WO2016184122A1 - Pyrazoles compounds - Google Patents

Abstract

A new compound (as shown in formula I) and salts thereof, which are proved to have better pharmacokinetic parameters and anti-tumor activity by experiments, thereby being capable of inhibiting growth of tumor cells. Therefore, the new compound and the salts thereof are used for preparing anti-tumor drugs.

Description

[Name of invention established by ISA in accordance with Rule 37.2] A pyrazole compound Technical field

The present invention relates to an imidazole compound.

Background technique

Non-steroidal anti-inflammatory drugs (NSAIDS) show their unique anti-inflammatory, antipyretic and analgesic effects by inhibiting the synthesis of prostaglandins. Currently, the clinically widely used are aspirin, ibuprofen, ketoprofen, and flurbiol. Fenfenate, diclofenac sodium, celecoxib, parecoxib and naproxen. In the published literature, compound Z is a prodrug of celecoxib having the following structural formula:

Compound Z has better pharmacokinetic parameters than celecoxib in the published literature.

Tumors are serious diseases that endanger human health. The prevention and treatment of cancer has always been the focus of medical research. At present, due to environmental pollution caused by industrial development, the quality of human living environment is declining, and the incidence and mortality rate of cancer diseases are rising. Radiotherapy and chemotherapy are the main means of treating tumors. However, chemotherapy and radiotherapy inhibit the development of cancer cells while inhibiting the development of normal cells, reducing the body's immunity and leading to new complications. The specific drugs for treating tumor diseases are not satisfactory. Currently, the cytotoxic drugs used in clinical practice are not highly selective, resulting in malignant killing of normal cells, which limits their application. Therefore, the search for new, non-invasive, non-cytotoxic anti-tumor drugs has become an important direction in the international medical field.

Invention disclosure

Our company further synthesized compounds 1a, 1b, 1c (sodium salt of the compound of formula I) for in vitro and in vivo anti-tumor experiments and pharmacokinetic tests to compare celecoxib, compounds Z, 1a, 1b and 1c. Antitumor activity and pharmacokinetic parameters.

The series of compounds we have synthesized can be obtained by the following reaction route.

The above compounds 1a, 1b, 1c, compound Z, and celecoxib were compared for the antitumor test, and it was found that the inhibitory effect on the melanoma B16, 1c (formula I), was significantly greater than that of the other compounds. The above compounds 1a, 1b, 1c, compound Z, and celecoxib were tested for pharmacokinetics. It was found that the compounds of 1a, 1b, 1c, and Z were metabolized to different degrees in the blood after being administered by beagle dogs to celecoxib. Among them, 1c has the best pharmacokinetic characteristics, and the AUC value observed in the beagle dogs after gavage is significantly greater than other compounds.

Specific embodiment:

Example 1: Preparation of a series of compounds:

1a, 1b, 1c are obtained according to the following figure:

(1) Synthesis of Compound 3:

Accurately weigh 32.4 g of sodium methoxide and add it to 100.0 mL of DMF. Stir the reaction for 10 minutes at room temperature, cool the reaction solution to 5-10 ° C, and slowly add 67.0 g of p-methylacetophenone for 30 minutes. Continue to react for 30 minutes. 76.8 g of methyl trifluoroacetate was dissolved in 100 mL of DMF, and slowly added dropwise to the above reaction solution for 30 minutes. After the completion of the dropwise addition, the reaction was continued at room temperature for 1 hour to stop the reaction, and 300 mL of ice was added to the reaction solution. Water, pH=3-5 was adjusted with 37% hydrochloric acid, stirred well for 30 minutes, suction filtered, and the filter cake was washed with distilled water (200 mL × 3), and dried under vacuum to give 101.8 g of pale yellow compound 3 in a yield of 87.7%. H NMR (400 Hz, DMSO): 7.74 - 7.72 (d, J = 4.4 Hz, 2H), 7.18-7.16 (d, J = 4.4 Hz, 2H), 3.71 (s, 2H), 2.35 (s, 3H); (m/z): 231.3.

(2) Synthesis of Compound 4:

Accurately weigh 89.4 g of 4-(aminosulfonyl)benzoquinone and 30.0 mL of 37% hydrochloric acid into 200.0 mL of DMF, stir the reaction for 30 minutes, add 100 g of compound 3 to the reaction solution, and stir the reaction at 60 ° C for 2 hours. The reaction was stopped, and the reaction liquid was cooled to room temperature. 500 mL of distilled water was added to the reaction mixture, and the mixture was stirred for 1 hour, and suction filtered. The filter cake was washed with distilled water (200 mL×3), and dried under vacuum to obtain 149.8 g of compound 4 in a yield of 90.3. %. H NMR (400 Hz, DMSO): 7.92-7.90 (d, J = 4.8 Hz, 2H), 7.50-7.48 (d, J = 4.8 Hz, 2H), 7.37-7.35 (d, J = 4.4 Hz, 2H), 7.18 -7.16 (d, J = 4.4 Hz, 2H), 6.55 (s, 2H), 2.35 (s, 3H); MS (m/z): 382.3.

(3) Synthesis of compounds 5a, 5b, 5c:

Accurately weigh 38.1 g of compound 4 and 15.0 g of triethylamine into 200.0 mL of DCM, stir the reaction for 30 minutes, add 11.0 g of propionyl chloride to the reaction solution, add dropwise within 10 minutes, and warm to 60 ° C to stir. After reacting for 2 hours, the reaction was stopped, the reaction solution was cooled to room temperature, and 500 mL of distillation was added to the reaction liquid. The mixture was stirred for 1 hour, and the mixture was separated, and the organic layer was separated. The aqueous layer was extracted with DCM (100mL×3), and the organic phase was combined, washed with distilled water (200mL×3), and the solvent was evaporated under reduced pressure to give 35.6 g of compound. 5a, the yield was 81.3%. H NMR (400 Hz, DMSO): 8.00 (s, 1H), 7.92-7.90 (d, J = 4.8 Hz, 2H), 7.50-7.48 (d, J = 4.8 Hz, 2H), 7.37-7.35 (d, J = 4.4 Hz, 2H), 7.18-7.16 (d, J = 4.4 Hz, 2H), 6.55 (s, 2H), 2.35 (s, 3H), 2.24-2.20 (m, 2H), 1.14 (t, J = 3.6) Hz, 3H); MS (m/z): 438.3.

The above reaction procedure was repeated to obtain 36.4 g of Compound 5b in a yield of 80.3%. H NMR (400 Hz, DMSO): 8.00 (s, 1H), 7.92-7.90 (d, J = 4.8 Hz, 2H), 7.50-7.48 (d, J = 4.8 Hz, 2H), 7.37-7.35 (d, J = 4.4 Hz, 2H), 7.18-7.16 (d, J = 4.4 Hz, 2H), 6.55 (s, 2H), 2.35 (s, 3H), 2.19-2.17 (m, 2H), 1.63-1.61 (m, 2H) ), 0.96 (t, J = 3.2 Hz, 3H); MS (m/z): 453.3.

The above reaction procedure was repeated to obtain 33.8 g of Compound 5c in a yield of 74.9%. H NMR (400 Hz, DMSO): 8.00 (s, 1H), 7.92-7.90 (d, J = 4.8 Hz, 2H), 7.50-7.48 (d, J = 4.8 Hz, 2H), 7.37-7.35 (d, J = 4.4 Hz, 2H), 7.18-7.16 (d, J = 4.4 Hz, 2H), 6.55 (s, 2H), 2.35 (s, 3H), 1.19-1.17 (m, 1H), 0.71-0.68 (m, 2H) ), 0.56-0.52 (m, 2H); MS (m/z): 451.3.

(4) Synthesis of compounds 1a, 1b, 1c:

30.0 g of Compound 5a was accurately weighed and added to 100.0 mL of ethanol, and stirred at room temperature for 30 minutes. 2.9 g of sodium hydroxide was dissolved in 50.0 mL of ethanol, added dropwise to the above reaction solution, and the dropwise addition was completed within 10 minutes. The reaction was stirred at room temperature for 2 hours to stop the reaction, and 300 mL of diethyl ether was slowly added dropwise to the reaction solution. A large amount of precipitate was precipitated, stirred well for 1 hour, suction filtered, and the filter cake was washed with diethyl ether (50.0 mL × 3). After drying in vacuo, 25.3 g of Compound 1a was obtained in a yield of 80.3%. H NMR (400 Hz, DMSO): 7.92-7.90 (d, J = 4.8 Hz, 2H), 7.50-7.48 (d, J = 4.8 Hz, 2H), 7.37-7.35 (d, J = 4.4 Hz, 2H), 7.18 -7.16 (d, J = 4.4 Hz, 2H), 6.55 (s, 2H), 2.35 (s, 3H), 2.24-2.20 (m, 2H), 1.14 (t, J = 3.6 Hz, 3H); m/z): 460.4.

The above reaction procedure was repeated to obtain 24.2 g of Compound 1b in a yield of 77.1%. H NMR (400 Hz, DMSO): 7.92-7.90 (d, J = 4.8 Hz, 2H), 7.50-7.48 (d, J = 4.8 Hz, 2H), 7.37-7.35 (d, J = 4.4 Hz, 2H), 7.18 -7.16 (d, J = 4.4 Hz, 2H), 6.55 (s, 2H), 2.35 (s, 3H), 2.19-2.17 (m, 2H), 1.63-1.61 (m, 2H), 0.96 (t, J) = 3.2 Hz, 3H); MS (m/z): 474.3.

The above reaction procedure was repeated to obtain 23.7 g of Compound 1c in a yield of 75.4%. HNMR (400Hz, DMSO): 7.92-7.90 (d, J = 4.8 Hz, 2H), 7.50-7.48 (d, J = 4.8 Hz, 2H), 7.37-7.35 (d, J = 4.4 Hz, 2H), 7.18-7.16 (d , J=4.4 Hz, 2H), 6.55 (s, 2H), 2.35 (s, 3H), 1.19-1.17 (m, 1H), 0.71-0.68 (m, 2H), 0.56-0.52 (m, 2H); MS (m/z): 472.6.

Example 2: Comparison of B16 melanoma inhibition:

B16 melanoma was subcultured in subcutaneously in C57BL/6 mice. B16 melanoma-bearing mice preserved under the armpits with good tumor growth, no necrosis or liquefaction were selected, and the mice were sacrificed by cervical dislocation. After 75% alcohol soaking and disinfection, the tumor tissues were aseptically added and added 5 times volume (W/V). The physiological saline for injection was ground into a homogenate by a tissue homogenizer, and the tumor cell suspension was filtered through a sterile 200-mesh stainless steel mesh. The same as above, the C57BL/6 mice were inoculated subcutaneously and routinely reared.

Grouping and administration: 70 tumor-bearing mice were randomly divided into 5 groups according to their body weight, 10 in each group, which were model group, cyclophosphamide group, 1a group, 1b group, 1c group, compound Z group, and Sailai. In the Xibu group, the dose was 60 mg/kg, and the solution was prepared into a 3 mg/ml solution using physiological saline. Each group of mice was administered at the dose and mode shown in Table 3. The cyclophosphamide group was given intraperitoneal administration of cyclophosphamide only once on the second day of tumor-bearing, and each group in the experimental group was administered once per day for 10 consecutive times. day. The administration volume was 20 ml/kg body weight. 24 hours after the last administration, the mice were sacrificed by cervical dislocation, the body weight was weighed, the tumor tissue was removed, and the tumor inhibition rate was calculated.

表示，以t检验进行组间统计学差异比较。Result

The t test was used to compare the statistical differences between the groups.

The results showed that the five experimental groups received intravenous injection for 10 days, which all inhibited the growth of transplanted tumor B16 melanoma in mice, and the compound 1c group was optimal.

Table 1: Inhibition of mouse B16 melanoma growth

Compared with the model group: ^* , P <0.05; ^** , P < 0.01.

Example 3: Pharmacokinetic test

Ten male beagle dogs, 2 in each group, were fasted to water one night before the experiment and weighed before administration.

(A) celecoxib group

(B) Compound Z group

(C) Compound 1a group

(D) Compound 1b group

(E) Compound 1c group

Dosing: 200 mg of celecoxib was used, and the other groups were calculated to be equivalent to the mass of 200 mg of celecoxib, and 0.5 ml of CMC was used to prepare a 10 ml suspension. After a single administration of 10 ml of the solution, 10 ml of water was supplied. Detection conditions: using C18 column, 150mm × 4.6mm, 5um, mobile phase is 30mmol / L ammonium acetate-methanol (22:78V / V), flow rate: 1ml / min, column temperature: 40 ° C, detection wavelength: 254nm . Blood samples were taken at 0-24 hours after administration, and blood plasma concentrations of celecoxib were measured by validated high performance liquid chromatography. Table 2: Plasma drug metabolism of each group in celecoxib The learning parameters are as follows:

Claims (3)

a compound of the structure shown in formula (I):

A sodium salt of a compound according to claim 1. Use of a compound according to claims 1 and 2 for the manufacture of a medicament for the treatment of tumors.