Dihydroquinolinone compounds and uses thereof

Abstract

The application relates to the technical field of biological medicine, in particular to a new skeleton dihydroquinolinone compound and application thereof. The application provides an 8-substituted dihydroquinolinone compound with a structure as shown in formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof and application thereof. The 8-substituted dihydroquinolinone compound has significant inhibitory activity on phosphodiesterase 5 (PDE5) type, has significant therapeutic effect on a phosphodiesterase 5 related disease 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] This invention relates to the field of biomedical technology, and in particular to an 8-substituted dihydroquinolinone compound and its applications. Background Technology

[0002] Pulmonary artery hypertension (PAH) is a malignant pulmonary vascular disease characterized by progressive pulmonary artery occlusion leading to gradually increasing pulmonary vascular resistance and ultimately right ventricular failure (RVF). Its prominent pathological feature is intimal proliferation and remodeling of the pulmonary arterioles. The 5-year survival rate for PAH patients is only about 34%, hence its nickname "cancer of cardiovascular diseases." Although targeted drug therapy for PAH has made significant progress in recent years in improving clinical observation indicators (exercise tolerance, hemodynamic parameters, etc.) and delaying the time to clinical deterioration, long-term prognosis remains unsatisfactory. Currently used first-line anti-PAH drugs can only slow the progression of the disease and cannot achieve a clinically effective cure. In-depth exploration of the pathogenesis of pulmonary artery hypertension and the development of new targets that may intervene in pulmonary artery hypertension are of great significance for new drug development and disease treatment.

[0003] Phosphodiesterases (PDEs) are a superfamily of intracellular enzymes that primarily regulate cell signal transduction by catalyzing the degradation of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP). Cyclic nucleotides (cAMP and cGMP) serve as important intracellular second messengers, mediating and participating in various physiological regulations and the development of diseases. Adenylate cyclase (AC) and guanylate cyclase (GC) synthesize cAMP and cGMP molecules and degrade them through PDEs, maintaining a delicate balance in intracellular cAMP and cGMP levels. cAMP mainly functions through protein kinase A (PKA) and cAMP-activated exchangers (EPACs), regulating events such as cell movement, migration, hormone secretion, cell metabolism, and gene transcription. Human PDEs comprise 11 subtypes, among which PDE5 specifically hydrolyzes cGMP and is a key regulator of cGMP signaling, primarily distributed in platelets and vascular smooth muscle. It participates in the NO-cGMP signaling pathway in platelets, thereby regulating cGMP signaling in the brain and modulating vascular smooth muscle contraction, particularly in the penis and lungs. Currently, sildenafil and tadalafil are classic marketed PDE5 inhibitors used to treat PAH. They increase intracellular cGMP concentration by inhibiting the hydrolytic function of PDE5, thereby exerting vasodilatory effects and inhibiting vascular endothelial cell proliferation, thus achieving anti-PAH efficacy. However, existing studies have found that sildenafil has poor pharmacokinetic stability, a short half-life, and long-term use can easily cause muscle pain and visual disturbances. Therefore, discovering highly active and selective PDE5 inhibitors and leveraging their important role in the research of novel anti-PAH target drugs will bring new opportunities for drug treatment of PAH patients.

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

[0005] To address the aforementioned technical problems, this invention provides a novel dihydroquinoline ketone compound and its applications. The dihydroquinoline ketone compound exhibits significant inhibitory activity against phosphodiesterase type 5 and can be used to prepare novel target drug candidates for pulmonary hypertension.

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

[0007] (I)

[0008] In general formula (I),

[0009] R 1It is a C3-C6 alkyl group or a substituted C3-C6 cycloalkyl group, heteroaryl group or benzodioxazolyl group; R 2 It is a phenyl or substituted phenyl, a C1-C6 alkyl or a C3-C6 cycloalkyl; R 3 It is a C1~C5 alkyl, phenyl or substituted phenyl, pyridyl or halopyridyl; The substituents of the substituted phenyl group are C1-C5 alkyl, methoxy, or halogen.

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

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

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

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

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

[0015] Secondly, 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] Thirdly, the present invention provides a phosphodiesterase 5 activity inhibitor, the main active ingredient of which includes the aforementioned dihydroquinolinone compounds or their pharmaceutically acceptable salts or stereoisomers.

[0018] Fourthly, the present invention provides the use of the aforementioned dihydroquinolinone compounds, or pharmaceutically acceptable salts or stereoisomers thereof, or the phosphodiesterase 5 activity inhibitors in the preparation of medicaments for treating phosphodiesterase 5-related diseases.

[0019] Preferably, the drug is used to treat pulmonary hypertension.

[0020] Beneficial effects: This invention provides an 8-substituted dihydroquinolinone compound having the structure shown in Formula (I), or a pharmaceutically acceptable salt or stereoisomer thereof, and its applications. The dihydroquinolinone compound of this invention exhibits significant inhibitory activity against phosphodiesterase 5 (PDE5) and demonstrates significant therapeutic effects on phosphodiesterase 5-related diseases, such as animal models of pulmonary hypertension, without significant side effects. It can be used to prepare novel target drug candidates for pulmonary hypertension. Attached Figure Description

[0021] To more clearly illustrate the technical solutions in this invention or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be described below.

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

[0023] Figure 2 The results of HE staining were obtained to observe the percentage of wall thickness (WT%) of the chamber 21 days after administration of compound S-12 (5.0 mg / kg) in Example 14 of this invention. Detailed Implementation

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

[0025] The compounds provided by this invention have the chemical structure shown in general formula (I):

[0026] (I)

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

[0028] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this 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 the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0030] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "specific implementation," or "some specific implementations," etc., refer to specific features, structures, materials, or characteristics described in connection with that embodiment or example, which are included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0031] In the embodiments provided in this specification, unless specific techniques or conditions are specified, the techniques or conditions described in the literature in this field, or the product instructions, shall be followed. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased from legitimate channels.

[0032] Example 1

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

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

[0035] in: Synthesis of compound L3a: In a 100 mL reaction flask, L2a (985 mg, 5 mmol) was added and dissolved in 20 mL THF. Under ice bath conditions, 15 mL of p-tolyl magnesium bromide was added dropwise to the system using a constant pressure funnel. After the addition was complete, the mixture was refluxed for 8 h. The reaction was monitored by thin-layer chromatography (TLC). After the reaction was complete, 6 mL of 4 mol / L hydrochloric acid was slowly added dropwise to the above reaction system, and the reaction was allowed to proceed overnight. After the reaction was confirmed by TLC, the solvent was removed under reduced pressure. The pH of the solution was adjusted to neutral with saturated sodium bicarbonate solution. The mixture was extracted twice with dichloromethane, and the organic layer was collected, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. After drying, the solution was subjected to column chromatography (petroleum ether: ethyl acetate = 20:1) to give 1.05 g of a yellow oil, 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: Cyanoacetic acid (45 mg, 0.53 mmol) was dissolved in 12 mL of dichloromethane and stirred. Phosphorus pentachloride (108.5 mg, 0.52 mmol) was then slowly added and stirred. A dichloromethane solution of L3a (150 mg, 0.52 mmol) was slowly added dropwise at 45 °C. After the reaction was completed as detected by TCL, NaOH (60 mg, 1.5 mmol) was added and stirred for 2 hours. The reaction was quenched with saturated sodium chloride solution, extracted with ethyl acetate, and the organic layer was collected, dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation under reduced pressure. Recrystallization with ethyl acetate gave 110 mg of white solid, yield 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 of DMF and stirred at room temperature for 3 h. After the reaction was detected by TCL, the reaction was quenched with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation under reduced pressure. The white solid was obtained by column chromatography in petroleum ether / ethyl acetate, yielding 54 mg, 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), and NaOBu were prepared by mixing these components. -t (18.7 mg, 0.195 mmol) was dissolved in toluene (6 mL), and reacted at 90 °C for 8 h under argon protection. The reaction was monitored by TLC. After the reaction was completed, the mixture was poured into water, washed with saturated brine, extracted with ethyl acetate, and the solvent was removed by rotary evaporation under reduced pressure. The solid was then purified by column chromatography with petroleum ether / ethyl acetate to give 44 mg of yellow solid, with a yield of 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 embodiment provides compound S-02, whose structural formula and name are as follows: .

[0045] This embodiment provides a method for synthesizing compound S-02. Compared with the method for synthesizing compound S-01 in Example 1, the only difference is that 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 embodiment provides compound S-03, whose structural formula and name are as follows: .

[0050] This embodiment 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 the N1 position of the compound is replaced with a cyclopentyl group, and the benzylamine at the C8 position is replaced with 1-(pyridin-2-yl)ethyl-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 embodiment provides compound S-04, whose structural formula and name are as follows: .

[0055] This embodiment provides a method for synthesizing compound S-04. Compared with the method for synthesizing compound S-01 in Example 1, the only difference is that the benzyl group at the N1 position of the compound is replaced with pyridylmethyl, and the benzylamine at the C8 position is replaced with 1-(pyridin-2-yl)ethyl-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 embodiment provides compound S-05, whose structural formula and name are as follows: .

[0060] This embodiment provides a method for synthesizing compound S-05. Compared with the method for synthesizing compound S-01 in Example 1, the only difference is that the benzylamine at the C8 position of the compound is replaced with pyridin-2-methylamine.

[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 embodiment provides compound S-06, whose structural formula and name are as follows: .

[0065] This embodiment 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 tolyl group at the C4 position of the compound is replaced with isopropyl, and the benzylamine at the C8 position is replaced with 1-(pyridin-2-yl)ethyl-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 embodiment provides compound S-07, whose structural formula and name are as follows: .

[0070] This embodiment provides a method for synthesizing compound S-07. Compared with the method for synthesizing compound S-01 in Example 1, the only difference is that the benzyl group at the N1 position of the compound is replaced with cyclopropylmethyl, and the benzylamine at the C8 position is replaced with 1-(pyridin-2-yl)ethyl-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 embodiment provides compound S-08, whose structural formula and name are as follows: .

[0075] This embodiment 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 tolyl group at the C4 position of the compound is replaced with a cyclohexyl group, and the benzylamine at the C8 position is replaced with 1-(pyridin-2-yl)ethyl-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 embodiment provides compound S-09, whose structural formula and name are as follows: .

[0080] This embodiment 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 the N1 position of the compound is replaced with 3-chloro-4-methoxybenzyl, and the benzylamine at the C8 position is replaced with 1-(pyridin-2-yl)ethyl-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 embodiment provides compound S-10, whose structural formula and name are as follows: .

[0085] This embodiment provides a method for synthesizing compound S-10. Compared with the method for synthesizing compound S-01 in Example 1, the only difference is that the benzylamine at the C8 position of the compound is replaced with a 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] Example 11

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

[0090] This embodiment provides a method for synthesizing compound S-11. Compared with the method for synthesizing compound S-01 in Example 1, the only difference is that the benzyl group at the N1 position of the compound is replaced with benzo[d][1,3]dioxolane-5-ylmethyl, and the benzylamine at the C8 position is replaced with 1-(pyridin-2-yl)ethyl-1-amine.

[0091] In this example, 38 mg of a 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 embodiment provides compound S-12, whose structural formula and name are as follows: .

[0095] This embodiment provides a method for synthesizing compound S-12. Compared to the method for synthesizing compound S-01 in Example 1, the only difference is that the benzyl group at the C4 position is replaced with 4-methoxyphenyl, and the benzylamine at the C8 position is replaced with... R -1-(pyridin-2-yl)ethyl-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] Example 13

[0099] This embodiment measures the PDE5 enzyme inhibitory activity of compounds S-01 to S-12 provided in Examples 1 to 12.

[0100] The determination experiment includes the following steps: (1) Preparation of the drug: Prepare a 10 mM stock solution with DMSO, accelerate the dissolution by sonication, 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 were as follows: At 25°C, the compound was diluted with 10% DMSO, and 5µl of the diluent was added to 50µl of the reaction mixture to make the final concentration of DMSO in all reactions 1%. 5µl of the diluted drug was added to 50µl of the reaction mixture system. The entire reaction system contained 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, and 100 nM FAM-Cyclic-3',5'-AMP. The reaction mixture was incubated at 25°C for 1 hour. Then, 100 µl of the diluted binder was added to each well, and the mixture was incubated at 25°C with slow shaking for 1 hour. The fluorescence polarization of the samples was read using a 360 nm excitation filter and a 480 nm emission filter. Data was collected, and the IC50 of the compounds was calculated using Prism GraphPad software. 50 value.

[0103]

[0104] Experimental conclusion: As shown in Table 1, most of the compounds of this invention have strong inhibitory effects on PDE5 protease, and the IC50 of their in vitro enzymatic inhibitory activity is [data missing]. 50 Four compounds were found to have an IC50 value below 20 nM: compounds S-01, S-02, S-05, and S-12. Compound S-12, in particular, showed a high IC50 value for in vitro protein inhibitory activity against PDE5. 50 The activity level reached 3 nM. The positive control drug used in the activity test of this invention was sildenafil, and its IC50 inhibitory activity against PDE5 was [value missing] under the same test conditions. 50 It is 4 nM.

[0105] Example 14

[0106] This embodiment uses compound S-12 provided in Example 12 to conduct animal experiments to study its effect on a rat pulmonary hypertension model induced by lily alkaloids.

[0107] The specific experimental design is as follows: The pharmacodynamic effects of S-12 on PAH were evaluated in a rat model induced by methyl limonene (MCT). Experimental protocol: Forty-seven male SPF-grade SD rats, weighing 180-220g, were randomly divided into four groups: a solvent control group, a model control group, S-12 (5.0 mg / kg), and a sildenafil positive control group (10.0 mg / kg). Due to potential animal mortality in the later stages of the model, based on past experience, each group except the solvent control group (n=8) was designed with 13 rats per group. Except for the solvent control group, all other groups received an intraperitoneal injection of 2% methyl limonene 60 mg / kg to induce PAH. Drug administration began the following day and continued for 21 days. After drug administration, mean pulmonary artery pressure (mPAP) and right ventricular hypertrophy index (RVHI%) were measured in each group. The percentage of ventricular wall thickness (WT%) was also observed using HE staining.

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

[0109] Figure 1 and Figure 2 The results showed that mPAP levels in the model group were significantly higher than those in the solvent control group. Compared with the model group and the control group, both S-12 and the positive control drug sildenafil could improve the typical PAH symptoms. Moreover, S-12 achieved better efficacy with a lower dosage, further demonstrating the feasibility of using 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, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A dihydroquinoline ketone compound, characterized in that, It has any of the following structures: ； ； ； ； ； ； ； ； ； 。 2. A pharmaceutically acceptable salt of the dihydroquinolinone compound of claim 1.

3. A phosphodiesterase 5 activity inhibitor, characterized in that, Its main active ingredient is the dihydroquinoline ketone compound of claim 1 or the salt of claim 2.

4. The use of the dihydroquinolinone compound of claim 1, or a pharmaceutically acceptable salt thereof, or the phosphodiesterase 5 activity inhibitor of claim 3 in the preparation of a medicament for the treatment of pulmonary hypertension.

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