Indole fused 4H-benzo [e] [1, 3] oxazine compound as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of organic synthesis, and particularly relates to an indole fused 4H-benzo [e] [1, 3] oxazine compound as well as a preparation method and application thereof. The structural formula of the compound is # imgabs0 #, wherein R1 is hydrogen, alkyl, alkoxy or halogen; r2 is aryl, alkoxy or alkyl; and R3 is benzyl, methyl or phenethyl. Compared with the existing preparation method of indole fused 4H-benzo [e] [1, 3] oxazine, the method disclosed by the invention has the advantages of no heavy metal or noble metal participation, smooth reaction at room temperature, low reaction energy consumption, simple operation, wide substrate range and low preparation cost. In addition, the prepared product 3e has the effect of obviously inhibiting Hela tumor cell proliferation, and can be used for preparing drugs for inhibiting tumor cell proliferation.

Description

Technical Field

[0001] The present invention belongs to the technical field of organic synthesis, and specifically relates to an indole-fused 4H-benzo[e][1,3]oxazine compound and a preparation method and application thereof. Background Art

[0002] Indole-fused 4H-benzo[e][1,3]oxazines are a unique class of heterocyclic aromatic skeletons that are present in the hepatitis C virus nonstructural protein NS5A inhibitor elbasvir, which can be used to treat chronic HCV infection (including co-infection with HIV).

[0003]

[0004] The existing preparation methods of indole-fused 4H-benzo[e][1,3]oxazine compounds require the participation of heavy metals or precious metals. There are currently three preparation methods: (1) using 2-benzyl-2H-benzo[e][1,3]oxazine derivatives as raw materials, under the catalysis of palladium acetate, 5 equivalents of potassium phosphate as additives, and obtaining them through intramolecular CN coupling at 55°C (J.Am.Chem.Soc.2015,137,13728-13731); (2) using N-(alkoxycarbonyl)-2-alkynyl indole as raw materials, in the presence of ferric chloride and diarylalkyl dihydrogen phosphate as raw materials, and obtaining them through intramolecular CN coupling at 55°C (J.Am.Chem.Soc.2015,137,13728-13731); It is obtained by intramolecular coupling at 70°C under the promotion of selenide (Chem. Eur. J. 2023, 29, e202202847); (3) N-alkoxy-1H-indole-1-carboxamide and propargyl alcohol are used as raw materials, catalyzed by metal rhodium, and an equivalent amount of copper acetate is used as an additive to obtain it by intermolecular [4+2] cyclization reaction at 70°C (Org. Lett. 2024, 26, 8680-8685).

[0005] The existing preparation methods of indole-fused 4H-benzo[e][1,3]oxazine compounds require the use of heavy metals (ferric chloride) or precious metal catalysts (palladium and rhodium), which have high preparation costs. The precious metal catalysts are difficult to completely remove after the reaction is completed, which limits the application of the products in the pharmaceutical field. In addition, the reaction requires a high temperature environment and high energy consumption, which limits the application of such methods. Summary of the invention

[0006] The technical problem to be solved by the present invention is to provide an indole-fused 4H-benzo[e][1,3]oxazine compound in view of the deficiencies in the prior art.

[0007] The problem that needs to be solved by the present invention is to provide a method for preparing the above-mentioned indole-fused 4H-benzo[e][1,3]oxazine compounds.

[0008] In order to solve the above technical problems, the present invention discloses an indole-fused 4H-benzo[e][1,3]oxazine compound, the structural formula of which is:

[0009]

[0010] Among them, R 1 is hydrogen, alkyl, alkoxy or halogen; R 2 is aryl, alkoxy or alkyl; R 3 is benzyl, methyl, phenethyl;

[0011] Preferably, R 1 is a hydrogen atom, a methyl group, an isopropyl group, a methoxy group, a fluorine atom, a chlorine atom or a bromine atom; R 2 is phenyl, methoxy, methyl, naphthyl; R 3 It is benzyl, methyl, and phenethyl.

[0012] The present invention also provides a method for preparing the above-mentioned indole-fused 4H-benzo[e][1,3]oxazine compounds, characterized in that the specific steps are: adding an indole-substituted phenol derivative, a catalyst and potassium carbonate to an organic solvent, mixing them evenly, adding 2-(acetoxymethyl)buta-2,3-dienoic acid esters while stirring, reacting at room temperature, and performing column chromatography separation on a silica gel column to obtain the indole-fused 4H-benzo[e][1,3]oxazine compounds.

[0013] Wherein, the structural formula of the indole-substituted phenol derivative is:

[0014]

[0015] Among them, R 1 is hydrogen, alkyl, alkoxy or halogen; R 2 is aryl, alkoxy or alkyl;

[0016] Preferably, R 1 is a hydrogen atom, a methyl group, an isopropyl group, a methoxy group, a fluorine atom, a chlorine atom or a bromine atom; R 2 It is phenyl, methoxy, methyl, naphthyl.

[0017] Wherein, the structural formula of the 2-(acetoxymethyl)butane-2,3-dienoic acid ester is:

[0018]

[0019] Among them, R 3 It is benzyl, methyl, and phenethyl.

[0020] Wherein, the molar ratio of the indole-substituted phenol derivative to the 2-(acetoxymethyl)buta-2,3-dienoic acid ester is 1:(1-2);

[0021] Preferably, the molar ratio of the indole-substituted phenol derivative to the 2-(acetoxymethyl)buta-2,3-dienoic acid ester is 1:1.5.

[0022] The added molar amount of the catalyst is 10-15% of the total molar amount of the indole-substituted phenol derivative and the 2-(acetoxymethyl)buta-2,3-dienoic acid ester;

[0023] Preferably, the added molar amount of the catalyst is 12% of the total molar amount of the indole-substituted phenol derivative and the 2-(acetoxymethyl)buta-2,3-dienoic acid ester;

[0024] More preferably, the catalyst is triethylenediamine.

[0025] Wherein, the eluent of the column chromatography separation is petroleum ether:ethyl acetate in a volume ratio of 10:1.

[0026] Furthermore, the use of the above-mentioned indole-fused 4H-benzo[e][1,3]oxazine compounds in the preparation of drugs for inhibiting tumor cell proliferation is also within the scope of protection of the present invention;

[0027] Specifically, in some embodiments of the present invention, a series of corresponding products, indole-fused 4H-benzo[e][1,3]oxazine compounds, are prepared by the above-mentioned preparation method, and the effects of some compounds on the activity of Hela cells are detected by MTT cell proliferation inhibition experiment. The experiment proves that the indole-fused 4H-benzo[e][1,3]oxazine compounds prepared by the present invention have the function of inhibiting the proliferation of tumor cells.

[0028] Beneficial effects: Compared with the existing preparation method of indole-fused 4H-benzo[e][1,3]oxazine, the method of the present invention does not require the participation of heavy metals or precious metals, can react smoothly at room temperature, has low reaction energy consumption, simple operation, a wide substrate range, and low preparation cost.

[0029] In addition, the prepared products of the present invention have the effect of inhibiting the proliferation of Hela tumor cells, among which product 3e has the best inhibitory effect. When the concentration of 3e is 64 μM, the survival rate of Hela cells is the lowest, which can reach 60.84% ​​at the lowest. It can be used to prepare drugs for inhibiting the proliferation of tumor cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] The present invention will be further described in detail below in conjunction with the accompanying drawings and specific embodiments, and the above and / or other advantages of the present invention will become more clear.

[0031] Figure 1The Hela cell toxicity test of compound 3a in Example 1;

[0032] Figure 2 The Hela cell toxicity test of compound 3e in Example 5;

[0033] Figure 3 This is the Hela cell cytotoxicity test of compound 3q in Example 17. DETAILED DESCRIPTION

[0034] The experimental methods described in the following examples are conventional methods unless otherwise specified; the reagents and materials described are commercially available unless otherwise specified.

[0035] The synthesis of indole substituted phenol derivatives in the following examples is based on Xu XH, Taniguchi M, Azuma A, et al. Remote anionic Fries rearrangement of sulfonates: Regioselective synthesis of indole triflones [J]. Organic Letters, 2013, 15 (3): 686-689.

[0036] The synthesis of 2-(acetoxymethyl)buta-2,3-dienoate in the following examples is based on Zhang Q, Yang L, Tong X. 2-(Acetoxymethyl)buta-2,3-dienoate, a Versatile 1,4-Biselectrophile for Phosphine-Catalyzed (4+n) Annulations with 1,n-Bisnucleophiles (n=1,2) [J]. Journal of the American Chemical Society, 2010, 132(8): 2550-2551.

[0037] Embodiment 1:

[0038] In this example, benzyl 2-(6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)acrylate was prepared, and its structural formula is as follows:

[0039]

[0040] The specific operation steps are as follows: at room temperature (25°C), 20.9 mg (0.10 mmol) of 2-(1H-indol-2-yl)phenol, 3.4 mg (0.03 mmol) of triethylenediamine, 20.7 mg (0.15 mmol) of potassium carbonate and 1.0 mL of chloroform are added to a 15 mL glass reaction tube in sequence, stirred evenly, and then 35.1 mg (0.15 mmol) of 2-(acetoxymethyl)but-2,3-dienoic acid benzyl ester is added under stirring, and then reacted at room temperature for 12 hours, and the reaction is monitored by TLC. After the reaction is completed, column chromatography is performed on a silica gel column (the eluent ratio is petroleum ether: ethyl acetate = 10:1, volume ratio) to obtain 32.3 mg of benzyl 2-(6-methyl-6H-benzo[5,6][1,3]oxazin[3,4-a]indol-6-yl)acrylate, recorded as 3a, as a white solid, with a yield of 82%.

[0041] The product data are characterized as follows:

[0042] Melting point 141~143℃;

[0043] 1 H NMR (400 MHz, CDCl 3 )δ7.62(dt,J=7.6,2.5Hz,2H),7.53–7.47(m,1H),7.33–7.26(m,3H),7.26–7.18(m,2H),7.18–7.09(m,3H ),7.03(td,J=7.5,1.2Hz,1H),6.86(d,J=8.3Hz,2H),6.11(s,1H),5.12(s,2H),4.75(s,1H),2.54(s,3H).

[0044] 13 C NMR (101 MHz, CDCl 3 )δ164.4,149.0,140.6,135.7,135.5,132.6,129.5,129.2,128.8,128.6,128.3,128 .2,123.7,123.0,122.6,121.1,120.6,118.2,117.6,111.9,97.6,90.6,66.7,26.8.

[0045] HRMS(ESI):m / z C 26 H 22 NO 3 ([M+H] + ):Calculated value: 396.1595; Measured value: 396.1596.

[0046] Example 2

[0047] In this example, 2-(6,10-dimethyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate was prepared, and its structural formula is as follows:

[0048]

[0049] The raw material 2-(1H-indol-2-yl)phenol was replaced with 2-(5-methyl-1H-indol-2-yl)phenol, and the rest was the same as in Example 1 to obtain 30.3 mg of the product 2-(6,10-dimethyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate, recorded as 3b, as a white solid with a yield of 74%.

[0050] The product data are characterized as follows:

[0051] Melting point 124~126℃;

[0052] 1 H NMR (400 MHz, CDCl 3 )δ7.61(dd,J=7.7,1.6Hz,1H),7.41(s,1H),7.39(d,J=8.7Hz,1H),7.35–7.2 8(m,3H),7.25(dd,J=6.5,2.5Hz,2H),7.14(td,J=7.7,1.6Hz,1H),7.03(td,J =7.6,1.2Hz,1H),6.97(dd,J=8.6,1.8Hz,1H),6.85(dd,J=8.1,1.1Hz,1H),6 .80(s,1H),6.09(s,1H),5.14(s,2H),4.72(s,1H),2.53(s,3H),2.43(s,3H).

[0053] 13 C NMR (101 MHz, CDCl 3 )δ164.4,148.9,140.6,135.6,133.9,132.7,129.9,129.7,129.0,128.7,128.5,128.2 ,128.1,124.2,123.5,122.9,120.7,118.3,117.6,111.6,97.1,90.5,66.7,26.7,21.3.

[0054] HRMS(ESI):m / z C 27 H 24 NO 3 ([M+H] +):Calculated value: 410.1751; Measured value: 410.1754.

[0055] Example 3

[0056] In this example, 2-(10-isopropyl-6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate was prepared, and its structural formula is as follows:

[0057]

[0058] The raw material 2-(1H-indol-2-yl)phenol was replaced with 2-(5-isopropyl-1H-indol-2-yl)phenol, and the rest was the same as in Example 1 to obtain 40.9 mg of the product 2-(10-isopropyl-6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate, recorded as 3c, as a white solid with a yield of 93%.

[0059] The product data are characterized as follows:

[0060] Melting point 129~131℃;

[0061] 1 H NMR (400 MHz, CDCl 3 )δ7.65–7.55(m,1H),7.46(s,1H),7.42(d,J=8.7Hz,1H),7.33–7.26(m ,3H),7.28–7.19(m,2H),7.17–7.09(m,1H),7.07–6.96(m,2H),6.85(d ,J=8.2Hz,1H),6.82(s,1H),6.09(s,1H),5.13(s,2H),4.74(s,1H),2. 99(p,J=6.9Hz,1H),2.53(s,3H),1.33–1.31(m,3H),1.31–1.29(m,3H).

[0062] 13 C NMR (101 MHz, CDCl 3 )δ164.5,148.9,141.2,140.6,135.7,134.1,132.7,129.6,128.9,128.9,128.6,128.2,128. 1,123.5,122.9,121.9,118.4,118.0,117.6,111.7,97.4,90.5,66.7,34.0,26.7,24.6,24.5.

[0063] HRMS (ESI) m / z C29 H 28 NO 3 ([M+H] + ):Calculated value: 438.2064; Measured value: 438.2067.

[0064] Example 4

[0065] In this example, 2-(10-methoxy-6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate was prepared, and its structural formula is as follows:

[0066]

[0067] The raw material 2-(1H-indol-2-yl)phenol was replaced with 2-(5-methoxy-1H-indol-2-yl)phenol, and the rest was the same as in Example 1 to obtain 29.1 mg of the product 2-(10-methoxy-6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate, recorded as 3d, as a white solid with a yield of 68%.

[0068] The product data are characterized as follows:

[0069] Melting point 129~131℃;

[0070] 1 H NMR (400 MHz, CDCl 3 )δ7.61(dd,J=7.7,1.6Hz,1H),7.40(d,J=9.2Hz,1H),7.36–7.29(m,3H),7.27–7.22(m,2H),7.14(td,J=7.7,1.6Hz,1H),7.08(d,J=2.6Hz,1H) ,7.03(td,J=7.5,1.2Hz,1H),6.86(dd,J=8.1,1.2Hz,1H),6.83–6.78(m ,2H),6.10(s,1H),5.13(s,2H),4.73(s,1H),3.85(s,3H),2.51(s,3H).

[0071] 13 C NMR (101 MHz, CDCl 3)δ164.4,154.5,148.9,140.6,135.6,133.2,130.7,130.1,129.0,128.7,128.5,128.2 ,128.1,123.5,122.9,118.2,117.6,112.6,112.6,102.5,97.2,90.4,66.7,55.7,26.7.

[0072] HRMS (ESI) m / z C 27 H 24 NO 4 ([M+H] + ):Calculated value: 426.1700; Measured value: 426.1702.

[0073] Example 5

[0074] In this example, 2-(10-chloro-6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate was prepared, and its structural formula is as follows:

[0075]

[0076] The raw material 2-(1H-indol-2-yl)phenol was replaced with 2-(5-chloro-1H-indol-2-yl)phenol, and the rest was the same as in Example 1 to obtain 37.0 mg of the product 2-(10-chloro-6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate, recorded as 3e, as a white solid with a yield of 86%.

[0077] The product data are characterized as follows:

[0078] Melting point 152~155℃;

[0079] 1 H NMR (400 MHz, CDCl 3 )δ7.60(dd,J=7.7,1.6Hz,1H),7.56(d,J=2.1Hz,1H),7.38(d,J=9.0Hz,1H),7.29(p,J=3.9Hz,3H),7.24–7.17(m,2H),7.16(dd,J=7.8,1. 6Hz,1H),7.05(qd,J=8.3,7.6,1.7Hz,2H),6.86(dd,J=8.2,1.1Hz,1H),6.78(s,1H),6.15(s,1H),5.10(s,2H),4.84(s,1H),2.47(s,3H).

[0080] 13C NMR (101 MHz, CDCl 3 )δ164.2,149.0,140.4,135.5,133.9,133.7,130.6,129.6,128.8,128.5,128.3, 128.2,126.3,123.8,123.1,122.7,120.3,117.7,112.8,97.0,90.4,66.8,26.5.

[0081] HRMS (ESI) m / z C 26 H 21 ClNO 3 ([M+H] + ):Calculated value: 430.1204; Measured value: 430.1208.

[0082] Example 6

[0083] In this example, 2-(10-bromo-6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate was prepared, and its structural formula is as follows:

[0084]

[0085] The raw material 2-(1H-indol-2-yl)phenol was replaced with 2-(5-bromo-1H-indol-2-yl)phenol, and the rest was the same as in Example 1 to obtain 35.9 mg of the product 2-(10-bromo-6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate, recorded as 3f, as a white solid with a yield of 76%.

[0086] The product data are characterized as follows:

[0087] Melting point 123~125℃;

[0088] 1 H NMR (400 MHz, CDCl 3 )δ7.72(d,J=2.0Hz,1H),7.60(dd,J=7.7,1.6Hz,1H),7.34(d,J=8.9Hz,1H),7.32–7.27(m,3H),7.24–7.13(m,4H),7.0 4(td,J=7.5,1.2Hz,1H),6.87(dd,J=8.1,1.2Hz,1H),6.78(s,1H),6.16(s,1H),5.10(s,2H),4.85(s,1H),2.46(s,3H).

[0089] 13C NMR (101 MHz, CDCl 3 )δ164.2,149.0,140.4,135.5,134.0,133.7,131.2,129.6,128.8,128.5,128.3,128 .2,125.2,123.8,123.4,123.1,117.7,117.6,113.9,113.2,96.8,90.4,66.8,26.5.

[0090] HRMS (ESI) m / z C 26 H 21 BrNO 3 ([M+H] + ):Calculated value: 474.0699; Measured value: 474.0703.

[0091] Example 7

[0092] In this example, 2-(9-chloro-6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate was prepared, and its structural formula is as follows:

[0093]

[0094] The raw material 2-(1H-indol-2-yl)phenol was replaced with 2-(6-chloro-1H-indol-2-yl)phenol, and the rest was the same as in Example 1 to obtain 30.6 mg of the product 2-(9-chloro-6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate, recorded as 3 g, as a white solid, with a yield of 71%.

[0095] The product data are characterized as follows:

[0096] Melting point 149~151℃;

[0097] 1 H NMR (400 MHz, CDCl 3)δ7.60(dd,J=7.7,1.7Hz,1H),7.52(d,J=8.5Hz,1H),7.50–7.48(m,1H),7. 36–7.28(m,3H),7.28–7.22(m,2H),7.17(td,J=7.7,1.6Hz,1H),7.10(dd,J =8.4,1.8Hz,1H),7.05(td,J=7.5,1.2Hz,1H),6.87(dd,J=8.1,1.2Hz,1H), 6.83(d,J=0.9Hz,1H),6.16(s,1H),5.13(s,2H),4.81(s,1H),2.50(s,3H).

[0098] 13 C NMR (101 MHz, CDCl 3 )δ164.1,148.9,140.3,135.6,135.5,133.4,129.4,128.9,128.5,128.3,128.2,128 .2,128.0,123.6,123.1,121.7,121.3,117.8,117.7,111.8,97.5,90.4,66.8,26.6.

[0099] HRMS (ESI) m / z C 26 H 21 ClNO 3 ([M+H] + ):Calculated value: 430.1204; Measured value: 430.1208.

[0100] Example 8

[0101] In this example, 2-(9-bromo-6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate was prepared, and its structural formula is as follows:

[0102]

[0103] The raw material 2-(1H-indol-2-yl)phenol was replaced with 2-(6-bromo-1H-indol-2-yl)phenol, and the rest was the same as in Example 1 to obtain 31.8 mg of the product 2-(9-bromo-6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate, recorded as 3h, as a white solid, with a yield of 67%.

[0104] The product data are characterized as follows:

[0105] Melting point 168~170℃;

[0106] 1 H NMR (400 MHz, CDCl 3 )δ7.66–7.64(m,1H),7.60(dd,J=7.7,1.6Hz,1H),7.47(d,J=8.4Hz,1H),7.35–7.28(m,3H),7.27–7.22(m,3H),7.17(td,J=7.7,1.6Hz,1 H),7.05(td,J=7.5,1.2Hz,1H),6.86(dd,J=8.2,1.2Hz,1H),6.83(d,J=0.8Hz,1H),6.15(s,1H),5.13(s,2H),4.79(s,1H),2.50(s,3H).

[0107] 13 C NMR (101 MHz, CDCl 3 )δ164.1,148.9,140.2,136.0,135.5,133.3,129.5,128.9,128.6,128.3,128.3,128 .2,123.9,123.7,123.1,122.1,117.8,117.7,116.0,114.7,97.5,90.4,66.8,26.6.

[0108] HRMS (ESI) m / z C 26 H 21 BrNO 3 ([M+H] + ):Calculated value: 474.0699; Measured value: 474.0704.

[0109] Example 9

[0110] In this example, 2-(8-fluoro-6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate was prepared, and its structural formula is as follows:

[0111]

[0112] The raw material 2-(1H-indol-2-yl)phenol was replaced with 2-(7-fluoro-1H-indol-2-yl)phenol, and the rest was the same as in Example 1 to obtain 38.1 mg of the product 2-(8-fluoro-6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate, recorded as 3i, as a white solid with a yield of 92%.

[0113] The product data are characterized as follows:

[0114] Melting point 126~128℃;

[0115] 1 H NMR (400 MHz, CDCl 3 )δ7.57(dd,J=7.7,1.6Hz,1H),7.38(d,J=7.8Hz,1H),7.32–7.26(m,3H),7.24–7.20(m,2H),7.17(td,J=7.8,1 .6Hz,1H),7.07–6.99(m,2H),6.90–6.82(m,3H),6.25(s,1H),5.19(s,1H),5.11(s,2H),2.46(d,J=3.2Hz,3H).

[0116] 13 C NMR (101 MHz, CDCl 3 )δ164.4,148.8,148.5(d,J=245.0Hz),140.9(d,J=2.0Hz),135.7,133.7(d,J=4.8Hz),133.6,129.6,128.8(d,J=2.1Hz),128.5,128.2,128.2 ,123.9,122.9,121.2(d,J=7.3Hz),117.4,117.4,116.7(d,J=3.3Hz),109.1(d,J=22.2Hz),98.0(d,J=2.1Hz),91.1,66.7,27.6(d,J=15.8Hz).

[0117] HRMS (ESI) m / z C 26 H 21 FNO 3 ([M+H] + ):Calculated value: 414.1500; Measured value: 414.1502.

[0118] Example 10

[0119] In this example, 2-(8-fluoro-6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate was prepared, and its structural formula is as follows:

[0120]

[0121] The raw material 2-(1H-indol-2-yl)phenol was replaced with 2-(7-chloro-1H-indol-2-yl)phenol, and the rest was the same as in Example 1 to obtain 31.6 mg of the product 2-(8-chloro-6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate, recorded as 3j, as a white solid with a yield of 73%.

[0122] The product data are characterized as follows:

[0123] Melting point 190~192℃;

[0124] 1 H NMR (400 MHz, CDCl 3 )δ7.53(dd,J=17.2,7.7Hz,2H),7.35–7.27(m,5H),7.25–7.11(m,2H),7.04(dt,J=11.1,7.6Hz,2H) ,6.89(d,J=8.1Hz,1H),6.85(s,1H),6.38(s,1H),5.50(s,1H),5.20(d,J=4.5Hz,2H),2.57(s,3H).

[0125] 13 C NMR (101 MHz, CDCl 3 )δ164.6,148.2,141.5,135.8,133.7,133.3,132.1,130.0,129.7,128.6,128.2,128 .1,125.3,124.1,123.1,121.6,119.7,118.1,117.5,116.4,98.1,91.2,66.7,29.2.

[0126] HRMS (ESI) m / z C 26 H 21 ClNO 3 ([M+H] + ):Calculated value: 430.1204; Measured value: 430.1206.

[0127] Embodiment 11

[0128] In this example, 2-(6,9,11-trimethyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate was prepared, and its structural formula is as follows:

[0129]

[0130] The raw material 2-(1H-indol-2-yl)phenol was replaced with 2-(4,6-dimethyl-1H-indol-2-yl)phenol, and the rest was the same as in Example 1 to obtain 27.1 mg of the product 2-(6,9,11-trimethyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate, recorded as 3k, as a white solid with a yield of 64%.

[0131] The product data are characterized as follows:

[0132] Melting point 143~145℃;

[0133] 1 H NMR (400 MHz, CDCl 3 )δ7.63(dd,J=7.6,1.6Hz,1H),7.36–7.29(m,3H),7.28–7.21(m,2H),7.14(s,1H),7.11(dd,J=7.9,1.6Hz,1H),7 .03(td,J=7.5,1.2Hz,1H),6.88–6.76(m,3H),6.11(s,1H),5.15(s,2H),4.73(s,1H),2.54(s,6H),2.41(s,3H).

[0134] 13 C NMR (101 MHz, CDCl 3 )δ164.4,148.7,140.5,135.7,135.7,132.5,131.5,130.0,128.9,128.7,128.5,128.2,12 8.2,127.1,123.3,122.9,122.7,118.5,117.6,109.6,96.0,90.5,66.7,26.8,22.2,18.8.

[0135] HRMS (ESI) m / z C 28 H 26 NO 3 ([M+H] + ):Calculated value: 424.1907; Measured value: 424.1908.

[0136] Example 12

[0137] In this example, 2-(6-methyl-2-phenyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate was prepared, and its structural formula is as follows:

[0138]

[0139] The raw material 2-(1H-indol-2-yl)phenol was replaced with 3-(1H-indol-2-yl)-[1,1'-biphenyl]-4-ol, and the rest was the same as in Example 1 to obtain 37.7 mg of the product 2-(6-methyl-2-phenyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate, recorded as 31, as a white solid with a yield of 80%.

[0140] The product data are characterized as follows:

[0141] Melting point 151~153℃;

[0142] 1 H NMR (400 MHz, CDCl 3 )δ7.83(d,J=2.2Hz,1H),7.66–7.62(m,1H),7.61–7.56(m,2H),7.55–7.48(m,1H),7.47–7.41(m,2H),7.40–7.33(m,2H),7.32–7.26( m,3H),7.27–7.21(m,2H),7.20–7.10(m,2H),6.94(s,1H),6.92(d,J=8.5Hz,1H),6.15(s,1H),5.14(s,2H),4.81(s,1H),2.55(s,3H).

[0143] 13 C NMR (101 MHz, CDCl 3 )δ164.4,148.5,140.6,140.6,136.2,135.6,135.5,132.5,129.5,128.9,128.9,128.6,128.3,128 .2,128.0,127.2,127.0,122.7,122.2,121.2,120.7,118.3,117.9,111.9,97.8,90.7,66.8,26.8.

[0144] HRMS (ESI) m / z C 32 H 26 NO 3 ([M+H] + ):Calculated value: 472.1907; Measured value: 472.1912.

[0145] Example 13

[0146] In this example, 2-(3-methoxy-6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate was prepared, and its structural formula is as follows:

[0147]

[0148] The raw material 2-(1H-indol-2-yl)phenol was replaced with 2-(1H-indol-2-yl)-5-methoxyphenol, and the rest was the same as in Example 1 to obtain 25.9 mg of the product 2-(3-methoxy-6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate, recorded as 3m, as a white solid with a yield of 61%.

[0149] The product data are characterized as follows:

[0150] Melting point 128~130℃;

[0151] 1 H NMR (400 MHz, CDCl 3 )δ7.63–7.57(m,1H),7.54(d,J=8.6Hz,1H),7.48(dq,J=6.7,3.9Hz,1H),7.38–7.27(m,4H),7.27–7.20(m,2H),7.15–7.07(m,2H),6.75 (d,J=0.8Hz,1H),6.64(dd,J=8.6,2.5Hz,1H),6.46(d,J=2.5Hz,1H),6.14(s,1H),5.15(s,2H),4.75(s,1H),3.73(s,3H),2.53(s,3H).

[0152] 13 C NMR (101 MHz, CDCl 3 )δ164.3,160.7,150.2,140.3,135.7,135.2,132.9,129.7,128.9,128.5,128.2,128.0 ,124.6,122.0,120.7,120.5,111.7,111.0,110.2,102.3,95.9,90.6,66.6,55.5,26.7.

[0153] HRMS (ESI) m / z C 27 H 24 NO 4 ([M+H] + ):Calculated value: 426.1700; Measured value: 426.1702.

[0154] Embodiment 14

[0155] In this example, 2-(4,6-dimethyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate was prepared, and its structural formula is as follows:

[0156]

[0157] The raw material 2-(1H-indol-2-yl)phenol was replaced with 2-(1H-indol-2-yl)-6-methylphenol, and the rest was the same as in Example 1 to obtain 31.5 mg of the product 2-(4,6-dimethyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate, recorded as 3n, as a white solid with a yield of 77%.

[0158] The product data are characterized as follows:

[0159] Melting point 183~186℃;

[0160] 1 H NMR (400 MHz, CDCl 3 )δ7.67–7.60(m,1H),7.55–7.45(m,2H),7.35–7.27(m,3H),7.27–7.20(m,2H),7.19–7.09(m,2H),7.03(ddd,J=7.4,1.7,0.9Hz ,1H),6.95(t,J=7.5Hz,1H),6.87(d,J=0.8Hz,1H),6.06(s,1H),5.13(q,J=12.4Hz,2H),4.61(s,1H),2.61(s,3H),2.16(s,3H).

[0161] 13 C NMR (101 MHz, CDCl 3 )δ164.3,147.2,140.4,135.6,135.4,133.1,130.5,129.5,128.5,128.3,128.1,12 7.2,122.5,122.4,121.2,121.0,120.5,117.7,111.9,97.5,90.5,66.6,27.0,15.3.

[0162] HRMS (ESI) m / z C 27 H 24 NO 3 ([M+H] + ):Calculated value: 410.1751; Measured value: 410.1752.

[0163] Embodiment 15

[0164] In this example, 2-(1,3,6-trimethyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate was prepared, and its structural formula is as follows:

[0165]

[0166] The raw material 2-(1H-indol-2-yl)phenol was replaced with 2-(1H-indol-2-yl)-3,5-dimethylphenol, and the rest was the same as in Example 1 to obtain 32.6 mg of the product 2-(1,3,6-trimethyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)benzyl acrylate, recorded as 3o, as a white solid with a yield of 95%.

[0167] The product data are characterized as follows:

[0168] Melting point 139~141℃;

[0169] 1 H NMR (400 MHz, CDCl 3 )δ7.68–7.62(m,1H),7.56–7.50(m,1H),7.37–7.27(m,3H),7.24(dd,J=7.3,2.3Hz,2H),7.14(ttd,J=7.2,5.7,1.3Hz,2H ),6.88(s,1H),6.73(s,1H),6.58(s,1H),6.07(s,1H),5.13(s,2H),4.75(s,1H),2.56(s,3H),2.54(s,2H),2.23(s,3H).

[0170] 13 C NMR (101 MHz, CDCl 3 )δ164.5,149.8,140.8,138.6,135.7,135.1,134.4,132.0,129.4,128.7,128.6,128.3,12 8.3,126.9,122.5,121.0,120.3,116.0,115.0,111.8,101.8,89.7,66.7,27.1,22.5,21.4.

[0171] HRMS (ESI) m / z C 28 H 26 NO 3 ([M+H] + ):Calculated value: 424.1907; Measured value: 424.1910.

[0172] Example 16

[0173] In this example, 2-(8-methyl-8H-naphtho[1',2':5,6][1,3]oxazino[3,4-a]indol-8-yl)benzyl acrylate was prepared, and its structural formula is as follows:

[0174]

[0175] The raw material 2-(1H-indol-2-yl)phenol was replaced with 1-(1H-indol-2-yl)naphthalen-2-ol, and the rest was the same as in Example 1 to obtain 41.6 mg of the product 2-(8-methyl-8H-naphtho[1',2':5,6][1,3]oxazino[3,4-a]indol-8-yl)benzyl acrylate, recorded as 3p, as a white solid with a yield of 93%.

[0176] The product data are characterized as follows:

[0177] Melting point 184~186℃;

[0178] 1 H NMR (400 MHz, CDCl 3 )δ8.63(d,J=8.6Hz,1H),7.80(d,J=8.1Hz,1H),7.71(dt,J=8.2,1.4Hz,1H),7.64(d,J=8.8Hz,1H),7.62–7.55(m,2H),7.46–7.38(m,1H), 7.31–7.24(m,4H),7.24–7.13(m,4H),7.07(dd,J=8.8,1.5Hz,1H),6.07(s,1H),5.09(d,J=2.8Hz,2H),5.01(d,J=1.1Hz,1H),2.64(s,3H).

[0179] 13 C NMR (101 MHz, CDCl 3 )δ164.4,148.4,140.8,135.6,134.4,130.9,130.7,130.2,129.6,129.6,129.1,129.1,128.6, 128.3,128.2,127.5,124.6,124.3,122.8,121.2,120.5,118.4,111.9,102.0,89.9,66.7,27.0.

[0180] HRMS (ESI) m / z C 30 H 24 NO 3 ([M+H] +):Calculated value: 446.1751; Measured value: 446.1754.

[0181] Embodiment 17

[0182] In this example, methyl 2-(6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)acrylate was prepared, and its structural formula is as follows:

[0183]

[0184] The raw material 2-(acetoxymethyl)but-2,3-dienoic acid benzyl ester was replaced with 2-(acetoxymethyl)but-2,3-dienoic acid methyl ester, and the rest was the same as in Example 1 to obtain 23.3 mg of the product methyl 2-(6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)acrylate, recorded as 3q, as a white solid with a yield of 73%.

[0185] The product data are characterized as follows:

[0186] Melting point 128~130℃;

[0187] 1 H NMR (400 MHz, CDCl 3 )δ7.64(ddd,J=5.4,3.9,1.8Hz,2H),7.55–7.49(m,1H),7.26–7.10(m,3H),7.09–7.02(m,1 H),6.99(d,J=8.1Hz,1H),6.89(s,1H),6.06(s,1H),4.72(s,1H),3.67(s,3H),2.55(s,3H).

[0188] 13 C NMR (101 MHz, CDCl 3 )δ164.9,149.0,140.3,135.5,132.6,129.4,129.2,128.5,123.6,123.0,122.6,121.1,120.6,118.2,117.6,111.9,97.6,90.5,52.1,26.8.

[0189] HRMS (ESI) m / z C 20 H 18 NO 3 ([M+H] + ):Calculated value: 320.1281; Measured value: 320.1283.

[0190] Embodiment 18

[0191] In this example, 2-(6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)phenethyl acrylate was prepared, and its structural formula is as follows:

[0192]

[0193] The raw material 2-(acetoxymethyl)but-2,3-dienoic acid benzyl ester was replaced with 2-(acetoxymethyl)but-2,3-dienoic acid phenethyl ester, and the rest was the same as in Example 1 to obtain 32.8 mg of the product 2-(6-methyl-6H-benzo[5,6][1,3]oxazino[3,4-a]indol-6-yl)phenethyl acrylate, recorded as 3r, as a white solid with a yield of 80%.

[0194] The product data are characterized as follows:

[0195] Melting point 155~157℃;

[0196] 1 H NMR (400 MHz, CDCl 3 )δ7.63(ddd,J=6.1,4.7,1.9Hz,1H),7.52–7.45(m,1H),7.29–7.09(m,4H),7.05(td,J=7.5,1.2Hz,1H),6.93(dd,J =8.1,1.1Hz,0H),6.88(s,0H),6.04(s,1H),4.74(s,1H),4.28(t,J=7.0Hz,1H),2.83(t,J=7.0Hz,1H),2.49(s,1H).

[0197] 13 C NMR (101 MHz, CDCl 3 )δ164.5,149.0,140.5,137.7,135.5,132.6,129.5,129.2,128.9,128.7,128.5,126.6 ,123.7,123.0,122.6,121.1,120.6,118.2,117.6,111.9,97.6,90.5,65.5,34.9,26.7.

[0198] HRMS (ESI) m / z C 27 H 24 NO 3 ([M+H] + ):Calculated value: 410.1751; Measured value: 410.1754.

[0199] The proliferation inhibition activity of some of the compounds in the examples was tested on several tumor cell lines. The experimental method is as follows:

[0200] The tumor cells (Hela cells) used in the experiment were from the Cell Resource Center of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. The MTT cell proliferation inhibition assay was used to detect the effects of the compounds on the activity of Hela cells. Three types of cells growing in the logarithmic phase were taken, and Hela cells were digested with trypsin to obtain a cell suspension, which was counted and adjusted to a cell density of 2×10 4 200 μL of cells were seeded into each well of a 96-well plate and incubated at 37°C and 5% CO 2 After culturing in an incubator for 24 hours, different examples of compounds (3a, 3e, 3q) were added in sequence at concentrations of 1 μM, 2 μM, 4 μM, 8 μM, 16 μM, 32 μM, and 64 μM. DMSO was used as the solvent. Three replicates were added to each compound. The blank control group was only added with the corresponding volume of solvent (DMSO) without adding the compound. Then, the cells were placed at 37°C and 5% CO. 2 Incubate in an incubator for 48 hours. Then add 20 μL of 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT) at a final concentration of 5 mg / mL and incubate for 4 hours. Use a Bio-Rad microplate reader to detect the absorbance value of each well at a wavelength of 490 nm.

[0201] Cell number (% blank group) = 100 × (absorbance value of compound treatment group - absorbance value of blank group) / (absorbance value of control group - absorbance value of blank group)

[0202] Each experiment was repeated three times, with compound concentration as the horizontal axis and cell survival rate as the vertical axis, and the graph was drawn using GraphPadPrism software.

[0203] Figure 1 , Figure 2 , Figure 3 The results of the test on the proliferation inhibition activity of the indole-fused 4H-benzo[e][1,3]oxazine compounds of Example 1, Example 5 and Example 17 on Hela cells are shown in Table 1, Table 2 and Table 3 respectively:

[0204] Table 1

[0205] 3a(concentration / μM) Hela cell survival rate (%) 1 97.51 2 97.44 4 98.73 8 99.40 16 100.69 32 98.95 64 90.69 Blank control group 100

[0206] Table 2

[0207] 3e(concentration / μM) Hela cell survival rate (%) 1 85.96 2 87.28 4 66.88 8 67.97 16 63.96 32 64.75 64 60.84 Blank control group 100

[0208] Table 3

[0209]

[0210]

[0211] The present invention provides an indole-fused 4H-benzo[e][1,3]oxazine compound and a preparation method and application thereof. There are many methods and approaches to implement the technical solution. The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention. All components not specified in this embodiment can be implemented by existing technologies.

Claims

1. An indole-fused 4H-benzo[e][1,3]oxazine compound, characterized in that: The compound structural formula is: Among them, R 1 is hydrogen, alkyl, alkoxy or halogen; R 2 is aryl, alkoxy or alkyl; R 3 It is benzyl, methyl, and phenethyl.

2. The method for preparing the indole-fused 4H-benzo[e][1,3]oxazine compound according to claim 1, characterized in that: The specific steps are: adding an indole-substituted phenol derivative, a catalyst and potassium carbonate to an organic solvent, mixing them evenly, adding 2-(acetoxymethyl)but-2,3-dienoic acid esters while stirring, reacting at room temperature, and performing column chromatography separation on a silica gel column to obtain the indole-fused 4H-benzo[e][1,3]oxazine compound.

3. The preparation method according to claim 2, characterized in that: The structural formula of the indole substituted phenol derivative is: Among them, R 1 is hydrogen, alkyl, alkoxy or halogen; R 2 is aryl, alkoxy or alkyl.

4. The preparation method according to claim 2, characterized in that: The structural formula of the 2-(acetoxymethyl)buta-2,3-dienoic acid esters is: Among them, R 3 It is benzyl, methyl, and phenethyl.

5. The preparation method according to claim 2, characterized in that: The molar ratio of the indole-substituted phenol derivative to the 2-(acetoxymethyl)buta-2,3-dienoic acid ester is 1:(1-2).

6. The preparation method according to claim 2, characterized in that: The added molar amount of the catalyst is 10-15% of the total molar amount of the indole-substituted phenol derivative and the 2-(acetoxymethyl)buta-2,3-dienoic acid ester.

7. The preparation method according to claim 6, characterized in that: The catalyst is triethylenediamine.

8. The preparation method according to claim 2, characterized in that: The column chromatography separation uses a eluent of petroleum ether:ethyl acetate in a volume ratio of 10:

1.

9. Use of the indole-fused 4H-benzo[e][1,3]oxazine compound of claim 1 in the preparation of a drug for inhibiting tumor cell proliferation.

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