Pyrazolo [3, 4-d] pyrimidine-4-ketone derivative and application thereof

Abstract

The invention belongs to the technical field of medicinal chemistry, and discloses application of pyrazolo [3, 4-d] pyrimidine-4-ketone derivatives with the following structure or pharmaceutically acceptable salts thereof in prevention or treatment of pain diseases, # imgabs0 # R1 represents a substituent 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 substituent selected from hydrogen, phenyl, and substituted or unsubstituted alkyl.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical chemistry, and specifically relates to a pyrazolo[3,4-d]pyrimidine-4-one derivative and an application thereof in treating pain diseases. Background Art

[0002] Pain is one of the most common clinical symptoms. According to 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 abnormal due to trauma or disease, spontaneous impulses will be generated, causing pain at the origin of the nerve. This is called neuropathic pain (NP), which is usually referred to as neuralgia. However, existing neuralgia analgesics still have limited analgesic efficiency and low response rate in clinical use. There are also side effects and adverse reactions of varying degrees. In particular, pregabalin may increase respiratory depression or even death when used in combination with high-dose opioids.

[0003] σ 1 The receptors are widely expressed in multiple areas of the nervous system, and are involved in regulating neuropathic pain and regulating the activity of neuroimmune cells including microglia, thereby affecting the inflammatory response process. 1 The receptor works by directly interacting with the opioid μ receptor. 1 In the formalin-induced pain experiment conducted on mice with receptor gene knockout, there was no obvious pain response in phase I and phase II. The same phenomenon occurred in the capsaicin-induced neuropathic hyperalgesia model. 1 Mice with knockout of the receptor gene also showed insensitive behavior in the paclitaxel-induced cold hyperalgesia and mechanical hyperalgesia models and the sciatic nerve ligation neuralgia model. 1 Receptor antagonists are good anti-inflammatory and analgesic targets. 1 Receptor antagonists themselves have analgesic effects on pain, especially neuralgia. 1 There is great interest in receptor antagonists. Currently, S1RA is used in experiments to treat various pains and has entered the clinical stage. Among them, experiments to treat neuropathic pain alone have entered clinical research. 1 The receptor antagonists showed similar effects on pain model in normal mice. 1 The above experiments prove that there is a similar phenomenon of pain desensitization in mice with receptor knockout. 1Haloperidol and its metabolites, which have sigma receptor antagonist effects, have significant analgesic effects in various neuropathic pain models. 1 Receptor antagonists are used in anti-pain treatment and have important scientific value and social significance in the clinical treatment of pain and neuralgia. Summary of the invention

[0004] The purpose of the present invention is to provide a pyrazolo[3,4-d]pyrimidine-4-one derivative and its application in treating pain diseases in view of the above problems existing in the prior art.

[0005] To achieve the above objectives, the technical solution of the present invention is as follows:

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

[0007]

[0008] In the above formula, R 1 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; R 2 represents a group selected from hydrogen, phenyl, and substituted or unsubstituted alkyl.

[0009] The alkyl group 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-(Hexahydropyran-1-yl)propyl]oxy}-6-methyl-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one;

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

[0029] 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;

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

[0032] 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;

[0033] 1-(3,4-dimethylphenyl)-6-methyl-5-{[4-(1,4-oxazepan-4-yl)butyl]oxy}-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one;

[0034] 1-(3,4-dimethylphenyl)-6-methyl-5-{[4-(4-phenylpiperazin-1-yl)butyl]oxy}-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one;

[0035] 1-(4-Fluorophenyl)-5-(3-(4-fluoropiperidin-1-yl)propyl)-6-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0036] 5-(3-(4,4-difluoropiperidin-1-yl)propyl)-1-(4-fluorophenyl)-6-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0037] 1-(4-Fluorophenyl)-6-methyl-5-(3-(4-(trifluoromethyl)piperidin-1-yl)propyl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one.

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

[0039] The salts are salts containing pharmaceutically acceptable anions, including: hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate or bisulfate, phosphate or acid phosphate, acetate, lactate, citrate, tartrate, maleate, fumarate, methanesulfonate, gluconate, saccharate, benzoate, ethanesulfonate, benzenesulfonate, and p-toluenesulfonate.

[0040] The pharmaceutically acceptable excipients include carriers, excipients, disintegrants, sweeteners, lubricants, etc.; the carriers include inert solid fillers, diluents, sterile aqueous solutions or organic solutions; for the convenience of oral treatment, the target product or its pharmaceutically acceptable salt is prepared together with excipients in the form of 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. Those skilled in the art can determine appropriate dosage based on these or other factors. The effective dose of central nervous system drugs commonly used is well known to the technician, and the total daily dose is usually between 0.05mg and 2000mg.

[0042] The pharmaceutical composition can be administered by any appropriate route, for example, orally in capsule form, parenterally in the form of an injection, topically in the form of an ointment or lotion, rectally in the form of a suppository, or transdermally in the form of a patch delivery system.

[0043] In a third aspect, the present invention provides use of the aforementioned pharmaceutical composition in preventing or treating pain diseases.

[0044] The pain diseases refer to acute pain (such as pain caused by acute injury of soft tissue and joints, postoperative pain, obstetric pain, acute herpes zoster pain, gout, etc.), chronic pain (such as pain caused by strain or degeneration of soft tissue and joints, disc-related pain, neuropathic pain, etc.), intractable pain (such as trigeminal neuralgia, postherpetic neuralgia, herniated disc, intractable headache, etc.), cancer pain (such as pain caused by advanced tumors, pain caused by tumor metastasis, etc.), and special pain (such as thromboangiitis vasculitis, intractable angina pectoris, idiopathic chest and abdominal pain, etc.).

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

[0046] The present invention provides a pyrazolo[3,4-d]pyrimidine-4-one derivative, and proves that the compound has a σ 1 The compound has a selective antagonist effect on the receptors. Animal test results have shown that the compound can significantly improve formalin-induced phase I and phase II pain. 1 The compounds of the present invention are closely related to the reactions regulated by the receptor-mediated nervous system, especially pain. Therefore, the compounds of the present invention have the effect of treating pain, especially neuralgia, and provide a new path for the preparation of drugs for preventing or treating pain diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 These are the results of formalin-induced phase I pain experiment in mice.

[0048] Figure 2 These are the results of formalin-induced phase II pain experiment in mice. DETAILED DESCRIPTION

[0049] The present invention is further described in detail below in conjunction with specific implementations and drawings.

[0050] The general synthetic route of the pyrazolo[3,4-d]pyrimidin-4-one derivatives of the present invention is: firstly synthesize the pyrazolo[3,4-d]pyrimidin-4-one mother body, and then link it with a nitrogen-containing structure through a carbon chain.

[0051] Embodiment 1:

[0052] This embodiment provides 5-{[3-(hexahydropyran-1-yl)propyl]oxy}-6-methyl-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one, and the synthesis steps are as follows:

[0053] The first stage, such as Figure 1As shown, (2E)-2-cyano-3-ethoxyprop-2-enoic acid ethyl ester and phenylethazine are dissolved in anhydrous ethanol solution, and refluxed and reacted for 5 hours to obtain 5-amino-1-phenylpyrazole-4-carboxylic acid ethyl ester, 5-amino-1-phenylpyrazole-4-carboxylic acid ethyl ester and sodium hydroxide are mixed in methanol, condensed and refluxed, and heated to react for 4 hours, and hydrolyzed to obtain 5-amino-1-phenylpyrazole-4-carboxylic acid, and then 5-amino-1-phenylpyrazole-4-carboxylic acid is mixed with acetic anhydride, condensed and refluxed, and heated to react for 4 hours. Filter, dry and crystallize to obtain 6-methyl-1-phenyl-4H-pyrazolo[3,4-d][1,3]oxazacyclohexan-4-one. Finally, mix 6-methyl-1-phenyl-4H-pyrazolo[3,4-d][1,3]oxazacyclohexan-4-one with hydroxylamine hydrochloride in dry pyridine, heat and 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] In the second stage, referring to the Williamson ether synthesis method, in an acetone solution, with anhydrous potassium carbonate as a catalyst, the intermediate compound of the first stage is mixed with 1,3-dibromopropane, and heated in an oil bath at 60°C for 5-8 hours. The molar ratio of the feed materials is about 1:2 to obtain the intermediate compound of the first stage 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] In the third stage, the intermediate compound in the second stage is mixed with piperidine to obtain a reaction product; the reaction formula of the third stage is as follows:

[0058]

[0059] H NMR: 1H NMR (600 MHz, DMSO-d6) δ8.37 (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 (176MHz, 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] Embodiment 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 steps are the same as those in Example 1, except that piperidine is replaced by morpholine.

[0062] H NMR: 1H NMR (600 MHz, DMSO-d6) δ8.36 (s, 1H), 8.02 (dd, J = 8.7, 1.2 Hz, 2H), 7.57 (dd, J = 8.6, 7.3 Hz, 2H), 7.42 (t, J = 7.4 Hz, 1H), 4.29 (d, J = 5.7 Hz, 2H), 3.58 (s, 4H), 3.29 (s, 2H), 2.64 (s, 3H), 2.38 (p, J = 1.9 Hz, 4H), 1.92 (s, 2H). Nuclear magnetic resonance carbon spectrum: 13C NMR (176MHz, DMSO-d6) δ 158.44, 153.78, 149.47, 138.56, 136.61, 129.78, 127.73, 122.25, 107.61, 74.83, 66.67, 54.59, 53.65, 25.09, 20.80. MS (ESI) m / z 370.24 ([M+H]+).

[0063] Embodiment 3:

[0064] This embodiment 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 steps are the same as those in Example 1, except that piperidine is replaced by 1-methylpiperazine.

[0065] H NMR: 1H NMR (600 MHz, DMSO-d6) δ8.36 (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). Carbon nuclear magnetic resonance 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] Embodiment 4:

[0067] This embodiment provides 6-methyl-1-phenyl-5-{[3-(4-phenylpiperazin-1-yl)propyl]oxy}-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one. The synthesis steps are the same as those in Example 1, except that piperidine is replaced by 1-phenylpiperazine.

[0068] H NMR: 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 (176MHz 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] Embodiment 5:

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

[0071] H NMR: 1H NMR (600 MHz, DMSO-d6) δ8.38 (s, 1H), 8.02 (dd, J = 8.7, 1.1 Hz, 2H), 7.58 (dd, J = 8.6, 7.4 Hz, 2H), 7.42 (t, J = 7.4 Hz, 1H), 4.32 (t, J = 6.0 Hz, 2H), 3.08 (t, J = 27.3 Hz, 6H), 2.64 (s, 3H), 2.1-2.04 (m, 2H), 1.86 (s, 4H). Nuclear magnetic resonance carbon spectrum: 13C NMR (176MHz, DMSO-d6) δ 158.32, 153.90, 149.46, 138.52, 136.63, 129.81, 127.79, 122.28, 107.56, 74.56, 53.89, 51.88, 23.30, 21.58, 20.85. MS (ESI) m / z 354.24 ([M+H]+: 354.19).

[0072] Embodiment 6:

[0073] This example provides 5-{[3-(azacyclopentane-1-yl)propyl]oxy}-6-methyl-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one. The synthesis steps are the same as those in Example 1, except that piperidine is replaced with cycloheximide.

[0074] H NMR: 1H NMR (700 MHz, DMSO-d6) δ8.37 (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] Embodiment 7:

[0076] This embodiment provides 6-methyl-5-{[2-(1,4-oxazepan-4-yl)ethyl]oxy}-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one. The synthesis steps are the same as those in Example 1, except that the intermediate compound 5-hydroxy-6-methyl-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one in the first stage is reacted with 4-(2-chloroethyl)-1,4-oxazacyclohexane to obtain the target product. The reaction formula is as follows:

[0077]

[0078] H NMR: 1H NMR (700 MHz, DMSO-d6) δ8.36 (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). Carbon nuclear magnetic resonance spectrum: 13C NMR (176MHz, 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] Embodiment 8:

[0080] This embodiment provides 6-methyl-5-{[4-(1,4-oxazepan-4-yl)butyl]oxy}-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one. The synthesis steps are the same as those in Example 1, except that 1,3-dibromopropane is replaced by 1,4-dibromobutane, and piperidine is replaced by morpholine, and the intermediate product of the second stage is 5-[(4-bromobutyl)oxy]-6-methyl-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one.

[0081] H NMR: 1H NMR (600 MHz, DMSO-d6) δ8.37 (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). Carbon nuclear magnetic resonance 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] Embodiment 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 steps are the same as those in Example 8, except that morpholine is replaced by 1-methylpiperazine.

[0084] H NMR: 1H NMR (600 MHz, DMSO-d6) δ8.36 (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). Carbon NMR 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] Embodiment 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 steps are the same as those in Example 8, except that morpholine is replaced by 1-phenylpiperazine.

[0087] H NMR: 1H NMR (600 MHz, DMSO-d6) δ8.36 (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). Carbon nuclear magnetic resonance spectrum: 13C NMR (176MHz, 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] Embodiment 11:

[0089] This embodiment provides 5-{[5-(hexahydropyridin-1-yl)pentyl]oxy}-6-methyl-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one. The synthesis steps are the same as those in Example 1, except that 1,3-dibromopropane is replaced with 1,5-dibromopentane, and the intermediate product of the second stage is 5-[(5-bromopentyl)oxy]-6-methyl-1-phenyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one.

[0090] H NMR: 1H NMR (600 MHz, DMSO-d6) δ8.36 (s, 1H), 8.02 (dd, J = 8.7, 1.1 Hz, 2H), 7.57 (dd, J = 8.6, 7. 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 (176MHz, 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] Embodiment 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 steps are the same as those in Example 11, except that piperidine is replaced by morpholine.

[0093] H NMR: 1H NMR (600 MHz, DMSO-d6) δ8.36 (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). Carbon nuclear magnetic resonance spectrum: 13C NMR (176MHz, 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] Embodiment 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, and its synthesis steps are the same as those of Example 11, except that piperidine is replaced by 1-methylpiperazine.

[0096] H NMR: 1H NMR (600 MHz, DMSO-d6) δ8.36 (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 (176MHz, 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] Embodiment 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, and its synthesis steps are the same as those of Example 11, except that piperidine is replaced by 1-phenylpiperazine.

[0099] H NMR: 1H NMR (600 MHz, DMSO-d6) δ8.36 (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=7.2Hz,1H),4.25(t,J=6.4Hz,2H),3.13(s,4H),2.62(s,3H),2.58-2.5 1(m,4H),2.37(s,2H),1.78(p,J=6.7Hz,2H),1.61-1.53(m,2H),1.53-1.45(m,2H). Nuclear magnetic resonance carbon spectrum: 13C NMR (176MHz, 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] Embodiment 15:

[0101] This example provides 6-methyl-1-phenyl-5-{[5-(tetrahydro-1H-pyrrol-1-yl)pentyl]oxy}-4,5-dihydro-1H-pyrazolo[3,4-d]pyrimidin-4-one, and its synthesis steps are the same as those of Example 11, except that piperidine is replaced by pyrrolidine.

[0102] H NMR: 1H NMR (600 MHz, DMSO-d6) δ8.36 (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). Carbon nuclear magnetic resonance 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] Embodiment 16:

[0104] This embodiment provides 1-(4-fluorophenyl)-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 phenyl ethazane is replaced with (4-fluorophenyl) ethazane, (2E)-2-cyano-3-ethoxyprop-2-enoic acid ethyl ester (1) and (4-fluorophenyl) ethazane are dissolved in anhydrous ethanol solution and reacted by reflux heating for 5 hours to obtain 5-amino-1-(4-fluorophenyl)pyrazole-4-carboxylic acid ethyl ester, 5-amino-1-(4-fluorophenyl)pyrazole-4-carboxylic acid ethyl ester is mixed with sodium hydroxide in methanol, and condensed by reflux heating to hydrolyze to obtain 5-amino-1-(4-fluorophenyl)pyrazole-4-carboxylic acid, and then 5-amino-1-(4-fluorophenyl)pyrazole-4-carboxylic acid ethyl ester is reacted by reflux heating. The mixture of 1-(4-fluorophenyl)-6-methyl-4H-pyrazolo[3,4-d][1,3]oxazahexacyclohexan-4-one and hydroxylamine hydrochloride was mixed in dry pyridine, and the mixture was heated under reflux for 8 hours to obtain the intermediate compound of the first stage, namely, 1-(4-fluorophenyl)-5-hydroxy-6-methyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one; and the intermediate compound of the second stage, namely, 5-[(3-bromopropyl)oxy]-1-(4-fluorophenyl)-6-methyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one.

[0105] H NMR: 1H NMR (600 MHz, DMSO-d6) δ8.36 (s, 1H), 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 (176MHz, 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] Embodiment 17:

[0107] This example provides 1-(4-fluorophenyl)-6-methyl-5-{[3-(1,4-oxazin-4-yl)propyl]oxy}-4,5-dihydro-1H-pyrazolo[3,4-d]pyrimidin-4-one. Its synthesis steps are the same as those in Example 16, except that piperidine is replaced by morpholine.

[0108] H NMR: 1H NMR (600 MHz, DMSO-d6) δ8.36 (s, 1H), 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 (176MHz, 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] Embodiment 18:

[0110] This embodiment provides 1-(4-fluorophenyl)-6-methyl-5-{[4-(1,4-oxazin-4-yl)butyl]oxy}-4,5-dihydro-1H-pyrazolo[3,4-d]pyrimidin-4-one. The synthesis steps are the same as those in Example 16, except that 1,3-dibromopropane is replaced by 1,4-dibromobutane, and piperidine is replaced by morpholine, to obtain the second-stage intermediate compound 5-[(4-bromobutyl)oxy]-1-(4-fluorophenyl)-6-methyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one.

[0111] H NMR: 1H NMR (600 MHz, DMSO-d6) δ8.37 (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 (176MHz, 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] Embodiment 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)ethazine, 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 condensed and refluxed to obtain 5-amino-1-(3,4-dimethylphenyl) pyrazole-4-carboxylic acid. Then 5-amino-1-(3,4-dimethylphenyl) pyrazole-4-carboxylic acid ethyl ester is hydrolyzed. (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]oxazacyclohexan-4-one. Finally, 1-(3,4-dimethylphenyl)-6-methyl-4H-pyrazolo[3,4-d][1,3]oxazacyclohexan-4-one was reacted with hydroxyl hydrochloride. The amine is mixed in dry pyridine, heated to reflux for reaction for 8 hours, and the intermediate compound of the first stage 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] H NMR: 1H NMR (600 MHz, DMSO-d6) δ8.31 (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). Carbon nuclear magnetic resonance spectrum: 13C NMR (176MHz, 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] Embodiment 20:

[0116] This embodiment provides 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. Its synthesis steps are the same as those of Example 19, except that piperidine is replaced by morpholine.

[0117] H NMR: 1H NMR (600 MHz, DMSO-d6) δ8.31 (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 (176MHz, 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] Embodiment 21:

[0119] This embodiment provides 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. Its synthesis steps are the same as those of Example 19, except that piperidine is replaced by 1-phenylpiperazine.

[0120] H NMR: 1H NMR (600 MHz DMSO-d6) δ8.31 (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] Embodiment 22:

[0122] This embodiment provides 1-(3,4-dimethylphenyl)-6-methyl-5-{[4-(1,4-oxazepan-4-yl)butyl]oxy}-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one. The synthesis steps are the same as those in Example 19, except that 1,3-dibromopropane is replaced by 1,4-dibromobutane, and piperidine is replaced by morpholine, and the intermediate compound of the second stage is 5-[(4-bromobutyl)oxy]-1-(3,4-dimethylphenyl)-6-methyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one.

[0123] H NMR: 1H NMR (600 MHz, DMSO-d6) δ8.31 (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 (176MHz, 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] Embodiment 23:

[0125] This embodiment provides 1-(3,4-dimethylphenyl)-6-methyl-5-{[4-(4-phenylpiperazin-1-yl)butyl]oxy}-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one. The synthesis steps are the same as those in Example 19, except that 1,3-dibromopropane is replaced by 1,4-dibromobutane, and piperidine is replaced by 1-phenylpiperazine, and the intermediate compound of the second stage is 5-[(4-bromobutyl)oxy]-1-(3,4-dimethylphenyl)-6-methyl-4,5-dihydropyrazolo[3,4-d]pyrimidin-4-one.

[0126] H NMR: 1H NMR(600MHz,DMSO-d6)δ8.31(s,1H),7.74(d,J=2.3Hz,1H),7.71(dd,J=8.1,2.4H z,1H),7.31(d,J=8.1Hz,1H),7.20(dd,J=8.8,7.2Hz,2H),6.93(d,J=8.2Hz,2H),6 .77(t,J=7.2Hz,1H),4.26(t,J=6.4Hz,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=6.7Hz,2H),1.67(p,2H). Nuclear magnetic resonance carbon spectrum: 13C NMR (176MHz, 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] Embodiment 24:

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

[0129] H NMR: 1H NMR (700 MHz, DMSO-d6) δ8.33 (s, 1H), δ8.33 (s, 1H), δ8.03 (m, 2H), δ7.40 (q, 2H, J=21Hz), δ4.25 (t, 2H, J=7Hz), δ2.98 (d, 2H, J=7Hz), δ2.61 (S, 3H), δ2.61 (S, 3H), δ2.40 (S, 1H), δ1.98 (t, 2H, J=28Hz), δ1.91 (p, 2H, 28Hz), δ1.78 (d, 2H, 7Hz), δ1.44 (m, 2H).

[0130] Embodiment 25:

[0131] This embodiment provides 5-(3-(4,4-difluoropiperidin-1-yl)propyl)-1-(4-fluorophenyl)-6-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one. Its synthesis steps are the same as those of Example 16, except that piperidine is replaced by 4,4-difluoropiperidine.

[0132] H NMR: 1H NMR (700 MHz, DMSO-d6) δ8.33 (s, 1H), δ8.01 (q, J=14 Hz, 2H), 7.40 (t, 2H, J=21 Hz), δ4.25 (t, 3H, J=14 Hz), δ2.61 (s, 3H), δ2.51 (s, 2H), δ1.95 (m, 4H), δ1.89 (m, 4H), δ1.21 (s, 2H).

[0133] Embodiment 26:

[0134] This embodiment provides 1-(4-fluorophenyl)-6-methyl-5-(3-(4-(trifluoromethyl)piperidin-1-yl)propyl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one. Its synthesis steps are the same as those of Example 16, except that piperidine is replaced by 4-(trifluoromethyl)piperidine.

[0135] H NMR: 1H NMR (700 MHz, DMSO) δ8.33 (t, 1H, J=14 Hz), δ8.01 (p, 2H, J=21 Hz), δ7.41 (m, 2H), δ4,67 (d, 1H, J=49 Hz), δ4.24 (t, 2H, J=14 Hz), δ2.61 (d, 3H, J=7 Hz), δ2.48 (t, 2H, J=14 Hz), δ1.90 (q, 4H, J=28 Hz), δ1.70 (m, 4H), δ1.22 (s, 2H).

[0136] The effectiveness of the compounds of the present invention is verified from the pharmacological aspect.

[0137] 1. σ 1 Preparation of receptor membranes and determination of ligand affinity

[0138] σ 1Preparation of receptor membrane: guinea pigs were decapitated and operated on ice. The brains were quickly removed and the tissues were put into a centrifuge tube. 0.01M Tris HCl + 0.32M sucrose solution was added and homogenized at gear 4 for 3-4s. Homogenize four times, then add 0.01M Tris HCl + 0.32M sucrose solution and adjust to 10ml / g. The weight of the homogenized test tube was adjusted with a balance and centrifuged at 1000r for 10min. The upper layer was taken and adjusted to 2ml / g with 0.01M Tris HCl + 0.32M sucrose solution. Centrifuge at 1000r and 4℃ for 10min. The supernatant was taken and centrifuged at 11500r and 4℃ for 25min. The precipitate was taken and adjusted to 3mL / g with 0.01M Tris HCl + 0.32M sucrose solution. The precipitate was incubated at 25℃ for 15min and centrifuged at 11500r and 4℃ for 25min. The precipitate was stored at -80℃ for later use.

[0139] Receptor competitive binding assay:

[0140] Experimental materials and instruments:

[0141] Haloperidol was purchased from Sigma-Aldrich; PPO, POPOP and lipid-soluble scintillation fluid were purchased from Shanghai Reagent Factory No. 1; radioligand [3H]-(+)-pentazocine; Wallace 1450MicroBeta TriLux scintillation counter was a product of Perkin Elmer.

[0142] Experimental methods:

[0143] (1) First, the prepared membrane was evenly dispersed with an appropriate amount of homogenate using a homogenizer, and an appropriate amount of homogenate was added to form a 50 mL membrane suspension for later use; (2) 100 μL of the membrane preparation was added to each reaction tube; (3) 100 μL of LB solution was added to the total binding tube (TB), 100 μL of haloperidol (final concentration 10-5 M) was added to the nonspecific binding tube (NB), and 100 μL of the test compound (final concentration 10-5 M) was added to the specific binding tube (SB); (4) The radioligand [3H]-(+)-pentazocine was added to each reaction tube. 10 μL (final concentration 4 nM); (5) Incubate each reaction tube at 25°C for 3 h. After the reaction is complete, the bound ligand is quickly filtered under reduced pressure. The Whatman test paper is saturated with 0.25% PEI solution 2 h in advance and washed thoroughly with ice-cold test buffer. The filter is taken out and placed in a 2 ml scintillation cup. 1 mL of toluene scintillation fluid is added and mixed; (6) The scintillation bottle is placed in a liquid scintillation counter for counting;

[0144] Data processing:

[0145] TB: Summary and constants;

[0146] NB: nonspecific binding constant;

[0147] SB: binding constant of the compound;

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

[0149] Logit method was used to calculate the IC of each compound 50 ;

[0150] The Ka value and Bmax of each radioligand were obtained by Scatchard plotting;

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

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

[0153]

[0154]

[0155]

[0156]

[0157]

[0158] 2. Formalin-induced pain model experiment in mice

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

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

[0161] Experimental method: Mice were randomly divided into negative control group, model group, positive drug group (pregabalin 40 mg / kg) and high, medium and low dose groups (specific dosage 20 mg / kg, 10 mg / kg, 5 mg / kg) of the compound prepared in Example 16, with 10 mice in each group; the negative control group and the model group were gavaged with the corresponding solvent double distilled water, the positive drug group was gavaged with the corresponding positive drug, and the compound dose groups were gavaged with the corresponding dose compound, and the gavage volume was 0.1 mL / 10 g; 15 minutes after gavage, the model group, the positive drug group and the compound dose groups were subcutaneously injected with 20 μL of 2.5% formalin formaldehyde solution in the left hind foot of the mice, and the formation of skin papules was the standard for successful modeling. The negative control group mice were subcutaneously injected with 20 μL of normal saline in the left hind foot; after successful modeling, the time for the mice to lick and bite the injected foot was observed from 0 to 5 minutes and from 15 to 45 minutes;

[0162] Experimental results: The statistical results of the time of mice licking and biting the injected foot at 0-5min and 15-45min are as follows: Figure 1 , Figure 2 shown; from Figure 1 , Figure 2 It can be seen that the compound of the present invention can significantly improve the formalin-induced phase I and phase II pain.

[0163] Embodiment 27:

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

[0165] The raw and auxiliary materials are sieved through an 80-mesh sieve for later use. The prescribed amount of active ingredients, microcrystalline cellulose, lactose, and povidone K30 are weighed and added to a high-speed mixer. Mix evenly at a low speed. An appropriate amount of purified water is added. The mixture is stirred at a low speed and granulated at a high speed. The wet granules are dried at 60°C for 3 h. The granules are sieved through a 24-mesh sieve. The prescribed amount of sodium carboxymethyl starch, silicon dioxide, and magnesium stearate are added. The mixture is mixed and the tablets are compressed using a rotary tablet press.

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, 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; R2 represents a group selected from hydrogen, phenyl, substituted or unsubstituted alkyl.

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

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

4. A pyrazolo[3,4-d]pyrimidin-4-one derivative according to claim 1, 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.

5. A pharmaceutical composition, characterized in that: The invention comprises a therapeutically effective amount of the compound according to claim 1-7 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

6. Use of the pharmaceutical composition according to claim 5 in preventing or treating pain diseases.

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

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