Dianilinopyrimidine compound for inhibiting kinase activity

Abstract

Disclosed are a dianilino pyrimidine compound having an inhibitory effect on protein tyrosine kinase, pharmaceutically acceptable salts, crystal forms, prodrugs, metabolites, hydrates, solvates, stereoisomers or isotopic derivatives thereof, a pharmaceutical composition containing these compounds, as well as preparation and use of these compounds. The compound has a structure as represented by formula (I), and may be used for treating ALK-mediated cancer-related symptoms, such as non-small cell lung cancer, breast cancer, nerve tumors, esophagus cancer, soft tissue cancer, lymphoma, or leukemia. ##STR00001##

Claims

1. A compound of formula (I): ##STR00072## wherein, R.sub.1 and R.sub.2 are independently selected from H, halogen, —CN, —NO.sub.2, —OH, optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.1-C.sub.6 haloalkyl, optionally substituted C.sub.1-C.sub.6 alkoxy, optionally substituted C.sub.3-C.sub.10 carbocyclyl, and optionally substituted 3- to 10-membered heterocyclyl; linker W is C.sub.1-C.sub.6 alkylene which is optionally substituted with one or more R.sub.3; R.sub.3 is selected from H, halogen, —CN, C.sub.1-C.sub.6 alkyl, and C.sub.1-C.sub.6 haloalkyl; R.sub.5 and R.sub.6 are independently selected from H, optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.1-C.sub.6 haloalkyl, optionally substituted C.sub.1-C.sub.6 alkoxy, optionally substituted C.sub.3-C.sub.10 carbocyclyl and optionally substituted 3- to 10-membered heterocyclyl; R.sub.7 is selected from optionally substituted C.sub.1-C.sub.6 alkoxy, optionally substituted C.sub.1-C.sub.6 haloalkoxy, optionally substituted C.sub.2-C.sub.6 alkenyloxy, and optionally substituted 3- to 7-membered cycloalkyloxy; and R.sub.8 is selected from H, halogen, and C.sub.1-C.sub.6 alkyl; or a pharmaceutically acceptable salt, a crystal form, a prodrug, a metabolite, a hydrate, a solvate, a stereoisomer, or an isotopic derivative thereof.

2. The compound according to claim 1, wherein R.sub.1 is selected from H, halogen, —CN, optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.1-C.sub.6 haloalkyl, optionally substituted C.sub.1-C.sub.6 alkoxy, optionally substituted C.sub.3-C.sub.10 carbocyclyl, and optionally substituted 3- to 10-membered heterocyclyl; and R.sub.2 is selected from H, halogen, —CN, —NO.sub.2, —OH, optionally substituted C.sub.1-C.sub.6 alkyl, and optionally substituted C.sub.1-C.sub.6 alkoxy; or a pharmaceutically acceptable salt, a crystal form, a prodrug, a metabolite, a hydrate, a solvate, a stereoisomer, or an isotopic derivative thereof.

3. The compound according to claim 2, which is of the following formula: ##STR00073## wherein, W is C.sub.1-C.sub.6 alkylene which is optionally substituted with one or more R.sub.3; R.sub.3 is selected from H, C.sub.1-C.sub.6 alkyl, and C.sub.1-C.sub.6 haloalkyl; R.sub.5 and R.sub.6 are independently selected from H, optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.1-C.sub.6 haloalkyl, optionally substituted C.sub.1-C.sub.6 alkoxy, optionally substituted C.sub.3-C.sub.10 carbocyclyl, and optionally substituted 3- to 10-membered heterocyclyl; R.sub.7 is selected from optionally substituted C.sub.1-C.sub.6 alkoxy, optionally substituted C.sub.1-C.sub.6 haloalkoxy, and optionally substituted 3- to 7-membered cycloalkyloxy; and R.sub.8 is selected from H, halogen, and C.sub.1-C.sub.6 alkyl; or a pharmaceutically acceptable salt, a crystal form, a prodrug, a metabolite, a hydrate, a solvate, a stereoisomer, or an isotopic derivative thereof.

4. The compound according to claim 1, wherein W is selected from —CH.sub.2—, —CH.sub.2CH.sub.2—, —C(CH.sub.3)H—, —C(CH.sub.3).sub.2—, —C(CH.sub.3)HCH.sub.2—, —C(CH.sub.3).sub.2CH.sub.2—, —C(CH.sub.3)HC(CH.sub.3)H—, and —C(CH.sub.3).sub.2C(CH.sub.3).sub.2; or a pharmaceutically acceptable salt, a crystal form, a prodrug, a metabolite, a hydrate, a solvate, a stereoisomer, or an isotopic derivative thereof.

5. The compound according to claim 1, wherein R.sub.5 and R.sub.6 are independently selected from H, and optionally substituted C.sub.1-C.sub.6 alkyl; or a pharmaceutically acceptable salt, a crystal form, a prodrug, a metabolite, a hydrate, a solvate, a stereoisomer, or an isotopic derivative thereof.

6. The compound according to claim 1, wherein —W—NR.sub.5R.sub.6 is selected from: ##STR00074## or a pharmaceutically acceptable salt, a crystal form, a prodrug, a metabolite, a hydrate, a solvate, a stereoisomer, or an isotopic derivative thereof.

7. The compound according to claim 1, wherein R.sub.7 is selected from —OCH.sub.3, —OCH.sub.2CH.sub.3, —OCH.sub.2CH.sub.2CH.sub.3, —OCH(CH.sub.3).sub.2, —OCF.sub.3, —OCHF.sub.2, —OCH.sub.2F, and —OCH.sub.2CF.sub.3; or a pharmaceutically acceptable salt, a crystal form, a prodrug, a metabolite, a hydrate, a solvate, a stereoisomer, or an isotopic derivative thereof.

8. The compound according to claim 1, wherein R.sub.8 is selected from H and C.sub.1-C.sub.6 alkyl; or a pharmaceutically acceptable salt, a crystal form, a prodrug, a metabolite, a hydrate, a solvate, a stereoisomer, or an isotopic derivative thereof.

9. The compound according to claim 1, wherein the compound is selected from: ##STR00075## ##STR00076## ##STR00077## ##STR00078## ##STR00079## or a pharmaceutically acceptable salt, a crystal form, a prodrug, a metabolite, a hydrate, a solvate, a stereoisomer or an isotopic derivative thereof.

10. A pharmaceutical composition containing the compound according to claim 1, or a pharmaceutically acceptable salt, a crystal form, a prodrug, a metabolite, a hydrate, a solvate, a stereoisomer, or an isotopic derivative thereof, and pharmaceutically acceptable excipient(s).

11. The compound according to claim 1, wherein R.sub.1 is Cl; or a pharmaceutically acceptable salt, a crystal form, a prodrug, a metabolite, a hydrate, a solvate, a stereoisomer, or an isotopic derivative thereof.

12. The compound according to claim 1, wherein R.sub.2 is H; or a pharmaceutically acceptable salt, a crystal form, a prodrug, a metabolite, a hydrate, a solvate, a stereoisomer, or an isotopic derivative thereof.

13. The compound according to claim 1, wherein R.sub.7 is —OCH.sub.3; or a pharmaceutically acceptable salt, a crystal form, a prodrug, a metabolite, a hydrate, a solvate, a stereoisomer, or an isotopic derivative thereof.

14. The compound according to claim 1, wherein R.sub.8 is selected from H and methyl; or a pharmaceutically acceptable salt, a crystal form, a prodrug, a metabolite, a hydrate, a solvate, a stereoisomer, or an isotopic derivative thereof.

15. A compound, wherein the compound is selected from: ##STR00080## or a pharmaceutically acceptable salt, a crystal form, a prodrug, a metabolite, a hydrate, a solvate, a stereoisomer, or an isotopic derivative thereof.

Description

EXAMPLES

(1) The present disclosure will be further described below in combination with specific embodiments. It should be understood that these examples are only used to illustrate the present disclosure and not to limit the scope of the present disclosure. The experimental methods without specific conditions in the following examples generally follow the conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, parts and percentages are parts by weight and weight percent.

(2) Generally, in the preparation process, each reaction is usually carried out in an inert solvent at a temperature from room temperature to reflux temperature (such as 0° C. to 100° C., alternatively 0° C. to 80° C.). The reaction time is usually 0.1-60 hours, alternatively 0.5-24 hours.

Example 1 Preparation of (2-((5-chloro-2-((4-((dimethylamino)methyl)-2-methoxyphenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide (Compound T-1)

(3) ##STR00034##

(4) The following route was used for the synthesis

(5) ##STR00035## ##STR00036##

(6) Step 1 Synthesis of Compound 2

(7) Compound 1 (0.6 g, 3.0 mmol) and methanol (15 mL) were added in sequence into a 50 mL single-necked flask equipped with a magnetic stirrer, and the mixture was stirred to form a solution. Concentrated sulfuric acid (1 mL) was slowly added dropwise. After the addition was completed, the temperature was raised to reflux temperature and the reaction was stirred at this temperature under a nitrogen atmosphere for 3 hours. After cooled to room temperature, the reaction was quench by adding water (40 mL), and the resulting solution was extracted with dichloromethane (40 mL×2). Organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give 0.52 g of colorless liquid. Yield: 82.1%. LC-MS(APCI): m/z=212.1 (M+1).sup.+.

(8) Step 2 Synthesis of Compound 3

(9) To a 50 mL single-necked flask equipped with a magnetic stirrer compound 2 (0.5 g, 2.37 mmol) and anhydrous THF (15 mL) were added in sequence, and the mixture was stirred to form a solution. After cooled in an ice water bath, lithium aluminum hydride (LiAlH.sub.4, 90 mg, 2.37 mmol) was slowly added, and the reaction was stirred at this temperature under a nitrogen atmosphere for 1 hour. Sodium sulfate decahydrate (10 g) was added to quench the reaction, which was diluted with dichloromethane (40 mL) and filtered after stirring for 5 minutes. The filter cake was washed with dichloromethane (10 mL), the resulting solution was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 0.4 g of a white solid. Yield: 92.2%. LC-MS(APCI): m/z=184.1 (M+1).sup.+.

(10) Step 3 Synthesis of Compound 4

(11) To a 100 mL single-necked flask equipped with a magnetic stirrer compound 3 (400 mg, 2.18 mmol) and dichloromethane (DCM, 10 mL) were added in sequence, and the mixture was stirred to form a solution. Dess-Martin periodinane (904 mg, 2.18 mmol) was added and the reaction mixture was stirred at room temperature for 1 hour under a nitrogen atmosphere. The reaction was quenched by adding water (20 mL), the resulting solid was filtered, and the aqueous phase was extracted with dichloromethane (20 mL×2). The organic phases were combined, washed with water (30 mL×2), dried over anhydrous sodium sulfate, concentrated and passed through a silica gel column to obtain 300 mg of a white solid. Yield: 74.9%. LC-MS(APCI): m/z=182.1 (M+1).sup.+.

(12) Step 4 Synthesis of Compound 5

(13) To a 50 mL single-necked flask equipped with a magnetic stirrer compound 4 (181 mg, 1.0 mmol) and methanol (5 ml) were added in sequence, and the mixture was stirred to form a solution. A solution of dimethylamine in tetrahydrofuran (1 mL, 2.0 mmol, 2M) and glacial acetic acid (1 drop) were added dropwise, and the reaction was stirred at room temperature for 10 minutes under a nitrogen atmosphere, and NaBH.sub.3CN (150 mg, 2.5 mmol) was slowly added in dropwise, and the reaction was stirred at room temperature for 1 hour. The reaction was quenched by adding water (5 mL), and extracted with ethyl acetate (15 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was passed through a silica gel column to obtain 126 mg of a white solid. Yield: 60.0%. LC-MS(APCI): m/z=211.1 (M+1).sup.+.

(14) Step 5 Synthesis of Compound 6

(15) To a 50 mL single-necked flask equipped with a magnetic stirrer and a condenser tube, was added compound 5 (126 mg, 0.6 mmol) and ethanol/water (8 mL, 3/1), and the mixture was stirred to form a solution. Reduced iron powder (3.6 mmol, 202 mg) and ammonium chloride (0.6 mmol, 32 mg) was added, the reaction was heated to reflux under a nitrogen atmosphere and reacted for 1 hour. Then the reaction was cooled to room temperature, filtered and the filter cake was washed with ethanol (5 mL). The organic solvent was removed by concentration, and the resulting solution was extracted with dichloromethane (10 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give 80 mg of a brown solid. Yield: 74.1%. LC-MS(APCI): m/z=181.1 (M+1).sup.+.

(16) Step 6 Synthesis of compound T-1

(17) To a 25 mL single-necked flask equipped with a magnetic stirrer and a condenser tube, compound 6 (80 mg, 0.44 mmol), compound 7 (138 mg, 0.44 mmol) and ethylene glycol monomethyl ether (3 mL) were added, and the mixture was stirred to form a solution.

(18) A solution of hydrogen chloride in isopropanol (0.66 mmol, 0.13 mL, 5M) was added dropwise, and the temperature was raised to 120° C. under a nitrogen atmosphere and the reaction was stirred at this temperature overnight. The reaction was cooled to room temperature, water (10 mL) and saturated sodium bicarbonate (5 mL) was added, the solution was extracted with dichloromethane (15 mL×3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was passed through a silica gel column to give 100 mg of a white solid. Yield: 49.5%. LC-MS (APCI): m/z=460.2 (M+1).sup.+. .sup.1H NMR (400 MHz, DMSO-D.sub.6) δ ppm: 11.17 (s, 1H), 8.47-8.43 (m, 1H), 8.14 (s, 1H), 8.12 (s, 1H), 7.71 (d, J=8.0 Hz, 1H), 7.58-7.53 (m, 1H), 7.39 (t, J=8.0 Hz, 1H), 7.13 (t, J=8.0 Hz, 1H), 6.97 (s, 1H), 6.81 (d, J=8.0 Hz, 1H), 3.80 (s, 3H), 2.17 (s, 6H), 1.78 (s, 3H), 1.75 (s, 3H).

Example 2 Preparation of (2-((2-((4-(2-aminoethyl)-2-methoxyphenyl)amino)-5-chloropyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide (Compound T-2)

(19) ##STR00037##

(20) The following route was used for the synthesis:

(21) ##STR00038## ##STR00039##

(22) Step 1 Synthesis of Compound 10

(23) To a 250 mL single-necked flask equipped with a magnetic stirrer compound 8 (5.1 g, 30 mmol) and anhydrous DMSO (40 mL) were added in sequence, the mixture was stirred to form a solution under a nitrogen atmosphere, and after cooled to 0° C., NaH (3.6 g, 90 mmol, 60%) was slowly added, and the solution was stirred at 0° C. for half an hour. A solution of compound 9 (11.6 g, 90 mmol) in DMSO (10 mL) was added dropwise, the ice bath was removed, and the reaction mixture was warmed to 100° C. and the reaction was stirred at this temperature for 3 hours. The reaction was cooled to room temperature before the addition of water (50 mL), and the solution was extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with water (60 mL×2), dried over anhydrous sodium sulfate, filtered, concentrated, and passed through a silica gel column to give 8.3 g of a white solid.

(24) Yield: 98.1%. LC-MS(APCI): m/z=282.1 (M−1).sup.−.

(25) Step 2 Synthesis of Compound 11

(26) To a 250 mL single-necked flask equipped with a magnetic stirrer was added compound 10 (2.83 g, 10 mmol) and DMSO (50 mL) in sequence, and the mixture was stirred to form a solution. NaCl (1.17 g, 20 mmol) and water (0.18 g, 10 mmol) were added. The reaction mixture was heated to 120° C. and stirred at this temperature for 3 hours. The reaction was cooled to room temperature, added with water (80 mL), extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with water (60 mL×2), dried over anhydrous sodium sulfate, filtered, concentrated and passed through a silica gel column to obtain 1.9 g of a white solid. Yield: 84.1%. LC-MS(APCI): m/z=224.1 (M−1).sup.−.

(27) Step 3 Synthesis of compound 12

(28) To a 250 mL single-necked flask equipped with a magnetic stirrer compound 11 (1.9 g, 8.4 mmol) and anhydrous THF (50 mL) were added in sequence, and the mixture was stirred to form a solution. The solution was cooled in an ice water bath, and LiAlH.sub.4 (320 mg, 8.4 mmol) was slowly added. The reaction was stirred at this temperature under a nitrogen atmosphere for 1 hour. The reaction was quenched by adding sodium sulfate decahydrate (20 g), and stirred for 5 minutes. The solution was diluted with dichloromethane (60 mL), filtered, and the filter cake was washed with dichloromethane (10 mL). The resulting solution was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 1.3 g of a white solid. Yield: 79.0%. LC-MS(APCI): m/z=198.1 (M+1).sup.+.

(29) Step 4 Synthesis of compound 13

(30) To a 100 mL single-necked flask equipped with a magnetic stirrer and a condenser tube was added compound 12 (1.3 g, 6.6 mmol) and ethanol/water (40 mL, 3/1), and the mixture was stirred to form a solution. Reduced iron powder (39.6 mmol, 2.23 g) and ammonium chloride (6.6 mmol, 350 mg) were added, the reaction was heated to reflux under a nitrogen atmosphere and reacted at this temperature for 1 hour. The reaction was cooled to room temperature, filtered, the filter cake was washed with ethanol (10 mL). The organic solvent was removed by concentration, and the resulting solution was extracted with dichloromethane (30 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give 850 mg of a brown solid. Yield: 77.1%. LC-MS(APCI): m/z=168.1 (M+1).sup.+.

(31) Step 6 Synthesis of compound 14

(32) To a 50 mL single-necked flask equipped with a magnetic stirrer and a condenser tube compound 13 (850 mg, 5.09 mmol), compound 7 (1.6 g, 5.09 mmol) and ethylene glycol monomethyl ether (10 mL) were added, and the mixture was stirred to form a solution. A solution of hydrogen chloride in isopropanol (7.64 mmol, 1.53 mL, 5M) was added dropwise, and the temperature was raised to 120° C. under a nitrogen atmosphere, and the reaction was stirred at this temperature overnight. The reaction was cooled to room temperature, water (30 mL) and saturated sodium bicarbonate (20 mL) were added, and the solution was extracted with dichloromethane (50 mL×3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, the residue was passed through a silica gel column to give 1.2 g of a white solid. Yield: of 52.9%. LC-MS(APCI): m/z=447.2 (M+1).sup.+. .sup.1H NMR (400 MHz, DMSO-D.sub.6) 5/ppm: 11.16 (s, 1H), 8.46-8.43 (m, 1H), 8.1 l(s, 1H), 8.07 (s, 1H), 7.62 (d, J=8.0 Hz, 1H), 7.58-7.51 (m, 1H), 7.39 (t, J=8.0 Hz, 1H), 7.12 (t, J=8.0 Hz, 1H), 6.90 (s, 1H), 6.73 (d, J=8.0 Hz, 1H), 4.62 (t, J=4.8 Hz, 1H), 3.77 (s, 3H), 3.61 (q, J=4.8 Hz, 2H), 2.71 (t, J=5.6 Hz, 2H), 1.77 (s, 3H), 1.74 (s, 3H).

(33) Step 7 Synthesis of compound 15

(34) To a 100 mL single-necked flask equipped with a magnetic stirrer and a condenser tube compound 14 (1.7 g, 3.8 mmol) and anhydrous dichloromethane (20 mL) were added, and the mixture was stirred to form a solution. CBr.sub.4 (4.1 mmol, 1.36 g) was added, the solution was cooled to 0° C. under a nitrogen atmosphere, and triphenylphosphine (TPP, 1.08 g, 4.1 mmol) was added. The ice bath was removed after addition, and the reaction was stirred at room temperature for 1 h. The solvent was distilled off under reduced pressure, and the residue was passed through a silica gel column to obtain 960 mg of a white solid. Yield: 49.5%. LC-MS(APCI): m/z=509.1, 511.1 (M+1).sup.+.

(35) Step 8 Synthesis of Compound T-2

(36) To a 50 mL single-necked flask equipped with a magnetic stirrer Compound 15 (102 mg, 0.2 mmol) and acetonitrile (2 mL) were added, and the mixture was stirred to form a solution. Ammonia (3 mL) was added, and the reaction was stirred at room temperature under a nitrogen atmosphere overnight. The solvent was distilled off under reduced pressure, and the residue was passed through a silica gel column to obtain 60 mg of a white solid. Yield: 67.4%. LC-MS(APCI): m/z=446.1 (M+1).sup.+. .sup.1H NMR (400 MHz, DMSO-D.sub.6) δ ppm: 11.19 (s, 1H), 8.49-8.46 (m, 1H), 8.15 (s, 1H), 8.10 (s, 1H), 7.96 (br s, 2H), 7.77 (d, J=8.0 Hz, 1H), 7.61-7.55 (m, 1H), 7.45 (t, J=8.0 Hz, 1H), 7.16 (t, J=8.0 Hz, 1H), 6.97 (s, 1H), 6.78 (d, J=8.0 Hz, 1H), 3.84 (s, 3H), 3.06 (t, J=8.0 Hz, 2H), 2.88 (t, J=8.0 Hz, 2H), 1.80 (s, 3H), 1.76 (s, 3H).

Example 3 Preparation of (2-((5-chloro-2-((4-(2-(dimethylamino)ethyl)-2-methoxyphenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide (Compound T-3)

(37) ##STR00040##

(38) The following route was used for the synthesis:

(39) ##STR00041##

(40) To a 50 mL single-necked flask equipped with a magnetic stirrer compound 15 (102 mg, 0.2 mmol) and acetonitrile (2 mL) were added, and the mixture was stirred to form a solution. A solution of dimethylamine in tetrahydrofuran (5 mL) was added, and the reaction was stirred at room temperature under a nitrogen atmosphere overnight. The solvent was distilled off under reduced pressure, and the residue was passed through a silica gel column to obtain 60 mg of a white solid. Yield: 63.4%. LC-MS(APCI): m/z=474.2 (M+1).sup.+. .sup.1H NMR (400 MHz, DMSO-D.sub.6) δ ppm: 11.19 (s, 1H), 8.48-8.45 (m, 1H), 8.14 (s, 1H), 8.12 (s, 1H), 7.71 (d, J=8.0 Hz, 1H), 7.60-7.56 (m, 1H), 7.44 (t, J=8.0 Hz, 1H), 7.15 (t, J=8.0 Hz, 1H), 6.99 (s, 1H), 6.79 (d, J=8.0 Hz, 1H), 3.82 (s, 3H), 3.04-3.00 (m, 2H), 2.95-2.89 (m, 2H), 2.60 (s, 6H), 1.80 (s, 3H), 1.76 (s, 3H).

Example 4 Preparation of (2-((5-chloro-2-((2-methoxy-4-(2-(piperidin-1-yl)ethyl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide (Compound T-4)

(41) ##STR00042##

(42) The following route was used for the synthesis:

(43) ##STR00043##

(44) To a 50 mL single-necked flask equipped with a magnetic stirrer compound 15 (102 mg, 0.2 mmol) and acetonitrile (2 mL) were added, and the mixture was stirred to form a solution. Piperidine (51 mg, 0.6 mmol) was added, and the reaction was stirred at room temperature under a nitrogen atmosphere overnight. The solvent was distilled off under reduced pressure, and the residue was passed through a silica gel column to obtain 60 mg of a white solid. Yield: 58.5%. LC-MS(APCI): m/z=514.2 (M+1).sup.+. .sup.1H NMR (400 MHz, DMSO-D.sub.6) δ ppm: 11.19 (s, 1H), 8.48-8.45 (m, 1H), 8.14 (s, 1H), 8.12 (s, 1H), 7.69 (d, J=8.0 Hz, 1H), 7.60-7.56 (m, 1H), 7.43 (t, J=8.0 Hz, 1H), 7.15 (t, J=8.0 Hz, 1H), 6.97 (s, 1H), 6.78 (d, J=8.0 Hz, 1H), 3.81 (s, 3H), 3.38-3.36 (m, 2H), 2.91-2.62 (m, 6H), 1.80 (s, 3H), 1.76 (s, 3H), 1.69-1.62 (m, 4H), 1.49-1.45 (m, 2H).

Example 5 Preparation of (2-((5-chloro-2-((2-methoxy-4-(2-(4-methylpiperazin-1-yl)ethyl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide (Compound T-5)

(45) ##STR00044##

(46) The following route was used for the synthesis:

(47) ##STR00045##

(48) To a 50 mL single-necked flask equipped with a magnetic stirrer compound 15 (102 mg, 0.2 mmol) and acetonitrile (2 mL) were added, and the mixture was stirred to form a solution. N-methylpiperazine (60 mg, 0.6 mmol) was added, the reaction was stirred at room temperature under a nitrogen atmosphere overnight. The solvent was distilled off under reduced pressure, and the residue was passed through a silica gel column to obtain 60 mg of a white solid. Yield: 56.8%. LC-MS(APCI): m/z=514.2 (M+1).sup.+. .sup.1H NMR (400 MHz, DMSO-D.sub.6) δ ppm: 11.18 (s, 1H), 8.48-8.45 (m, 1H), 8.13 (s, 1H), 8.10 (s, 1H), 7.64 (d, J=8.0 Hz, 1H), 7.59-7.54 (m, 1H), 7.41 (t, J=8.0 Hz, 1H), 7.14 (t, J=8.0 Hz, 1H), 6.96 (s, 1H), 6.76 (d, J=8.0 Hz, 1H), 3.80 (s, 3H), 3.38-3.30 (m, 4H), 2.74 (t, J=8.0 Hz, 2H), 2.59-2.52 (m, 4H), 2.50-2.47 (m, 2H), 2.26 (s, 3H), 1.79 (s, 3H), 1.76 (s, 3H).

Example 6 Preparation of (2-((5-chloro-2-((2-methoxy-4-(2-morpholinoethyl)phenyl)amino)pyrimidin-4-yl)amino) phenyl)dimethylphosphine oxide (Compound T-6)

(49) ##STR00046##

(50) The following route was used for the synthesis:

(51) ##STR00047##

(52) To a 50 mL single-necked flask equipped with a magnetic stirrer compound 15 (102 mg, 0.2 mmol) and acetonitrile (2 mL) were added, and the mixture was stirred to form a solution, morpholine (52 mg, 0.6 mmol) was added, and the reaction was stirred at room temperature under a nitrogen atmosphere overnight. The solvent was distilled off under reduced pressure, and the residue was passed through a silica gel column to obtain 60 mg of a white solid. Yield: 58.2%. LC-MS(APCI): m/z=516.2 (M+1).sup.+. .sup.1H NMR (400 MHz, DMSO-D.sub.6) δ ppm: 11.18 (s, 1H), 8.48-8.45 (m, 1H), 8.13 (s, 1H), 8.10 (s, 1H), 7.64 (d, J=8.0 Hz, 1H), 7.59-7.54 (m, 1H), 7.41 (t, J=8.0 Hz, 1H), 7.14 (t, J=8.0 Hz, 1H), 6.96 (s, 1H), 6.76 (d, J=8.0 Hz, 1H), 3.80 (s, 3H), 3.60 (t, J=8.4 Hz, 4H), 2.74 (t, J=8.0 Hz, 2H), 2.54 (t, J=8.0 Hz, 2H), 2.45 (t, J=8.0 Hz, 4H), 1.79 (s, 3H), 1.76 (s, 3H).

Example 7 Preparation of (2-((2-((4-(1-aminoeth yl)-2-methoxyphen yl)amino)-5-chloropyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide (Compound T-7)

(53) ##STR00048##

(54) The following route was used for the synthesis:

(55) ##STR00049## ##STR00050##

(56) Step 1 Synthesis of compound 17

(57) To a 100 mL three-neck flask equipped with a magnetic stirrer compound 1 (2.75 g, 13.95 mmol) and anhydrous THF (20 mL) were added in sequence, and the mixture was stirred to form a solution. After cooled to 0° C. under a nitrogen atmosphere, anhydrous DMF was added dropwise (3 drops), and then, a solution of oxalyl chloride in dichloromethane (16.74 mmol, 8.37 mL, 2M) was slowly added dropwise. After the addition, the ice bath was removed, and the reaction was stirred at room temperature for 1 hour. The solvent was distilled off under reduced pressure and the residue was dissolved in toluene (40 mL). Under stirring and under nitrogen, MgCl.sub.2 (927 mg, 9.74 mmol) and triethylamine (3.38 g, 334.4 mmol) were added with stirring under a nitrogen atmosphere, and then compound 9 (2.21 g, 16.7 mmol) was slowly added, the reaction mixture was stirred at room temperature overnight. The solvent was distilled off under reduced pressure, and the residue was passed through a silica gel column to obtain 1.68 g of a light yellow solid. Yield: 61.7%. LC-MS(APCI): m/z=196.1 (M+1).sup.+.

(58) Step 2 Synthesis of Compound 18

(59) To a 20 mL microwave tube equipped with a magnetic stirrer compound 17 (500 mg, 2.56 mmol) and ethanol (10 mL) were added, and the mixture was stirred to form a solution. Ammonium acetate (1.97 g, 25.62 mmol) and NaBFLCN (322 mg, 5.12 mmol) were added, the reaction mixture was placed in a microwave reactor and heated to a temperature of 120° C., and the reaction was stirred for 10 minutes. The reaction was cooled to room temperature, the solvent was distilled off, and the residue was passed through a silica gel column to obtain 500 mg of colorless oil. Yield: 99.5%. LC-MS(APCI): m/z=197.1 (M+1).sup.+.

(60) Step 3 Synthesis of Compound 19

(61) To a 50 mL single-necked flask equipped with a magnetic stirrer compound 18 (500 mg, 2.55 mmol) and dichloromethane (10 mL) were added, and the mixture was stirred to form a solution. Triethylamine (1.29 g, 12.74 mmol) was added, and trifluoroacetic anhydride (TFAA, 1.07 g, 5.1 mmol) was added dropwise to the solution under ice-water bath, after the addition was finished, the ice bath was removed, and the reaction was stirred at room temperature for 30 minutes under a nitrogen atmosphere. The solvent was distilled off under reduced pressure, and the residue was passed through a silica gel column to obtain 230 mg of a pale yellow solid. Yield: 30.9%. LC-MS(APCI): m/z=293.2 (M+1).sup.+. .sup.1H NMR (300 MHz, CDCl.sub.3) δ (ppm): 7.88 (d, J=6.3 Hz, 1H), 7.04 (d, J=1.2 Hz, 1H), 6.99 (dd, J=6.6 Hz, J=1.2 Hz, 1H), 5.16 (t, J=5.4 Hz, 1H), 3.99 (s, 3H), 1.63 (t, J=5.4 Hz, 3H).

(62) Step 4 Synthesis of Compound 20

(63) To a 50 mL single-necked flask equipped with a magnetic stirrer compound 19 (230 mg, 0.79 mmol) and methanol (10 mL) were added, and the mixture was stirred to form a solution. Pd/C (50 mg, 60%) was added, then the reaction was vacuumed and purged with hydrogen three times. The reaction was stirred overnight at room temperature under a balloon of hydrogen gas. The catalyst was filtered off, the filter cake was washed with methanol (5 mL), and the solvent was distilled off to obtain 180 mg of a yellow solid. Yield: 87.2% LC-MS(APCI): m/z=263.2 (M+1).sup.+.

(64) Step 5 Synthesis of Compound 21

(65) To a 50 mL single-necked flask equipped with a magnetic stirrer and a condenser tube compound 20 (180 mg, 0.69 mmol), compound 7 (239 mg, 0.75 mmol) and ethylene glycol monomethyl ether (3 mL) were added, and the mixture was stirred to form a solution. A solution of hydrogen chloride in isopropanol (1.03 mmol, 0.21 mL, 5M) was added dropwise, and the temperature was raised to 120° C. under a nitrogen atmosphere and the reaction was stirred at this temperature overnight. The reaction was cooled to room temperature, water (10 mL) and saturated sodium bicarbonate (5 mL) were added, and the solution was extracted with dichloromethane (20 mL×3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was passed through a silica gel column to give 0.3 g of a white solid. Yield: 80.6%. LC-MS(APCI): m/z=542.2 (M+1).sup.+.

(66) Step 6 Synthesis of Compound T-7

(67) To a 50 mL single-necked flask equipped with a magnetic stirrer compound 21 (300 mg, 0.55 mmol) and THF/MeOH/H.sub.2O (2 mL/1 mL/1 mL) were added, and the mixture was stirred to form a solution. Lithium hydroxide monohydrate (LiOH.H.sub.2O, 116 mg, 2.77 mmol) was added, and the reaction mixture was stirred overnight at room temperature. Water (10 mL) was added, and the solution was extracted with dichloromethane (15 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was passed through a silica gel column to obtain 0.2 g of a white solid. Yield: 81.0%. LC-MS(APCI): m/z=446.3 (M+1).sup.+. .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ (ppm): 11.19 (s, 1H), 8.47 (dd, J=6.3 Hz, J=3.3 Hz, 1H), 8.15 (s, 2H), 8.03 (br s, 2H), 7.84 (d, J=6.3 Hz, 1H), 7.56 (dd, J=10.2 Hz, J=5.7 Hz, 1H), 7.48 (t, J=5.7 Hz, 1H), 7.26 (s, 1H), 7.15 (t, J=5.4 Hz, 1H), 6.399 (d, J=6.0 Hz, 1H), 4.33 (d, J=5.1 Hz, 1H), 3.84 (s, 3H), 1.76 (d, J=9.9 Hz, 6H), 1.50 (d, J=5.1 Hz, 3H).

Example 8 and Example 9 Preparation of (2-((5-chloro-2-((4-(1-(dimethylamino)ethyl)-2-methoxyphenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide (Compound T-8) and (2-((5-chloro-2-((4-(1-(methylamino)ethyl)-2-methoxyphenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide (Compound T-9)

(68) ##STR00051##

(69) The following route was used for the synthesis:

(70) ##STR00052##

(71) To a 50 mL single-necked flask equipped with a magnetic stirrer compound T-7 (110 mg, 0.25 mmol) and methanol (5 mL), and the mixture was stirred to form a solution. Formaldehyde (14 mg, 0.17 mmol) and glacial acetic acid (1 drop) were added. The reaction was stirred for 10 minutes under a nitrogen atmosphere. The reaction was cooled to 0° C. before NaBH.sub.3CN (15 mg, 0.25 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. The reaction was quenched by water (10 mL), and extracted with dichloromethane (15 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was passed through a silica gel column to obtain T-8, as 30 mg of a white solid. Yield: 25.7%. LC-MS(APCI): m/z=474.2 (M+1).sup.+. .sup.1H NMR (300 MHz, CDCl.sub.3) δ (ppm): 10.87 (s, 1H), 8.58 (dd, J=6.6 Hz, J=3.3 Hz, 1H), 8.41 (d, J=6.3 Hz, 1H), 8.15 (s, 1H), 7.61-7.55 (m, 2H), 7.34-7.31 (m, 2H), 7.18 (t, J=5.4 Hz, 1H), 6.87 (d, J=6.3 Hz, 1H), 4.00 (s, 3H), 3.87 (s, 1H), 2.59 (s, 6H), 1.85 (s, J=9.9 Hz, 6H), 1.68 (s, 3H);

(72) The residue was passed through a silica gel column to obtain T-9 as 17 mg of a white solid. Yield: 15%. LC-MS(APCI): m/z=460.2 (M+1).sup.+. .sup.1H NMR (300 MHz, CDCl.sub.3) δ (ppm): 10.84 (s, 1H), 8.56 (dd, J=6.3z, J=3.0z, 1H), 8.42 (d, J=6.3 Hz, 1H), 8.14 (s, 1H), 7.62-7.58 (m, 2H), 7.36-7.27 (m, 2H), 7.19 (dd, J1=5.4 Hz, J2=0.9 Hz, 1H), 6.91 (d, J=6.3 Hz, 1H), 4.08 (d, J=5.1 Hz, 1H), 4.01 (s, 3H), 2.45 (s, 3H), 1.87-1.83 (m, 9H).

Example 10 Preparation of (2-((2-((4-(1-amino-2-methylpropan-2-yl)-2-methoxyphenyl)amino)-5-chloropyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide (Compound T-10)

(73) ##STR00053##

(74) The following route was used for the synthesis:

(75) ##STR00054## ##STR00055##

(76) Step 1 Synthesis of Compound 23

(77) To a 50 mL single-necked flask equipped with a magnetic stirrer compound 22 (2 g, 13.6 mmol) and THF (40 mL) were added, and the mixture was stirred to form a solution.

(78) After cooled to 0° C., potassium t-butoxide (4.7 g, 40.8 mmol) was added. After stirring for 10 minutes, iodomethane (5.8 g, 40.8 mmol) was added dropwise. After the addition was finished, the ice bath was removed and the reaction was stirred overnight at room temperature under a nitrogen atmosphere. The reaction was quenched by the addition of water (60 mL), and extracted with ethyl acetate (40 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was passed through a silica gel column to obtain 1.84 g of a white solid. Yield: 77.3%. LC-MS(APCI): m/z=176.2 (M+1).sup.+.

(79) Step 2 Synthesis of Compound 24

(80) To a 50 mL single-necked flask equipped with a magnetic stirrer tetrabutylammonium nitrate (6.7 g, 21.94 mmol), 18-crown-6 (845 mg, 3.2 mmol) and dichloromethane (30 mL) were added, and the mixture was stirred to form a solution. After cooled to 0° C., TFAA (13 mL) was added dropwise under a nitrogen atmosphere, and the solution was stirred for 15 minutes. A solution of compound 23 (4.0 g, 22.86 mmol) in dichloromethane (10 mL) was added dropwise, and after the addition, the ice bath was removed, and the reaction was stirred at room temperature for 2 hours, the reaction was quenched by water (30 mL) and saturated aqueous sodium bicarbonate solution (30 mL), and the solution was stirred for 5 minutes. The organic layer was separated, the aqueous phase was extracted with dichloromethane (40 mL×2), the organic phases were combined and dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was passed through a silica gel column to obtain 3.1 g of anhydrous oil. Yield: 61.6%. LC-MS(APCI): m/z=221.1 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ (ppm): 7.87 (d, J=8.8 Hz, 1H), 7.23 (d, J=2.0 Hz, 1H), 7.09 (dd, J=8.8 Hz, J=2.0 Hz, 1H), 4.00 (s, 3H), 1.76 (s, 6H).

(81) Step 3 Synthesis of Compound 25

(82) To a 100 mL single-necked flask equipped with a magnetic stirrer and a condenser tube compound 24 (3.2 g, 14.9 mmol) and ethanol/water (60 mL, 3/1) were added, and the mixture was stirred to form a solution. Reduced iron powder (70.5 mmol, 3.2 g) and ammonium chloride (14.9 mmol, 790 mg) were added, the reaction was heated to reflux under a nitrogen atmosphere and reacted at this temperature for 1 hour. The reaction was cooled to room temperature, and filtered, and the filter cake was washed with ethanol (10 mL). The resulting solution was concentrated to remove the organic solvent, and extracted with dichloromethane (50 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give 2.6 g of a brown solid. Yield: 91.8%. LC-MS(APCI): m/z=191.1 (M+1).sup.+.

(83) Step 4 Synthesis of compound 26

(84) To a 50 mL single-necked flask equipped with a magnetic stirrer and a condenser tube compound 25 (1.5 g, 7.9 mmol), compound 7 (2.5 g, 7.9 mmol) and ethylene glycol monomethyl ether (15 mL) were added, and the mixture was stirred to form a solution. A solution of hydrogen chloride in isopropanol (11.8 mmol, 2.4 mL, 5M) was added dropwise, and the temperature was raised to 120° C. under a nitrogen atmosphere and the reaction was stirred at this temperature overnight. The reaction was cooled to room temperature, water (30 mL) and saturated sodium bicarbonate (30 mL) were added, and the resulting solution was extracted with dichloromethane (40 mL×3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was passed through a silica gel column to give 1.2 g of a white solid. Yield: 32.4%.

(85) LC-MS(APCI): m/z=470.1 (M+1).sup.+.

(86) Step 5 Synthesis of Compound T-10

(87) To a 50 mL single-necked flask equipped with a magnetic stirrer and a condenser tube compound 26 (1.2 g, 2.56 mmol) and THF (10 mL) were added, and the mixture was stirred to form a solution. A borane tetrahydrofuran solution (8 mL, 8.0 mmol, 1M) was added dropwise, under nitrogen atmosphere, the reaction was heat to flux and was stirred at this temperature for 4 hours. The reaction was cooled to room temperature, the solvent was distilled off under reduced pressure, and the residue was passed through a silica gel column to give 460 mg of a white solid. Yield: 38.0%. LC-MS(APCI): m/z=474.2 (M+1).sup.+. .sup.1H NMR(400 MHz, CDCl.sub.3) δ (ppm): 10.82 (s, 1H), 8.59-8.55 (m, 1H), 8.29 (d, J=8.4 Hz, 1H), 8.08 (s, 1H), 7.58 (t, J=8.4 Hz, 1H), 7.50 (s, 1H), 7.33-7.30 (m, 1H), 7.17-7.14 (m, 1H), 6.88-6.85 (m, 2H), 3.91 (s, 3H), 3.05 (s, 2H), 1.85 (s, 3H), 1.82 (s, 3H), 1.45 (s, 6H).

Example 11 and Example 12 Preparation of (2-((5-chloro-2-((2-methoxy-4-(2-methyl-1-(methylamino)propan-2-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide (compound T-11) and (2-((5-chloro-2-((4-(1-(dimethylamino)-2-methylpropan-2-yl)-2-methoxyphenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide (Compound T-12)

(88) ##STR00056##

(89) The following route was used for the synthesis: IDC-51 C3

(90) ##STR00057##

(91) To a 50 mL single-necked flask equipped with a magnetic stirrer compound T-10 (300 mg, 0.63 mmol) and methanol (10 mL) were added, and the mixture was stirred to form a solution. Formaldehyde (15 mg, 0.5 mmol) and glacial acetic acid (1 drop) were added. The reaction was stirred for 10 minutes under a nitrogen atmosphere. After cooled to 0° C., NaBH.sub.3CN (40 mg, 0.63 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. The reaction was quenched by adding water (20 mL), and the solution was extracted with dichloromethane (20 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was passed through a silica gel column to give T-11, 50 mg of a white solid. Yield: 16.3%. LC-MS(APCI): m/z=488.2 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ (ppm): 10.87 (s, 1H), 8.61-8.58 (m, 1H), 8.40 (d, J=8.4 Hz, 1H), 8.14 (s, 1H), 7.59-7.55 (m, 2H), 7.35-7.30 (m, 1H), 7.22-7.17 (m, 1H), 7.03 (d, J=2.0 Hz, 1H), 6.93 (dd, J=8.0 Hz, J=2.0 Hz, 1H), 4.00 (s, 3H), 3.15 (s, 2H), 2.50 (s, 3H), 1.87 (s, 3H), 1.84 (s, 3H), 1.61 (s, 6H);

(92) The residue passing through the silica gel column also gave T-12, 150 mg of a white solid, Yield: 30.0%. LC-MS(APCI): m/z=502.2 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ (ppm): 10.87 (s, 1H), 8.61-8.58 (m, 1H), 8.40 (d, J=8.4 Hz, 1H), 8.14 (s, 1H), 7.59-7.55 (m, 2H), 7.35-7.30 (m, 1H), 7.22-7.17 (m, 1H), 7.03 (d, J=2.0 Hz, 1H), 6.93 (dd, J=8.0 Hz, J=2.0 Hz, 1H), 4.00 (s, 3H), 3.15 (s, 2H), 2.50 (s, 6H), 1.87 (s, 3H), 1.84 (s, 3H), 1.61 (s, 6H).

Example 13 Preparation of (2-((2-((4-(2-aminopropan-2-yl)-2-methoxyphenyl)amino)-5-chloropyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide (Compound T-13)

(93) ##STR00058##

(94) The following route was used for the synthesis:

(95) ##STR00059##

(96) Step 1 Synthesis of Compound 27

(97) To a 50 mL single-necked flask equipped with a magnetic stirrer compound 26 (1.6 g, 3.41 mmol) and water (10 mL) were added, and then concentrated sulfuric acid (10 mL) was added with stirring, and the reaction solution was heated to 100° C. under a nitrogen atmosphere, and was stirred at this temperature overnight. The reaction was cooled to room temperature, the pH thereof was adjusted to about 5 with aqueous NaOH (2M), and the resulting solution was extracted with the dichloromethane (40 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was passed through a silica gel column to obtain 850 mg of a white solid. Yield: 51.1%. LC-MS(APCI): m/z=487.2 (M−1).sup.−.

(98) Step 2 Synthesis of Compound T-13

(99) To a 50 mL single-necked flask equipped with a magnetic stirrer compound 27 (200 mg, 0.41 mmol) and methylene chloride (10 mL) were added, and the mixture was stirred to form a solution. Triethylamine (0.53 mL, 0.53 mmol) was added, and the reaction was stirred under a nitrogen atmosphere for 15 minutes before diphenyl azide phosphate (DPPA, 147 mg, 0.53 mmol) was added, and the reaction was further stirred at room temperature for 3 hours. The solvent was distilled off under reduced pressure, and the residue was dissolved in toluene (10 mL), and the resulted solution was heated to reflux for 2 hours. After cooled to room temperature, hydrochloric acid (20 mL, 6M) was added, and the reaction was refluxed again for 3 hours. The reaction was cooled to room temperature, pH thereof was adjusted to about 10 with aqueous ammonia, and the resulting solution was extracted with ethyl acetate (30 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was passed through a silica gel column to give 110 mg of a white solid. Yield: 58.4%. LC-MS(APCI): m/z=460.2 (M+1).sup.+. .sup.1H NMR (400 MHz, DMSO-D.sub.6) δ (ppm): 11.20 (s, 1H), 8.50-8.47 (m, 1H), 8.25-8.15 (m, 4H), 7.83 (d, J=8.8 Hz, 1H), 7.60-7.55 (m, 1H), 7.50 (t, J=8.0 Hz, 1H), 7.30 (s, 1H), 7.15 (t, J=8.0 Hz, 1H), 7.03 (dd, J=8.8 Hz. J=2.0 Hz, 1H), 3.87 (s, 3H), 1.79 (s, 3H), 1.75 (s, 3H), 1.63 (s, 6H).

Example 14 and Example 15 Preparation of (2-((5-chloro-2-((4-(2-(dimethylamino)propan-2-yl)-2-methoxyphenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide (Compound T-14) and (2-((5-chloro-2-((2-methoxy-4-(2-(methylamino)propan-2-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide (Compound T-15)

(100) ##STR00060##

(101) The following route was used for the synthesis:

(102) ##STR00061##

(103) To a 50 mL single-necked flask equipped with a magnetic stirrer compound T-13 (300 mg, 0.65 mmol) and methanol (10 mL) were added, and the mixture was stirred to form a solution. Formaldehyde (15 mg, 0.5 mmol) and glacial acetic acid (1 drop) were added. The reaction was stirred for 10 minutes under a nitrogen atmosphere. The reaction was cooled to 0° C., to which NaBH.sub.3CN (42 mg, 0.65 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. The reaction was quenched by adding water (20 mL), and extracted with dichloromethane (20 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was passed through a silica gel column to obtain T-14, 150 mg of a white solid. Yield: 30.0%. LC-MS(APCI): m/z=488.2 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ (ppm): 10.87 (s, 1H), 8.58-8.57 (m, 1H), 8.42 (d, J=8.4 Hz, 1H), 8.14 (s, 1H), 7.82 (br s, 1H), 7.61 (s, 1H), 7.56 (t, J=8.0 Hz, 1H), 7.31-7.28 (m, 1H), 7.17 (t, J=8.0 Hz, 1H), 6.98 (dd, J=8.4 Hz, J=2.0 Hz, 1H), 4.08 (s, 3H), 2.62 (s, 6H), 1.85 (s, 3H), 1.82 (s, 3H), 1.81 (s, 6H);

(104) The residue passing through the silica gel column also gave T-15, 50 mg of a white solid, Yield: 15.0%. LC-MS(APCI): m/z=474.2 (M+1).sup.+. .sup.1H NMR (400 MHz, DMSO-D.sub.6) δ (ppm): 11.20 (s, 1H), 9.53 (br s, 1H), 8.47-8.44 (m, 1H), 8.21 (s, 1H), 8.18 (s, 1H), 7.88 (d, J=8.4 Hz, 1H), 7.60-7.55 (m, 1H), 7.51 (t, J=8.0 Hz, 1H), 7.42 (s, 1H), 7.16 (t, J=8.0 Hz, 1H), 7.07 (dd, J=8.8 Hz. J=2.0 Hz, 1H), 3.89 (s, 3H), 2.22 (s, 3H), 1.79 (s, 3H), 1.75 (s, 3H), 1.69 (s, 6H).

Example 16 Preparation of (2-((2-((4-(1-aminopropan-2-yl)-2-methoxyphen yl)amino)-5-chloropyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide (Compound T-16)

(105) ##STR00062##

(106) The following route was used for the synthesis:

(107) ##STR00063## ##STR00064##

(108) Step 1 Synthesis of Compound 28

(109) To a 50 mL single-necked flask equipped with a magnetic stirrer compound 22 (4 g, 27.2 mmol) and THF (40 mL) were added, and the mixture was stirred to form a solution. After cooled to −10° C., LiHMDS (29.92 mL, 29.92 mmol, 1M) was added dropwise. The solution was stirred for 10 minutes, methyl iodide (4.3 g, 29.92 mmol) was added dropwise. After the addition, the reaction was stirred overnight at room temperature under a nitrogen atmosphere at 0° C. The reaction was quenched by adding water (60 mL), and extracted with ethyl acetate (40 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was passed through a silica gel column to obtain 3.2 g of a white solid. Yield: 73.5%. LC-MS(APCI): m/z=176.2 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl3) δ (ppm): 7.33-7.28 (m, 1H), 6.96-6.87 (m, 3H), 3.89 (q, J=7.2 Hz, 1H), 3.84 (s, 3H), 1.87 (d, J=7.2 Hz, 3H).

(110) Step 2 Synthesis of Compound 29

(111) To a 50 mL single-necked flask equipped with a magnetic stirrer tetrabutylammonium nitrate (6.7 g, 21.94 mmol), 18-crown-6 (845 mg, 3.2 mmol) and dichloromethane (30 mL) were added, and the mixture was stirred to form a solution. After cooled to 0° C., TFAA (13 mL) was added dropwise under a nitrogen atmosphere, and the solution was stirred for 15 minutes. A solution of compound 28 (3.7 g, 22.86 mmol) in dichloromethane (10 mL) was added dropwise, after the addition was finished, the ice bath was removed. The reaction was stirred at room temperature for 2 hours. The reaction was quenched with water (30 mL) and saturated aqueous sodium bicarbonate solution (30 mL), and the solution was stirred for 5 minutes. The organic layer was separated, and the aqueous phase was extracted with dichloromethane (40 mL×2). The organic phases were combined and dried over anhydrous sodium sulfate, filtered, and concentrated, the residue was passed through a silica gel column to obtain 3.1 g of anhydrous oil. Yield: 68.6%. LC-MS(APCI): m/z=207.1 (M+1).sup.+.

(112) Step 3 Synthesis of Compound 30

(113) To a 100 mL single-necked flask equipped with a magnetic stirrer and a condenser tube compound 29 (3.1 g, 14.9 mmol) and ethanol/water (60 mL, 3/1) were added, and the mixture was stirred to form a solution. Reduced iron powder (70.5 mmol, 3.2 g) and ammonium chloride (14.9 mmol, 790 mg) were added, the reaction was heated to reflux under a nitrogen atmosphere and reacted at this temperature for 1 hour. The reaction was then cooled to room temperature, filtered, the filter cake was washed with ethanol (10 mL), and organic solvent was removed by concentration. The resulting solution was extracted with dichloromethane (50 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give 2.4 g of a brown solid. Yield: 91.5%. LC-MS(APCI): m/z=177.1 (M+1).sup.+.

(114) Step 4 Synthesis of Compound 31

(115) To a 50 mL single-necked flask equipped with a magnetic stirrer and a condenser tube compound 30 (1.4 g, 7.9 mmol), compound 7 (2.5 g, 7.9 mmol) and ethylene glycol monomethyl ether (15 mL) were added, and the mixture was stirred to form a solution. A solution of hydrogen chloride in isopropanol (11.8 mmol, 2.4 mL, 5M) was added dropwise, and the temperature was raised to 120° C. under a nitrogen atmosphere and the reaction was stirred at this temperature overnight. The reaction was then cooled to room temperature, water (30 mL) and saturated sodium bicarbonate (30 mL) were added, and the resulting was extracted with dichloromethane (40 mL×3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was passed through a silica gel column to give 1.2 g of a white solid. Yield: 33.4%. LC-MS(APCI): m/z=456.1 (M+1).sup.+.

(116) Step 5 Synthesis of Compound T-16

(117) To a 50 mL single-necked flask equipped with a magnetic stirrer and a condenser tube compound 31 (1.2 g, 2.64 mmol) and THF (10 mL) were added, and the mixture was stirred to form a solution. Aborane tetrahydrofuran solution (8 mL, 8.0 mmol, 1M) was added dropwise, heated to reflux under nitrogen atmosphere and stirred at this temperature for 4 hours. The reaction was cooled to room temperature, the solvent was distilled off under reduced pressure, and the residue was passed through a silica gel column to obtain 460 mg of a white solid. Yield: 37.8%. LC-MS(APCI): m/z=460.2 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ (ppm): 10.77 (s, 1H), 8.55-8.52 (m, 1H), 8.19 (d, J=8.8 Hz, 1H), 8.02 (s, 1H), 7.53 (t, J=8.0 Hz, 1H), 7.42 (s, 1H), 7.29-7.25 (m, 1H), 7.16-7.14 (m, 1H), 6.79-6.77 (m, 2H), 3.89 (s, 3H), 3.18-3.15 (m, 2H), 3.05-3.02 (m, 1H), 1.84 (d, J=1.6 Hz, 3H), 1.80 (d, J=1.6 Hz, 3H), 1.36 (d, J=6.4 Hz, 3H).

Example 17 and Example 18 Preparation of (2-((5-chloro-2-((4-(1-(dimethylamino)propan-2-yl)-2-methoxyphenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide (Compound T-17) and (2-((5-chloro-2-((2-methoxy-4-(1-(methylamino)propan-2-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide (Compound T-18)

(118) ##STR00065##

(119) The following route was used for the synthesis:

(120) ##STR00066##

(121) To a 50 mL single-necked flask equipped with a magnetic stirrer compound T-16 (300 mg, 0.65 mmol) and methanol (10 mL) were added, and the mixture was stirred to form a solution. Formaldehyde (15 mg, 0.5 mmol) and glacial acetic acid (1 drop) were added. The reaction was stirred for 10 minutes under a nitrogen atmosphere. The reaction was then cooled to 0° C., NaBH.sub.3CN (42 mg, 0.65 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. The reaction was quenched by adding water (20 mL), and the resulting solution was extracted with dichloromethane (20 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was passed through a silica gel column to obtain T-17, 150 mg of a white solid. Yield: 30.0%. LC-MS(APCI): m/z=488.2 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ (ppm): 10.85 (s, 1H), 8.60-8.58 (m, 1H), 8.34 (d, J=8.4 Hz, 1H), 8.13 (s, 1H), 7.56 (t, J=8.0 Hz, 1H), 7.52 (s, 1H), 7.35-7.29 (m, 1H), 7.21-7.16 (m, 1H), 6.87 (s, 1H), 6.82-6.79 (m, 1H), 3.94 (s, 3H), 3.37-3.32 (m, 1H), 3.10-3.02 (m, 1H), 2.96-2.94 (m, 1H), 2.59 (s, 6H), 1.87 (s, 3H), 1.84 (s, 3H), 1.41 (d, J=6.4 Hz, 3H);

(122) The residue was passing through the silica gel column also gave T-18 as 50 mg of a white solid. Yield: 15.0%. LC-MS(APCI): m/z=474.2 (M+1).sup.+. 1H NMR (400 MHz, CDCl.sub.3) 5 (ppm): 10.85 (s, 1H), 8.60-8.58 (m, 1H), 8.34 (d, J=8.4 Hz, 1H), 8.13 (s, 1H), 7.56 (t, J=8.0 Hz, 1H), 7.52 (s, 1H), 7.35-7.29 (m, 1H), 7.21-7.16 (m, 1H), 6.87 (s, 1H), 6.82-6.79 (m, 1H), 3.94 (s, 3H), 3.37-3.32 (m, 1H), 3.10-3.02 (m, 1H), 2.96-2.94 (m, 1H), 2.59 (s, 3H), 1.87 (s, 3H), 1.84 (s, 3H), 1.41 (d, J=6.4 Hz, 3H).

Example 19 Preparation of (2-((2-((4-(2-aminoethyl)-2-methoxy-5-methylphenyl)amino)-5-chloropyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide (Compound T-19)

(123) ##STR00067##

(124) The following route was used for the synthesis:

(125) ##STR00068## ##STR00069##

(126) Step 1 Synthesis of Compound 34

(127) To a 50 mL single-necked flask equipped with a magnetic stirrer NaOH (2.7 g, 67.1 mmol) and DMSO (10 mL) were added, the solution was cooled to 0° C., and compound 32 (1.12 g, 6.71 mmol) and compound 33 (1.0 g, 6.71 mmol) in DMSO (10 mL) was added dropwise, and the reaction mixture was further stirred for 2 hours under nitrogen atmosphere. The reaction was quenched by adding water (50 mL), the resulting solution was extracted with ethyl acetate (40 mL×3), and washed with water (60 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was passed through a silica gel column as 1.3 g of a white solid. Yield: 94.2%. LC-MS(APCI): m/z=207.1 (M+1).sup.+.

(128) Step 2 Synthesis of compound 35

(129) To a 100 mL single-necked flask equipped with a magnetic stirrer and a condenser tube compound 34 (3.1 g, 14.9 mmol) and ethanol/water (60 mL, 3/1) were added, and the mixture was stirred to form a solution. Reduced iron powder (70.5 mmol, 3.2 g) and ammonium chloride (14.9 mmol, 790 mg) were added, the reaction was heated to reflux under a nitrogen atmosphere and reacted at this temperature for 1 hour. The reaction was then cooled to room temperature, filtered, and the filter cake was washed with ethanol (10 mL). The organic solvent was removed by concentration, and the resulting solution was extracted with dichloromethane (50 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give 2.4 g of a brown solid. Yield: 91.5%. LC-MS(APCI): m/z=177.1 (M+1).sup.+.

(130) Step 3 Synthesis of compound 36

(131) To a 50 mL single-necked flask equipped with a magnetic stirrer and a condenser tube compound 35 (1.4 g, 7.9 mmol), compound 7 (2.5 g, 7.9 mmol) and ethylene glycol monomethyl ether (15 mL) were added, and the mixture was stirred to form a solution. A solution of hydrogen chloride in isopropanol (11.8 mmol, 2.4 mL, 5M) was added dropwise, and the temperature was raised to 120° C. under a nitrogen atmosphere and the reaction was stirred at this temperature overnight. The reaction was then cooled to room temperature, water (30 mL) and saturated sodium bicarbonate (30 mL) were added, and the resulting solution was extracted with dichloromethane (40 mL×3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was passed through a silica gel column to give 1.2 g of a white solid. Yield: 33.4%. LC-MS(APCI): m/z=456.1 (M+1).sup.+.

(132) Step 4 Synthesis of compound T-19

(133) To a 50 mL single-necked flask equipped with a magnetic stirrer and a condenser tube compound 31 (1.2 g, 2.64 mmol) and THF (10 mL) were added, and the mixture was stirred to form a solution. Aborane tetrahydrofuran solution (8 mL, 8.0 mmol, 1M) was added dropwise, the reaction was heated to reflux under nitrogen atmosphere and the reaction was stirred and reacted at this temperature for 4 hours. The reaction was then cooled to room temperature, the solvent was distilled off under reduced pressure, and the residue was passed through a silica gel column to obtain 460 mg of a white solid. Yield: 37.8%. LC-MS(APCI): m/z=460.2 (M+1).sup.+. .sup.1H NMR (400 MHz, DMSO-D.sub.6) δ (ppm): 8.47-8.44 (m, 1H), 8.14 (s, 1H), 8.04 (s, 1H), 7.60-7.55 (m, 2H), 7.41 (t, J=8.0 Hz, 1H), 7.16-7.13 (m, 2H), 6.86 (s, 1H), 3.80 (s, 3H), 2.94 (t, J=8.4 Hz, 2H), 2.81 (t, J=8.4 Hz, 2H), 2.17 (s, 3H), 1.79 (s, 3H), 1.76 (s, 3H).

Example 20 and Example 21 Preparation of (2-((5-chloro-2-((4-(2-(dimethylamino)ethyl)-2-methoxy-5-methylphenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide (Compound T-20) and (2-((5-chloro-2-((2-methoxy-5-methyl-4-(2-(methylamino)ethyl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide (Compound T-21)

(134) ##STR00070##

(135) The following route was used for the synthesis:

(136) ##STR00071##

(137) To a 50 mL single-necked flask equipped with a magnetic stirrer compound T-19 (300 mg, 0.65 mmol) and methanol (10 mL) were added, and the mixture was stirred to form a solution. Formaldehyde (15 mg, 0.5 mmol) and glacial acetic acid (1 drop) were added. The reaction was stirred for 10 minutes under a nitrogen atmosphere. The reaction was cooled to 0° C., NaBH.sub.3CN (42 mg, 0.65 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. The reaction was quenched by adding water (20 mL), and the resulting solution was extracted with dichloromethane (20 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was passed through a silica gel column to obtain T-20, 150 mg of a white solid. Yield: 30.0%. LC-MS(APCI): m/z=488.2 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl3) δ (ppm): 10.79 (s, 1H), 8.59-8.56 (m, 1H), 8.09-8.08 (m, 2H), 7.48 (t, J=8.0 Hz, 1H), 7.43 (s, 1H), 7.30-7.27 (m, 1H), 7.14-7.10 (m, 1H), 6.68 (s, 1H), 6.82-6.79 (m, 1H), 3.84 (s, 3H), 2.98-2.94 (m, 2H), 2.80-2.73 (m, 2H), 2.58 (s, 6H), 2.20 (s, 3H), 1.83 (s, 3H), 1.81 (s, 3H);

(138) The residue was passing through the silica gel column also gave T-21 as 50 mg of a white solid. Yield: 15.0%. LC-MS(APCI): m/z=474.2 (M+1).sup.+. 10.76 (s, 1H), 8.59-8.56 (m, 1H), 8.09-8.08 (m, 2H), 7.48 (t, J=8.0 Hz, 1H), 7.43 (s, 1H), 7.30-7.27 (m, 1H), 7.14-7.10 (m, 1H), 6.68 (s, 1H), 6.82-6.79 (m, 1H), 3.84 (s, 3H), 2.98-2.94 (m, 2H), 2.80-2.73 (m, 2H), 2.55 (s, 3H), 2.20 (s, 3H), 1.83 (s, 3H), 1.81 (s, 3H).

Example 22 Biological Evaluation of Compounds

(139) The compounds of the present disclosure are evaluated in multiple assays to determine their biological activity. For example, the compounds of the present disclosure may be tested for their ability to inhibit various protein kinases of interest. Some tested compounds showed potent inhibitory activity against ALK kinase.

(140) (1) Evaluation of Kinase Inhibition

(141) Compound preparation: The test compounds were dissolved in DMSO to make 20 mM stock solutions. The compounds were diluted in DMSO to 0.1 mM (a dilution with 100 times the final concentration) before use, and a 3-fold series gradient dilution with 11 concentrations was made. Dilutions with 4 times of the final concentration were prepared by diluting with buffer when dosing.

(142) Kinase assay: after preparing the buffer solution, the enzyme was mixed with pre-diluted compounds with different concentrations, and placed at room temperature for 30 minutes in duplicate. The corresponding substrate and ATP were added thereto and reacted at room temperature for 60 minutes (both a negative and a positive control were set). After the reaction, antibody was added for detection, and after incubation at room temperature for 60 minutes, Evnvision detection was carried out, and data was collected. Data was analyzed and fit according to XLfit5 software. IC.sub.50 was calculated by the following formula: (IC.sub.50=[(ABS test−ABS begin)/(ABS control−ABS begin)]×100). Wherein A represents IC.sub.50≤2 nM, B represents IC.sub.50 of 2-5 nM, C represents IC.sub.50 of 5-10 nM.

(143) The compounds of the present disclosure were tested in the above kinase inhibition assay, and were found to have potent activity against ALK and ALK[L1196M]. The results of the representative example compounds are summarized in Table 1 below.

(144) TABLE-US-00001 TABLE 1 IC.sub.50(nM) Example ALK No. ALK [WT] [L1196M]  1 C C  2 A B  3 A B  4 A B  5 A A  6 B B  7 B B  8 B B  9 B B 10 B B 11 B C 12 B C 13 A A 14 A B 15 A A 16 C C 17 C C 18 19 20 B B 21 B C
(2) Cytotoxicity Test

(145) The in vitro anti-proliferative activity of the compounds of the present disclosure against three types of cells that are cultured in vitro was tested by CellTiter-Glo method. The experimental results show that the compounds of the present disclosure have potent inhibition on the in vitro proliferation of EML4-ALK and EML4-ALK L1196M mutant cells that cultured in vitro.

(146) Cell lines: BaF3 parental; BaF3 [EML4-ALK] (From WuXi PharmaTech) and BaF3 [EMF4-AFK F1196M] (From WuXi PharmaTech); wherein, BaF3 parental was cultured in RPMI1640 medium containing 10 ng/ml IL-3, 10% fetal bovine serum, 100 U/ml penicillin, and 100 μg/ml streptomycin, BaF3 [EMF4-AFK] and BaF3 [EMF4-AFK F1196M] were cultured in RPMI1640 medium containing 10% fetal bovine serum, 100 U/ml penicillin, and 100 μg/ml streptomycin.

(147) Reagents and materials: RPMI-1640 (GIBCO, Cat. No. A10491-01); fetal bovine serum (GIBCO, Cat. No. 10099141); 0.25% trypsin-EDTA (GIBCO, Cat. No. 25200); penicillin-streptomycin, liquid (GIBCO, Cat. No. 15140-122); DMSO (Sigma, Cat. No. D2650); CellTiter-Glo Test Kit (Promega, Cat. No. G7572), and 96-well plate (Corning, Cat. No. 3365).

(148) Specific Experimental Procedures:

(149) 1. The test compounds were dissolved in DMSO to make stock solutions and subjected to a series gradient dilution to give solutions with 10-fold working concentration.

(150) 2. The cells in the logarithmic growth phase were diluted with the culture solution to adjust to the specific cell concentration, 90 μi of cell suspension was added to the 96-well plate to make the cell density reach the specified concentration. The plate was incubated in a incubator at 37° C., containing 5% carbon dioxide gas overnight.

(151) 3. 10 μl drug solutions were added to each well of the 96-well plate inoculated with cells. 10 concentrations of the compounds were tested starting from the highest concentration of 20 μM with a 3-fold series gradient dilution in duplicate.

(152) 4. After continuously cultured for 72 hours, the cells were detected by CellTiter-Glo for the cell viability. The dose-effect curve was made using GraphPad Prism software and IC.sub.50 was calculated.

(153) The compounds of the present disclosure were tested in the above-mentioned cytotoxicity assay, finding out that the compounds of the present disclosure have potent activity against Ba/F3 AFK and Ba/F3 AFK[F1196M], and superior selectivity over Ba/F3 parental cells. The results of the inhibition effects on the proliferation of cancer cells in vitro by the representative examples compounds of the present disclosure were summarized in Table 2 below, wherein, A represents IC.sub.50<25 nM, B represents IC.sub.50 of 25-50 nM, C represents IC.sub.50 of 50-100 nM, D represents IC.sub.50 of 100-200 nM, E represents IC.sub.50 of 200-1000 nM, and F represents IC.sub.50≥1000 nM.

(154) TABLE-US-00002 TABLE 2 IC.sub.50(nM) Selectivity Example parental ALK ALK[L1196M] parental/ parental/ No. (Ba/F3) (Ba/F3) (Ba/F3) ALK ALK[L1196M]  1 F C D <50 <50  2 F D E <50 <50  3 F B D  50 to 100 <50  4 F A C 200 to 500 <50  5 F A B 200 to 500 100 to 200  6 F A D 200 to 500 <50  7 F C E <50 <50  8 F B D 100 to 200 <50  9 F C E  50 to 100 <50 10 F B D 100 to 200 <50 11 F B C 100 to 200  50 to 100 12 F B C 100 to 200 <50 13 F B D <50 <50 14 F A C 100 to 200 <50 15 F C D <50 <50 16 F E E  50 to 100 <50 17 F B D 100 to 200  50 to 100 20 F B B  50 to 100  50 to 100 21 F D E <50 <50
(3) Metabolic Stability Evaluation

(155) Experiments in microsomes: Human liver microsomes: 0.5 mg/mL, Xenotech; Rat liver microsomes: 0.5 mg/mL, Xenotech; Coenzyme (NADPH/NADH): 1 mM, Sigma Life Science; Magnesium chloride: 5 mM, 100 mM phosphate buffer (pH 7.4).

(156) Preparation of stock solution: Powder of the example compounds was accurately weighed and dissolved in DMSO to 5 mM. Preparation of phosphate buffer (100 mM, pH7.4): A pre-formulated 0.5M potassium dihydrogen phosphate (150 mL) was mixed with 0.5M dibasic potassium phosphate (700 mL). The pH of the mixture was adjusted to 7.4 with 0.5M dibasic potassium phosphate solution. The mixture was diluted 5-fold with ultrapure water before use, and magnesium chloride was added to obtain a phosphate buffer (100 mM) containing 100 mM potassium phosphate, 3.3 mM magnesium chloride, pH7.4. A NADPH regeneration system solution (containing 6.5 mM NADP, 16.5 mM G-6-P, 3 U/mL G-6-PD, 3.3 mM magnesium chloride) was prepared and placed on wet ice prior to use. Preparation of stop solution: acetonitrile solution containing 50 ng/mL propranolol hydrochloride and 200 ng/mL tolbutamide (internal standard). 25057.5 μL of phosphate buffer (pH 7.4) was taken into a 50 mL centrifuge tube, to which 812.5 μL human liver microsomes were added, and mixed to obtain a liver microsome dilution solution with a protein concentration of 0.625 mg/mL. 25057.5 μL of phosphate buffer (pH 7.4) was taken into a 50 mL centrifuge tube, to which 812.5 μL SD rat liver microsomes were added, and mixed to obtain a liver microsome dilution solution with a protein concentration of 0.625 mg/mL.

(157) Incubation of the samples: The stock solution of the respective compound was respectively diluted to 0.25 mM with an aqueous solution containing 70% acetonitrile, and used as a working solution, ready for use. 398 μL of the dilution solution of human liver microsomes or rat liver microsome were added to a 96-well incubation plate (N=2), respectively, and 2 μL of 0.25 mM working solution was added and mixed.

(158) Metabolic stability assay: 300 μL of pre-chilled stop solution was added to each well of a 96-well deep well plate and placed on ice as a stop plate. The 96 well incubation plate and NADPH regeneration system were placed in a 37° C. water bath box, shaken at 100 rpm and pre-incubated for 5 min. 80 μL of incubation solution was taken out from each well of the incubation plate and added to the stop plate, mixed, and replenished with 20 μL of NADPH regeneration system solution as a 0-min sample. 80 μL of NADPH regeneration system solution was added to each well of the incubation plate to start the reaction and start counting. The corresponding compound had a reaction concentration of 1 μM and the protein concentration was 0.5 mg/mL. Separately, 100 μL of the reaction solution was taken at 10, 30, and 90 min reaction, respectively, added to a stop plate, and vortexed for 3 minutes to terminate the reaction. The stop plate was centrifuged at 5000×g at 4° C. for 4 min. 100 μL of the supernatant was added to a 96-well plate to which 100 μL of distilled water was previously added, mixed, and analyzed by LC-MS/MS.

(159) Data analysis: The peak areas of the corresponding compound and internal standard were detected by LC-MS/MS system, and the ratio of the peak area of the compound to the internal standard was calculated. The slope was measured by plotting the natural logarithm of the percent of compound remaining versus time, and t.sub.1/2 and CL.sub.int were calculated according to the formula, where V/M is equal to 1/protein concentration.

(160) $t_{1/2}=-\frac{0.693}{\mathrm{slope}},{\mathrm{CL}}_{\mathrm{int}}=\frac{0.693}{t_{1/2}}•\frac{V}{M},t_{1/2}\left(\min \right);{\mathrm{CL}}_{\mathrm{int}}\left(\mathrm{.Math.L}\text{/}\min \text{/}\mathrm{mg}\right).$

(161) The compounds of the present disclosure were evaluated for their metabolic stability in human and rat liver microsomes. The half-life and the intrinsic liver clearance rate as indicators of metabolic stability are shown in Table 3 below. The compound of the present disclosure can significantly improve the metabolic stability.

(162) TABLE-US-00003 TABLE 3 Human liver microsome Rat liver microsome experiment experiment Example CL.sub.int CL.sub.int No. t.sub.1/2 (min) (uL/min/mg) t.sub.1/2 (min) (uL/min/mg)  1 131.2 10.6 31.9 43.4  2 159.6 8.7 164.6 8.4  3 90.1 15.4 27.5 50.4  4 51.9 26.7 39.9 34.7  5 80.0 17.3 38.6 35.9  6 47.6 29.1 16.3 85.1  7 243.9 5.7 340.3 4.1  8 56.3 24.6 48.2 28.8  9 123.1 11.3 139.6 9.9 10 55.0 25.2 92.0 15.1 11 58.2 23.8 193.7 7.2 12 18.9 73.5 16.0 86.8 13 118.8 11.7 152.3 9.1 14 46.4 29.9 41.5 33.4 15 66.9 20.7 78.8 17.6 16 64.6 21.5 118.4 11.7 17 74.3 18.6 46.7 29.7 20 258.8 5.4 42.9 32.3 21 50.0 27.7 192.7 7.2
(4) Pharmacokinetic Experiment in Rats

(163) 6 male Sprague-Dawley rats (7-8 weeks old, and weighing approximately 210 g) were divided into 2 groups with 3 rats in each group. The rats were intravenously or orally administered a single dose of compound (10 mg/kg orally) to compare pharmacokinetic differences.

(164) The rats were raised on standard food and water. Fasting was started 16 hours before the test. The drug was dissolved with PEG400 and dimethyl sulfoxide. The blood samples were collected from eyelids at the time points of 0.083 hours, 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 12 hours, and 24 hours after administration.

(165) Rats were briefly anesthetized after inhalation of diethyl ether and 300 μL of blood sample was collected from the eyelids into test tubes. There was 30 μL of 1% heparin salt solution in the test tube. Tubes were dried at 60° C. overnight before use. After the blood sample was collected at the last time point, the rats were sacrificed after ether anesthesia.

(166) Immediately after the collection of the blood sample, the test tube was gently inverted at least 5 times to ensure sufficient mixing and then placed on ice. The blood sample was centrifuged at 5000 rpm at 4° C. for 5 minutes to separate the plasma from the red blood cells. 100 μL of plasma was aspirated into a clean plastic centrifuge tube with a pipette, marking with the name of the compound and time point. Plasma was stored at −80° C. prior to analysis. The concentration of the compound of the present disclosure in plasma was determined by LC-MS/MS. The pharmacokinetic parameters were calculated based on the blood concentration of each animal at different time points.

(167) Experiments showed that the compounds of the present disclosure have better pharmacokinetic properties in animals, and therefore have better pharmacodynamics and treatment effects.

(168) The above is a further detailed description of the present disclosure in conjunction with specific embodiments, and it cannot be assumed that the specific implementation of the present disclosure is limited to these descriptions. For ordinary artisan in the technical field to which the present disclosure belongs, without deviating from the concept of the present disclosure, various simple deductions or replacements may be made, which should be regarded as falling within the protection scope of the present disclosure.