A pyrazolo[3,4-d]pyrimidin-4-one derivative and use thereof

Abstract

The application belongs to the technical field of pharmaceutical chemistry, and discloses a pyrazolo[3,4-d]pyrimidine-4-ketone derivative or a pharmaceutically acceptable salt thereof with the following structure, which is used for preventing or treating pain diseases, R1 represents a substituent selected from hydrogen, halogen or alkyl; m is an integer of 3-5; n is an integer of 0-2; X is a carbon atom, an oxygen atom or a nitrogen atom; and R2 represents a substituent selected from hydrogen, a phenyl group, a substituted or unsubstituted alkyl group.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of pharmaceutical chemistry, and particularly relates to a pyrazolo[3,4-d]pyrimidine-4-ketone derivative and application thereof in treating pain diseases. BACKGROUND

[0002] Pain is one of the most common symptoms in clinic. According to the pathological characteristics, pain is divided into nociceptive pain and neuropathic pain. Nociceptive pain is a reaction caused by nociceptors sensing harmful stimuli and is related to tissue damage. When nerve fibers are damaged or the nervous system is damaged or diseased abnormally, spontaneous impulses are generated, causing pain at the origin of the nerve, which is called neuropathic pain (NP) and is also commonly referred to as neuralgia. However, existing analgesic drugs for neuropathic pain still have problems such as limited analgesic efficiency, low response rate, and different degrees of side effects and adverse reactions, especially when pregabalin is used in combination with high-dose opioid drugs, it may increase respiratory depression and even death.

[0003] σ1 receptors are widely expressed in multiple regions of the nervous system and are involved in the regulation of neuropathic pain and the regulation of neuroimmune cell activities including microglia, thereby affecting the inflammatory response process. Subsequent experiments have proved that σ1 receptors act through direct interaction with opioid μ receptors. In the formalin-induced pain experiment using σ1 receptor knockout mice, there was no obvious pain response in phase I and phase II, and the same phenomenon occurred in the capsaicin-stimulated neuropathic pain model. In the paclitaxel-induced cold-induced pain hypersensitivity and mechanical stimulation pain hypersensitivity model and the sciatic nerve ligation neuropathic pain model, σ1 receptor knockout mice also showed pain insensitivity behavior. σ1 receptor antagonists are good anti-inflammatory analgesic targets, and on the other hand, σ1 receptor antagonists themselves have pain, especially neuropathic pain analgesic effect. In recent years, research institutions and pharmaceutical companies have shown great interest in the development of σ1 receptor antagonists. Currently, S1RA is used for the treatment of various pain experiments and enters the clinical stage, and is used alone for the treatment of neuropathic pain experiments have entered clinical research. When normal mice are subjected to pain model experiments with σ1 receptor antagonists, they show similar pain desensitization to σ1 receptor knockout mice. The above experiments prove that haloperidol and its metabolites with σ1 receptor antagonistic effect have obvious analgesic effect in various neuropathic pain models. Therefore, searching for selective σ1 receptor antagonists for anti-pain treatment has important scientific value and social significance for the clinical treatment of pain and neuropathic pain. SUMMARY

[0004] The present application aims at providing a pyrazolo[3,4-d]pyrimidin-4-one derivative and its application in treating pain-related diseases to solve the above problems existing in the prior art.

[0005] To achieve the above object, the technical scheme of the present application is as follows.

[0006] In a first aspect, the present application provides a pyrazolo[3,4-d]pyrimidin-4-one derivative, the structural general formula of which is as follows:

[0007]

[0008] In the above formula, R1 represents a group selected from hydrogen, halogen or alkyl; m is an integer of 3-5; n is an integer of 0-2; X is a carbon atom, an oxygen atom or a nitrogen atom; and R2 represents a group selected from hydrogen, phenyl, substituted or unsubstituted alkyl.

[0009] The alkyl is methyl.

[0010] The halogen is fluorine.

[0011] The pyrazolo[3,4-d]pyrimidin-4-one derivative is selected from any one of the following compounds:

[0012] 5-{[3-(hexahydropyrrol-1-yl)propyl]oxy}-6-methyl-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one;

[0013] 6-methyl-5-{[3-(1,4-oxazinane-4-yl)propyl]oxy}-1-phenyl-4,5-dihydro-pyrazolo[3,4-d]pyrimidin-4-one;

[0014] 6-methyl-5-{[3-(4-methylpiperazin-1-yl)propyl]oxy}-1-phenyl-4,5-dihydro-1H-pyrazolo[3,4-d]pyrimidin-4-one;

[0015] 6-methyl-1-phenyl-5-{[3-(4-phenylpiperazin-1-yl)propyl]oxy}-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one;

[0016] 6-methyl-1-phenyl-5-{[3-(tetrahydro-1H-pyrrol-1-yl)propyl]oxy}-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one;

[0017] 5-{[3-(azolidin-1-yl)propyl]oxy}-6-methyl-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one;

[0018] 6-methyl-5-{[2-(1,4-oxazepan-4-yl)ethyl]oxy}-1-phenyl-4,5-dihydropyrazolo[3,4- d]pyrimidin-4-one;

[0019] 6-methyl-5-{[4-(1,4-oxazepan-4-yl)butyl]oxy}-1-phenyl-4,5-dihydropyrazolo[3,4- d]pyrimidin-4-one;

[0020] 6-methyl-5-{[4-(4-methylpiperazin-1-yl)butyl]oxy}-1-phenyl-4,5-dihydropyrazolo[3,4- d]pyrimidin-4-one;

[0021] 6-methyl-1-phenyl-5-{[4-(4-phenylpiperazin-1-yl)butyl]oxy}-4,5-dihydropyrazolo[3,4- d]pyrimidin-4-one;

[0022] 5-{[5-(hexahydropyridin-1-yl)pentyl]oxy}-6-methyl-1-phenyl-4,5-dihydropyrazolo[3,4- d]pyrimidin-4-one;

[0023] 6-methyl-5-{[5-(1,4-oxazepan-4-yl)pentyl]oxy}-1-phenyl-4,5-dihydropyrazolo[3,4- d]pyrimidin-4-one;

[0024] 6-methyl-5-{[5-(4-methylpiperazin-1-yl)pentyl]oxy}-1-phenyl-4,5-dihydropyrazolo[3,4- d]pyrimidin-4-one;

[0025] 6-methyl-1-phenyl-5-{[5-(4-phenylpiperazin-1-yl)pentyl]oxy}-4,5-dihydropyrazolo[3,4- d]pyrimidin-4-one;

[0026] 6-methyl-1-phenyl-5-{[5-(tetrahydro-1H-pyrrol-1-yl)pentyl]oxy}-4,5-dihydro-1H- pyrazolo[3,4-d]pyrimidin-4-one;

[0027] 1-(4-fluorophenyl)-5-{[3-(hexahydropyridin-1-yl)propyl]oxy}-6-methyl-4,5-dihydro-1H- pyrazolo[3,4-d]pyrimidin-4-one;

[0028] 1-(4-fluorophenyl)-6-methyl-5-{[3-(1,4-oxazepan-4-yl)propyl]oxy}-4,5-dihydro-1H- pyrazolo[3,4-d]pyrimidin-4-one;

[0029] 1 -(3,4-dimethylphenyl)-6-methyl-5-{[3-(1,4-oxazepan-4- yl)propyl]oxy}-4,5-dihydro-1 H-pyrazolo[3,4-d]pyrimidin-4-one;

[0030] 1 -(3,4-dimethylphenyl)-6-methyl-5-{[3-(1,4-oxazepan-4- yl)propyl]oxy}-4,5-dihydro-1 H-pyrazolo[3,4-d]pyrimidin-4-one;

[0031] 1 -(3,4-dimethylphenyl)-6-methyl-5-{[3-(1,4-oxazepan-4- yl)propyl]oxy}-4,5-dihydro-1 H-pyrazolo[3,4-d]pyrimidin-4-one;

[0032] 1 -(3,4-dimethylphenyl)-6-methyl-5-{[3-(1,4-oxazepan-4- yl)propyl]oxy}-4,5-dihydro-1 H-pyrazolo[3,4-d]pyrimidin-4-one;

[0033] 1 -(3,4-dimethylphenyl)-6-methyl-5-{[3-(1,4-oxazepan-4- yl)propyl]oxy}-4,5-dihydro-1 H-pyrazolo[3,4-d]pyrimidin-4-one;

[0034] 1 -(3,4-dimethylphenyl)-6-methyl-5-{[3-(1,4-oxazepan-4- yl)propyl]oxy}-4,5-dihydro-1 H-pyrazolo[3,4-d]pyrimidin-4-one;

[0035] 1 -(3,4-dimethylphenyl)-6-methyl-5-{[3-(1,4-oxazepan-4- yl)propyl]oxy}-4,5-dihydro-1 H-pyrazolo[3,4-d]pyrimidin-4-one;

[0036] 1 -(3,4-dimethylphenyl)-6-methyl-5-{[3-(1,4-oxazepan-4- yl)propyl]oxy}-4,5-dihydro-1 H-pyrazolo[3,4-d]pyrimidin-4-one;

[0037] 1 -(3,4-dimethylphenyl)-6-methyl-5-{[3-(1,4-oxazepan-4- yl)propyl]oxy}-4,5-dihydro-1 H-pyrazolo[3,4-d]pyrimidin-4-one.

[0038] In a second aspect, the present application provides a pharmaceutical composition comprising a therapeutically effective amount of the compound or a pharmaceutically acceptable salt thereof as described above, and a pharmaceutically acceptable excipient.

[0039] The salt is a pharmaceutically acceptable anion salt, including hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate or bisulfate, phosphate or acid phosphate, acetate, lactate, citrate, tartrate, maleate, fumarate, methanesulfonate, gluconate, saccharate, benzoate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate.

[0040] The pharmaceutically acceptable adjuvant includes carriers, excipients, disintegrants, sweeteners, lubricants, etc.; the carrier includes inert solid fillers, diluents, sterile aqueous or organic solutions; for the convenience of oral treatment, the target product or its pharmaceutically acceptable salt is used together with excipients to be made into tablets, lozenges, capsules, suspensions, syrups, etc.

[0041] The therapeutically effective amount depends on the type and severity of the disease or condition, and also depends on the characteristics of the subject, such as general health, age, sex, body weight, and drug tolerance. A person skilled in the art can determine the appropriate dose according to these or other factors. The effective dose of the central nervous system drugs commonly used is well known to the skilled person, and the total daily dose is usually between 0.05 mg and 2000 mg.

[0042] The pharmaceutical composition can be administered by any appropriate route, such as oral administration in the form of capsules, parenteral administration in the form of injection solutions, topical administration in the form of ointments or lotions, rectal administration in the form of suppositories, and transdermal administration in the form of patch delivery systems.

[0043] In a third aspect, the present application provides the use of the aforementioned pharmaceutical composition in the prevention or treatment of pain-related diseases.

[0044] The pain-related diseases refer to acute pain (such as acute injury pain of soft tissue and joints, postoperative pain, obstetric pain, acute herpes zoster pain, gout, etc.), chronic pain (such as strain or degenerative pain of soft tissue and joints, intervertebral disc-derived pain, neurogenic pain, etc.), intractable pain (such as trigeminal neuralgia, post-herpetic neuralgia, intervertebral disc herniation, intractable headache, etc.), cancer pain (such as advanced tumor pain, tumor metastasis pain, etc.), and special pain (such as thrombotic vasculitis, intractable angina pectoris, idiopathic chest and abdominal pain, etc.).

[0045] Compared with the prior art, the present application has the following beneficial effects:

[0046] The present application provides a pyrazolo[3,4-d]pyrimidine-4-ketone derivative, and the compound is proved to have selective antagonism to sigma 1 receptor through in vitro receptor binding test, and the compound can obviously improve the formalin-induced phase I and phase II pain through animal test results, since these in vitro action targets and in vivo pharmacological models are closely related to the response of sigma 1 receptor-mediated nervous system regulation, especially pain, so the compound has the effect of treating pain, especially neuropathic pain, and provides a new path for the preparation of a drug for preventing or treating pain diseases. BRIEF DESCRIPTION OF DRAWINGS

[0047] Figure 1 Results of formalin-induced phase I pain experiment in mice.

[0048] Figure 2 Results of formalin-induced phase II pain experiment in mice. DETAILED DESCRIPTION

[0049] The present application will be further described in detail below in combination with specific embodiments and drawings.

[0050] The general synthetic route of the pyrazolo[3,4-d]pyrimidine-4-ketone derivative provided by the present application is to synthesize a pyrazolo[3,4-d]pyrimidine-4-ketone parent compound first, and then link it with a carbon chain and a nitrogen-containing structure.

[0051] Example 1:

[0052] This example provides 5-{[3-(hexahydro-pyran-1-yl)propyl]oxy}-6-methyl-1-phenyl-4,5-dihydro-pyrazolo[3,4-d]pyrimidine-4-ketone, and the synthesis steps are as follows:

[0053] The first stage, as Figure 1The first stage, as shown, ethyl (2E)-2-cyano-3-ethoxyprop-2-enoate and phenyl ethyl azide are dissolved in anhydrous ethanol solution, heated by reflux for 5 hours to obtain ethyl 5-amino-1-phenylpyrazole-4-carboxylate, and then ethyl 5-amino-1-phenylpyrazole-4-carboxylate is mixed with sodium hydroxide in methanol, and heated by reflux condensation to obtain 5-amino-1-phenylpyrazole-4-carboxylic acid, which is then mixed with acetic anhydride by condensation reflux, and heated for 4 hours to obtain 6-methyl-1-phenyl-4H-pyrazolo[3,4-d][1,3]oxazin-4-one, which is finally mixed with hydroxylamine hydrochloride in dry pyridine, and heated by reflux for 8 hours to obtain the first stage intermediate compound 5-hydroxy-6-methyl-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one; the reaction formula of the first stage is as follows:

[0054]

[0055] The second stage, referring to the synthesis method of Williamson ether, the first stage intermediate compound is mixed with 1,3-dibromopropane in an acetone solution with anhydrous potassium carbonate as a catalyst, heated by reflux at 60°C oil bath for 5-8 hours, and the molar ratio of the feeding substances is about 1:2 to obtain the first stage intermediate compound 5-[(3-bromopropyl)oxy]-6-methyl-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one; the reaction formula of the second stage is as follows:

[0056]

[0057] The third stage, the second stage intermediate compound is mixed with piperidine to obtain a reaction product; the reaction formula of the third stage is as follows:

[0058]

[0059] Nuclear magnetic resonance hydrogen spectrum: 1H NMR (600 MHz, DMSO-d6) δ 8.37 (s, 1H), 8.02 (dd, J = 8.7, 1.1 Hz, 2H), 7.58 (dd, J = 8.6, 7.5 Hz, 2H), 7.42 (t, J = 7.4 Hz, 1H), 4.28 (s, 2H), 3.29 (s, 2H), 2.64 (s, 3H), 2.48 - 2.24 (m, 4H), 1.91 (s, 2H), 1.52 (s, 4H), 1.40 (s, 2H). Nuclear magnetic resonance carbon spectrum: 13C NMR (176 MHz, DMSO-d6) δ 158.46, 53.77, 149.48, 138.57, 136.60, 129.78, 127.71, 122.25, 107.62, 74.96, 54.85, 54.33, 25.99, 25.39, 24.51, 20.75. MS (ESI) m / z 368.26 ([M+H]+).

[0060] Example 2:

[0061] This example provides 6-methyl-5-{[3-(1,4-oxazinane-4-yl)propyl]oxy}-1-phenyl-4,5- dihydro-pyrazolo[3,4-d]pyrimidin-4-one, the synthesis of which is the same as in Example 1, except that piperidine is replaced by morpholine.

[0062] Nuclear magnetic resonance hydrogen spectrum: 1H NMR (600 MHz, DMSO-d6) δ 8.37 (s, 1H), 8.02 (dd, J = 8.7, 1.1 Hz, 2H), 7.58 (dd, J = 8.6, 7.5 Hz, 2H), 7.42 (t, J = 7.4 Hz, 1H), 4.28 (s, 2H), 3.29 (s, 2H), 2.64 (s, 3H), 2.48 - 2.24 (m, 4H), 1.91 (s, 2H), 1.52 (s, 4H), 1.40 (s, 2H). Nuclear magnetic resonance carbon spectrum: 13C NMR (176 MHz, DMSO-d6) δ 158.46, 53.77, 149.48, 138.57, 136.60, 129.78, 127.71, 122.25, 107.62, 74.96, 54.85, 54.33, 25.99, 25.39, 24.51, 20.75. MS (ESI) m / z 368.26 ([M+H]+).

[0063] Example 3:

[0064] This example provides 6-methyl-5-{[3-(4-methylpiperazin-1-yl)propyl]oxy}-1- phenyl-4,5-dihydro-1H-pyrazolo[3,4-d]pyrimidin-4-one, the synthesis of which follows the procedure of Example 1, except that piperidine is replaced by 1- methylpiperazine.

[0065] Nuclear magnetic resonance hydrogen spectrum:1H NMR (600 MHz, DMSO-d6) δ 8.36 (s, 1H), 8.02 (dd, J = 8.7, 1.1 Hz, 2H), 7.57 (dd, J = 8.6, 7.5 Hz, 2H), 7.42 (t, J = 7.4 Hz, 1H), 4.26 (t, J = 6.2 Hz, 2H), 3.29 (s, 2H), 2.63 (s, 3H), 2.27 (s, 11H), 1.91 (s, 2H). Nuclear magnetic resonance carbon spectrum:13C NMR (176 MHz, DMSO-d6) δ 158.46, 153.76, 149.48, 138.57, 136.61, 129.78, 127.72, 122.25, 107.62, 74.86, 54.77, 53.98, 52.37, 45.53, 25.35, 20.80. MS (ESI) m / z 383.27 ([M+H]+: 383.22).

[0066] Example 4:

[0067] This example provides 6-methyl-1-phenyl-5-{[3-(4-phenylpiperazin-1-yl)propyl]oxy}- 4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one, the synthesis of which follows the procedure of Example 1, except that piperidine is replaced by 1-phenylpiperazine.

[0068] Nuclear magnetic resonance hydrogen spectrum: 1H NMR (600 MHz, DMSO-d6) δ 8.36 (s, 1H), 8.02 (dd, J = 8.7, 1.1 Hz, 2H), 7.57 (dd, J = 8.6, 7.5 Hz, 2H), 7.41 (t, J = 7.4 Hz, 1H), 7.21 (dd, J = 8.7, 7.3 Hz, 2H), 6.93 (d, J = 7.7 Hz, 2H), 6.77 (t, J = 7.3 Hz, 1H), 4.30 (t, J = 6.2 Hz, 2H), 3.29 (d, J = 1.1 Hz, 2H), 3.16 - 3.11 (m, 4H), 2.65 (s, 3H), 2.55 (s, 4H), 1.96 (p, J = 6.7 Hz, 2H). Nuclear magnetic resonance carbon spectrum: 13C NMR (176 MHz, DMSO-d6) δ 158.48, 153.77, 151.54, 149.48, 138.57, 136.61, 129.77, 129.41, 127.71, 122.25, 119.27, 115.82, 107.63, 74.94, 54.22, 53.11, 48.69, 25.43, 20.82. MS (ESI) m / z 445.30 ([M+H]+: 445.23).

[0069] Example 5:

[0070] This example provides 6-methyl-1-phenyl-5-{[3-(tetrahydro-1H-pyrrol-1- yl)propyl]oxy}-4,5-dihydropyrrolo[3,4-d]pyrimidin-4-one, the synthesis of which is the same as Example 1, except that piperidine is replaced by pyrrolidine.

[0071] Nuclear magnetic resonance hydrogen spectrum: 1H NMR (600 MHz, DMSO-d6) δ 8.36 (s, 1H), 8.02 (dd, J = 8.7, 1.1 Hz, 2H), 7.57 (dd, J = 8.6, 7.5 Hz, 2H), 7.41 (t, J = 7.4 Hz, 1H), 7.21 (dd, J = 8.7, 7.3 Hz, 2H), 6.93 (d, J = 7.7 Hz, 2H), 6.77 (t, J = 7.3 Hz, 1H), 4.30 (t, J = 6.2 Hz, 2H), 3.29 (d, J = 1.1 Hz, 2H), 3.16 - 3.11 (m, 4H), 2.65 (s, 3H), 2.55 (s, 4H), 1.96 (p, J = 6.7 Hz, 2H). Nuclear magnetic resonance carbon spectrum: 13C NMR (176 MHz, DMSO-d6) δ 158.48, 153.77, 151.54, 149.48, 138.57, 136.61, 129.77, 129.41, 127.71, 122.25, 119.27, 115.82, 107.63, 74.94, 54.22, 53.11, 48.69, 25.43, 20.82. MS (ESI) m / z 445.30 ([M+H]+: 445.23).

[0072] Example 6:

[0073] This example provides 5-{[3-(azetidin-l-yl)propyl]oxy}-6-methyl-l-phenyl-4,5- dihydropyr azo [3,4-d]pyrimidin-4-one, which is synthesized according to the procedure of Example 1, except that piperidine is replaced by cyclohexanecarboxamide.

[0074] Nuclear magnetic resonance hydrogen spectrum: 1H NMR (700 MHz, DMSO-d6) δ 8.37 (s, 1H), 8.10-7.98 (m, 2H), 7.64-7.54 (m, 2H), 7.48-7.38 (m, 1H), 4.28 (t, J = 5.9 Hz, 2H), 2.63 (s, 3H), 2.60 (s, 4H), 1.89 (s, 2H), 1.59 (s, 6H), 1.57-1.49 (m, 4H), 1.38-1.20 (m, 2H). Nuclear magnetic resonance carbon spectrum: 13C NMR (176 MHz, DMSO-d6) δ 158.41, 153.74, 149.46, 138.57, 136.60, 129.76, 127.69, 122.23, 107.63, 74.97, 65.52, 55.18, 53.93, 28.41, 26.99, 20.79. MS (ESI) m / z 382.28 ([M+H]+: 382.22).

[0075] Example 7:

[0076] This example provides 6-methyl-5-{[2-(l,4-oxazepan-4-yl)ethyl]oxy}-l-phenyl-4,5- dihydropyr azo [3,4-d]pyrimidin-4-one, which is synthesized according to the procedure of Example 1, except that the intermediate compound 5-hydroxy-6-methyl-l-phenyl-4,5- dihydropyr azo [3,4-d]pyrimidin-4-one of the first stage is reacted with 4-(2- chloroethyl)-l,4-oxazepane to synthesize the target product; the reaction formula is as follows:

[0077]

[0078] Nuclear magnetic resonance hydrogen spectrum: 1H NMR (700 MHz, DMSO-d6) δ 8.36 (s, 1H), 8.02 (d, J = 7.3 Hz, 2H), 7.58 (q, J = 8.3, 6.7 Hz, 2H), 7.42 (t, J = 7.2 Hz, 1H), 4.39 (t, J = 5.7 Hz, 2H), 3.57 (s, 4H), 2.84 - 2.67 (m, 4H), 2.66 (s, 3H), 2.49 (s, 2H). Nuclear magnetic resonance carbon spectrum: 13C NMR (176 MHz, DMSO-d6) δ 158.41, 153.74, 149.46, 138.57, 136.60, 129.76, 127.69, 122.23, 107.63, 74.97, 65.52, 55.18, 53.93, 28.41, 26.99, 20.79. MS (ESI) m / z 356.23 ([M+H]+: 356.17).

[0079] Example 8:

[0080] This example provides 6-methyl-5-{[4-(1,4-oxazepan-4-yl)butyl]oxy}-1-phenyl-4,5- dihydropyr azo lo [3,4-d]py rim id in-4-one, the synthesis steps of which are the same as in Example 1, except that 1,3-dibromopropane is replaced by 1,4-dibromobutane and piperidine is replaced by morpholine, to obtain the second-stage intermediate product as 5-[(4-bromobutyl)oxy]-6-methyl-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one.

[0081] Nuclear magnetic resonance hydrogen spectrum: 1H NMR (600 MHz, DMSO-d6) δ 8.37 (s, 1H), 8.02 (dd, J = 8.7, 1.1 Hz, 2H), 7.58 (dd, J = 8.6, 7.5 Hz, 2H), 7.42 (t, J = 7.4 Hz, 1H), 4.28 (s, 2H), 3.29 (s, 2H), 2.64 (s, 3H), 2.48 - 2.24 (m, 4H), 1.91 (s, 2H), 1.52 (s, 4H). Nuclear magnetic resonance carbon spectrum: 13C NMR (176 MHz, DMSO-d6) δ 158.46, 153.77, 149.48, 138.57, 136.60, 129.78, 127.71, 122.25, 107.62, 74.96, 54.85, 54.33, 25.99, 25.39, 24.51, 20.75. MS (ESI) m / z 368.26 ([M+H]+: 368.21).

[0082] Example 9:

[0083] This example provides 6-methyl-5-{[4-(4-methylpiperazin-1-yl)butyl]oxy}-1-phenyl- 4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one, the synthesis of which follows the procedure of Example 8, except that morpholine is replaced by 1-methylpiperazine.

[0084] Nuclear magnetic resonance hydrogen spectrum:1H NMR (600 MHz, DMSO-d6) δ 8.36 (s, 1H), 8.05-8.00 (m, 2H), 7.57 (dd, J = 8.6, 7.4 Hz, 2H), 7.42 (t, J = 7.4 Hz, 1H), 4.24 (t, J = 6.5 Hz, 2H), 3.58 (s, 4H), 3.30 (s, 2H), 2.62 (s, 3H), 2.37 (s, 4H), 1.80-1.73 (m, 2H), 1.67-1.59 (m, 2H). Nuclear magnetic resonance carbon spectrum:13C NMR (176 MHz, DMSO-d6) δ 158.41, 153.78, 149.46, 138.57, 136.60, 129.78, 127.71, 122.23, 107.63, 76.65, 57.70, 55.24, 53.09, 46.20, 25.95, 23.02, 20.93. MS (ESI) m / z 397.29 ([M+H]+: 397.23).

[0085] Example 10:

[0086] This example provides 6-methyl-1-phenyl-5-{[4-(4-phenylpiperazin-1-yl)butyl]oxy}-4,5- dihydropyrazolo[3,4-d]pyrimidin-4-one, the synthesis of which follows the procedure of Example 8, except that morpholine is replaced by 1-phenylpiperazine.

[0087] Nuclear magnetic resonance hydrogen spectrum: 1H NMR (600 MHz, DMSO-d6) δ 8.36 (s, 1H), 8.03 (dd, J = 8.7, 1.1 Hz, 2H), 7.57 (dd, J = 8.6, 7.5 Hz, 2H), 7.41 (t, J = 7.4 Hz, 1H), 7.20 (dd, J = 8.8, 7.3 Hz, 2H), 6.93 (d, J = 7.9 Hz, 2H), 6.77 (t, J = 7.3 Hz, 1H), 4.27 (t, J = 6.4 Hz, 2H), 3.13 (t, J = 4.9 Hz, 4H), 2.62 (s, 3H), 2.53 (s, 4H), 2.42 (s, 2H), 1.83 - 1.75 (m, 2H), 1.67 (p, J = 7.2 Hz, 2H). Nuclear magnetic resonance carbon spectrum: 13C NMR (176 MHz, DMSO-d6) δ 158.40, 153.75, 151.54, 149.46, 138.57, 136.60, 129.77, 129.40, 127.69, 122.21, 119.24, 115.82, 107.63, 76.64, 57.74, 53.22, 48.70, 25.99, 23.00, 20.94. MS (ESI) m / z 459.31 ([M+H]+: 459.25).

[0088] Example 11:

[0089] This example provides 5-{[5-(hexahydropyridin-l-yl)pentyl]oxy}-6-methyl-l- phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one, which is synthesized according to the procedure of Example 1, except that 1,3-dibromopropane is replaced by 1,5- dibromopentane, resulting in the second stage intermediate 5-[(5-bromopentyl)oxy]-6- methyl-l-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one.

[0090] Nuclear magnetic resonance hydrogen spectrum: 1H NMR (600 MHz, DMSO-d6) δ 8.36 (s, 1H), 8.02 (dd, J = 8.7, 1.1 Hz, 2H), 7.57 (dd, J = 8.6, 7.4 Hz, 2H), 7.41 (t, J = 7.4 Hz, 1H), 4.25-4.22 (m, 2H), 2.61 (s, 3H), 2.31 (s, 4H), 2.25 (t, J = 7.1 Hz, 2H), 1.75 (p, J = 6.7 Hz, 2H), 1.48 (q, J = 5.5 Hz, 8H), 1.37 (t, J = 7.5 Hz, 2H). Nuclear magnetic resonance carbon spectrum: 13C NMR (176 MHz, DMSO-d6) δ 158.40, 153.75, 149.46, 138.58, 136.59, 129.78, 127.70, 122.21, 107.63, 76.68, 58.96, 54.57, 27.97, 26.51, 26.04, 24.65, 23.81, 20.92. MS (ESI) m / z 396.29 ([M+H]+: 396.24).

[0091] Example 12:

[0092] This example provides 6-methyl-5-{[5-(1,4-oxazinane-4-yl)pentyl]oxy}-1-phenyl-4,5- dihydropyrazolo[3,4-d]pyrimidin-4-one, the synthesis of which is the same as Example 11, except that piperidine is replaced by morpholine.

[0093] Nuclear magnetic resonance hydrogen spectrum: 1H NMR (600 MHz, DMSO-d6) δ 8.36 (s, 1H), 8.02 (dd, J = 8.7, 1.2 Hz, 2H), 7.57 (dd, J = 8.6, 7.4 Hz, 2H), 7.42 (t, J = 7.4 Hz, 1H), 4.24 (t, J = 6.5 Hz, 2H), 3.58 (s, 4H), 3.30 (s, 2H), 2.61 (s, 3H), 2.36 (s, 4H), 1.82-1.71 (m, 2H), 1.51 (p, 4H). Nuclear magnetic resonance carbon spectrum: 13C NMR (176 MHz, DMSO-d6) δ 158.40, 153.75, 149.46, 138.57, 136.59, 129.78, 127.70, 122.22, 107.63, 76.64, 66.62, 58.56, 53.81, 27.92, 26.02, 23.65, 20.92. MS (ESI) m / z 398.27 ([M+H]+: 398.22).

[0094] Example 13:

[0095] This example provides 6-methyl-5-{[5-(4-methylpiperazin-1-yl)pentyl]oxy}-1- phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one, synthesized according to the procedure of Example 11, except that piperidine is replaced with 1- methylpiperazine.

[0096] Nuclear magnetic resonance hydrogen spectrum:1H NMR (600 MHz, DMSO-d6) δ 8.36 (s, 1H), 8.02 (dd, J = 8.7, 1.1 Hz, 2H), 7.57 (dd, J = 8.6, 7.5 Hz, 2H), 7.41 (t, J = 7.4 Hz, 1H), 4.25-4.22 (m, 2H), 2.61 (s, 3H), 2.31 (s, 4H), 2.25 (t, J = 7.1 Hz, 2H), 1.75 (p, J = 6.7 Hz, 2H), 1.48 (q, J = 5.5 Hz, 8H), 1.37 (t, J = 7.5 Hz, 2H). Nuclear magnetic resonance carbon spectrum:13C NMR (176 MHz, DMSO-d6) δ 158.40, 153.75, 149.46, 138.58, 136.59, 129.78, 127.70, 122.21, 107.63, 76.68, 58.96 54.57, 27.97, 26.51, 26.04, 24.65, 23.81, 20.92. MS (ESI) m / z 396.29 ([M+H]+: 396.24).

[0097] Example 14:

[0098] This example provides 6-methyl-1-phenyl-5-{[5-(4-phenylpiperazin-1-yl)pentyl]oxy}- 4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one, synthesized according to the procedure of Example 11, except that piperidine is replaced with 1-phenylpiperazine.

[0099] Nuclear magnetic resonance hydrogen spectrum: 1H NMR (600 MHz, DMSO-d6) δ 8.36 (s, 1H), 8.02 (dd, J = 8.7, 1.1 Hz, 2H), 7.57 (dd, J = 8.6, 7.5 Hz, 2H), 7.41 (t, J = 7.4 Hz, 1H), 7.20 (dd, J = 8.8, 7.2 Hz, 2H), 6.93 (d, J = 7.7 Hz, 2H), 6.77 (t, J = 7.2 Hz, 1H), 4.25 (t, J = 6.4 Hz, 2H), 3.13 (s, 4H), 2.62 (s, 3H), 2.58 - 2.51 (m, 4H), 2.37 (s, 2H), 1.78 (p, J = 6.7 Hz, 2H), 1.61 - 1.53 (m, 2H), 1.53 - 1.45 (m, 2H). Nuclear magnetic resonance carbon spectrum: 13C NMR (176 MHz, DMSO-d6) δ 158.41, 153.76, 151.48, 149.46, 138.57, 136.60, 129.78, 129.42, 127.71, 122.22, 119.29, 115.84, 107.64, 76.65, 58.17, 53.24, 48.62, 27.93, 26.45, 23.73 20.94. MS (ESI) m / z 473.32 ([M+H]+: 473.26).

[0100] Example 15:

[0101] This example provides 6-methyl-l-phenyl-5-{[5-(tetrahydro-lH-pyrrol-l- yl)pentyl]oxy}-4,5-dihydro-lH-pyrazolo[3,4-d]pyrimidin-4-one, synthesized according to the procedure of Example 11, except that piperidine is replaced by pyrrolidine.

[0102] Nuclear magnetic resonance hydrogen spectrum: 1H NMR (600 MHz, DMSO-d6) δ 8.36 (s, 1H), 8.02 (dd, J = 8.7, 1.1 Hz, 2H), 7.57 (dd, J = 8.6, 7.5 Hz, 2H), 7.42 (t, J = 7.4 Hz, 1H), 4.24 (t, J = 6.3 Hz, 2H), 2.93 (s, 4H), 2.85 (t, J = 7.8 Hz, 2H), 2.62 (s, 3H), 1.83 (p, J = 3.6 Hz, 4H), 1.78 (p, 2H), 1.66 (p, J = 7.7 Hz, 2H), 1.50 (p, J = 7.6 Hz, 2H). Nuclear magnetic resonance carbon spectrum: 13C NMR (176 MHz, DMSO-d6) δ 158.38, 153.75, 149.45, 138.56, 136.58, 129.78, 127.71, 122.21, 107.62, 76.49, 54.92, 53.77, 27.68, 26.60, 23.28, 21.66, 20.94. MS (ESI) m / z 382.28 ([M+H]+: 382.22).

[0103] Example 16:

[0104] This example provides 1 -(4-fluorophenyl)-5-{[3-(hexahydropyridin-1 -yl)propyl]oxy}- 6-methyl-4,5-dihydro-1 H-pyrazolo[3,4-d]pyrimidin-4-one, synthesized according to the procedure of Example 1, except that phenethyl azide is replaced with (4- fluorophenyl)ethyl azide, (2E)-2-cyano-3-ethoxyprop-2-enenitrile (1 ) is dissolved in anhydrous ethanol with (4-fluorophenyl)ethyl azide and the reaction is heated to reflux for 5 hours to give ethyl 5-amino-1 -(4-fluorophenyl)pyrazole-4-carboxylate, which is mixed with sodium hydroxide in methanol and the reaction is heated to reflux by condensation to hydrolyze to give 5-amino-1 -(4-fluorophenyl)pyrazole-4-carboxylic acid, which is then mixed with acetic anhydride by condensation to reflux and the reaction is heated for 4 hours, filtered, dried and crystallized to give 1 -(4-fluorophenyl)-6-methyl-4H-pyrazolo[3,4-d][1,3]oxazin-4-one, which is finally mixed with hydroxylamine hydrochloride in dry pyridine and the reaction is heated to reflux for 8 hours to give the first stage intermediate compound as 1 -(4-fluorophenyl)-5-hydroxy-6-methyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one; the second stage intermediate compound is 5-[(3-bromopropyl)oxy]-1 -(4-fluorophenyl)-6-methyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one.

[0105] Nuclear magnetic resonance hydrogen spectrum:1H NMR (600 MHz, DMSO-d6) δ 8.36 (s, 1 H), 8.04 (dd, J = 9.1, 4.9 Hz, 2H), 7.42 (t, J = 8.8 Hz, 2H), 4.25 (t, J = 6.2 Hz, 2H), 2.63 (s, 3H), 2.43 (t, 2H), 2.33 (s, 4H), 1.89 (p, J = 6.6 Hz, 2H), 1.49 (p, J = 5.7 Hz, 4H), 1.38 (s, 2H). Nuclear magnetic resonance carbon spectrum:13C NMR (176 MHz, DMSO-d6) δ 161.85, 160.46, 158.62, 153.72, 149.43, 136.64, 134.98, 134.96, 124.43, 124.38, 116.69, 116.56, 107.53, 75.01, 54.91, 54.41, 26.08, 25.48, 24.63, 20.73. MS (ESI) m / z 386.27 ([M+H]+: 386.20).

[0106] Example 17:

[0107] This example provides 1 -(4-fluorophenyl)-6-methyl-5-{[3-(1,4-oxazinane-4- yl)propyl]oxy}-4,5-dihydro-1 H-pyrazolo[3,4-d]pyrimidin-4-one, which is synthesized according to the procedure of Example 16, except that piperidine is replaced by morpholine.

[0108] Nuclear magnetic resonance hydrogen spectrum:1H NMR (600 MHz, DMSO-d6) δ 8.36 (s, 1 H), 8.04 (dd, J = 9.1, 4.9 Hz, 2H), 7.46 - 7.39 (m, 2H), 4.27 (t, J = 6.2 Hz, 2H), 3.58 (t, J = 4.6 Hz, 4H), 3.30 (s, 2H), 2.63 (s, 3H), 2.39 (s, 4H), 1.91 (p, J = 6.6 Hz, 2H). Nuclear magnetic resonance carbon spectrum:13C NMR (176 MHz, DMSO-d6) δ 161.84, 160.46, 158.58, 153.71, 149.41, 136.63, 134.97, 134.95, 124.40, 124.35, 116.69, 116.56, 107.52, 74.87, 66.68, 54.62, 53.66, 25.08 20.77. MS (ESI) m / z 388.24 (M+H]+388.18).

[0109] Example 18:

[0110] This example provides 1 -(4-fluorophenyl)-6-methyl-5-{[4-(1,4-oxazinane-4- yl)butyl]oxy}-4,5-dihydro-1 H-pyrazolo[3,4-d]pyrimidin-4-one, which is synthesized according to the procedure of Example 16, except that 1,3-dibromopropane is replaced by 1,4-dibromobutane and piperidine is replaced by morpholine, resulting in the second stage intermediate compound 5-[(4-bromobutyl)oxy]-1 -(4-fluorophenyl)-6-methyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one.

[0111] Nuclear magnetic resonance hydrogen spectrum:1H NMR (600 MHz, DMSO-d6) δ 8.37 (s, 1H), 8.06 (dd, J = 9.1, 4.9 Hz, 2H), 7.47-7.40 (m, 2H), 4.25 (t, J = 6.4 Hz, 2H), 3.59 (t, J = 4.6 Hz, 4H), 2.62 (s, 3H), 2.41-2.28 (m, 6H), 1.77 (p, 2H), 1.63 (p, J = 6.8 Hz, 2H). Nuclear magnetic resonance carbon spectrum:13C NMR (176 MHz, DMSO-d6) δ 161.84, 160.45, 158.53, 153.70, 149.39, 136.62, 134.97, 134.95, 124.37, 124.32, 116.69, 116.56, 107.53, 76.63 66.69, 58.14, 53.78, 25.90, 22.64, 20.92. MS (ESI) m / z 402.25 ([M+H]+: 402.19).

[0112] Example 19:

[0113] This example provides 1-(3,4-dimethylphenyl)-5-{[3-(hexahydropyridin-1-yl)propyl]oxy}-6-methyl-4,5-dihydro-1H-pyrazolo[3,4-d]pyrimidin-4-one. The synthesis steps are the same as those in Example 1, except that phenylethazane is replaced with 1-(3,4-dimethylphenyl)ethazane, (2E)-2-cyano-3-ethoxyprop-2-enoic acid ethyl ester (1) is reacted with 1-(3,4-dimethylphenyl)ethazane, and the reaction mixture is stirred for 2 h. 4-dimethylphenyl) ethazane is dissolved in anhydrous ethanol solution and refluxed to obtain 5-amino-1-(3,4-dimethylphenyl) pyrazole-4-carboxylic acid ethyl ester. 5-amino-1-(3,4-dimethylphenyl) pyrazole-4-carboxylic acid ethyl ester is mixed with sodium hydroxide in methanol and refluxed to obtain 5-amino-1-(3,4-dimethylphenyl) pyrazole-4-carboxylic acid. 5-amino-1-(3,4-dimethylphenyl) pyrazole-4-carboxylic acid ethyl ester is then hydrolyzed to obtain 5-amino-1-(3,4-dimethylphenyl) pyrazole-4-carboxylic acid. (3,4-Dimethylphenyl) pyrazole-4-carboxylic acid and acetic anhydride were mixed, condensed and refluxed, heated to react for 4 hours, filtered, dried and crystallized to obtain 1-(3,4-dimethylphenyl)-6-methyl-4H-pyrazolo[3,4-d][1,3]oxazahexacyclohexan-4-one. Finally, 1-(3,4-dimethylphenyl)-6-methyl-4H-pyrazolo[3,4-d][1,3]oxazahexacyclohexan-4-one was reacted with hydroxybenzoic acid hydrochloride. The amine is mixed in dry pyridine and heated under reflux for 8 hours to obtain the intermediate compound of the first stage, which is 1-(3,4-dimethylphenyl)-5-hydroxy-6-methyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one; the intermediate compound of the second stage is 5-[(3-bromopropyl)oxy]-1-(3,4-dimethylphenyl)-6-methyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one.

[0114] Nuclear magnetic resonance hydrogen spectrum: 1H NMR (600 MHz, DMSO-d6) δ 8.31 (s, 1H), 7.74 (s, 1H), 7.71 (d, J = 7.9 Hz, 1H), 7.31 (d, J = 8.1 Hz, 1H), 4.25 (t, J = 6.2 Hz, 2H), 2.62 (s, 3H), 2.43 (t, J = 7.0 Hz, 2H), 2.33 (s, 4H), 2.31 (s, 3H), 2.28 (s, 3H), 1.89 (p, J = 6.6 Hz, 2H), 1.49 (p, J = 5.5 Hz, 4H), 1.39 (p, J = 5.9 Hz, 2H). Nuclear magnetic resonance carbon spectrum: 13C NMR (176 MHz, DMSO-d6) δ 158.23, 153.78, 149.26, 137.77, 136.39, 136.20, 136.04, 130.48, 123.37, 119.83, 107.39, 74.97, 54.92, 54.41, 26.09, 25.49, 24.64, 20.75, 20.11, 19.49. MS (ESI) m / z 396.31 ([M+H]+: 396.24).

[0115] Example 20:

[0116] This example provides 1 -(3,4-dimethylphenyl)-6-methyl-5-{[3-(1,4-oxazepan-4- yl)propyl]oxy}-4,5-dihydro-1 H-pyrazolo[3,4-d]pyrimidin-4-one, synthesized according to the procedure of Example 19, except that piperidine is replaced by morpholine.

[0117] Nuclear magnetic resonance hydrogen spectrum: 1H NMR (600 MHz, DMSO-d6) δ 8.31 (s, 1H), 7.75 (d, J = 2.3 Hz, 1H), 7.71 (dd, J = 8.1, 2.3 Hz, 1H), 7.31 (d, J = 8.2 Hz, 1H), 4.27 (t, J = 6.3 Hz, 2H), 3.58 (t, J = 4.5 Hz, 4H), 3.30 (s, 2H), 2.62 (s, 3H), 2.38 (s, 4H), 2.31 (s, 3H), 2.28 (s, 3H), 1.91 (p, J = 6.5 Hz, 2H). Nuclear magnetic resonance carbon spectrum: 13C NMR (176 MHz, DMSO-d6) δ 158.23, 153.78, 149.26, 137.78, 136.38, 136.21, 136.06, 130.49, 123.38, 119.84, 107.38, 74.85, 66.71, 54.65, 53.69, 25.12, 20.80, 20.11, 19.49. MS (ESI) m / z 398.28 ([M+H]+: 398.22).

[0118] Example 21:

[0119] This example provides 1 -(3, 4-dimethylphenyl)-6-methyl-5-{[3-(4- phenylpiperazin-1 -yl)propyl]oxy}-4, 5-dihydro-1 H-pyrazolo[3, 4-d]pyrimidin-4-one, the synthesis of which is the same as Example 19, except that piperidine is replaced by 1 - phenylpiperazine.

[0120] Nuclear magnetic resonance hydrogen spectrum: 1H NMR (600MHz DMSO-d6) δ 8.31 (s, 1H), 7.75 (d, J = 2.3 Hz, 1H), 7.71 (dd, J = 8.2, 2.4 Hz, 1H), 7.30 (d, J = 8.2 Hz, 1H), 7.21 (dd, J = 8.7, 7.2 Hz, 2H), 6.93 (d, J = 8.1 Hz, 2H), 6.77 (t, J = 7.3 Hz, 1H), 4.29 (t, J = 6.2 Hz, 2H), 3.29 (s, 2H), 3.13 (s, 4H), 2.64 (s, 3H), 2.55 (s, 4H), 2.31 (s, 3H), 2.28 (s, 3H), 1.95 (p, 2H). Nuclear magnetic resonance carbon spectrum: 13C NMR (176MHz, DMSO-d6) δ 158.23, 153.79, 151.54, 149.26, 137.77, 136.37, 136.21, 136.04, 130.48, 129.41, 123.36, 119.82, 119.27, 115.82, 107.38, 74.88, 54.21, 53.11, 48.68, 25.42, 20.82, 20.10, 19.48. MS (ESI) m / z 412.30 ([M+H]+: 412.23).

[0121] Example 22:

[0122] This example provides 1-(3,4-dimethylphenyl)-6-methyl-5-{[4-(1,4-oxazepan-4- yl)butyl]oxy}-4,5-dihydropyr azolo [3,4-d] py rimidin-4-one, which is synthesized according to the procedure of Example 19, except that 1,3-dibromopropane is replaced by 1,4-dibromobutane and piperidine is replaced by morpholine, to give the second stage intermediate compound as 5-[(4-bromobutyl)oxy]-1-(3,4-dimethylphenyl)-6-methyl-4,5-dihydropyr azolo [3,4-d] py rimidin-4-one.

[0123] Nuclear magnetic resonance hydrogen spectrum: 1H NMR (600 MHz, DMSO-d6) δ 8.31 (s, 1H), 7.74 (s, 1H), 7.73-7.66 (m, 1H), 7.31 (d, J = 8.1 Hz, 1H), 4.24 (t, J = 6.5 Hz, 2H), 3.57 (s, 4H), 3.30 (s, 2H), 2.60 (s, 3H), 2.34 (t, 4H), 2.31 (s, 3H), 2.28 (s, 3H), 1.76 (p, J = 6.7 Hz, 2H), 1.62 (p, 2H). Nuclear magnetic resonance carbon spectrum: 13C NMR (176 MHz, DMSO-d6) δ 158.18, 153.78, 149.25, 137.79, 136.38, 136.21, 136.07, 130.49, 123.38, 119.84, 107.38, 76.60, 66.74, 58.18, 53.83, 25.92, 22.69, 20.95, 20.11, 19.49. MS (ESI) m / z 487.34 ([M+H]+: 487.28).

[0124] Example 23:

[0125] This example provides 1-(3,4-dimethylphenyl)-6-methyl-5-{[4-(4- phenylpiperazin-1-yl)butyl]oxy}-4,5-dihydropyrrolo[3,4-d]pyrimidin-4-one, which is synthesized according to the procedure of Example 19, except that 1,3-dibromopropane is replaced by 1,4-dibromobutane and piperidine is replaced by 1-phenylpiperazine, to give the second stage intermediate compound as 5-[(4-bromobutyl)oxy]-1-(3,4-dimethylphenyl)-6-methyl-4,5-dihydropyrrolo[3,4-d]pyrimidin-4-one.

[0126] Nuclear magnetic resonance hydrogen spectrum: 1H NMR (600 MHz, DMSO-d6) δ 8.31 (s, 1H), 7.74 (d, J = 2.3 Hz, 1H), 7.71 (dd, J = 8.1, 2.4 Hz, 1H), 7.31 (d, J = 8.1 Hz, 1H), 7.20 (dd, J = 8.8, 7.2 Hz, 2H), 6.93 (d, J = 8.2 Hz, 2H), 6.77 (t, J = 7.2 Hz, 1H), 4.26 (t, J = 6.4 Hz, 2H), 3.29 (s, 2H), 3.13 (s, 4H), 2.61 (s, 3H), 2.57 - 2.51 (m, 4H), 2.31 (s, 3H), 2.28 (s, 3H), 1.79 (p, J = 6.7 Hz, 2H), 1.67 (p, 2H). Nuclear magnetic resonance carbon spectrum: 13C NMR (176 MHz, DMSO-d6) δ 158.17, 153.78, 151.56, 149.24, 137.78, 136.38, 136.21, 136.05, 130.49, 129.41 123.37, 119.82, 119.25, 115.83, 107.39, 76.61, 57.75, 53.23, 48.71, 25.98, 23.02, 20.96, 20.11, 19.49. MS (ESI) m / z 487.34 ([M+H]+: 487.28).

[0127] Example 24:

[0128] This example provides 1 -(4-fluorophenyl)-5-(3-(4-fluoropiperidin-1 -yl)propyl)-6- methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, the synthesis of which is the same as Example 16, except that piperidine is replaced by 4-fluoropiperidine.

[0129] Nuclear magnetic resonance hydrogen spectrum: 1H NMR (700 MHz, DMSO-d6) δ 8.33 (s, 1H), δ 8.33 (S, 1H), δ 8.03 (m, 2H), δ 7.40 (q, 2H, J = 21 Hz), δ 4.25 (t, 2H, J = 7 Hz), δ 2.98 (d, 2H, J = 7 Hz), δ 2.61 (S, 3H), δ 2.61 (S, 3H), δ 2.40 (S, 1H), δ 1.98 (t, 2H, J = 28 Hz), δ 1.91 (p, 2H, 28 Hz), δ 1.78 (d, 2H, 7 Hz), δ 1.44 (m, 2H).

[0130] Example 25:

[0131] This example provides 5-(3-(4,4-difluoropiperidin-l-yl)propyl)-l-(4- fluorophenyl)-6-methyl-l,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, which is synthesized according to the procedure of Example 16, except that piperidine is replaced by 4,4-difluoropiperidine.

[0132] Nuclear magnetic resonance hydrogen spectrum: 1H NMR (700 MHz, DMSO) δ 8.33 (t, 1H, J = 14 Hz), δ 8.01 (p, 2H, J = 21 Hz), δ 7.41 (m, 2H), δ 4,67 (d, 1H, J = 49 Hz), δ 4.24 (t, 2H, J = 14 Hz), δ 2.61 (d, 3H, J = 7 Hz), δ 2.48 (t, 2H, J = 14 Hz), δ 1.90 (q, 4H, J = 28 Hz), δ 1.70 (m, 4H), δ 1.22 (s, 2H).

[0133] Example 26:

[0134] This example provides 1-(4-fluorophenyl)-6-methyl-5-(3-(4-(trifluoromethyl)piperidin-l- yl)propyl)-l,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, which is synthesized according to the procedure of Example 16, except that piperidine is replaced by 4-(trifluoromethyl)piperidine.

[0135] Nuclear magnetic resonance hydrogen spectrum: 1H NMR (700 MHz, DMSO) δ 8.33 (t, 1H, J = 14 Hz), δ 8.01 (p, 2H, J = 21 Hz), δ 7.41 (m, 2H), δ 4,67 (d, 1H, J = 49 Hz), δ 4.24 (t, 2H, J = 14 Hz), δ 2.61 (d, 3H, J = 7 Hz), δ 2.48 (t, 2H, J = 14 Hz), δ 1.90 (q, 4H, J = 28 Hz), δ 1.70 (m, 4H), δ 1.22 (s, 2H).

[0136] The effectiveness of the compounds of the present application is demonstrated below from the pharmacological aspect.

[0137] 1. Preparation of σ1 receptor membrane and determination of ligand affinity

[0138] Preparation of σ1 receptor membrane: guinea pig decapitation, operation on ice, quickly take the brain, put the tissue into a centrifuge tube, add 0.01M Tris HCl + 0.32M sucrose solution, homogenize at 4 for 3-4s, homogenize 4 times, then add 0.01M Tris HCl + 0.32M sucrose solution, adjust to 10ml / g, use a balance to adjust the weight of the homogenized test tube, centrifuge at 1000r for 10min; take the supernatant and add 0.01M Tris HCl + 0.32M sucrose solution to adjust to 2ml / g, centrifuge at 1000r, 4℃ for 10min; take the supernatant and centrifuge at 11500r, 4℃ for 25min; take the precipitate and add 0.01M Tris HCl + 0.32M sucrose solution to adjust to 3mL / g, incubate at 25℃ for 15min, centrifuge at 11500r, 4℃ for 25min, store the precipitate at -80℃ for standby.

[0139] Receptor competitive binding experiment:

[0140] Experimental materials and instruments:

[0141] Flupentixol was purchased from Sigma-Aldrich company; PPO, POPOP and lipid-soluble scintillation solution were purchased from Shanghai Reagent Factory; radioactive ligand [3H]-(+)-pentazocine; Wallace 1450 MicroBeta TriLux scintillation luminescence counter, Perkin Elmer product.

[0142] Experimental method:

[0143] (1) First, the prepared membrane is dispersed uniformly with an appropriate amount of homogenate liquid, and an appropriate amount of homogenate liquid is added to prepare a 50mL membrane suspension, which is ready for use; (2) 100μL of membrane preparation is added to each reaction tube; (3) 100μL of B liquid is added to the total binding tube (TB), 100μL of flupentixol (final concentration 10-5M) is added to the non-specific binding tube (NB), and 100μL of the test compound (final concentration 10-5M) is added to each specific binding tube (SB) of the test compound; (4) 10μL of radioactive ligand [3H]-(+)-pentazocine (final concentration 4nM) is added to each reaction tube; (5) each reaction tube is incubated at 25℃ for 3h, after the reaction is completed, the bound ligand is quickly filtered under reduced pressure, Whatman filter paper is saturated with 0.25% PEI solution 2h in advance, and the filter paper is washed with ice-cold test buffer, then the filter paper is taken out and placed in a 2ml scintillation vial, 1mL of toluene scintillation liquid is added and mixed well; (6) the scintillation vial is placed in a liquid scintillation counter for counting;

[0144] Data processing:

[0145] TB: total and constant;

[0146] NB: non-specific binding constant;

[0147] SB: binding constant of the compound;

[0148] Inhibition rate (I%) = (TB - SB) / (TB - NB) x 100%;

[0149] logit method to calculate the IC of each compound 50 ;

[0150] Scatchard plot to obtain the Ka value and Bmax of each radioligand;

[0151] Finally, the Ki value of the measured compound is obtained, Ki = IC 50 / (1 + C / Ka).

[0152] Table 1 Ki values of compounds in Examples 1-26

[0153]

[0154]

[0155]

[0156]

[0157]

[0158] 2. Formaldehyde-induced mouse pain model experiment

[0159] Experimental animals: healthy ICR mice, male, 22-40 g, provided by Nanjing Qinglongshan Animal Breeding Center;

[0160] Main reagents: positive drug (Pregabalin); formaldehyde solution, purchased from Longxi Chemical; sodium chloride injection, purchased from Xuzhou Fifth Pharmaceutical Factory Co., Ltd.; compound prepared in Example 16;

[0161] Experimental method: the mice were randomly divided into negative control group, model group, positive drug group (pregabalin 40 mg / kg) and the compound prepared in Example 16 high, medium and low dose groups (specific dosing 20 mg / kg, 10 mg / kg, 5 mg / kg), 10 in each group; the negative control group and the model group were given the corresponding solvent double distilled water by gavage, the positive drug group was given the corresponding positive drug by gavage, and each dose group of the compound was given the corresponding dose of the compound by gavage, and the gavage volume was 0.1 mL / 10 g; the model group, the positive drug group and each dose group of the compound were injected 20 μL of 2.5% formaldehyde solution subcutaneously in the left hind foot of the mice 15 min after gavage to form a skin bump as the success standard of modeling, and the negative control group of mice was injected 20 μL of normal saline subcutaneously in the left hind foot; after the modeling was successful, the time of the mice licking and biting the injection site of the foot was observed at 0-5 min and 15-45 min;

[0162] Experimental results: the time statistics results of the mice licking and biting the injection site of the foot at 0-5 min and 15-45 min are shown in Figure 1 、 Figure 2 It can be seen from Figure 1 、 Figure 2 that the compound of the application can significantly improve the I phase and II phase pain induced by formalin.

[0163] Example 27:

[0164] A tablet, the formula of which is: active ingredient (any one of the target products in Examples 1-26 or a pharmaceutically acceptable salt thereof) 100 mg, microcrystalline cellulose 50 mg, lactose 50 mg, povidone K30 9 mg, sodium carboxymethyl starch 12 mg, silicon dioxide 2.5 mg, and magnesium stearate 1.5 mg;

[0165] The raw and auxiliary materials were sieved through an 80-mesh sieve, the prescription amount of active ingredient, microcrystalline cellulose, lactose and povidone K30 were weighed, added to a high-speed mixing granulator, and stirred and mixed uniformly at low speed, then an appropriate amount of purified water was added and stirred at low speed, and then high-speed cutting granulation was performed, and the wet granules were dried at 60°C for 3 h, and then the granules were sized through a 24-mesh sieve, and then the prescription amount of sodium carboxymethyl starch, silicon dioxide and magnesium stearate were added, and then total mixing was performed, and then a rotary tablet press was used to press the tablets.

Claims

1. A pyrazolo[3,4-d]pyrimidin-4-one derivative, characterized in that: The general structural formula of the pyrazolo[3,4-d]pyrimidin-4-one derivatives is as follows: ； In the above formula, represents a group selected from hydrogen, halogen or alkyl; m is an integer from 3 to 5; n is an integer from 0 to 2; X is a carbon atom, an oxygen atom or a nitrogen atom; represents a group selected from hydrogen, phenyl, and alkyl; the alkyl group is methyl.

2. A pyrazolo[3,4-d]pyrimidin-4-one derivative according to claim 1, characterized in that: The halogen is fluorine.

3. A pyrazolo[3,4-d]pyrimidin-4-one derivative, characterized in that: The pyrazolo[3,4-d]pyrimidin-4-one derivative is selected from any one of the following compounds: 5-{[3-(Hexahydropyran-1-yl)propyl]oxy}-6-methyl-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one; 6-methyl-5-{[3-(1,4-oxazinan-4-yl)propyl]oxy}-1-phenyl-4,5-dihydro-pyrazolo[3,4-d]pyrimidin-4-one; 6-methyl-5-{[3-(4-methylpiperazin-1-yl)propyl]oxy}-1-phenyl-4,5-dihydro-1H-pyrazolo[3,4-d]pyrimidin-4-one; 6-methyl-1-phenyl-5-{[3-(4-phenylpiperazin-1-yl)propyl]oxy}-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one; 6-methyl-1-phenyl-5-{[3-(tetrahydro-1H-pyrrol-1-yl)propyl]oxy}-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one; 5-{[3-(Azolidin-1-yl)propyl]oxy}-6-methyl-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one; 6-methyl-5-{[2-(1,4-oxazepan-4-yl)ethyl]oxy}-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one; 6-methyl-5-{[4-(1,4-oxazepan-4-yl)butyl]oxy}-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one; 6-methyl-5-{[4-(4-methylpiperazin-1-yl)butyl]oxy}-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one; 6-methyl-1-phenyl-5-{[4-(4-phenylpiperazin-1-yl)butyl]oxy}-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one; 5-{[5-(Hexahydropyridin-1-yl)pentyl]oxy}-6-methyl-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one; 6-methyl-5-{[5-(1,4-oxazinan-4-yl)pentyl]oxy}-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one; 6-methyl-5-{[5-(4-methylpiperazin-1-yl)pentyl]oxy}-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one; 6-methyl-1-phenyl-5-{[5-(4-phenylpiperazin-1-yl)pentyl]oxy}-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one; 6-methyl-1-phenyl-5-{[5-(tetrahydro-1H-pyrrol-1-yl)pentyl]oxy}-4,5-dihydro-1H-pyrazolo[3,4-d]pyrimidin-4-one; 1-(4-Fluorophenyl)-5-{[3-(hexahydropyridin-1-yl)propyl]oxy}-6-methyl-4,5-dihydro-1H-pyrazolo[3,4-d]pyrimidin-4-one; 1-(4-Fluorophenyl)-6-methyl-5-{[3-(1,4-oxazinan-4-yl)propyl]oxy}-4,5-dihydro-1H-pyrazolo[3,4-d]pyrimidin-4-one; 1-(4-Fluorophenyl)-6-methyl-5-{[4-(1,4-oxazinan-4-yl)butyl]oxy}-4,5-dihydro-1H-pyrazolo[3,4-d]pyrimidin-4-one; 1-(3,4-dimethylphenyl)-5-{[3-(hexahydropyridin-1-yl)propyl]oxy}-6-methyl-4,5-dihydro-1H-pyrazolo[3,4-d]pyrimidin-4-one; 1-(3,4-dimethylphenyl)-6-methyl-5-{[3-(1,4-oxazepan-4-yl)propyl]oxy}-4,5-dihydro-1H-pyrazolo[3,4-d]pyrimidin-4-one; 1-(3,4-dimethylphenyl)-6-methyl-5-{[3-(4-phenylpiperazin-1-yl)propyl]oxy}-4,5-dihydro-1H-pyrazolo[3,4-d]pyrimidin-4-one; 1-(3,4-dimethylphenyl)-6-methyl-5-{[4-(1,4-oxazepan-4-yl)butyl]oxy}-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one; 1-(3,4-dimethylphenyl)-6-methyl-5-{[4-(4-phenylpiperazin-1-yl)butyl]oxy}-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one; 1-(4-Fluorophenyl)-5-(3-(4-fluoropiperidin-1-yl)propyl)-6-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one; 5-(3-(4,4-difluoropiperidin-1-yl)propyl)-1-(4-fluorophenyl)-6-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one; 1-(4-Fluorophenyl)-6-methyl-5-(3-(4-(trifluoromethyl)piperidin-1-yl)propyl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one.

4. A pharmaceutical composition, characterized in that: The invention comprises a therapeutically effective amount of the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

5. Use of the pharmaceutical composition according to claim 4 in the preparation of a medicament for preventing or treating pain diseases.

6. The use according to claim 5, characterized in that: The pain disease is neuralgia.

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