Dihydroquinolinone compound and application thereof

Abstract

The invention relates to the technical field of biological medicine, in particular to a novel skeleton dihydroquinolinone compound and application thereof. The invention provides an 8-substituted dihydroquinolinone compound with a structure as shown in a formula (I) or pharmaceutically acceptable salt or stereoisomer of the 8-substituted dihydroquinolinone compound and application of the 8-substituted dihydroquinolinone compound. The 8-substituted dihydroquinolinone compound disclosed by the invention has remarkable inhibitory activity on phosphodiesterase 5 (PDE5), has a remarkable treatment effect on diseases related to phosphodiesterase 5, such as a pulmonary arterial hypertension animal model, and can be used for preparing a new target candidate drug for pulmonary arterial hypertension.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to an 8-substituted dihydroquinolinone compound and application thereof. Background Art

[0002] Pulmonary artery hypertension (PAH) is a malignant pulmonary vascular disease characterized by progressive pulmonary artery occlusion leading to a gradual increase in pulmonary vascular resistance and subsequent right ventricular failure (RVF). Its pathological features are pulmonary arteriolar intimal proliferation and remodeling. The 5-year survival rate of PAH patients is only about 34%. Therefore, the disease is called "cancer among cardiovascular diseases". Although PAH targeted drug therapy has made significant progress in improving patients' clinical observation indicators (exercise tolerance, hemodynamic parameters, etc.) and delaying the time to clinical deterioration in recent years, the long-term prognosis of patients is still not ideal. The first-line anti-PAH drugs currently used in clinical practice can only slow down the deterioration of the disease and cannot meet the requirements of clinical effective cure. In-depth exploration of the pathogenesis of pulmonary hypertension and efforts to develop new targets that may intervene in pulmonary hypertension are of great significance to new drug development and disease treatment.

[0003] Phosphodiesterases (PDEs) are a superfamily of enzymes in cells that regulate cell signal transduction by catalyzing the degradation of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP). As important second messengers in cells, cyclic nucleotides (cAMP and cGMP) mediate and participate in the occurrence and development of various physiological regulations and diseases in the body. Adenylate cyclase (AC) and guanylate cyclase (GC) synthesize cAMP and cGMP molecules, and degrade cAMP and cGMP through PDEs to maintain a delicate balance of cAMP and cGMP molecular levels in cells. cAMP mainly exerts its functions through protein kinase A (PKA) and cAMP-activated exchange proteins (EPACs), regulating events such as cell movement, migration, hormone secretion, cell metabolism and gene transcription. There are 11 subtypes of PDE in the human body, among which PDE5 can specifically hydrolyze cGMP and is a key regulator of cGMP signal transduction. It is mainly distributed in platelets, vascular smooth muscle, etc. It participates in the NO-cGMP signaling pathway in platelets to regulate cGMP signals in the brain and regulate the contraction of vascular smooth muscle, especially vasoconstriction in the penis and lungs. Currently, PDE5 inhibitors, which are classic marketed drugs for the treatment of PAH, include sildenafil and tadalafil. They inhibit the hydrolysis function of PDE5 to increase the concentration of intracellular cGMP, play a role in vasodilation and inhibit the proliferation of endothelial cells, thereby exerting anti-PAH efficacy. However, existing studies have found that sildenafil has poor pharmacokinetic stability and a short half-life, and long-term use can easily cause muscle pain and visual impairment. Therefore, the discovery of highly active and highly selective PDE5 inhibitors and their important role in the research of new anti-PAH target drugs will bring new opportunities for drug treatment for PAH patients.

[0004] In view of this, the present invention is proposed. Summary of the invention

[0005] In order to solve the above technical problems, the present invention provides a new skeleton dihydroquinolinone compound and its application. The dihydroquinolinone compound has a significant inhibitory effect on phosphodiesterase type 5 and can be used to prepare a new target candidate drug for pulmonary arterial hypertension.

[0006] Specifically, the technical solution of the present invention is as follows: In the first aspect, the present invention provides an 8-substituted dihydroquinolinone compound having a chemical structure as shown in the general formula (I):

[0007] (I)

[0008] In the general formula (I),

[0009] R 1is C3~C6 alkyl or substituted C3~C6 cycloalkyl, heteroaryl or benzobisoxazolyl; R 2 is phenyl or substituted phenyl, C1~C6 alkyl or C3~C6 cycloalkyl; R 3 is C1~C5 alkyl, phenyl or substituted phenyl, pyridyl, or halogenated pyridyl; The substituent of the substituted phenyl group is C1-C5 alkyl, methoxy, or halogen.

[0010] Preferably, R 1 Select from any of the following structures: .

[0011] Preferably, R 2 Select from any of the following structures: .

[0012] Preferably, R 3 Select from any of the following structures: .

[0013] Preferably, the dihydroquinolinone compound has any of the following structures: ; ; ; ; ; ; ; ; ; ; ; .

[0014] The dihydroquinolinone compounds provided by the present invention have significant inhibitory activity against phosphodiesterase 5 (PDE5) type, have significant therapeutic effects on phosphodiesterase 5-related diseases such as pulmonary hypertension animal models, and have no obvious side effects, and can be used to prepare new target candidate drugs for pulmonary hypertension.

[0015] In a second aspect, the present invention provides pharmaceutically acceptable salts or stereoisomers of the aforementioned dihydroquinolinone compounds.

[0016] Based on the structural characteristics of the dihydroquinolinone compounds, their pharmaceutically acceptable salts or stereoisomers also have similar functional effects.

[0017] In a third aspect, the present invention provides a phosphodiesterase 5 activity inhibitor, the main active ingredient of which includes the aforementioned dihydroquinolinone compound or a pharmaceutically acceptable salt or stereoisomer thereof.

[0018] In a fourth aspect, the present invention provides the use of the aforementioned dihydroquinolinone compounds, or pharmaceutically acceptable salts or stereoisomers thereof, or the phosphodiesterase 5 activity inhibitor in the preparation of drugs for treating phosphodiesterase 5 related diseases.

[0019] Preferably, the medicament is for treating pulmonary hypertension.

[0020] Beneficial effects: The present invention provides an 8-substituted dihydroquinolinone compound having a structure as shown in formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof and its application. The dihydroquinolinone compound of the present invention has significant inhibitory activity against phosphodiesterase 5 (PDE5) type, has significant therapeutic effects on phosphodiesterase 5-related diseases such as pulmonary hypertension animal models, and has no obvious side effects, and can be used to prepare new target candidate drugs for pulmonary hypertension. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the drawings required for use in the embodiments or the description of the prior art will be described below.

[0022] Figure 1 The results are those of measuring mean pulmonary artery pressure (mPAP) in rats after administration of compound S-12 (5.0 mg / kg) in Example 14 of the present invention for 21 days.

[0023] Figure 2 This is the result of observing the ventricular wall thickness percentage (WT%) by HE staining 21 days after administration of the compound S-12 (5.0 mg / kg) in Example 14 of the present invention. DETAILED DESCRIPTION

[0024] The technical problem to be solved by the present invention is to provide a new skeleton 8-substituted dihydroquinolinone compound, which has significant PDE5 inhibitory activity in vitro and has good selectivity for other subtypes of PDEs inhibitors, and can reduce the adverse reactions caused by the low selectivity of the compound.

[0025] The compound provided by the present invention has a chemical structure as shown in the general formula (I):

[0026] (I)

[0027] Among them, R 1 is C3~C6 alkyl or substituted C3~C6 cycloalkyl, heteroaryl or benzobisoxazolyl; R 2 is phenyl or substituted phenyl, C1~C6 alkyl or C3~C6 cycloalkyl; R 3 It is C1~C5 alkyl, phenyl or substituted phenyl, pyridyl, halogenated pyridyl; the substituent of the substituted phenyl is C1~C5 alkyl, methoxy, halogen.

[0028] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be described clearly and completely below. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0029] The endpoints and any values ​​of the ranges disclosed in this specification are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of each range, the endpoint values ​​of each range and the individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed in this article.

[0030] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "specific implementations", or "some specific implementations" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the embodiment of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, without contradiction.

[0031] In the examples provided in this specification, if no specific techniques or conditions are specified, the techniques or conditions described in the literature in this field or the product instructions are used. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased through regular channels.

[0032] Example 1

[0033] This example provides compound S-01, whose structural formula and name are as follows: .

[0034] This example provides a method for synthesizing compound S-01, and the specific process is as follows: .

[0035] in: Synthesis of compound L3a: Add L2a (985 mg, 5 mmol) to a 100 ml reaction bottle, dissolve it with 20 mL THF, and add 15 mL p-tolylmagnesium bromide dropwise to the system with a constant pressure funnel under an ice bath. After the addition is complete, reflux for 8 hours, and monitor the reaction by thin layer chromatography (TLC). After the reaction is complete, slowly add 6 ml of 4 mol / L hydrochloric acid to the above reaction system and react overnight. After the reaction is complete by TLC detection, remove the solvent under reduced pressure, adjust the pH of the solution to neutral with a saturated sodium bicarbonate solution, extract twice with dichloromethane, collect the organic layer, dry it over anhydrous sodium sulfate, remove the solvent under reduced pressure, dry it, and obtain 1.05 g of a yellow oil by column chromatography (petroleum ether: ethyl acetate = 20:1), with a yield of 72.4%.

[0036] 1 H NMR (400 MHz, CDCl3) δ 7.61 (dd, J = 7.8, 1.4 Hz, 1H), 7.58 (d, J = 8.1Hz, 2H), 7.46 (dd, J = 7.9, 1.4 Hz, 1H), 7.30 (s, 1H), 7.28 (s, 1H), 6.56 (s,2H), 6.53 (t, J = 7.9 Hz, 1H), 2.46 (s, 3H).

[0037] Synthesis of compound L4a: Dissolve cyanoacetic acid (45 mg, 0.53 mmol) in 12 mL of dichloromethane and stir, then slowly add phosphorus pentachloride (108.5 mg, 0.52 mmol) and stir, slowly add a dichloromethane solution of L3a (150 mg, 0.52 mmol) dropwise at 45 °C to react, after the reaction is completed by TCL detection, add NaOH (60 mg, 1.5 mmol) and stir for 2 hours, quench the reaction with saturated sodium chloride solution, extract with ethyl acetate, collect the organic layer, dry over anhydrous sodium sulfate, remove the solvent by rotary evaporation under reduced pressure, and recrystallize with ethyl acetate to obtain 110 mg of a white solid with a yield of 58%.

[0038] 1 H NMR (400 MHz, CDCl3) δ 9.42 (s, 1H), 7.88 (d, J = 7.7 Hz, 1H), 7.45 –7.38 (m, 3H), 7.35 (d, J = 7.7 Hz, 2H), 7.12 (t, J = 7.9 Hz, 1H), 2.49 (s, 3H).

[0039] Synthesis of compound L5a: Compound L4a (110 mg, 0.325 mmol), benzyl bromide (62 mg, 0.36 mmol) and potassium carbonate (50 mg, 0.36 mmol) were dissolved in 5 mL DMF and stirred at room temperature for 3 h. After the reaction was completed by TCL detection, water was added to quench the reaction, and the mixture was extracted with ethyl acetate and dried over anhydrous sodium sulfate. The solvent was removed by rotary evaporation under reduced pressure, and 54 mg of a white solid was obtained by petroleum ether / ethyl acetate column chromatography with a yield of 39%.

[0040] 1 H NMR (400 MHz, CDCl3) δ 8.08 (d, J = 7.6 Hz, 1H), 7.72 (d, J = 7.0 Hz,2H), 7.60 (d, J = 8.4 Hz, 1H), 7.44 – 7.38 (m, 4H), 7.36 (d, J = 8.1 Hz, 2H),7.27 – 7.21 (m, 1H), 6.86 (d, J = 7.6 Hz, 1H), 5.79 (s, 2H), 2.50 (s, 3H).

[0041] Synthesis of compound S-01: Compound M5a (54 mg, 0.13 mmol), benzylamine (14 mg, 0.13 mmol), Pd2(dba)3 (1.2 mg, 0.0013 mmol), BINAP (1.6 mg, 0.0026 mmol), NaOBu -t (18.7 mg, 0.195 mmol) was dissolved in toluene (6 mL), reacted at 90 °C for 8 h under argon protection, and monitored by TLC. After completion, it was poured into water, washed with saturated brine, extracted with ethyl acetate, and the solvent was removed by rotary evaporation under reduced pressure. 44 mg of a yellow solid was obtained by petroleum ether / ethyl acetate column chromatography. The yield was 74%.

[0042] 1 H NMR (400 MHz, CDCl3) δ 7.51 (d, J = 6.9 Hz, 2H), 7.42 (d, J = 7.4 Hz,4H), 7.39 – 7.31 (m, 8H), 7.19 (t, J = 8.0 Hz, 1H), 6.88 (d, J = 8.1 Hz, 1H),6.75 (d, J = 7.6 Hz, 1H), 6.12 (s, 1H), 5.63 (s, 2H), 4.57 (d, J = 3.5 Hz, 2H),2.49 (s, 3H). 13 C NMR (101 MHz, CDCl3) δ 158.67, 157.81, 143.21, 139.62,138.97, 136.58, 136.01, 131.63, 129.33, 129.19, 128.73, 128.55, 127.92,127.51, 127.34, 127.26, 126.12, 123.28, 115.18, 113.83, 108.78, 97.92, 77.36,77.25, 77.04, 76.73, 68.65, 47.73, 31.44, 30.19, 21.44.

[0043] Example 2

[0044] This example provides compound S-02, whose structural formula and name are as follows: .

[0045] This example provides a method for synthesizing compound S-02, which is different from the method for synthesizing compound S-01 in Example 1 in that the benzylamine in the last step of the compound synthesis is replaced with 1-phenylethyl-1-amine.

[0046] In this example, 34 mg of yellow solid was obtained with a yield of 52%.

[0047] 1 H NMR (400 MHz, CDCl3) δ 7.60 (d, J = 7.4 Hz, 2H), 7.46 – 7.39 (m, 3H), 7.39 – 7.31 (m, 9H), 7.05 (t, J= 8.0 Hz, 1H), 6.80 (d, J = 8.3 Hz, 1H), 6.50 (d, J = 7.6 Hz, 1H), 6.07 (d, J = 4.9 Hz, 1H), 5.71 (s, 2H), 4.83 – 4.40 (m, 1H), 2.47 (s, 3H), 1.70 (d, J = 6.7 Hz, 3H). 13 C NMR (101 MHz, CDCl3) δ 158.68,157.68, 144.66, 142.27, 139.59, 136.87, 135.89, 131.63, 129.32, 129.29,129.18, 129.02, 128.96, 128.70, 128.66, 128.39, 128.21, 127.94, 127.43,127.01, 126.04, 125.73, 123.20, 115.25, 113.60, 109.84, 97.80, 77.36, 77.24,77.04, 76.72, 68.69, 53.22, 25.18, 21.42.

[0048] Example 3

[0049] This example provides compound S-03, whose structural formula and name are as follows: .

[0050] This example provides a method for synthesizing compound S-03. Compared with the method for synthesizing compound S-01 in Example 1, the only difference is that the benzyl group at position N1 of the compound is replaced by a cyclopentyl group, and the benzylamine at position C8 is replaced by 1-(pyridin-2-yl)ethan-1-amine.

[0051] In this example, 57 mg of yellow solid was obtained with a yield of 43%.

[0052] 1 H NMR (400 MHz, CDCl3) δ 8.64 (d, J = 4.6 Hz, 1H), 7.71 – 7.60 (m, 1H), 7.41 (d, J = 7.9 Hz, 1H), 7.36 (d, J = 4.8 Hz, 4H), 7.20 (dd, J= 6.9, 5.3 Hz, 1H),7.09 (t, J = 8.0 Hz, 1H), 6.84 (d, J = 8.3 Hz, 1H), 6.57 (d, J = 7.6 Hz, 1H), 6.51(d, J = 4.9 Hz, 1H), 4.89 – 4.71 (m, 1H), 4.55 (d, J = 6.9 Hz, 2H), 2.59 (dt, J =14.8, 7.5 Hz, 1H), 2.48 (s, 3H), 1.96 (d, J = 6.3 Hz, 2H), 1.81 (d, J = 7.8 Hz,2H), 1.74 (d, J = 6.7 Hz, 3H), 1.71 – 1.60 (m, 2H), 1.53 (dt, J = 19.8, 7.2 Hz,2H). 13 C NMR (101 MHz, CDCl3) δ 163.63, 158.51, 158.38, 149.33, 142.16, 139.47,136.92, 136.16, 131.71, 129.26, 129.17, 129.11, 125.69, 123.04, 122.07,119.86, 115.20, 113.79, 109.48, 97.95, 77.32, 77.20, 77.00, 76.68, 71.00,54.78, 38.70, 29.55, 29.54, 25.44, 23.15, 21.40.

[0053] Example 4

[0054] This example provides compound S-04, whose structural formula and name are as follows: .

[0055] This example provides a method for synthesizing compound S-04, which is different from the method for synthesizing compound S-01 in Example 1 in that the benzyl group at position N1 of the compound is replaced by pyridylmethyl, and the benzylamine at position C8 is replaced by 1-(pyridin-2-yl)ethan-1-amine.

[0056] In this example, 46 mg of yellow solid was obtained with a yield of 66%.

[0057] 1 H NMR (400 MHz, CDCl3) δ 8.65 (t, J = 5.6 Hz, 3H), 7.63 (t, J = 7.7 Hz,1H), 7.56 (d, J = 5.6 Hz, 2H), 7.43 – 7.34 (m, 4H), 7.26 (s, 1H), 7.23 – 7.17(m, 1H), 7.13 (t, J = 8.1 Hz, 1H), 6.86 (d, J = 8.3 Hz, 1H), 6.59 (d, J = 7.7 Hz,1H), 6.31 (s, 1H), 5.74 (s, 2H), 4.75 (d, J = 6.3 Hz, 1H), 2.49 (s, 3H), 1.69(d, J = 6.7 Hz, 3H). 13 C NMR (101 MHz, CDCl3) δ 163.27, 158.95, 157.11, 150.04,149.33, 145.88, 142.17, 139.77, 136.98, 135.75, 131.41, 129.34, 129.16,129.11, 126.38, 123.50, 122.18, 121.60, 119.88, 114.98, 113.82, 109.81,97.57, 77.35, 77.23, 77.03, 76.71, 66.78, 54.55, 23.10, 21.41.

[0058] Example 5

[0059] This example provides compound S-05, whose structural formula and name are as follows: .

[0060] This example provides a method for synthesizing compound S-05, which is different from the method for synthesizing compound S-01 in Example 1 in that the benzylamine at position C8 of the compound is replaced with pyridin-2-ylmethylamine.

[0061] In this example, 47 mg of yellow solid was obtained with a yield of 47%.

[0062] 1 H NMR (400 MHz, CDCl3) δ 8.69 (d, J = 4.4 Hz, 1H), 7.68 (dd, J = 16.8,7.7 Hz, 3H), 7.40 (d, J = 11.2 Hz, 7H), 7.35 – 7.31 (m, 1H), 7.28 – 7.23 (m,1H), 7.20 (t, J = 8.0 Hz, 1H), 6.90 (d, J = 8.4 Hz, 1H), 6.74 (d, J = 7.6 Hz, 1H), 5.73 (s, 2H), 4.72 (s, 2H), 2.49 (s, 3H). 13 C NMR (101 MHz, CDCl3) δ 158.62,158.25, 157.84, 149.29, 143.04, 139.60, 136.79, 136.56, 136.20, 131.63,129.31, 129.18, 128.51, 127.96, 127.81, 126.07, 123.31, 122.24, 121.30,115.17, 113.96, 108.85, 97.96, 77.35, 77.24, 77.04, 76.72, 68.57, 48.94, 21.42.

[0063] Example 6

[0064] This example provides compound S-06, whose structural formula and name are as follows: .

[0065] This example provides a method for synthesizing compound S-06. Compared with the method for synthesizing compound S-01 in Example 1, the only difference is that the toluene group at position C4 of the compound is replaced by isopropyl, and the benzylamine at position C8 is replaced by 1-(pyridin-2-yl)ethan-1-amine.

[0066] In this example, 32 mg of yellow solid was obtained with a yield of 64%.

[0067] 1 H NMR (400 MHz, CDCl3) δ 8.63 (d, J = 4.6 Hz, 1H), 7.61 (dd, J= 12.5,7.5 Hz, 3H), 7.38 (dt, J = 15.6, 7.2 Hz, 4H), 7.26 (d, J = 8.4 Hz, 1H), 7.17 (dd, J = 15.6, 7.6 Hz, 2H), 6.53 (d, J = 7.7 Hz, 1H), 6.38 (d, J = 5.9 Hz, 1H), 5.68(s, 2H), 4.73 (dd, J = 13.0, 6.6 Hz, 1H), 3.96 (dt, J = 14.2, 7.2 Hz, 1H), 1.71(d, J = 6.7 Hz, 3H), 1.61 (d, J = 7.2 Hz, 6H). 13 C NMR (101 MHz, CDCl3) δ 163.69,149.25, 142.78, 136.97, 136.88, 128.56, 127.85, 127.46, 125.61, 122.05,119.69, 109.31, 68.47, 54.74, 23.19.

[0068] Example 7

[0069] This example provides compound S-07, whose structural formula and name are as follows: .

[0070] This example provides a method for synthesizing compound S-07, which is different from the method for synthesizing compound S-01 in Example 1 in that the benzyl group at position N1 of the compound is replaced by a cyclopropylmethyl group, and the benzylamine at position C8 is replaced by 1-(pyridin-2-yl)ethan-1-amine.

[0071] In this example, 39 mg of yellow solid was obtained with a yield of 37%.

[0072] 1 H NMR (400 MHz, CDCl3) δ 8.63 (d, J = 4.8 Hz, 1H), 7.64 (t, J = 7.7 Hz,1H), 7.42 – 7.32 (m, 5H), 7.23 – 7.16 (m, 1H), 7.09 (t,J = 8.0 Hz, 1H), 6.83(d, J = 8.4 Hz, 1H), 6.57 (d, J = 7.7 Hz, 1H), 6.49 (d, J = 5.8 Hz, 1H), 4.95 –4.67 (m, 1H), 4.51 (d, J = 7.1 Hz, 2H), 2.48 (s, 3H), 1.73 (d, J = 6.7 Hz, 3H),1.52 (dt, J = 12.1, 4.8 Hz, 1H), 0.72 (d, J = 7.9 Hz, 2H), 0.53 (d, J = 4.9 Hz,2H). 13 C NMR (101 MHz, CDCl3) δ 163.57, 158.48, 158.36, 149.35, 142.16, 139.49,136.90, 136.14, 131.71, 129.27, 129.17, 129.12, 125.74, 123.05, 122.07,119.86, 115.34, 113.79, 109.51, 97.95, 77.35, 77.03, 76.71, 71.71, 54.74,23.15, 21.40, 9.95, 3.53, 3.50.

[0073] Example 8

[0074] This example provides compound S-08, whose structural formula and name are as follows: .

[0075] This example provides a method for synthesizing compound S-08. Compared with the method for synthesizing compound S-01 in Example 1, the only difference is that the toluyl group at position C4 of the compound is replaced by a cyclohexyl group, and the benzylamine at position C8 is replaced by 1-(pyridin-2-yl)ethan-1-amine.

[0076] In this example, 51 mg of yellow solid was obtained with a yield of 58%.

[0077] 1 H NMR (400 MHz, CDCl3) δ 8.63 (d, J = 4.7 Hz, 1H), 7.61 (dd,J = 14.3,7.3 Hz, 3H), 7.40 (t, J = 7.3 Hz, 2H), 7.37 – 7.31 (m, 1H), 7.26 (d, J = 8.5 Hz,1H), 7.21 – 7.12 (m, 2H), 6.53 (s, 1H), 6.38 (s, 1H), 5.68 (s, 2H), 4.74 (s,1H), 3.53 (d, J = 10.4 Hz, 1H), 2.33 (d, J = 25.2 Hz, 3H), 1.96 (d, J = 8.0 Hz,2H), 1.83 (s, 3H), 1.70 (d, J = 6.7 Hz, 3H), 1.48 (s, 3H). 13 C NMR (101 MHz, CDCl3) δ 163.70, 149.26, 142.74, 136.97, 128.57, 127.84, 127.47, 125.91,122.57, 122.06, 119.72, 109.51, 77.37, 77.06, 76.74, 68.48, 54.75, 26.89, 23.20.

[0078] Example 9

[0079] This example provides compound S-09, whose structural formula and name are as follows: .

[0080] This example provides a method for synthesizing compound S-09. Compared with the method for synthesizing compound S-01 in Example 1, the only difference is that the benzyl group at position N1 of the compound is replaced by 3-chloro-4-methoxybenzyl, and the benzylamine at position C8 is replaced by 1-(pyridin-2-yl)ethan-1-amine.

[0081] In this example, 31 mg of yellow solid was obtained with a yield of 26%.

[0082] 1 H NMR (400 MHz, CDCl3) δ 8.65 (d, J = 4.0 Hz, 1H), 7.64 (dd, J = 16.4,8.8 Hz, 2H), 7.55 (d, J= 8.4 Hz, 1H), 7.40 – 7.31 (m, 5H), 7.23 – 7.18 (m,1H), 7.10 (t, J = 8.0 Hz, 1H), 6.96 (d, J = 8.4 Hz, 1H), 6.83 (d, J = 8.3 Hz, 1H),6.58 (d, J = 7.6 Hz, 1H), 6.44 (s, 1H), 5.65 (s, 2H), 4.79 (d, J = 5.7 Hz, 1H), 3.91 (s, 3H), 2.47 (s, 3H), 1.76 (d, J = 6.7 Hz, 3H). 13 C NMR (101 MHz, CDCl3) δ163.48, 158.73, 157.52, 154.78, 149.31, 142.14, 139.61, 136.96, 135.91,131.54, 129.99, 129.85, 129.28, 129.15, 129.09, 127.62, 126.06, 123.31,122.42, 122.12, 119.83, 115.14, 113.79, 112.08, 109.65, 97.84, 67.58, 56.15,54.63, 23.20, 21.39.

[0083] Example 10

[0084] This example provides a compound S-10, whose structural formula and name are as follows: .

[0085] This example provides a method for synthesizing compound S-10, which is different from the method for synthesizing compound S-01 in Example 1 in that the benzylamine at position C8 of the compound is replaced with an n-butylamine group.

[0086] In this example, 44 mg of yellow solid was obtained with a yield of 69%.

[0087] 1H NMR (400 MHz, CDCl3) δ 7.58 (d, J = 7.5 Hz, 2H), 7.42 (t, J = 7.5Hz, 2H), 7.36 (d, J = 8.8 Hz, 5H), 7.23 (t, J = 8.1 Hz, 1H), 6.84 (d, J = 8.4Hz, 1H), 6.76 (d, J = 7.6 Hz, 1H), 5.65 (s, 2H), 3.31 (dd, J = 11.4, 6.7 Hz, 2H), 2.49 (s, 3H), 1.77 (dt, J = 14.7, 7.2 Hz, 2H), 1.58 – 1.47 (m, 2H), 1.04(t, J = 7.4 Hz, 3H). 13 C NMR (101 MHz, CDCl3) δ 158.63, 157.68, 143.63, 139.55,136.66, 135.91, 131.70, 129.29, 129.17, 128.57, 127.94, 127.46, 126.19,123.30, 115.18, 113.13, 108.11, 97.80, 77.32, 77.01, 76.69, 68.50, 43.18,31.37, 21.41, 20.40, 13.96.

[0088] Embodiment 11

[0089] This example provides compound S-11, whose structural formula and name are as follows: .

[0090] This example provides a method for synthesizing compound S-11, which is different from the method for synthesizing compound S-01 in Example 1 in that the benzyl group at position N1 of the compound is replaced by benzo[d][1,3]dioxolan-5-ylmethyl, and the benzylamine at position C8 is replaced by 1-(pyridin-2-yl)ethan-1-amine.

[0091] In this example, 38 mg of yellow solid was obtained with a yield of 55%.

[0092] 1 H NMR (400 MHz, CDCl3) δ 8.65 (d, J = 4.7 Hz, 1H), 7.63 (td, J = 7.7, 1.6Hz, 1H), 7.34 (dd,J = 10.3, 5.4 Hz, 5H), 7.20 (dd, J = 6.9, 5.4 Hz, 1H), 7.14(s, 1H), 7.10 (dd, J = 14.1, 5.9 Hz, 2H), 6.83 (d, J = 7.8 Hz, 2H), 6.57 (d, J =7.6 Hz, 1H), 6.43 (d, J = 6.3 Hz, 1H), 6.09 – 5.87 (m, 2H), 5.63 (s, 2H), 4.97– 4.58 (m, 1H), 2.47 (s, 3H), 1.75 (d, J = 6.7 Hz, 3H). 13 C NMR (101 MHz, CDCl3)δ 163.58, 158.64, 157.66, 149.34, 147.85, 147.41, 142.18, 139.57, 136.93,135.99, 131.59, 130.54, 129.28, 129.10, 125.98, 123.25, 122.08, 121.62,119.77, 115.18, 113.77, 109.59, 108.60, 108.24, 101.08, 77.31, 76.99, 76.68,68.58, 54.69, 23.16, 21.39.

[0093] Example 12

[0094] This example provides compound S-12, whose structural formula and name are as follows: .

[0095] This example provides a method for synthesizing compound S-12, which is different from the method for synthesizing compound S-01 in Example 1 in that the benzyl group at position C4 of the compound is replaced by 4-methoxyphenyl, and the benzylamine at position C8 is replaced by R -1-(Pyridin-2-yl)ethan-1-amine.

[0096] In this example, 45 mg of yellow solid was obtained with a yield of 69%.

[0097] 1 H NMR (400 MHz, CDCl3) δ 8.64 (d, J= 4.1 Hz, 1H), 7.63 (dd, J = 18.0,7.5 Hz, 3H), 7.41 (s, 4H), 7.37 – 7.31 (m, 2H), 7.23 – 7.16 (m, 1H), 7.09 (t, J = 8.0 Hz, 3H), 6.86 (d, J = 8.3 Hz, 1H), 6.56 (d, J = 7.6 Hz, 1H), 6.39 (s, 1H), 5.74 (s, 2H), 4.78 (s, 1H), 3.91 (s, 3H), 1.73 (d, J = 6.5 Hz, 3H). 13 C NMR (101MHz, CDCl3) δ 163.61, 160.55, 158.28, 157.76, 149.26, 142.20, 137.01, 136.83,136.01, 130.83, 130.77, 128.59, 127.92, 127.61, 126.63, 125.99, 123.34,122.10, 119.76, 115.33, 114.05, 113.75, 109.60, 97.82, 68.63, 55.33, 54.72,23.19.

[0098] Embodiment 13

[0099] In this example, the PDE5 enzyme inhibitory activity of compounds S-01 to S-12 provided in Examples 1 to 12 was determined.

[0100] The assay includes the following steps: (1) Drug preparation: Prepare a 10 mM stock solution with DMSO, dissolve it by ultrasound, and then perform gradient dilution with DMSO and kinase buffer to ensure that the final concentration of DMSO is less than 1%.

[0101] (2) Composition of kinase reaction buffer: 40 mM MOPS, pH 7.5, 0.5 mM EDTA, 15 mM MgCl2, 0.15 mg / ml BSA, 1 mM DTT, 0.05% Proclin 200, 15 ng / ml PDE 4CAT, 100 nM FAM-Cyclic-3',5'-AMP.

[0102] (3) The detailed experimental steps are as follows: At 25°C, the compound was diluted with 10% DMSO and 5 µl of the dilution was added to a 50 µl reaction mixture to make the final DMSO concentration of all reactions 1%. 5 µl of the diluted drug was added to a 50 µl reaction mixture system, which contained 40 mM MOPS, pH 7.5, 0.5 mM EDTA, 15 mMMgCl2, 0.15 mg / ml BSA, 1 mM DTT, 0.05% Proclin 200, 15 ng / ml PDE 4CAT and 100 nM FAM-Cyclic-3',5'-AMP. The reaction mixture was incubated at 25°C for 1 hour. Then, 100 ul of the diluted binding agent was added to each well and incubated at 25°C for 1 hour with slow shaking. The fluorescence polarization of the samples was read using a 360 nm excitation filter and a 480 nm emission filter. Data were collected and the IC values ​​of the compounds were calculated using Prism GraphPad software. 50 value.

[0103]

[0104] Experimental conclusion: As shown in Table 1, most of the compounds of the present invention have a strong inhibitory effect on PDE5 protease, and the IC values ​​of in vitro enzyme inhibition activity are 50 There are 4 compounds below 20 nM, namely compounds S-01, S-02, S-05 and S-12. In particular, compound S-12 has an IC of 2.17 for in vitro protein inhibition of PDE5. 50 The positive control drug used in the activity test of the present invention is sildenafil, and its IC value for PDE5 inhibitory activity under the same test conditions is 50 is 4 nM.

[0105] Embodiment 14

[0106] In this example, an animal experiment was conducted on the compound S-12 provided in Example 12 to study its effect on the rat pulmonary hypertension model induced by monocrotaline.

[0107] The experimental plan is as follows: The pharmacodynamic effect of S-12 on PAH was evaluated in a rat model of PAH induced by monocrotaline (MCT). Experimental plan: 47 male SPF SD rats weighing 180-220g were randomly divided into 4 groups, namely, vehicle control group, model control group, S-12 (5.0 mg / kg), and sildenafil positive control group (10.0 mg / kg). Due to animal death in the late stage of the model, according to previous experience, except for 8 rats in each vehicle control group, the remaining groups were designed to have 13 rats in each group. Except for the vehicle control group, the remaining groups were intraperitoneally injected with 2% monocrotaline 60 mg / kg to establish the model, and the drug was administered the next day for 21 consecutive days. After the end of the drug administration, the mean pulmonary artery pressure (mPAP) and right ventricular hypertrophy index (RVHI%) of each group were measured. The wall thickness percentage (WT%) was observed by HE staining.

[0108] The experimental results are shown in Figure 1 and Figure 2 .

[0109] Figure 1 and Figure 2 The results showed that compared with the vehicle control group, the mPAP in the model group was significantly increased. Compared with the model control group, both S-12 and the positive drug sildenafil could improve the above-mentioned typical symptoms of PAH, and S-12 achieved better efficacy at a lower dosage, further demonstrating the feasibility of PDE5 inhibitors for anti-PAH treatment.

[0110] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A dihydroquinolinone compound, characterized in that: It has a chemical structure as shown in the general formula (I): （Ⅰ） In the general formula (I), R 1 is C3~C6 alkyl or substituted C3~C6 cycloalkyl, heteroaryl or benzobisoxazolyl; R 2 is phenyl or substituted phenyl, C1~C6 alkyl or C3~C6 cycloalkyl; R 3 is C1~C5 alkyl, phenyl or substituted phenyl, pyridyl, or halogenated pyridyl; The substituent of the substituted phenyl group is C1-C5 alkyl, methoxy, or halogen.

2. The dihydroquinolinone compound according to claim 1, characterized in that R 1 Select from any of the following structures: 。 3. The dihydroquinolinone compound according to claim 1, characterized in that R 2 Select from any of the following structures: 。 4. The dihydroquinolinone compound according to claim 1, characterized in that R 3 Select from any of the following structures: 。 5. The dihydroquinolinone compound according to claim 1, characterized in that It has any of the following structures: ； ； ； ； ； ； ； ； ； ； ； 。 6. The pharmaceutically acceptable salt or stereoisomer of the dihydroquinolinone compound according to any one of claims 1 to 5.

7. A phosphodiesterase 5 activity inhibitor, characterized in that The main active ingredient is the dihydroquinolinone compound according to any one of claims 1 to 5 or the salt or stereoisomer according to claim 6.

8. Use of the dihydroquinolinone compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt or stereoisomer thereof, or the phosphodiesterase 5 activity inhibitor according to claim 7 in the preparation of a medicament for treating phosphodiesterase 5-related diseases.

9. The use according to claim 8, characterized in that: The medicament is used to treat pulmonary hypertension.

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