CN104163818A - 2-amino oxazole compound and preparation method and application thereof - Google Patents

Abstract

The invention belongs to the field of pharmaceutical compounds synthesis, and relates to a Src / Abl kinase dual inhibitor drug 2-amino oxazole compound and a preparation method and application thereof. The invention provides a new drug with a broad market prospect.

Description

2-aminooxazole compounds and their preparation methods and uses

technical field

The invention belongs to the field of chemical synthesis, and in particular relates to the synthesis of Src/Abl kinase dual inhibitor drugs.

Background technique

Chronic myeloid leukemia (CML) is the first malignant tumor found to have fixed cytogenetic abnormalities. The global annual incidence is 1.6-2/100,000 people. Its main cause is the mutual translocation of chromosomes 9 and 22 . Afterwards, the oncogene Abl is transferred to the breakpoint region of the long arm to form the Bcr-Abl fusion gene, one of its products is an abnormal tyrosine kinase (tyrosine protein kinase, TPK) receptor protein P210 ^Bcr-Abl , which does not It needs to be combined with the corresponding ligand to be able to phosphorylate itself, and at the same time cause phosphorylation of many important substrate proteins, thereby activating multiple signal (including Src family members) transduction pathways, resulting in excessive cell proliferation, abnormal differentiation and apoptosis stymied. The current research agrees that the abnormally increased TPK activity of Bcr-Abl protein is the main reason for the occurrence of CML.

Contents of the invention

The object of the present invention is to provide a Src/Abl kinase dual inhibitor drug 2-aminooxazole compound and its preparation method and application.

The 2-aminooxazole compound structural formula is:

,

Wherein R=H, C _1-5 alkyl, halogen; R ₁ =H, C _1-5 alkyl, halogen, CN; X=N, CH, R ₃ =H, CH ₃ , halogen, CN, NO ₂ ; R ₄ =H, CH ₃ , halogen, CN, NO ₂ ;

_R2 is hydroxyethylpiperazine, hydroxyethylpiperidine, piperazineethylamino, morpholinoethylamino, piperidineethylamino

Piperazinemethylamino, morpholinomethylamino, piperidinylmethylamino, hydroxyethylpiperazinemethylamino, hydroxyethylpiperidinylmethylamino, C _1-5 alkylamino.

The present invention also provides a preparation method for the aforementioned 2-aminooxazole compounds, as shown in Figure 1 of the technical roadmap:

Including the following steps:

Substituted arylamine reacts with E-3-ethoxyacryloyl chloride in pyridine and tetrahydrofuran to obtain EN-aryl-3-ethoxyacrylamide; EN-aryl-3-ethoxyacrylamide reacts with NBS After the reaction, immediately condense and close the ring with urea to obtain the compound formula (4 a-c ) ; compound formula (4 a-c ) can obtain the final target product through inverse nucleophilic substitution with dichloropyrimidine or dichloropyridine and hydroxyethylpiperazine respectively; or compound formula (4 a-c ) can be obtained by reacting with aroyl chloride or alkyl Acyl chloride reaction to obtain the final target product.

The invention has the beneficial effects of providing a novel Src/Abl kinase dual inhibitor and a preparation method thereof, which has a high inhibitory rate and good effect on inhibiting K562 cell activity.

Description of drawings

Figure 1 is a synthetic route diagram of 2-aminooxazole compounds of the present invention.

Detailed ways

Below, the present invention will be further described according to the synthetic route diagram of Fig. 1 .

Example 1 Preparation of 2-aminooxazole compounds

EN-p-tolyl-3-ethoxyacrylamide (3a): Take p-toluidine (3.0g, 28mmol), pyridine (3.2mL, 40mmol) and tetrahydrofuran (40mL) into the reaction flask, ice bath, add Slowly add E-3-ethoxyacryloyl chloride ( 2 ) (3.6mL, 30mmol) dropwise, and react at room temperature for 2h;

Add 1mol/L HCl (7.0mL) at 0-5 ℃, then add 40mL water to it, concentrate under reduced pressure, add 30mL toluene to the concentrated solution, stir in ice bath for 10min, filter, wash the filter residue with 20mL water, vacuum Dry to obtain 5.0 g of pale white product, yield 87.0%, ¹ H-NMR (600 MHz, DMSO-d6) δ : 1.27 (t,3H, J =7.0Hz), 2.16 ( s, 3H ), 3.94 (q, 2H, J=7.0 Hz), 5.58 (d, 1H, J=12.4 Hz), 7.18 -7.23 (m, 2H, J=7.5Hz), 7.32(d, 1H, J=7.5 Hz), 7.46 (d, 1H, J=12.4 Hz), 9.29 (s, 1H) . IR (KBrcm-1): 3257, 1662, 1622, 1508, 1292, 1161;

2-Amino-N-p-tolyl-5-oxazole carboxamide (5a): In a 150mL three-necked flask, add 40mL of water, 40mL of dioxane and EN-p-tolyl-3-ethoxyacrylamide ( 3a ) (5.0g, 25mmol), ice bath, add NBS (4.4g, 25mmol) to the reaction bottle, keep at this temperature for 1h, then react at room temperature for 2h;

Add urea (1.7g, 29mmol) to the reaction bottle, react at room temperature for 0.5h, then react at 60°C for 3h, and react at 80°C for 1h, cool to room temperature, add concentrated ammonia water to adjust the pH to 8-9, concentrate, filter, and wash the filter cake with water , dried in vacuo to obtain 2.5 g of yellow product, yield 45.7%, mp271-272 °C. ¹ H-NMR (600 MHz, DMSO-d6 ) δ: 2.26(s, 3H ), 7.12( d, 2H, J=8.4 Hz), 7.30( s, 2H), 7.53 ( d, 2H, J=8.4 H z), 7.61(s, 1H ), 9.70 (s, 1H ), ¹³ C-NMR (DMSO-d6, 400 MHz) δ(ppm): 20.97, 42.11, 120.76, 120.76, 127.41, 128.50, 128.96, 128.96, 129.59, 129.59, 129.80, 129.80 , 133.22, 135.14, 136.59, 140.73, 159.74, 161.43, 170.28; Mass Spectra ( m/z ) [M + H] ⁺ : 217.49; , 1633, 1607, 1574, 1529, 1286, 1173;

N-p-tolyl-2-benzamide-5-oxazolecarboxamide (6d): take benzoic acid and 5mL dichloromethane and place in a 50mL single-necked bottle, ice-bath, drop (0.5mL, 6.95mmol ) thionyl chloride, and then slowly raise the temperature to 75°C until no gas escapes, and remove the solvent and excess thionyl chloride under reduced pressure. Add 5mL tetrahydrofuran to it, and transfer to the dropping funnel;

In a 50mL single-necked bottle, add 2-amino-N-p-tolyl-5-thiazole carboxamide ( 4a ) (0.5g, 2.15mmol), 5mL tetrahydrofuran and (0.7mL, 4.99mmol) triethylamine, ice bath, Slowly add the previously prepared benzoyl chloride dropwise, and continue to react for 1 hour, and then react at room temperature for 2 hours. The solvent was evaporated under reduced pressure, 5 mL of dichloromethane and 20 mL of water were added thereto, stirred for 5 min, filtered with suction, the filter residue was washed with 1 mL of dichloromethane and 5 mL of water, and recrystallized from acetone to obtain 0.62 g of a white solid with a yield of 86.1%, ¹ H-NMR (600 MHz, DMSO-d6 ) δ: 2.29(s, 3H ), 7.17( d, 2H, J=8.4H z), 7.57(t, 2H, J=7.8H z), 7.61( d, 2H, J=8.4H z), 7.67(t, 1H, J=7.2H z), 8.13(d, 2H, J=7.2H z), 8.40( s, 1H), 10.17(s, 1H ), 12.93 (s, 1H ) ， ¹³ C-NMR (DMSO-d6, 400 MHz) δ(ppm): 20.97, 42.11, 120.76, 120.76, 127.41, 128.50, 128.96, 128.96, 129.59, 129.59, 129.83.0, 24, 129 , 136.59, 140.73, 159.74, 161.43, 170.28; Mass Spectra( m/z ) [M + H] ⁺ 337.87, [M + Na] ⁺ 359.9; IR(KBrcm-1): 3454, 3136, 1676, 1634, 1514 , 1298, 1182.

Example 2

The preparation of 4a-c compounds is shown in Example 1;

Synthesis of 2-(6-chloro-2-methylpyrimidine-4-amino)-N-substituted phenyl-5-oxazolecarboxamide

Compound 4a-c (41 mmol), 4,6-dichloro-2-methylpyridine (8.0 g, 41 mmol) and THF (105 ml) were added to a 250 mL three-necked flask, cooled to -10- At ~5°C, freshly prepared sodium tert-butoxide (25.8 g, 250 mmol) was added, and the reaction was stirred at -10°C for 1.5 h. Add 2 mol/L hydrochloric acid to adjust to neutral, stir at 0°C for 1.5h, filter, wash the filter cake with a small amount of water, and dry to obtain light yellow solid 6 (15g, 88%) mp>300°C;

4 Synthesis of N-substituted phenyl-2-[6-[4-(3-hydroxyethyl)-1-piperazinyl]-2-methyl-4-pyrimidinylamino]-5-oxazolecarboxamide

The above compound (8.16 mmol) was dissolved in 30 mL N,N-dimethylformamide, 1-(2-hydroxyethyl)piperazine (3.2 g, 21 mmol) and triethylamine (1.6 g, 16 mmol), heated to 80 °C and stirred for 4 h. Cool to room temperature, slowly pour it into 100 mL of water, stir for 10 min, filter with suction, add the filter cake to 20 mL of ethanol aqueous solution with a volume fraction of 80%, heat to 80 °C, stir for 5 min, and then slowly add 5mL of water, cooled to room temperature, suction filtered, and dried to obtain 3.5 g of white solid, yield 85%.

Example 3 2-aminooxazole compounds of the present invention are tested for their inhibitory activity on human CML cells K562 cells

The method is described as follows: use the 2-aminooxazole compound of the present invention in K562 cells at different times, and use the MTT method to measure the concentration IC ₅₀ that causes 50% of uninfected cells to undergo cytopathic changes;

Materials and methods: The inhibitory activity of each compound on K562 cells was monitored by the inhibitory effect on drug-induced cytopathic changes;

The specific operation is as follows: in a CO ₂ incubator, the cells were cultured and passaged in RPMI1640 medium containing 10% inactivated calf serum. Centrifuge the cells in the logarithmic growth phase to make a 1* ¹⁰⁵ suspension, inoculate in RPMI-1640 containing 10% newborn bovine serum, inoculate 10,000 cells/well in a 96-well plate, add 100ul containing Add 0.4% Trypan blue to the fresh medium of different concentrations of drugs and corresponding solvents, count the number of living and dead cells in the four large squares with a hemocytometer, and calculate the survival rate;

It should be emphasized that when the compound is poorly soluble in water and needs to be dissolved with DMSO, the final concentration of DMSO in the MT-4 cell culture medium is less than 2%. Because DMSO can affect the anticellular activity of the test compound, the blank experiment for the anticellular activity of the solution containing the same concentration of DMSO should also be carried out in parallel. The present invention adopts the clinical drug dasatinib as a reference substance, and the results of the inhibitory activity of some target compounds on K562 cells are shown in Table 1:

Table 1. Compounds inhibit K562 cell activity

The experimental results show that the compounds contained in the general chemical structure formula generally have strong inhibitory activity on K562 cells, and compounds 4b and 6c have comparable activity to dasatinib.

Claims (3)

1.2-An Ji oxazole compounds, its structural formula is:

, wherein

R=H, C _1-5alkyl, halogen; R ₁=H, C _1-5alkyl, halogen, CN;

X=N，CH；

R ₃=H, CH ₃, halogen, CN, NO ₂;

R ₄=H, CH ₃, halogen, CN, NO ₂;

R ₂for hydroxyethyl piperazine, hydroxyethyl piperidine, piperazine ethylamino-, morpholine ethylamino-, piperidine-1-ethanamine base,

Piperazine methylamino, morpholine methylamino, piperidines methylamino, hydroxyethyl piperazine methylamino, hydroxyethyl piperidine methylamino, or C _1-5alkylamino radical.

2. the preparation method of 2-An Ji oxazole compounds described in claim 1, is characterized in that comprising the steps: substituted aromatic amines, in pyridine and tetrahydrofuran (THF), obtains E-N-aryl-3-ethoxy propylene acid amides with E-3-ethoxy propylene acyl chloride reaction; E-N-aryl-3-ethoxy propylene acid amides and NBS occur after addition reaction, close ring immediately with urea condensation, obtain formula (4 a-c) compound ; Formula (4 a-c) compound by more respectively with dichloro pyrimidine or dichloropyridine, and hydroxyethyl piperazine respectively anti-raw nucleophilic substitution obtain ultimate aim product; Or formula (4 a-c) compound is by reacting with aroyl chloride or alkyl acyl chloride, obtains ultimate aim product.

3. described in claim 1,2-An Ji oxazole compounds suppresses the application in K562 cytoactive medicine in preparation.