A method for preparing N-triazine amide compounds

Abstract

The present invention provides a method for preparing N-triazine amide compounds. This process is convenient to operate and the raw materials are easily available, having certain application value. The N-triazine amide compounds provided by the present invention exhibit certain anti-hepatocarcinoma and anti-breast cancer biological activities, laying a foundation for new drug screening and development.

Description

(1) Technical Field

[0001] The present invention relates to a method for preparing N-triazine amide compounds. (2) Background Art

[0002] Amide compounds are important organic synthesis units and also the basic skeletons of many drugs and natural products. The traditional amide bond is mainly prepared by the condensation reaction of acyl chloride or carboxylic acid with amine, but this method has disadvantages such as easy decomposition of raw materials used, difficult removal of by-products, difficult control of reaction conditions, cumbersome post-treatment, and poor yield, and is subject to certain limitations in use. Triazine compounds have various biological activities such as anti-cancer, antibacterial, anti-malarial, and herbicidal effects, which are one of the research hotspots of scientists in recent years. Therefore, developing a method for preparing N-triazine amide compounds has certain practical application value. (3) Summary of the Invention

[0003] The present invention provides a method for preparing N-triazine amide compounds.

[0004] The present invention adopts the following technical solutions:

[0005] The present invention provides a method for preparing an N-triazine amide compound represented by formula (I), and the method is as follows:

[0006] Mix a triazine compound represented by formula (II), a ketone compound represented by formula (III), a copper salt, and a halogen and add them to a solvent, and stir and react at 90 - 140 °C (preferably 120 - 140 °C, most preferably 140 °C) for 8 - 20 hours (preferably 8 - 13.5 hours, most preferably 13.5 hours). The obtained reaction solution is post-treated to obtain the N-triazine amide compound represented by formula (I); the molar ratio of the triazine compound represented by formula (II), the ketone compound represented by formula (III), the copper salt, and the halogen is 1:2.2 - 3.0:0.2 - 0.5:1.0 - 3.0 (preferably 1:2.2:0.4 - 0.5:1 - 1.5, particularly preferably 1:2.2:0.4:1.5); the solvent is one or a mixed solvent of two of N,N-dimethylformamide, chlorobenzene, 1,2-dichlorobenzene, and diethylene glycol dimethyl ether (preferably a mixed solvent of 1,2-dichlorobenzene and diethylene glycol dimethyl ether with a volume ratio of 2:1);

[0007]

[0008] In formula (I), (II), and (III), R 1 or R 2 are each independently a C1 - C10 alkyl group, or R 1 , R 2 and the N between the two combine to form a C4 - C8 heterocycle containing N or containing N and O; R3 is phenyl or phenyl substituted by C1-C10 alkyl, C1-C10 alkoxy or halogen.

[0009] Further, R 1 and R 2 are each independently preferably methyl, or R 1 and R 2 and the N therebetween combine to form a pyrrolidine ring, a piperidine ring or a morpholine ring.

[0010] Further, R 3 is phenyl or phenyl substituted by methyl, methoxy, halogen or nitro.

[0011] Furthermore, the N-triazine amide compound represented by formula (I) is one of the following:

[0012]

[0013] Preferably, the copper salt is one or a mixture of two or more of cuprous chloride, cuprous bromide, copper chloride, copper acetate, and is preferably copper chloride.

[0014] Preferably, the halogen in the reaction system is iodine.

[0015] Further, the volume of the solvent is 6-8 mL / mmol based on the amount of substance of the triazine compound represented by formula (II), and preferably 6 mL / mmol.

[0016] The present invention particularly recommends a method for preparing an N-triazine amide compound represented by formula (I), and the method is as follows:

[0017] Mix the triazine compound represented by formula (II), the ketone compound represented by formula (III), the copper salt and the halogen and add them to a solvent, stir and react at 140 °C for 13.5 hours, and post-treat the obtained reaction solution to obtain the N-triazine amide compound represented by formula (I); the molar ratio of the triazine compound represented by formula (II), the ketone compound represented by formula (III), the copper salt to the halogen is 1:2.2:0.4:1.5; the solvent is a mixed solvent of 1,2-dichlorobenzene and diethylene glycol dimethyl ether with a volume ratio of 2:1.

[0018] In the preparation method of the present invention, the post-treatment is: adding an aqueous sodium thiosulfate solution with a mass fraction of 10% to the reaction solution, extracting with ethyl acetate, combining the organic layers, drying with anhydrous sodium sulfate, concentrating, performing column chromatography separation with a mixed solution of petroleum ether and ethyl acetate with a volume ratio of 1:1 as the eluent, collecting the eluent containing the target compound, evaporating the solvent under reduced pressure and drying to obtain the N-triazine amide compound represented by formula (I).

[0019] Further, the volume of the sodium thiosulfate aqueous solution is 60 mL / mmol based on the amount of substance of the triazine compound shown in formula (II).

[0020] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0021] The present invention has developed a preparation method of N-triazine amide compounds. This process is convenient to operate and the raw materials are easily available, having certain application value. The N-triazine amide compounds provided by the present invention show certain anti-hepatocellular carcinoma and breast cancer biological activities, laying a foundation for new drug screening and development. (IV) Specific Embodiments

[0022] The present invention will be further described below through examples, but the protection scope of the present invention is not limited thereto.

[0023] The raw material triazine compound (II) used in the present invention is prepared by reacting an ester and biguanide under the action of sodium methoxide at room temperature; for the specific synthesis method, see the literature (Bioorganic & Medicinal Chemistry Letters, 19(2009), 5644 - 5647); the purity of the N-triazine amide compounds shown in formula (I) obtained in the examples is all above 97%.

[0024] Example 1: Preparation of Compound (I-1)

[0025] Add 2-amino-4-dimethylamino-1,3,5-triazine (69.6 mg, 0.5 mmol), acetophenone (132.2 mg, 1.10 mmol), I2 (190.4 mg, 0.75 mmol), CuCl2·2H2O (34.1 mg, 0.20 mmol) into a reaction vessel, and stir and react in a mixed solvent of 1,2-dichlorobenzene (2 mL) and diethylene glycol dimethyl ether (1 mL) in an oil bath at 140 °C for 13.5 hours, then stop the reaction. Add 30 ml of 10% sodium thiosulfate aqueous solution, extract with ethyl acetate (4×20 mL), combine the organic layers, dry over anhydrous sodium sulfate, filter, concentrate the filtrate, perform column chromatography (petroleum ether:ethyl acetate = 1:1, v:v), collect the eluate containing the target compound, and perform vacuum distillation and drying to obtain the target compound (I-1), 81 mg, with a yield of 67%.

[0026] 1 H NMR(500MHz,CDCl3)δ8.85(br,1H),8.32(s,1H),7.88(dt,J=7.5,1.0Hz,2H),7.57(tt,J=7.5,1.0Hz,1H),7.48(t,J=7.5Hz,2H),3.20(s,3H),3.17(s,3H).

[0027]

[0028] Example 2:

[0029] CuCl2·2H2O was changed to CuBr (28.6 mg, 0.2 mmol), and other operations were the same as in Example 1, 10.1 mg, and the yield was 8.1%.

[0030] Example 3:

[0031] CuCl2·2H2O was changed to Cu(OAc)2 (41.6 mg, 0.2 mmol), and other operations were the same as in Example 1, 9.0 mg, and the yield was 7.4%.

[0032] Example 4:

[0033] CuCl2·2H2O was changed to CuCl (19.8 mg, 0.2 mmol), and other operations were the same as in Example 1, 15.0 mg, and the yield was 12.3%.

[0034] Example 5:

[0035] The mixed solvent of 1,2-dichlorobenzene and diethylene glycol dimethyl ether was changed to chlorobenzene (3 mL), and other operations were the same as in Example 1, 17.9 mg, and the yield was 14.7%.

[0036] Example 6:

[0037] The mixed solvent of 1,2-dichlorobenzene and diethylene glycol dimethyl ether was changed to DMF (3 mL), and other operations were the same as in Example 1, 11.5 mg, and the yield was 9.5%.

[0038] Example 7:

[0039] The mixed solvent of 1,2-dichlorobenzene and diethylene glycol dimethyl ether was changed to diethylene glycol dimethyl ether (3 mL), and other operations were the same as in Example 1, 23 mg, and the yield was 19%.

[0040] Example 8:

[0041] The mixed solvent of 1,2-dichlorobenzene and diethylene glycol dimethyl ether was changed to 1,2-dichlorobenzene (3 mL), and other operations were the same as in Example 1, 42.3 mg, and the yield was 35%.

[0042] Example 9:

[0043] The mixed solvent of 1,2-dichlorobenzene and diethylene glycol dimethyl ether was changed to 1,2-dichlorobenzene (4 mL), and other operations were the same as in Example 1, 36.3 mg, and the yield was 30%.

[0044] Example 10:

[0045] The mixed solvent of 1,2-dichlorobenzene and diethylene glycol dimethyl ether was changed to 1,3,5-trichlorobenzene (4 mL), and other operations were the same as in Example 1, to obtain a trace amount of the target product.

[0046] Example 11:

[0047] The mixed solvent of 1,2-dichlorobenzene and diethylene glycol dimethyl ether was changed to dioxane (4 mL), and other operations were the same as in Example 1, to obtain a trace amount of the target product.

[0048] Example 12:

[0049] The amount of CuCl2·2H2O was increased to (42.6 mg, 0.25 mmol), and other operations were the same as in Example 1, 78.9 mg, and the yield was 63.3%.

[0050] Example 13:

[0051] The amount of CuCl2·2H2O was reduced to (17.1 mg, 0.1 mmol), and other operations were the same as in Example 1, 52 mg, and the yield was 43%.

[0052] Example 14:

[0053] The amount of I2 was reduced to (380.7 mg, 0.50 mmol), and other operations were the same as in Example 1, 68.8 mg, and the yield was 55.4%.

[0054] Example 15:

[0055] The amount of I2 was increased to (380.7 mg, 1.50 mmol), and other operations were the same as in Example 1, 11.3 mg, and the yield was 9.3%.

[0056] Example 16:

[0057] The amount of acetophenone was increased to (180.2 mg, 1.50 mmol), and other operations were the same as in Example 1, 70.3 mg, and the yield was 58.7%.

[0058] Example 17:

[0059] The reaction time was shortened to 8 hours, and other operations were the same as in Example 1, 40.5 mg, and the yield was 34.1%.

[0060] Example 18:

[0061] The reaction time was extended to 20 hours, and other operations were the same as in Example 1, 16 mg, and the yield was 13.3%.

[0062] Example 19:

[0063] The reaction temperature was changed to 90 °C, and other operations were the same as in Example 1. The yield was 3% with a product amount of 3.6 mg.

[0064] Example 20: Preparation of Compound (I-2)

[0065] The operation was the same as in Example 1, except that acetophenone was replaced with p-fluoroacetophenone (147.6 mg, 1.1 mmol). Other operations were the same as in Example 1. The target compound (I-2) was obtained, with a yield of 61% and a product amount of 77.6 mg.

[0066]

[0067] 1 H NMR (400 MHz, CDCl3) δ 8.80 (br, 1H), 8.35 (s, 1H), 7.96 - 7.87 (m, 2H), 7.21 - 7.12 (m, 2H), 3.20 (s, 3H), 3.17 (s, 3H).

[0068] Example 21: Preparation of Compound (I-3)

[0069] The operation was the same as in Example 1, except that acetophenone was replaced with p-chloroacetophenone (170.1 mg, 1.1 mmol). Other operations were the same as in Example 1. The target compound (I-3) was obtained, with a yield of 62% and a product amount of 82.5 mg.

[0070]

[0071] 1 H NMR (400 MHz, CDCl3) δ 8.78 (br, 1H), 8.37 (s, 1H), 7.85 - 7.81 (m, 2H), 7.48 - 7.44 (m, 2H), 3.21 (s, 3H), 3.17 (s, 3H).

[0072] Example 22: Preparation of Compound (I-4)

[0073] The operation was the same as in Example 1. Acetophenone was replaced with p-methylacetophenone (147.6 mg, 1.1 mmol). Other operations were the same as in Example 1. The target compound (I-4) was obtained, with a yield of 73.4% and a product amount of 92.7 mg.

[0074]

[0075] 11H NMR (400 MHz, CDCl3) δ 8.59 (br, 1H), 8.38 (s, 1H), 7.80 (d, J = 8.4 Hz, m, 2H), 7.28 (d, J = 8.4 Hz, 2H), 3.21 (s, 6H), 3.20 (s, 6H), 2.42 (s, 3H).

[0076] Example 23: Preparation of Compound (I-5)

[0077] The operation was the same as that in Example 1, except that acetophenone was replaced with 3-chloroacetophenone (170.1 mg, 1.1 mmol), and the other operations were the same as those in Example 1, to obtain the target compound (I-5), 80.0 mg, with a yield of 58%.

[0078]

[0079] 1 1H NMR (400 MHz, CDCl3) δ 9.13 (br, 1H), 8.40 (s, 1H), 7.93 (s, 1H), 7.80 (d, J = 7.8 Hz, 1H), 7.55 (d, J = 7.8 Hz, 1H), 7.42 (t, J = 7.8 Hz, 1H), 3.22 (s, 3H), 3.19 (s, 3H).

[0080] Example 24: Preparation of Compound (I-6)

[0081] The operation was the same as that in Example 1, except that 2-amino-4-dimethylamino-1,3,5-triazine was replaced with 2-amino-4-piperidin-1,3,5-triazine (89.6 mg, 0.50 mmol), and the other operations were the same as those in Example 1, to obtain the target compound (I-6), 105.8 mg, with a yield of 76%.

[0082]

[0083] 1 1H NMR (400 MHz, CDCl3) δ 8.85 (br, 1H), 8.49 - 8.18 (m, 1H), 7.88 (d, J = 7.8 Hz, 2H), 7.57 (t, J = 7.8 Hz, 1H), 7.48 (t, J = 7.8 Hz, 2H), 3.93 - 3.63 (m, 4H), 1.70 - 1.60 (m, 6H).

[0084] Example 25: Preparation of Compound (I-7)

[0085] The operation was the same as in Example 1, replacing 2-amino-4-dimethylamino-1,3,5-triazine with 2-amino-4-morpholino-1,3,5-triazine (90.6 mg, 0.50 mmol), and the other operations were the same as in Example 1 to obtain the target compound (I-7), 88.3 mg, with a yield of 60%.

[0086]

[0087] 1 H NMR(400MHz,CDCl3)δ8.76(br,1H),8.36 - 8.34(m,1H),7.90 - 7.87(m,2H),7.60 - 7.55(m,1H),7.53 - 7.45(m,2H),3.93 - 3.80(m,4H),3.78 - 3.70(m,4H)

[0088] Example 26: Preparation of compound (I-8)

[0089] The operation was the same as in Example 1, replacing acetophenone with o-fluoroacetophenone (179.6 mg, 1.3 mmol), and the other operations were the same as in Example 1 to obtain the target compound (I-8), 67.7 mg, with a yield of 53%.

[0090]

[0091] 1 H NMR(400MHz,CDCl3)δ9.01(br,1H),8.45(s,1H),8.10 - 8.05(m,1H),7.57 - 7.51(m,1H),7.33 - 7.27(m,1H),7.20 - 7.15(m,1H),3.21(s,3H),3.14(s,3H).

[0092] Example 27: Preparation of compound (I-9)

[0093] The operation was the same as in Example 1, replacing acetophenone with p-bromoacetophenone (218.9 mg, 1.1 mmol), and the other operations were the same as in Example 1 to obtain the target compound (I-9), 94.4 mg, with a yield of 60%.

[0094]

[0095] 1 H NMR(400MHz,CDCl3)δ9.04(br,1H),8.34(s,1H),7.75(d,J=8.6Hz,2H),7.61(d,J=8.6Hz,2H),3.20(s,3H),3.13(s,3H).

[0096] Example 28: Preparation of Compound (I-10)

[0097] The operation was the same as in Example 1, except that acetophenone was replaced with m-methylacetophenone (174.4 mg, 1.3 mmol), and the other operations were the same as in Example 1. The target compound (I-10), 64.0 mg, was obtained with a yield of 51%.

[0098]

[0099] 1 H NMR (400 MHz, CDCl3) δ 8.79 (br, 1H), 8.34 - 8.30 (m, 1H), 7.70 (s, 1H), 7.68 - 7.62 (m, 1H), 7.39 - 7.34 (m, 2H), 3.19 (s, 6H), 2.42 (s, 3H).

[0100] Example 29: Preparation of Compound (I-11)

[0101] The operation was the same as in Example 1, except that acetophenone was replaced with p-nitroacetophenone (214.7 mg, 1.3 mmol), and the other operations were the same as in Example 1. The target compound (I-11), 87.0 mg, was obtained with a yield of 61%.

[0102]

[0103] 1 H NMR (400 MHz, DMSO) δ 11.12 (br, 1H), 8.43 (s, 1H), 8.31 (d, J = 8.5 Hz, 2H), 8.05 (d, J = 8.5 Hz, 2H), 3.11 (s, 3H), 2.98 (s, 3H).

[0104] Example 30: Preparation of Compound (I-12)

[0105] The operation was the same as in Example 1, except that acetophenone was replaced with m-bromoacetophenone (258.8 mg, 1.3 mmol), and the other operations were the same as in Example 1. The target compound (I-12), 78.4 mg, was obtained with a yield of 51%.

[0106]

[0107] 11H NMR (400 MHz, CDCl3) δ 8.82 (br, 1H), 8.39 (s, 1H), 8.08 - 8.01 (m, 1H), 7.82 - 7.80 (m, 1H), 7.71 - 7.68 (m, 1H), 7.37 (t, J = 7.9 Hz, 1H), 3.21 (s, 3H), 3.16 (s, 3H).

[0108] Example 31: Preparation of Compound (I-13)

[0109] The operation was the same as that in Example 1, except that acetophenone was replaced with m-methoxyacetophenone (225.3 mg, 1.5 mmol), and other operations were the same as those in Example 1. The target compound (I-13), 76.1 mg, was obtained with a yield of 56%.

[0110]

[0111] 1 1H NMR (400 MHz, CDCl3) δ 8.74 (br, 1H), 8.35 (s, 1H), 7.45 - 7.41 (m, 2H), 7.38 (t, J = 7.9 Hz, 1H), 7.12 - 7.09 (m, 1H), 3.86 (s, 3H), 3.20 (s, 3H), 3.19 (s, 3H).

[0112] Example 32: Preparation of Compound (I-14)

[0113] The operation was the same as that in Example 1, except that 2-amino-4-dimethylamino-1,3,5-triazine was replaced with 2-amino-4-pyrrolidin-1,3,5-triazine (82.6 mg, 0.50 mmol), and acetophenone was replaced with p-fluoroacetophenone (179.6 mg, 1.3 mmol). Other operations were the same as those in Example 1. The target compound (I-14), 62.2 mg, was obtained with a yield of 44.5%.

[0114]

[0115] 1 1H NMR (400 MHz, CDCl3) δ 8.85 (br, 1H), 8.34 (s, 1H), 7.93 - 7.88 (m, 2H), 7.26 - 7.11 (m, 2H), 3.60 - 3.52 (m, 4H), 2.00 - 1.94 (m, 4H).

[0116] Example 33: Bioactivity Test against Human Hepatocellular Carcinoma Cells (HEPG2) or Human Breast Cancer Cells (T47D) In vitro bioactivity test method against human hepatocellular carcinoma cells (HEPG2) or human breast cancer cells (T47D): MTT method Experimental procedure:

[0117] 1) Preparation of samples: For soluble samples, dissolve 1 mg in 20 μL of DMSO. Take 2 μL and dilute it with 1000 μL of culture medium to a concentration of 100 μg / mL, and then further dilute it with culture medium to 60 μmol / L.

[0118] 2) Cell culture

[0119] 2.1) Preparation of culture medium: Each 1000 mL of culture medium contains 800,000 units of penicillin, 1.0 g of streptomycin, and 10% inactivated fetal bovine serum.

[0120] 2.2) Cell culture: Inoculate tumor cells into the culture medium and culture them in an incubator at 37°C with 5% CO2. Passage every 3 - 5 days.

[0121] 3) Determination of the inhibitory effect of samples on the growth of tumor cells

[0122] Digest the cells with EDTA - trypsin digestion solution and dilute them with culture medium to 1×10 5 / mL. Add 100 μL to each well of a 96 - well cell culture plate and culture it in an incubator at 37°C with 5% CO2. After 24 h of inoculation, add the sample diluted with culture medium, 100 μL to each well. Add 3 wells for each sample and culture it in an incubator at 37°C with 5% CO2. After 72 h, add 5 mg / mL of MTT, 10 μL to each well, and incubate at 37°C for 4 h. Then add 150 μL of DMSO to each well and shake with a shaker to completely dissolve the formazan. Measure the absorbance at a wavelength of 570 nm with an enzyme - linked immunosorbent assay (ELISA) reader. Use cells cultured in culture medium without samples and containing the same concentration of DMSO as a control under the same conditions, and calculate the inhibition rate of the samples on tumor cells. The results are shown in Table 1.

[0123] Using human hepatocellular carcinoma cells (HEPG2) or human breast cancer cells (T47D) as models, the inhibitory effects of 10 samples of compounds (I - 2) - (I - 11) on the growth of hepatocellular carcinoma cells or human breast cancer cells in vitro were determined (the results are shown in Table 1 for details).

[0124] Table 1. Inhibition rates of each compound on human hepatocellular carcinoma cells (HEPG2) or human breast cancer cells (T47D) (60 μmol / L)

[0125]

[0126]

Claims

1. A method for preparing an N-triazine amide compound represented by formula (I), characterized in that The method is as follows: The triazine compound shown in formula (II), the ketone compound shown in formula (III), a copper salt and iodine are mixed and added into a solvent, and stirred and reacted at 90 - 140 °C for 8 - 20 hours. The obtained reaction solution is post-treated to obtain the N-triazine amide compound shown in formula (I); the molar ratio of the triazine compound shown in formula (II), the ketone compound shown in formula (III), the copper salt to iodine is 1:2.2 - 3.0:0.2 - 0.5:1.0 - 3.0; the solvent is one or a mixed solvent of two of N,N-dimethylformamide, chlorobenzene, 1,2-dichlorobenzene, and diethylene glycol dimethyl ether; In formulas (I), (II), and (III), R 1 or R 2 is each independently preferably methyl, or R 1 , R 2 and the N therebetween combine to form a pyrrolidine ring, a piperidine ring or a morpholine ring; R 3 is phenyl or phenyl substituted by methyl, methoxy, halogen or nitro.

2. The preparation method of the N-triazine amide compound represented by formula (I) according to claim 1, characterized in that: The N-triazine amide compound shown in formula (I) is one of the following:

3. The preparation method of the N-triazine amide compound represented by formula (I) according to claim 1, characterized in that: The temperature of the stirring reaction is 120 - 140 °C.

4. The preparation method of the N-triazine amide compound shown by formula (I) as described in claim 1, characterized in that: The solvent is a mixed solvent of 1,2-dichlorobenzene and diethylene glycol dimethyl ether with a volume ratio of 2:

1.

5. The preparation method of the N-triazine amide compound shown in formula (I) as described in claim 1, characterized in that: The copper salt is one or a mixture of two or more of cuprous chloride, cuprous bromide, copper chloride, and copper acetate.

6. The preparation method of the N-triazine amide compound shown by formula (I) as described in claim 1, characterized in that: The volume of the solvent is 6 - 8 mL / mmol based on the amount of substance of the triazine compound shown in formula (II).

7. The preparation method of the N-triazine amide compound shown by formula (I) according to claim 1, characterized in that: The post-treatment is as follows: An aqueous sodium thiosulfate solution with a mass fraction of 10% is added to the reaction solution, extracted with ethyl acetate, the organic layers are combined, dried over anhydrous sodium sulfate, concentrated, and column chromatography separation is carried out using a mixed solution of petroleum ether and ethyl acetate with a volume ratio of 1:1 as the eluent. The eluate containing the target compound is collected, the solvent is removed by distillation under reduced pressure and dried to obtain the N-triazine amide compound shown in formula (I).

8. The preparation method of the N-triazine amide compound represented by formula (I) according to claim 7, characterized in that: The volume of the aqueous sodium thiosulfate solution is 60 mL / mmol based on the amount of substance of the triazine compound shown in formula (II).

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