CN106674136B - Pyrimidine antitumor compound and preparation method thereof - Google Patents

Abstract

The invention belongs to the field of medicine, in particular to a compound with a specific chemical structure of antitumor activity and a method for preparing the compound. These compounds belong to novel VEGFR-2 inhibitors, and show good inhibitory activity in the VEGFR-2 kinase inhibitory activity experiments in vitro. The compound can be used for preparing antitumor drugs.

Description

Pyrimidine antitumor compound and preparation method thereof

technical field

The invention belongs to the field of medicine, in particular to a compound with a specific chemical structure of antitumor activity and a method for preparing the compound.

Background technique

Cancer is a malignant tumor originating from epithelial tissue, characterized by rapid proliferation and metastasis of cells, and the mortality rate ranks first among all diseases. According to the latest global cancer data released by the International Agency for Research on Cancer, there were 14.1 million new cancer patients worldwide in 2012, and the annual growth rate will increase by 11%. Therefore, research on cancer treatment has been closely watched all over the world.

At present, there are four main ways of clinical treatment of cancer: surgery, radiation therapy, chemotherapy and immunotherapy. Compared with the other three methods, chemotherapy treatment is generally painless, and the killing power to cancer cells is difficult to achieve by other methods of treating cancer. But most chemical drugs are not specific. While killing cancer cells, they will cause damage to normal tissue cells around the tumor. These tissues usually repair themselves only after chemotherapy. Therefore, the search for anti-tumor drugs with strong targeting, significant effect and high safety has become one of the hotspots in the research of global pharmaceutical developers.

With the continuous development of tumor molecular biology technology, people's further understanding of the pathogenesis of malignant tumors, and the continuous disclosure of various carcinogenic pathways, a variety of molecular targeted therapy drugs have been developed. Molecular targeted therapy is aimed at target molecules and their related signaling pathways that play a key role in the occurrence, development and metastasis of tumors, interfering or blocking their efficacy, and achieving the purpose of inhibiting tumor growth and metastasis. The drug enters the body and binds to the carcinogenic site in a targeted manner, causing specific death of tumor cells without causing damage to normal tissue cells. Currently, drugs that have entered the clinical stage include small molecule epidermal growth factor receptor (EGFR) inhibitors, monoclonal antibodies targeting certain specific cell markers, anti-tumor angiogenesis drugs, and multi-targeted kinase inhibitors (multi-targeted tyrosine kinase inhibitors, TKIs) and so on. In conclusion, with the development of molecular biology, cancer therapy has entered the era of targeted therapy. Therefore, further research and development of new anti-cancer drugs with strong targeting, high efficiency and low toxicity has become an important direction of current anti-tumor drug research.

SUMMARY OF THE INVENTION

In view of the above-mentioned problems, the present invention provides pyrimidine antitumor compounds and a preparation method thereof. Such compounds belong to novel types of VEGFR-2 inhibitors, and exhibit good inhibition in vitro on VEGFR-2 kinase inhibitory activity experiments. active.

In order to achieve the above-mentioned purpose, the general structural formula of the pyrimidine antitumor compound provided by the present invention is the general formula I or the general formula II, which is specifically as follows:

.

Wherein: the position of the acylhydrazine group can be the 2-position, the 3-position or the 4-position; X is a hydrogen atom, a methyl group, a fluorine atom, a chlorine atom or a bromine atom; the position of the substituent on the aromatic ring can be the 2-position, the 3-position Or 4-position substitution, the substitution can be mono- or polysubstituted.

The general structural formula of the anti-tumor compound is preferably: .

wherein: X is H, Cl or CH ₃ .

The structure of the compound with the general formula I is selected from any one of the following, but is not limited to the following compounds, as long as the compound structural formula satisfies the general formula, it is within the scope of the present invention.

N' -Phenyl-2-(uracil-5-carboxamido)benzohydrazide.

N' -(2-Tolyl)-2-(uracil-5-carboxamido)benzohydrazide.

N' -(3-Tolyl)-2-(uracil-5-carboxamido)benzohydrazide.

N' -(3-Chlorophenyl)-2-(uracil-5-carboxamido)benzohydrazide.

N' -Phenyl-3-(uracil-5-carboxamido)benzohydrazide.

N' -(2-Tolyl)-3-(uracil-5-carboxamido)benzohydrazide.

N' -(3-Tolyl)-3-(uracil-5-carboxamido)benzohydrazide.

N' -(3-chlorophenyl)-3-(uracil-5-carboxamido)benzohydrazide.

N' -Phenyl-4-(uracil-5-carboxamido)benzohydrazide.

N' -(2-Tolyl)-4-(uracil-5-carboxamido)benzohydrazide.

N' -(3-Tolyl)-4-(uracil-5-carboxamido)benzohydrazide.

N' -(3-Chlorophenyl)-4-(uracil-5-carboxamido)benzohydrazide.

The general structural formula of the anti-tumor compound is preferably: .

wherein: X is H, Cl or CH ₃ .

The structure of the compound with the general formula II is selected from any one of the following, but not limited to the following compounds, as long as the compound structural formula satisfies the general formula IV, it is within the scope of the present invention.

N' -Phenyl-2-(uracil-6-carboxamido)benzohydrazide.

N' -(2-Tolyl)-2-(uracil-6-carboxamido)benzohydrazide.

N' -(3-Tolyl)-2-(uracil-6-carboxamido)benzohydrazide.

N'-(3-Chlorophenyl)-2-(uracil-6-carboxamido)benzohydrazide.

N'-phenyl-3-(uracil-6-carboxamido)benzohydrazide.

N' -(2-Tolyl)-3-(uracil-6-carboxamido)benzohydrazide.

N' -(3-Tolyl)-3-(uracil-6-carboxamido)benzohydrazide.

N' -(3-Chlorophenyl)-3-(uracil-6-carboxamido)benzohydrazide.

N' -Phenyl-4-(uracil-6-carboxamido)benzohydrazide.

N' -(2-Tolyl)-4-(uracil-6-carboxamido)benzohydrazide.

N' -(3-Tolyl)-4-(uracil-6-carboxamido)benzohydrazide.

N' -(3-Chlorophenyl)-4-(uracil-6-carboxamido)benzohydrazide.

In order to achieve the above object, the present invention also provides the above-mentioned preparation method for preparing the antitumor compound.

The preparation methods of the compounds represented by the general formulas I and II specifically include the following steps.

Step 1. Carboxylic acid uracil is halogenated to obtain uracil acid chloride.

In step 2, nitrobenzoic acid and substituted phenylhydrazine are subjected to dehydration condensation and reduction to obtain aminobenzoylarylhydrazine.

In step 3, the pyrimidine acid chloride obtained in step 1 and the aminobenzoyl aryl hydrazine obtained in step 2 are subjected to acylation reaction to obtain compounds represented by general formulas I and II.

The compounds can be used for preparing antitumor drugs.

Beneficial effects of the present invention.

When designing the compound, the present invention analyzes the characteristics of the ligand-receptor interaction in the structure of the small molecule inhibitor and the VEGFR-2 complex, extracts the key pharmacophore and performs virtual screening, and designs a new type of structure of VEGFR-2 inhibitors. Pharmacological studies show that the compounds of the present invention have certain inhibitory activity on human gastric cancer BGC-823 cells, human gastric cancer MGC-803 cells, human leukemia K562 cells and human leukemia HL60 cells.

Detailed ways

Example 1.

The preparation method of N' -phenyl-2-(uracil-5-carboxamido) benzohydrazide.

Step (1), preparation of N' -phenyl-2-nitro-benzoic hydrazide: add o-nitrobenzoic acid (4 g, 23.94 mmol), 1-hydroxybenzotriazole (3.4 g, 25.13 mmol), DMF 40 ml, EDCI (4.82 g, 25.13 mmol) was slowly added at 0°C, stirred for half an hour, moved to room temperature and reacted for 2 h to obtain an active ester solution; added to another 250 ml reaction flask Phenylhydrazine hydrochloride (25.13mmol), pyridine (3.98g, 50.26ml), DMF 30ml, stirred at room temperature for 30min, slowly added the above prepared active ester solution at 0 ℃, reacted for 2h, moved to room temperature and reacted for 6h After the reaction was completed, the reaction solution was poured into water, the crude product was precipitated, stood for 30 min, suction filtered to obtain a filter cake, washed with water, and dried at 40 °C overnight to obtain 4.20 g of white solid, yield: 68.21%.

Step (2), preparation of N' -phenyl-2-amino-benzohydrazide: add zinc powder (4.07g, 62.20mmol), ammonium chloride (2.5g, 46.65mmol), ethanol to a 250ml reaction flask 90ml, 60ml water, a small amount of glacial acetic acid, react at 50°C for 30min, activate zinc powder; let cool, move the reaction flask to a cold well, control the temperature below 0°C, slowly add N'-substituted phenyl-o/m/ p-nitrobenzoic hydrazide (15.55mmol), stirred for 10min, moved to room temperature for reaction for 4h, after the reaction was completed, let stand for half an hour; pour the reaction solution into 20% NaHCO ₃ solution, extract with ethyl acetate, spin The crude product was obtained from dry ethyl acetate, and the ethyl acetate was recrystallized to obtain 1.20 g of pure product with a yield of 33.96%.

Step (3), preparation of uracil-5-formyl chloride: add uracil-5-carboxylic acid (2g, 12.81mmol), thionyl chloride (1.73ml, 19.22mmol), toluene 30 to a 100 mL reaction flask mL, two drops of DMF, refluxed for 24 hours, cooled, and suction filtered to obtain uracil-5-carbonyl chloride, 2.12 g of white powder, and the yield was 94.80%.

Step (4), preparation of N' -phenyl-2-(uracil-5-carboxamido) benzoic hydrazide (A1): add N' -phenyl-2-amino-benzene into a 100ml reaction flask Formic hydrazide (2.86 mmol), DMF 20 ml, pyridine (0.45 g, 5.73 mmol), uracil-5-carbonyl chloride (0.5 g, 2.86 mmol) was slowly added at 0 °C, the reaction was carried out for 1 h, and the reaction was continued at room temperature for 12 h; After completion, the reaction solution was slowly poured into water, and a solid was precipitated. The pH value of the solution was adjusted to 2-3 with 10% dilute hydrochloric acid, and the solid was obtained by suction filtration, which was washed with saturated Na ₂ CO ₃ solution to obtain a white solid powder. Dry overnight to obtain 0.35 g of the target compound, yield: 33.44%. ¹ H NMR (600 MHz, DMSO) δ 11.63 (s, 1H), 10.27 (s, 1H), 8.34 – 8.19 (m, 2H), 7.86 (s, 1H), 7.71 (d, J = 7.2 Hz, 1H ), 7.50 (t, J = 7.5 Hz, 1H), 7.18 (dt, J = 15.6, 7.5 Hz, 3H), 6.83 (d, J = 7.8 Hz, 2H), 6.72 (t, J = 7.3 Hz, 1H) ). ¹³ CNMR (151 MHz, DMSO-d ₆ ) δ 167.6, 164.0, 161.9, 152.9, 152.1, 149.6, 137.4, 131.3, 129.1, 128.5, 125.0, 123.7, 123.7, 119.0, 112.7,

Example 2.

The preparation method of N' -(2-tolyl)-2-(uracil-5-carboxamido) benzohydrazide.

Using o-methylphenylhydrazine as raw material, according to the step (1) of Example 1 to synthesize N' -(2-methylphenyl)-2-nitro-benzohydrazide, and according to the step (4) of Example 1 to prepare A pale yellow solid was obtained, which was the target compound. ¹ H NMR (600 MHz, DMSO)δ 11.64 (s, 3H), 10.34 (s, 1H), 8.25 (d, J = 6.0 Hz, 2H), 7.75 (d, J = 7.5 Hz, 1H), 7.52 ( t, J = 7.7 Hz, 1H), 7.23 (dd, J = 16.6, 9.1 Hz, 2H), 7.02 (dd, J = 18.1, 7.5 Hz, 2H), 6.77 (d, J = 8.0 Hz, 1H), 6.68 (t, J = 7.3 Hz, 1H), 2.21 (s, 3H). ¹³ C NMR (151 MHz, DMSO-D ₆ ) Δ 167.5, 163.6, 161.2, 151.0, 149.3,146.8, 137.3, 131.5, 130.4, 128.8, 124.9, 123.8, 119.2, 1111.4, 104.8, 17.7.7.7.7.7.7.7.7.7.7.7.7.7. .

Example 3.

The preparation method of N' -(3-tolyl)-2-(uracil-5-carboxamido) benzohydrazide.

Using m-methylphenylhydrazine as raw material, N' -(3-methylphenyl)-2-nitro-benzohydrazide was synthesized according to step (1) of Example 1, and obtained according to step (4) of Example 1 The white solid is the target compound. ¹ H NMR (600 MHz, DMSO) δ11.63 (s, 2H), 10.27 (s, 1H), 8.24 (s, 2H), 7.76 (d, J = 40.7 Hz, 2H), 7.51(s, 1H) , 7.23 (s, 1H), 7.02 (s, 1H), 6.71 – 6.50 (m, 3H), 2.20 (s, 3H). ¹³ C NMR (151 MHz, DMSO-d ₆ ) d 167.5, 163.6, 161.3, 151.2, 149.6, 138.2, 137.3, 131.4, 129.0, 128.4, 124.9, 123.8, 119.9, 113.2, 110.7.0

Example 4.

The preparation method of N' -(3-chlorophenyl)-2-(uracil-5-carboxamido) benzohydrazide.

Using m-methylphenylhydrazine as raw material, N'-(3-methylphenyl)-2-nitro-benzohydrazide was synthesized according to step (1) of Example 1, and obtained according to step (4) of Example 1 The white solid is the target compound. ¹ H NMR (600 MHz, DMSO) δ11.59 (s, 3H), 10.34 (s, 1H), 8.23 (dd, J = 20.5, 11.5 Hz, 3H), 7.71 (d, J =7.2 Hz, 1H) , 7.52 (t, J = 7.6 Hz, 1H), 7.24 (t, J = 7.5 Hz, 1H), 7.16 (t, J =7.9 Hz, 1H), 6.86 – 6.68 (m, 3H). ¹³ C NMR (151 MHz, DMSO-d ₆ ) δ 167.6, 163.7, 161.3, 151.3, 151.1, 149.7, 137.1, 133.9, 131.5, 130.8, 128.4, 125.0, 124.0, 123.9, 11.3.

Example 5.

The preparation method of N' -phenyl-3-(uracil-5-carboxamido) benzohydrazide.

Using m-aminobenzoic acid as a raw material, according to the step (1) of Example 1 to synthesize N' -phenyl-3-nitro-benzohydrazide, and according to the step (4) of Example 1 to obtain 0.38 g of a white solid, namely As the target compound, the yield: 36.31%. ¹ H NMR (600MHz, DMSO) δ 11.89 (s, 2H), 11.07 (s, 1H), 10.39 (d, J = 2.7 Hz, 1H), 8.30 (s, 1H), 8.10 (s, 1H), 7.91 (d, J = 2.4 Hz, 2H), 7.65 (d, J = 7.7 Hz, 1H), 7.48 (t, J = 7.9 Hz, 1H), 7.16 (t, J = 7.8 Hz, 2H), 6.79 (d , J = 7.9 Hz, 2H), 6.72 (t, J = 7.3 Hz, 1H). ¹³ C NMR (151 MHz, DMSO-d ₆ ) δ 166.4, 165.0, 160.9, 150.8, 149.8, 149.3, 138.7, 134.3, 129.6, 129.1, 123.0, 122.8, 119.0, 119.0, 112.7, 119.0.

Example 6.

The preparation method of N' -(2-tolyl)-3-(uracil-5-carboxamido) benzohydrazide.

Using m-aminobenzoic acid and o-toluidine hydrazide as raw materials, according to the step (1) of Example 1, N' -(2-methylphenyl)-3-nitro-benzoic acid hydrazide was synthesized, and according to Example 1 In step (4), 0.42 g of yellow solid powder was obtained, which was the target compound, and the yield was 38.65%. ¹ H NMR (600 MHz, DMSO) δ 11.10 (s, 1H), 10.43 (dd, J = 18.5, 2.5 Hz, 1H), 8.30 (s, 1H), 8.12 (d, J = 11.2 Hz, 1H), 7.92 (dd, J = 8.1, 1.3Hz, 1H), 7.82 – 7.73 (m, 1H), 7.66 (d, J = 7.7 Hz, 1H), 7.50 (dt, J = 15.9,7.9 Hz, 1H), 7.29 (t, J = 3.2 Hz, 1H), 7.02 (dd, J = 12.1, 7.4 Hz, 2H), 6.73– 6.65 (m, 2H), 2.21 (s, 3H). ¹³ C NMR (151 MHz, DMSO-D ₆ ) Δ 166.4, 165.0, 161.0,151.1, 149.7, 147.1, 138.8, 134.3, 130.4, 129.6, 123.0, 122.3,119.0, 111.4, 104.1, 17.7.7.7.7.7.7.7.7 .

Example 7.

The preparation method of N' -(3-tolyl)-3-(uracil-5-carboxamido) benzohydrazide.

Using m-aminobenzoic acid and m-toluidine hydrazine as raw materials, according to the step (1) of Example 1 to synthesize N'-(3-methylphenyl)-3-nitro-benzoic acid hydrazide, and according to Example 1 In step (4), 0.46 g of yellow solid powder was obtained, which was the target compound, and the yield was 42.33%. ¹ H NMR (DMSO-d6 , 600MHz): δ 8.30 (br. s., 1H), 7.98 (d, J=7.3Hz, 1H), 7.83 (s, 1H), 7.65 (d, J=7.7 Hz, 1H), 7.47 (t, J=7.9 Hz, 1H), 7.03(t, J=7.6 Hz, 1H), 6.61 (br. s., 1H), 6.54 (d, J=7.3 Hz, 1H), 2.21 (s, 3H). ¹³ CNMR (151 MHz, DMSO-d ₆ ) δ 166.4, 149.9, 138.2, 134.3, 129.6, 129.0, 128.9, 123.0, 119.9, 119.0, 113.2, 113.2, 113.2, 110.0, 21.7.

Example 8.

The preparation method of N' -(3-chlorophenyl)-3-(uracil-5-carboxamido) benzohydrazide.

Using m-aminobenzoic acid and m-chlorophenylhydrazine as raw materials, according to the step (1) of Example 1, N'-(3-chlorophenyl)-3-nitro-benzohydrazide was synthesized, and according to the step (1) of Example 1 ( 4) 0.40 g of white solid powder was obtained, which was the target compound, and the yield was 34.93%. ¹ H NMR (600 MHz, DMSO) δ 11.33 (s, 1H), 10.44 (s, 1H), 8.39 (s, 1H), 8.23 (s, 1H), 8.09 (s, 1H), 7.92 (d, J = 8.0 Hz, 1H), 7.60 (d, J = 7.7 Hz, 1H), 7.46 (t, J = 7.9 Hz, 1H), 7.17 (t, J = 8.0 Hz, 1H), 6.75 (dd, J = 11.2 , 4.9 Hz, 3H). ¹³ C NMR (151 MHz, DMSO-d ₆ ) δ 166.6, 165.7, 162.4, 151.5, 139.4, 133.9, 133.9, 130.8, 129.6, 122.9, 122.2, 118.7, 118.4, 111.9, 111.3.

Example 9.

The preparation method of N' -phenyl-4-(uracil-5-carboxamido) benzohydrazide.

Using p-aminobenzoic acid raw material, according to the step (1) of Example 1 to synthesize N' -phenyl-4-nitro-benzohydrazide, and according to the step (4) of Example 1 to obtain the target compound 0.41g, the yield : 39.18%. ¹ H NMR (600 MHz, DMSO) δ 10.34 (s, 1H), 9.83 (s, 1H), 8.28 (s, 1H), 7.99 (dd, J = 18.2, 7.7 Hz, 1H), 7.90 (s, 1H) ), 7.86 (d, J = 7.2 Hz, 1H), 7.57 (t, J = 7.5 Hz, 1H), 7.21 (dt, J= 15.6, 7.5 Hz, 3H), 6.82 (d, J = 7.8 Hz, 2H) ), 6.50 (t, J = 7.3 Hz, 1H). ¹³ CNMR (151 MHz, DMSO-D ₆ ) Δ 166.1, 164.9, 161.0, 150.7, 150.0, 149.3, 141.4,129.2, 129.9, 128.8, 128.5, 123.2, 122.9.4, 112.8.112.7, 112.8, 112.7 104.2.

Example 10.

The preparation method of N' -(2-tolyl)-4-(uracil-5-carboxamido) benzohydrazide.

Using p-aminobenzoic acid and o-toluidine hydrazine as raw materials, according to the step (1) of Example 1, N' -(2-methylphenyl)-4-nitro-benzoic acid hydrazide was synthesized, and according to the step (1) of Example 1 ( 4) 0.41 g of yellow solid powder was obtained, which was the target compound, and the yield was 37.73%. ¹ H NMR (600 MHz, DMSO) δ 11.49 (s, 1H), 10.30 (s, 1H), 8.42 (d, J =8.3 Hz, 1H), 7.91 (d, J = 8.5 Hz, 2H), 7.74 ( d, J = 8.5 Hz, 2H), 7.22 (s, 1H), 7.02 (dd, J = 11.6, 7.5 Hz, 2H), 6.73 – 6.63 (m, 2H), 2.21 (s, 3H). ¹³ CNMR (151 MHz, DMSO-d ₆ ) δ 166.1, 147.3, 142.3, 130.3, 128.8, 127.5, 126.8, 122.3, 119.0, 111.5, 17.7. ¹³ C NMR (151 MHz, DMSO-D ₆ ) Δ 166.4, 165.0, 161.0,151.1, 149.7, 147.1, 138.8, 134.3, 130.4, 129.6, 123.0, 122.3,119.0, 111.4, 104.1, 17.7.7.7.7.7.7.7.7 .

Example 11.

The preparation method of N' -(3-methylphenyl)-4-(uracil-5-carboxamido) benzohydrazide.

Using p-aminobenzoic acid and m-toluidine hydrazine as raw materials, according to the step (1) of Example 1, N' -(3-methylphenyl)-4-nitro-benzohydrazide was synthesized, and according to the step (1) of Example 1 ( 4) 0.39 g of yellow solid powder was obtained, which was the target compound, and the yield was 35.89%. ¹ H NMR (600 MHz, DMSO) δ 11.75 (s, 1H), 10.22 (s, 1H), 8.50 (s, 1H), 7.88 (d, J = 8.6 Hz, 2H), 7.79 – 7.67 (m, 3H) ), 7.02 (t, J = 7.7 Hz, 1H), 6.63 – 6.56 (m, 2H), 6.53 (d, J = 7.4 Hz, 1H), 2.21 (s, 3H). ¹³ C NMR (151 MHz, DMSO-d ₆ ) δ 166.7, 166.3, 164.4, 163.2, 160.1, 150.1, 143.0, 138.1, 129.0, 128.7, 126.8, 119.8, 118.6, 113.3, 6.1, 2.9.

Example 12.

The preparation method of N' -(3-chlorophenyl)-4-(uracil-5-carboxamido) benzohydrazide.

Using p-aminobenzoic acid and m-chlorophenylhydrazine as raw materials, according to the step (1) of Example 1 to synthesize N' -(3-methylphenyl)-4-nitro-benzoic acid hydrazide, and according to the step (4) of Example 1 ) to obtain 0.38 g of yellow solid powder, which is the target compound, yield: 33.18%. ¹ H NMR (600 MHz, DMSO) δ 11.59 (s, 1H), 10.31 (s, 1H), 8.45 (s, 1H), 8.18 (s, 1H), 7.89 (d, J = 8.5 Hz, 2H), 7.74 (d, J = 8.5 Hz, 2H), 7.16 (t, J = 8.0 Hz, 1H), 6.83 – 6.67 (m, 3H). ¹³ C NMR (151 MHz, DMSO-d ₆ ) δ 166.3, 166.2, 163.4, 159.1, 157.3, 151.7, 142.7, 133.8, 130.8, 128.8, 126.9, 118.9, 118.3, 111.9, 111.3,

Example 13.

The preparation method of N' -phenyl-2-(uracil-6-carboxamido) benzohydrazide.

Using uracil-6-carboxylic acid as a raw material, according to the step (3) of Example 1 to synthesize uracil-6-carbonyl chloride, and according to the step (4) of Example 1 to obtain 0.42 g of white solid powder, which is the target compound, and received Rate: 40.13%. ¹ H NMR (600MHz, DMSO) δ 10.14 (d, J = 83.4 Hz, 1H), 8.52 (d, J = 8.2 Hz, 1H), 8.06 (s, 1H), 7.93 (d, J = 7.7 Hz, 1H) ), 7.56 (t, J = 7.5 Hz, 1H), 7.20 (dt, J = 41.3, 7.7 Hz, 4H), 6.87 (d, J = 7.8 Hz, 2H), 6.73 (t, J = 7.0 Hz, 1H) ), 5.92 (s,1H). ¹³ C NMR (151 MHz, DMSO-d ₆ ) δ 166.4, 166.2, 162.6, 151.6, 149.6, 137.4, 131.3, 129.1, 128.5, 125.0, 123.7, 123.7, 119.0, 112.7, 103.8.

Example 14.

The preparation method of N' -(2-tolyl)-2-(uracil-6-carboxamido) benzohydrazide.

Using o-methylphenylhydrazine as raw material, according to step (1) in Example 1, N' -(2-methylphenyl)-2-nitro-benzoic acid hydrazide was synthesized, and then uracil-6-carboxylic acid was used as raw material , according to the step (3) of Example 1 to synthesize uracil-6-formyl chloride, and finally according to the step (4) of Example 1 to obtain 0.44g of white solid powder, which is the target compound, the yield: 40.49%. ¹ H NMR (600MHz, DMSO) δ 10.09 (d, J = 27.4 Hz, 1H), 8.57 (d, J = 7.9 Hz, 1H), 7.95 (d, J = 7.4 Hz, 1H), 7.56 (s, 1H) ), 7.47 (s, 1H), 7.24 (t, J = 6.9 Hz, 1H), 7.05 (s, 2H), 6.86 (s, 1H), 6.74 – 6.65 (m, 1H), 5.90 (s, 1H) , 2.24 (s, 3H). ¹³ C NMR (151 MHz, DMSO-d ₆ ) δ 169.1, 166.9, 166.8, 166.8, 150.3, 146.6, 138.4, 132.5, 130.4, 128.9, 126.9, 123.6, 122.7, 1192.0, 1119.3

Example 15.

The preparation method of N' -(3-methylphenyl)-2-(uracil-6-carboxamido) benzohydrazide.

N' -(3-methylphenyl)-2-nitro-benzoic acid hydrazide was synthesized using m-methylphenylhydrazine as a raw material according to step (1) of Example 1, and then uracil-6-carboxylic acid was used as a raw material, Uracil-6-formyl chloride was synthesized according to the step (3) of Example 1, and finally 0.46 g of a white solid was obtained according to the step (4) of Example 1, which was the target compound, and the yield was 42.33%. ¹ H NMR (600 MHz, DMSO) δ 10.03 (s, 1H), 8.58 (d, J = 8.3 Hz, 1H), 8.03 – 7.86 (m, 2H), 7.56 (t, J = 7.2 Hz, 1H), 7.23 (t, J = 7.3 Hz, 1H), 7.05 (t, J = 7.6 Hz, 1H), 6.70(d, J = 7.9 Hz, 2H), 6.56 (d, J = 7.0 Hz, 1H), 5.88 ( s, 1H), 2.22 (s, 3H). ¹³ CNMR (151 MHz, DMSO-d ₆ ) δ 167.0, 149.4, 138.5, 138.3, 129.1, 128.8, 123.5, 120.8, 120.3, 113.6, 110.3, 96.3, 21.7.

Example 16.

The preparation method of N' -(3-chlorophenyl)-2-(uracil-6-carboxamido) benzohydrazide.

Use m-chlorophenylhydrazine as raw material to synthesize N' -(3-chlorophenyl)-2-nitro-benzoic acid hydrazide according to step (1) of Example 1, and then use uracil-6-carboxylic acid as raw material, according to the implementation of Example 1 Step (3) was used to synthesize uracil-6-formyl chloride, and finally 0.36 g of white solid was obtained according to step (4) of Example 1, which was the target compound, yield: 31.43%. ¹ H NMR (600 MHz, DMSO) δ10.11 (s, 1H), 8.09 (s, 1H), 7.64 (s, 2H), 7.16 (s, 2H), 6.73 (s, 4H), 6.55(s, 1H), 6.39 (s, 2H). ¹³ C NMR (151 MHz, DMSO-d ₆ ) δ 169.2, 151.8, 150.3, 133.8, 132.7, 130.8, 128.3, 118.3, 116.8, 115.1, 112.8, 111.8, 111.2.

Example 17.

The preparation method of N' -phenyl-3-(uracil-6-carboxamido) benzohydrazide.

Using m-aminobenzoic acid as the raw material, according to the step (1) of Example 1, N' -phenyl-3-nitro-benzohydrazide was synthesized, and then using uracil-6-carboxylic acid as the raw material, according to the step (3) of Example 1 ) to synthesize uracil-6-formyl chloride, and finally, according to step (4) of Example 1, 0.39 g of white solid powder was obtained, which was the target compound, and the yield was 37.26%. ¹ H NMR (600 MHz, DMSO)δ 10.35 (s, 1H), 9.84 (s, 1H), 8.29 (s, 1H), 8.04 – 7.87 (m, 2H), 7.64 (d, J = 7.6 Hz, 1H ), 7.48 (t, J = 7.9 Hz, 1H), 7.16 (t, J = 7.8 Hz, 2H), 6.86 –6.65 (m, 3H), 5.87 (s, 1H). ¹³ C NMR (151 MHz, DMSO-d ₆ ) δ 167.4, 166.7, 163.8, 159.9, 149.8, 138.7, 134.2, 129.3, 129.1, 123.0, 122.7, 119.2, 119.0, 112.7, 95.4.

Example 18.

The preparation method of N' -(2-tolyl)-3-(uracil-6-carboxamido) benzohydrazide.

Using m-aminobenzoic acid and o-toluidine hydrazine as raw materials, according to the step (1) of Example 1, N' -(2-methylphenyl)-3-nitro-benzoic acid hydrazide was synthesized, and then uracil-6-carboxylic acid was used. As raw material, uracil-6-formyl chloride was synthesized according to step (3) of Example 1, and finally 0.33 g of white solid powder was obtained according to step (4) of Example 1, which was the target compound, yield: 30.37%. ¹ HNMR (DMSO-d6 , 600MHz): δ 8.30 (s, 1H), 8.08 - 8.14 (m, 1H), 7.88 - 7.98 (m, 1H), 7.65 (d, J=7.7 Hz, 1H), 7.46- 7.53 (m, 1H), 7.28-7.30 (m, 1H), 7.03 (d, J=7.2 Hz, 1H), 6.70 (d, J=8.1 Hz, 1H), 2.18 - 2.22 (m, 4H). ¹³ C NMR (151 MHz, DMSO-D ₆ ) Δ 166.1, 151.4, 151.2, 148.1, 143.8, 134.2, 134.0, 133.9, 130.9, 130.9, 129.7, 121.6, 118.5, 111.9, 111.9, 111.9, 111.9 , 111.3.

Example 19.

The preparation method of N' -(3-methylphenyl)-3-(uracil-6-carboxamido) benzohydrazide.

Using m-aminobenzoic acid and m-toluidine hydrazine as raw materials, according to the step (1) of Example 1, N' -(3-methylphenyl)-3-nitro-benzoic acid hydrazide was synthesized, and then uracil-6-carboxylic acid was used As raw material, uracil-6-formyl chloride was synthesized according to step (3) of Example 1, and finally 0.35 g of white solid powder was obtained according to step (4) of Example 1, which was the target compound, yield: 33.21%. ¹ HNMR (DMSO-d6 , 600MHz): δ 8.31 (d, J=7.7 Hz, 1H), 8.07 (s, 1H), 7.85 - 7.94(m, 1H), 7.75 - 7.84 (m, 1H), 7.66 - 7.74 (m, 1H), 7.62 (d, J=7.3 Hz, 1H), 7.46 (t, J=7.9 Hz, 1H), 7.03 - 7.13 (m, 1H), 6.60 - 6.66 (m, 1H), 2.14 - 2.24(m, 4H). ¹³ C NMR (151 MHz, DMSO-d ₆ ) δ 166.0, 165.9, 161.7, 147.3, 138.2, 134.3, 129.6, 129.0, 128.9, 123.0, 119.9, 119.0, 113.2, 113.2, 113.2, 113.2, 113.2

Example 20.

The preparation method of N' -(3-chlorophenyl)-3-(uracil-6-carboxamido) benzohydrazide.

Use m-aminobenzoic acid and m-chlorophenylhydrazine as raw materials to synthesize N' -(3-chlorophenyl)-3-nitro-benzoic acid hydrazide according to the step (1) of Example 1, and then use uracil-6-carboxylic acid As raw material, uracil-6-formyl chloride was synthesized according to step (3) of Example 1, and finally 0.42 g of white solid powder was obtained according to step (4) of Example 1, which was the target compound, yield: 36.67%. ¹ HNMR (DMSO-d6 , 600MHz): δ 11.32 (s, 1H), 10.43 (br. s., 1H), 8.39 (s, 1H), 8.19 - 8.27 (m, 1H), 8.08 (s, 1H) , 7.91 (d, J=7.9 Hz, 1H), 7.59 (d, J=7.5 Hz, 1H), 7.45 (t, J=7.9 Hz, 1H), 7.16 (t, J=8.0 Hz, 1H), 6.68 - 6.81 (m, 3H). ¹³ CNMR (151 MHz, DMSO-D ₆ ) Δ 166.1, 151.4, 151.2, 148.1, 143.8, 134.2, 134.0,133.9, 133.9, 130.9, 129.7, 121.6, 118.5, 111.9,111.4, 111.9,111.4 111.3.

Example 21.

The preparation method of N' -phenyl-4-(uracil-6-carboxamido) benzohydrazide.

Using p-aminobenzoic acid as raw material to synthesize N' -phenyl-3-nitro-benzohydrazide according to the steps of Example 1, and then using uracil-6-carboxylic acid as the raw material, according to the steps (3) of Example 1 to synthesize urine Pyrimidine-6-formyl chloride, finally according to the step (4) of Example 1 to obtain 0.38 g of white solid powder, which is the target compound, yield: 36.31%. ¹ H NMR (DMSO-d6 , 600MHz): δ 8.40 (d, J=8.7 Hz, 1H), 8.26 - 8.32 (m, 1H), 8.18 (s, 1H), 8.15(d, J=2.3 Hz, 1H) ), 8.10 (d, J=8.5 Hz, 1H), 8.01 - 8.06 (m, 1H), 7.97 (dd, J=13.0, 2.6 Hz, 1H), 7.86 (d, J=2.6 Hz, 1H). ¹³ C NMR (151 MHz, DMSO-d ₆ ) δ 150.0, 149.8, 149.7, 149.6, 129.2, 129.1, 128.9, 128.6, 128.5, 125.3, 123.2, 122.9, 119.6, 119.2, 112.8, 119.2, 112.8, 119.2, 112.8, 119.0

Example 22.

The preparation method of N' -(2-tolyl)-4-(uracil-6-carboxamido) benzoic hydrazide.

Using p-aminobenzoic acid and o-toluidine as raw materials, according to the step (1) of Example 1, N' -(2-methylphenyl)-3-nitro-benzoic acid hydrazide was synthesized, and then uracil-6-carboxylic acid was used As raw material, uracil-6-formyl chloride was synthesized according to step (3) of Example 1, and finally 0.38 g of white solid powder was obtained according to step (4) of Example 1, which was the target compound, yield: 34.79%. ¹ HNMR (DMSO-d6 , 600MHz): δ 7.85 - 7.95 (m, J=8.5 Hz, 2H), 7.70 - 7.77 (m, J=8.5Hz, 2H), 7.21 (br. s., 1H), 6.96 - 7.05 (m, 2H), 6.58 - 6.76 (m, 2H), 2.11 -2.24 (m, 3H). ¹³ C NMR (151 MHz, DMSO-d ₆ ) δ 166.0, 165.9, 161.7, 147.3, 141.3, 130.3, 128.8, 128.6, 126.8, 122.3, 119.8, 119.1, 111.5, 98.5, 17.7.

Example 23.

The preparation method of N' -(3-tolyl)-4-(uracil-6-carboxamido) benzohydrazide.

Using p-aminobenzoic acid and m-toluidine hydrazine as raw materials, according to the step (1) of Example 1, N' -(3-methylphenyl)-3-nitro-benzoic acid hydrazide was synthesized, and then uracil-6-carboxylic acid was used. As the raw material, uracil-6-formyl chloride was synthesized according to the step (3) of Example 1, and finally 0.44 g of white solid powder was obtained according to the step (4) of Example 1, which was the target compound, and the yield was 40.49%. ¹ HNMR (600 MHz, DMSO) δ 9.86 (d, J = 2.5 Hz, 1H), 7.64 (s, 1H), 7.63 (s, 1H), 7.60 (d, J = 2.5 Hz, 1H), 7.00 (t , J = 7.7 Hz, 1H), 6.60 – 6.48 (m, 6H), 5.68(s, 2H), 2.19 (s, 3H). ¹³ C NMR (151 MHz, DMSO-d ₆ ) d 166.7, 152.4, 150.5, 139.7, 138.0, 134.1, 133.0, 129.9, 128.9, 123.0, 121.0, 119.8, 119.6, 113.4, 1.703.2

Example 24.

The preparation method of N' -(3-chlorophenyl)-4-(uracil-6-carboxamido) benzohydrazide.

Using p-aminobenzoic acid and m-chlorophenylhydrazine as raw materials, according to the step (1) of Example 1, N' -(3-chlorophenyl)-3-nitro-benzoic acid hydrazide was synthesized, and then uracil-6-carboxylic acid was used As raw material, uracil-6-formyl chloride was synthesized according to step (3) of Example 1, and finally 0.40 g of white solid powder was obtained according to step (4) of Example 1, which was the target compound, yield: 34.93%. ¹ HNMR (600 MHz, DMSO) δ 10.36 (s, 3H), 8.20 (s, 1H), 7.91 (dd, J = 26.9, 8.5Hz, 4H), 7.16 (t, J = 8.0 Hz, 1H), 6.83 – 6.68 (m, 3H), 5.99 (s, 1H). ¹³ C NMR (151 MHz, DMSO-d ₆ ) d 166.4, 166.2, 162.6, 151.6, 141.5, 133.8, 130.8, 128.6, 128.2, 119.7, 118.4, 111.9, 111.3, 97.4.

1. In vitro VEGFR-2 kinase inhibitory activity assay.

Using the Mobility Shift Assay method, in the case of Km ATP, the in vitro kinase VEGFR-2 was screened for compounds. The compound staurosporine was used as a standard control, and each compound was diluted to 10 concentration points for single-well detection.

1. Dilution of the compound.

Add 30 pL of 10 mM compounds of the present invention to EP tubes, and then add 7 μL of 100% DMSO to make up 100 μL of 2.5 mM compounds of the present invention. The compounds in the EP tube were transferred to the second column of the 96-well plate, and 20 μL was taken from it and added to the subsequent wells. At the same time, 60 μL of 100% DMSO was added, and 10 concentrations were sequentially diluted according to this ratio. The compound concentration range was as follows: 2.5 mM to 9.5 nM. In the first and last two wells of each row of the 96-well plate, 60 μL of 100% DMSO was added as a blank control. Pipette 5 μL from each well of the above 96-well plate into another 96-well plate, and add 45 μL of ultrapure water. Then transfer 5 μL from each well to the 384-well plate, that is, the solution in A1 of the 96-well plate is transferred to two wells A1 and A2 of the 384-well plate, and the solution in A2 (the highest concentration of the compound) is transferred to A3 and A4 of the 384-well plate, and so on. Therefore, 5 μL of 10% DMSO dissolves 5x compound in a 384-well reaction plate.

2. Kinase reaction.

Add kinase to 1x kinase buffer to form 2.5x enzyme solution; add FAM-labeled polypeptides and ATP to 1x kinase buffer to form 2.5x substrate solution; add 5x compound in 5 μL of 10% DMSO dissolved Add 10 μL of 2.5 times enzyme solution to the 384-well reaction plate; then incubate at room temperature for 10 minutes; then add 10 μL of 2.5 times substrate solution to the 384-well reaction plate; after incubation at 28°C for a certain period of time, add 25 μL of stop solution Terminate the reaction.

3. Inhibition rate calculation.

Read conversion rate data from Caliper and convert conversion rate into inhibition rate data.

Percent inhibition = (max-conversion)/(max-min)*100, max refers to the conversion rate of the DMSO control, and min refers to the conversion rate of the non-enzymatic control.

_IC50 data were obtained after XLfit.

The equation is as follows.

.

The experimental results are shown in Table 1.

Table 1 IC50 values of target compounds for VEGFR-2 kinase inhibitory activity.

A1-12 is the compound prepared in Example 1-12; B1-12 is the compound prepared in Example 13-24; the above experimental data show that the compound in the present invention has better VEGFR-2 kinase inhibitory activity, so it is In-depth research and development of new anti-tumor drugs have opened up new avenues.

Claims (7)

1. Pyrimidine anti-tumor compounds, characterized in that the general structural formula of the compound is general formula I or general formula II, specifically as follows, the position of the acylhydrazine group in the general formula can be 2-position, 3-position Position or position 4; X is a hydrogen atom, methyl group, fluorine atom, chlorine atom or bromine atom; the position of the X substituent can be substituted at the 2-position, 3-position or 4-position of the benzene ring, and the substitution can be mono-substituted or multi-substituted . 2. The pyrimidine antitumor compound according to claim 1, wherein the general formula of the antitumor compound is preferably: wherein: X is H, Cl or CH ₃ . 3. The pyrimidine antitumor compound according to claim 2, wherein the compound with the general formula I is: N' -phenyl-2-(uracil-5-carboxamido)benzyl Hydrazide; N' -(2-Tolyl)-2-(uracil-5-carboxamido)benzohydrazide; N' -(3-Tolyl)-2-(uracil-5-carboxamide N' -(3-chlorophenyl)-2-(uracil-5-carboxamido) benzoic hydrazide; N' -phenyl-3-(uracil-5-methyl) amido) benzoic hydrazide; N' -(2-tolyl)-3-(uracil-5-carboxamido) benzoic hydrazide; N' -(3-tolyl)-3-(uracil -5-Carboxamido)benzohydrazide; N' -(3-chlorophenyl)-3-(uracil-5-carboxamido)benzohydrazide; N' -phenyl-4-(urea Pyrimidine-5-carboxamido)benzohydrazide; N' -(2-tolyl)-4-(uracil-5-carboxamido)benzohydrazide; N' -(3-tolyl)- 4-(uracil-5-carboxamido)benzohydrazide; N' -(3-chlorophenyl)-4-(uracil-5-carboxamido)benzohydrazide. 4. The pyrimidine antitumor compound according to claim 1, wherein the general formula of the antitumor compound is preferably: wherein: X is H, Cl or CH ₃ . 5. The pyrimidine antitumor compound according to claim 4, wherein the compound of the general formula II is: N' -phenyl-2-(uracil-6-carboxamido)benzyl Hydrazide; N' -(2-Tolyl)-2-(uracil-6-carboxamido)benzohydrazide; N' -(3-Tolyl)-2-(uracil-6-carboxamide N'-(3-chlorophenyl)-2-(uracil-6-carboxamido)benzohydrazide;N'-phenyl-3-(uracil-6-methyl)amido)benzohydrazide;N'-(2-methylphenyl)-3-(uracil-6-carboxamido)benzohydrazide;N' -(3-methylphenyl)-3-(uracil -6-Carboxamido)benzohydrazide; N' -(3-chlorophenyl)-3-(uracil-6-carboxamido)benzohydrazide; N' -phenyl-4-(urea Pyrimidine-6-carboxamido)benzohydrazide; N' -(2-tolyl)-4-(uracil-6-carboxamido)benzohydrazide; N' -(3-tolyl)- 4-(uracil-6-carboxamido)benzohydrazide; N' -(3-chlorophenyl)-4-(uracil-6-carboxamido)benzohydrazide. 6. the preparation method of the antitumor compound as described in any one of claim 1-5, is characterized in that, specifically comprises the following steps: Step 1. Carboxylic acid uracil obtains uracil acid chloride by halogenation; Step 2, nitrobenzoic acid and substituted phenylhydrazine are subjected to dehydration condensation and reduction to obtain substituted aminobenzoylphenylhydrazine; Step 3. The uracil chloride obtained in step 1 and the substituted aminobenzoyl phenylhydrazine obtained in step 2 are subjected to acylation reaction to obtain compounds represented by general formulas I and II. 7. The compound according to any one of claims 1-5 is used for the preparation of anti-tumor drugs, the mechanism of which is to inhibit the activity of VEGFR-2 kinase.