Preparation of 3-amino indazole covalent compound and application of 3-amino indazole covalent compound in treatment of Parkinson's disease

Abstract

The invention relates to preparation and application of a 3-amino indazole covalent compound, and belongs to the technical field of medicines. The technical problem to be solved by the invention is to provide the 3-amino indazole covalent inhibitor compound targeting JNK3. The compound comprises a compound as shown in the specification or a pharmaceutically acceptable salt thereof. The compound or the pharmaceutically acceptable salt thereof disclosed by the invention is a JNK3 selective covalent inhibitor, has the activity of inhibiting JNK3 protein and the activity of resisting Parkinson's disease, and shows an excellent neuroprotective effect. The compound disclosed by the invention is preferably an anti-Parkinson's disease compound, and is characterized by being a JNK3 covalent inhibitor with reasonable blood-brain barrier permeability, low toxicity and high selectivity. # imgabs0 wherein R1 is # imgabs1 # imgabs2 # imgabs2 #, R2 is # imgabs3 # imgabs4 #, and R3 is # imgabs5 # imgabs6 #

Description

Technical Field

[0001] The invention relates to the preparation and application of a 3-aminoindazole covalent compound, and belongs to the technical field of medicine. Background Art

[0002] c-Jun N-terminal kinase 3 (JNK3) is a subtype of c-Jun N-terminal kinases (JNKs) that is mainly expressed in the brain. It plays a vital role in neurodegeneration and is involved in learning, brain development, and axon formation. JNK3 is activated through a cascade phosphorylation process, which in turn regulates transcription factors, participates in cell differentiation and apoptosis, and responds to stress stimuli. JNK3 is one of the main factors leading to neuronal dysfunction. The activation of JNK3 is crucial in inducing apoptosis and neuronal death in neurodegenerative diseases such as Parkinson's disease (PD), Alzheimer's disease (AD), and Huntington's disease (HD).

[0003] Due to the similarity of amino acid sequences of JNK subtypes (JNK1, JNK2 and JNK3), it is difficult to find a selective agent for JNK3 inhibitors, and no such inhibitors have been used for PD treatment. The present invention designs and synthesizes a series of 3-aminoindazole covalent inhibitors targeting JNK3. Such inhibitors can exhibit good neuroprotective activity in nerve cells by covalently inhibiting JNK3 protein, so the development of such covalent inhibitors has a very broad research prospect. Summary of the invention

[0004] The technical problem solved by the present invention is to provide a 3-aminoindazole covalent inhibitor targeting JNK3.

[0005] The present invention provides the following compounds or pharmaceutically acceptable salts thereof:

[0006]

[0007] Among them, R 1 for

[0008] R 2 for

[0009] R 3 for

[0010] The present invention also provides use of the above compound or a pharmaceutically acceptable salt thereof in preparing an anti-Parkinson's disease drug.

[0011] Furthermore, the anti-Parkinson's drug is preferably an inhibitor drug targeting JNK3.

[0012] The compound prepared by the present invention or its pharmaceutically acceptable salt can be used as a covalently targeted JNK3 inhibitor, has neuroprotective activity, and can effectively protect nerve cells. The compound of the present invention has a protective effect on nerve cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 After treatment with compound 13, MPP + The protective activity of PD-induced cell model and MPTP-induced PD mouse model. DETAILED DESCRIPTION

[0014] The present invention provides the following compounds or pharmaceutically acceptable salts thereof:

[0015]

[0016]

[0017] Among them, R 1 for

[0018] R 2 for

[0019] R 3 for

[0020] Below are some preferred structures of the compounds of the present invention.

[0021]

[0022]

[0023] The present invention also provides use of the above compound or a pharmaceutically acceptable salt thereof in preparing an anti-Parkinson's disease drug.

[0024] Furthermore, the anti-Parkinson's drug is preferably an inhibitor drug that covalently targets JNK3.

[0025] The present invention also provides a pharmaceutical composition, which is a preparation comprising an effective dose of the above compound or a pharmaceutically acceptable salt thereof.

[0026] The compounds of the present invention can be prepared in the following forms by methods known in the art: tablets, capsules, aqueous or oily solutions, suspensions, emulsions, creams, ointments, gels, nasal sprays, suppositories, finely dispersed powders or aerosols or sprays for inhalation, sterile aqueous or oily solutions or suspensions or sterile emulsions for parenteral administration (including intravenous, intramuscular or infusion). Sterile water or water-propylene glycol solutions can be used as solvents to prepare liquid preparations, and the active ingredients can also be formulated in polyethylene glycol aqueous solutions. Aqueous solutions for oral administration can be prepared by dissolving the active ingredients in water and adding suitable colorants, flavoring agents, stabilizers and thickeners as needed. Aqueous suspensions for oral use can be dispersed in water with finely dispersed active ingredients together with viscous substances, such as natural synthetic gums, resins, methylcellulose, carboxymethylcellulose and other suspending agents known in the pharmaceutical field.

[0027] The pharmaceutical composition may be in unit dosage form. In these forms, the composition is divided into unit doses containing appropriate quantities of the active ingredient. The unit dosage form may be a packaged preparation containing discrete quantities of the preparation, such as boxed tablets, capsules, and powders in vials or ampoules. The unit dosage form may also be a capsule, cachet, or tablet or it may be an appropriate number of any of these packaged forms.

[0028] The active ingredient of the pharmaceutical composition of the present invention may be the compound of the present invention alone, or it may be combined with other Parkinson's disease treating compounds as active ingredients.

[0029] In the process of treating Parkinson's disease, the pharmaceutical composition of the present invention can be used in combination with other drugs for treating Parkinson's disease, for example, in combination with non-ergot dopamine receptor (DR) agonists, monoamine oxidase B (MAO-B) inhibitors, amantadine, compound levodopa, etc.

[0030] In the treatment of Parkinson's disease, such combination therapy may be achieved by simultaneous, sequential or separate administration of the various therapeutic components. Such combination products utilize the compounds of this invention within their effective dosage ranges and the other pharmaceutically active agent within their approved dosage ranges.

[0031] The specific implementation modes of the present invention are further described below in conjunction with embodiments, but the present invention is not limited to the scope of the embodiments.

[0032] Example 1 Synthesis of Preferred Compounds

[0033] The preferred compound is synthesized using the following reaction formula:

[0034] Synthesis route 1: Synthesis of compound 1-4

[0035]

[0036] (a) Acryloyl chloride, K2CO3, DCM, 0℃-rt, overnight; (b) DIPEA, KI, DMF, 80℃, overnight; (c) DCM, TFA, rt, 2-3h; (d) 3,4-dihydro-2H-pyran (DHP), p-toluenesulfonic acid (p-TSA, cat.), DCM, 0℃-rt, 6h; (e) Piperazine, HATU, DIPEA, DCM, rt, 30min; (f) (1) Intermediate 1a-c, HATU, DIPEA, DMF, rt, 30min; (2) TFA, DCM, rt, 2h. (g) (1) Acryloyl chloride, TEA, DMF, rt, 30min; (2) TFA, DCM, rt, 2h.

[0037] Synthesis route 2: Synthesis of compound 5-13

[0038]

[0039] (a) Acrylamide, K2CO3, DCM, 0℃-rt, overnight; (b) (1) Intermediate 5, HATU, DIPEA, DMF, rt, 30min; (2) TFA, DCM, rt, 2h.

[0040] Synthesis route 3: Synthesis of compounds 14-25

[0041]

[0042]

[0043] (a) Acid chloride derivative, K2CO3, DCM, 0℃-rt, overnight; (b) (1) Intermediate 5, HATU, DIPEA, DMF, rt, 30min; (2) TFA, DCM, rt, 2h; (c) Substituted piperazine, HATU, DIPEA, DCM, rt, 30min; (d) (1) Intermediate 7i, HATU, DIPEA, DMF, rt, 30min; (2) TFA, DCM, rt, 2h.

[0044] Synthesis route 4: Synthesis of compounds 26-30

[0045]

[0046] (a) DIPEA, KI, DMF, 80°C, overnight; (b) DCM, TFA, rt, 2-3h; (c) DHP, cat.p-TSA, DCM, 0°C-rt, 6h; (d) piperazine, HATU, DIPEA, DCM, rt, 30min; (e) (1) intermediate 7i, HATU, DIPEA, DMF, rt, 30min; (2) TFA, DCM, rt, 2h.

[0047] 1. General Synthesis of Compounds 1-4

[0048] Add aminobenzoic acid (1 eq) and potassium carbonate (K2CO3, 2.5 eq) to DMF and stir overnight in an ice bath. Then slowly add acryloyl chloride (1.2 eq) to the reaction mixture and stir overnight. TLC monitors the reaction to be complete and vacuum concentrate to obtain a white solid. Add a small amount of water to the solid to dissolve, extract with EA to remove fat-soluble impurities, collect the water layer, adjust the pH to 3-4, precipitate a white solid, filter with suction, and dry to obtain intermediates 1a-1c.

[0049] 4-Chloro-1H-indazole-3-amine (3.00 g, 17.90 mmol) was dissolved in DMF (30 mL), and DIPEA (9.50 mL, 53.70 mmol), cat.KI (594.30 mg, 0.2 mmol) and tert-butyl p-chlorobenzoate (4.46 g, 19.69 mmol) were added thereto in sequence, stirred, heated, and reacted in an 80°C oil bath overnight. TLC monitored the completion of the reaction, and water (200 mL) and EA (200 mL×2) were added thereto for extraction, and the organic phases were combined. The organic phase was then washed three times with water (300 mL), EA was combined, washed with saturated brine (200 mL), dried over anhydrous Na2SO4, and EA was removed by distillation under reduced pressure to obtain a yellow oily crude product. The crude product was purified by column chromatography, PE:EA=5:1; 4:1, to obtain intermediate 2 (white solid, 4.91 g), with a yield of 76.65%.

[0050] Intermediate 2 (13.72 mmol) was dissolved in DCM (30 mL), and TFA (DCM:TFA=3:1) was added thereto, and the reaction was carried out at room temperature for 2-3 h. The reaction was monitored by TLC plate. DCM was removed by distillation under reduced pressure to obtain a white crude product, and column chromatography was performed using PE:EA=1:1 to obtain intermediate 3 (white solid, 3.56 g), with a yield of 87.73%.

[0051] Intermediate 3 (3.56 g, 13.32 mmol) was dissolved in DCM and stirred in an ice bath. A catalytic amount of p-toluenesulfonic acid (114.67 mg, 0.67 mmol) was added thereto, followed by slow dropwise addition of 3,4-dihydropyran (1.46 mL, 15.98 mmol) and continued stirring for 6 h. The reaction was monitored over a TLC plate, and DCM was removed under reduced pressure distillation to obtain a crude product. The crude product was purified by column chromatography with PE:EA=2:1 to obtain intermediate 4 (yellow oil, 2.72 g) with a yield of 58.12%.

[0052] Piperazine (800.08 mg, 9.28 mmol) was added to a DCM solution containing intermediate 4 (2.72 g, 7.74 mmol), HATU (3.24 g, 8.51 mmol) and DIPEA (4.04 mL, 23.22 mmol), stirred at room temperature for 30 min, and the reaction was monitored by TLC plate. An appropriate amount of water (200 mL) and DCM (200 mL×3) were added to the reaction solution for extraction, and the organic layers were combined, washed with saturated brine (400 mL), dried over anhydrous Na2SO4, and DCM was removed by vacuum distillation to obtain a pink oily crude product. The crude product was then purified by rapid column chromatography with DCM: MeOH = 10:1 to obtain the key intermediate 5 (yellow semi-solid, 1.83 g) with a yield of 56.35%.

[0053] Intermediate 5 (80 mg, 0.19 mmol) was dissolved in DMF, triethylamine (39.76 μL, 0.29 mmol) was added thereto and stirred at room temperature for 5 min. Acryloyl chloride (18.60 μL, 0.23 mmol) was slowly added to the mixture, and the reaction was carried out at room temperature for 30 min. The reaction was monitored by TLC plate to be complete. Water (50 mL) and DCM (50 mL×2) were added to the reaction solution, DCM was combined, and DMC was removed under vacuum to obtain a crude product. The crude product was dissolved in 6 mL of DCM, and 2 mL of TFA was added thereto, and the reaction was reacted at room temperature for 2 h. The reaction was monitored by TLC to be complete. The reaction solvent was spin-dried, adjusted to no bubbles with saturated sodium bicarbonate aqueous solution, and EA (50 mL×2) and water (50 mL) were added thereto for extraction, and the ethyl acetate phase was combined, washed with saturated brine (200 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to obtain a crude product. The crude product was purified by preparative TLC to give compound 1 (white solid, 52.23 mg) with a yield of 70.69%

[0054] Intermediate 1a (36.46 mg, 0.19 mmol) was added to DCM containing intermediate 5 (80 mg, 0.19 mmol), HATU (79.76 mg, 0.21 mmol) and DIPEA (99.7 μL, 0.57 mmol), and the reaction was carried out at room temperature for 30 min. The reaction was monitored by TLC. DCM (50 mL×3) and water (50 mL) were added thereto, the two DCMs were combined, and the DCM was removed under vacuum to obtain a crude product. The crude product was directly dissolved in DCM (6 mL), and 2 mL of TFA was added thereto, and the reaction was carried out at room temperature for 1 h to remove the THP protecting group. The solvent was removed by distillation under reduced pressure, and the system was adjusted to no bubbles using a saturated aqueous sodium bicarbonate solution. EA (50 mL×3) and water (50 mL) were added thereto for extraction, the ethyl acetate phase was combined, washed with saturated brine (100 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to obtain a crude product. The crude product was purified by preparative TLC (DCM:MeOH=12:1) to give compound 2 (white solid, 66.5 mg).

[0055] Compound 3-4 was synthesized via a synthetic route similar to that of compound 2.

[0056] 2. General Synthesis of Compound 5-13

[0057] The intermediate 6a-6j, i.e. aminobenzoic acid (1 eq) containing the corresponding substituent and potassium carbonate (K2CO3, 2.5 eq), were added to DMF and stirred overnight under an ice bath. Acryloyl chloride (1.2 eq) was then slowly added dropwise to the reaction mixture and the reaction was stirred overnight. TLC monitored the reaction to be complete and vacuum concentrated to obtain a white solid. A small amount of water was added to the solid to dissolve it, and EA was used to extract and remove fat-soluble impurities. The water layer was collected and the pH was adjusted to 3-4. A white solid was precipitated, filtered, and dried to obtain intermediates 7a-7j.

[0058] The intermediate 7a (36.46 mg, 0.19 mmol) was added to DCM containing the intermediate 5 (80 mg, 0.19 mmol), HATU (79.76 mg, 0.21 mmol) and DIPEA (99.7 μL, 0.57 mmol), and the reaction was carried out at room temperature for 30 min. The reaction was monitored by TLC. DCM (50 mL×3) and water (50 mL) were added thereto, the two DCMs were combined, and the DCM was removed under vacuum to obtain a crude product. The crude product was directly dissolved in DCM (6 mL), and 2 mL of TFA was added thereto, and the reaction was carried out at room temperature for 1 h to remove the THP protecting group. The solvent was removed by distillation under reduced pressure, and the system was adjusted with a saturated aqueous sodium bicarbonate solution until no bubbles were generated. EA (50 mL×3) and water (50 mL) were added thereto for extraction, the ethyl acetate phase was combined, washed with saturated brine (100 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to obtain a crude product. The crude product was purified by preparative TLC (DCM:MeOH=12:1) to give compound 5 (white solid, yield 55.26%).

[0059] Compounds 6-13 were synthesized via a synthetic route similar to that of compound 5.

[0060] 3. General Synthesis of Compounds 14-25

[0061] 3-amino-4-chlorobenzoic acid (1eq) and potassium carbonate (K2CO3, 2.5eq) were added to DCM and stirred under an ice bath. The acyl chloride derivative (1.2eq) was slowly added dropwise to the reactant and the reaction was stirred overnight. The reaction was monitored by TLC plate and the DCM was spun dry using a rotary evaporator under vacuum to obtain a white solid. A small amount of water was added to the solid to dissolve it, and EA was added to extract and remove fat-soluble impurities. The water layer was collected and the pH was adjusted to 3-4. The white solid was precipitated, filtered, and dried to obtain intermediates 8a-h with good purity.

[0062] Intermediate 8a-h (1 eq) was added to DCM containing intermediate 5 (1 eq), HATU (1.1 eq) and DIPEA (5.2 eq), and the mixture was reacted at room temperature for 30 min. The reaction was completed by monitoring by TLC. DCM (50 mL×3) and water (50 mL) were added thereto, the two DCMs were combined, and the DCM was removed under vacuum to obtain a crude product. The crude product was directly dissolved in DCM (6 mL), and 2 mL of TFA was added thereto. The reaction was reacted at room temperature for 1 h to remove the THP protecting group. The solvent was removed by distillation under reduced pressure, and the system was adjusted with saturated aqueous sodium bicarbonate solution until no bubbles were generated. EA (50 mL×3) and water (50 mL) were added thereto for extraction, and the ethyl acetate phases were combined, washed with saturated brine (100 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to obtain a crude product. The crude product was purified by preparative TLC (DCM: MeOH = 12: 1) to obtain the target compound 14-21.

[0063] Substituted piperazine was added to the DCM solution containing intermediate 4, HATU and DIPEA, stirred at room temperature for 30 min, and the reaction was monitored by TLC. Appropriate amount of water (200 mL) and DCM (200 mL×3) were added to the reaction mixture for extraction, and the organic layers were combined, washed with saturated brine (400 mL), dried over anhydrous Na2SO4, and DCM was removed by vacuum distillation to obtain an oily crude product. The crude product was then purified by column chromatography (DCM: MeOH = 10: 1) to obtain intermediates 9a-d.

[0064] Intermediate 6i was added to DCM containing intermediate 9a-d, HATU) and DIPE, and the reaction was carried out at room temperature for 30 minutes. The reaction was completed by TLC monitoring. DCM (50 mL×3) and water (50 mL) were added thereto, the two DCMs were combined, and the DCM was removed under vacuum to obtain a crude product. The crude product was directly dissolved in DCM (6 mL), and 2 mL of TFA was added thereto, and the reaction was carried out at room temperature for 1 hour to remove the THP protecting group. The solvent was removed by distillation under reduced pressure, and the system was adjusted with saturated sodium bicarbonate aqueous solution until no bubbles were generated. EA (50 mL×3) and water (50 mL) were added thereto for extraction, and the ethyl acetate phases were combined, washed with saturated brine (100 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to obtain a crude product. The crude product was purified by preparative TLC (DCM: MeOH = 12: 1) to obtain the target compounds 22-25.

[0065] 4. General Synthesis of Compounds 26-30

[0066] Tert-butyl 4-(chloromethyl)benzoate (1.1 eq) was added to 30 mL of DMF (30 mL) containing variously substituted 1H-indazole-3-amines (1 eq), KI (1 eq) and N,N-diisopropylethylamine (DIPEA, 2.7 eq), and the mixture was stirred at 70°C overnight. The reaction was monitored by TLC. EtOAc (125 mL) and water (100 mL) were added to the mixture, and the organic layer was separated. The aqueous layer was washed with EtOAc (125 mL×3), and the organic layers were combined and washed with water (100 mL×2) and brine (100 mL), dried over anhydrous Na2SO4, and concentrated in vacuo to give a crude product. The crude product was purified by column chromatography (PE:EtOAc=5:1, 4:1) to give intermediates 10a-e.

[0067] Intermediate 10a-e (1 eq) was dissolved in DCM (30 mL), and TFA (DCM:TFA=3:1) was added thereto, and the reaction was carried out at room temperature for 2-3 h. The reaction was monitored by TLC plate. DCM was removed by vacuum distillation to obtain a white crude product, which was purified by column chromatography using PE:EA=1:1 to obtain intermediate 11a-e.

[0068] Intermediate 11a-e (1 eq) was dissolved in DCM and stirred in an ice bath. A catalytic amount of p-toluenesulfonic acid (0.05 eq) was added, followed by slow dropwise addition of 3,4-dihydropyran (1.2 eq) and continued stirring for 6 h. The reaction was monitored using a TLC plate, and DCM was removed under reduced pressure distillation to obtain a crude product. The crude product was purified by column chromatography with PE:EA=2:1 to obtain intermediate 12a-e.

[0069] Piperazine (1.3 eq) was added to a DCM solution containing intermediate 12a-e (1 eq), HATU (3.24 g, 8.51 mmol) and DIPEA (1.1 eq), and stirred at room temperature for 30 min. The reaction was monitored by TLC plate. An appropriate amount of water (200 mL) and DCM (200 mL × 3) were added to the reaction solution for extraction. The organic layers were combined, washed with saturated brine (400 mL), dried over anhydrous Na2SO4, and DCM was removed by vacuum distillation to obtain a pink oily crude product. The crude product was then purified by rapid column chromatography with DCM: MeOH = 10:1 to obtain intermediate 13a-e.

[0070] Intermediate 7i (1 eq) was added to DCM containing intermediate 13a-e (1 eq), HATU (1.1 eq) and DIPEA (3 eq), and the reaction was carried out at room temperature for 30 min. The reaction was completed by TLC monitoring. DCM (50 mL×3) and water (50 mL) were added thereto, the two DCMs were combined, and the DCM was removed under vacuum to obtain a crude product. The crude product was directly dissolved in DCM (6 mL), and 2 mL of TFA was added thereto, and the reaction was carried out at room temperature for 1 h to remove the THP protecting group. The solvent was removed by distillation under reduced pressure, and the system was adjusted with saturated sodium bicarbonate aqueous solution until no bubbles were generated. EA (50 mL×3) and water (50 mL) were added thereto for extraction, and the ethyl acetate phases were combined, washed with saturated brine (100 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to obtain a crude product. The crude product was purified by preparative TLC (DCM: MeOH = 12: 1) to obtain the target compounds 26-30.

[0071] Compound 1, white solid, yield 70.69%; 1 H NMR (400MHz, DMSO-d6) δ11.85 (s, 1H), 7.48 (dd, J=8.1, 1.7Hz, 2H), 7.37 (dd, J=8.1, 1.8Hz, 2H), 7.24-7.18 (m, 2H), 6.96-6.89 (m, 1H) , 6.78 (s, 1H), 6.13 (dt, J=16.7, 2.0Hz, 1H), 6.07 (t, J=6.1Hz, 1H), 5.70 (dt, J=10.4, 2.0Hz, 1H), 4.54 (d, J=6.0Hz, 2H), 3.58 (s, 8H). 13 C NMR (101MHz, DMSO) δ169.75, 164.84, 149.02, 143.51, 143.29, 134.11, 128.49, 12 8.16, 127.91, 127.62, 127.47, 125.74, 118.31, 110.85, 109.26, 46.90.HRMS(ESI) + calculated for C 22 H 23 ClN5O2, [M+H] + : m / z424.1535, found 424.1542.

[0072] Compound 2, white solid, yield 66.62%; 1H NMR (400MHz, DMSO-d6) δ11.84 (s, 1H), 10.30 (s, 1H), 7.80 (s, 1H), 7.68 (dt, J=8.1, 1.5Hz, 1H), 7.4 8(d, J=7.8Hz, 2H), 7.41 (d, J=7.8Hz, 1H), 7.38 (d, J=7.7Hz, 2H), 7.24-7.17 (m, 2H), 7.12 (d, J=7.6 Hz, 1H), 6.92 (dd, J=5.3, 2.8Hz, 1H), 6.44 (dd, J=17.0, 10.0Hz, 1H), 6.28 (dd, J=17.0, 2.0Hz, 1H), 6.06 (t, J=6.1Hz, 1H), 5.85-5.70 (m, 1H), 4.54 (d, J=6.1Hz, 2H), 3.53 (d, J=85.8Hz, 8H).HRMS (ESI) + calculated for C 29 H 28 ClN6O3, [M+H] + : m / z 543.1906, found 543.1917.

[0073] Compound 3, white solid, yield 61.08%; 1 H NMR (400MHz, DMSO-d6) δ11.85 (d, J=2.9Hz, 1H), 10.35 (s, 1H), 7.80-7.63 (m, 2H), 7.48 ( d, J=7.8Hz, 2H), 7.41 (d, J=8.2Hz, 2H), 7.37 (d, J=7.7Hz, 2H), 7.24-7.17 (m, 2H), 6.92 (d d, J=5.3, 2.7Hz, 1H), 6.46 (ddd, J=16.9, 10.1, 2.8Hz, 1H), 6.29 (dd, J=16.9, 2.1Hz, 1H), 6.07 (t, J=6.1Hz, 1H), 5.79 (dd, J=10.1, 2.0Hz, 1H), 4.54 (d, J=6.1Hz, 2H), 3.53 (s, 8H). 13 C NMR (101MHz, DMSO) δ169.79, 169.47, 163.85, 149.01, 143.51, 143.29, 140.80, 134.07, 132.11, 13 0.62, 128.69, 127.92, 127.62, 127.50, 125.73, 119.26, 118.31, 110.83, 109.26, 46.89.HRMS(ESI) +calculated for C 29 H 28 ClN6O3, [M+H] + : m / z 543.1906, found 543.1910.

[0074] Compound 4, white solid, yield 65.76%; 1 H NMR (400MHz, DMSO-d6) δ11.84 (s, 1H), 10.47 (s, 1H), 7.96 (s, 1H), 7.59 (d, J=8.4Hz, 1 H), 7.47 (d, J=7.8Hz, 2H), 7.37 (d, J=7.4Hz, 2H), 7.34 (s, 1H), 7.24-7.15 (m, 2H), 6.9 6-6.88 (m, 1H), 6.43 (dd, J=16.9, 10.0Hz, 1H), 6.36-6.26 (m, 1H), 6.06 (t, J=6.1Hz, 1 H), 5.90-5.72 (m, 1H), 4.54 (d, J=6.1Hz, 2H), 3.68 (s, 2H), 3.49 (s, 4H), 3.22 (s, 2H). 13 C NMR (101MHz, DMSO) δ169.80, 166.15, 164.04, 149.02, 143.51, 143.32, 141.07, 133.99, 131.81, 130.58, 129.89 , 129.00, 128.39, 127.91, 127.61, 127.53, 125.73, 119.74, 118.49, 118.30, 110.84, 109.25, 46.89.HRMS (ESI) + calculated for C 29 H 27 Cl2N6O3, [M+H] + : m / z 577.1516, found 577.1520

[0075] Compound 5, white solid, yield 55.26%; 1H NMR (400MHz, DMSO-d6) δ11.84 (s, 1H), 9.55 (d, J = 8.6Hz, 1H), 7.58 (d, J = 46.7Hz, 1H), 7.47 (d, J = 7.7Hz, 2H), 7.37 (d, J=7.7Hz, 2H), 7.30 (d, J=7.3Hz, 1H), 7.20 (dd, J=4.0, 1.8Hz, 2H), 7.16 (t, J=6.8Hz, 1H), 6. 92 (dt, J=5.6, 1.9Hz, 1H), 6.56 (dd, J=17.0, 10.2Hz, 1H), 6.26 (d, J=17.0Hz, 1H), 6.06 (t, J=6.2Hz, 1H) , 5.77 (dt, J=10.1, 1.7Hz, 1H), 4.54 (d, J=6.1Hz, 2H), 3.54 (d, J=53.7Hz, 8H), 2.25 (d, J=4.5Hz, 3H).13C NMR (101MHz, DMSO) δ169.80, 169.30, 168.66, 163.88, 149.03, 143.51, 143.28, 136.44, 134.10, 133.58, 132.06, 130.92, 130.85, 127.90, 127.62, 127.49, 125.74, 124.34, 124.16, 123.80, 118.30, 110.85, 109.26, 46.92, 41.50, 18.30.HRMS(ESI)+calculated for C30H30ClN6O3, [M+H]+: m / z 557.2062, found 557.2061.

[0076] Compound 6, white solid, yield 49.93%; 11H NMR (400 MHz, DMSO-d6) δ 11.84 (s, 1H), 10.19 (s, 1H), 7.58 (s, 1H), 7.51 (dd, J = 8.4, 2.2 Hz, 1H), 7.46 (d, J = 7.8 Hz, 2H), 7.37 (d, J = 7.8 Hz, 2H), 7.23 (s, 1H), 7.21 - 7.17 (m, 2H), 6.92 (dd, J = 5.4, 2.7 Hz, 1H), 6.41 (dd, J = 16.9, 10.0 Hz, 1H), 6.25 (dd, J = 16.9, 2.1 Hz, 1H), 6.05 (s, 1H), 5.75 (dd, J = 10.0, 2.0 Hz, 1H), 4.53 (d, J = 6.1 Hz, 2H), 3.56 (d, J = 108.7 Hz, 6H), 3.27 - 3.18 (m, 2H), 2.17 (s, 3H). HRMS (ESI)+ calculated for C30H30ClN6O3, [M + H]+: m / z 557.2062, found 557.2062.

[0077] Compound 7, white solid, yield 57.56%; 1 1H NMR (400 MHz, DMSO-d6) δ 11.86 (s, 1H), 9.52 (s, 1H), 8.31 - 8.07 (m, 1H), 7.48 (d, J = 7.9 Hz, 2H), 7.38 (d, J = 7.9 Hz, 2H), 7.26 - 7.17 (m, 3H), 7.11 (d, J = 8.5 Hz, 1H), 6.92 (dd, J = 5.6, 2.4 Hz, 1H), 6.72 (dd, J = 16.9, 10.2 Hz, 1H), 6.25 (dd, J = 16.9, 2.0 Hz, 1H), 6.06 (t, J = 6.1 Hz, 1H), 5.76 - 5.72 (m, 2H), 4.54 (d, J = 6.1 Hz, 2H), 3.89 (s, 3H), 3.56 (s, 8H). 13C NMR (101 MHz, DMSO) δ 169.82, 169.51, 164.01, 151.12, 149.02, 143.51, 143.26, 134.10, 132.28, 127.91, 127.62, 127.49, 127.20, 125.74, 124.56, 121.66, 118.31, 111.30, 110.84, 109.25, 56.42, 55.36, 46.91. HRMS (ESI)+ calculated for C30H30ClN6O4, [M + H]+: m / z 573.2012, found 573.2012

[0078] Compound 8, white solid, yield 54.48%; 1 H NMR (400MHz, DMSO-d6) δ11.84 (s, 1H), 10.10 (s, 1H), 8.15 (d, J=7.4Hz, 1H), 7.47 (d, J=7 .8Hz, 2H), 7.37 (d, J=8.2Hz, 3H), 7.26-7.22 (m, 1H), 7.22-7.17 (m, 2H), 6.92 (dd, J=5.5 , 2.6Hz, 1H), 6.64 (dd, J=17.0, 10.2Hz, 1H), 6.29 (dd, J=17.0, 1.9Hz, 1H), 6.06 (t, J=6. 1Hz, 1H), 5.80 (dd, J=10.1, 2.0Hz, 1H), 4.54 (d, J=6.1Hz, 2H), 3.62 (s, 4H), 3.46 (s, 4H). 13 C NMR (101MHz, DMSO) δ169.83, 168.63, 164.17, 149.02, 143.51, 143.29, 134.06, 132.16, 132.13, 131.59, 128.35, 127.93, 12 7.63, 127.49, 126.60, 126.48, 125.74, 124.77, 124.70, 123.21, 118.32, 116.25, 116.06, 110.84, 109.26, 46.90.HRMS(ESI) + calculated for C 29 H 27 ClFN6O3, [M+H] + : m / z 561.1812, found561.1807.

[0079] Compound 9, white solid, yield 56.82%; 11H NMR (400 MHz, DMSO-d6) δ 11.84 (s, 1H), 10.49 (s, 1H), 7.68 (d, J = 11.2 Hz, 1H), 7.48 (s, 1H), 7.46 (s, 2H), 7.37 (d, J = 7.7 Hz, 2H), 7.25 - 7.16 (m, 2H), 6.99 (d, J = 8.5 Hz, 1H), 6.92 (dd, J = 5.5, 2.6 Hz, 1H), 6.41 (dd, J = 17.0, 10.0 Hz, 1H), 6.30 (dd, J = 17.0, 2.1 Hz, 1H), 6.06 (t, J = 6.1 Hz, 1H), 5.82 (dd, J = 9.9, 2.1 Hz, 1H), 4.54 (d, J = 6.1 Hz, 2H), 3.52 (d, J = 90.2 Hz, 8H). 13 13C NMR (101 MHz, DMSO) δ 169.80, 168.01, 164.05, 163.41, 160.99, 149.01, 143.51, 143.32, 141.35, 141.24, 138.41, 138.33, 134.04, 131.80, 128.41, 127.90, 127.62, 127.50, 125.73, 118.30, 114.16, 110.84, 109.25, 109.03, 107.66, 107.39, 46.90. HRMS (ESI) + calculated for C 29 H 27 ClFN6O3, [M + H] + : m / z 561.1812, found 561.1812.

[0080] Compound 10, white solid, yield 60.37%; 1 1H NMR (400 MHz, DMSO-d6) δ 11.85 (s, 1H), 9.79 (s, 1H), 7.76 (d, J = 8.5 Hz, 2H), 7.48 (d, J = 7.8 Hz, 2H), 7.37 (d, J = 7.8 Hz, 2H), 7.23 - 7.20 (m, 2H), 7.20 - 7.17 (m, 1H), 6.92 (dd, J = 5.6, 2.5 Hz, 1H), 6.62 (dd, J = 17.0, 10.2 Hz, 1H), 6.30 (dd, J = 16.8, 1.8 Hz, 1H), 6.06 (t, J = 6.1 Hz, 1H), 5.81 (dd, J = 10.1, 1.9 Hz, 1H), 4.55 (d, J = 6.1 Hz, 2H), 3.64 (s, 4H), 3.44 (s, 4H).13 C NMR (101MHz, DMSO) δ169.80, 168.27, 164.06, 149.02, 143.51, 143.30, 136.51, 135.56, 134.06, 133.43, 131.63 ,128.25,127.90,127.63,127.49,126.00,125.80,125.74,119.23,118.30,110.86,109.25,46.92.HRMS(ESI) + calculated for C 29 H 27 ClBrN6O3, [M+H] + : m / z 621.1011, found 621.1007.

[0081] Compound 11, white solid, yield 58.15%; 1 H NMR (400MHz, DMSO-d6) δ11.86 (s, 1H), 10.50 (s, 1H), 7.78 (s, 1H), 7.70-7.64 (m, 1 H), 7.49 (s, 1H), 7.49-7.45 (m, 2H), 7.38 (d, J=7.7Hz, 2H), 7.25-7.17 (m, 2H), 6.9 2 (dd, J=5.6, 2.5Hz, 1H), 6.45 (dd, J=17.0, 10.1Hz, 1H), 6.35-6.24 (m, 1H), 6.13- 5.98 (m, 1H), 5.84-5.74 (m, 1H), 4.54 (d, J=6.1Hz, 2H), 3.70 (s, 6H), 3.24 (s, 2H). 13 C NMR (101MHz, DMSO) δ169.83, 166.01, 163.91, 149.01, 143.51, 143.32, 138.83, 136.05, 133.97, 131.90, 130.35 ,128.09,127.91,127.62,127.53,125.73,123.45,121.62,118.62,118.31,110.83,109.26,46.89.HRMS(ESI) + calculated for C 29 H 27 Cl2N6O3, [M+H] + : m / z 577.1516, found 577.1517.

[0082] Compound 12, white solid, yield 62.74%; 1 H NMR (400MHz, DMSO-d6) δ11.84 (s, 1H), 10.44 (d, J = 3.9Hz, 1H), 8.18 (d, J = 8.2Hz, 1H), 7.87 (t, J=2.2Hz, 1H), 7.68 (t, J=8.1Hz, 1H), 7.61 (d, J=10.3Hz, 1H), 7.58-7.54 (m, 1H), 7.52 (s, 1H), 7 .47 (s, 1H), 7.38 (d, J=7.1Hz, 2H), 7.26-7.14 (m, 2H), 6.41 (dd, J=16.6, 8.9Hz, 1H), 6.30 (d, J =17.0Hz, 1H), 5.82 (dd, J=9.9, 2.1Hz, 1H), 4.60-4.53 (m, 2H), 3.79-3.59 (m, 4H), 3.54 (s, 4H). 13 C NMR (101MHz, DMSO) δ169.80, 167.85, 153.15, 143.51, 143.31, 142.02, 140.93, 139.23, 138.35, 133.75 ,132.79,131.77,128.45,127.91,127.42,126.65,121.89,120.11,114.91,110.84,46.49.HRMS(ESI) + calculated for C 29 H 27 Cl2N6O3, [M+H] + : m / z 577.1516, found 577.1517.

[0083] Compound 13, white solid, yield 70.56%; 11H NMR (400 MHz, DMSO-d6) δ 11.84 (s, 1H), 9.85 (s, 1H), 7.93 (d, J = 18.9 Hz, 1H), 7.59 (d, J = 8.2 Hz, 1H), 7.47 (d, J = 7.8 Hz, 2H), 7.37 (d, J = 7.7 Hz, 2H), 7.25 (d, J = 8.1 Hz, 1H), 7.22 - 7.15 (m, 2H), 6.92 (dt, J = 4.5, 2.1 Hz, 1H), 6.65 (dd, J = 17.1, 10.3 Hz, 1H), 6.29 (d, J = 17.0 Hz, 1H), 6.06 (t, J = 6.1 Hz, 1H), 5.81 (dd, J = 10.3, 2.1 Hz, 1H), 4.54 (d, J = 6.0 Hz, 2H), 3.63 (s, 4H), 3.44 (s, 4H). 13 13C NMR (101 MHz, DMSO) δ 169.81, 168.30, 164.14, 149.02, 143.51, 143.30, 135.15, 134.99, 134.07, 131.62, 130.20, 128.35, 127.90, 127.63, 127.49, 125.74, 125.35, 124.92, 118.30, 110.85, 109.25, 107.45, 46.91. HRMS (ESI) + calculated for C 29 H 27 Cl2N6O3, [M + H] + : m / z 577.1516, found 577.1520.

[0084] Compound 14, white solid, yield 70.56%; 1 1H NMR (400 MHz, DMSO-d6) δ 11.84 (s, 1H), 9.85 (s, 1H), 7.93 (d, J = 18.9 Hz, 1H), 7.59 (d, J = 8.2 Hz, 1H), 7.47 (d, J = 7.8 Hz, 2H), 7.37 (d, J = 7.7 Hz, 2H), 7.25 (d, J = 8.1 Hz, 1H), 7.22 - 7.15 (m, 2H), 6.92 (dt, J = 4.5, 2.1 Hz, 1H), 6.65 (dd, J = 17.1, 10.3 Hz, 1H), 6.29 (d, J = 17.0 Hz, 1H), 6.06 (t, J = 6.1 Hz, 1H), 5.81 (dd, J = 10.3, 2.1 Hz, 1H), 4.54 (d, J = 6.0 Hz, 2H), 3.63 (s, 4H), 3.44 (s, 4H).13 C NMR (101MHz, DMSO) δ169.81, 168.30, 164.14, 149.02, 143.51, 143.30, 135.15, 134.99, 134.07, 131.62, 130.20 ,128.35,127.90,127.63,127.49,125.74,125.35,124.92,118.30,110.85,109.25,107.45,46.91.HRMS(ESI) + calculated for C 29 H 27 Cl2N6O3, [M+H] + : m / z 577.1516, found 577.1520.

[0085] Compound 15, light yellow solid, yield 49.72%; 1 H NMR (400MHz, DMSO-d6) δ11.83 (s, 1H), 9.43 (s, 1H), 7.87 (s, 1H), 7.59-7.52 (m, 1H), 7.47 (d, J=7.7Hz, 2H), 7.39-7.34 (m, 2H), 7.24-7. 16 (m, 3H), 6.92 (dd, J=5.3, 2.8Hz, 1H), 6.07 (d, J=7.6Hz, 2H), 4.54 (d, J=5.8Hz, 2H), 3.53 (d, J=77.2Hz, 8H), 2.13 (s, 3H), 1.87 (s, 3H). 13 C NMR (101MHz, DMSO) δ169.78, 168.41, 167.70, 165.21, 153.32, 149.02, 143.51, 143.29, 135.62, 134.86, 13408, 13 0.01, 127.90, 127.62, 127.49, 125.73, 124.66, 118.90, 118.30, 110.85, 109.25, 46.90, 27.55, 20.14.HRMS(ESI) + calculated for C 31 H 31 Cl2N6O3, [M+H] + : m / z 605.1829, found 605.1832.

[0086] Compound 16, light yellow solid, yield 47.85%; 1H NMR (400MHz, DMSO-d6) δ11.84 (s, 1H), 9.54 (s, 1H), 7.65 (s, 1H), 7.60 (d, J=8.2Hz , 1H), 7.47 (d, J=7.8Hz, 2H), 7.37 (d, J=7.8Hz, 2H), 7.30 (dd, J=8.3, 2.0Hz, 1H), 7. 23-7.17 (m, 2H), 6.92 (dd, J=5.3, 2.8Hz, 1H), 6.06 (s, 1H), 5.91 (s, 1H), 5.61-5.5 5 (m, 1H), 4.54 (d, J = 6.1Hz, 2H), 3.63 (s, 4H), 3.43 (s, 4H), 1.98 (d, J = 10.1Hz, 3H). 13 C NMR (101MHz, DMSO) δ169.80, 168.20, 166.98, 149.02, 143.51, 143.31, 139.89, 135.41, 135.04, 134.06, 130.25, 13 0.13, 127.91, 127.63, 127.49, 126.53, 126.02, 125.73, 121.55, 118.30, 110.85, 109.25, 46.90, 19.00.HRMS(ESI) + calculated for C 30 H 29 Cl2N6O3, [M+H] + : m / z 591.1673, found 591.1676.

[0087] Compound 17, white solid, yield 62.15%; 1 H NMR (400MHz, DMSO-d6) δ11.84 (s, 1H), 9.98 (s, 1H), 7.94-7.74 (m, 1H), 7.60 (d, J = 8.2Hz, 1H), 7.47 (d, J = 7.8Hz, 2H), 7.37 (d, J = 7.8Hz, 2H), 7.28 (dd, J=8.3, 2.0Hz, 1H), 7.24-7.15(m, 2H), 6.92(dd, J=5.4, 2.7Hz, 1H), 6.06(t , J=6.1Hz, 1H), 4.54 (d, J=6.1Hz, 2H), 4.40 (s, 2H), 3.52 (d, J=77.0Hz, 8H). 13CNMR (101MHz, DMSO) δ169.81, 168.19, 165.90, 149.02, 146.23, 143.51, 143.31, 135.16, 134.72, 134.06, 1 30.27, 127.90, 127.63, 127.48, 125.74, 125.69, 124.73, 118.30, 110.85, 109.25, 46.91, 43.51.HRMS(ESI) + calculated for C 28 H 26 Cl3N6O3, [M+H] + : m / z 599.1126, found 599.1130.

[0088] Compound 18, white solid, yield 61.98%; 1 H NMR (400MHz, DMSO-d6) δ11.84 (s, 1H), 9.98 (s, 1H), 7.83 (s, 1H), 7.60 (d, J = 8.3Hz, 1H), 7.47 (d, J = 7.8Hz, 2H), 7.37 (d, J = 7.7Hz, 2H), 7.28 (d, J=8.4Hz, 1H), 7.24-7.17 (m, 2H), 6.92 (dd, J=5.4, 2.7Hz, 1H), 6.06 (s, 1H), 4.53 (d, J=4.6Hz, 2H), 4.40 (s, 2H), 3.52 (d, J=79.7Hz, 8H). 13 C NMR (101MHz, DMSO) δ169.80, 168.18, 165.90, 149.02, 147.14, 143.51, 143.31, 135.17, 134.72, 134.06, 13 0.27, 127.90, 127.63, 127.48, 125.73, 125.69, 124.73, 118.30, 110.85, 109.25, 46.91, 43.51.HRMS (ESI) + calculated for C 28 H 26 BrCl2N6O3, [M+H] + : m / z 643.0621, found643.0620.

[0089] Compound 19, white solid, yield 56.75%; 1H NMR (400MHz, DMSO-d6) δ11.85 (s, 1H), 10.44 (s, 1H), 7.80-7.71 (m, 1H), 7.64 (d, J=8.2Hz, 1H), 7.48 (d, J=7.8Hz, 2H), 7.39 (s, 1H), 7.38-7.31 (m, 2H ), 7.22(s, 1H), 7.21-7.17(m, 1H), 6.92(dd, J=5.8, 2.3Hz, 1H), 6.86(s, 1H ), 6.06 (t, J=6.1Hz, 1H), 4.55 (d, J=6.1Hz, 2H), 3.64 (s, 4H), 3.43 (s, 4H). 13 C NMR (101MHz, DMSO) δ169.82, 167.98, 163.15, 149.02, 143.51, 143.31, 135.38, 134.06, 133.95, 130.48, 12 8.98, 127.90, 127.64, 127.49, 126.62, 125.74, 125.41, 118.30, 110.85, 109.26, 67.13, 46.91.HRMS(ESI) + calculated for C 28 H 25 Cl4N6O3, [M+H] + : m / z 633.0737, found633.0734.

[0090] Compound 20, white solid, yield 64.12%; 1 H NMR (400MHz, DMSO-d6) δ11.84 (s, 1H), 10.88 (s, 1H), 7.66 (d, J=8.2Hz, 1H), 7.53-7.45 (m, 3H), 7.44-7.40 (m, 1H), 7.37 (d, J=7.8Hz, 2H), 7 .23-7.15 (m, 2H), 6.92 (dd, J=5.4, 2.7Hz, 1H), 6.06 (t, J=6.1Hz, 1H), 4.54 (d, J=6.1Hz, 2H), 3.53 (d, J=85.4Hz, 8H), 3.17 (d, J=5.2Hz, 2H). 13C NMR (101MHz, DMSO) δ169.81, 167.78, 160.94, 149.01, 143.51, 143.33, 135.55, 134.27, 134.04, 132.39, 13 0.58, 129.32, 127.98, 127.91, 127.64, 127.49, 125.73, 118.30, 110.84, 109.26, 93.04, 46.90.HRMS (ESI) + calculated for C 28 H 24 Cl5N6O3, [M+H] + : m / z 667.0347, found 667.0344.

[0091] Compound 21, white solid, yield 71.24%; 1 H NMR (400MHz, DMSO-d6) δ11.84 (s, 1H), 9.54 (s, 1H), 7.83 (s, 1H), 7.55 (d, J=8. 2Hz, 1H), 7.47 (d, J=7.8Hz, 2H), 7.37 (d, J=7.7Hz, 2H), 7.21 (q, J=4.6, 3.5Hz, 3H), 6.92 (dd, J=5.7, 2.4Hz, 1H), 6.06 (t, J=6.1Hz, 1H), 4.54 (d, J=6.0Hz, 2H) , 3.62 (s, 4H), 3.55-3.39 (m, 4H), 2.42 (d, J = 7.6Hz, 2H), 1.09 (t, J = 7.5Hz, 3H). 13 C NMR (101MHz, DMSO) δ173.05, 169.83, 168.40, 149.02, 143.51, 143.30, 137.75, 135.49, 134.91, 134.04, 132.8 0, 130.08, 127.93, 127.63, 127.49, 125.73, 124.94, 118.32, 110.83, 109.26, 46.89, 29.43, 10.09.HRMS(ESI) + calculated for C 29 H 29 Cl2N6O3, [M+H] + : m / z 579.1673, found 579.1674.

[0092] Compound 22, white solid, yield 65.28%; 1H NMR (400MHz, DMSO-d6) δ11.84 (s, 1H), 9.84 (s, 1H), 7.90 (s, 1H), 7.59 (d, J = 8.2Hz, 1H), 7.47 (d, J = 7.8Hz, 2H), 7. 35-7.33 (m, 2H), 7.21 (s, 1H), 7.20 (d, J=2.8Hz, 1H), 6.92 (d, J=2.7Hz, 1H), 6.91 (d, J=2.6Hz, 1H), 6.65 (dd, J=17 .0, 10.2Hz, 1H), 6.29 (dd, J=17.0, 1.9Hz, 1H), 6.06 (t, J=6.1Hz, 1H), 5.81 (dd, J=10.2, 1.9Hz, 1H), 4.54 (d, J=6. 1Hz, 2H), 4.35-4.23 (m, 1H), 3.52 (d, J=82.7Hz, 2H), 3.18 (s, 2H), 2.99 (d, J=28.9Hz, 2H), 1.17 (t, J=7.1Hz, 3H). 13 C NMR (101MHz, DMSO) δ164.15, 154.88, 154.58, 149.04, 147.73, 143.52, 143.12, 135.19, 134.52, 133.83, 131.62, 13 0.29, 128.63, 128.38, 127.91, 127.70, 127.11, 125.74, 125.30, 118.31, 110.84, 109.26, 50.26, 46.89.HRMS(ESI) + calculated for C 30 H 29 Cl2N6O3, [M+H] + : m / z 591.1673, found591.1674.

[0093] Compound 23, white solid, yield 66.43%; 1H NMR (400MHz, DMSO-d6) δ11.85 (s, 1H), 9.84 (s, 1H), 7.88 (s, 1H), 7.59 (d, J = 8.2Hz, 1H), 7.47 (d, J = 7.9Hz, 2H ), 7.36 (d, J = 7.7Hz, 2H), 7.25-7.22 (m, 1H), 7.21 (s, 1H), 7.20 (d, J = 3.1Hz, 1H), 6.92 (dd, J = 5.6, 2.6Hz, 1H), 6.65 (dd, J=17.0, 10.2Hz, 1H), 6.29 (dd, J=17.0, 2.0Hz, 1H), 6.06 (d, J=6.2Hz, 1H), 5.81 (dd, J=10.2, 1.9Hz, 1H), 4.54 (d, J=6.1Hz, 2H), 4.28 (s, 1H), 3.57 (s, 2H), 3.15 (s, 2H), 3.03-2.84 (m, 2H), 1.17 (t, J=7.1Hz, 3H). 13 C NMR (101MHz, DMSO) δ169.88, 164.14, 150.61, 149.03, 143.51, 143.22, 135.43, 134.09, 131.64, 130.28, 128.37, 12 7.91, 127.67, 127.42, 126.35, 125.74, 124.52, 119.36, 118.30, 110.84, 109.26, 107.78, 46.91, 45.27.HRMS (ESI) + calculated for C 30 H 29 Cl2N6O3, [M+H] + : m / z 591.1673, found 591.1678.

[0094] Compound 24, white solid, yield 67.13%; 1H NMR (400MHz, DMSO-d6) δ11.85 (s, 1H), 9.85 (s, 1H), 7.89 (s, 1H), 7.59 (d, J = 8.2Hz, 1H), 7.47 (d, J = 7.8Hz , 2H), 7.34 (d, J=7.6Hz, 2H), 7.23 (d, J=6.1Hz, 1H), 7.21 (t, J=2.9Hz, 2H), 6.91 (dd, J=5.6, 2.6Hz, 1H), 6 .69-6.61 (m, 1H), 6.29 (d, J = 16.8Hz, 1H), 6.06 (t, J = 6.0Hz, 1H), 5.81 (d, J = 10.2Hz, 1H), 4.54 (d, J = 6.1H z, 2H), 4.28 (s, 1H), 3.51 (d, J = 86.2Hz, 2H), 3.19 (s, 2H), 2.99 (d, J = 44.9Hz, 2H), 1.24 (d, J = 6.6Hz, 3H). 13 C NMR (101MHz, DMSO) δ164.16, 149.03, 145.10, 143.51, 143.11, 135.17, 134.99, 134.75, 134.49, 131.81, 131.60, 130.2 8, 128.41, 127.93, 127.70, 127.08, 125.73, 125.31, 124.73, 118.31, 110.83, 109.26, 46.89, 46.13, 34.54.HRMS(ESI) + calculated for C 30 H 29 Cl2N6O3, [M+H] + : m / z 591.1673, found 591.1677.

[0095] Compound 25, white solid, yield 61.96%; 1H NMR (400MHz, DMSO-d6) δ11.85 (s, 1H), 9.85 (s, 1H), 7.88 (s, 1H), 7.59 (d, J = 8.2Hz, 1H), 7.47 (d, J = 7.8Hz, 2 H), 7.35 (t, J=7.2Hz, 2H), 7.28-7.22 (m, 1H), 7.20 (dd, J=5.4, 1.7Hz, 2H), 6.95-6.88 (m, 1H), 6.66 (dd, J=17 .0, 10.2Hz, 1H), 6.30 (dd, J=16.9, 2.0Hz, 1H), 6.06 (t, J=6.1Hz, 1H), 5.81 (dd, J=10.2, 1.9Hz, 1H), 4.54 (d , J=6.1Hz, 2H), 4.29 (s, 1H), 3.59 (d, J=17.5Hz, 2H), 3.16 (s, 2H), 2.95 (d, J=6.3Hz, 2H), 1.21-1.08 (m, 3H). 13 C NMR (101MHz, DMSO) δ168.37, 164.12, 149.03, 143.51, 143.22, 135.44, 135.22, 134.52, 134.10, 131.63, 130.27, 128.98, 128.35, 127.90, 127.66, 127.57, 127.41, 127.06, 125.73, 118.30, 110.84, 109.25, 46.91, 45.97.HRMS(ESI)+calculated for C 30 H 29 Cl2N6O3, [M+H]+: m / z 591.1673, found 591.1676.

[0096] Compound 26, white solid, yield 47.16%; 1H NMR(400MHz,DMSO-d6)δ11.86-11.69(m,1H),9.82(d,J=19.2Hz,1H),7.86(d,J=32.2Hz,1H),7.58(t,J=7.2Hz,1H),7.47(d,J=7.6Hz,2H),7.36(d,J=7.9Hz,2H),7.33-7.10(m,2H),7.05(dd,J=8.4,1.9Hz,1H),6.67(d,J=10.7Hz,1H),6.65-6.56(m,1H),6.29(dd,J=16.4,2.2Hz,1H),6.27-6.09(m,1H),5.82-5.69(m,1H),4.48(d,J=6.0Hz,2H),3.60(s,4H),3.42(s,4H). 13 C NMR(101MHz,DMSO)δ169.84,168.32,164.16,157.75,155.28,147.96,145.03,144.94,143.32,135.14,134.97,134.03,131.59,130.21,128.39,128.18,128.09,127.72,127.43,125.38,124.93,106.41,103.29,103.08,102.60,102.42,46.87,42.30.HRMS(ESI)+calculated forC 29 H 27 ClFN6O3,[M+H]+:m / z 561.1812,found 561.1812.

[0097] Compound 27, white solid, yield 48.17%; 1H NMR (400 MHz, DMSO-d6) δ 11.49 (s, 1H), 10.04 (s, 1H), 8.38 (s, 1H), 8.20 (s, 1H), 7.77 (dd, J = 8.8, 5.4 Hz, 1H), 7.58 (d, J = 8.2 Hz, 1H), 7.46 (d, J = 7.8 Hz, 3H), 7.37 (d, J=7.8Hz, 3H), 7.20 (d, J=8.4Hz, 1H), 6.98 (dd, J=10.1, 2.2Hz, 1H), 6.80-6.74 (m, 1H), 6.71 (d, J=6.1Hz, 1H), 4.49 (d, J=6.0Hz, 2H), 3.62 (s, 4H), 3.43 (s, 4H)..13C NMR (101MHz, DMSO) δ169.79, 168.31, 163.59, 161.20, 150.09, 143.21, 142.33, 135.25, 134.17, 130.22, 130.12, 127.79, 127.50, 124.94, 124.34, 122.34, 122.23, 121.99, 111.27, 107.24, 106.98, 95.37, 95.11, 46.72.HRMS(ESI)+calculated forC29H27ClFN6O3, [M+H]+: m / z 561.1812, found 561.1814.

[0098] Compound 28, white solid, yield 47.16%; 1 H NMR (400MHz, DMSO-d6) δ12.07 (s, 1H), 9.83 (d, J=19.7Hz, 1H), 7.82 (d, J=62.5H z, 1H), 7.59 (dd, J=8.8, 3.8Hz, 2H), 7.46 (d, J=7.8Hz, 2H), 7.38 (dd, J=8.1, 1.9 Hz, 2H), 7.25 (d, J=8.1Hz, 1H), 7.09-6.90 (m, 2H), 6.90-6.82 (m, 1H), 6.29 (d, J =17.1Hz, 1H), 5.81 (dd, J = 10.2, 1.9Hz, 1H), 4.50 (d, J = 6.0Hz, 2H), 3.59 (s, 8H). 13C NMR (101MHz, DMSO) δ169.78, 169.09, 164.16, 154.90, 150.66, 149.32, 142.95, 135.56, 135.14, 134.97, 134.26, 131.61, 129.85, 1 28.68, 127.81, 127.78, 127.56, 127.53, 124.93, 121.28, 119.35, 117.02, 114.51, 108.46, 108.25, 46.76.HRMS(ESI)+calculated for C 29 H 26 ClF2N6O3, [M+H]+: m / z 579.1717, found 579.1722

[0099] Compound 29, white solid, yield 49.78%; 1 H NMR (400MHz, DMSO-d6) δ11.38 (s, 1H), 9.82 (d, J = 19.1Hz, 1H), 7.86 (d, J = 32.4Hz, 1H), 7.75 (d, J=7.9Hz, 1H), 7.58 (t, J=7.2Hz, 1H), 7.47 (d, J=7.8Hz, 2H), 7.37 (d, J=7.8Hz, 2H), 7.32-7.13 (m, 3H), 6.90 (ddd, J=7.9, 4.6, 3.1Hz, 1H), 6.69-6.56 (m, 1H), 6.36-6. 07 (m, 1H), 5.81 (dd, J=10.2, 1.9Hz, 1H), 4.51 (d, J=6.1Hz, 2H), 3.51 (d, J=60.4Hz, 8H). 13 C NMR (101MHz, DMSO) δ169.82, 168.31, 164.15, 150.11, 143.43, 142.24, 135.13, 134.97, 134.11, 133.78, 131.60, 1 30.22, 127.79, 127.48, 126.79, 125.39, 124.93, 120.49, 117.88, 114.05, 109.82, 46.82.HRMS(ESI)+calculated for C 29 H 28 ClN6O3, [M+H]+: m / z 543.1906, found 543.1909.

[0100] Compound 30, white solid, yield 50.16%;

[0101] 1 H NMR (400MHz, DMSO-d6) δ11.61 (s, 1H), 9.85 (s, 1H), 7.91 (s, 1H), 7.86 (d, J= 1.9Hz, 1H), 7.59 (d, J=8.1Hz, 1H), 7.47 (d, J=7.8Hz, 2H), 7.38 (d, J=7.8Hz, 2H), 7.28-7.22(m, 3H), 6.71-6.60(m, 2H), 6.29(dd, J=17.0, 2.0Hz, 1H), 5. 81 (dd, J=10.1, 1.9Hz, 1H), 4.51 (d, J=6.0Hz, 2H), 3.63 (s, 4H), 3.45 (s, 4H). 13 C NMR (101MHz, DMSO) δ169.79, 168.32, 164.16, 149.63, 143.08, 140.62, 135.13, 134.97, 134.21, 131.60, 130.22, 128. 39, 127.78, 127.52, 126.97, 125.39, 124.94, 121.96, 119.87, 114.73, 111.54, 49.07, 46.75.HRMS(ESI)+calculated for C 29 H 27 Cl2N6O3, [M+H]+: m / z 577.1516, found577.1516.

[0102] Test Example 1 Kinase inhibitory activity of compounds 1 to 30

[0103] The purpose of this experiment is to detect the inhibitory activity of the compounds of the present invention on JNK1 and JNK3 enzymes in vitro. The results are shown in Tables 1, 2 and 3.

[0104] Table 1. JNK1 and JNK3 inhibitory activity test results of compounds related to Formula I

[0105]

[0106]

[0107] Table 2. JNK1 and JNK3 inhibitory activity test results of compounds related to Formula II

[0108]

[0109]

[0110] Table 3. JNK1 and JNK3 inhibitory activity test results of compounds related to Formula III

[0111]

[0112]

[0113] The experimental results showed that the preferred compounds had strong inhibitory activity against JNK3, and the inhibitory activity of each compound was good, among which the optimal compound 13 had the strongest inhibitory effect.

[0114] Experimental Example 2 In vitro study on the protective activity of compound 13 on nerve cells

[0115] The purpose of this experiment is to detect the in vitro protective activity of the inventive compound on nerve cells. This experiment uses the MPP+ induced Parkinson's model, the cell line used is SY5Y, and the protective activity of the inventive compound 13 is tested.

[0116] SH-SY5Y cells were seeded in a medium with 5×10 3 In a 96-well plate with 10 cells / well, the cells were cultured for 24 hours. To study the protective activity of compound 13 on SH-SY5Y cells, the cells were pretreated with compound 13 for 8 hours and then exposed to 1 mM MPP+. After 24 hours of treatment, the corresponding OD value was measured using an ELISA instrument, and the protective activity was evaluated using the MTT method.

[0117] Results Figure 1 AB, from Figure 1 It can be clearly seen from AB that the compound has a significant dose-dependent protective activity against SY5Y cells.

[0118] Experimental Example 3 Neuroprotective effect of compound 13 on in vivo PD model

[0119] The purpose of this experiment is to detect the in vivo neuroprotective potential of the inventive compound 13. We used the neurotoxin MPTP to induce motor dysfunction in PD mice and constructed an MPTP-induced PD mouse model.

[0120] In this study, mice were placed in Plexiglas cages and placed under illumination with a light intensity of 50-65 lux. Spontaneous locomotor activity was tested for 60 minutes using the Photobeam Activity System-Open Field. The total number of beams blocked by each mouse was measured as an indicator of locomotor activity.

[0121] Results Figure 1 C, from Figure 1 It can be clearly seen in C that compound 13 can dose-dependently improve the motor function deficits induced by MPTP in mice.

[0122] The above experiments show that compound 13 can play a good neuroprotective effect in vitro and in vivo by selectively inhibiting JNK3, providing a good method for the treatment of Parkinson's disease.

Claims

1. A compound having a structural formula as shown in Formula I, II, III and IV or a pharmaceutically acceptable salt thereof: in, R 1 for R 2 for R 3 for 2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The structural formulas are shown in Formulas I, II, III and IV.

3. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, characterized in that: R 1 for R 2 for R 3 for 4. Use of the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof in the preparation of an anti-Parkinson's drug.

5. The use according to claim 4, characterized in that: The anti-Parkinson's drug is a covalent inhibitor drug targeting JNK3.

6. A pharmaceutical composition, characterized in that: The invention relates to a preparation comprising an effective dose of the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof.

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