A dihydroquinolinone derivative, preparation method and application thereof

Abstract

The application belongs to the technical field of biological medicine, and particularly relates to a dihydroquinolinone derivative, a preparation method and application. The dihydroquinolinone derivative has a significant inhibitory effect on phosphodiesterase type 5 (PDE5), and can selectively inhibit PDE5 and has no or extremely weak inhibitory effect on other subtypes PDE. The dihydroquinolinone derivative has a significant therapeutic effect on diseases such as pulmonary fibrosis related to PDE5, has no obvious toxic side effect, is high in safety, and is very suitable for treating PDE5 related diseases.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine. More particularly, it relates to a dihydroquinolinone derivative, a preparation method and an application. BACKGROUND

[0002] The phosphodiesterase (PDEs) superfamily of enzymes is responsible for the hydrolysis of the cellular second messengers cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP). This PDE is encoded by 21 identified genes and can be divided into 11 subfamilies (PDE1-11). Except for PDE6 which is specifically distributed in the rod cells of the retina, each of the other subfamilies is distributed at different levels in different tissues or cells, and its inhibitors can regulate the concentration of cAMP and cGMP in cells by hydrolyzing cAMP and cGMP into 5'AMP and 5'GMP, activate PKA and PKG and downstream proteins to mediate various physiological processes, including: cell growth, mitotic occurrence, glycogenolysis, insulin secretion, etc.

[0003] Among them, PDE5 catalyzes the hydrolysis of the phosphodiesterase bond in cGMP through its catalytic domain, thereby converting cGMP into inactive 5'-GMP. In view of the key role of cGMP in various physiological processes by activating this cellular signaling pathway, PDE5 inhibitors have been shown to have good therapeutic effects on various diseases, such as erectile dysfunction and pulmonary arterial hypertension; also used for the treatment of mental diseases such as Alzheimer's disease, such as sildenafil has completed a phase IV trial of schizophrenia and a phase IV trial of Parkinson's disease, a phase II clinical trial of tadalafil is being conducted in patients with cerebral small vessel disease to determine whether tadalafil can improve blood flow in deep brain tissue and possibly improve cognitive function.

[0004] At present, the U.S. Food and Drug Administration (FDA) has approved four PDE5 inhibitors for marketing, namely sildenafil (related Chinese application CN1253561A), vardenafil, tadalafil and avanafil, mainly for the treatment of male erectile dysfunction (ED). However, the above-mentioned PDE5 inhibitor drugs on the market all have adverse reactions such as headache, flushing, indigestion and visual impairment during use, and have low safety; this is because PDE5 has a high similarity with the PDE6 protein pocket, making it difficult to obtain high subtype selectivity, resulting in most of the existing PDE5 inhibitors having clinical side effects such as visual abnormalities. SUMMARY

[0005] The technical problem to be solved by the present application is to overcome the defects and deficiencies of the existing PDE5 inhibitors with low selectivity and certain safety hazards, and to provide a dihydroquinolinone derivative which has good selective activity for PDE5 and good safety.

[0006] The present application aims to provide a preparation method of the dihydroquinolinone derivative.

[0007] Another object of the present application is to provide the use of the dihydroquinolinone derivative in the preparation of a phosphodiesterase type V inhibitor.

[0008] Another object of the present application is to provide the use of the dihydroquinolinone derivative in the preparation of a drug for treating a phosphodiesterase type V related disease.

[0009] The above objects of the present application are achieved by the following technical solutions.

[0010] A dihydroquinolinone derivative, having the structure of formula (I):

[0011]

[0012] wherein R1, R2, R3, R4, R6, R8 and R9 are each independently selected from hydrogen, halogen, cyano, nitro, hydroxyl, amino, thiol, carboxyl, C1-6 alkyl, halogenated C1-6 alkyl, C3-6 cycloalkyl, C1-6 alkoxy, C1-6 alkanoyl, C1-6 amido;

[0013] R5 is selected from one of hydrogen, -NH-CO-Ra, -NH-CH2-Ra, -NH-CH(CH3)-Ra, and Ra is selected from one of C1-6 alkyl, C3-6 cycloalkyl, phenyl or substituted phenyl, and the substituent of the substituted phenyl is halogen or halogenated C1-6 alkoxy;

[0014] R7 is selected from one of hydrogen, hydroxyl, halogen, cyano, nitro, amino, thiol, sulfate, phosphate, C1-6 carboxylic acid group, C2-6 ester group, C1-6 alkoxy, C1-6 alkyl, halogenated C1-6 alkyl, C3-6 cycloalkyl, C1-6 carboxylic acid substituted C1-6 alkoxy, C2-6 ester group substituted C1-6 alkoxy, C3-6 alkenoic acid group, oxadiazole group, C1-6 alkyl substituted oxadiazole group, hydroxamic acid group, N'-acetylcarbohydrazide group.

[0015] Preferably, R1, R2, R3, R4, R6, R8 and R9 are each independently selected from one or more of hydrogen, halogen, C1-6 alkoxy, hydroxyl, amino;

[0016] R5 is selected from one of hydrogen, -NH-CO-Ra, -NH-CH2-Ra, -NH-CH(CH3)-Ra, and Ra is selected from one of C1-3 alkyl, C3-6 cycloalkyl, phenyl or substituted phenyl, and the substituent of the substituted phenyl is halogen or halogenated C1-3 alkoxy;

[0017] R7is selected from one of hydrogen, hydroxyl, cyano, C2-6 ester, C1-3 alkoxy, C1-3 carboxylic acid substituted C1-3 alkoxy, C2-4 ester substituted C1-3 alkoxy, C3-6 alkenyl, oxadiazolyl, C1-3 alkyl substituted oxadiazolyl, hydroxamic acid, N'-acetylcarbohydrazide.

[0018] More preferably, R1, R2, R3, R4, R6, R8and R9are each independently selected from one or more of hydrogen, chloro, methoxy, hydroxyl, amino;

[0019] R5is selected from one of hydrogen, -NH-CO-Ra, -NH-CH2-Ra, -NH-CH(CH3)-Ra, and Rais methyl, cyclohexyl, phenyl, fluorophenyl, trifluoromethoxyphenyl or piperonyl;

[0020] R7is selected from one of hydrogen, methoxy, hydroxyl, cyano, acetoxyhydroxamic acid, N'-acetylcarbohydrazide, 2-methyloxadiazole, 2-methyloxalate, 2-oxoacetic acid, acrylic acid.

[0021] Further, the dihydroquinolinone derivative also includes a pharmaceutically acceptable salt or solvate thereof.

[0022] In addition, the present application also claims a preparation method of the dihydroquinolinone derivative, when R7is acrylic acid, the synthetic route is as follows:

[0023]

[0024] i. Compound A is reacted with compound B in the presence of a base to form compound C (preferably, the reaction time is 2-12 h);

[0025] ii. Compound C is reacted with compound D in the presence of an acid to form compound E (preferably, the reaction time is 24-48 h);

[0026] iii. Compound E is reacted with a reducing agent to form compound F (preferably, the reaction time is 12-48 h);

[0027] iv. Compound F is reacted with N,N-dimethylformamide, a base and an acrylic ester in the presence of a catalyst to form compound G (preferably, the reaction time is 6-24 h);

[0028] v. Compound G is reacted with compound H in the presence of a reducing agent to form compound I (preferably, the reaction time is 0.5-6 h);

[0029] vi. Compound I is reacted in the presence of a base to form compound J (preferably, the reaction time is 3-6 h);

[0030] wherein R10 is hydrogen, halogen, hydroxyl or methyl; X is -CO-Ra, -CH2-Ra or -CH(CH3)-Ra; R1-R9, Ra are defined as above.

[0031] Further, in steps i, iv, the base is selected from one or more of diisopropylethylamine, triethylamine, 4-dimethylaminopyridine, piperidine, sodium bicarbonate, sodium carbonate, potassium carbonate, sodium hydroxide, potassium hydroxide, potassium tert-butoxide, sodium hydride, sodium methoxide, sodium ethoxide.

[0032] Still further, in step ii, the acid is selected from one or more of hydrochloric acid, sulfuric acid, hydrogen chloride, 4M hydrogen chloride in dioxane, 4M hydrogen chloride in methanol.

[0033] Further, in steps iii, v, the reducing agent is selected from one or more of stannous chloride, ferrous sulfate, Raney nickel, palladium on carbon, lithium aluminum hydride, diisobutylaluminum hydride, lithium borohydride, lithium triethylborohydride, nickel borohydride, zinc borohydride, sodium borohydride, sodium cyanoborohydride, sodium triacetoxyborohydride, lithium tri-sec-butylborohydride, diisopropylaluminum hydride, borane.

[0034] Still further, in step iv, the catalyst is selected from one or more of palladium acetate, palladium chloride, tetrakis(triphenylphosphine)palladium, allylpalladium(II) chloride dimer, dichlorobis(triphenylphosphine)palladium, tris(dibenzylideneacetone)dipalladium(0), bis(tri-t-butylphosphine)palladium(0), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium.

[0035] In addition, the present application also claims the use of the dihydroquinolinone derivative in the preparation of phosphodiesterase type V inhibitors.

[0036] Based on this, the present application also claims the use of the dihydroquinolinone derivative in the preparation of drugs for treating phosphodiesterase type V related diseases.

[0037] Further, the phosphodiesterase type V related diseases include erectile dysfunction, pulmonary arterial hypertension, organ fibrosis, female sexual dysfunction, premature birth, dysmenorrhea, benign prostatic hyperplasia, tumor multidrug resistance, bladder outlet obstruction, incontinence, unstable and variant angina, hypertension, congestive heart failure, renal failure, atherosclerosis, stroke, peripheral vascular disease, Raynaud's disease, inflammatory diseases, bronchitis, chronic asthma, allergic asthma, allergic rhinitis, glaucoma, diseases related to intestinal motility disorders.

[0038] The present application has the following beneficial effects:

[0039] The present application provides a dihydroquinolinone derivative, which has significant inhibitory effect on PED5 and can selectively inhibit PDE5 without or with extremely weak inhibition on other subtypes of PDE; the dihydroquinolinone derivative has significant therapeutic effect on diseases related to PDE5 such as pulmonary fibrosis, and has no obvious toxic side effects, high safety, and is very suitable for treating PDE5 related diseases. BRIEF DESCRIPTION OF DRAWINGS

[0040] Figure 1 Tendency chart of body weight of rats for acute toxicity experiment of compound 22.

[0041] Figure 2 Statistical chart of lung respiratory function index data of experimental rats.

[0042] Figure 3 H&E stained lung tissue section chart of experimental rats.

[0043] Figure 4 Masson stained lung tissue section chart and collagen area ratio chart of experimental rats.

[0044] Figure 5 Western blot chart and data statistical chart of α-SMA protein expression level of experimental rats. DETAILED DESCRIPTION

[0045] The present application will be further described below in combination with the drawings and specific examples, but the examples do not limit the present application in any form. Unless otherwise specified, the reagents, methods and devices used in the present application are conventional reagents, methods and devices in the technical field.

[0046] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0047] Example 1 Synthesis of compound 1

[0048]

[0049] A mixture of dimethyl amino terephthalate (500 mg, 2.39 mmol), 3-chlorophenyl acetonitrile (750 mg, 4.78 mmol) and 15 mL of 4N hydrogen chloride dioxane solution was stirred at 100℃ for 48 hours. Then ice was added, and the PH was adjusted with sodium hydroxide solution, and the precipitated crystals were collected by filtration and washed with water and ethyl acetate in turn. After drying, 470 mg of white solid was obtained with a yield of 51%.

[0050] 1 H NMR (400 MHz, DMSO-D 6)δ12.63(s,1H),8.20(d,J=8.2Hz,1H),8.08(s,1H),7.96(d,J=8.2Hz,1H),7.49(s,1H),7.36(t,J=5.8Hz,3H),3.99(s,2H),3.91(s,3H). 13 C NMR (101MHz, DMSO-D) 6 )δ165.89,161.76,157.08,149.12,139.07,135.15,133.43,130.75,1 29.43,128.46,128.28,127.33,127.01,126.15,124.48,53.11,53.03.

[0051] Example 2: Synthesis of Compound 2

[0052]

[0053] Following the synthetic method of compound 1, 3-chlorophenylacetonitrile was replaced with 3,4-dichlorophenylacetonitrile to obtain a white solid in 50% yield.

[0054] 1 H NMR (400MHz, DMSO-d) 6 )δ12.61(s,1H),8.19(d,J=8.2Hz,1H),8.06(d,J=1.2Hz,1H),7.95(dd,J=8.2,1.6Hz,1H),7.68 (d,J=1.9Hz,1H),7.60(d,J=8.3Hz,1H),7.38(dd,J=8.3,2.0Hz,1H),4.00(s,2H),3.91(s,3H). 13 C NMR (126MHz, DMSO-d) 6 )δ165.40,161.29,156.34,148.58,137.14,134.66,131.24,130.86,130.52,129.62,127.98,126.55,125.74,124.04,52.63.

[0055] Example 3: Synthesis of Compound 3

[0056]

[0057] Following the synthetic method of compound 1, 3-chlorophenylacetonitrile was replaced with 3,4-dimethoxyphenylacetonitrile to obtain a white solid in 50% yield.

[0058] 1H NMR (400 MHz, DMSO-d 6 ) δ 12.56 (s, 1H), 8.19 (d, J = 8.2 Hz, 1H), 8.10 (d, J = 1.2 Hz, 1H), 7.95 (dd, J = 8.2, 1.6 Hz, 1H), 7.05 (s, 1H), 6.90 (s, 2H), 3.91 (s, 3H), 3.88 (s, 2H), 3.75 (s, 3H), 3.71 (s, 3H). 13 C NMR (126 MHz, DMSO-d 6 ) δ 165.64, 161.18, 159.70, 149.10, 148.52, 135.59, 127.44, 127.05, 125.59, 123.87, 121.73, 113.60, 112.37, 66.80, 55.98, 55.96, 53.29.

[0059] Synthesis of compound 4 of example 4

[0060]

[0061] Following the synthesis procedure of compound 1, amino terephthalic acid dimethyl ester was replaced by methyl anthranilate to give a white solid in 30% yield.

[0062] 1 H NMR (400 MHz, DMSO-d 6 ) δ 12.41 (s, 1H), 8.10 - 8.05 (m, 1H), 7.80 - 7.74 (m, 1H), 7.60 (d, J = 8.0 Hz, 1H), 7.51 - 7.44 (m, 2H), 7.39 - 7.30 (m, 3H), 3.96 (s, 2H). 13 C NMR (101 MHz, DMSO-d 6 ) δ 162.28, 155.84, 149.28, 139.35, 134.86, 133.44, 130.78, 129.34, 128.17, 127.43, 127.30, 126.76, 126.19, 121.29, 40.71.

[0063] Synthesis of compound 5 of example 5

[0064]

[0065] Following the synthesis procedure of compound 1, amino terephthalic acid dimethyl ester was replaced by methyl 2-amino-4-methoxybenzoate to give a white solid in 50% yield.

[0066] 1 H NMR (500MHz, DMSO-d) 6 )δ8.02(dd,J=8.9,3.6Hz,1H),7.74(d,J=7.5Hz,1H),7.60(s,1H),7.43–7.3 7(m,1H),7.37–7.32(m,2H),7.18(d,J=8.8Hz,1H),4.25(s,2H),3.90(s,3H). 13 C NMR (101MHz, DMSO-d) 6 )δ164.54,161.81,156.51,151.57,139.37,133.44,130.79,129.33,128.19,127.76,127.30,116.35,114.64,108.54,56.18,56.10,40.01.

[0067] Example 6: Synthesis of Compound 6

[0068]

[0069] Compound 5 (100 mg, 0.33 mmol) was reacted with tetrabutylammonium bromide (106 mg, 0.33 mmol) in 4 mL of hydrobromic acid for 24 hours. The mixture was filtered, and the residue was washed with sodium bicarbonate solution and purified by column chromatography to give 63 mg of white solid, with a yield of 67%.

[0070] 1 H NMR (400MHz, DMSO-d) 6 )δ12.12(s,1H),10.48(s,1H),7.91(d,J=8.7Hz,1H),7.46(s,1H),7.34(dd,J=8 .1, 2.8Hz, 3H), 6.91 (dd, J=8.7, 2.3Hz, 1H), 6.85 (d, J=2.3Hz, 1H), 3.90 (s, 2H). 13 C NMR (126MHz, DMSO-d) 6 )δ164.55,161.82,156.53,139.35,133.43,130.83,129.31,128.19,127.78,127.32,116.42,114.59,108.49,100.00,56.15.

[0071] Example 7 Synthesis of Intermediate M1

[0072]

[0073] Compound 1 (100 mg, 0.3 mmol) was dissolved in a mixture of 3.0 mL of 1 N aqueous lithium hydroxide and 6 mL of ethanol and heated to reflux for 2.5 hours. The reaction was allowed to cool, after which 1 N hydrochloric acid was added to adjust the pH to weakly acidic, and 99 mg of a white solid was precipitated, in 99% yield.

[0074] 1 H NMR (400 MHz, DMSO-d 6 ) δ 13.43 (s, 1H), 12.60 (s, 1H), 8.18 (d, J = 8.2 Hz, 1H), 8.07 (s, 1H), 7.95 (d, J = 9.6 Hz, 1H), 7.49 (s, 1H), 7.36 (t, J = 5.8 Hz, 3H), 3.99 (s, 2H). 13 CNMR (101 MHz, DMSO-d 6 ) δ 166.91, 161.88, 157.00, 148.84, 139.05, 136.55, 133.44, 130.76, 129.40, 128.25, 127.35, 126.84, 126.52, 124.16, 119.67.

[0075] Synthesis of intermediate M2 in Example 8

[0076]

[0077] M1 (500 mg, 1.6 mmol), HATU (1.8 g, 4.8 mmol), ammonium chloride (103 mg, 1.92 mmol) and N,N-diisopropylethylamine (0.84 mL, 3 mmol) were dissolved in 10 mL of N,N-dimethylformamide and reacted at room temperature for 6 hours. The reaction was extracted with water and ethyl acetate, and the organic phase was evaporated under reduced pressure. The solid was washed with ethanol and dried to obtain 480 mg of a white solid in 99% yield.

[0078] 1 H NMR (500 MHz, DMSO-d 6 ) δ 12.49 (d, J = 57.0 Hz, 1H), 8.24 (s, 1H), 8.13 (d, J = 8.1 Hz, 1H), 8.06 (s, 1H), 7.88 (d, J = 8.2 Hz, 1H), 7.58 (s, 1H), 7.46 (s, 1H), 7.35 (dd, J = 10.4, 7.6 Hz, 3H), 3.98 (s, 2H).

[0079] Synthesis of compound 7 in Example 9

[0080]

[0081] In a two-necked flask was added M2 (500 mg, 1.6 mmol), 50 mL of anhydrous dichloromethane and 1.1 mL of triethylamine, stirred, argon protection, slowly drop 0.9 mL of trifluoroacetic anhydride, reaction at room temperature overnight, the solution was rotary evaporated under reduced pressure and then purified by column to obtain 137 mg of white solid, yield 29%.

[0082] 1 H NMR (400 MHz, DMSO-d6) δ 12.75 (s, 1H), 8.21 (d, J = 8.2 Hz, 1H), 8.12 (s, 1H), 7.83 (d, J = 8.2 Hz, 1H), 7.47 (s, 1H), 7.41 - 7.30 (m, 3H), 3.99 (s, 2H). 6 ) δ 12.75 (s, 1H), 8.21 (d, J = 8.2 Hz, 1H), 8.12 (s, 1H), 7.83 (d, J = 8.2 Hz, 1H), 7.47 (s, 1H), 7.41 - 7.30 (m, 3H), 3.99 (s, 2H). 13 CNMR (101 MHz, DMSO-d6) δ 157.89, 151.32, 140.09, 138.89, 135.08, 133.48, 130.79, 129.17, 128.81, 128.24, 127.40, 124.50, 121.03, 118.31, 117.03, 40.70. 6 ) δ 12.75 (s, 1H), 8.21 (d, J = 8.2 Hz, 1H), 8.12 (s, 1H), 7.83 (d, J = 8.2 Hz, 1H), 7.47 (s, 1H), 7.41 - 7.30 (m, 3H), 3.99 (s, 2H).

[0083] Synthesis of compound 8

[0084]

[0085] Referring to the synthesis method of compound 1, dimethyl amino terephthalate was replaced by methyl 2-amino-3-methoxybenzoate to obtain white solid, yield 55%.

[0086] 1 H NMR (500 MHz, DMSO-d6) δ 7.62 (d, J = 7.7 Hz, 1H), 7.45 (s, 1H), 7.37 (dd, J = 15.3, 7.5 Hz, 2H), 7.31 (t, J = 7.3 Hz, 3H), 3.94 (s, 2H), 3.87 (s, 3H). 6 C NMR (126 MHz, DMSO-d6) δ 162.73, 154.98, 154.59, 139.68, 133.42, 130.83, 129.12, 127.98, 127.24, 126.92, 122.21, 117.23, 115.35, 56.33, 41.03. 13 ) δ 12.75 (s, 1H), 8.21 (d, J = 8.2 Hz, 1H), 8.12 (s, 1H), 7.83 (d, J = 8.2 Hz, 1H), 7.47 (s, 1H), 7.41 - 7.30 (m, 3H), 3.99 (s, 2H). 6

[0087] Synthesis of compound 9

[0088] ​

[0089] Referring to the synthesis method of compound 6, compound 5 was replaced by compound 8 to obtain 27 mg of white solid with a yield of 29%.

[0090] 1 H NMR (500 MHz, DMSO-d6) δ 12.38 (s, 1H), 9.54 (s, 1H), 7.54 (s, 1H), 7.50 (d, J = 7.7 Hz, 1H), 7.40 (d, J = 7.2 Hz, 1H), 7.35 (t, J = 7.6 Hz, 1H), 7.31 (d, J = 9.2 Hz, 1H), 7.28 (d, J = 7.8 Hz, 1H), 7.18 (d, J = 7.7 Hz, 1H), 3.95 (s, 2H). 6 C NMR (126 MHz, DMSO-d6) δ 162.33, 154.01, 153.01, 139.51, 138.25, 133.47, 130.80, 129.22, 128.05, 127.33, 127.23, 122.07, 118.78, 115.97. 13 6

[0091] Synthesis of compound 10 of Example 12

[0092]

[0093] Referring to the synthesis method of compound 1, dimethyl amino terephthalate was replaced by methyl 2,3-diaminobenzoate to obtain black solid with a yield of 55%.

[0094] 1 H NMR (400 MHz, DMSO-d6) δ 12.48 (s, 1H), 7.85 (d, J = 7.8 Hz, 1H), 7.78 (d, J = 7.6 Hz, 1H), 7.45 (s, 1H), 7.30 (m, 4H), 4.30 (s, 2H), 3.95 (s, 2H). 6 C NMR (126 MHz, DMSO-d6) δ 166.18, 155.50, 144.77, 140.57, 133.37, 130.73, 129.15, 128.07, 126.94, 124.19, 121.41, 113.88, 52.24. 13 6

[0095] Synthesis of compound 11 of Example 13

[0096] ​​​

[0097] Ml (100 mg, 0.32 mmol) and N,N'-carbonyldiimidazole (100 mg, 0.63 mmol) were dissolved in 3 mL of N,N-dimethylformamide, after stirring for one hour, hydroxylamine hydrochloride (88 mg, 1.27 mmol) was added, stirred for 24 hours, diluted with dilute hydrochloric acid to weakly acidic, the filtrate was removed under reduced pressure, washed with dichloromethane, dried to obtain 27 mg of white solid, yield 13%.

[0098] 1 H NMR (400 MHz, DMSO-d 6 ) δ 12.67 (s, 1H), 11.53 (s, 1H), 9.26 (s, 1H), 8.12 (d, J = 8.2 Hz, 1H), 7.93 (d, J = 1.1 Hz, 1H), 7.81 (dd, J = 8.2, 1.3 Hz, 1H), 7.49 (s, 1H), 7.36 (d, J = 5.4 Hz, 2H), 7.33 (dd, J = 5.7, 3.6 Hz, 1H), 4.02 (s, 2H). 13 C NMR (126 MHz, DMSO-d 6 ) δ 163.47, 161.83, 156.85, 149.06, 139.25, 138.53, 133.38, 130.77, 129.43, 128.31, 127.30, 126.58, 125.76, 124.99, 122.98, 40.52, 39.96.

[0099] Synthesis of compound 12 of example 14

[0100]

[0101] Ml (314 mg, 1 mmol), acethydrazide (78 mg, 1.1 mmol), HATU (450 mg, 1.2 mmol) and N,N-diisopropylethylamine (524 mg, 3 mmol) were dissolved in 5 mL of N,N-dimethylformamide, reacted at room temperature for 30 minutes, extracted with water and ethyl acetate, after the organic phase was removed under reduced pressure, purified by column to obtain 175 mg of white solid, yield 47%.

[0102] 1 H NMR (400 MHz, DMSO-d 6)δ12.58(s,1H),10.53(s,1H),9.96(s,1H),8.17(d,J=8.2Hz,1H),8.07(s,1H),7.88( d,J=9.1Hz,1H),7.49(s,1H),7.36(dd,J=10.3,4.2Hz,3H),4.00(s,2H),1.93(s,3H). 13 C NMR (101MHz, DMSO-d) 6 )δ168.92,165.09,161.88,156.92,149.10,139.12,138.23,133.44,13 0.76,129.48,128.32,127.33,126.73,126.48,125.33,123.40,21.06.

[0103] Example 15 Synthesis of Compound 13

[0104]

[0105] Compound 12 (175 mg, 0.47 mmol) was reacted in 3 mL of phosphorus oxychloride at 80 °C for 5 hours. After the phosphorus oxychloride was evaporated to dryness, the mixture was quenched with ice water. The solution was extracted with ethyl acetate, and the organic phase was purified by rotary evaporation under reduced pressure and column chromatography to give 49 mg of white solid, with a yield of 30%.

[0106] 1 H NMR (400MHz, DMSO-d) 6 )δ12.62(s,1H),8.23(d,J=8.2Hz,1H),8.04(s,1H),7.99(d,J=8.2Hz,1H),7.49(s,1H),7.35(dd,J=12.9,4.6Hz,3H),3.99(s,2H),2.61(s,3H). 13 CNMR (126MHz, DMSO-d) 6 )δ172.03,168.58,164.93,164.57,151.58,142.61,133.04,130.33,129.0 5,128.26,127.67,126.34,126.03,123.71,122.79,120.40,45.53,11.01.

[0107] Example 16 Synthesis of Compound 14

[0108]

[0109] Compound 6 (422 mg, 1.47 mmol), methyl bromoacetate (282 μL, 2.97 mmol) and cesium carbonate (1.44 g, 4.42 mmol) were reacted in 10 mL of acetonitrile at room temperature for 14 hours, and the solution was rotary evaporated under reduced pressure and then purified by column to obtain 73 mg of white solid, with a yield of 14%.

[0110] 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.62 (s, 1H), 7.91 (d, J = 8.8 Hz, 1H), 7.58 (d, J = 39.4 Hz, 2H), 7.44 (t, J = 7.9 Hz, 1H), 7.28 (d, J = 7.9 Hz, 1H), 6.99 (d, J = 2.3 Hz, 1H), 6.88 (dd, J = 8.8, 2.4 Hz, 1H), 4.92 (s, 2H), 4.19 (s, 2H), 3.72 (s, 3H). 13 C NMR (126 MHz, DMSO-d 6 ) δ 169.04, 162.94, 158.68, 156.18, 133.42, 133.00, 130.51, 129.02, 128.25, 127.14, 126.15, 112.30, 110.26, 65.24, 52.54, 52.47, 51.46.

[0111] Synthesis of compound 15 of example 17

[0112]

[0113] Referring to the synthesis method of intermediate M1, compound 1 was replaced by compound 14 to obtain a white solid, with a yield of 99%.

[0114] 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.66 (s, 1H), 7.90 (d, J = 8.8 Hz, 1H), 7.55 (d, J = 36.1 Hz, 2H), 7.44 (t, J = 7.8 Hz, 1H), 7.29 (d, J = 7.7 Hz, 1H), 6.99 (d, J = 1.7 Hz, 1H), 6.86 (dd, J = 8.8, 2.1 Hz, 1H), 4.79 (s, 2H), 4.19 (s, 2H). 13 C NMR (126 MHz, DMSO-d 6) δ 169.98, 163.16, 158.82, 156.28, 133.36, 133.13, 130.46, 128.88, 128.10, 127.04, 126.01, 112.59, 110.05, 101.52, 96.84, 65.17, 51.46.

[0115] Synthesis of intermediate M3 in example 18

[0116]

[0117] The synthesis of reference compound 1 was followed, replacing dimethyl amino terephthalate with methyl 2-amino-4-bromobenzoate, to give a white solid in 50% yield.

[0118] 1H NMR (400 MHz, DMSO-d6) δ 7.99 (d, J = 8.4 Hz, 1H), 7.88 (s, 1H), 7.67 (d, J = 8.5 Hz, 1H), 7.53 (s, 1H), 7.35 - 7.29 (m, 3H), 4.04 (s, 2H). 6 ) δ 169.98, 163.16, 158.82, 156.28, 133.36, 133.13, 130.46, 128.88, 128.10, 127.04, 126.01, 112.59, 110.05, 101.52, 96.84, 65.17, 51.46.

[0119] Synthesis of intermediate M4 in example 19

[0120]

[0121] M3 (100 mg, 0.29 mmol), 135 μL of triethylamine, methyl acrylate (130 μL, 0.45 mmol), triphenylphosphine (20 mg, 0.26 mmol) and palladium acetate (9 mg, 0.13 mmol) were added in N,N-dimethylformamide and left to react overnight under argon protection. The reaction was extracted with water and ethyl acetate and the organic phase was evaporated under reduced pressure and purified on a column to give 72 mg of a white solid in 60% yield.

[0122] 1 H NMR (400 MHz, DMSO-d6) δ 12.48 (s, 1H), 8.07 (d, J = 8.2 Hz, 1H), 7.91 (s, 1H), 7.83 (d, J = 8.3 Hz, 1H), 7.77 (d, J = 16.1 Hz, 1H), 7.47 (s, 1H), 7.34 (q, J = 7.2 Hz, 3H), 6.84 (d, J = 16.1 Hz, 1H), 3.96 (s, 2H), 3.74 (s, 3H). 6 ) δ 169.98, 163.16, 158.82, 156.28, 133.36, 133.13, 130.46, 128.88, 128.10, 127.04, 126.01, 112.59, 110.05, 101.52, 96.84, 65.17, 51.46.

[0123] Synthesis of compound 16 in example 20

[0124]

[0125] Referring to the synthesis method of intermediate Ml, the compound 1 was replaced by M4 to obtain a white solid with a yield of 99%.

[0126] 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.07 (d, J = 8.3 Hz, 1H), 7.87 (s, 1H), 7.81 (d, J = 8.3 Hz, 1H), 7.69 (d, J = 16.0 Hz, 1H), 7.48 (s, 1H), 7.35 (dd, J = 9.7, 4.4 Hz, 3H), 6.72 (d, J = 16.1 Hz, 1H), 3.98 (s, 2H). 13 C NMR (126 MHz, DMSO-D6) δ 171.14, 164.95, 149.96, 142.07, 140.09, 136.05, 133.34, 131.79, 130.72, 129.13, 128.15, 127.09, 126.62, 125.25, 124.23, 120.46, 40.36.

[0127] Synthesis of intermediate M5 in Example 21

[0128]

[0129] Potassium tert-butoxide (259 mg, 2.3 mmol) was added to 2 mL of N,N- dimethylformamide, stirred at -15 °C, argon protection, slowly added 4-nitrochlorobenzene (158 mg, 1 mmol) and 4-chlorophenoxyacetonitrile (202 mg, 1.2 mmol) in N,N- dimethylformamide 100 mL, reacted overnight, extracted with water and ethyl acetate, the organic phase was rotary evaporated under reduced pressure and then purified by column to obtain 45 mg of white solid with a yield of 23%.

[0130] 1 H NMR (500 MHz, DMSO-d 6 ) δ 8.20 (d, J = 8.8 Hz, 1H), 7.84 (s, 1H), 7.76 (dd, J = 8.8, 1.9 Hz, 1H), 4.34 (s, 2H).

[0131] Synthesis of intermediate M6 in Example 22

[0132]

[0133] Referring to the synthesis method of compound 1, 3-chlorophenylacetonitrile was replaced by M5, and dimethyl amino terephthalate was replaced by methyl 2-amino-4- bromobenzoate to obtain a white solid with a yield of 72%.

[0134] 1 H NMR (400MHz, DMSO-d) 6 )δ8.16(d,J=8.8Hz,1H),7.99(d,J=8.5Hz,1H),7.80(d,J=2.2Hz,1H),7.70(dd,J =8.8,2.3Hz,1H),7.63(dd,J=8.5,1.8Hz,1H),7.54(d,J=1.7Hz,1H),4.43(s,2H).

[0135] Example 23 Synthesis of intermediate M7

[0136]

[0137] M6 (393 mg, 1 mmol) and anhydrous stannous chloride (1 g, 5.3 mmol) were added to 5 mL of concentrated hydrochloric acid and reacted overnight at 60 °C. The mixture was filtered, and the residue was washed with concentrated hydrochloric acid and ethyl acetate and dried to give a white solid with a yield of 55%.

[0138] 1 H NMR (400MHz, DMSO-d) 6 )δ8.00(d,J=8.5Hz,1H),7.87(d,J=1.7Hz,1H),7.66(dd,J=8.5,1.9Hz,1H),7.53(s,1H),7.50–7.43(m,2H),4.22(s,2H).

[0139] Example 24 Synthesis of intermediate M8

[0140]

[0141] Following the synthesis method of intermediate M4, M3 was replaced with M7, yielding 200 mg of white solid with a yield of 33%.

[0142] 1 H NMR (400MHz, DMSO-d) 6 )δ12.43(s,1H),8.07(d,J=8.2Hz,1H),7.92(s,1H),7.83(d,J=8.3Hz,1H),7.78(d,J=16.1Hz,1H),7.18(d,J=2.4Hz,1 H),7.00(dd,J=8.5,2.3Hz,1H),6.85(d,J=16.1Hz,1H),6.68(d,J=8.6Hz,1H),5.40(s,2H),3.78(s,2H),3.75(s,3H).

[0143] Synthesis of intermediate M9 in example 25

[0144]

[0145] Referring to the synthesis method of intermediate M4, replace M3 with M7 and methyl acrylate with tert-butyl acrylate to obtain a white solid with a yield of 35%.

[0146] 1 H NMR (400 MHz, DMSO-d6) δ 12.42 (s, 1H), 8.06 (d, J = 8.3 Hz, 1H), 7.90 - 7.75 (m, 3H), 7.66 (d, J = 16.1 Hz, 1H), 7.30 (s, 1H), 7.22 (s, 1H), 6.71 (d, J = 16.0 Hz, 1H), 5.60 (s, 2H), 3.89 (s, 2H), 1.50 (s, 9H). 6

[0147] Synthesis of intermediate M10 in example 26

[0148]

[0149] M9 (65 mg, 0.24 mmol), acetyl chloride (57 μL, 0.37 mmol) and triethylamine (67 μL, 0.48 mmol) were added to 50 mL of N,N-dimethylformamide and reacted at room temperature overnight, extracted with water and ethyl acetate, and the organic phase was rotary evaporated under reduced pressure and then purified by column to obtain a white solid with a yield of 32%.

[0150] 1 H NMR (400 MHz, DMSO-d6) δ 12.35 (s, 1H), 8.06 (d, J = 8.6 Hz, 1H), 7.82 (dd, J = 10.3, 6.0 Hz, 2H), 7.70 - 7.63 (m, 1H), 7.58 (d, J = 8.6 Hz, 1H), 7.46 (d, J = 2.2 Hz, 1H), 7.33 (dd, J = 8.9, 1.9 Hz, 1H), 6.71 (dd, J = 16.0, 3.8 Hz, 1H), 3.97 (d, J = 8.3 Hz, 2H), 2.13 (s, 2H), 1.50 (s, 9H). 6

[0151] Synthesis of intermediate M11 in example 27

[0152]

[0153] Referring to the synthesis method of intermediate M10, replace acetyl chloride with cyclohexanecarbonyl chloride to obtain a white solid with a yield of 35%. ​​

[0154] 1 H NMR (400MHz, DMSO-d) 6 )δ12.31(s,1H),8.06(dd,J=8.2,5.2Hz,1H),7.81(t,J=8.5Hz,1H),7.76(d,J=6 .3Hz,1H),7.72–7.58(m,3H),7.47(d,J=2.3Hz,1H),6.68(dd,J=16.1,4.3Hz,1H) ,3.95(d,J=12.6Hz,2H),2.33(t,J=11.7Hz,1H),1.87–1.78(m,2H),1.69(s,2H), 1.65–1.57(m,1H),1.49(s,9H),1.39–1.32(m,2H),1.20(dd,J=16.3,6.9Hz,3H).

[0155] Example 28 Synthesis of intermediate M12

[0156]

[0157] Following the synthesis method of intermediate M10, M9 was replaced with M8, and acetyl chloride was replaced with 3-(trifluoromethoxy)benzoyl chloride, yielding a white solid with a yield of 32%.

[0158] 1 H NMR (400MHz, DMSO-d) 6 )δ12.45(s,1H),10.60(s,1H),8.04(d,J=8.7Hz,2H),7.90(s,1H),7.80(d,J=7.5Hz,1H),7.69(dd,J=15.7,7.7Hz,3H),7.62 (d,J=10.8Hz,1H),7.58(s,1H),7.50(s,1H),7.42(dd,J=9.8,1.8Hz,1H),6.69(d,J=15.3Hz,1H),4.09(s,2H),3.75(s,3H).

[0159] Example 29 Synthesis of intermediate M13

[0160]

[0161] M8 (100 mg, 0.27 mmol), 3-fluorobenzaldehyde (135 mg, 1.1 mmol) and sodium triacetoxyborohydride (572 mg, 2.7 mmol) were reacted overnight at room temperature in anhydrous 1,2-dichloroethane. The reaction solution was purified by rotary evaporation under reduced pressure and column chromatography to give 57 mg of white solid, yield 45%.

[0162] 1 H NMR (400MHz, DMSO-d) 6 )δ12.51(s,1H),8.09(d,J=8.3Hz,1H),7.84(dd,J=8.4,1.3Hz,1H),7.78–7.71 (m,2H),7.31(dd,J=13.9,7.8Hz,1H),7.26(d,J=2.5Hz,1H),7.18(dd,J=20.3, 8.8Hz,2H),7.05(dd,J=8.7,2.6Hz,2H),6.80(d,J=16.1Hz,1H),6.50(t,J=5.1 Hz,1H),6.46(d,J=8.8Hz,1H),4.40(d,J=5.7Hz,2H),3.92(s,2H),3.77(s,3H).

[0163] Example 30 Synthesis of intermediate M14

[0164]

[0165] Following the synthesis method of intermediate M13, 3-fluorobenzaldehyde was replaced with benzaldehyde to obtain a white solid with a yield of 40%.

[0166] 1 H NMR (400MHz, DMSO-d) 6 )δ12.52(s,1H),8.08(d,J=7.3Hz,1H),7.83(d,J=7.0Hz,1H),7.73(d,J=17.4Hz,1H),7.63(s,1H),7.41–7.34(m,2H),7.33 –7.20(m,4H),7.05(d,J=7.3Hz,1H),6.79(d,J=17.0Hz,1H),6.51(d,J=3.9Hz,1H),4.35(s,2H),3.89(s,2H),3.77(s,3H).

[0167] Example 31 Synthesis of intermediate M15

[0168]

[0169] Following the synthesis method of intermediate M13, 3-fluorobenzaldehyde was replaced with 2-benzaldehyde to obtain a white solid with a yield of 20%.

[0170] 1 H NMR (400MHz, DMSO-d) 6)δ12.51(s,1H),8.07(d,J=7.3Hz,1H),7.83(d,J=8.2Hz,1H),7.77–7.67(m,2H),7.38(t,J=7.6Hz,1H),7.33–7.24(m,2H),7.22–7.1 4(m,1H),7.09–7.04(m,1H),6.80(d,J=15.5Hz,1H),6.51(d,J=8.3Hz,1H),6.43–6.35(m,1H),4.40(s,2H),3.90(s,2H),3.77(s,3H).

[0171] Example 32 Synthesis of intermediate M16

[0172]

[0173] Following the synthesis method of intermediate M13, 3-fluorobenzaldehyde was replaced with 4-benzaldehyde to obtain a white solid with a yield of 23%.

[0174] 1 H NMR (400MHz, DMSO-d) 6 )δ12.50(s,1H),8.08(d,J=8.2Hz,1H),7.84(d,J=8.2Hz,1H),7.69(s,1H),7.62(d,J=6.6Hz,2H),7.41–7.36(m,2H),7.25(s ,1H),7.10(s,1H),6.81(d,J=16.2Hz,1H),6.49(d,J=8.7Hz,1H),6.46–6.42(m,1H),4.34(s,2H),3.90(s,2H),3.76(s,3H).

[0175] Example 33 Synthesis of intermediate M17

[0176]

[0177] Following the synthesis method of intermediate M13, 3-fluorobenzaldehyde was replaced with 3'-fluoroacetophenone to obtain a white solid with a yield of 25%.

[0178] 1 H NMR (400MHz, DMSO-d) 6)δ12.51(s,1H),8.07(d,J=8.7Hz,1H),7.89(s,1H),7.84(d,J=6.8Hz,1H),7.62(d,J=6.6Hz,2H),7.51–7.49(m,2H),7.29(d,J=9.0 Hz,2H),7.12(d,J=10.0Hz,1H),6.84(d,J=16.6Hz,1H),6.59(d,J=8.9Hz,1H),3.88(s,1H),3.81(s,2H),3.75(s,3H),1.24(m,3H).

[0179] Example 34 Synthesis of intermediate M18

[0180]

[0181] Following the synthesis method of intermediate M13, 3-fluorobenzaldehyde was replaced with benzo[d][1,3]dioxane-5-carboxaldehyde to obtain a white solid with a yield of 25%.

[0182] 1 H NMR (500MHz, DMSO-d) 6 )δ12.44(s,1H),8.14(dd,J=8.2,2.1Hz,1H),8.01(d,J=17.8Hz,1H),7.93( d,J=9.1Hz,1H),7.86(d,J=13.0Hz,1H),7.82–7.73(m,1H),7.52(d,J=8.6H z,1H),7.26(d,J=8.4Hz,2H),7.15(d,J=8.1Hz,1H),7.04(d,J=8.2Hz,1H), 6.87(d,J=16.1Hz,1H),6.11(s,2H),4.22(s,2H),3.76(s,2H),3.59(s,3H).

[0183] Example 35 Synthesis of Compound 17

[0184]

[0185] M10 (40 mg, 0.08 mmol) was added to dichloromethane and 0.4 mL of trifluoroacetic acid was slowly added dropwise under ice bath. The reaction was allowed to proceed overnight. The reaction solution was purified by rotary evaporation under reduced pressure and column chromatography to obtain a white solid with a yield of 87%.

[0186] 1 H NMR (400MHz, DMSO-d) 6)δ9.99(s,1H),9.67(s,1H),8.05(d,J=8.0Hz,1H),7.74(d,J=9.9Hz,2H),7.64–7.54(m,2H),7.48(dd,J =18.4,1.9Hz,1H),7.32(dd,J=8.9,2.2Hz,1H),6.68(dd,J=15.9,2.5Hz,1H),3.97(s,2H),2.01(s,3H). 13 C NMR (126MHz, DMSO-d) 6 )δ169.22,161.85,155.96,149.16,141.26,141.03,138.89,136.41,134.21,133.7 5,131.81,130.42,130.04,128.16,127.67,126.92,125.17,121.60,40.54,22.98.

[0187] Example 36 Synthesis of Compound 18

[0188]

[0189] Following the synthetic method of compound 17, replacing M10 with M11 yielded a white solid with a yield of 99%.

[0190] 1 H NMR (400MHz, DMSO-d) 6 )δ9.82(s,1H),9.45(s,1H),8.05(dd,J=8.2,3.5Hz,1H),7.76(t,J=7.7Hz,1H),7.70(s,1H),7.60(m,2H),7.46(s,1H),7.31(dd,J=8.7,2.6Hz, 1H), 6.67 (dd, J=15.9, 2.9Hz, 1H), 3.96 (d, J=12.7Hz, 2H), 2.32 (t, J=11.2Hz, 1H), 1.80 (dd, J=33.5, 19.0Hz, 4H), 1.28 (dd, J=53.6, 11.4Hz, 6H). 13 C NMR (101MHz, DMSO-d) 6)δ174.93,174.76,161.88,155.91,140.74,138.75,136.52,134.24,133.81,131.72,130.53, 130.07,128.17,127.73,126.86,126.69,125.16,121.88,44.20,29.62,29.47,25.91,25.66.

[0191] Example 37 Synthesis of Compound 19

[0192]

[0193] Following the synthetic method of intermediate M1, compound 1 was replaced with M12, yielding a white solid with a yield of 71%.

[0194] 1 H NMR (500MHz, DMSO-d) 6 )δ10.66(s,1H),8.05(dd,J=11.7,8.1Hz,2H),7.91(s,1H),7.76(d,J=7.3Hz,1H),7.69(s,2H ),7.65–7.53(m,3H),7.46(s,1H),7.42(d,J=8.2Hz,1H),6.57(d,J=15.9Hz,1H),4.09(s,2H). 13 C NMR (126MHz, DMSO-d) 6 )δ167.76,164.43,161.84,156.79,148.99,148.82,142.35,140.49,137.11,136.17,133.06,131.14,13 0.99,130.07,128.00,127.92,127.32,126.79,126.51,125.56,124.60,123.34,121.91,120.72,37.71.

[0195] Example 38 Synthesis of Compound 20

[0196]

[0197] Following the synthetic method of intermediate M1, compound 1 was replaced with M13, yielding a white solid with a yield of 67%.

[0198] 1 H NMR (400MHz, DMSO-d) 6)δ12.62(s,1H),8.06(d,J=8.2Hz,1H),7.77(d,J=7.4Hz,1H),7.65(d,J=15.9Hz,1H),7.56(s,1H),7.37(d,J=6.8Hz,2H),7.27 (dd,J=12.2,4.9Hz,4H),7.04(dd,J=8.7,2.4Hz,1H),6.66(d,J=16.1Hz,1H),6.50(d,J=8.8Hz,1H),4.34(s,2H),3.90(s,2H). 13 C NMR (126MHz, DMSO-d) 6 )δ167.66,161.85,156.38,148.86,146.16,142.80,140.40,139.91,130.38,128.75,128.09,1 27.82,127.28,126.86,126.81,125.64,122.90,122.65,122.00,119.93,112.52,47.04,37.71.

[0199] Example 39 Synthesis of Compound 21

[0200]

[0201] Following the synthetic method of intermediate M1, compound 1 was replaced with M14, yielding a white solid with a yield of 75%.

[0202] 1 H NMR (400MHz, DMSO-d) 6 )δ12.56(s,1H),8.06(d,J=8.2Hz,1H),7.78(d,J=7.7Hz,1H),7.69–7.61(m,2H),7.38(t,J=7.2Hz,1H),7.29(d,J=2.4 Hz,2H),7.21–7.12(m,1H),7.11–7.01(m,2H),6.67(d,J=16.1Hz,1H),6.50(d,J=8.8Hz,1H),4.39(s,2H),3.91(s,2H). 13 C NMR (126MHz, DMSO-d) 6)δ167.65,161.80,159.61,156.42,149.01,145.85,142.84,140.41,130.47,129.91,129.86,129.26,12 8.16,127.00,126.75,125.59,124.68,122.91,122.87,122.03,120.12,115.70,115.48,112.20,37.59.

[0203] Example 40 Synthesis of Compound 22

[0204]

[0205] Following the synthetic method of intermediate M1, compound 1 was replaced with M15, yielding a white solid with a yield of 52%.

[0206] 1 H NMR (400MHz, DMSO-d) 6 )δ7.98(d,J=8.1Hz,1H),7.56(d,J=8.2Hz,1H),7.44(s,1H),7.33–7.14(m,5H),7. 02(m,2H),6.53(d,J=15.9Hz,1H),6.43(d,J=8.7Hz,1H),4.38(s,2H),3.88(s,2H). 13 C NMR (126MHz, DMSO-d) 6 )δ170.27,163.81,161.87,149.32,145.90,143.36(d,J=6.7Hz),142.75,135.62,132.84,130.61(d,J=8.3Hz),130. 47,127.92,126.58,125.37,124.63,123.61,123.36,120.71,119.97,114.25,114.07,113.88,112.41,46.33,37.91.

[0207] Example 41 Synthesis of Compound 23

[0208]

[0209] Following the synthetic method of intermediate M1, compound 1 was replaced with M16, yielding a white solid with a yield of 57%.

[0210] 1 H NMR (400MHz, DMSO-d) 6)δ12.50(s,1H),8.07(d,J=8.2Hz,1H),7.81(d,J=8.3Hz,1H),7.71–7.61(m,2H),7.40(dd,J=8.2,5.8Hz,2H),7. 26(d,J=2.4Hz,1H),7.14–7.00(m,3H),6.69(d,J=16.1Hz,1H),6.49(d,J=8.8Hz,1H),4.35(s,2H),3.90(s,2H). 13 CNMR (126MHz, DMSO-d) 6 )δ167.66(s),161.86(s),156.27(s),149.06(s),145.99(s),142.87( s),140.48(s),136.08(s),130.37(s),129.53(d,J=7.7Hz),128.07(s) ,127.14(s),126.78(s),125.60(s),122.92(s),122.72(s),122.01(s) ),119.95(s),115.53(s),115.36(s),112.50(s),46.14(s),37.69(s).

[0211] Example 42 Synthesis of Compound 24

[0212]

[0213] Following the synthetic method of intermediate M1, compound 1 was replaced with M17, yielding a white solid with a yield of 42%.

[0214] 1 H NMR (400MHz, DMSO-d) 6 )δ8.06(d,J=8.2Hz,2H),7.85(d,J=7.4Hz,1H),7.82–7.77(m,2H),7.69(d,J=15.2Hz,2H),7. 27(d,J=12.6Hz,2H),7.11–7.05(m,1H),6.73(s,1H),6.69(s,1H),3.88(s,2H),3.84(s,2H). 13 C NMR (126MHz, DMSO-d) 6)δ167.21,161.42,155.82,152.89,148.62,145.54,142.42,140.03,135.64,129.92,129.11,129.05,127. 62,126.69,126.33,125.15,122.48,122.27,121.59,119.50,115.08,114.91,112.06,45.69,40.11,37.24.

[0215] Example 43 Synthesis of Compound 25

[0216]

[0217] Following the synthetic method of intermediate M1, compound 1 was replaced with M18, yielding a white solid with a yield of 46%.

[0218] 1 H NMR (500MHz, DMSO-d) 6 )δ12.44(s,1H),8.14(dd,J=8.2,2.1Hz,1H),8.02(s,1H),7.96–7.89(m,1H),7.88(d,J=10.1Hz,1H),7.83(d,J=16.2Hz,1H),7.52(d,J=8.6 Hz,1H),7.26(d,J=8.4Hz,2H),7.15(d,J=8.1Hz,1H),7.04(d,J=8.2Hz,1H),6.87(d,J=16.1Hz,1H),6.11(s,2H),4.23(s,2H),3.76(s,2H). 13 CNMR (126MHz, DMSO-d) 6 )δ167.63,166.26,161.44,148.68,148.12,143.24,140.62,134.69,128.85,127.15,127.11,125.96,1 25.86,124.50,123.82,123.35,123.32,122.21,121.78,119.48,113.87,109.28,102.08,60.84,14.64.

[0219] Application Example 1: Inhibitory activity of compound against PDE8A enzyme

[0220] Determination of the required enzyme concentration: A series of gradient concentrations of enzyme solution were prepared with Assay Buffer (1.0 mM magnesium chloride, 50 mM Tris 8.0, 1.0 mM DTT). In each reaction tube, 55.5 μL Assay Buffer, 2.5 μL diluted 3 H-cGMP and 2 μL DMSO-D6 were added. The blank group was added with 40 μL Assay Buffer in the reaction tube, and the experimental group was added with 40 μL protein solution of different concentrations at intervals of 20 seconds. After reaction at room temperature for 15 minutes, 200 μL ZnSO4 (0.2 M) and 200 μL Ba(OH)2 (0.2 M) were added in turn and vortexed, and the reaction tube was centrifuged in a high-speed centrifuge (14000 rpm, 6 min). The corresponding amount of scintillation tube was added with 2.5 mL scintillation solution for standby. After centrifugation, 430 μL supernatant was taken and added to the scintillation tube and vortexed, and then read in a liquid scintillation instrument. The protein concentration between the hydrolysis rate of 50-70% was selected for the subsequent test of the inhibitory activity of the compound.

[0221] Determination of the inhibitory activity of the test compound on the enzyme: The blank group, the control group, the positive group and the compound group were set, and 55.5 μL Assay Buffer and 2.5 μL diluted 3 H-cGMP were added in each reaction tube. The blank group and the control group were added with 2 μL DMSO-D6, the positive group was added with 2 μL sildenafil (100 nM), and the compound group was added with 2 μL DMSO-D6 solution of the test compound. The blank group was added with 40 μL Assay Buffer, and the other three groups were added with 40 μL protein solution of the test concentration at intervals of 20 seconds. The subsequent operation was the same as above. For the compound whose IC 50 needs to be measured, 7-9 gradient concentrations of the test compound need to be taken, and the test operation needs to be repeated 2-3 times. The results are shown in Table 1.

[0222] Table 1 Inhibitory activity of the compound on PDE5

[0223]

[0224] As can be seen from the table, the compounds prepared in the application have different degrees of inhibition on PDE5A. Application Example 2 Selectivity of the compound on other PDE subtypes

[0225] Compound 22 was taken as a representative compound, and its selectivity on other PDE subtypes was tested. The results of other compounds were similar to those of compound 22, and the test results are shown in Table 2.

[0226] Table 2 Selectivity of compound 22 on other PDE subtypes

[0227]

[0228]

[0229] As can be seen from the table, the compound of the application has good inhibitory activity on phosphodiesterase type 5, and lower inhibitory activity on other subtypes of the family, and has a broad application space as a phosphodiesterase type 5 inhibitor.

[0230] Application Example 3 Compound 22 Drugability Evaluation

[0231] The drugability of compound 22 was evaluated, and the acute toxicity test method was as follows: on the first day, the model group of mice was given a single oral dose of 1500 mg / kg of compound 22, and the control group of mice was given an equal amount of normal saline, and the mice were observed for behavior and the body weight of the mice was recorded within the next 14 days. The results showed that: the control group and the model group did not show any symptoms of poisoning or death, and no abnormal behavior or significant changes in water / food consumption and body weight were observed during the experiment. Therefore, the mice had good tolerance when the dose was as high as 1500 mg / kg, and it was determined that the acute toxicity of compound 22 was greater than 1.5 g / kg.

[0232] The remaining parameters were detected by the Chinese Academy of Sciences Shanghai Institute of Materia Medica (the detection method is referred to in [1] Kutchinsky J, Friis S, Asmild M. et al (2003) Characterization of potassium channel modulators with QPatch automated patch-clamp technology: system characteristics and performance, Assay Drug Dev Technol 1(5): 685-693. [2] Zhou PZ, Babcock J, Liu LQ, Li M, Gao ZB (2011) Activation of human ether-a-go-go related gene (hERG) potassium channels by small molecules, Acta Pharmacol Sin 32(6): 781-8. [3] Zhou Z, Gong Q, Ye B. et al (1998) Properties of HERG channels stably expressed in HEK 293 cells studied at physiological temperature. Biophysical Journal; 74: 230-241.), and the results are shown in Table 2.Figure 1 and Table 3.

[0233] Table 3 Drug-likeness evaluation of compound 22

[0234]

[0235]

[0236] The results show that the IC50 of compound 22 for hERG inhibition is greater than 40 μM, indicating that the compound does not cause cardiac toxicity of arrhythmia. The acute toxicity experiment in mice shows that the tolerance dose of compound 22 is as high as 1.5 g / kg, indicating that compound 22 is safe at a dose of 1.5 g / kg. 50 The results show that the IC50 of compound 22 for hERG inhibition is greater than 40 μM, indicating that the compound does not cause cardiac toxicity of arrhythmia. The acute toxicity experiment in mice shows that the tolerance dose of compound 22 is as high as 1.5 g / kg, indicating that compound 22 is safe at a dose of 1.5 g / kg.

[0237] Example 4 Pharmacodynamic study of compound 22 against pulmonary fibrosis

[0238] Grouping of pulmonary fibrosis model rats: The rats were kept in a 12-hour light-dark cycle at a relative humidity of 60-70% and a temperature of 24±1℃. After a one-week adaptation period, the rats were randomly divided into four groups (15 rats per group): a control group, a model group, a compound 22 (2.5 mg / kg) group, and a positive control group (pirfenidone, PFD, 150 mg / kg).

[0239] Modeling of pulmonary fibrosis model rats: The control group was subjected to sham operation with normal saline, and all other rats were modeled by intratracheal single instillation of bleomycin (5 mg / kg).

[0240] Administration of pulmonary fibrosis model rats: The control and model groups were intraperitoneally injected with normal saline, the positive group was administered pirfenidone by gavage, and the experimental group was intraperitoneally injected with compound 22, once a day, for four weeks.

[0241] Observation and recording: The body weight was observed and recorded every three days. After four weeks of administration, the respiratory level of each group was measured, including: respiratory rate, respiratory interval, peak expiratory flow, peak inspiratory flow, and airway resistance parameters.

[0242] Sampling: The rats were anesthetized by intraperitoneal injection of 4% sodium pentobarbital, 5 mL of blood was drawn from the abdominal aorta, and after centrifugation, it was stored in a refrigerator at -80℃. Subsequently, the rats were euthanized and the left lower lung tissue was taken.

[0243] Preparation and experimental detection of lung tissue: The tissue was soaked in 4% paraformaldehyde overnight, then embedded with paraffin, and then cut into sections with a thickness of 5 μm. The sections were fixed on glass slides, and the tissue sections were stained with hematoxylin-eosin (H&E) or Masson, observed under an optical microscope, and photographed.

[0244] The enhanced pause (penH), the effective respiratory number (n) and the like are important indexes reflecting airway resistance and evaluating respiratory dysfunction. The detection results of the respiratory indexes of the rats are shown in Table 1. Figure 2 As can be seen, the indexes penH and n of the model group rats obviously rise, indicating that the BLM-induced IPF rat model is successfully constructed. The indexes penH and n of the positive control pirfenidone (PFD) group at a dose of 150 mg / kg obviously decrease compared with the model group, while the indexes penH and n of the compound 22 administration group at a dose of 2.5 mg / kg obviously decrease, indicating that the compound 22 can significantly improve the respiratory function of the rats.

[0245] The H&E staining results are shown in Figure 3. Figure 3 As can be seen from the figure, compared with the blank group, the lung tissue of the model group rats has inflammatory cell infiltration, epithelial cell degeneration, disordered alveolar arrangement and ruptured and fused alveolar wall; the compound 22 administration group at a dose of 2.5 mg / kg significantly reduces the pulmonary fibrosis and the lung damage is lighter.

[0246] The Masson staining and the collagen area ratio of the section are shown in Figure 4. Figure 4 As can be seen from the figure, compared with the blank group, the lung tissue of the model group rats has a large amount of collagen deposition and cell proliferation around the trachea; the compound 22 administration group at a dose of 2.5 mg / kg can inhibit the collagen deposition to the near normal level.

[0247] The Western blotting and the result statistics of the expression of α-smooth muscle actin (α-SMA) in the lung tissue of the pulmonary fibrosis rats are shown in Figure 5. Figure 5 As can be seen from the figure, compared with the blank group, the α-SMA level of the lung of the model group rats is significantly up-regulated, and the compound 22 can effectively reduce the α-SMA level.

[0248] The above examples are the preferred embodiments of the present application, but the embodiments of the present application are not limited by the above examples, and any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present application shall be equivalent replacement modes and shall be included in the protection scope of the present application.

Claims

1. A dihydroquinolinone derivative, characterized by, The dihydroquinolinone derivative has any one of the following structures: 。 2. The dihydroquinolinone derivative according to claim 1, wherein The dihydroquinolinone derivative also includes pharmaceutically acceptable salts thereof.

3. A process for the preparation of a dihydroquinolinone derivative according to claim 1 or 2, characterized in that, When the dihydroquinolinone derivative has any one of the following structures: The synthetic route is as follows: Specifically comprising the steps of: i. Compound A is reacted with compound B in the presence of a base to form compound C completely; ii. Compound C is reacted with compound D in the presence of an acid to form compound E completely; iii. Compound E is reacted with a reducing agent to form compound F; iv. Compound F is reacted with N,N - dimethylformamide, a base and an acrylate ester in the presence of a catalyst to form Compound G; v. Compound G is reacted with compound H in the presence of a reducing agent to form compound I completely; vi. Compound I is reacted with a base to form compound J completely; wherein R 10 is hydrogen, halogen, hydroxy or methyl; X is -CO-Ra, -CH2-Ra or -CH(CH3)-Ra; R 1~4 , R6, R8, R9, Ra are defined as in claim 1.

4. The preparation method according to claim 3, characterized in that, In steps i, iv and vi, the base is selected from one or more of diisopropylethylamine, triethylamine, 4-dimethylaminopyridine, piperidine, sodium bicarbonate, sodium carbonate, potassium carbonate, sodium hydroxide, potassium hydroxide, potassium tert-butoxide, sodium hydride, sodium methoxide, sodium ethoxide.

5. The preparation method according to claim 3, characterized in that, In step ii, the acid is selected from one or more of hydrochloric acid, sulfuric acid, hydrogen chloride, 4M hydrogen chloride in dioxane, 4M hydrogen halide in methanol.

6. Use of a dihydroquinolinone derivative for the preparation of a phosphodiesterase type V inhibitor, characterized in that, The dihydroquinolinone derivative has any one of the following structures: 。 7. Use of a dihydroquinolinone derivative for the preparation of a medicament for the treatment of a disease associated with phosphodiesterase type 5, characterized in that, The dihydroquinolinone derivative has any one of the following structures: 。 8. Use according to claim 7, characterized in that, The phosphodiesterase type 5 related disease is selected from erectile dysfunction, pulmonary arterial hypertension, organ fibrosis, female sexual dysfunction, premature labor, dysmenorrhea, benign prostatic hyperplasia, tumor multidrug resistance, bladder outlet obstruction, incontinence, unstable and variant angina, hypertension, congestive heart failure, renal failure, atherosclerosis, stroke, peripheral vascular disease, Raynaud's disease, inflammatory disease, chronic asthma, allergic asthma, allergic rhinitis, glaucoma, diseases associated with intestinal motility disorders.

Images