiimide hydrochloride (0.17 g, 0.887 mmol) and 1-hydroxybenzotriazole (0.13 g, 0.962 mmol) were weighed sequentially into a 30 mL microwave-safe tube. 15 mL of DMF was added to dissolve them, followed by the addition of salicylic acid (0.13 g, 0.941 mmol) and stirring at room temperature for 30 min. After stirring at room temperature, 0.3 g of [(Z)-amino(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)methylethylenedimethyl]hydroxylamine (0.774 mmol) was added. The reaction mixture was then transferred to a microwave oven and reacted at 120 °C for 40 min. TLC monitoring showed complete reaction of the starting materials, and the reaction solution was deep yellow. Cool the reaction solution to room temperature, pour it into 50 mL of water, stir at room temperature for 30 min, and the beaker will contain a white suspension. Extract with ethyl acetate three times, 50 mL each time. Combine the organic phases and wash twice with saturated saline solution, 100 mL each time. Then dry the organic phase with 15 g of anhydrous sodium sulfate for 6 h. The dried ethyl acetate phase was filtered and concentrated under reduced pressure to obtain 0.37 g of white solid. Recrystallization was performed using anhydrous ethanol. 20 mL of ethanol was added to a 100 mL round-bottom flask, and the sample was found to be almost insoluble at room temperature, forming a white suspension. After reflux at 80 °C for 15 min, the solid still did not completely dissolve. Another 10 mL of ethanol was added, and reflux was continued for another 15 min. The solution gradually became clear, but a small amount of solid remained undissolved. Another 5 mL of ethanol was added, and reflux was continued for another 10 min, at which point the sample completely dissolved. The reaction flask was left to stand overnight at room temperature, and a white solid precipitated out. Filtering yielded a white filter cake, which, after drying, yielded 0.19 g of white solid, compound 1, with a yield of 50.1%.

[0091] 1 H NMR (500MHz, CDCl3): δ10.62(s,1H),8.67(s,1H),8.21(d,J=7.9Hz,2H),8.07(d,J=7.9Hz,1H),7.56(dd,J=16.5,8.6Hz,3H),7.48-7. 39(m,2H),7.32(m,2H),7.19(d,J=8.3Hz,1H),7.14(d,J=7.5Hz,1H),7.09(t,d,J=7.5Hz,1H),5.70(s,2H),2.68(s,3H),2.38(s,3H).

[0092] 13C NMR (126MHz, CDCl3): δ173.56,165.08,160.64,158.13,149.36,144.14,139.11,138.16,136.61,135.27,133.92,128.98×2 ,128.61,128.39,128.31,127.90,125.75,124.62,120.65×2,120.18,117.77,112.17,108.15,105.14,69.02,21.46,12.66.

[0093] HRMS(ESI)m / z calcd for[C 29 H 23 N5O3+H] + :490.1874; found:490.1876[M+H] + .

[0094] Example 2

[0095] 3-[3-(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 2)

[0096]

[0097] Following the synthetic method of compound 1, compound [(Z)-amino(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)methyl hydroxylamine] and 3-hydroxybenzoic acid were used as raw materials to obtain a white solid, namely compound 2, with a yield of 34.4%.

[0098] 1 H NMR (400MHz, DMSO-d6): δ10.13(s,1H),8.87(s,1H),8.20(d,J=7.9Hz,2H),7.67-7.4 0(m,7H),7.31(m,2H),7.12(d,J=7.9Hz,2H),5.62(s,2H),2.61(s,3H),2.29(s,3H).

[0099] 13C NMR (101MHz, DMSO-d6): δ174.63,166.29,160.37,158.47,148.78,144.57,139.14,137.81,137.12,134.88,134.84,131.19, 129.52,128.65×2,128.17,125.89,124.74,124.65,120.80×2,120.05,119.01,114.60,112.11,105.91,68.76,21.49,12.61.

[0100] HRMS(ESI)m / z calcd for[C 29 H 23 N5O3+H] + :490.1874; found:490.1876[M+H] + .

[0101] Example 3

[0102] 4-[3-(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 3)

[0103]

[0104] Following the synthetic method of compound 1, compound [(Z)-amino(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)methyl hydroxylamine] and 4-hydroxybenzoic acid were used as raw materials to obtain a white solid, namely compound 3, with a yield of 43.8%.

[0105] 1 H NMR (400MHz, DMSO-d6): δ10.65(s,1H),8.88(s,1H),8.21(d,J=7.7Hz,2H),8.07(d,J=8.7Hz,2H),7.58(t,J=8.0 Hz,2H),7.44-7.27(m,4H),7.12(d,J=7.5Hz,1H),7.03(d,J=8.7Hz,2H),5.63(s,2H),2.61(s,3H),2.30(s,3H).

[0106] 13C NMR (101MHz, DMSO-d6): δ174.60,165.97,162.59,160.28,148.61,144.35,139.15,137.72,137.12,134.51,130.37×2, 129.40×2,128.58,128.55,128.06,125.72,124.55,119.91×2,116.69×2,114.42,111.97,106.05,68.63,21.48,12.52.

[0107] HRMS(ESI)m / z calcd for[C 29 H 23 N5O3+H] + :490.1874; found:490.1876[M+H] + .

[0108] Example 4

[0109] 3-[3-(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]aniline (compound 4)

[0110]

[0111] Following the synthetic method of compound 1, compound [(Z)-amino(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)methyl hydroxylamine] and 3-aminobenzoic acid were used as raw materials to obtain a yellow solid, namely compound 4, with a yield of 50.8%.

[0112] 1 H NMR (500MHz, DMSO-d6): δ8.87(s,1H),8.21(d,J=8.0Hz,2H),7.58(t,J=7.8Hz,2H),7.41(dd,J=18.6,10.4Hz ,3H),7.39-7.23(m,4H),7.12(d,J=7.3Hz,1H),6.89(d,J=7.9Hz,1H),5.63(m,4H),2.62(s,3H),2.29(s,3H).

[0113] 13C NMR (126MHz, DMSO-d6): δ175.44,166.33,160.57,150.11,148.92,144.73,139.16,137.86,137.19,135.13,130.49,129.64× 2,128.70,128.69,128.24,126.03,124.69,124.20,120.21×2,118.88,115.41,112.71,112.21,106.21,68.82,21.50,12.71.

[0114] HRMS(ESI)m / z calcd for[C 29 H 24 N6O2+H] + :489.2034; found:489.2040[M+H] + .

[0115] Example 5

[0116] 2-Chloro-6-[3-(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]aniline (Compound 5)

[0117]

[0118] Following the synthetic method of compound 1, compound [(Z)-amino(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)methyl hydroxylamine] and 2-amino-3-chlorobenzoic acid were used as raw materials to obtain a yellow solid, namely compound 5, with a yield of 32.4%.

[0119] 1 H NMR (500MHz, DMSO-d6): δ8.98(s,1H),8.22(d,J=7.8Hz,2H),7.95(d,J=8.0Hz,1H),7.64-7.55(m,3H),7.43-7 .28(m,4H),7.13(d,J=7.5Hz,1H),7.04(s,2H),6.79(t,J=7.9Hz,1H),5.64(s,2H),2.64(s,3H),2.29(s,3H).

[0120] 13C NMR (101MHz, DMSO-d6): δ173.43,165.69,160.66,144.78,144.58,139.20,137.93,137.10,135.52,134.31,129.68×2,128.8 0,128.78,128.52,128.29,126.15,124.98,120.31×2,119.74,116.91,112.28,106.05,105.84,69.01,65.49,21.46,12.79.

[0121] HRMS(ESI)m / z calcd for[C 29 H 23 ClN6O2+H] + :523.1587; found:523.1592[M+H] + .

[0122] Example 6

[0123] 5-(5-{2-[(4-chlorophenyl)oxy]propyl-2-yl}-1,2,4-oxadiazacyclopentan-3-yl)-3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridine (compound 6)

[0124]

[0125] Following the synthetic method of compound 1, compound [(Z)-amino(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)methylylidene]hydroxylamine and 2-[(4-chlorophenyl)oxy]-2-methylpropionic acid were used as starting materials to obtain a white solid, namely compound 6, with a yield of 41.1%.

[0126] 1 H NMR (500MHz, DMSO-d6): δ8.76(s,1H),8.20(d,J=7.7Hz,2H),7.57(t,J=8.0Hz,2H),7.40-7.31(m,3H),7.30 -7.24(m,3H),7.12(d,J=7.5Hz,1H),6.85-6.78(m,3H),5.60(s,2H),2.58(s,3H),2.29(s,3H),1.86(s,6H).

[0127] 13C NMR (126MHz, DMSO-d6): δ179.93,165.89,160.43,153.50,148.86,144.61,139.12,137.81,136.99,135.00,129.82×2,129.53×2,12 8.73,128.64,128.30,128.23,126.00,125.96,124.78,123.34×2,120.12×2,112.06,105.59,76.63,68.86,26.35×2,21.46,12.58.

[0128] HRMS(ESI)m / z calcd for[C 32 H 28 ClN5O3+H] + :566.1953; found:566.1960[M+H] + .

[0129] Example 7

[0130] 2-[3-(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 7)

[0131]

[0132] Following the synthetic method of compound 1, using compound [(Z)-amino(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)methyl hydroxylamine and 2-hydroxybenzoic acid as starting materials, a white solid, namely compound 7, was obtained with a yield of 42.1%.

[0133] 1 H NMR (500MHz, DMSO-d6): δ10.65(s,1H),8.82(s,1H),8.02(dd,J=7.8,1.4Hz,1H),7.57-7.52(m,1H),7.47-7. 40(m,2H),7.29(t,J=7.8Hz,1H),7.14(m,2H),7.04(t,J=7.5Hz,1H),5.62(s,2H),4.12(s,3H),2.32(s,3H).

[0134] 13C NMR (126MHz, DMSO-d6): δ174.59,161.07,157.66,156.99,149.43,137.92,136.60,136.59,135.26,133.61,13 2.84,131.24,130.66,130.62,128.89,128.73,128.63,125.16,120.18,117.88,106.85,68.98,34.89,21.48.

[0135] HRMS(ESI)m / z calcd for[C 24 H 18 F3N5O3+H] + :482.1435; found:482.1438[M+H] + .

[0136] Example 8

[0137] 3-[3-(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 8)

[0138]

[0139] Following the synthetic method of compound 1, using compound [(Z)-amino(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)methyl hydroxylamine and 3-hydroxybenzoic acid as raw materials, a white solid, namely compound 8, was obtained with a yield of 34.2%.

[0140] 1 H NMR (500MHz, DMSO-d6): δ10.10(s,1H),8.76(s,1H),7.61(d,J=7.7Hz,1H),7.58-7.54(m,1H),7. 48-7.41(m,3H),7.29(t,J=7.9Hz,1H),7.17-7.09(m,2H),5.63(s,2H),4.12(s,3H),2.31(s,3H).

[0141] 13C NMR (126MHz, DMSO-d6): δ174.75,165.80,160.89,158.43,149.27,137.86,136.55,133.18,132.62,131.14,12 8.77,128.66,128.40,124.90,124.44,121.66,120.87,118.93,114.47,109.04,106.77,68.91,34.77,21.47.

[0142] HRMS(ESI)m / z calcd for[C 24 H 18 F3N5O3+H] + :482.1435; found:482.1438[M+H] + .

[0143] Example 9

[0144] 4-[3-(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 9)

[0145]

[0146] Following the synthetic method of compound 1, using compound [(Z)-amino(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)methyl hydroxylamine and p-hydroxybenzoic acid as raw materials, a white solid, namely compound 9, was obtained with a yield of 31.6%.

[0147] 1 H NMR (500MHz, DMSO-d6): δ10.59(s,1H),8.78(s,1H),8.05(d,J=8.7Hz,2H),7.45(d,J=7.0Hz,2H), 7.30(t,J=7.9Hz,1H),7.15(d,J=7.6Hz,1H),7.00(m,2H),5.65(s,2H),4.13(s,3H),2.32(s,3H).

[0148] 13C NMR (126MHz, DMSO-d6): δ174.94,165.70,162.52,161.00,149.33,137.88,136.64,133.24,132.47,130.55,12 8.83,128.71,128.51,125.05,125.02,122.76,121.24,116.69,114.30,109.39,106.82,68.91,34.85,21.51.

[0149] HRMS(ESI)m / z calcd for[C 24 H 18 F3N5O3+H] + :482.1435; found:482.1438[M+H] + .

[0150] Example 10

[0151] 2-Chloro-6-[3-(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]aniline (Compound 10)

[0152]

[0153] Following the synthetic method of compound 1, compound [(Z)-amino(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)methyl hydroxylamine and 2-amino-3-chlorobenzoic acid were used as raw materials to obtain a yellowish-brown solid, namely compound 10, with a yield of 36.7%.

[0154] 1 H NMR (500MHz, DMSO-d6): δ8.79(s,1H),7.89(d,J=7.9Hz,1H),7.60(d,J=7.7Hz,1H),7.42(m,2H),7.29(t,J=7 .5Hz,1H,),7.14(d,J=7.4Hz,1H),6.99(s,2H),6.76(t,J=7.9Hz,1H),5.63(s,2H),4.13(s,3H),2.31(s,3H).

[0155] 13C NMR (126MHz, DMSO-d6): δ175.66,165.74,160.98,150.01,149.20,137.78,136.62,133.23,130.54,128.91,1 28.53,124.95,123.98,122.72,120.41,118.98,115.48,112.79,109.44,106.77,99.97,68.72,34.79,21.74.

[0156] HRMS(ESI)m / z calcd for[C 24 H 18 ClF3N6O2+H] + :515.1205; found:515.1208[M+H] + .

[0157] Example 11

[0158] 3-[3-(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]aniline (compound 11)

[0159]

[0160] Following the synthetic method of compound 1, using compound [(Z)-amino(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)methyl hydroxylamine and 3-aminobenzoic acid as raw materials, a yellowish-brown solid, namely compound 11, was obtained with a yield of 36.6%.

[0161] 1 H NMR (500MHz, DMSO-d6): δ8.79(s,1H),7.44(m,3H),7.30(m,3H),7.15(d,J=7 .5Hz,1H),6.88(m,1H),5.66(s,2H),5.62(s,2H),4.14(s,3H),2.32(s,3H).

[0162] 13C NMR (126MHz, DMSO-d6): δ175.57,165.84,161.03,150.10,149.40,137.91,136.62,133.31,132.43,130.46,12 8.83,128.72,128.53,125.01,123.98,121.67,118.93,115.34,112.67,109.34,106.84,68.96,34.88,21.50.

[0163] HRMS(ESI)m / z calcd for[C 24 H 19 F3N6O2+H] + :481.1594; found:481.1601[M+H] + .

[0164] Example 12

[0165] 5-(5-{2-[(4-chlorophenyl)oxy]propyl-2-yl}-1,2,4-oxadiazacyclopentan-3-yl)-1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridine (compound 12)

[0166]

[0167] Following the synthetic method of compound 1, using compound [(Z)-amino(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)methylylidene]hydroxylamine and 2-[(4-chlorophenyl)oxy]-2-methylpropionic acid as starting materials, a white solid, namely compound 12, was obtained with a yield of 45.2%.

[0168] 1 H NMR (500MHz, DMSO-d6): δ8.68(s,1H),7.41(d,J=8.7Hz,2H),7.30-7.23(m,3H),7.14(d, J=7.5Hz,1H),6.79(d,J=8.8Hz,2H),5.64(s,2H),4.14(s,3H),2.31(s,3H),1.84(s,6H).

[0169] 13C NMR (126MHz, DMSO-d6): δ180.24,165.48,160.99,153.43,149.44,137.91,136.47,133.51,132.69,129.78×2,12 8.94,128.72,128.62,128.19,125.13,123.30×2,121.62,108.84,106.73,76.62,69.05,34.88,26.30×2,21.47.

[0170] HRMS(ESI)m / z calcd for[C 27 H 23 ClF3N5O3+H] + :558.1514; found:558.1519[M+H] + .

[0171] Example 13

[0172] 1-Methyl-6-{[(3-methylphenyl)methyl]oxy}-5-(1,2,4-oxadiazacyclopentan-3-yl)-3-(trifluoromethyl)pyrazolo[3,4-b]pyridine

[0173] Following the synthetic method of compound 1, compound [(Z)-amino(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)methyl hydroxylamine and formic acid were used as raw materials to obtain a white solid, namely compound 13, with a yield of 43.9%.

[0174] 1 H NMR (500MHz, DMSO-d6): δ9.78(s,1H),8.73(s,1H),7.44(s,1H),7.40(d,J=7.7Hz,1H) ,7.29(t,J=7.6Hz,1H),7.15(d,J=7.5Hz,1H),5.65(s,2H),4.14(s,3H),2.33(s,3H).

[0175] 13 C NMR (126MHz, DMSO-d6): δ166.96,164.44,160.96,149.38,137.91,136.55,133.44,13 2.52,128.87,128.72,128.65,125.08,121.62,108.95,106.81,68.98,34.87,21.49.

[0176] HRMS(ESI)m / z calcd for[C 18 H 14 F3N5O2+H] + :390.1100; found:390.1108[M+H] + .

[0177] Example 14

[0178] 2-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 14)

[0179]

[0180] Following the synthetic method of compound 1, using [(Z)-amino(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)methyl alkylene]hydroxylamine and 2-hydroxybenzoic acid as raw materials, a white solid, namely compound 14, was obtained with a yield of 59.1%.

[0181] 1 H NMR (500MHz, DMSO-d6): δ10.60(s,1H),8.87(s,1H),8.04(d,J=6.5Hz,1H),7.56(t,J=7.1Hz,1H),7.44(d,J=5.9Hz,2H), 7.29(t,J=7.8Hz,1H),7.14(t,J=7.5Hz,2H),7.07(t,J=7.5Hz,1H),5.61(s,2H),3.96(s,3H),2.52(s,3H),2.31(s,3H).

[0182] 13 C NMR (126MHz, DMSO-d6): δ174.13,165.78,160.49,157.63,149.77,142.15,137.86,137.16,135.32,135.26,1 30.29,128.74,128.70,128.62,125.11,120.27,117.93,110.19,110.03,104.72,68.38,33.66,21.50,12.62.

[0183] HRMS(ESI)m / z calcd for[C 24 H 21 N5O3+H] +:428.1717; found:428.1722[M+H] + .

[0184] Example 15

[0185] 3-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 15)

[0186]

[0187] Following the synthetic method of compound 1, using [(Z)-amino(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)methyl alkyl]hydroxylamine and 3-hydroxybenzoic acid as raw materials, a white solid, namely compound 15, was obtained with a yield of 56.4%.

[0188] 1 H NMR (500MHz, DMSO-d6): δ10.10(s,1H),8.79(s,1H),7.65(d,J=7.7Hz,1H),7.59(s,1H),7.51-7.42(m,3H) ,7.29(t,J=7.5Hz,1H),7.12(dd,J=10.4,4.6Hz,2H),5.61(s,2H),3.95(s,3H),2.52(s,3H),2.32(s,3H).

[0189] 13 C NMR (126MHz, DMSO-d6): δ174.60,166.73,160.41,158.49,149.68,142.04,137.82,137.17,134.91,131.24,1 28.75,128.67,128.48,124.97,124.82,120.81,119.01,114.59,110.08,104.86,68.31,33.61,21.51,12.55.

[0190] HRMS(ESI)m / z calcd for[C 24 H 21 N5O3+H] + :428.1717; found:428.1721[M+H] + .

[0191] Example 16

[0192] 4-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 16)

[0193]

[0194] Following the synthetic method of compound 1, using [(Z)-amino(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)methyl alkylene]hydroxylamine and 4-hydroxybenzoic acid as starting materials, a white solid, namely compound 16, was obtained with a yield of 48.7%.

[0195] 1 H NMR (500MHz, DMSO-d6): δ10.55(s,1H),8.77(s,1H),8.06(d,J=8.7Hz,2H),7.44(d,J=7.3Hz,2H),7.28(t,J= 7.7Hz,1H),7.13(d,J=7.6Hz,1H),7.01(d,J=8.7Hz,2H),5.60(s,2H),3.95(s,3H),2.51(s,3H),2.31(s,3H).

[0196] 13 C NMR (126MHz, DMSO-d6): δ174.65,166.51,162.40,160.40,149.63,141.95,137.79,137.21,134.78,1 30.49,128.75,128.64,128.45,124.94,116.70,114.63,110.04,105.10,68.26,33.59,21.52,12.54.

[0197] HRMS(ESI)m / z calcd for[C 24 H 21 N5O3+H] + :428.1717; found:428.1721[M+H] + .

[0198] Example 17

[0199] 3-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]aniline (compound 17)

[0200]

[0201] Following the synthetic method of compound 1, [(Z)-amino(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)methyl hydroxylamine and 3-aminobenzoic acid were used as raw materials to obtain a yellowish-brown solid, namely compound 17, with a yield of 41.0%.

[0202] 1 H NMR (500MHz, DMSO-d6): δ8.77(s,1H),7.48-7.41(m,3H),7.30(m,3H),7.13(d,J=7. 5Hz,1H),6.88(d,J=8.6Hz,1H),5.60(m,4H),3.95(s,3H),2.51(s,3H),2.31(s,3H).

[0203] 13 C NMR (126MHz, DMSO-d6): δ175.20,166.58,160.36,150.06,149.62,141.95,137.80,137.17,134.70,130.41,1 28.63,128.61,128.40,124.88,124.27,118.79,115.40,112.71,110.02,104.98,68.27,33.56,21.51,12.53.

[0204] HRMS(ESI)m / z calcd for[C 24 H 22 N6O2+H] + :427.1877; found:427.1882[M+H] + .

[0205] Example 18

[0206] 4-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]aniline (compound 18)

[0207]

[0208] Following the synthetic method of compound 1, using [(Z)-amino(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)methyl hydroxylamine and 4-aminobenzoic acid as raw materials, a pale yellow solid, namely compound 18, was obtained with a yield of 46.2%.

[0209] 1 H NMR (400MHz, DMSO-d6): δ8.76(s,1H),7.85(d,J=12.4Hz,2H),7.45(m,2H),7.28(t,J=7.9Hz,1H),7.13(d,J =7.4Hz,1H),6.70(d,J=15.2,8.5Hz,2H),6.21(s,2H),5.59(s,2H),3.95(s,3H),2.51(s,3H),2.31(s,3H).

[0210] 13 C NMR (101MHz, DMSO-d6): δ175.16,166.28,160.42,153.99,149.57,141.91,137.78,137.24,134.71,134 .63,130.10,128.64×2,128.44,124.95,113.89×2,110.03,110.01,105.40,68.21,33.60,21.55,12.57.

[0211] HRMS(ESI)m / z calcd for[C 24 H 22 N6O2+H] + :427.1877; found:427.1883[M+H] + .

[0212] Example 19

[0213] 2-Chloro-6-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]aniline (Compound 19)

[0214]

[0215] Following the synthetic method of compound 1, [(Z)-amino(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)methyl hydroxylamine and 2-amino-3-chlorobenzoic acid were used as raw materials to obtain a yellowish-brown solid, namely compound 19, with a yield of 31.6%.

[0216] 1H NMR (500MHz, DMSO-d6): δ8.85(s,1H),7.92(d,J=8.0Hz,1H),7.61(dd,J=7.8,1.2Hz 1H),7.42(m,2H),7.29(t,J=7.5Hz,1H),7.14(d,J=7.5Hz,1H),7.01(s,2H) ,6.78(t,J=7.9Hz,1H),5.60(s,2H),3.95(s,3H),2.52(s,3H),2.31(s,3H).

[0217] 13 C NMR (126MHz, DMSO-d6): δ173.22,165.90,160.42,149.69,144.53,142.06,137.87,137.08,135.16,135.08,1 34.23,134.20,128.73,128.22,125.22,119.70,116.80,110.11,106.08,104.55,68.44,33.63,21.47,12.62.

[0218] HRMS(ESI)m / z calcd for[C 24 H 21 ClN6O2+H] + :461.1487; found:461.1490[M+H] + .

[0219] Example 20

[0220] 6-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]-2-fluoroaniline

[0221]

[0222] Following the synthetic method of compound 1, using (Z)-amino(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)methyl hydroxylamine and 2-amino-3-fluorobenzoic acid as raw materials, a pale yellow solid, namely compound 20, was obtained with a yield of 34.2%.

[0223] 1H NMR (500MHz, DMSO-d6): δ8.89(s,1H),7.74(d,J=8.1Hz,1H),7.42(d,J=8.0Hz,2H),7.39-7.33(m,1H),7.29(t,J= 7.5Hz,1H),7.13(d,J=7.5Hz,1H),6.88(s,2H),6.73(m,1H),5.58(s,2H),3.95(s,3H),2.52(s,3H),2.32(s,3H).

[0224] 13 C NMR (126MHz, DMSO-d6): δ173.19,165.99,160.42,152.34,150.43,149.67,142.09,137.99,137.85,137.11,1 35.29,128.72,128.66,125.14,124.61,119.00,115.63,110.13,106.48,104.65,68.38,33.62,21.48,12.64.

[0225] HRMS(ESI)m / z calcd for[C 24 H 21 FN6O2+H] + :445.1783; found:445.1784[M+H] + .

[0226] Example 21

[0227] 5-(5-{2-[(4-chlorophenyl)oxy]propyl-2-yl}-1,2,4-oxadiazacyclopentan-3-yl)-1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridine (compound 21)

[0228]

[0229] Following the synthetic method of compound 1, using [(Z)-amino(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)methylpropionic acid as starting materials, a white solid, namely compound 21, was obtained with a yield of 40.1%.

[0230] 1H NMR (500MHz, DMSO-d6): δ8.68(s,1H),7.41(d,J=5.7Hz,2H),7.27(m,3H),7.13(d,J=7.5Hz, 1H), 6.79 (d, J = 8.9Hz, 2H), 5.58 (s, 2H), 3.93 (s, 3H), 2.49 (s, 3H), 2.32 (s, 3H), 1.84 (s, 6H).

[0231] 13 C NMR (126MHz, DMSO-d6): δ179.80,166.25,160.36,153.52,149.69,142.09,137.81,137.05,134.96,129.82, 128.73,128.65,128.53,128.17,125.04,123.28,109.99,104.48,76.62,68.38,33.61,26.36,21.49,12.55.

[0232] HRMS(ESI)m / z calcd for[C 27 H 26 ClN5O3+H] + :504.1797; found:504.1800[M+H] + .

[0233] Example 22

[0234] 1,3-Dimethyl-6-{[(3-methylphenyl)methyl]oxy}-5-{5-[1-(2-methylpropyl-2-yl)pyrazol-4-yl]-1,2,4-oxadiazacyclopentan-3-yl}pyrazolo[3,4-b]pyridine (Compound 22)

[0235]

[0236] Following the synthetic method of compound 1, using [(Z)-amino(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)methylethylenedimethyl]hydroxylamine and 1-(2-methylpropyl-2-yl)pyrazol-4-carboxylic acid as starting materials, a white solid, namely compound 22, was obtained with a yield of 46.5%.

[0237] 1H NMR (500MHz, DMSO-d6): δ8.81-8.72(m,2H),8.22(s,1H),7.45(m,2H),7.30-7.26( m,1H),7.13(d,J=7.6Hz,1H),5.60(s,2H),3.95(s,3H),2.32(s,3H),1.61(s,9H).

[0238] 13 C NMR (126MHz, DMSO-d6): δ170.38,166.24,160.41,149.66,141.98,138.76,137.77,137.18,134.78,129. 63,128.63,128.61,128.38,124.86,110.05,106.45,105.03,68.28,60.15,33.62,29.68,21.54,12.57.

[0239] HRMS(ESI)m / z calcd for[C 25 H 27 N7O2+H] + :458.2299; found:458.2307[M+H] + .

[0240] Example 23

[0241] Experiments on the inhibitory effects of the compound on the proliferation of human pulmonary artery smooth muscle cells, human pulmonary artery endothelial cells and human lung fibroblasts.

[0242] Using KNK437, an Hsp110 expression inhibitor, as a positive control and compound I-7 (patent application number 201910492351.1) as a reference, the CCK-8 assay was used to determine the proliferation inhibition rates of the compounds of this invention on human pulmonary artery smooth muscle cells (HPASMCs), human pulmonary artery endothelial cells (HPAECs), and human lung fibroblasts (HLF1), and the IC50 was calculated for each. 50 value.

[0243] (1) Experimental materials:

[0244] a. Cell lines: Primary human pulmonary artery smooth muscle cells (cultured to passages 3-8), human pulmonary artery endothelial cells, and human lung fibroblasts (which can be used directly for experimental research) were all purchased from ATCC and cultured in DMEM medium containing 10% fetal bovine serum.

[0245] b. Drugs and reagents: 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfonic acid benzene)-2H-tetrazole monosodium salt (CCK-8) and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich; 96-well cell culture plates were purchased from Costar; trypsin, newborn calf serum, fetal calf serum, RPMI-1640, DMEM, and penicillin-streptomycin were purchased from Gibco; EnoGeneCell™ Counting Kit-8 (CCK-8) cell viability assay kit (E1CK-000208-10) was purchased from Nanjing Enjing Biotechnology Co., Ltd.; all other reagents, unless otherwise specified, were purchased from Sigma-Aldrich.

[0246] c. Instruments and consumables: ChemBase CBS-CJ-1FD clean bench; MCO-15AC carbon dioxide incubator (Thermo, USA); C5 multi-functional imager (Bio Tek, USA); inverted fluorescence microscope (Zeiss, Germany), etc.

[0247] (2) Experimental steps:

[0248] a. When the cells are in good condition and the density reaches approximately 90%, seed the cells in 96-well plates. Digest and count the cells to a density of 1 × 10⁶ cells / well. 5 Cell suspension of cells / mL, then at 1×10⁶ cells / well. 4 Cells were seeded with 100 μL of suspension in a 96-well plate, with 3 replicates for each compound.

[0249] b. The 96-well plate was placed in a 37°C, 5% CO2 incubator for 24 hours. Then, the culture medium was aspirated and 100 μL of culture medium containing the corresponding drug concentration was added. At the same time, a blank group and a positive drug group were set up, with 3 replicates for each group.

[0250] c. The 96-well plate was placed in a 37°C, 5% CO2 incubator for 48 hours. 10 μL of CCK-8 solution was added to each well, and the plate was incubated in the incubator for 4 hours.

[0251] d. Absorb the culture medium and measure the OD value at 450 nm using an ELISA reader. Calculate the inhibitory rate and IC50 of each compound against human pulmonary artery smooth muscle cells, human pulmonary artery endothelial cells, and human lung fibroblasts. 50 value.

[0252] e: Dose settings: 100μM; 50μM; 40μM; 30μM; 20μM; 10μM; 5μM; 2.5μM; 0μM.

[0253] (3) Data processing:

[0254] Data were processed using Graphpad Prism 8.0, with the x-axis representing concentration and the y-axis representing inhibition rate. A fitted curve was constructed and the IC50 was calculated. 50 Value, cell proliferation inhibition rate formula:

[0255]

[0256] Table 1. Inhibitory activity of compounds against HPASMCs, HPAECs and HLF1 cell proliferation.

[0257]

[0258]

[0259] The structures of some compounds are as follows:

[0260]

[0261] Experimental results show that the compounds of this invention exhibit more significant anti-proliferative effects against HPASMCs, HPAECs, and HLF1 than the HSP110 expression inhibitor KNK437. This indicates that the compounds protected by this invention possess significant anti-cell proliferation activity under hypoxia and can be applied to the treatment of pulmonary hypertension.

[0262] Example 24

[0263] Experiment on the downstream effects of compounds on heat shock protein 110

[0264] The effects of compound 8 in this invention on the levels of p-STAT3 and c-Myc, downstream proteins of heat shock protein 110, in human pulmonary artery endothelial cells were selected.

[0265] (1) Experimental materials:

[0266] a. Drugs and reagents: 5× lysis buffer: prepared from bromophenol blue, DTT, glycerol, SDS, and Tris-HCl; PBS, TEMED, skim milk, Hsp110 (Abcam, USA), STAT3 (CST, USA), p-STAT3 (CST, USA), c-Myc (CST, USA), β-actin (Proteintech, China); rabbit secondary antibody (Nanjing Enjing Biotechnology Co., Ltd.).

[0267] (2) Instruments and Consumables:

[0268] 6-well plates, PVDF membranes, 5% CO2 constant temperature incubator (Thermo, USA), constant voltage and constant current electrophoresis apparatus (Bio-Rad), electrophoresis tank (Bio-Rad), chemiluminescence imaging system (Bio-Rad), and variable speed horizontal shaker (Kylin-Bell).

[0269] (3) Experimental steps:

[0270] a. Cell protein extraction: HPAECs cells were seeded into 6-well plates, approximately 20 × 10⁴ cells per well. After 12 hours, the cells were fully adhered. Different concentrations of the drug were added to treat the cells, and a control group was set up. After 24 hours, proteins were extracted. The 5× lysis buffer was diluted with PBS to prepare 2× lysis buffer. The medium was aspirated from the 6-well plates, washed 3 times with PBS, placed on ice, and 100 μL of 2× lysis buffer was added. The lysis buffer was stirred with a 200 μL pipette tip until homogeneous. After the lysis buffer became viscous, it was aspirated and placed in a centrifuge tube. The tube was heated at 100 °C for 15 min for denaturation, and then stored at -80 °C.

[0271] b. Preparation of separating and stacking gels: Different separating gels are prepared according to the molecular weight of the protein. Taking 8% and 10% as examples: 20 mL of 8% separating gel composition: H2O 9.3 mL; 29:1 5.3 mL; Tris-HCl-88 5 mL; 10% SDS 200 μL; 10% AP 200 μL; TEMED 12 μL; 20 mL of 10% separating gel composition: H2O 8 mL; 29:1 6.6 mL; Tris-HCl 8.8 5 mL; 10% SDS 200 μL; 10% AP 200 μL; TEMED 12 μL. 6 mL of 5% stacking gel composition: H2O 4.1 mL; 29:1 1 mL; Tris-HCl 6.8 750 μL; 10% SDS 60 μL; 10% AP 60 μL; TEMED 10 μL.

[0272] c. SDS-PAGE electrophoresis: After the stacking gel solidifies, fix it in the electrophoresis tank with clamps, then fill the tank with electrophoresis buffer and remove the comb; adjust the sample loading volume, vortex to mix, and load the samples sequentially, using Mrakker at both ends; after loading the samples, add more electrophoresis buffer and start electrophoresis. Set the voltage to 80-120V, and stop electrophoresis after the proteins are separated.

[0273] d. Transfer: After electrophoresis, immerse the PVDF membrane in the transfer solution, separate the plates, transfer the gel to the filter paper, then cover it with the membrane, cover it with another layer of filter paper and sponge, clamp it with clips and place it in the tank, fill it with transfer solution, connect the positive and negative electrodes, and transfer the membrane at 260mA for 2 hours at low temperature.

[0274] e. Sealing: After the transfer is complete, immerse the PVDF membrane in 5% skim milk for 2 hours to seal it.

[0275] f. Primary antibody incubation: Wash the PVDF membrane with PBST, cut the target band, and place it in the primary antibody solution of the corresponding molecular weight protein. Incubate at 4°C on a shaker for 12 hours.

[0276] g. Secondary antibody incubation: Wash the PBST with PBST, put all the bands into rabbit secondary antibody dilution solution, and incubate at room temperature with shaking for 2 hours.

[0277] 4) Data Analysis

[0278] The PVDF membrane was washed with PBST, photographed on an imager, and quantitative analysis was performed using Image Lab software to calculate the relative protein expression levels.

[0279] The results are as follows Figure 1 As shown. Figure 1 Experimental results showed that under hypoxia, the levels of Hsp110 and its downstream factors were significantly upregulated in human pulmonary artery endothelial cells. Compound 8 did not affect the level of Hsp110 itself, but it could downregulate the levels of its downstream proteins p-STAT3 and c-Myc.

[0280] Example 25

[0281] Hsp110-STAT3 interaction immunoprecipitation assay

[0282] Experimental principle:

[0283] Co-immunoprecipitation (Co-IP) is an effective method for investigating protein-protein interactions within intact cells by utilizing the specific interaction between antigens and antibodies. The principle is as follows: Lysing cells under non-denaturing conditions preserves most intracellular protein interactions. If protein X interacts with protein Y to form a complex, precipitating X with an antibody against X will also precipitate protein Y bound to X. Adding a specific antibody against X to the cell lysate precipitates protein X, and then using Western blotting to detect the presence of protein Y in the precipitated protein, confirms the intracellular interaction of X and Y and the formation of a complex.

[0284] Compound 8, a representative compound of this invention, was selected, and its inhibition of Hsp110-STAT3 interaction was verified by immunoprecipitation experiments.

[0285] (1) Experimental materials: a. Cell lines: Human colorectal cancer cells (HCT-116) and human pulmonary artery endothelial cells (HPAECs) were purchased from ATCC. b. Drugs and reagents: IP lysis buffer, Protein A agarose beads. c. Instruments and consumables: IP tubes, low-temperature centrifuge.

[0286] (2) Experimental procedures:

[0287] a. Cell seeding: Seed HPAECs or HCT116 cells into 6-well plates, approximately 20 × 10⁴ cells per well. After 12 hours of cell attachment, dilute the test compound to the corresponding concentration with culture medium, remove the previous culture medium, and add culture medium containing the drug. Incubate the 6-well plates in an incubator for 48 hours.

[0288] b. Cell lysis: Absorb the culture medium, wash the cells with pre-cooled PBS, remove the PBS, add 500 μL of lysis buffer, scrape off the cells and collect the suspension, and shake on a shaker at 4°C for 15 min.

[0289] c. Add antibody: Centrifuge at 12,000 rpm for 20 min in a 4°C centrifuge, transfer the supernatant to two centrifuge tubes (IP and Input, respectively), add STAT3 antibody, and incubate at 4°C overnight.

[0290] d. Protein extraction: Add 50 μL of Protein Aagarose Beads to an IP tube and incubate at 4°C for 3 h; centrifuge at 8000 rpm for 1 min in a low-temperature centrifuge, discard the supernatant, wash once with IP lysis buffer, centrifuge again for 1 min, and aerate the supernatant; wash the precipitate twice with PBS, centrifuge at 8000 rpm for 1 min, aspirate the PBS, add 50 μL of buffer, denature at 100°C for 10 min, and store the protein at -80°C.

[0291] e. Western blot experiment and data analysis: The experimental steps are the same as in Example 24.

[0292] The results are as follows Figure 2 As shown in the figure. The experimental results indicate that in the IP group, STAT3 can precipitate Hsp110 protein, proving that it interacts with Hsp110. Treatment with compound 8 significantly reduced the protein level of Hsp110 in the precipitate, demonstrating that compound 8 inhibits the interaction between Hsp110 and STAT3 by binding to Hsp110 protein. KNK437 is mainly an inhibitor of Hsp110 expression, but it cannot inhibit the Hsp110-STAT3 interaction.

[0293] The above embodiments are only used to clearly illustrate the present invention and are not intended to limit the scope of the present invention. After understanding the present invention, those skilled in the art should understand that all equivalent modifications of the present invention fall within the scope of the appended claims.

Claims

1. 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and their pharmaceutically usable salts, characterized in that... Its general formula is shown in formula (II): ； R1 is selected from C1-C8 straight-chain or branched alkyl groups, substituted C1-C8 straight-chain or branched alkyl groups, aromatic rings, and substituted aromatic rings; R2 is selected from C1-C8 straight-chain or branched alkyl groups, or substituted C1-C8 straight-chain or branched alkyl groups; R3 is selected from aromatic rings, heterocyclic aromatic rings, substituted aromatic rings, or substituted heterocyclic aromatic rings; The substituents on the substituted C1-C8 straight-chain or branched alkyl group are selected from: halogens and hydroxyl groups; The substituted aromatic heterocycle or the substituents on the substituted aromatic ring are independently selected from: halogens, amino groups, haloalkyl groups, alkyl groups, and hydroxyl groups; The aromatic ring is benzene; the aromatic heterocycle is pyrazole.

2. The 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and their pharmaceutically usable salts according to claim 1, characterized in that, R1 or R2 is independently selected from C1-C4 straight-chain or branched alkyl groups or substituted C1-C4 straight-chain or branched alkyl groups; the substituent is a halogen.

3. The 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and their pharmaceutically usable salts according to claim 1, characterized in that, R1 or R2 is trifluoromethyl independently.

4. The 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and their pharmaceutically usable salts according to claim 1, characterized in that, When the number of substituents on the substituted aromatic heterocycle is 1, the substituents are selected from: -C(CH3)3, -C((CH3)2)CH2CH3, -CH2CH2CH3, -CH2CH3, and -CH3.

5. A 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivative and its pharmaceutically usable salt, characterized in that, The 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives have the following structures: 3-[3-(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 2); 4-[3-(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 3); 3-[3-(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]aniline (compound 4); 2-Chloro-6-[3-(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]aniline (compound 5); 3-[3-(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 8); 4-[3-(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 9); 2-Chloro-6-[3-(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]aniline (Compound 10); 5-(5-{2-[(4-chlorophenyl)oxy]propyl-2-yl}-1,2,4-oxadiazacyclopentan-3-yl)-1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridine (compound 12); 3-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 15); 4-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]phenol (compound 16); 3-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]aniline (compound 17); 2-Chloro-6-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]aniline (compound 19); 6-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazacyclopentan-5-yl]-2-fluoroaniline (compound 20); 5-(5-{2-[(4-chlorophenyl)oxy]propyl-2-yl}-1,2,4-oxadiazacyclopentan-3-yl)-1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridine (compound 21); 1,3-Dimethyl-6-{[(3-methylphenyl)methyl]oxy}-5-{5-[1-(2-methylpropyl-2-yl)pyrazol-4-yl]-1,2,4-oxadiazacyclopentan-3-yl}pyrazolo[3,4-b]pyridine (compound 22).

6. The method for preparing 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives according to any one of claims 1-5, characterized in that, Includes the following steps: S1. The cyanoamino group on compound I is converted to an amidine group to obtain compound II; S2 and compound II react with formic acid substituted with R3 to prepare the 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivative; The structure of compound I is: ; The structure of compound II is as follows: .

7. The use of the 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and pharmaceutically usable salts thereof according to any one of claims 1-5 in the preparation of Hsp110-STAT3 interaction inhibitors.

8. The use of the 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and pharmaceutically usable salts thereof according to any one of claims 1-5 in the preparation of anti-pulmonary hypertension drugs.

9. The use of the 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and pharmaceutically usable salts thereof according to any one of claims 1-5 in the preparation of drugs that downregulate the expression levels of p-STAT3 and c-Myc.

Images