A diaryl unsaturated ketone compound, a preparation method thereof, an ALK4 inhibitor and application thereof

Abstract

The application provides a diaryl unsaturated ketone compound and a preparation method, ALK4 inhibitor and application thereof, a chemical structural formula of which is shown in the following; the diaryl saturated or unsaturated ketone compound is used for preparing the ALK4 inhibitor. The diaryl saturated ketone compound provided by the application has multiple R1-R2 substituents, and the compound with the novel structure can fill the gap in the development of systematic ALK4 inhibitors. The ALK4 inhibitor of the diaryl saturated or unsaturated ketone compound has excellent inhibition effect, and test results show that the compound after specific modification can specifically target ALK4, and has significant advantages in disease treatment.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to a diaryl unsaturated ketone compound, its preparation method, an ALK4 inhibitor, and its applications. Background Technology

[0002] ALK4 (Activin Receptor-Like Kinase 4) is a key receptor kinase in the TGF-β superfamily signaling pathway, primarily involved in Activin / Nodal signal transduction and regulating cell proliferation, differentiation, apoptosis, and embryonic development. ALK4 inhibitors, by selectively blocking the kinase activity of ALK4, have potential therapeutic value in cancer, fibrotic diseases, and metabolic disorders. Current ALK inhibitors mainly face two major challenges: drug resistance (especially "compound mutations") and treatment safety (off-target effects). Their specific iterative progress and problems are as follows:

[0003] To address the aforementioned shortcomings, the design and synthesis of ALK4 inhibitors have significant research value in the field of biomedicine. Summary of the Invention

[0004] The technical problem this invention aims to solve is the lack of research on ALK4 inhibitors in the prior art. Therefore, this invention provides a diaryl unsaturated ketone compound, its preparation method, an ALK4 inhibitor, and its applications.

[0005] To achieve the above objectives, the present invention provides a diaryl unsaturated ketone compound, the chemical structural formula of which is shown in Formula A:

[0006] Wherein: R1 and R2 are selected from hydrogen, deuterium, halogen, hydroxyl, amino, C1-C3 alkyl, C1-C3 alkoxy, -NH (C1-C3 alkyl), -N (C1-C3 alkyl)(C1-C3 alkyl), -C (=O)(C1-C3 alkyl), hydroxyl-substituted C1-C3 alkyl, 3-8 membered heterocyclic group, C1-C3 alkyl-substituted 3-8 membered heterocyclic group or halogen-substituted C1-C3 alkyl; The aromatic rings connected to R1 and R2 include one or more of the following: phenyl, naphthyl, pyridyl, pyrimidinyl, quinolinyl, furanyl, pyrroleyl, hexahydropyridyl, tetrahydropyranyl, tetrahydronaphthyl, furanyl, thiopheneyl, pyrroleyl, pyrazolyl, thiazolyl, pyrazinyl, pyridazinyl, 1,3,5-triazinyl, benzofuranyl, benzothiopheneyl, indolyl, inzolyl, benzimidazolyl, benzopyrazolyl, quinolinyl, isoquinolinyl, quinoxalinyl, benzopyrimidinyl, pyridopyrimidinyl, pyridopyridyl, purine, pteridinyl, imidazothiazolyl, imidazopyridazinyl, or pyrazopyrimidinyl.

[0007] Preferably, the chemical structural formula of the compound is any one of the following formulas: .

[0008] Under the same technical concept, the present invention also provides a method for preparing a diaryl unsaturated ketone compound, the synthetic route of which includes the following steps: (1) Add an alkaline salt, wherein the alkaline salt includes at least one of potassium carbonate, potassium hydroxide or sodium hydride, and then add a solvent, wherein the solvent includes at least one of methanol, ethanol, dimethylformamide or acetonitrile. Monitor the reaction by TLC until it is complete, add an extractant to extract, separate the organic phase, adjust the pH to 5-6, concentrate and dry to obtain a solid precursor. (2) The solid precursor was dry-loaded, purified by silica gel column chromatography, and separated by adding a mixture of methanol and dichloromethane; after the liquid was evaporated to dryness to obtain the solid, dichloromethane was added to dissolve the solid, and then petroleum ether was added to precipitate the solid. The solid was filtered and washed with a mixture of dichloromethane and petroleum ether to obtain a diaryl unsaturated or saturated ketone compound. Raw material I and raw material II contain R1 and R2, which are selected from hydrogen, deuterium, halogen, hydroxyl, amino, C1-C3 alkyl, C1-C3 alkoxy, -NH (C1-C3 alkyl), -N (C1-C3 alkyl)(C1-C3 alkyl), -C(=O)(C1-C3 alkyl), hydroxyl-substituted C1-C3 alkyl, 3-8 membered heterocyclic group, C1-C3 alkyl-substituted 3-8 membered heterocyclic group or halogen-substituted C1-C3 alkyl; R1 and R2 are linked to an aromatic ring, which includes one or more of the following: phenyl, naphthyl, pyridyl, pyrimidinyl, quinolinyl, furanyl, pyrroleyl, hexahydropyridyl, tetrahydropyranyl, tetrahydronaphthyl, furanyl, thiopheneyl, pyrroleyl, pyrazolyl, thiazolyl, pyrazinyl, pyridazinyl, 1,3,5-triazinyl, benzofuranyl, benzothiopheneyl, indolyl, inzolyl, benzimidazolyl, benzopyrazolyl, quinolinyl, isoquinolinyl, quinoxalinyl, benzopyrimidinyl, pyridopyrimidinyl, pyridopyridyl, purine, pteridinyl, imidazothiazolyl, imidazopyridazinyl, or pyrazopyrimidinyl.

[0009] The specific chemical reaction diagram is as follows:

[0010] Preferably, the preparation method further includes step (3), using the diaryl unsaturated or saturated ketone compound synthesized in step (2) as an intermediate, and continuing to add boron tribromide, with the addition amount in a molar ratio of 1-3:1 to the intermediate; adding a solvent, the solvent including at least one of 1,4-dioxane, dichloromethane, DMF or ethanol; reacting at room temperature; after the reaction is complete, concentrating, extracting and drying, and purifying by silica gel column chromatography to obtain the diaryl unsaturated or saturated ketone compound.

[0011] Preferably, the molar ratio of raw material I to raw material II is 0.1:1 to 1:10; The extractant includes at least one of saturated ammonium chloride, table salt, or dichloromethane; The molar ratio of potassium carbonate, potassium hydroxide, or sodium hydride to raw material I is 5-20 equivalents to 1 equivalent. The amount of solvent added is 0.1 to 1 molar concentration of the raw material.

[0012] Under the same technical concept, the present invention also provides an ALK4 inhibitor of a diaryl unsaturated ketone compound, wherein the diaryl unsaturated ketone compound is used to prepare the ALK4 inhibitor.

[0013] Preferably, the ALK4 inhibitor further includes the diaryl unsaturated ketone compound and its pharmacologically acceptable excipients or carriers.

[0014] Preferably, the ALK4 inhibitor is used to prevent or treat ALK4 receptor-mediated diseases including any one of melanoma, lung cancer, or breast cancer.

[0015] Under the same technical concept, the present invention also provides an ALK4 inhibitor of a diaryl unsaturated ketone compound, wherein the ALK4 inhibitor is used in diseases in which ALK4 is a key pathogenic signaling pathway.

[0016] Preferably, the ALK4 inhibitor can inhibit ALK4 enzyme activity and cell activity, tissue activity and model animal activity based on the enzyme function in a concentration range of 1 mg / kg to 100 mg / kg.

[0017] The above-described solution of the present invention has the following beneficial effects: (1) The diaryl unsaturated ketone compound with the chemical structure shown in Formula A provided by the present invention is a new type of skeleton among ALK4 inhibitors. This new structured compound can fill the gap in the systematic development of ALK4 inhibitors. (2) The diaryl unsaturated ketone compounds in this invention have excellent ALK4 inhibitory effects. The test results show that the specially modified compounds can specifically target ALK4 and have significant advantages in the treatment of the disease. (3) The synthesis method of the present invention is simple and convenient, and easy to prepare industrially. Attached Figure Description

[0018] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments or the prior art will be briefly introduced below. Obviously, the following drawings are some embodiments of the present invention. For those skilled in the art, other related drawings can be obtained from these drawings without creative effort.

[0019] Figure 1 This is a schematic diagram of the structure of the diaryl unsaturated ketone compound provided by the present invention; Figure 2 This is an activity inhibition diagram of the TZY29 diaryl unsaturated ketone compound provided in the embodiments of the present invention for ALK4 inhibition; Figure 3 , 4 The present invention provides nuclear magnetic resonance spectra of compounds represented by Example 3. Detailed Implementation

[0020] To make the technical problems, solutions, and advantages of this invention clearer, a detailed description will be provided below with reference to the accompanying drawings and specific embodiments. 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.

[0021] In the description of this invention, it should be noted that the terms "center," "upper," "lower," "left," "right," "vertical," "horizontal," "inner," and "outer," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are used only for the convenience of describing the invention and for simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on the invention. Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0022] In the description of this invention, it should be noted that, unless otherwise explicitly specified and limited, the terms "installation," "connection," and "linking" should be interpreted broadly. For example, they can refer to a locking connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; and they can refer to the internal connection of two components. Those skilled in the art can understand the specific meaning of the above terms in this invention based on the specific circumstances.

[0023] Furthermore, the technical features involved in the different embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0024] Figure 1 Here is a schematic diagram of the structure of the diaryl unsaturated ketone compound provided by the present invention; the structural formula of the diaryl unsaturated ketone compound provided in the embodiments of the present invention is as follows: .

[0025] The method for preparing diaryl saturated or unsaturated ketone compounds provided in this embodiment specifically includes:

[0026] Condition A: (1) Take I (0.25 mmol), II (0.25 mmol), and potassium carbonate (250 mg, 1.80 mmol) into a 25 ml round-bottom flask, add methanol (1 ml), and stir at room temperature. The reaction system gradually precipitates, and the reaction is complete as monitored by TLC. The reactants have completely reacted. After removing most of the solvent with a rotary evaporator, add hydrochloric acid to adjust the pH to 5-6, extract three times with dichloromethane, collect the organic layer, add anhydrous sodium sulfate to dry, and then evaporate to dryness to obtain the solid.

[0027] (2) Dry loading, silica gel column chromatography purification, eluent is dichloromethane:methanol=100:1. After the liquid is evaporated to dryness to obtain solid, dichloromethane (2 ml) is added to dissolve the solid, then petroleum ether (6 ml) is added to precipitate the solid. After filtration, the solid is washed 3 times with a dichloromethane:petroleum ether =1:3 mixed solution, and the filter residue is the product.

[0028] Example 1: The chemical formula of the diaryl unsaturated ketone compound in Example 1 is:

[0029] ( E )-5-methoxy-2-(3-methoxy-4-methylbenzylidene)-2,3-dihydro-1H-inden-1-one Using the aforementioned method for preparing diaryl unsaturated ketone compounds, the added raw material I is 4-methoxyepistone, and the added raw material II is 3-methoxy-4-methylbenzaldehyde.

[0030] The yield was 60.8 mg, with a yield of 82%. 1H NMR (500 MHz, Chloroform-d) δ 7.85 (d, J =8.4 Hz, 1H), 7.59 (t, J = 2.1 Hz, 1H), 7.23 – 7.19 (m, 2H), 7.07 (s, 1H),7.00 (d, J = 2.2 Hz, 1H), 6.96 (dd, J = 8.5, 2.3 Hz, 1H), 3.99 (s, 2H), 3.91(s, 3H), 3.90 (s, 3H), 2.27 (s, 3H). 13C NMR (101 MHz, Chloroform-d) δ192.55, 165.31, 154.27, 152.31, 151.77, 147.91, 147.80, 134.91, 134.89,132.27, 132.23, 131.79, 131.34, 126.25, 123.35, 123.28, 116.63, 116.44,115.76, 115.74, 115.35, 109.74, 56.33, 55.71, 32.31. HRMS (ESI), m / zcalculated for C 19 H 18 O3(M+H) + : 295.1334, found: 295.1343 Example 2: The chemical formula of the diaryl unsaturated ketone compound in Example 2 is:

[0031] ( E )-2-(4-aminobenzylidene)-5-methoxy-2,3-dihydro-1H-inden-1-one Using the aforementioned method for preparing diaryl unsaturated ketone compounds, the added raw material I is 4-methoxyepistone, and the added raw material II is 4-aminobenzaldehyde.

[0032] The yield was 56.3 mg, with a yield of 85%. 1 H NMR (400 MHz, DMSO- d6) δ 7.67 (d, J = 8.4 Hz,1H), 7.49 – 7.42 (m, 2H), 7.31 (t, J = 2.1 Hz, 1H), 7.16 (d, J = 2.2 Hz, 1H), 7.00 (dd, J = 8.4, 2.3 Hz, 1H), 6.68 – 6.62 (m, 2H), 5.87 (s, 2H), 3.95 (s,0H), 3.88 (s, 3H). 13 C NMR (101 MHz, DMSO) δ 191.97, 164.78, 152.79, 151.41,133.57, 133.23, 131.75, 129.76, 125.40, 122.77, 115.43, 114.29, 110.62,56.16, 32.72. HRMS (ESI), m / z calculated for C 17 H 15 NO2(M+H) + : 266.1181, found:266.1185 Example 3: The chemical formula of the diaryl unsaturated ketone compound in Example 3 is:

[0033] ( E )-2-(3-bromo-4-methoxybenzylidene)-5-methoxy-2,3-dihydro-1H-inden-1-one Using the aforementioned method for preparing diaryl unsaturated ketone compounds, the added raw material I is 4-methoxyepistone, and the added raw material II is 3-bromo-4-methoxybenzaldehyde. Figure 3 , 4 The present invention provides nuclear magnetic resonance spectra of compounds represented by Example 3.

[0034] The yield was 79.6 mg, with a yield of 89%. 1 H NMR (500 MHz, Chloroform- d ) δ 7.87 (d, J = 2.1Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.56 (dd, J= 8.6, 2.2 Hz, 1H), 7.49 (t, J = 2.1Hz, 1H), 7.01 (d, J = 2.2 Hz, 1H), 6.99 – 6.96 (m, 1H), 6.96 – 6.93 (m, 1H), 3.96 (s, 2H), 3.95 (s, 3H), 3.92 (s, 3H). 13 C NMR (126 MHz, Chloroform- d ) δ192.58, 165.25, 156.67, 152.31, 134.74, 134.19, 131.77, 131.44, 130.93,129.68, 126.19, 115.34, 112.28, 111.94, 109.72, 56.38, 55.72, 32.36. HRMS(ESI), m / z calculated for C 18 H 15 BrO3(M+H) + : 359.0283, found: 359.0288 Example 4: The chemical formula of the diaryl unsaturated ketone compound in Example 4 is:

[0035] ( E )-2-(3,4-dimethoxybenzylidene)-5-methoxy-2,3-dihydro-1H-inden-1-one Using the aforementioned method for preparing diaryl unsaturated ketone compounds, raw material I is 4-methoxyepistone, and raw material II is 3,4-dimethoxybenzaldehyde.

[0036] 69.7 mg of product was obtained, with a yield of 90%. 1H NMR (500 MHz, Chloroform- d ) δ 7.84 (d, J = 8.5Hz, 1H), 7.55 (t, J = 2.1 Hz, 1H), 7.28 (dd, J = 8.4, 2.0 Hz, 1H), 7.16 (d, J = 2.0Hz, 1H), 7.00 (d, J= 2.4 Hz, 1H), 6.95 (d, J = 4.8 Hz, 1H), 6.94 (d, J = 4.6 Hz,1H), 3.97 (s, 2H), 3.96 (s, 3H), 3.94 (s, 3H), 3.91 (s, 3H). 13 C NMR (126 MHz, Chloroform- d ) δ 192.79, 165.10, 152.26, 150.37, 149.06, 133.16, 132.83,131.62, 128.60, 126.10, 124.36, 115.18, 113.37, 111.29, 109.74, 55.99, 55.97,55.69, 32.45.HRMS (ESI), m / z calculated for C 19 H 18 O4(M+H) + : 311.1283, found: 311.1285 Example 5: The chemical formula of the diaryl unsaturated ketone compound in Example 5 is:

[0037] ( E )-2-(4-fluoro-3-methoxybenzylidene)-5-methoxy-2,3-dihydro-1H-inden-1-one Using the aforementioned method for preparing diaryl unsaturated ketone compounds, raw material I is 4-methoxyepistone, and raw material II is 3-methoxy-4-fluorobenzaldehyde.

[0038] The yield was 66.3 mg, with a yield of 89%. 1 H NMR (400 MHz, Chloroform- d ) δ 7.84 (d, J = 8.4Hz, 1H), 7.52 (t, J = 2.1 Hz, 1H), 7.26 – 7.17 (m, 2H), 7.19 – 7.09 (m, 1H), 6.99 (d, J = 2.2 Hz, 1H), 6.95 (dd, J= 8.5, 2.3 Hz, 1H), 3.95 (s, 3H), 3.94(s,2H), 3.91 (s, 3H). 13 C NMR (101 MHz, Chloroform- d ) δ 192.55, 165.31, 154.27,152.31, 151.77, 147.91, 147.80, 134.91, 134.89, 132.27, 132.23, 131.80,131.78, 131.34, 126.25, 123.35, 123.28, 116.63, 116.44, 115.76, 115.74,115.35, 109.74, 56.33, 55.71, 32.31. 19 F NMR (376 MHz, Chloroform- d ) δ -131.99.HRMS (ESI), m / z calculated for C 18 H 15 FO3(M+H) + : 299.1083, found: 299.1084 Example 6: The chemical formula of the diaryl unsaturated ketone compound in Example 6 is:

[0039] ( E )-2-(3,5-dihydroxybenzylidene)-5-methoxy-2,3-dihydro-1H-inden-1-one Using the aforementioned method for preparing diaryl unsaturated ketone compounds, the added raw material I is 4-methoxyepistone, and the added raw material II is 3,5-dihydroxybenzaldehyde.

[0040] The yield was 26.8 mg, with a yield of 38%. 1 H NMR (400 MHz, DMSO- d 6) δ 9.53 (s, 2H), 7.71 (d, J = 8.5 Hz, 1H), 7.25 – 7.20 (m, 1H), 7.17 (d, J = 2.2 Hz, 1H), 7.02 (dd, J =8.5, 2.3 Hz, 1H), 6.60 (d, J= 2.1 Hz, 2H), 6.32 (t, J = 2.1 Hz, 1H), 3.98 (s, 2H), 3.88 (s, 3H). 13 C NMR (101 MHz, DMSO- d 6) δ 192.12, 165.37, 159.13, 153.32,136.98, 135.59, 132.57, 131.02, 125.89, 115.82, 110.71, 109.16, 104.66,56.25, 32.54. HRMS (ESI), m / z calculated for C 17 H 14 O4(M+H) + : 283.0970, found:283.0973 Example 7: The chemical formula of the diaryl unsaturated ketone compound in Example 7 is:

[0041] ( E )-2-(3-bromo-5-ethoxy-4-hydroxybenzylidene)-5-methoxy-2,3-dihydro-1H-inden-1-one Using the aforementioned method for preparing diaryl unsaturated ketone compounds, the added raw material I is 4-methoxyepistone, and the added raw material II is 3-bromo-4-hydroxy-5-ethoxybenzaldehyde.

[0042] The yield was 70.8 mg, with a yield of 73%. 1 H NMR (400 MHz, DMSO- d 6) δ 9.91 (s, 1H), 7.70 (dd, J = 8.7, 3.7 Hz, 1H), 7.51 (d, J = 1.9 Hz, 1H), 7.35 (dt, J = 8.0, 2.8 Hz, 2H), 7.24 (d, J = 2.3 Hz, 1H), 7.02 (dd, J = 8.5, 2.3 Hz, 1H), 4.20 (q, J = 7.0 Hz,2H), 4.05 (s, 2H), 3.89 (d, J= 2.9 Hz, 4H), 1.40 (q, J = 5.3, 3.7 Hz, 4H). 13 C NMR (101 MHz, DMSO-) d 6) δ 191.94, 165.29, 153.35, 147.93, 146.07, 134.31, 131.32,131.09, 127.95, 127.40, 125.75, 115.96, 114.87, 110.58, 110.26, 65.14, 56.27,32.23, 14.98. HRMS (ESI), m / z calculated for C 19 H 17 BrO4(M+H) + : 389.0388, found: 389.0390 Example 8: The chemical formula of the diaryl unsaturated ketone compound in Example 8 is:

[0043] ( E )-2-(2-hydroxy-5-nitrobenzylidene)-5-methoxy-2,3-dihydro-1H-inden-1-one Using the aforementioned method for preparing diaryl unsaturated ketone compounds, raw material I is 4-methoxyepistone, and raw material II is 2-hydroxy-5-nitrobenzaldehyde.

[0044] The yield was 17.1 mg, with a yield of 22%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.51 (d, J = 2.8 Hz, 1H), 8.18 (dd, J = 9.0, 2.8 Hz, 1H), 7.77 – 7.69 (m, 2H), 7.29 (d, J = 2.2 Hz, 1H), 7.12 (d, J = 9.1 Hz, 1H), 7.03 (dd, J = 8.5, 2.3 Hz, 1H), 4.10 (d, J = 2.2 Hz, 2H), 4.10 (s, 0H), 3.89 (s, 3H). 13C NMR (101 MHz, DMSO) δ 191.86, 165.59,163.77, 153.46, 140.12, 137.27, 130.84, 127.03, 125.97, 125.51, 124.10,122.89, 116.72, 116.18, 110.72, 56.34, 32.07. HRMS (ESI), m / z calculated for C 17 H 13 NO5(M+H) + : 312.0872, found:312.0875 Example 9: The chemical formula of the diaryl unsaturated ketone compound in Example 9 is:

[0045] 5-methoxy-2-(pyridin-3-ylmethylene)-2,3-dihydro-1H-inden-1-one Using the aforementioned method for preparing diaryl unsaturated ketone compounds, the added raw material I is 4-methoxyepistone, and the added raw material II is pyridine-3-carboxaldehyde.

[0046] The yield was 52.1 mg, with a yield of 83%. 1 H NMR (500 MHz, Chloroform- d ) δ 8.69 (d, J = 2.3Hz, 1H), 8.55 (d, J = 2.4 Hz, 1H), 8.41 (dd, J = 4.8, 1.6 Hz, 1H), 8.36 (dd, J =4.8, 1.6 Hz, 1H), 7.73 (d, J = 8.6 Hz, 1H), 7.56 (ddt, J = 12.0, 10.0, 1.9 Hz,2H), 7.52 (d, J = 8.6 Hz, 1H), 7.10 (dddd, J = 16.1, 7.9, 4.8, 0.9 Hz, 2H), 6.95(dd, J = 8.6, 2.3 Hz, 1H), 6.68 (dd, J= 8.6, 2.3 Hz, 1H), 6.44 (d, J = 2.2 Hz, 1H), 6.40 (d, J = 2.2 Hz, 1H), 4.52 (dd, J = 10.7, 8.5 Hz, 1H), 4.04 (d, J = 10.8Hz, 1H), 3.93 (dd, J = 10.9, 8.5 Hz, 1H), 3.80 (d, J = 10.7 Hz, 1H), 3.71 (s,3H), 3.69 (s, 3H), 2.97 (d, J = 17.4 Hz, 1H), 2.90 (d, J = 17.4 Hz, 1H). 13 C NMR (126 MHz, Chloroform- d ) δ 204.08, 202.92, 165.77, 165.75, 157.62, 154.94,149.32, 149.26, 149.09, 148.55, 136.48, 136.05, 132.34, 132.07, 130.21,128.67, 126.65, 125.55, 123.32, 123.10, 115.91, 115.64, 109.12, 109.01,69.63, 56.67, 55.66, 55.53, 52.70, 51.86, 45.26, 29.39. HRMS (ESI), m / zcalculated for C 16 H 13 NO2(M+H) + : 251.0946, found: 252.1034 Example 10: The chemical formula of the diaryl unsaturated ketone compound in Example 10 is:

[0047] ( E )-5-methoxy-2-((4-methoxynaphthalen-1-yl)methylene)-2,3-dihydro-1H-inden-1-one Using the aforementioned method for preparing diaryl unsaturated ketone compounds, the added raw material I is 4-methoxyepistone, and the added raw material II is 4-methoxynaphthaldehyde.

[0048] The yield was 75.9 mg, with a yield of 92%. 1 H NMR (500 MHz, Chloroform- d ) δ 8.36 – 8.34(m, 1H), 8.34 – 8.31 (m, 1H), 8.24 (dt, J = 8.6, 0.9 Hz, 1H), 7.88 (dd, J = 8.1, 0.9 Hz, 1H), 7.75 (d, J = 8.1 Hz, 1H), 7.60 (ddd, J = 8.4, 6.8, 1.4 Hz, 1H), 7.53(ddd, J = 8.2, 6.8, 1.2 Hz, 1H), 6.98 – 6.93 (m, 2H), 6.89 (d, J = 8.1 Hz, 1H), 4.06 (s, 3H), 3.93 (d, J = 2.1 Hz, 2H), 3.89 (s, 3H). 13 C NMR (126 MHz, CDCl3) δ192.56, 165.05, 156.75, 152.77, 135.51, 133.49, 131.85, 129.49, 127.94,127.35, 126.19, 125.70, 125.63, 124.69, 123.79, 122.56, 115.08, 109.73,103.45, 55.67, 32.45.HRMS (ESI), m / z calculated for C 22 H 18 O3(M+H) + 331.1334, found: 331.1343 Example 11: The chemical formula of the diaryl unsaturated ketone compound in Example 11 is:

[0049] ( E)-2-(benzo[d][1,3]dioxol-4-ylmethylene)-5-methoxy-2,3-dihydro-1H-inden-1-one Using the aforementioned method for preparing diaryl unsaturated ketone compounds, raw material I is 4-methoxyepistone, and raw material II is 2,3-methylenedioxybenzaldehyde.

[0050] The yield was 56.6 mg, with a yield of 77%. 1 H NMR (500 MHz, Chloroform- d ) δ 7.84 (d, J = 8.5Hz, 1H), 7.69 (t, J = 2.2 Hz, 1H), 7.13 (dd, J = 8.0, 1.2 Hz, 1H), 6.97 (d, J = 2.3Hz, 1H), 6.94 (dd, J = 8.5, 2.2 Hz, 1H), 6.89 (t, J = 7.8 Hz, 1H), 6.83 (dd, J =7.8, 1.2 Hz, 1H), 6.05 (s, 2H), 3.95 (d, J = 2.2 Hz, 2H), 3.90 (s, 3H). 13 C NMR(126 MHz, CDCl3) δ 192.41, 165.25, 152.53, 147.84, 147.40, 136.27, 131.50,126.23, 125.31, 121.76, 121.74, 118.17, 115.23, 109.69, 109.29, 101.22,55.69, 32.47.HRMS (ESI), m / z calculated for C 18 H 14 O4(M+H) + : 295.0970, found:295.0977 Example 12: The chemical formula of the diaryl unsaturated ketone compound in Example 12 is:

[0051] ( E)-5-methoxy-2-(naphthalen-2-ylmethylene)-2,3-dihydro-1H-inden-1-one Using the aforementioned method for preparing diaryl unsaturated ketone compounds, raw material I is 4-methoxyepistone, and raw material II is 2-naphthaldehyde.

[0052] The yield was 72.0 mg, with a yield of 96%. 1 H NMR (500 MHz, Chloroform- d ) δ 8.13 (d, J = 1.7Hz, 1H), 7.94 – 7.83 (m, 4H), 7.81 – 7.75 (m, 2H), 7.57 – 7.50 (m, 2H), 7.04(d, J = 2.2 Hz, 1H), 6.98 (dd, J = 8.5, 2.3 Hz, 1H), 4.13 (d, J = 2.1 Hz, 2H), 3.93 (s, 3H). 13 C NMR (101 MHz, Chloroform- d ) δ 192.77, 165.27, 152.56, 135.49,133.50, 133.39, 133.15, 132.87, 131.54, 131.32, 128.61, 128.52, 127.71,127.25, 126.92, 126.67, 126.27, 115.30, 109.77, 55.72, 32.67. HRMS (ESI), m / zcalculated for C 21 H 16 O2(M+H) + 301.1228, found: 301.1228 Example 13: The chemical formula of the diaryl unsaturated ketone compound in Example 13 is:

[0053] ( E )-7-methoxy-2-(3-methoxy-4-methylbenzylidene)-3,4-dihydronaphthalen-1(2H)-one Using the aforementioned method for preparing diaryl unsaturated ketone compounds, the added raw material I is 7-methoxy-3,4-dihydronaphthone, and the added raw material II is 3-methoxy-4-methylbenzaldehyde.

[0054] The yield was 24.6 mg, with a yield of 32%. 1 H NMR (400 MHz, Chloroform- d ) δ 7.85 (d, J = 1.9Hz, 1H), 7.62 (d, J = 2.8 Hz, 1H), 7.21 – 7.13 (m, 2H), 7.07 (dd, J = 8.3, 2.8Hz, 1H), 6.98 (dd, J = 7.6, 1.6 Hz, 1H), 6.89 (d, J = 1.6 Hz, 1H), 3.88 (s, 3H), 3.86 (s, 3H), 3.14 (ddd, J = 6.8, 5.6, 1.9 Hz, 2H), 2.89 (dd, J = 7.5, 5.4 Hz, 2H), 2.25 (s, 3H). 13 C NMR (101 MHz, Chloroform- d ) δ 187.88, 158.65, 157.61,137.18, 135.92, 134.78, 134.63, 134.35, 130.50, 129.43, 127.75, 121.92,121.47, 111.59, 110.27, 55.58, 55.33, 28.05, 27.54, 16.27. HRMS (ESI), m / zcalculated for C 20 H 20 O3(M+H) + : 309.1490, found: 309.1498 Example 14: The chemical formula of the diaryl unsaturated ketone compound in Example 14 is:

[0055] ( E)-6-methoxy-2-(3-methoxy-4-methylbenzylidene)-3,4-dihydronaphthalen-1(2H)-one Using the aforementioned method for preparing diaryl unsaturated ketone compounds, the added raw material I is 6-methoxy-3,4-dihydronaphthone, and the added raw material II is 3-methoxy-4-methylbenzaldehyde.

[0056] 23.1 mg of product was obtained, with a yield of 30%. 1 H NMR (400 MHz, Chloroform- d ) δ 8.11 (d, J = 8.7Hz, 1H), 7.82 (d, J = 2.0 Hz, 1H), 7.17 (d, J = 7.6 Hz, 1H), 6.97 (dd, J = 7.7, 1.5Hz, 1H), 6.91 – 6.84 (m, 2H), 6.70 (d, J = 2.5 Hz, 1H), 3.87 (s, 3H), 3.85 (s,3H), 3.14 (td, J = 6.9, 6.5, 1.7 Hz, 2H), 2.91 (t, J = 6.5 Hz, 2H), 2.25 (s, 3H). 13 C NMR (101 MHz, Chloroform- d ) δ 186.84, 163.53, 157.59, 145.71, 136.42,134.94, 134.78, 130.75, 130.47, 127.53, 127.12, 121.82, 113.29, 112.27,111.56, 55.47, 55.33, 29.30, 27.38, 16.25. HRMS (ESI), m / z calculated forC 20 H 20 O3(M+H) + 309.1490, found: 309.1492 Example 15: The chemical formula of the diaryl unsaturated ketone compound in Example 15 is:

[0057] (E )-6,7-dimethoxy-2-(3-methoxy-4-methylbenzylidene)-3,4-dihydronaphthalen-1(2H)-one Using the aforementioned method for preparing diaryl unsaturated ketone compounds, the added raw material I is 6,7-dimethoxy-3,4-dihydronaphthone, and the added raw material II is 3-methoxy-4-methylbenzaldehyde.

[0058] The yield was 24.6 mg, with a yield of 32%. 1 H NMR (400 MHz, DMSO- d 6) δ 7.63 (s, 1H), 7.43 (s, 1H), 7.21 (d, J = 7.6 Hz, 1H), 7.06 (d, J = 1.6 Hz, 1H), 7.01 (dd, J = 7.6, 1.5Hz, 1H), 6.94 (s, 1H), 3.86 (s, 3H), 3.82 (s, 3H), 3.81 (s, 3H), 3.12 – 3.05(m, 2H), 2.88 (t, J = 6.5 Hz, 2H), 2.18 (s, 3H). 13 C NMR (101 MHz, DMSO- d 6) δ185.85, 157.69, 153.88, 148.33, 138.77, 135.65, 135.35, 134.82, 130.80,127.00, 126.18, 122.12, 112.36, 111.14, 109.43, 56.29, 55.94, 55.75, 28.10,27.54, 16.48. HRMS (ESI), m / z calculated for C 21 H 22 O3(M+H) + : 339.15960, found:339.1596 Example 16: The chemical formula of the diaryl unsaturated ketone compound in Example 16 is:

[0059] ( E)-2-(3-bromo-4,5-dihydroxybenzylidene)-5-methoxy-2,3-dihydro-1H-inden-1-one 1 H NMR (500 MHz, DMSO- d 6) δ 10.14 (s, 1H), 9.81 (s, 1H), 7.70 (d, J =8.4 Hz, 1H), 7.37 (d, J = 2.1 Hz, 1H), 7.25 (d, J = 2.3 Hz, 1H), 7.19 (t, J = 2.0Hz, 2H), 7.02 (dd, J = 8.5, 2.3 Hz, 1H), 3.98 (d, J = 2.2 Hz, 2H), 3.89 (s, 3H). 13 C NMR (126 MHz, DMSO- d 6) δ 191.90, 165.26, 153.08, 146.76, 145.37, 133.97,131.32, 131.16, 127.83, 126.52, 125.77, 116.78, 115.72, 110.75, 110.58,56.25, 32.37. HRMS (ESI), m / z calculated for C 17 H 13 B r O4(M+H) + : 361.0075, found:361.0080 Example 16 uses the product from Example 7 as an intermediate to continue synthesizing the product. The synthesis route is as follows: After anhydrous and oxygen-free conditions, a 25 ml round-bottom flask was prepared and the product from Example 7 (20 mg, 0.05 mmol) was added, dissolved in dichloromethane (0.5 ml). The flask was placed in an ice bath, and 1 M boron tribromide (0.15 mmol) was added. The reaction was allowed to proceed at room temperature. The reaction was monitored overnight by TLC to ensure complete reaction of the starting material. After quenching with methanol, some solvent was removed by rotary evaporation. The sample was then loaded onto a dry plate and purified by silica gel column chromatography, using dichloromethane:methanol = 40:1 as the eluent. The product from Example 16, 12.1 mg, was obtained, with a yield of 67%.

[0060] The diaryl unsaturated ketone compound obtained in the examples and the control substance LY364947 (CAS: 396129-53-6) were selected for bioactivity testing: Operating steps: Prepare 2x ATP / substrate solution and 2x kinase solution, and add kinase reaction buffer. Transfer 100 nL of the diaryl unsaturated ketone compound dilution to a 384 test plate using an Echo 655; centrifuge, add 5 μL of 2x kinase solution to the 384 test plate, centrifuge at 1000 rpm for 1 minute, and then incubate at 25°C for 10 minutes.

[0061] Add 5 μL of substrate and ATP solution to a 384-well plate and centrifuge at 1000 rpm for 10 minutes.

[0062] Rotate at 1 minute and then incubate at 25°C for 120 minutes.

[0063] Transfer 5 μL of ADP-Glo ​​to a 384 assay plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25 °C for 40 minutes.

[0064] Transfer 10 μL of the test solution to a 384 plate, centrifuge at 1000 rpm for 1 minute, and then incubate at 25°C for 40 minutes.

[0065] The luminescent signal was read using a multifunctional enzyme labeler.

[0066] The reagents and brand codes used in the examples are shown in Table 1 below: Table 1. Reagents and Brand Codes Used in the Examples

[0067] The reference standards are shown in Table 2 below: Table 2. ALK4 enzyme, substrate, and ATP control concentrations.

[0068] The inhibition rates of the embodiments in this application and the comparative experiments are as follows: Figure 2 As shown, the results are presented in Table 3 below: Table 3. ALK4 inhibition rate of the examples and comparative experiments

[0069] The inhibitory effects of diaryl unsaturated or saturated ketone compounds obtained in the examples on melanoma cell proliferation were determined by CCK-8 assay. A375, SK28, and SK5 cells frozen in liquid nitrogen were quickly placed in a 37°C water bath and agitated continuously to thaw rapidly. The thawed cell suspension was transferred to a centrifuge tube containing cell culture medium and centrifuged at 1000 r / min for 5 min, then the supernatant was discarded. 1 mL of complete culture medium was then added to the tube to resuspend the cells.

[0070] A375, SK28, and SK5 cells were passaged in DMEM medium containing 10% fetal bovine serum, 1% streptomycin, and 1% penicillin at 35 °C. A375, SK28, and SK5 cells in the logarithmic growth phase were collected for later use. Cells were digested with trypsin, and culture medium was added and mixed thoroughly by pipetting. 10 μl of the cell suspension was transferred to a cell counting plate, allowed to stand for a few minutes, and then the cells were counted under a microscope. Cell counting method: Number of cells required for seeding / Number of cells required per milliliter of culture medium = Required volume of stock medium.

[0071] Melanoma cells were seeded at a density of 3000 cells per well in 96-well plates. To reduce error, a ring of PBS or culture medium was added to the outermost layer of the 96-well plate. After cell attachment, the supernatant was replaced with 200 μL of Dulbecco modified Eagle (DMEM) medium containing 1 μM diaryl unsaturated ketone and 10% fetal bovine serum. Three replicates were made for each drug concentration, and the cells were cultured for another 48 hours. The supernatant was discarded, and 110 μL of CCK-8 working solution (medium and CCK-8 reagent were prepared at a ratio of 10:1) was added to each well. The cells were incubated at 37°C in a 5% CO2 incubator for 1.5-2 hours. The absorbance at 450 nm was read using a microplate reader, and cell viability was calculated (formula: cell viability = absorbance of drug-treated well / absorbance of control well × 100%).

[0072] The survival rates of the diaryl unsaturated ketones provided in the examples as ALK4 inhibitors are shown in Table 4 below: Table 4. Inhibitory effect of diaryl unsaturated ketone compounds on melanoma cell proliferation and survival rate as determined by the CCK-8 assay.

[0073] The present invention and its embodiments have been described above. This description is not restrictive, and the actual structure is not limited thereto. In conclusion, if those skilled in the art, inspired by this description, design similar structures and embodiments without departing from the spirit of the invention, such designs should fall within the scope of protection of this invention.

Claims

1. A diaryl unsaturated ketone compound, characterized in that, Its chemical structural formula is shown in Formula A: Wherein: R1 and R2 are selected from hydrogen, deuterium, halogen, hydroxyl, amino, C1-C3 alkyl, C1-C3 alkoxy, -NH (C1-C3 alkyl), -N (C1-C3 alkyl)(C1-C3 alkyl), -C (=O)(C1-C3 alkyl), hydroxyl-substituted C1-C3 alkyl, 3-8 membered heterocyclic group, C1-C3 alkyl-substituted 3-8 membered heterocyclic group or halogen-substituted C1-C3 alkyl; The aromatic rings connected to R1 and R2 include one or more of the following: phenyl, naphthyl, pyridyl, pyrimidinyl, quinolinyl, furanyl, pyrroleyl, hexahydropyridyl, tetrahydropyranyl, tetrahydronaphthyl, furanyl, thiopheneyl, pyrroleyl, pyrazolyl, thiazolyl, pyrazinyl, pyridazinyl, 1,3,5-triazinyl, benzofuranyl, benzothiopheneyl, indolyl, inzolyl, benzimidazolyl, benzopyrazolyl, quinolinyl, isoquinolinyl, quinoxalinyl, benzopyrimidinyl, pyridopyrimidinyl, pyridopyridyl, purine, pteridinyl, imidazothiazolyl, imidazopyridazinyl, or pyrazopyrimidinyl.

2. The compound according to claim 1, characterized in that, The chemical structural formula of the compound is any one of the following: 。 3. An ALK4 inhibitor of a diaryl unsaturated ketone compound, characterized in that, The diaryl unsaturated ketone compound is used to prepare ALK4 inhibitors.

4. The ALK4 inhibitor of the diaryl unsaturated ketone compound as described in claim 3, characterized in that, The ALK4 inhibitor also includes the diaryl unsaturated ketone compound and its pharmacologically acceptable excipients or carriers.

5. The ALK4 inhibitor of the diaryl unsaturated ketone compound as described in claim 3, characterized in that, The ALK4 inhibitors are used to prevent or treat any of the following ALK4 receptor-mediated diseases: melanoma, lung cancer, or breast cancer.

6. A method for preparing a diaryl unsaturated ketone compound, characterized in that, Includes the following steps: (1) Add an alkaline salt, wherein the alkaline salt includes at least one of potassium carbonate, potassium hydroxide or sodium hydride, and then add a solvent, wherein the solvent includes at least one of methanol, ethanol, dimethylformamide or acetonitrile. Monitor the reaction by TLC until it is complete, add an extractant to extract, separate the organic phase, adjust the pH to 5-6, concentrate and dry to obtain a solid precursor. (2) The solid precursor was dry-loaded, purified by silica gel column chromatography, and separated by adding a mixture of methanol and dichloromethane; after the liquid was evaporated to dryness to obtain the solid, dichloromethane was added to dissolve the solid, and then petroleum ether was added to precipitate the solid. The solid was filtered and washed with a mixed solution of dichloromethane and petroleum ether to obtain a diaryl unsaturated or saturated ketone compound. Raw material I and raw material II contain R1 and R2, which are selected from hydrogen, deuterium, halogen, hydroxyl, amino, C1-C3 alkyl, C1-C3 alkoxy, -NH (C1-C3 alkyl), -N (C1-C3 alkyl)(C1-C3 alkyl), -C(=O)(C1-C3 alkyl), hydroxyl-substituted C1-C3 alkyl, 3-8 membered heterocyclic group, C1-C3 alkyl-substituted 3-8 membered heterocyclic group or halogen-substituted C1-C3 alkyl; R1 and R2 are linked to an aromatic ring, which includes one or more of the following: phenyl, naphthyl, pyridyl, pyrimidinyl, quinolinyl, furanyl, pyrroleyl, hexahydropyridyl, tetrahydropyranyl, tetrahydronaphthyl, furanyl, thiopheneyl, pyrroleyl, pyrazolyl, thiazolyl, pyrazinyl, pyridazinyl, 1,3,5-triazinyl, benzofuranyl, benzothiopheneyl, indolyl, inzolyl, benzimidazolyl, benzopyrazolyl, quinolinyl, isoquinolinyl, quinoxalinyl, benzopyrimidinyl, pyridopyrimidinyl, pyridopyridyl, purine, pteridinyl, imidazothiazolyl, imidazopyridazinyl, or pyrazopyrimidinyl.

7. The preparation method according to claim 6, characterized in that, The preparation method further includes step (3), using the diaryl unsaturated or saturated ketone compound synthesized in step (2) as an intermediate, and continuing to add boron tribromide, with the addition amount in a molar ratio of 1-3:1 to the intermediate; adding a solvent, the solvent including at least one of 1,4-dioxane, dichloromethane, DMF or ethanol; reacting at room temperature; after the reaction is complete, concentrating, extracting and drying, and purifying by silica gel column chromatography to obtain the diaryl unsaturated or saturated ketone compound.

8. The application of an ALK4 inhibitor of a diaryl unsaturated ketone compound as described in any one of claims 3-5, characterized in that, The ALK4 inhibitors are used in diseases in which ALK4 is a key pathogenic signaling pathway.

9. The application as described in claim 8, characterized in that, The ALK4 inhibitor can inhibit ALK4 enzyme activity and cell, tissue and animal model activities based on the enzyme function in the concentration range of 1 mg / kg to 100 mg / kg.

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