5-[1, 2, 4-oxadiazole]-6-oxygen-pyrazolopyridine derivative as well as preparation method and application thereof

Abstract

The invention relates to a 5-[1, 2, 4-oxadiazole]-6-oxygen-pyrazolopyridine derivative as well as a preparation method and application thereof, the 5-[1, 2, 4-oxadiazole]-6-oxygen-pyrazolopyridine derivative and pharmaceutically available salt thereof have a general formula as shown in a formula (II): # imgabs0 #, and the definition of substituent groups is shown in the specification. The 5-[1, 2, 4-oxadiazole]-6-oxygen-pyrazolopyridine derivative and the pharmaceutically available salt thereof have the effects of inhibiting the interaction of Hsp110-STAT3, so that the levels of p-STAT3 and c-Myc are reduced, and the proliferation, migration and apoptosis resistance of pulmonary artery endothelial cells are inhibited; the pulmonary arterial pressure is reduced, and the pulmonary vascular reconstruction degree is improved, so that the effect of resisting pulmonary arterial hypertension is achieved.

Description

Technical Field

[0001] The present invention relates to 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and pharmaceutically usable salts thereof, and also relates to their preparation methods and applications as medicines, in particular to their applications as medicines for resisting pulmonary hypertension. Background Art

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

[0003] Studies generally believe that the increase in PAP and PVR is 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). Represented by pulmonary artery endothelial cells (HPAECs), pulmonary artery smooth muscle cells (HPASMCs), and pulmonary artery fibroblasts (HLF-1), abnormal proliferation and migration are observed. At the same time, endothelial / mesenchymal transformation, fibrosis, and inflammation promote pulmonary vascular remodeling, leading to thickening of the pulmonary vascular wall, reduction of the inner diameter, increased pulmonary vascular pressure, hypertrophy of the right ventricle, and ultimately death from right heart failure. Due to the similarity between the pathogenesis of pulmonary hypertension and cancer, it is called "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 vital role in mediating changes in pulmonary vascular remodeling.

[0004] At present, the treatment strategies for pulmonary hypertension are nitric oxide (NO), endothelin (ET) and prostacyclin pathways, such as endothelin receptor antagonist bosentan, phosphodiesterase inhibitor sildenafil, prostacyclin analogs, etc. These treatments all focus on vasodilation, which can relieve patient symptoms in a short period of time and improve the patient's short-term survival rate, but do not touch the essence of the pathology, namely pulmonary vascular remodeling, so drug treatment has only a five-year survival rate. Therefore, potential therapeutic targets and drugs are urgently needed to improve vascular remodeling, so as to fundamentally treat pulmonary hypertension.

[0005] Heat shock protein 110 (Hsp110, also known as Hsp105, HSPH1) is an emergency-induced chaperone protein that helps cells survive under stress conditions by preventing the aggregation of misfolded or unfolded proteins. A large number of studies have shown that Hsp110 is upregulated in a variety of 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, especially the signal transducer and activator of transcription 3 (STAT3) pathway, thereby regulating tumor growth.

[0006] The prior art with application number 201910492351.1 discloses a compound of the 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; R4 is selected from hydrogen, the following structural fragments and their pharmaceutically acceptable salts: -NO-NH2 Wherein, R5 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; Z is selected from CH2, O, N.

[0007]

[0008] The patent discloses the use of the compound of formula (I) for preparing drugs against pulmonary fibrosis, but no one has used it to treat pulmonary hypertension.

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

[0010]

[0011] This type of compound is a stimulator of soluble guanylate cyclase, wherein R1 represents thiophene or substituted pyridyl, R2 represents -NH2, -NHCH3, -NHC(=O)CH(CH3)2, R3 is hydrogen or C1-C4 alkyl, and R4 is C1-C6 alkyl. The invention also discloses the preparation of the compound and its application as a drug, especially as a drug for treating cardiovascular diseases, such as anti-pulmonary hypertension. However, its effect still needs to be improved. Summary of the invention

[0012] The purpose of the present invention is to provide a 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivative and a preparation method thereof and an application thereof in preparing a drug for treating pulmonary arterial hypertension.

[0013] In order to achieve the above object, the technical solution of the present invention is as follows:

[0014] 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and pharmaceutically acceptable salts thereof, the general formula of which is shown in formula (II):

[0015]

[0016] wherein R1 or R2 is independently selected from a C1-C8 straight or branched alkyl group, or a substituted C1-C8 straight or branched alkyl group, an aromatic ring, an aromatic heterocycle, a substituted aromatic ring, or a substituted aromatic heterocycle;

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

[0018] Wherein, the substituent on the substituted C1-C8 straight or branched alkyl group is selected from the group consisting of halogen, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, and hydroxyl;

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

[0020] In one preferred embodiment, R1 or R2 is independently selected from C1-C4 straight chain or branched alkyl or substituted C1-C4 straight chain or branched alkyl; and the substituent is 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 a preferred embodiment, the aromatic ring is selected from benzene, naphthalene, anthracene and phenanthrene.

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

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

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

[0026] The number of substituents on the substituted aromatic heterocycle is arbitrarily 1-5, the positions of the substituents are arbitrarily 1-5, and the substituents are selected from: halogen, amino, haloalkyl, alkyl, hydroxyl, 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 substituent is selected from: halogen, amino, halogenated C1-C4 alkyl, C1-C4 alkyl, hydroxyl, C1-C4 hydroxyalkyl.

[0028] In one preferred embodiment, R3 is selected from a substituted aromatic ring; when the number of substituents is 2, and the position of the substituents is any one of positions 2-6, the substituents are selected from: halogen, amino, halogenated C1-C4 alkyl, C1-C4 alkyl, hydroxyl, C1-C4 hydroxyalkyl.

[0029] In one preferred embodiment, R3 is selected from a substituted aromatic ring; when the number of substituents is 2, one of the substituents is at position 1, and the other substituent is arbitrarily 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 alkyleneoxy, 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)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 substituent is 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 acceptable 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 pharmaceutical 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, mucate, malonate, benzoate, DL-malate, (±) lactate, L-(+)-lactate, D-(+)-lactate, pamoate, D-α-galacturonate, glycerate, DL-cysteine, D-cysteine, L-cysteine, (4S)-hydroxy-L-proline, Cyclopropane-1,1-dicarboxylate, 2,2-methylmalonate, tyrosinate, proline, fumarate, 1-hydroxy-2-naphthoate, 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, isethionate, 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, tribasic calcium phosphate, hexafluorophosphate, vinyl phosphate, 2-hydroxyethyl phosphate and phenyl phosphate.

[0041] In one preferred embodiment, the structure of 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-oxadiazol-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-oxadiazol-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-oxadiazol-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-oxadiazol-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-oxadiazol-5-yl]aniline (Compound 5);

[0047] 5-(5-{2-[(4-chlorophenyl)oxy]propan-2-yl}-1,2,4-oxadiazol-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-oxadiazol-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-oxadiazol-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-oxadiazol-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-oxadiazol-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-oxadiazol-5-yl]aniline (Compound 11);

[0053] 5-(5-{2-[(4-chlorophenyl)oxy]propan-2-yl}-1,2,4-oxadiazol-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-oxadiazacyclopentyl-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-oxadiazol-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-oxadiazol-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-oxadiazol-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-oxadiazol-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-oxadiazol-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-oxadiazol-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-oxadiazol-5-yl]-2-fluoroaniline (Compound 20);

[0062] 5-(5-{2-[(4-chlorophenyl)oxy]propan-2-yl}-1,2,4-oxadiazacyclopentyl-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-methylpropan-2-yl)pyrazol-4-yl]-1,2,4-oxadiazacyclopentyl-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 derivatives, comprising the following steps:

[0065] S1, aminolyzing the cyano group on compound I into amidine group to obtain compound II;

[0066] S2, compound II reacts with formic acid substituted by R3 to obtain the 5-[1,2,4-oxadiazole]-6-oxy-pyrazolopyridine derivative;

[0067] The structure of compound I is:

[0068] The structure of compound II is:

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

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

[0071]

[0072] wherein R1, R2, and R3 are as defined 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 pharmaceutically acceptable salts thereof 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 pharmaceutically acceptable salts thereof 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 pharmaceutically acceptable salts thereof in the preparation of drugs for downregulating 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 pharmaceutically acceptable salts thereof in the preparation of drugs for inhibiting the proliferation, migration and / or apoptosis resistance of pulmonary artery endothelial cells.

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

[0078] The 5-[1,2,4-oxadiazole]-6-oxy-pyrazolopyridine derivatives and pharmaceutically acceptable salts thereof of the present invention have the effects of inhibiting 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 effect of resisting pulmonary hypertension.

[0079] Compared with the prior art, the present invention has the following beneficial effects:

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

[0081] 2. The compounds synthesized in the present invention have an anti-pulmonary hypertension effect and can be used as clinical drugs for the treatment of related diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0082] Figure 1 is the expression level of Hsp110 in human pulmonary artery endothelial cells after 12 hours of hypoxia under normoxia, hypoxia, or compound treatment in Example 24;

[0083] Figure 2 is the p-STAT3 expression level in human pulmonary artery endothelial cells after 12 hours of hypoxia under normoxia, hypoxia, or compound treatment in Example 24;

[0084] Figure 3 is the c-Myc expression level in human pulmonary artery endothelial cells after 12 hours of hypoxia under normoxia, hypoxia, or compound treatment in Example 24;

[0085] Figure 4 This is the binding of Hsp110 and STAT3 in human pulmonary artery endothelial cells under normoxia and after treatment with different concentrations of compounds for 12 hours in Example 25, with the ratio of the Hsp110 level to the STAT3 level in the IP group as a reference. DETAILED DESCRIPTION

[0086] The present invention is 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-oxadiazol-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 in a 30 mL microwave tube, and then 15 mL DMF was added to dissolve it, 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 [(Z)-amino(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)methylidene]hydroxylamine (0.774 mmol) was added, and then the reaction solution was transferred to a microwave and reacted at 120°C for 40 min. TLC monitored that the reaction of the raw materials was complete and the reaction solution was dark yellow. The reaction solution was cooled to room temperature, poured into 50 mL of water, and stirred at room temperature for 30 min. A white suspension was present in the beaker. The mixture was extracted with ethyl acetate three times, 50 mL each time. The organic phases were combined and washed with saturated brine twice, 100 mL each time. The organic phase was then dried over 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 a white solid, which was recrystallized with anhydrous ethanol. First, 20 mL of ethanol was added to a 100 mL eggplant-shaped bottle. It was found that the sample was basically insoluble at room temperature and was a white suspension. After reflux at 80°C for 15 min, the solid was still not completely dissolved. Then 10 mL of ethanol was added to the bottle and refluxed for 15 min. The solution in the bottle gradually became clear, but a small amount of solid was still not dissolved. After adding 5 mL of ethanol, reflux was continued for 10 min. It was found that the sample in the bottle was completely dissolved. The reaction bottle was left to stand at room temperature overnight. A white solid precipitated in the bottle. The white filter cake was filtered and dried to obtain 0.19 g of a white solid, namely 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 calculation 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-oxadiazol-5-yl]phenol (Compound 2)

[0096]

[0097] According to the synthesis method of compound 1, compound [(Z)-amino(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)methylidene]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 calculation 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-oxadiazol-5-yl]phenol (Compound 3)

[0103]

[0104] According to the synthesis method of compound 1, compound [(Z)-amino(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)methylidene]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 calculation 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-oxadiazol-5-yl]aniline (Compound 4)

[0110]

[0111] According to the synthesis method of compound 1, compound [(Z)-amino(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)methylidene]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 calculation 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-oxadiazol-5-yl]aniline (Compound 5)

[0117]

[0118] According to the synthesis method of compound 1, compound [(Z)-amino(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)methylidene]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 calculation for[C 29 H 23 ClN6O2+H] + :523.1587; found:523.1592[M+H] + .

[0122] Example 6

[0123] 5-(5-{2-[(4-chlorophenyl)oxy]propan-2-yl}-1,2,4-oxadiazol-3-yl)-3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridine (Compound 6)

[0124]

[0125] According to the synthesis method of compound 1, using compound [(Z)-amino(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)methylidene]hydroxylamine and 2-[(4-chlorophenyl)oxy]-2-methylpropionic acid as raw materials, a white solid, namely compound 6, was obtained 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 calculation 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-oxadiazol-5-yl]phenol (Compound 7)

[0131]

[0132] According to the synthesis method of compound 1, compound [(Z)-amino(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)methylidene]hydroxylamine and 2-hydroxybenzoic acid were used as raw materials to obtain a white solid, namely compound 7, 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 calculation 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-oxadiazol-5-yl]phenol (Compound 8)

[0138]

[0139] According to the synthesis method of compound 1, compound [(Z)-amino(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)methylidene]hydroxylamine and 3-hydroxybenzoic acid were used as raw materials to obtain a white solid, namely compound 8, 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 calculation 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-oxadiazol-5-yl]phenol (Compound 9)

[0145]

[0146] According to the synthesis method of compound 1, compound [(Z)-amino(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)methylidene]hydroxylamine and p-hydroxybenzoic acid were used as raw materials to obtain a white solid, namely compound 9, 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 calculation 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-oxadiazol-5-yl]aniline (Compound 10)

[0152]

[0153] According to the synthesis method of compound 1, using compound [(Z)-amino(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)methylidene]hydroxylamine and 2-amino-3-chlorobenzoic acid as raw materials, a khaki solid, namely compound 10, was obtained 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 calculation for[C 24 H 18 ClF3N6O2+H] + :515.1205; found:515.1208[M+H] + .

[0157] Embodiment 11

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

[0159]

[0160] According to the synthesis method of compound 1, using compound [(Z)-amino(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)methylidene]hydroxylamine and 3-aminobenzoic acid as raw materials, a khaki 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 calculation for[C 24 H 19 F3N6O2+H] + :481.1594; found:481.1601[M+H] + .

[0164] Example 12

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

[0166]

[0167] According to the synthesis method of compound 1, using compound [(Z)-amino(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)methylidene]hydroxylamine and 2-[(4-chlorophenyl)oxy]-2-methylpropionic acid as raw 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 calculation 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-oxadiazol-3-yl)-3-(trifluoromethyl)pyrazolo[3,4-b]pyridine

[0173] According to the synthesis method of compound 1, compound [(Z)-amino(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)methylidene]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 calculation for[C 18 H 14 F3N5O2+H] + :390.1100; found:390.1108[M+H] + .

[0177] Embodiment 14

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

[0179]

[0180] According to the synthesis method of compound 1, using [(Z)-amino(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)methylidene]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 calculation for[C 24 H 21 N5O3+H] +:428.1717; found:428.1722[M+H] + .

[0184] Embodiment 15

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

[0186]

[0187] According to the synthesis method of compound 1, using [(Z)-amino(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)methylidene]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 calculation 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-oxadiazol-5-yl]phenol (Compound 16)

[0193]

[0194] According to the synthesis method of compound 1, using [(Z)-amino(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)methylidene]hydroxylamine and 4-hydroxybenzoic acid as raw 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 calculation for[C 24 H 21 N5O3+H] + :428.1717; found:428.1721[M+H] + .

[0198] Embodiment 17

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

[0200]

[0201] According to the synthesis method of compound 1, [(Z)-amino(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)methylidene]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 calculation for[C 24 H 22 N6O2+H] + :427.1877; found:427.1882[M+H] + .

[0205] Embodiment 18

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

[0207]

[0208] According to the synthesis method of compound 1, using [(Z)-amino(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)methylidene]hydroxylamine and 4-aminobenzoic acid as raw materials, a light 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 calculation for[C 24 H 22 N6O2+H] + :427.1877; found:427.1883[M+H] + .

[0212] Embodiment 19

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

[0214]

[0215] According to the synthesis method of compound 1, [(Z)-amino(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)methylidene]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 calculation for[C 24 H 21 ClN6O2+H] + :461.1487; found:461.1490[M+H] + .

[0219] Embodiment 20

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

[0221]

[0222] According to the synthesis method of compound 1, using (Z)-amino(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)methylidene]hydroxylamine and 2-amino-3-fluoro-benzoic acid as raw materials, a light 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 calculation for[C 24 H 21 FN6O2+H] + :445.1783; found:445.1784[M+H] + .

[0226] Embodiment 21

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

[0228]

[0229] According to the synthesis method of compound 1, using [(Z)-amino(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)methylidene]hydroxylamine and 2-[(4-chlorophenyl)oxy]-2-methylpropionic acid as raw 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 calculation for[C 27 H 26 ClN5O3+H] + :504.1797; found:504.1800[M+H] + .

[0233] Embodiment 22

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

[0235]

[0236] According to the synthesis method of compound 1, using [(Z)-amino(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)methylidene]hydroxylamine and 1-(2-methylprop-2-yl)pyrazole-4-carboxylic acid as raw 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 calculation for[C 25 H 27 N7O2+H] + :458.2299; found:458.2307[M+H] + .

[0240] Embodiment 23

[0241] The compound was tested for its inhibition of proliferation of human pulmonary artery smooth muscle cells, human pulmonary artery endothelial cells and human lung fibroblasts.

[0242] The expression inhibitor of Hsp110, KNK437, was used as a positive drug and compound I-7 (patent application number 201910492351.1) was used as a reference. The CCK-8 method was used to determine the proliferation inhibition rate of the compound of the present invention on human pulmonary artery smooth muscle cells (HPASMCs), human pulmonary artery endothelial cells (HPAECs) and human lung fibroblasts (HLF1), and the IC 50 value.

[0243] (1) Experimental materials:

[0244] a. Cell lines: Primary human pulmonary artery smooth muscle cells (cultured to passage 3-8), human pulmonary artery endothelial cells and human lung fibroblasts (can be used directly for experimental research) were 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; 96-well cell culture plates were purchased from Costar; trypsin, newborn calf serum, fetal bovine serum, RPMI-1640, DMEM and penicillin-streptomycin were purchased from Gibco; EnoGeneCellTM Counting Kit-8 (CCK-8) cell viability detection kit (E1CK-000208-10) was purchased from Nanjing Enjing Biotechnology Co., Ltd.; other reagents were purchased from Sigma except for those specified.

[0246] c. Instruments and consumables: ChemBase CBS-CJ-1FD clean bench; MCO-15AC carbon dioxide incubator (Thermo, USA); C5 multifunctional 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 about 90%, plate the cells in a 96-well plate. Digest and count the cells to 1×10 5 Then, 1×10 cells / mL of cell suspension was added to each well. 4 100 μL of the suspension was inoculated into a 96-well plate, and 3 replicate wells were set for each compound.

[0249] b. Place the 96-well plate in a 37°C, 5% CO2 incubator for 24 hours, then aspirate the culture medium and add 100 μL of culture medium containing the corresponding drug concentration. Set up a blank group and a positive drug group at the same time, with 3 replicates in each group.

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

[0251] d. Aspirate the culture medium and measure the OD value at 450nm using an enzyme marker to calculate the inhibition rate, IC value and inhibitory rate of each compound on human pulmonary artery smooth muscle cells, human pulmonary artery endothelial cells and human pulmonary fibroblasts. 50 value.

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

[0253] (3) Data processing:

[0254] The data were processed using Graphpad Prism 8.0, with the horizontal axis representing the concentration and the vertical axis representing the inhibition rate. The curve was fitted and the IC 50 Value, cell proliferation inhibition rate formula:

[0255]

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

[0257]

[0258]

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

[0260]

[0261] The experimental results show that the anti-proliferation effect of the compound of the present invention on HPASMCs, HPAECs and HLF1 is more obvious than that of the HSP110 expression inhibitor KNK437, indicating that the compound protected by the present invention has a significant anti-cell proliferation effect under hypoxia and can be used in the treatment of pulmonary hypertension.

[0262] Embodiment 24

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

[0264] The effect of compound 8 in the present invention on the levels of p-STAT3 and c-Myc, downstream proteins of heat shock protein 110, in human pulmonary artery endothelial cells was 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), and β-actin (Proteintech, China); rabbit secondary antibody (Nanjing Enjing Biotechnology Co., Ltd.).

[0267] (2) Instruments and consumables:

[0268] 6-well plate, PVDF membrane, 5% carbon dioxide constant temperature incubator (Thermo, USA), constant voltage and constant current electrophoresis apparatus (Bio-Rad), electrophoresis tank (Bio-Rad), chemiluminescence imager (Bio-Rad), variable speed horizontal shaker (Kylin-Bell).

[0269] (3) Experimental steps:

[0270] a. Cell protein extraction: HPAECs cells were plated into 6-well plates, with about 20×104 cells per well. After 12 hours, the cells were completely attached to the wall, and different concentrations of drugs were added to treat the cells, and a blank group was set up. After 24 hours, protein was extracted. The 5× lysis buffer was diluted with PBS to 2× lysis buffer, and the medium in the 6-well plate was aspirated, washed 3 times with PBS, placed on ice, and 100μL of 2× lysis buffer was added. The lysis buffer was stirred with a 200μL pipette tip to make it uniform; when the lysis buffer became viscous, it was aspirated and placed in a centrifuge tube, heated at 100℃ for 15min to denature, and then stored in a -80℃ refrigerator.

[0271] b. Prepare separation gel and stacking gel: Prepare different separation gels according to the molecular weight of the protein. Take 8% and 10% as examples: 20mL of 8% separation gel composition: H2O 9.3mL; 29:1 5.3mL; Tris-HCl-88 5mL; 10% SDS200μL; 10% AP 200μL; TEMED 12μL; 20mL of 10% separation gel composition: H2O 8mL; 29:1 6.6mL; Tris-HCl8.8 5mL; 10% SDS200μL; 10% AP 200μL TEMED 12μL. 6mL of 5% stacking gel composition: H2O 4.1mL; 29:1 1mL; Tris-HCl 6.8 750μL; 10% SDS 60μL; 10% AP 60μL; TEMED 10μL.

[0272] c. SDS-PAGE electrophoresis: After the concentrated gel solidifies, fix it with a clamp and place it in the electrophoresis tank. Then fill the tank with electrophoresis solution and pull out the comb. Adjust the sample loading volume, vortex and mix well, then load the samples in sequence, with Mraker at both ends. After adding the samples, add electrophoresis solution and start electrophoresis. Set the voltage to 80-120V, and stop electrophoresis after the protein is separated.

[0273] d. Transfer: After the electrophoresis is finished, soak the PVDF membrane in the transfer solution, separate the plates, transfer the gel to the filter paper, and then cover the membrane with a layer of filter paper and sponge. Clamp it with a clip and place it in the tank. After filling it with transfer solution, connect the positive and negative electrodes, and transfer the membrane at 260mA at low temperature for 2 hours.

[0274] e. Blocking: After transfer, soak the PVDF membrane in 5% skim milk for 2 hours.

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

[0276] g. Secondary antibody incubation: Wash the PBST with PBST, place all bands in the rabbit secondary antibody dilution solution, and incubate with shaking at room temperature 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 expression of the protein.

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

[0280] Embodiment 25

[0281] Hsp110-STAT3 interaction co-immunoprecipitation assay

[0282] Experimental principle:

[0283] Co-immunoprecipitation (Co-IP) experiments use antigen-antibody specificity to study protein-protein interactions, and are an effective means of exploring whether proteins interact in intact cells. The principle is: Lysing cells under non-denaturing conditions can retain most of the protein-protein interactions in the cell; if X and Y proteins interact to form a complex, X is precipitated with X's antibody, and protein Y bound to X will also precipitate. Add X's specific antibody to the cell lysate to precipitate protein X, and then use Western blotting to detect whether Y protein exists in the precipitated protein. If it exists, it proves that XY interacts and forms a complex in the cell.

[0284] Representative compound 8 of the present invention was selected to verify whether it inhibited the Hsp110-STAT3 interaction through immunoprecipitation experiment

[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 in 6-well plates, with about 20×104 cells per well. After 12 hours of cell attachment, dilute the compound to be tested with culture medium to the corresponding concentration, aspirate the previous culture medium, and add the medium containing the drug. Place the 6-well plate in an incubator and incubate for 48 hours.

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

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

[0290] d. Protein extraction: Add 50 μL Protein Aagarose Beads to the 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 remove the supernatant; wash the precipitate twice with PBS, centrifuge at 8000 rpm for 1 min, absorb the PBS, add 50 μL buffer, denature at 100°C for 10 min, and store the protein in a -80°C refrigerator.

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

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

[0293] The contents described in 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 will realize that various equivalent modifications of the present invention fall within the scope of the claims attached to the present invention.

Claims

1. 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and pharmaceutically acceptable salts thereof, characterized in that: Its general formula is shown in formula (II): 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 heterocycle, substituted aromatic ring or substituted aromatic heterocycle; R3 is selected from hydrogen, aromatic ring, aromatic heterocycle, substituted aromatic ring or substituted aromatic heterocycle; Wherein, the substituent on the substituted C1-C8 straight chain or branched alkyl is selected from: halogen, amino, carboxyl, phenyl, benzyl, Phenyloxy, =O, hydroxyl; The substituents on the substituted heterocyclic ring or substituted aromatic ring are independently selected from the group consisting of halogen, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, and alkoxy.

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

3. The 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and pharmaceutically acceptable salts thereof according to claim 1, characterized in that: The aromatic ring is selected from benzene, naphthalene, anthracene, and phenanthrene; the aromatic heterocyclic ring is selected from pyrrole, furan, thiophene, imidazole, thiazole, oxazole, pyrazole, isoxazole, thiadiazole, oxadiazole, tetrazole, pyridine, pyrimidine, pyrazine, pyridazine, purine, Quinoline, isoquinoline, indole, acridine, carbazole and bioisosteres of the above ring systems.

4. The 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and pharmaceutically acceptable salts thereof according to claim 1, characterized in that: The number of substituents on the substituted aromatic heterocyclic ring is arbitrarily 1-5, and the positions of the substituents are arbitrarily 1-5, The substituents are selected from: halogen, amino, haloalkyl, alkyl, hydroxyl, hydroxyalkyl, alkoxy; preferably, when the number of substituents is 1, the substituents are selected from: -C(CH3)3, -C((CH3)2)CH2CH3, -CH2CH2CH3, -CH2CH3, -CH3.

5. The 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivative and its pharmaceutically acceptable salt according to any one of claims 1 to 5, characterized in that: R3 is selected from substituted aromatic rings; when the number of substituents is 2, one of the substituents is at position 1, and the other substituent is arbitrarily 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 alkyleneoxy, C1-C5 alkylene; preferably, 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-.

6. The 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and pharmaceutically acceptable salts thereof according to claim 5, characterized in that: The structure of the 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivative has the following structure: 2-[3-(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazol-5-yl]phenol (Compound 1); 3-[3-(3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazol-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-oxadiazol-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-oxadiazol-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-oxadiazol-5-yl]aniline (Compound 5); 5-(5-{2-[(4-chlorophenyl)oxy]propan-2-yl}-1,2,4-oxadiazol-3-yl)-3-methyl-6-{[(3-methylphenyl)methyl]oxy}-1-phenylpyrazolo[3,4-b]pyridine (Compound 6); 2-[3-(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazol-5-yl]phenol (Compound 7); 3-[3-(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazol-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-oxadiazol-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-oxadiazol-5-yl]aniline (Compound 10); 3-[3-(1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazol-5-yl]aniline (Compound 11); 5-(5-{2-[(4-chlorophenyl)oxy]propan-2-yl}-1,2,4-oxadiazol-3-yl)-1-methyl-6-{[(3-methylphenyl)methyl]oxy}-3-(trifluoromethyl)pyrazolo[3,4-b]pyridine (Compound 12); 1-Methyl-6-{[(3-methylphenyl)methyl]oxy}-5-(1,2,4-oxadiazacyclopentyl-3-yl)-3-(trifluoromethyl)pyrazolo[3,4-b]pyridine (Compound 13); 2-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazol-5-yl]phenol (Compound 14); 3-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazol-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-oxadiazol-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-oxadiazol-5-yl]aniline (Compound 17); 4-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazol-5-yl]aniline (Compound 18); 2-Chloro-6-[3-(1,3-dimethyl-6-{[(3-methylphenyl)methyl]oxy}pyrazolo[3,4-b]pyridin-5-yl)-1,2,4-oxadiazol-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-oxadiazol-5-yl]-2-fluoroaniline (Compound 20); 5-(5-{2-[(4-chlorophenyl)oxy]propan-2-yl}-1,2,4-oxadiazacyclopentyl-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-methylpropan-2-yl)pyrazol-4-yl]-1,2,4-oxadiazacyclopentyl-3-yl}pyrazolo[3,4-b]pyridine (Compound 22).

7. The method for preparing 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives according to any one of claims 1 to 6, characterized in that: The following steps are involved: S1, aminolyzing the cyano group on compound I into amidine group to obtain compound II; S2, compound II reacts with formic acid substituted by R3 to obtain the 5-[1,2,4-oxadiazole]-6-oxy-pyrazolopyridine derivative; the structure of compound I is: The structure of compound II is:

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

9. Use of the 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and pharmaceutically acceptable salts thereof according to any one of claims 1 to 6 in the preparation of drugs for treating pulmonary hypertension.

10. Use of the 5-[1,2,4-oxadiazole]-6-oxo-pyrazolopyridine derivatives and pharmaceutically acceptable salts thereof according to any one of claims 1 to 6 in the preparation of drugs for downregulating the expression levels of p-STAT3 and c-Myc.

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